Cytotoxic and anti-mitotic compounds, and methods of using the same.

The invention relates to biologically active compounds, compositions comprising the same, and methods of using such biologically active compounds and compositions for the treatment of cancer and other diseases.

Description of the related technique

Talpir, R. et al. (1994) Tetrahedron Lett. 35: 4453-6, describe the natural hemiasterlin compound, a stable tripeptide obtained from marine sponges that causes depolymerization of microtubules and mitotic arrest in cells. Hemiasterlin consists of unusual and highly congested amino acids, characteristics that are thought to contribute to its activity. A number of groups have modified particular structural elements of hemiasterlin to assess the structure-activity relationships and evaluate the activity of hemiasterline analogs. See, for example, Zask et al., Bioorganic & Medicinal Chemistry Letters, 14: 4353-4358, 2004; Zask et al., J Med Chem, 47: 4774-4786, 2004; Yamashita et al,

Bioorganic & Medicinal Chemistry Letters, 14: 5317-5322,

2004; PCT / GB96 / 00942; WO 2004/026293; W096 / 33211; and US

7,579,323.

Hemiasterlin analogues with modifications in the A-segment, or the amino terminal segment, have been described (see, for example, Zask et al., J Med Chem, 47: 4774-4786, 2004; Yamashita et al., Bioorganic & Medicinal Chemistry Letters, 14: 5317-5322, 2004; US 7,579,323). US 7,579,323 describes a hemiasterlin analog, referred to as HTI-286, in which the indole residue is substituted by a phenyl group. HTI-286 exhibits potent antimitotic activity and has been evaluated in clinical trials for the treatment of cancer (Ratain et al., Proc Am Soc Clin Oncol, 22: 129, 2003).

Hemiasterlin analogs with modifications in the D-segment, or the carboxy terminal segment, have also been reported (see, for example, WO 2004/026293;

Zask et al., Bioorganic & Medicinal Chemistry Letters, 14: 4353-4358, 2004; Zask et al., J Med Chem, 47: 4774-4786, 2004). Most modifications to the carboxy terminal result in compounds with substantially decreased potency compared to parental carboxylic acids. See, for example, WO 2004/026293, in particular Table 12. Zask et al., (J Med Chem, 47: 4774-4786, 2004) also reported that amide analogs prepared using simple cyclic and acyclic amines exhibit significantly reduced potency (reductions of one to three orders of magnitude). Among the few tolerated modifications, Zask et al., (Bioorganic & Medicinal Chemistry Letters, 14: 4353-4358, 2004) report that the addition of esterified cyclic amino acids at the carboxy terminal end produces tetrapeptide analogs with terminal ends containing type esters prolific, some of which exhibit a potency comparable to the parental compound in a proven cancer cell line.

The potent cytotoxic and antimitotic compositions are highly desired for the treatment of a number of devastating disorders, including cancer. While a wide variety of hemiasterlin analogs have been generated, many, including a wide variety of compounds with modifications at the carboxy terminal end, exhibit reduced potency that limits their usefulness in medical treatment methods.

For the reasons stated, while progress has been made in this field, there is a need for additional potent antimitotic and cytotoxic compounds that have preferred characteristics that make them suitable for the treatment of a variety of disorders, including cancer. The present disclosure satisfies these needs and provides other related advantages.

SUMMARY OF THE INVENTION

In summary, the present description refers to biologically active compounds, compositions comprising the same, and methods of use of such compounds and compositions.

In one embodiment, compounds having the following structure (I) are provided:

<img file="MX368258B_D0001.tif" />

'2 R!

(I) where:

Ri and R₂ they are independently selected from the group consisting of: H and a saturated or unsaturated residue having a linear, branched or non-aromatic cyclic skeleton containing from one to ten carbon atoms, and the carbon atoms are optionally substituted with: -OH, -I, -Br, Cl, -F, -CN, -CO2H, -CHO, -COSH, or -NO2; or R2 and R5 fuse and form a ring;

R3 and R₄ are independently selected from the group consisting of: H, R, ArR-, or R₃ and R₄ join together to form a ring;

R5 is selected from the group consisting of: H, R, ArR- and Ar;

or R₅ and R₂ they fuse and form a ring;

Rg is selected from the group consisting of: H, R, and ArR-;

R7 and Rg are independently selected from the group consisting of: H, R, and ArR-; Y

R9 is:

OO

II H II wxry — C — N — S — R₁₄

II

Or where,

R is defined as a saturated or unsaturated residue having a linear, branched, or non-aromatic cyclic skeleton containing from one to ten carbon atoms, from zero to four nitrogen atoms, from zero to four oxygen atoms, and from zero at four sulfur atoms, and the carbon atoms are optionally substituted with: = 0, = S, OH, -OR10, -0₂CRio, -SH,

-Srio, -SOCRio, -NH₂, -NHRio, -N (Rio)₂, -NHCORio, -NRiqCORiq, I, -Br, -Cl, -F, -CN, -CO₂H, -C0₂Rio, -CHO, -COR₁₀, -CONH₂, CONHRio, -CON (Rio) 2, -COSH, -COSRio, -NO₂, -SO₃H, -SOR₁₀, S0₂Rior where Ri₀ it is a linear, branched or cyclic, saturated or unsaturated alkyl group of one to ten carbons;

the ring formed by the union of R₃ and R₄ it is a non-aromatic cyclic skeleton of three to seven members within the definition of R,

Y is defined as a residue selected from the group consisting of: a linear, saturated or unsaturated alkyl group of one to six carbons, optionally substituted with R, ArR-, or X; Y,

X is defined as a residue selected from the group consisting of: -OH, -OR, = 0, = S, -O₂CR, -SH, -SR, -SOCR, —NH₂, NHR, -N (R)₂, -NHCOR, -NRCOR, -I, -Br, -Cl, -F, -CN, -CO₂H, CO₂R, -CHO, -COR, -CONH₂, -CONHR, -CON (R)₂, -COSH, -COSR, NO₂—SO₃H, -SOR, and —SO₂R;

R₁₄ is selected from the group consisting of optionally substituted alkyl, optionally substituted alkylamino, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heterocyclyl and optionally substituted heteroaryls, COR₂₄, CSR₂₄, -OR₂₄, and -NHR₂₄, where each R₂₄ it is, independently, alkyl optionally substituted with halogen, -OH or -SH;

or a stereoisomer, prodrug or pharmaceutically acceptable salt thereof.

In one embodiment, Ar is an aromatic ring selected from the group consisting of: phenyl, naphthyl, anthracil, pyrrolyl.

In one embodiment, compounds having the following structure (the) are provided where:

R14

<img file="MX368258B_D0002.tif" />

is selected from the group consisting of optionally substituted alkyl, optionally substituted alkylamino, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heterocyclyl, optionally substituted heteroaryl, COR₂4, CSR₂4, -OR₂4, and -NHR₂₄, where each R₂₄ it is, independently, alkyl optionally substituted with halogen, -OH or -SH;

R₁₅ it is selected from the group consisting of optionally substituted alkyl, optionally substituted alkylamino, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heterocyclyl and optionally substituted heteroaryl;

R16 is selected from the group consisting of H and Ci_ 6 alkyl /

Ri7 is selected from the group consisting of H and C 1_z 6-acyl *

Laugh and R30 are independently selected from the group consisting of H, C1-6 alkyl and ~ SH, with the proviso that Laugh and R30 cannot be both H;

R19, ​​^ 20, R21 and R22 are independently H and C1-6 alkyl, at least one of R19 and R₂₀ it's H; or R20 and R21 form a double bond, R19 is H, and R₂₂ is H or Ci_ alkyl₆; Y

R23 is selected from the group consisting of H and Ci_ alkyl ₆; or a stereoisomer, prodrug or pharmaceutically acceptable salt thereof.

In a further embodiment, each optionally substituted alkyl, optionally substituted alkylamino, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heterocyclic and optionally substituted heteroaryl is, independently, optionally substituted with = 0, = S, -OH, -OR₂₄, -O₂CR₂₄, -SH, -sr₂₄, -socr₂₄, -nh₂, -n₃, -nhr₂₄, -n (r₂₄)₂, -nhcor₂₄, NR₂₄COR₂₄, -I, -Br, -Cl, -F, -CN, -CO₂H, -CO₂R₂₄, -CHO, -COR₂₄,

-CONH₂, -CONHR₂₄, -CON (R₂₄)₂, -cosh, -cosr₂₄, -do not₂-so₃h, SOR₂₄ or -SO2R24 θη where each R₂₄ it is, independently, alkyl optionally substituted with halogen, -OH or -SH.

In another additional embodiment, each optionally substituted aryl and optionally substituted heteroaryl is independently selected from the group consisting of optionally substituted phenyl, optionally substituted naphthyl, optionally substituted anthracil, optionally substituted phenanthryl, optionally substituted furyl, optionally substituted pyrrolyl, thiophenyl optionally substituted, optionally substituted benzofuryl, optionally substituted benzothiophenyl, optionally substituted quinolinyl, optionally substituted isoquinolinyl, optionally substituted imidazolyl, optionally substituted thiazolyl, optionally substituted oxazolyl, and optionally substituted pyridinyl.

In another additional embodiment, R15 is selected from one of the following structures (II), (III), (IV), (V):

<img file="MX368258B_D0003.tif" />

<img file="MX368258B_D0004.tif" />

(II)

<img file="MX368258B_D0005.tif" />

(III)

<img file="MX368258B_D0006.tif" />

item S · ,.

(IV) and

<img file="MX368258B_D0007.tif" />

(V) where:

Q is CR25 or N;

Z is C (R₂₅)₂, NR₂₅, S, or O, each R25 is independently selected from the group consisting of H, -OH, -R24, -OR₂₄, -0₂CR₂₄, -SH, -SR₂₄, SOCR₂₄, -NH₂, -N₃, -NHR₂₄, -N (R₂₄)₂, -NHCOR24, -NR24COR24, -R24NH2, -I, -Br, -Cl, -F, -CN, -CO₂H, -CO₂R₂₄, -CHO, -COR₂₄, -CONH₂, CONHR24, -CON (R₂₄) 2, -COSH, -COSR24, -NO2, -SO3H, -sor₂₄ bear₂R₂₄, θη where each R₂₄ it is, independently, alkyl optionally substituted with halogen, -OH or -SH.

In another additional mode, R15 is selected from the group consisting of:

<img file="MX368258B_D0008.tif" />

<img file="MX368258B_D0009.tif" />

wherein each R25 is independently selected from the group consisting of H, -OH, -R₂₄, -OR₂₄, -O₂CR₂₄, -SH, -SR₂4, -SOCR₂₄, -nh₂, -n₃, -NHR₂₄, -N (R₂₄)₂, -nhcor₂₄, -nr₂₄cor₂₄, R24NH2, -I, -Br, -Cl, -F, -CN, -CO₂H, -CO₂R₂₄, -CHO, -COR₂₄, ~ conh₂, -conhr₂₄, -CON (R₂₄)₂, -cosh, -cosr₂₄, -do not₂-so₃h, -SOR₂₄ bear₂R₂₄, where each R₂₄ it is, independently, alkyl optionally substituted with halogen, -OH or -SH.

In another additional mode, R15 is selected from the group consisting of:

<img file="MX368258B_D0010.tif" />

<img file="MX368258B_D0011.tif" />

<img file="MX368258B_D0012.tif" />

In another additional mode, R₁₅ it is:

<img file="MX368258B_D0013.tif" />

In another additional mode, R_i6, R17, Laugh, and R30 are each methyl.

In another additional mode, R₁₆, is H, R₁₇ It's methyl, Ri₈ it is methyl and R₃₀ It is methyl.

It is understood that any modality of the compounds of structure (la), as set forth above, and any specific substituent set forth herein for a group R₁₄, R15, Ri₆, R17, Ris, R19, R20 and R30 θη the compounds of structure (la), as set forth above, can be combined independently with other modalities and / or substituents of compounds of structure (I) to form modalities of This specification not specifically set forth above. In addition, in the event that a list of substituents is listed for any R_i4, R_i5, R_i6, R17, Ri₈, R₁₉, R₂o and R30 particular in a particular embodiment and / or claim, it is understood that each individual substituent can be removed from the particular modality and / or claim, and that the remaining list of substituents will be deemed to be within the scope of the present description.

In one embodiment, compounds that have the following structure (Ib) are provided:

<img file="MX368258B_D0014.tif" />

where:

R₂6 it is selected from the group consisting of optionally substituted alkyl, optionally substituted alkylamino, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heterocyclyl and optionally substituted heteroaryl;

R₂7 it is selected from the group consisting of optionally substituted alkyl, optionally substituted alkylamino, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heterocyclyl and optionally substituted heteroaryl;

Ri6 is selected from the group consisting of H and C6 alkyl /

Ri7 is selected from the group consisting of H and Ci_e / Y alkyl

Laugh is selected from the group consisting of Ci-β alkyl and

SH, or a stereoisomer, prodrug or pharmaceutically acceptable salt thereof.

In a further embodiment, each optionally substituted alkyl, optionally substituted alkylamino, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heterocyclyl and optionally substituted heteroaryl is, independently, optionally substituted with = 0, = S, -OH, -OR₂₈, -O₂CR₂8 &lt;-SH, -SR₂₈, -SOCR₂₈, -NH₂, -N₃, -NHR₂₈, -N (R₂₈)₂, -NHCOR28, -NR₂₈COR₂8, -I, -Br, -Cl, -F, -CN, -CO₂H, -CO₂R₂8, -CHO, -COR₂₈, -CONH₂, CONHR₂₈, -CON (R₂₈)₂, -COSH, -cosr₂₈, -do not₂-so₃h, -SOR₂₈ bear₂R₂₈, where each R₂8 it is, independently, alkyl optionally substituted with halogen, -OH or -SH.

In another additional embodiment, each optionally substituted aryl and optionally substituted heteroaryl is independently selected from the group consisting of optionally substituted phenyl, optionally substituted naphthyl, optionally substituted anthracyl, optionally substituted phenanthryl, optionally substituted furyl, optionally substituted pyrrolyl, optionally substituted thiophenyl , optionally substituted benzofuryl, optionally substituted benzothiophenyl, optionally substituted quinolinyl, optionally substituted isoquinolinyl, optionally substituted imidazolyl, optionally substituted thiazolyl, optionally substituted oxazolyl, and optionally substituted pyridinyl.

In another additional embodiment, R27 is selected from one of the following structures (II), (III), (IV), (V):

<img file="MX368258B_D0015.tif" />

(II)

<img file="MX368258B_D0016.tif" />

(III)

<img file="MX368258B_D0017.tif" />

(IV)

<img file="MX368258B_D0018.tif" />

(V) where:

Q is CR29 or N;

Z is C (R29) 2, NR29, S, or 0_; each R29 is independently selected from the group consisting of H, -OH, -OR₂₈, -O₂CR₂8, -SH, -SR₂₈, -SOCR₂₈, -NH₂,

N₃, -NHR₂₈, -N (R₂₈)₂, -NHCOR₂₈, -NR₂₈COR₂₈, -I, -Br, -Cl, -F, CN, -co₂h, -co₂r₂₈, -cho, -cor₂₈-conh₂, -conhr₂₈, -CON (R₂₈)₂, COSE, -COSR₂8, -NO₂, -SO3H, -SOR28 or - SO₂R28a where each R₂₈ it is, independently, alkyl optionally substituted with halogen, -OH or -SH.

In another additional mode, R₂7 It is selected from the group consisting of:

<img file="MX368258B_D0019.tif" />

<img file="MX368258B_D0020.tif" />

<img file="MX368258B_D0021.tif" />

<img file="MX368258B_D0022.tif" />

<img file="MX368258B_D0023.tif" />

In another additional mode, R₂7 it is:

In another additional mode, R₁₅, R₁₇ and R₁₈ They are each methyl.

In another additional mode, R₁₆ it's H, R₁₇ is methyl, and R₁₈ It is methyl.

It is understood that any modality of the compounds of structure (Ib), as set forth above, and any specific substituent set forth herein for a group R25, R26 / Laugh, R17, Ris, Laugh and R20 in the compounds of structure ( Ib), as stated above, they can be combined independently with other modalities and / or substituents of compounds of structure (I) to form modalities of the present specification not specifically set forth above. In addition, in the event that a list of substituents is listed for any R₂₅, R₂6, Ri6z Ri7z Laugh, Laugh and R20 in a particular embodiment and / or claim, it is understood that each individual substituent may be removed from the particular modality and / or claim, and that the remaining list of substituents will be considered to be within the scope of this description.

In one embodiment, the invention provides a method of manufacturing a compound having structure (I), (la) or (Ib).

In another embodiment, a pharmaceutical composition is provided comprising a compound having the structure (I), (la) or (Ib), or a pharmaceutically acceptable stereoisomer, salt or prodrug thereof, and an acceptable pharmaceutical carrier, diluent or excipient .

In another embodiment, a method is provided for using a compound having structure (I), (la) or (Ib), or a pharmaceutically acceptable stereoisomer, salt or prodrug thereof. In particular, the present description provides a method for the treatment of cancer in a mammal which comprises administering to an mammal in need thereof an effective amount of a compound having the structure (I), (la) or (Ib), or a stereoisomer, pharmaceutically acceptable salt or prodrug thereof, or a pharmaceutical composition comprising a compound having the structure (I), (la) or (Ib), or a stereoisomer, pharmaceutically acceptable salt or prodrug thereof, and a pharmaceutically acceptable carrier, diluent or excipient.

In another embodiment, the present description provides a method for inhibiting tumor growth in a mammal that comprises administering to an mammal in need thereof an effective amount of an emptiness having structure (I), (la) or (Ib), or a stereoisomer, salt or pharmaceutically acceptable prodrug thereof, or a pharmaceutical composition comprising a compound having the structure (I), (la) or (Ib), or a stereoisomer, pharmaceutically acceptable salt or prodrug thereof, and a pharmaceutically acceptable carrier, diluent or excipient.

In another embodiment, the present disclosure provides a method of killing cancer cells in vitro using a compound having the structure (I), (la) or (Ib), or a pharmaceutically acceptable stereoisomer, salt or prodrug thereof. In another embodiment, the present description provides a method for killing cancer cells in vivo in a mammal, which comprises administering to an mammal in need thereof an effective amount of a compound having structure (I), (la) or (Ib ), or a pharmaceutically acceptable stereoisomer, salt or prodrug thereof, or a pharmaceutical composition comprising a compound having the structure (I), (la) or (Ib), or a stereoisomer, pharmaceutically acceptable salt or prodrug thereof, and a pharmaceutically acceptable carrier, diluent or excipient.

In another embodiment, the present description provides a method for increasing the survival time of a mammal having cancer, which comprises administering to said mammal an effective amount of a compound having structure (I), (la) or (Ib) , or a pharmaceutically acceptable stereoisomer, salt or prodrug thereof, or a pharmaceutical composition comprising a compound having the structure (I), (la) or (Ib), or a stereoisomer, pharmaceutically acceptable salt or prodrug thereof, and a pharmaceutically acceptable carrier, diluent or excipient.

In one embodiment, compositions are provided comprising biologically active compounds having structure (I), (la) or (Ib), or a pharmaceutically acceptable stereoisomer, salt or prodrug thereof, directly or indirectly linked to a target residue.

In one embodiment, the invention provides compositions having the following structure:

(T) - (L) - (D) (VI) where (T) is a target residue, (L) is an optional linker, and (D) is a compound having structure (I), (la) or (Ib), or a pharmaceutically acceptable stereoisomer, salt or prodrug thereof. (D) covalently binds to (L), if (L) is present, or (T), if (L) is not present.

In a particular embodiment, (D) is a compound that has the structure (Ib).

In one embodiment, the target residue is an antibody. Accordingly, in one embodiment, antibody-drug conjugates (ADCs) are provided comprising compounds having the structure (I), (la) or (Ib), or a pharmaceutically acceptable stereoisomer, salt or prodrug thereof.

In one embodiment, the invention provides a method of manufacturing a composition having the structure (VI).

<td colspan="2">In another mode, it</td><td>provides</td><td>a</td><td></td><td>composition</td>

<td>pharmaceutical that</td><td>understands</td><td colspan="2">a composition</td><td>what</td><td>has the</td>

<td>structure (VI),</td><td colspan="2">or a stereoisomer,</td><td>Salt</td><td>or</td><td>prodrug</td>

<td>pharmaceutically</td><td>acceptable</td><td>of the same,</td><td>Y</td><td>a</td><td>vehicle,</td>

pharmaceutically acceptable diluent or excipient.

In another embodiment, a method of using a composition having structure (VI) in therapy is provided. In particular, the present description provides a method for treating cancer in a mammal, which comprises administering to an mammal in need thereof an effective amount of a composition having structure (VI) or a pharmaceutical composition comprising a composition having the structure (VI) and a pharmaceutically acceptable carrier, diluent or excipient.

In another embodiment, the present description provides a method for inhibiting tumor growth in a mammal that comprises administering to a mammal in need thereof an effective amount of a composition having structure (VI) or a pharmaceutical composition comprising a composition that It has structure (VI) and a pharmaceutically acceptable carrier, diluent or excipient.

In another embodiment, the present description provides a method for killing cancer cells in vitro using a composition having the structure (VI). In another embodiment, the present description provides a method for killing cancer cells in vivo in a mammal, which comprises administering to an mammal in need thereof an effective amount of a composition having structure (VI) or a pharmaceutical composition comprising a composition having structure (VI) and a pharmaceutically acceptable carrier, diluent or excipient.

In another embodiment, the present description provides a method for increasing the survival time of a mammal having cancer, which comprises administering to an mammal in need thereof an effective amount of a composition having structure (VI) or a pharmaceutical composition. which comprises a composition having structure (VI) and a pharmaceutically acceptable carrier, diluent or excipient.

These and other aspects of the disclosure will be apparent upon reference to the following detailed description.

BRIEF DESCRIPTION OF THE FIGURES

Figure 1 shows the (EC₅q) for each cell phone (HCC1954 and Jurkat).

Figure 2 shows a graph for Compound A in two Jurkat).

Figure 3 shows a graph for Compound B in two Jurkat).

Figure 4 shows a graph for Compound C in two Jurkat).

Figure 5 shows a graph cytotoxicity summary data

Compods Α-Ε for two lines of cytotoxicity data cell lines (HCC1954 and cytotoxicity data cell lines (HCC1954 and cytotoxicity data cell lines (HCC1954 and cytotoxicity data for Compound D on two cell lines (HCC1954 and

Jurkat)

Figure 6 shows a graph of cytotoxicity data for Compound E in two cell lines (HCC1954 and Jurkat).

Figure 7 shows a cell death curve in HCC1954 cells in vitro with the antibody-drug conjugates: T-LC-

<td>SPDP-A</td><td>(Trastuzumab,</td><td>LC-SPDP linker,</td><td>Compound</td><td>A) and</td><td>T-</td>

<td>SMCC-A</td><td>(Trastuzumab,</td><td>SMCC linker,</td><td>Compound</td><td>TO) .</td><td>The</td>

<td>values</td><td>EC50 is shown</td><td>ran in the figure.</td><td></td><td></td><td></td>

<td colspan="2">Figure 8 shows</td><td>a death curve</td><td>cell in</td><td colspan="2">cells</td>

<td>HCC1954</td><td>in vitro with</td><td>the conjugates ant</td><td colspan="2">icibody-drug:</td><td>T-</td>

<td>SPDP-B</td><td>(trastuzumab,</td><td>LC-SPDP linker,</td><td>Compound</td><td>B) and</td><td>T-</td>

<td>SMCC-A</td><td>(Trastuzumab,</td><td>SMCC linker,</td><td>Compound</td><td>B).</td><td>The</td>

EC50 values ​​are shown in the figure.

Figure 9 shows a cell death curve in HCC1954 cells in vitro with the antibody-drug conjugate: TSPDP-C (Trastuzumab, LC-SPDP linker, Compound C). The EC50 value is shown in the figure.

Figure 10 shows a cell death curve of HCC1954 cells in vitro with the antibody-drug conjugates: TMCvcPABC-85 (Trastuzumab, MCvc PABC linker, Compound 85), T-MCvcPABC-77 (Trastuzumab, MCvc PABC linker, Compound 77) and T-MCvcPABC-80 (Trastuzumab, MCvc PABC linker,

Compound 80). EC values₅₀ They are shown in the figure.

Figure 11 shows a cell death curve in BxPC-3 cells in vitro with the antibody-drug conjugate C-MCvcPABC-77, (Cetuximab, MCvc PABC linker, Compound 77), and a cell death curve in HPAF- cells. II in vitro with the antibody-drug conjugate C-MCvcPABC-77, (Cetuximab, MCvc PABC linker, Compound 77). EC values₅₀ They are shown in the figure.

Figure 12 shows a cell death curve of HCC1954 cells in vitro with the antibody-drug conjugates: TMCvcPABC-77 (Trastuzumab, MCvc PABC linker, Compound 77), T-MCvcPABC-85 (Trastuzumab, MCvc PABC linker, Compound 85) T-MCvcPABC-58 (Trastuzumab, MCvc PABC linker, Compound 58) and T-MCvcPABC-63 (Trastuzumab, MCvc PABC linker, Compound 63). EC50 values ​​are shown in the figure.

Figure 13 shows a cell death curve in NCI-N87 cells in vitro with the antibody-drug conjugates: TMCvcPABC-77, (Trastuzumab, MCvc PABC linker, Compound 77), T-MCvePABC-63, (Trastuzumab, MCvc PABC linker , Compound 63), T-MCvcPABC-85, (Trastuzumab, MCvc PABC linker, Compound 85), T-MCvcPABC-77, (Trastuzumab, linker

MCvc PABC, Compound 77), and T-MCvcPABC-80 (Trastuzumab,

<td>MCvc linker</td><td colspan="2">PABC, Compound</td><td> 80) .</td><td>The values</td><td>EC50</td><td>I know</td>

<td>show in the</td><td>figure.</td><td></td><td></td><td></td><td></td><td></td>

<td>Figure 14</td><td>show the</td><td colspan="3">live results</td><td>from</td><td>the</td>

<td>administration</td><td>of the Compound</td><td>F,</td><td>the</td><td>Compound 14,</td><td> 0</td><td>the</td>

Compound 23 in tumor volume in nude female nude mice with established tumors.

Figure 15 shows the live results of administration of the antibody-drug conjugate T-MCC-DM1 (Trastuzumab, MCC linker, DM1 maitansinoid) in varied doses as indicated, or T-MCvcPABC-77 at varied doses as indicated. indicates, in the volume of the tumor in female mice NOD / SCID Gamma with established tumors.

Figure 16 shows the live results of administration of the antibody-drug conjugate T-MCvcPABC-63 at 3 mg / kg, or T-MCvcPABC-77 at 3 mg / kg, in tumor volume in female NOD / SCID mice Gamma with established tumors.

Figure 17 shows a cell death curve in HCC1954 cells in vitro with the antibody-drug conjugates: TSPDP-140 (Trastuzumab, LC-SPDP linker, Compound 140) and TSMCC-140 (Trastuzumab, SMCC linker, Compound 140). Compound 140 is linked through the side chain of its N-terminal amino acid. EC50 values ​​are shown in the figure.

Figure 18 shows a cell death curve in HCC1954 cells in vitro with antibody-drug conjugates: T-SPDP142 (Trastuzumab, LC-SPDP linker, Compound 142) and T-SMCC142 (Trastuzumab, SMCC linker, Compound 142). Compound 142 is linked through the side chain of its N-terminal amino acid. EC values₅₀ They are shown in the figure.

Figure 19 shows a cell death curve in HCC1954 cells in vitro with antibody-drug conjugates: TMCvcPABC-58, (Trastuzumab, MCvc PABC linker, Compound 58), and T-MCvePABC-41, (Trastuzumab, MCvc PABC linker, Compound 41), and shows a cell death curve in NCI-N87 cells in vitro with the antibody-conjugated conjugates: T-MCvcPABC-58, (Trastuzumab, MCvc PABC linker, Compound 58), and T-MCvePABC-41, (Trastuzumab, linker MCvc PABC, Compound 41). Compound 41 is linked through the side chain of its N-terminal amino acid. Compound 58 is linked through the side chain of its amino acid Nterminal. EC values₅or are shown in the figure. EC values₅₀ They are shown in the figure.

DETAILED DESCRIPTION OF THE INVENTION

In the following description, certain specific details are set forth in order to provide a complete understanding of various modalities of this specification. However, one skilled in the art will understand that the present specification can be implemented without these details.

Unless the context requires otherwise, throughout the present specification and claims, the word includes and variations thereof, such as, includes and understands must be interpreted in an open, and inclusive sense, that is, including, but not limited to.

The reference, throughout this specification, to a modality (one embodiment) or a modality (an embodiment) means that a particular feature, structure or feature described in connection with the modality is included in at least one modality of the present specification . Therefore, the occurrences of the phrases in one modality (in one embodiment) or in one modality (in an embodiment) in various places throughout this specification are not necessarily all that refer to the same modality. In addition, particular features, structures, or characteristics may be combined in any suitable manner in one or more modalities.

Unless otherwise indicated, the following terms and phrases as used herein are intended to have the following meanings. When trade names (trademarks) are used herein, applicants intend to independently include the formulation of the brand name product, the generic drug, and the active pharmaceutical ingredient (s) ( s) of the brand product.

Amino refers to the substituent -NH₂.

Cyano refers to the substituent -CN.

Hydroxy or hydroxyl refers to the substituent -OH.

Imino refers to the substituent = NH.

Nitro refers to the substituent -NO₂.

Oxo refers to the substituent = 0.

Thiol refers to the substituent -SH.

Thioxo refers to the substituent = S.

Alkyl refers to a linear or branched hydrocarbon chain substituent consisting solely of carbon and hydrogen atoms, which is saturated or unsaturated (i.e., it contains one or more double and / or triple bonds), which has one to twelve atoms. carbon (Ci ~ Ci alkyl₂), preferably from one to eight carbon atoms (alkyl of

Ci-Cg) or one to six carbon atoms (Ci ~ C alkyl₆), and which is attached to the rest of the molecule by a single bond, for example, methyl, ethyl, n-propyl, 1-methylethyl (iso-propyl), n-butyl, n-pentyl, 1,1-dimethylethyl ( t-butyl), 3-methylhexyl, 2-methylhexyl, ethenyl, prop-l-enyl, but-l-enyl, pent-l-enyl, penta-1,4-dienyl, ethynyl, propynyl, butynyl, pentinyl, hexinyl, and Similar. Unless otherwise specifically indicated in the specification, an alkyl group may be optionally substituted.

"Alkylene" or "alkylene chain" refers to a linear or branched divalent hydrocarbon chain that binds the rest of the molecule to a substituent group, consisting solely of carbon and hydrogen, which is saturated or unsaturated (i.e. it contains one or more bonds doubles and / or triples), and having one to twelve carbon atoms, for example, methylene, ethylene, propylene, n-butylene, ethenylene, propenylene, n-butenylene, propynylene, n-butythylene, and the like. The alkylene chain is attached to the rest of the molecule through a single or double bond and to the substituent group through a single or double bond. The points of attachment of the alkylene chain with the rest of the molecule and to the substituent group may be through a carbon or any two carbons within the chain. Unless specifically indicated otherwise in the specification, an alkylene chain may be optionally substituted.

Alkoxy refers to a substituent of the formula

OR_to where R_to it is an alkyl substituent as defined above containing one to twelve carbon atoms. Unless otherwise specifically indicated in the specification, an alkoxy group may be optionally substituted.

Alkylamino refers to a substituent of the formula NHR_to or -NR_toR_to where every R_to it is, independently, an alkyl substituent as defined above containing from one to twelve carbon atoms. Unless otherwise specifically indicated in the specification, an alkylamino group may be optionally substituted.

Thioalkyl refers to a substituent of the formula -SR_towhere R_to it is an alkyl substituent as defined above containing from one to twelve carbon atoms. Unless otherwise specifically indicated in the specification, a thioalkyl group may be optionally substituted.

Aryl refers to a hydrocarbon substituent ring system comprising hydrogen, from 6 to 18 carbon atoms and at least one aromatic ring. For the purposes of this description, the aryl substituent may be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which may include fused or bridged ring systems. The aryl substituents include, but are not limited to, aryl substituents derived from aceantrylene, acenaphthylene, acefenantrylene, anthracene, blue, benzene, chrynene, fluorantene, fluorene, asindacene, s-indacene, indane, indene, naphthalene, phenanene, phenanthrene, pleiadene , pyrene and triphenylene. Unless otherwise specifically indicated in the specification, the term aryl or the prefix ar- (as in aralkyl) is intended to include aryl substituents that are optionally substituted.

Aralkyl refers to a substituent of the formula -R_b-R_cwhere R_b is an alkylene chain as defined above and R_c it is one or more aryl substituents as defined above, for example, benzyl, diphenylmethyl and the like. Unless otherwise specifically indicated in the specification, an aralkyl group may be optionally substituted.

Cycloalkyl or carbocyclic ring refers to a stable non-aromatic monocyclic or polycyclic hydrocarbon substituent consisting solely of carbon and hydrogen atoms, which may include fused or bridged ring systems, having three to fifteen carbon atoms, preferably having three to ten carbon atoms, and that is saturated or unsaturated and attached to the rest of the molecule by a single bond. Monocyclic substituents include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Polycyclic substituents include, for example, adamantyl, norbornyl, decalinyl, 7,7-dimethylbicyclo [2.2.1] heptanyl, and the like. Unless otherwise specifically indicated in the specification, a cycloalkyl group may be optionally substituted.

Cycloalkylalkyl refers to a substituent of the formula -R_bRd θη where R_d it is an alkylene chain as defined above and R_g It is a cycloalkyl substituent as defined above. Unless otherwise specifically indicated in the specification, a cycloalkylalkyl group may be optionally substituted.

Merging refers to any ring structure described herein that is fused to a ring structure in the compounds of the specification. When the fused ring is a heterocyclyl ring or a heteroaryl ring, any carbon atom in the existing ring structure, which becomes part of the fused heterocyclyl ring or the fused heteroaryl ring can be replaced with a nitrogen atom.

Halo or halogen refers to bromine, chlorine, fluorine or iodine. Haloalkyl refers to an alkyl substituent, as defined above, which is substituted by one or more halo substituents, as defined above, for example, trifluoromethyl, difluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,2- difluoroethyl, 3-bromo-2fluoropropyl, 1,2-dibromoethyl, and the like. Unless otherwise specifically indicated in the specification, a haloalkyl group may be optionally substituted.

Heterocyclic or heterocyclic ring refers to a non-aromatic substituent ring of 3 to 18 stable members consisting of two to twelve carbon atoms and one to six heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur. Unless otherwise specifically indicated in the specification, the heterocyclyl substituent may be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which may include fused or bridged ring systems; and the nitrogen, carbon or sulfur atoms in the heterocyclyl substituent may optionally be oxidized; the nitrogen atom may be optionally quaternized; and the heterocyclyl substituent may be partially or fully saturated. Examples of such heterocyclyl substituents include, but are not limited to, dioxolanyl, thienyl [1,3] dithianyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopyrazin-2-oxopyrazin-2-oxopyrazin-2-oxopyrazin-2-oxopyrazin-2-oxopyrazin-2-oxopyrazin-2-oxopyrazin-2-oxopyrazinyl-2-oxopyrazinyl-2-oxopyrazinyl-2-oxopyrazyl-2-oxopyrazyl-2-oxopyrazyl-2-oxopyrazinyl-2-oxopyrazinyl-2-oxopyrazinyl-2-Oxopyrazinyl-2-Oxopyrazinyl-2-Oxopyrazinyl-2-Oxopyrazinyl-2-Oxopyrazinyl-2-Oxide substituents. , piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl. Unless otherwise specifically indicated in the specification, a heterocyclyl group may be optionally substituted.

N-heterocyclyl refers to a heterocyclyl substituent as defined above that contains at least one nitrogen and where the point of attachment of the heterocyclyl substituent to the rest of the molecule is through a nitrogen atom in the heterocyclyl substituent. Unless otherwise specifically indicated in the specification, an N-heterocyclyl group may be optionally substituted.

Heterocyclylalkyl refers to a substituent of the formula -R_bR_and where R_b is an alkylene chain as defined above and R_and it is a heterocyclyl substituent as defined above, and if the heterocyclyl is a nitrogen-containing heterocyclyl, the heterocyclyl may be attached to the alkyl substituent on the nitrogen atom. Unless specifically indicated differently in the specification, a heterocyclylalkyl group may be optionally substituted.

"Heteroaryl" refers to a 5 to 14 substituent ring system comprising hydrogen atoms, one to thirteen carbon atoms, one to six heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur, and at least one aromatic ring . For the purposes of this description, the heteroaryl substituent may be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which may include fused or bridged ring systems; and the nitrogen, carbon or sulfur atoms in the heteroaryl substituent may optionally be oxidized; The nitrogen atom may be optionally quaternized. Examples include, but are not limited to, azepinyl, acridinyl, benzimidazolyl, benzothiazolyl, benzindolyl, benzodioxolyl, benzofuranyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, benzofb] [1,4] dioxepinyl,

1,4-benzodioxanil, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzothienyl (benzothiophenyl), benzotriazolyl, benzo [4,6] imidazoyl, dibinyl, dibinyl, dibinyl, dibinyl, dibinyl, dibinyl, dibinyl, dibinyl, dibinyl, dibinyl, dibyl, nyl, benzyl [dothyl], dothizyl] , dibenzothiophenyl, furanyl, furanonyl, isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl, isoquinolyl, indolizinyl, isoxazolyl, naphthyridinyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl, 1-oxidopyridinyl, 1-oxidopirimidinilo, 1-oxidopirazinilo, 1-oxidopiridazinilo, 1-phenyl-lfl-pyrrolyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, quinazolinyl, quinoxalinyl, quinolinyl, quinuclidinyl, isoquinolinyl, tetrahydroquinolinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, and thiophenyl (i.e. thienyl). Unless otherwise specifically indicated in the specification, a heteroaryl group may be optionally substituted.

N-heteroaryl refers to a heteroaryl substituent as defined above that contains at least one nitrogen and where the point of attachment of the heteroaryl substituent to the rest of the molecule is through a nitrogen atom in the heteroaryl substituent. Unless otherwise specifically indicated in the specification, an N-heteroaryl group may be optionally substituted. Heteroarylalkyl refers to a substituent of the formula -RbRf where R_b is an alkylene chain as defined above and R_F it is a heteroaryl substituent as defined above. Unless otherwise specifically indicated in the specification, a heteroarylalkyl group may be optionally substituted.

The substituted term used herein means any of the above groups {ie, alkyl, alkylene, alkoxy, alkylamino, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, haloalkyl, heterocyclyl, N-heterocyclyl, heterocyclylalkyl, heteroaryl, Nheteroaryl and / or heteroarylalkyl) wherein at least one hydrogen atom is replaced by a bond to an atom other than hydrogen, such as, but not limited to: a halogen atom such as F, Cl, Br, and I; an oxygen atom in groups such as hydroxyl groups, alkoxy groups, and ester groups; a sulfur atom in groups such as thiol groups, thioalkyl groups, sulfone groups, sulfonyl groups and sulfoxide groups; a nitrogen atom in groups such as azides, amines, amides, alkylamines, dialkylamines, arylamines, alkylamines, diarylamine, N-oxides, imides, and enamines, · a silicon atom in groups such as trialkylsilyl groups, dialkylarylsilyl groups, alkyldyrylsilyl groups , and triarylsilyl groups; and other heteroatoms in various other groups. Substituted also means any of the above groups in which one or more hydrogen atoms are replaced by a higher order bond (eg, double or triple bond) to a heteroatom such as oxygen in oxo, carbonyl, carboxyl, and ester groups ; and nitrogen in groups such as imines, oximes, hydrazones, and nitriles. For example, substituted includes any of the above groups in which one or more hydrogen atoms are replaced with -NR_gR_h, -NR_gC (= O) R_h, -NR_gC (= 0) NR_gR_h, -NR_gC (= 0) OR_h, -NR_gC (= N Rg) NR_gR_h, -NRgSO₂R_h, -OC (= 0) NRgR_h, -0R_g, -MR_g, -S0R_g, -SO₂R_g, -OS 0₂R_g, -SO₂ORg, = NSO₂Rg, and -S0₂NR_gR_h. Substituted also means any of the above groups in which one or more hydrogen atoms are replaced with -C (= O) R_g, -C (= O) OR_g, -C (= O) NR_gR_h, -CH₂SW₂R_g, -CH₂SW₂NR_gR_h. In the foregoing, R_g and R_h hydrogen, alkyl, alkoxy, alkylamino, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, haloalkyl, heterocyclyl, N heterocyclyl, heterocyclylalkyl, heteroaryl, Nheteroaryl and / or heteroarylalkyl are the same or different.

Substituted also means any of the above groups in which one or more hydrogen atoms are replaced by a link to an amino, cyano, hydroxyl, imino, nitro, oxo, thioxo, halo, alkyl, alkoxy, alkylamino, thioalkyl, aryl group , aralkyl, cycloalkyl, cycloalkylalkyl, haloalkyl, heterocyclyl, N-heterocyclic, heterocyclylalkyl, heteroaryl, N-heteroaryl and / or heteroarylalkyl. In addition, each of the above substituents may also be optionally substituted with one or more of the above substituents.

The term "protecting group", as used herein, refers to a labile chemical residue that is known in the art that protects reactive groups including, without limitation, hydroxyl and amino groups, against unwanted reactions during synthetic procedures. The hydroxyl and amino groups that are protected with a protecting group are referred to herein as protected hydroxyl groups and protected amino groups, respectively. Protective groups are normally used selectively and / or orthogonally to protect sites during reactions at other reactive sites and can then be removed to leave the group unprotected as is or available for other reactions. Protective groups as known in the art are generally described in Greene and Wuts, Protective Groups in Organic Synthesis, 3rd edition, John Wiley &amp; Sons, New York (1999). The groups can be selectively incorporated into the compounds of the present description as precursors. For example, an amino group can be placed in a compound of the description as an azido group that can be chemically converted to the amino group at a desired point in the synthesis. In general, the groups are protected or present as a precursor that will be inert to reactions that modify other areas of the parental molecule for conversion into their final groups at an appropriate time. Representative additional protecting groups or precursors are discussed in Agrawal, et al., Protocols for Oligonucleotide Conjugates, Eds, Humana Press; New Jersey, 1994; Vol. 26 pp 1-72. Examples of hydroxyl protecting groups include, but are not limited to, t-butyl, t-butoxymethyl, methoxymethyl, tetrahydropyranyl, 1ethoxyethyl, 1- (2-chloroethoxy) ethyl, 2-trimethylsilylethyl, pclorophenyl, 2,4-dinitrophenyl, benzyl, 2 , 6-dichlorobenzyl, diphenylmethyl, p-nitrobenzyl, triphenylmethyl, trimethylsilyl, triethylsilyl, t-butyldimethylsilyl, t-butyldiphenylsilyl (TBDPS), triphenylsilyl, benzoylformate, acetate, chloroacetate, trichloroacetate, benzoyl acetate p-phenylbenzoate, 9fluorenylmethyl carbonate, mesylate and tosylate. Examples of amino protecting groups include, but are not limited to, carbamate protecting groups, such as 2-trimethylsilylethoxycarbonyl (Teoc), 1-methyl-l- (4-biphenylyl) ethoxycarbonyl (Bpoc), t-butoxycarbonyl (BOC), allyloxycarbonyl (Alloc), 9-fluorenylmethyloxycarbonyl (Fmoc), and benzyl-oxycarbonyl (Cbz); amide protecting groups, such as formyl, acetyl, trihaloacetyl, benzoyl, and nitrophenylacetyl; sulfonamide protecting groups, such as 2-nitrobenzenesulfonyl; and imine and cyclic imide protecting groups, such as phthalimide and dithiasuccinoyl.

Prodrug is intended to indicate a compound that can be converted under physiological conditions or by sololysis into a biologically active compound of the present disclosure. Therefore, the term "prodrug" refers to a metabolic precursor of a compound of the present disclosure that is pharmaceutically acceptable. A prodrug may be inactive when administered to a subject in need thereof, but is converted in vivo to an active compound of the specification. In one embodiment, a prodrug is rapidly transformed in vivo to give the parent compound of the description, for example, by blood hydrolysis. In one embodiment, a prodrug can be stable in the plasma or blood. In one embodiment, a prodrug can be the objective (target) form of a compound of the invention. The prodrug compound often offers advantages of solubility, tissue compatibility or delayed release in a mammalian organism (see, Bundgard, H. , Design of Prodrugs (1985), pp. 7-9, 21-24 (Elsevier, Amsterdam)). A discussion of prodrugs is provided in Higuchi, T., et al., ACS Symposium Series, Vol. 14, and in Bioreversible Carriers in Drug Design, Ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987.

The term prodrug is understood to include any covalently bound vehicle, which releases the active compound of the present description in vivo when said prodrug is administered to a mammalian subject. Conjugates, including ADCs, as described herein, are such prodrugs of compositions having structure (I), (la) or (Ib). Prodrugs of a compound of the description can be prepared by modifying functional groups present in the compound of the description in such a way that the modifications are cleaved, either in routine manipulation or in vivo, in the parental compound of the present description. Prodrugs include compounds of the description wherein, the hydroxy, amino or mercapto group is attached to any group that, when the prodrug of the compound of the description is administered to a mammalian subject, is cleaved to form a free hydroxy group, a group free amino or a free mercapto group, respectively. Examples of prodrugs include, but are not limited to, acetate, formate and benzoate derivatives of alcohol or amide derivatives of amine functional groups in the compounds of the present disclosure and the like.

The present description is also intended to cover all pharmaceutically acceptable compounds of structure (I), (la) or (Ib) that have been isotopically labeled by having one or more atoms replaced by an atom that has an atomic mass or mass number different. Examples of isotopes that can be incorporated into the disclosed compounds include hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, chlorine and iodine isotopes, such as²H ³H ^{1: L}C, ¹³C, ¹⁴C, ¹³N, ¹⁵N, ¹⁵0, ¹⁷0, ¹⁸0, ³¹P, ³²P, ³⁵S, ¹⁸F, ³⁶C1, ¹²³I, and ¹²⁵I, respectively. These radiolabeled compounds could be useful to help determine or measure the efficacy of the compounds, by characterizing, for example, the site or mode of action, or the binding affinity to the pharmacologically important site of action. Certain isotopically labeled compounds of structure (I), (la) or (Ib), for example, those that incorporate a radioactive isotope, are useful in drug studies and / or tissue distribution of the substrate. Tritium radioactive isotopes, that is, ³H, and carbon-14, that is, ¹⁴C, are particularly useful for this purpose in view of their ease of incorporation and rapid detection means.

Substitution with heavier isotopes such as deuterium, that is, ²H, may provide certain therapeutic advantages resulting from increased metabolic stability, for example, the increase in half-life in vivo or reduced dosage requirements, and therefore may be preferred in some circumstances.

Substitution with positron emitting isotopes, such as ⁿC, ¹⁸F, ¹⁵0 Y ¹³N, they may be useful in Positron Emission Topography (PET) studies to examine the receptor substrate occupation. Isotopically labeled compounds of structure (I), (la) or (Ib) can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described in the Preparations and Examples set forth below using a suitable reagent isotopically labeled instead of the unlabeled reagent previously used.

The present description is also intended to cover in vivo metabolic products of the described compounds. Such products may be the result of, for example, the oxidation, reduction, hydrolysis, amidation, esterification, and the like of the compound administered, mainly due to enzymatic processes. Accordingly, the present description includes compounds produced by a process comprising administering a compound of this description to a mammal for a period of time sufficient to produce a metabolic product thereof. Such products are typically identified by administration of a radiolabeled compound of the description in a dose detected to an animal, such as the rat, mouse, guinea pig, monkey, or human, leaving sufficient time for metabolism to occur, and isolating its products. conversion of urine, blood or other biological samples.

Stable compound and stable structure are intended to indicate a compound that is sufficiently robust to survive isolation to a useful degree of purity of a reaction mixture, and formulation in an effective therapeutic agent.

The term "antibody" herein is used in the broadest sense and specifically covers intact monoclonal antibodies, polyclonal antibodies, multispecific antibodies (eg, bispecific antibodies) formed from at least two intact antibodies, and antibody fragments, provided that exhibit the desired biological activity. The term "antibody" refers to a full-length immunoglobulin molecule or a functionally active portion of a full-length immunoglobulin molecule, that is, a molecule that contains an antigen binding site that immunospecifically binds to a target antigen. of interest or part thereof. The immunoglobulin described herein can be of any type (eg, IgG, IgE, IgM, IgD, and IgA), class (eg, IgGl, IgG2, IgG3, IgG4, igAl and IgA2) or subclass of immunoglobulin molecule. Immunoglobulins can be derived from any species. In one aspect, the immunoglobulin is of human, murine, or rabbit origin. In another aspect, the antibodies are polyclonal, monoclonal, multi-specific (e.g. bispecific), human, humanized or chimeric antibodies, linear antibodies, single chain antibodies, diabodies, maxybodies, minibodies, Fv, Fab fragments, F fragments (ab '), F (ab') fragments₂, fragments produced by a Fab expression library, anti-idiotypic (anti-Id) antibodies, CDRs, and epitope binding fragments of any of the above that bind immunospecifically to a target antigen.

The term monoclonal antibody as used herein,

<td>it means</td><td>to an antibody</td><td>obtained from</td><td>a</td><td>population</td><td>from</td>

<td>antibodies</td><td>substantially</td><td>homogeneous,</td><td>it is</td><td>tell,</td><td>the</td>

<td>antibodies</td><td>individual that</td><td>understand</td><td>the</td><td>population</td><td>They are</td>

identical except for possible naturally occurring mutations that may be present in smaller amounts. Monoclonal antibodies include chimeric antibodies in which a portion of the heavy and / or light chain is identical to or homologous to the corresponding sequences in antibodies derived from a particular species or belonging to a particular class or subclass of antibody, while the rest of the chain (s) is identical with or homologous to the corresponding sequences in antibodies derived from other species or belonging to another class or subclass of antibody, as well as fragments of such antibodies (see, for example, US Pat. No. 4,816,567; and Morrison et al., 1984, Proc. Nati.

Acad. Sci. USA 81: 6851-6855). Monoclonal antibodies also include humanized antibodies that can contain a completely human constant reaction and a CDRs from a non-human source.

An intact antibody is one that comprises a variable region of antigen binding, as well as a light chain constant domain (CL) and heavy chain constant domains,

<td>Chi &lt;c_Htwo Y</td><td>C_h3</td><td colspan="3">Constant domains</td><td>they can</td><td>be</td><td>domains</td>

<td>constants</td><td>from</td><td>sequence</td><td>native</td><td>(by</td><td colspan="2">example,</td><td>domains</td>

<td>constants</td><td>from</td><td>sequence</td><td>native</td><td colspan="2">human) or</td><td colspan="2">variants of</td>

amino acid sequence thereof.

The antibody fragments comprise a portion of an intact antibody, which preferably comprises the antigen-binding or variable reqion thereof. Examples of antibody fragments include Fab, Fab ', F (ab') 2f and Fv fragments; diabody linear antibodies; single chain antibody molecules; maxybodies; minibodies; and multispecific antibodies formed from antibody fragment (s).

An isolated antibody is one that has been identified and separated and / or recovered from a component of its natural environment. Contaminating components of their natural environment are materials that would interfere with diagnostic or therapeutic uses for the antibody, and may include enzymes, hormones, and other protein or non-protein solutes. In some embodiments, the antibody will be purified (1) to more than 95% by weight of antibody as determined by Lowry's method, and more preferably more than 99% by weight, (2) to a degree sufficient to obtain at least 15 amino acid residues of the N-terminal or internal sequence through the use of a rotating cup sequencer, or (3) until homogeneity by SDS-PAGE under reducing or non-reducing conditions using Coomassie blue or preferably, silver staining The isolated antibody includes the antibody in situ within recombinant cells since at least one component of the natural environment of the antibody will not be present. Usually, however, the isolated antibody will be prepared by at least one purification step.

An antibody that binds to an antigen of interest is one capable of binding to that antigen with sufficient affinity such that the antibody is useful for targeting a cell that expresses the antigen.

A native sequence polypeptide is one that has the same amino acid sequence as a nature derived polypeptide. Such native sequence polypeptides can be isolated from nature or can be produced by recombinant or synthetic means. Therefore, a native sequence polypeptide can have the amino acid sequence of naturally occurring human polypeptides, murine polypeptides or polypeptides of any other mammalian species.

The term "intracellular metabolite" refers to a compound resulting from a metabolic process or reaction within a cell in a composition of the invention (for example, a drug-antibody conjugate (ADC)). The metabolic process or reaction may be an enzymatic process such as the proteolytic cleavage of a linker peptide of the composition in question, or the hydrolysis of a functional group such as a hydrazone, ester, or amide within the composition in question. In the context of conjugates, including ADCs, intracellular metabolites include, but are not limited to, antibodies and free drugs that have been separated intracellularly, that is, after admission, diffusion, absorption or transport into a cell (e.g., by enzymatic cleavage of an ADC by an intracellular enzyme).

In the context of conjugates, including ADCs, the terms intracellularly cleaved and intracellular cleavage refer to metabolic processes or reactions within a cell in a composition of the invention in which the covalent bond, for example, the linker (L), between the drug residue (D) and the target residue (T) (for example, an antibody) is broken, resulting in the free drug dissociated from (T) within the cell. In one embodiment, the cleaved residues of the present compositions are therefore intracellular metabolites (eg, T, TL fragment, DL fragment, D). Accordingly, in one embodiment, the invention provides compositions that are cleavage products of a composition having the structure (VI), that the cleavage products include compositions comprising the structure (I), (la) or (Ib), or stereoisomers thereof. Similarly, the linker (L), between the microtubule disrupting peptide toxin (PT) and the target residue (T) (eg, an antibody) can be broken intracellularly, resulting in the PT dissociation of (T) within of the cell. The cleaved residues of the present compositions are therefore intracellular metabolites (eg, T, TL fragment, PT-L fragment, PT). Accordingly, in one embodiment, the invention provides compositions that are cleavage products of a composition having structure (VII), whose cleavage products include compositions of structure (I), (la) or (Ib), or stereoisomers of the same.

The term "extracellular cleavage" refers to a metabolic process or reaction outside a cell in a composition of the invention in which the covalent bond, for example, the linker (L), between the drug residue (D) and the target residue (T) (for example, an antibody) is broken, resulting in the free drug dissociated from (T) outside the cell. In one embodiment, the cleaved residues of the present compositions are therefore initially extracellular metabolites (eg, T, TL fragment, DL fragment, D), which can be moved intracellularly by diffusion and cell permeability or transport. Accordingly, in one embodiment, the invention provides compositions that are cleavage products of a composition having structure (VI), whose cleavage products include compositions comprising structure (I), (la) or (Ib), or stereoisomers thereof. Similarly, the linker (L), between the microtubule disrupting peptide toxin (PT) and the target residue (T) (for example, an antibody) can be broken extracellularly, resulting in the dissociated PT from (T) outside of the cell. The cleavage residues of the present compositions are therefore extracellular metabolites initially (eg, T, TL fragment, PT-L fragment, PT). Accordingly, in one embodiment, the invention provides compositions that are cleavage products of a composition having structure (VII), whose cleavage products include compositions comprising structure (I), (la) or (Ib), or stereoisomers thereof.

Mammal includes both humans and domestic animals such as laboratory animals and domestic animals (for example, cats, dogs, pigs, cows, sheep, goats, horses, rabbits), and non-domestic animals, such as wildlife and the like .

Optionally or optionally means that subsequent events of circumstances described may or may not occur, and that the description includes cases in which said event or circumstance occurs and cases in which it does not occur. For example, optionally substituted aryl means that the aryl substituent may or may not be substituted and that the description includes both the substituted aryl substituents and aryl substituents which have no substitution.

Pharmaceutically acceptable diluent or excipient carrier includes without limitation any adjuvant, vehicle, excipient, slip agent, sweetening agent, diluent, preservative, pigment / dye, flavor enhancer, surface active agent, wetting agent, dispersing agent, suspending agent, stabilizer, isotonic agent, solvent or emulsioficante that has been approved by the Food and Drug Administration of the United States (or another regulatory agency similar to another jurisdiction) as acceptable for use in humans or pets. Pharmaceutically acceptable salt includes both acid addition salts and basic addition salts.

Pharmaceutically acceptable acid addition salt refers to those salts that retain the biological efficacy and properties of free bases, which are not biologically or otherwise undesirable, and which are formed with inorganic acids such as, but not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like, and organic acids such as, but not limited to, acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, canphoric acid, camfor-10-sulfonic acid, capric acid, caproic acid, caprylic acid, carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecyl sulfuric acid , ethan-1, 2 disulfonic acid ethanesulfonic acid

257 hydroxyethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid, gluconic acid, glucuronic acid, glutamic acid, glutaric acid, 2-oxo-glutaric acid, glycerophosphoric acid, glycolic acid, hipuric acid, isobutyric acid, lactic acid , lactobionic acid, lauric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, mucic acid, naphthalen-1,5-disulfonic acid, naphthalen-2-sulfonic acid, l-hydroxy-2-naphthic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, pyroglutamic acid, pyruvic acid, salicylic acid, 4-aminoalicylic acid, sebacic acid, stearic acid, succinic acid , tartaric acid, thiocyanic acid, ptoluenesulfonic acid, trifluoroacetic acid, undecylenic acid, and the like.

Pharmaceutically acceptable basic addition salt refers to those salts that retain the biological effectiveness and properties of free acids, which are not biologically or otherwise undesirable. These salts are prepared from the addition of an inorganic base or an organic base to the free acid. Salts derived from inorganic bases include, but are not limited to, the sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum and the like salts. Preferred inorganic salts are the ammonium, sodium, potassium, calcium, and magnesium salts. Salts derived from organic bases include, but are not limited to, salts of primary, secondary and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine , triethylamine, tripropylamine, diethanolamine, ethanolamine, deanol, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, benetamine, benzathine, ethylenediamine, glucosamine, methylglucamine, theobromine, triethanolamine, tromethamine, purines, piperazine, piperidine, N-ethylpiperidine, polyamine resins and the like. Particularly, the preferred organic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline and caffeine.

Frequent crystallizations produce a solvate of the compound of the description. As used herein, the term "solvate" refers to an aggregate comprising one or more molecules of a compound of the present specification with one or more solvent molecules. The solvent can be water, in which case the solvate can be a hydrate. Alternatively, the solvent may be an organic solvent. Therefore, the compounds of the present description may exist as a hydrate, including a monohydrate, dihydrate, hemihydrate, sesquihydrate, trihydrate, tetrahydrate and the like, as well as the corresponding solvated forms. The compound of the present description may be a true solvate, while in other cases, the compound of the present description may only retain adventitious water or be a mixture of water plus some adventitious solvent.

A pharmaceutical composition refers to a formulation of a compound of the present disclosure and a medium generally accepted in the art for the delivery of a biologically active compound to mammals, for example, humans. Such means includes all pharmaceutically acceptable carriers, diluents or excipients thereof.

Non-limiting examples of disorders to be treated herein include benign and malignant tumors; leukemia and malignant lymphoid tumors, particularly of breast, ovarian, stomach, endometrial, salivary gland, lung, kidney, colon, thyroid, pancreas, prostate or bladder cancer; neuronal, glial, astrocital, hypothalamic and other glandular, macrophage, epithelial, stromal and blastocoelic disorders, autoimmune disease, inflammatory disease, fibrosis, and infectious diseases. Given the characteristics, and in particular the potency of the compositions in question, it will be apparent to the expert of reasonable skill, that the compounds of the invention may be indicated for use in treating any disease in which it is desirable to exercise a cytostatic or cytotoxic effect on a target cell.

In one embodiment, the compositions of the invention are used to treat autoimmune diseases. Immunospecific antibodies to an antigen of a cell that is responsible for producing autoimmune antibodies can be obtained from any organization (for example, a University scientist or a company like Genentech) or produced by any method known to a person skilled in the art such such as chemical synthesis or recombinant expression techniques. In another embodiment, useful ligand antibodies that are immunospecific for the treatment of autoimmune diseases include, but are not limited to, anti-nuclear antibody; Anti ds DNA; anti ss DNA, IgM antibody, anti cardiolipin IgG; IgM antibody, anti phospholipid IgG; anti SM antibody; anti mitochondrial antibody; thyroid antibody; microsomal antibody; thyroglobuline antibody; anti SCL-70; AntiJo; Anti-UlRNP; Anti-La / SSB; anti SSA; anti SSB; anti perital cell antibody; anti histones; anti RNP; C-ANCA; PANCA; anti centromere; anti fibrillary and anti MBG antibody. In certain preferred embodiments, antibodies useful in the present methods can be bound to either a receptor or a receptor complex expressed in an activated lymphocyte.

The receptor or receptor complex may comprise a member of the immunoglobulin gene superfamily, a member of the TNF receptor superfamily, an integrin, a cytokine receptor, a chemokine receptor, a protein with higher histocompatibility, a lectin, or a complement control protein. Non-limiting examples of suitable members of the immunoglobulin superfamily are CD2, CD3, CD4, CD8, CD19, CD22, CD28, CD79, CD90, CD152 / CTLA-4, PD-1, and ICOS.

Non-limiting examples of suitable members of the TNF receptor superfamily are CD27, CD40, CD95 / Fas,

CD134 / OX40, CD137 / 4-1BB, TNF-R1, TNFR-2, RANK, TACI, BCMA, osteoprotegerin, Apo2 / TRAIL-R1, TRAIL-R2, TRAIL-R3, TRAIL-R4 and APO-3. Non-limiting examples of suitable integrins are CD1 la, CDllb, CDllc, CD18, CD29, CD41, CD49a, CD49b, CD49c, CD49d, CD49e, CD49f, CD103, and CD104. Non-limiting examples of suitable lectins are lectin type-C, type-S, and type-I.

In one embodiment, the ligand is an antibody that binds to an activated lymphocyte that is associated with an autoimmune disease.

Immune diseases that are characterized by inappropriate activation of immune cells and that can be treated or prevented by the methods described herein can be classified, for example, by the type (s) of hypersensitivity reaction (s) that underlie the disorder. These reactions are typically classified into four types: anaphylactic reactions, cytotoxic (cytolytic) reactions, immune complex reactions, or cell-mediated immunity reactions (MIC) (also referred to as delayed-type hypersensitivity reactions (DTH)). (See, for example, Fundamental Immunology (William E. Paul ed., Raven Press, NY, third ed. 1993)).

Specific examples of such immunological diseases include the following: rheumatoid arthritis, autoimmune demyelinating diseases (e.g., multiple sclerosis, allergic encephalomyelitis), endocrine ophthalmopathy, uveorretinitis, systemic lupus erythematosus, myasthenia gravis, Grave's disease, glomerulonephritis, autoimmune hepatological disorder, inflammatory bowel disease (example of Crohn's bowel disease (example of Crohn's inflammatory bowel disease) ), anaphylaxis, allergic reaction, Sjogren's syndrome, type I diabetes mellitus, primary biliary cirrhosis, Wegener's granulomatosis, fibromyalgia, polymyositis, dermatomyositis, multiple endocrine insufficiency, Schmidt syndrome, autoimmune uveitis, Addison's disease, adrenalitis, thyroiditis, Hashimoto's thyroiditis, autoimmune thyroid disease, pernicious anemia, gastric atrophy, chronic hepatitis, lupoid hepatitis, atherosclerosis, lupus erythema subacute, hypoparathyroidism, Dressler's syndrome, autoimmune thrombocytopenia, idiopathic thrombocytopenic purpura, hemolytic anemia, pemphigus vulgaris, pemphigus, dermatitis herpetiformis, alopecia arcata, pemphigoid, scleroderma, progressive systemic sclerosis, CREST syndrome (calcinosis, Raynaud's phenomenon, esophageal dysmotility, sclerodactyl and telangiectasia), male and female autoimmune infertility, ankylosing spondylitis, ankylosing disease, connective tissue disease , nedosa polyarteritis, systemic necrotizing vasculitis, atopic dermatitis, atopic rhinitis, Goodpasture syndrome, Chagas disease, sarcoidosis, rheumatic fever, asthma, recurrent abortion, antiphospholipid syndrome, farmer's lung, erythema multiforme, post cardiotomy syndrome, Cushing's syndrome, autoimmune chronic active hepatitis, paramoral breeder's lung, toxic epidermal necrolysis, Alport syndrome, alveolitis, alveolitis allergic, fibrosing alveolitis, interstitial lung disease, erythema nodosum, pyoderma gangrenosum, transfusion reaction, Takayasu arteritis, polymyalgia rheumatica, temporal arteritis, schistosomiasis, giant cell arteritis, ascariasis, aspergillosis, Sampler syndrome, eczema, lymphomatoid granulomatosis, Behcet's disease, Caplan syndrome, Kawasaki disease, dengue, encephalomyelitis, endocarditis, endomyocardial fibrosis, endophthalmitis, erythema, erythema, erythema , psoriasis, fetal erythroblastosis, eosinophilic fasciitis, Shulman syndrome, Felty's syndrome, filariasis, cyclitis, chronic cyclitis, heterochronous cyclitis, Fuch cyclitis, IgA nephropathy, Henoch-Schonlein purpura, graft versus host disease, transplant rejection, cardiomyopathy, EatonLambert syndrome, recurrent polychondritis, cryoglobulinemia, Waldenstrom macroglobulinemia, Evans syndrome, and autoimmune gonadal insufficiency. In accordance with the foregoing, the methods described herein encompass the treatment of B lymphocyte disorders (eg, systemic lupus erythematosus, Goodpasture's syndrome, rheumatoid arthritis, and type I diabetes), Thl lymphocytes (e.g. rheumatoid arthritis, multiple sclerosis, psoriasis, Sjorgren's syndrome, Hashimoto's thyroiditis, Grave's disease, primary biliary cirrhosis, Wegener's granulomatosis, tuberculosis, or acute graft versus host disease), or Th2 lymphocytes (for example, atopic dermatitis, systemic lupus erythematosus, atopic asthma, rhinoconjunctivitis, allergic rhinitis, Ornen syndrome, systemic sclerosis, or chronic graft versus host disease). In general, disorders related to dendritic cells involve disorders of Thl lymphocytes or Th2 lymphocytes.

In certain embodiments, the immune disorder is mediated by T cells, which may include activated T cells. ADCs or ADC derivatives can be administered to deplete such activated T cells.

In one embodiment, the compositions of the invention can be used to treat fibrosis. Fibrosis can occur in many tissues within the body, typically as a result of inflammation or damage, examples include but are not limited to: Lungs, pulmonary fibrosis, idiopathic pulmonary fibrosis, cystic fibrosis; Liver, liver cirrhosis; Heart, endomyocardial fibrosis, old myocardial infarction, atrial fibrosis; Others, mediastinal fibrosis (soft tissue of the mediastinum), myelofibrosis (bone marrow), retroperitoneal fibrosis (soft tissue of the retroperitoneum), progressive massive fibrosis (lungs); a complication of pneumoconiosis of coal workers, nephrogenic systemic fibrosis (skin), Crohn's disease (intestine), keloids (skin), systemic sclerosis / ecleroderma (skin, lungs), arthrofibrosis (knee, shoulder, other joints), Peyronie's disease (penis), Dupuytren's contracture (hands, fingers) and some forms of adhesive capsulitis (shoulder).

With respect to infectious diseases, the compositions of the invention can be used directly in certain infectious or pathogenic agents, or they can be used to exert a cytostatic or cytotoxic effect on a host cell that houses, or otherwise provides, the infectious agent or pathogen.

Effective amount or therapeutically effective amount refers to that amount of a compound of the description which, when administered to a mammal, preferably a human being, is sufficient to exercise the treatment, as defined below, of the particular indication (by example, cancer or tumor cells in the mammal, preferably a human). The amount of a compound of the description that constitutes a therapeutically effective amount will vary depending on the compound, the condition and its severity, the form of administration, and the age of the mammal to be treated, but can be routinely determined by a person with ordinary experience in the technique taking into account your own knowledge and this description.

Treating or treating as used herein covers the treatment of the disease or condition of interest in a mammal, preferably a human being, who has the disease or condition of interest, and includes:

<td>(i) prevent</td><td colspan="2">the disease occurs or</td><td>condition</td><td>in a</td>

<td>mammal in</td><td>particular when</td><td>saying</td><td>mammal</td><td>is</td>

<td>predisposed</td><td>to the condition but</td><td>what</td><td>not yet</td><td>It has been</td>

<td>diagnosed</td><td>have it;</td><td></td><td></td><td></td>

(ii) inhibit the disease or condition, that is, stop its development;

(iii) relieve the disease or condition, that is, cause regression of the disease or condition; or (iv) relieve symptoms resulting from the disease or condition, that is, relieve pain without addressing the underlying disease or condition.

<td colspan="2">An amount</td><td colspan="2">therapeutically effective</td><td>of the compound</td><td>in</td>

<td>relationship</td><td>with</td><td>the treatment of</td><td>Cancer</td><td>can reduce</td><td>the</td>

<td>number</td><td>from</td><td>cancer cells;</td><td>reduce</td><td>the size</td><td>of the</td>

tumor; inhibit (ie, slow to some extent and preferably stop) the infiltration of cancer cells in peripheral organs; inhibit (ie, slow to some extent and preferably stop) tumor metastasis; inhibit, to some extent, tumor growth; increase survival time; and / or relieve to some extent one or more of the symptoms associated with cancer. To the extent that the drug can prevent growth and / or kill existing cancer cells, it can be cytostatic and / or cytotoxic. The compounds of the present invention are preferably cytotoxic. For cancer therapy, efficacy can, for example, be measured by assessing the time to disease progression (TTP) and / or determining the response rate (RR).

An effective amount with respect to a particular result to be achieved is an amount sufficient to achieve the desired result. For example, an effective amount of the drug when referred to in relation to the death of cancer cells, refers to an amount of drug sufficient to produce the lethal effect.

Solid tumors contemplated for treatment using the compounds described in the present description include, but are not limited to: sarcoma, fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovoma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon cancer, colorectal cancer, kidney cancer, colorectal cancer bone cancer, breast cancer, ovarian cancer, prostate cancer, esophageal cancer, stomach cancer (e.g. gastrointestinal cancer), oral cancer, nasal cancer, throat cancer, squamous cell carcinoma (e.g., lung), basal cell carcinoma, adenocarcinoma (e.g., lung), sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinomas, cystadenocarcinoma, medullary carcinoma , bronchogenic carcinoma, renal cell carcinoma, biliary duct carcinoma hepatoma, choriocarcinoma, seminoma, embryonic carcinoma, Wilms tumor, cervical cancer, uterine cancer, testicular cancer, small cell lung carcinoma, bladder carcinoma, lung cancer, non-small cell lung cancer, epithelial carcinoma, glioma, glioblastoma multiform astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma meningioma, skin cancer, melanoma, neuroblastoma and retinoblastoma. Blood-borne cancers contemplated for treatment using the compounds described herein include, but are not limited to: ALL acute lymphoblastic leukemia, acute B-cell lymphoblastic leukemia, acute T-cell lymphoblastic leukemia, AML acute myeloblastic leukemia, APL acute promyelocytic leukemia, acute monoblastic leukemia, acute erythroleukemic leukemia, acute megakaryoblastic leukemia, acute myelomonocytic lymphocytic leukemia acute undifferentiated leukemia, CML chronic myelocytic leukemia, CLL chronic lymphocytic leukemia, hair cell leukemia, and multiple myeloma. Acute and chronic leukemias contemplated for treatment using the compounds described herein include, but are not limited to: lymphoblastic, myelogenous, lymphocytic and myelocytic leukemia. Lymphomas contemplated for treatment using the compounds described herein include, but are not limited to: Hodgkin's disease, non-Hodgkin's lymphoma, multiple myeloma, Waldenstrom's macroglobulinemia, heavy chain disease, and polycythemia vera. Other types of cancer contemplated for treatment using the compounds described herein include, but are not limited to: peritoneal cancer, hepatocellular cancer, hepatoma, salivary cancer, vulvar cancer, thyroid cancer, penile cancer, anal cancer, head cancer. and neck, renal cell carcinoma, acute anaplastic large cell carcinoma, and cutaneous anaplastic large cell carcinoma.

Cancers, including, but not limited to, a tumor, metastasis, or other disease or disorder characterized by uncontrolled or unwanted cell growth, can be treated or prevented by administering the compounds described herein.

In other embodiments, methods for treating or preventing cancer are provided, including administration to a patient in need thereof an effective amount of a compound described herein in combination with an additional method of treatment. In one embodiment, the additional method of treatment includes treatment with a chemotherapeutic agent. In one embodiment, the chemotherapeutic agent is one with which cancer treatment has not been found to be refractory. In another embodiment, the chemotherapeutic agent is one with which cancer treatment has been found to be refractory. The compound of the invention can be administered before, after, or at the same time as the chemotherapeutic agent.

In one embodiment, the additional method of treatment is radiation therapy. The compound of the invention can

<td>be administered</td><td>before,</td><td>after,</td><td>or at</td><td>same time as</td><td>the</td>

<td>radiation.</td><td></td><td></td><td></td><td></td><td></td>

<td>The compounds</td><td>of the</td><td>invention</td><td>too</td><td colspan="2">can be administered</td>

<td>to a patient</td><td colspan="3">that has been submitted or</td><td>will undergo</td><td>a</td>

Surgery as a treatment for cancer.

In a specific embodiment, the compound of the invention is administered simultaneously with the chemotherapeutic agent or with radiation therapy. In another specific embodiment, the chemotherapeutic agent or radiotherapy is administered before or after administration of the compound of the invention, in one aspect, at least one hour, five hours, 12 hours, one day, one week, one month, in other aspects several months (for example, up to three months), before or after the administration of a compound of the invention.

A chemotherapeutic agent can be administered through a series of sessions. Any or a combination of the chemotherapeutic agents listed herein or otherwise known in the art may be administered. With regard to radiation, any radiation therapy protocol can be used depending on the type of cancer to be treated. For example, but not by way of limitation, x-ray radiation can be administered; in particular, high-energy megavoltage (radiation greater than 1 MeV of energy) can be used for deep tumors, and electron beam and orthovoltage x-ray radiation can be used for skin cancers. Radioisotopes that emit gamma rays, such as radioactive isotopes of radio, cobalt and other elements, can also be administered.

In addition, methods of treating cancer with a compound of the invention are provided as an alternative to chemotherapy or radiation therapy where chemotherapy or radiation therapy has been demonstrated or may be too toxic, for example, results in side effects. unacceptable or unsustainable, for the subject to be treated. In addition, the methods of treating cancer with a compound of the invention are provided as an alternative to surgery in which surgery has proven or may prove unacceptable or unbearable for the subject being treated.

The compound of the invention can also be used in an in vitro or ex vivo manner, for example, for the treatment of certain types of cancer, including, not limited to leukemia and lymphomas, such treatment involves transplants of anthologous stem cells. This may involve a multi-stage process in which the anthology hematopoietic stem cells of the animal are collected and purged from all the cancer cells, the population of bone marrow cells of the remaining or remaining animal are then eradicated, through the administration of a high dose of a compound of the invention with or without the accompaniment of a high dose of radiation therapy, and the stem cell graft is again infused into the animal. Supportive care is provided, while

<td>the function</td><td>of the bone marrow</td><td>I know</td><td>restores and the</td><td>animal</td><td>I know</td>

<td>recover</td><td></td><td></td><td></td><td></td><td></td>

<td>Methods</td><td>for the treatment</td><td>of the</td><td>cancer include</td><td>further</td><td>the</td>

administration to a patient in need thereof an effective amount of a compound of the invention and another therapeutic agent that is an anti-cancer agent. Suitable anticancer agents include, but are not limited to, methotrexate, taxol, L-asparaginase, mercaptopurine, thioguanine, hydroxyurea, cytarabine, cyclophosphamide, ifosfamide, nitrosoureas, cisplatin, carboplatin, mitomycin, dacarbazine, procarbizine, nitrogen, topotecan , cytoxan, etoposide, 5-fluorouracil, BCNU, irinotecan, camptothecins, bleomycin, doxorubicin, idarubicin, daunorubicin, actinomycin D, dactinomycin, plicamycin, mitoxantrone, asparaginase, vinblastine, vincristine, vindesine, vinorelbine, paclitaxel and docetaxel. Other examples of chemotherapeutic agents include alkylating agents such as thiotepa and Cytoxan® cyclophosphamide; alkyl sulfonates such as busulfan, thiosulfan, improsulfan and piposulfan; aziridines such as benzodopa, carbocuone, meturedopa, and uredopa; ethyleneimines and methylamelamines including altretamine, triethylenemelamine, triethylene phosphoramide, triethylenediophosphoramide and trimethylolomelamine; TLK 286 (TELCYTA ™); acetogenins (especially bullatacin and bullatacinone); delta-9 tetrahydrocanabinol (dronabinol, Marinol®); beta-lapachone; lapachol; colchicine; betulinic acid; a camptothecin (including the synthetic analogue topotecan (HYCAMTIN®), CPT11 (irinotecan, CAMPTOSAR®), acetylcamptothecin, scopolectin, and 9-aminocamptothecin); biostatin; calistatin; CC-1065 (including its synthetic analogs adozelesin, carzelesin and bizelesin); podophyllotoxin; podophyllinic acid; teniposide; cryptophycin (particularly cryptophycin 1 and cryptophycin 8); dolastatin; duocarmycin (including synthetic analogs, KW-2189 and CB1TM1); eleutherobin; pancratistatin; sarcodictine; tub spongist; nitrogen mustards such as chlorambucil, chlornafazine, colophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine hydrochloride, melphalan, novembycin, phenesterine, prednimustine, trophophamide, and uracil mustard; triazines such as decarbazine; nitrosureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine and ranimnustine; epipodophyllins, such as etoposide, teniposide, topotecan, 9-aminocamptothecin, orcrisnatol camptothecin; bisphosphonates, such as clodronate; antibiotics such as enedin antibiotics (for example, caliqueamycin, especially gammall caliqueamycin and omegall caliqueamycin (see, for example, Agnew, Chem. Intl. Ed. Engl., 33: 183-186 (1994)) and η

anthracyclines such as anamycin, AD 32, alcarubicin, daunorubicin, dexrazoxane, DX-52-1, epirubicin, GPX-100, idarubicin, KRN5500, menogaryl, dynemycin, including dynemycin A, a speramycin, chromophoretophostetin-related chromosphorus and chromosphorus of enediine, clarcinomysins, actinomycin, autramycin, azaserine, bleomycins (for example, A2 and B2), cactinomycin, carabicin, caminomycin, carzinophilin, cromomycins, dactinomycin, detorubicin, 6diazo-5-oxo-L-norleucine, doxorubicin Adriamycin (including morpholino-doxorubicin, cianomorfolinodoxorrubicina, 2-pyrroline doxorubicin, liposomal doxorubicin and deoxydoxorubicin), esorubicin, marcellomycin, mitomycins such as mitomycin C, mycophenolic acid, nogalamycin, olivomycins, peplomycin, potfiromycin, puromycin, chelamicin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, and zorubicin; photodynamic therapies, such as vertoporfin (BPDMA), phthalocyanine, Pc4 photosensitizer and demethoxyhypocreline A (2BA-2-DMHA); folic acid analogs such as denopterin, pteropterin, trimetrexate; and purine analogs such as fludarabine, 6mercaptopurine, thiamiprine, and thioguanine; pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, cytosine arabinoside, dideoxyuridine, doxifluridine, enocitabine and floxuridine; androgens such as calusterone, dromostanolone propionate, epitiostanol, mepitiostane, and testolactone; anti-adrenals such as aminoglutethimide, mitotane and trilostane; folic acid booster such as folinic acid (leucovorin); aceglatone; antineoplastic anti-folate agents such as ALIMTA®, pemetrexed LY231514, dihydrofolate reductase inhibitors such as methotrexate and trimetrexate; antimetabolites such as 5fluorouracil (5-FU) and its prodrugs such as UFT, Sl and capecitabine, floxuridine, doxifluridine and ratitrexed; and thymidylate synthase inhibitors and glycinamide ribonucleotide formyltransferase inhibitors such as raltitrexed (TOMUDEX®, TDX); dihydropyrimidine dehydrogenase inhibitors such as enyluracil; aldophosphamide glycoside; aminolevulinic acid; amsacrine; bestrabucil; bisantrene; edatraxa to; defofamine; demecolcina; diazicuone; elformitin; eyliptinium acetate; an epothilone; ethtoglucid; gallium nitrate; hydroxyurea; lentinan; lonidainin; maitansinoids such as maitansin and ansamitocins; mitoguazone; mitoxantrone;

mopidanmol; nitraerin; pentostatin; fenamet; pyrarubicin; losoxantrone; 2-ethylhydrazide; procarbazine; PSK® polysaccharide complex (JHS Natural Producís, Eugene, Qreg.); razoxane; rhizoxin; sizofiran; spirogermanium; tenuazonic acid; triazicuone; 2,2 ', 2-trichlorotriethylamine; trichothecenes (especially T-2 toxin, verracurin A, roridine A and anguidine); urethane; vindesine (ELDISINE®, Fildesin®); dacarbazine; manomustine; mitobronitol; mitolactol; pipobroman; gacitosina; arabinoside (Ara-C); cyclophosphamide, thiotepa; taxoids and taxanes, for example, TAXOL® paclitaxel (Bristol-Myers Squibb Oncology, Princeton, NJ), ABRAXANE ™ free of cremfor, engineering nanoparticles of paclitaxel albumin engineering (American Pharmaceutical Partners, Schaumberg, 111.) and docetaxel TAXOTERE ® (Rhóne-Poulenc Rorer, Antony, France); chlorabucil; gemcitabine (Gemzar®); 6-thioguanine; mercaptopurine; platinum; platinum analogs or platinum-based analogs, such as cisplatin, oxaliplatin and carboplatin; vinblastine (Velban®); etoposide (VP-16);

ifosfamide; mitoxantrone; vincristine (Oncovin®); vinca alkaloids; vinorelbine (Navelbine®); velcade; revlimid; thalidomide; IMID3; lovastatin; verapamil; tapsigargin; 180 * '' I ♦ ís. w 1 *&gt; »methyl-4-phenylpyridinium; cell cycle inhibitors, such as staurosporine; novantrone; edatrexate; daunomycin, mtoxantrone; aminopterin; xeloda; ibandronate; topoisomerase inhibitor RFS 2000; difluoromethylornitine (DMFO); Vitamin D3 analogues, such as EB 1089, CB 1093 and KH 1060; retinoids such as retinoic acid; pharmaceutically acceptable salts, acids or derivatives of any of the foregoing; as well as combinations of two or more of the above such as CHOP, an abbreviation for a combined therapy of cyclophosphamide, doxorubicin, vincristine and prednisolone and FOLFOX, an abbreviation for a treatment regimen with oxaliplatin (ELOXANTIN ™) combined with 5-FU and leucovorin .

Anti-hormonal agents that act to regulate or inhibit hormonal action on tumors such as anti-estrogens and selective estrogen receptor modulators (SERMs), including, for example, tamoxifen (including tamoxifen Nolvadex®), raloxifene, megastrol, droloxifene , 4-hydroxy tamoxifen, trioxyphene, keoxifen, LY117018, onapristone, and Fareston® toremifene; aromatase inhibitors that inhibit the enzyme aromatase, which regulates the production of estrogens in the adrenal glands, such as, for example, 4 (5) imidazoles, aminoglutethimide, megestrol NEGASE® acetate, Aromasin® exemestane, formestanie, fadrozole, vorozole RIVISOR®, letrozole FEMARA®, and anastrozole ARIMIDEX®; and anti-androgens such as flutamide, bicalutamide, nilutamide, bicalutamide, leuprolide, and goserelin; as well as troxacitabine (a cystosine nucleoside 1,3-dioxolane analog); antisense oligonucleotides, particularly those that inhibit gene expression in signaling pathways involved in aberrant cell proliferation, such as, for example, PKC-alpha, Raf, Η-Ras, and epidermal growth factor receptor (EGF- R); vaccines such as gene therapy vaccines, for example, ALLOVECTIN® vaccine, LEUVECTIN® vaccine, and the VAXID® vaccine; Proleukin® rIL-2; 1 LURTOTECAN® topoisomerase inhibitor; ABARELIX® rmRH; and pharmaceutically acceptable salts, acids or derivatives of any of the foregoing.

The compounds of the present disclosure, or their pharmaceutically acceptable salts, may contain one or more asymmetric centers and therefore may give rise to enantiomers, diastereomers, and other stereoisomeric forms that can be defined, in terms of absolute stereochemistry, as (R) - or (S) - or, as (D) - or (L) - for amino acids. The present description is understood to include all these possible isomers, as well as their racemic and optically pure forms. Optically active (+) and (-), (R) - and (Sj-, or isomers (2?) - and (L) can be prepared using chiral syntons or chiral reagents, or resolved using conventional techniques, for example, chromatography and fractional crystallization. Conventional techniques for the preparation / isolation of individual enantiomers include chiral synthesis from a suitable optically pure precursor or resolution of the racemate (or the racemate of a salt or derivative) using, for example, chiral high pressure liquid chromatography (HPLC ). When the compounds described herein contain olefinic double bonds or other centers of geometric asymmetry, and unless otherwise specified, the compounds are intended to include both geometric isomers E and Z. Similarly, all tautomeric forms are also intended to be included.

A stereoisomer refers to a compound formed by the same atoms joined by the same bonds but having different three-dimensional structures, which are not interchangeable. The present description contemplates various stereoisomers and mixtures thereof and includes enantiomers, which refers to two stereoisomers whose molecules are mirror images of each other that are not superimposable.

A tautomer refers to a proton change from one atom of a molecule to another atom of the same molecule. The present description includes tautomers of any of said compounds.

The novel compounds

In one embodiment, the compounds having structure (I) are provided:

<img file="MX368258B_D0024.tif" />

.NR₆ OR ^R2^X \ (I) where:

Ri and R₂ they are independently selected from the group consisting of: H and a saturated or unsaturated residue having a linear, branched or non-aromatic cyclic skeleton containing from one to ten carbon atoms, and the carbon atoms are optionally substituted with: -OH, -I, -Br, Cl, -F, -CN, -CO₂H, -CHO, -COSH, or -NO₂; or R₂ and R₅ they fuse and form a ring;

R₃ and R₄ are independently selected from the group consisting of: H, R,

ArR-; or R3 and R₄ join together to form a ring;

Rs is selected from the group consisting of: H

R, ArR- and Ar;

or R₅ and R₂ they fuse and form a ring;

Re is selected from the group consisting of: H

R, and

ArR-;

R₇ and R₈ are independently selected from the group consisting of:

H, R, and

ArR-;

R9 is:

where

R is defined as a saturated or unsaturated residue having a branched, or non-aromatic, linear skeleton containing from one to ten carbon atoms, from zero to four nitrogen atoms, from zero to four oxygen atoms, and from zero to four sulfur atoms, and the carbon atoms are optionally substituted with: = 0, = S, OH, -OR10, -O₂CRi₀, -SH, -SRio, -SOCR10, -NH₂, -NHR₁₀, -N (R₀)₂, -NHCORiq, -NR₁₀COR₁₀, I, -Br, -Cl, -F, -CN, -CO₂H, -CO₂Ri₀, -CHO, -CORiq, -CONH₂, CONHR₁₀, -CON (R₁₀)₂, -COSH, -COSR10, -NO₂, -SO3H, -SOR10, SO₂R₁₀, where R₁₀ it is a linear, branched or cyclic, saturated or unsaturated alkyl group of one to ten carbons;

the ring formed by the union of R3 and R₄ it is a non-aromatic cyclic skeleton of three to seven members within the definition of R,

Y is defined as a residue selected from the group consisting of: a linear, saturated or unsaturated alkyl group of one to six carbons, optionally substituted with R, ArR-, or X; Y,

X is defined as a residue selected from the group consisting of: -OH, -OR, = 0, = S, -O₂CR, -SH, -SR, -SOCR, -NH₂, NHR, —N (R)₂, -NHCOR, -NRCOR, -I, -Br, -Cl, —F, -CN, -CO₂H, CO₂R, -CHO, -COR, -CONH₂, -CONHR, -CON (R)₂, -COSH, -COSR, NO₂, -SO₃H, -SOR, and —SO₂R;

R14 is selected from the group consisting of optionally substituted alkyl, optionally substituted alkylamino, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heterocyclyl and optionally substituted heteroaryls, COR₂₄, CSR₂₄, -OR₂4, and -NHR24, wherein each R24 is independently alkyl optionally substituted with halogen, -OH or -SH;

or a stereoisomer, prodrug or pharmaceutically acceptable salt thereof.

In one embodiment, Ar is an aromatic ring selected from the group consisting of: phenyl, naphthyl, anthracil, pyrrolyl.

In one embodiment, compounds having the following structure (the) are provided

<img file="MX368258B_D0025.tif" />

where:

Ri4 is selected from the group consisting of optionally substituted alkyl, optionally substituted alkylamino, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heterocyclyl, optionally substituted heteroaryl, COR₂4, CSR₂4, -OR24, and -NHR₂4 &lt;where each R₂4 it is, independently, alkyl optionally substituted with halogen, -OH or -SH;

Ris is selected from the group consisting of optionally substituted alkyl, optionally substituted alkylamino, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heterocyclyl and optionally substituted heteroaryl;

Ri₆ is selected from the group consisting of H and Ci6 alkyl &lt;*

R17 is selected from the group consisting of H and C6 alkyl / acyl

Laughs and R30 are independently selected from the group consisting of H, C1-6 alkyl and -SH, with the proviso that Ris and R30 cannot be both H;

R19, ​​R20, R21 and R22 are independently H and Ci_ alkyl₆, at least one of R19 and R₂or is H; or R₂o and R21 form a double bond, R19 is H, and R₂₂ is H or Ci_ alkyl₆; Y

R₂₃ it is selected from the group consisting of H and Cie alkyl; or a stereoisomer, prodrug or pharmaceutically acceptable salt thereof.

In a further embodiment, each optionally substituted alkyl, optionally substituted alkylamino, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heterocyclic and optionally substituted heteroaryl is, independently, optionally substituted with = 0, = S, -OH, -OR₂₄, -O₂CR₂₄, -SH, -SR₂₄, -socr₂₄, -nh₂, -N₃, -NHR₂₄, -N (R₂₄)₂, -NHCOR₂₄, NR₂₄COR₂₄, -I, -Br, -Cl, -F, -CN, -CO₂H, -CO₂R₂₄, -CHO, -COR₂₄-conh₂, -conhr₂₄, -CON (R₂₄)₂, -cosh, -cosr₂₄, -do not₂-so₃h, SOR₂₄ bear₂R₂₄ where each R₂₄ it is, independently, alkyl optionally substituted with halogen, -OH or -SH.

In another additional embodiment, each optionally substituted aryl and optionally substituted heteroaryl is independently selected from the group consisting of optionally substituted phenyl, optionally substituted naphthyl, optionally substituted anthracyl, optionally substituted phenanthryl, optionally substituted furyl, optionally substituted pyrrolyl, optionally substituted thiophenyl , optionally substituted benzofuryl, optionally substituted benzothiophenyl, optionally substituted quinolinyl, optionally substituted isoquinolinyl, optionally substituted imidazolyl, optionally substituted thiazolyl, optionally substituted oxazolyl, and optionally substituted pyridinyl.

In another additional mode, Ri₅ is selected from one of the following structures (II), (III), (IV), (V):

<img file="MX368258B_D0026.tif" />

(ID

<img file="MX368258B_D0027.tif" />

(III) (IV) (V) where:

Q is CR25 or N;

Z is C (R₂5)₂, NR25, s, OO;

every R₂5 is independently selected from the group consisting of H, -OH, -R₂₄, -OR₂₄, -O₂CR₂₄, -SH, -SR₂₄, -SOCR₂₄, NH₂, -n₃, -NHR₂₄, -N (R₂₄)₂, -nhcor₂₄, -nr₂₄cor₂₄, -R₂₄NH₂, -I, Br, -Cl, -F, -CN, -CO₂H, -CO₂R₂₄, -CHO, -COR₂₄, -CONH₂, CONHR₂₄, -CON (R₂₄)₂, -COSH, -COSR₂₄, -do not₂-so₃h, -SOR₂₄ bear₂R₂₄, where R₂₄ it is, independently, alkyl optionally substituted with halogen, -OH or -SH.

In another additional mode, R15 is selected from the group consisting of:

<img file="MX368258B_D0028.tif" />

<img file="MX368258B_D0029.tif" />

where each R₂s is independently selected from the group consisting of H, -OH, -R₂₄, -OR₂₄, -O₂CR₂₄, -SH, -SR₂₄,

-SOCR₂₄, -NH₂, -N₃, -NHR₂₄, -N (R₂₄) 2, -nhcor₂₄, -nr₂₄cor₂₄, R₂₄NH₂, -I, -Br, -Cl, -F, -CN, -CO₂H, -CO₂R₂₄, -CHO, -COR₂₄, CONH₂, -CONHR₂₄, -CON (R₂₄)₂, -cose, -cosr₂₄, -do not₂-so₃h, -sor₂₄bear₂R₂₄, where each R₂₄ it is, independently, alkyl optionally substituted with halogen, -OH or -SE.

In another additional mode, R15 is selected from the group consisting of:

<img file="MX368258B_D0030.tif" />

<img file="MX368258B_D0031.tif" />

<img file="MX368258B_D0032.tif" />

<img file="MX368258B_D0033.tif" />

<img file="MX368258B_D0034.tif" />

<img file="MX368258B_D0035.tif" />

<img file="MX368258B_D0036.tif" />

In another additional mode, R_X5 it is:

<img file="MX368258B_D0037.tif" />

In another additional mode, R_í6, R_i7, R₁₈, and R₃₀ They are each methyl.

In another additional mode, R₁₆, is H, R_i7 it's methyl, R₁₈ it is methyl and R₃₀ It is methyl.

It is understood that any modality of the compounds of structure (la), as set forth above, and any specific substituent set forth herein for a group R₁₄, R₁₅, R₁₆, R₁₇, Ri₈, R₁₉, R₂₀ and R₃or in the compounds of structure (la), as set forth above, they can be combined independently with other modalities and / or substituents of compounds of structure (I) to form modalities of the present specification not specifically set forth above. In addition, in the event that a list of substituents is listed for any R₁₄, R_X5, R_X6, R_X7, Rig, R19, R20 and R30 in a particular embodiment and / or claim, it is understood that each individual substituent may be removed from the particular modality and / or claim, and that the remaining list of substituents shall be deemed to be within the scope of this description.

In one embodiment, compounds that have the following structure (Ib) are provided:

<img file="MX368258B_D0038.tif" />

(Ib) where:

R26 is selected from the group consisting of optionally substituted alkyl, optionally substituted alkylamino, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heterocyclyl and optionally substituted heteroaryl;

R27 is selected from the group consisting of optionally substituted alkyl, optionally substituted alkylamino, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heterocyclyl and optionally substituted heteroaryl;

Ri6 is selected from the group consisting of

H and Ci_ alkyl

R17 is selected from the group consisting of H and CiRi alkyl₈ is selected from the group consisting of Ci_ alkyl₆ and 93

SH, or a stereoisomer, prodrug or pharmaceutically acceptable salt thereof.

In a further embodiment, each optionally substituted alkyl, optionally substituted alkylamino, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heterocyclyl and optionally substituted heteroaryl is, independently, optionally substituted with = 0, = S, -OH, -OR₂₈&lt;-O₂CR₂₈, -SH, -SR₂₈, -SOCR₂₈, -NH₂, -N₃, -NHR₂₈, -N (R₂₈)₂, -NHCOR₂₈, NR₂₈COR₂₈, -I, -Br, -Cl, -F, -CN, -CO₂H, -CO₂R₂₈, -CHO, -COR₂₈-conh₂, -CONHR₂₈, -CON (R₂₈)₂, -cosh, -cosr₂₈, -do not₂-so₃h, SOR₂₈ bear₂R₂₈, where each R₂₈ it is, independently, alkyl optionally substituted with halogen, -OH or -SH.

In another additional embodiment, each optionally substituted aryl and optionally substituted heteroaryl is independently selected from the group consisting of optionally substituted phenyl, optionally substituted naphthyl, optionally substituted anthracyl, optionally substituted phenanthryl, optionally substituted furyl, optionally substituted pyrrolyl, optionally substituted thiophenyl , optionally substituted benzofuryl, optionally substituted benzothiophenyl, optionally substituted quinolinyl, optionally substituted isoquinolinyl, optionally substituted imidazolyl, optionally substituted thiazolyl, optionally substituted oxazolyl, and optionally substituted pyridinyl.

In another additional mode, R₂? it is selected from one of the following structures (II), (III), (IV), (V):

that

I

Q. Ά Q '

<img file="MX368258B_D0039.tif" />

(III)

<img file="MX368258B_D0040.tif" />

(IV)

<img file="MX368258B_D0041.tif" />

(V) where:

Q is CR29 or N;

Z is C (R29) 2, NR29, S, or O, each R29 is independently selected from the group consisting of H, -OH, -OR₂₈, -O₂CR₂₈, -SH, -SR₂₈, -SOCR₂₈, -NH₂,

N₃, -NHR₂₈, -N (R₂₈)₂, -NHCOR₂₈, -NR₂₈COR₂₈, -I, -Br, -Cl, -F, CN, -co₂h, -co₂r₂₈, -cho, -cor₂₈-conh₂, -conhr₂₈, -CON (R₂₈)₂, COSH, -COSR₂₈z ~ NO₂, -SO₃H, -SOR₂₈ bear₂R₂₈, where each

R₂₈ it is, independently, alkyl optionally substituted with halogen, -OH or -SH.

In another additional mode, R₂7 It is selected from the group consisting of:

<img file="MX368258B_D0042.tif" />

<img file="MX368258B_D0043.tif" />

<img file="MX368258B_D0044.tif" />

In another additional mode, R₂₇ it is:

<img file="MX368258B_D0045.tif" />

Ri₈ are each

In another additional modality methyl.

In another additional mode, Ri₆ it's H, R₁₇ is methyl, and R₁₈ It is methyl.

It is understood that any modality of the compounds of structure (Ib) as set forth above, and any specific substituent set forth herein for a group R₂Mr₂6z Ri6z Ri_7/ Risz Ris and R20 in the compounds of structure (Ib), as set forth above, may be independently combined with other modalities and / or substituents of structure compounds to form modalities of the present specification not specifically set forth above. In addition, in the event that a list of substituents is listed for any R₂₅, R₂6, Ri6z

R₁₇, Ri8 &lt;Ris and R20 particular in a particular embodiment and / or claim, it is understood that each individual substituent can be removed from the particular claim mode, and that the remaining list of substituents will be considered to be within the scope of the present description.

In one embodiment, the invention provides a method of manufacturing a compound having structure (I), (la) or (Ib).

Conjugates comprising new compounds

Compounds having the structure (I), (la) or (Ib) can be used to form conjugates, for example, antibody-drug conjugates (ADCs). Accordingly, in one embodiment of the present description, conjugate compositions are provided having the following structure:

(T) - (L) - (D) (VI) where (T) is a target residue, (L) is an optional linker, and (D) is a compound having structure (I), (la) or (Ib), below. In one embodiment, (T) is an antibody. Accordingly, in one embodiment, antibody-drug conjugates (ADCs) comprising compounds (D) having structure (I), (la) or (Ib) are provided.

As will be appreciated by the person skilled in the art, a wide variety of means are available to covalently link (T) - (L) - (D). Any known method can be used to link conjugated components. Any technology to generate links can be used to link (T) to (D). In addition, (T), (L), and (D) may be modified in any suitable manner, as recognized by the person with reasonable knowledge in the art, in order to facilitate conjugate formation. Target residue (T)

The target residue (T) of the compositions object of the present application includes within its scope any unit of a (T) that reactively binds or associates with a receptor, antigen or other receptive residue associated with a population of given target cells . A (T) is a molecule that binds to, complexes with, or reacts with a residue from a population of cells that are targeted. In one aspect, (T) acts to release the drug (D) to the particular target cell population with which (T) reacts. Such (T) s include, but are not limited to, large molecular weight proteins, such as, for example, full length antibodies, antibody fragments, smaller molecular weight proteins, polypeptide or peptides, lectins, glycoproteins, not peptides, vitamins, nutrient transport molecules (such as, but not limited to, transferrin), or any other cell binding molecule or substance.

A (T) can form a link to a linker unit (L) or a drug (D). A (T) can form a bond with a unit (L) through a heteroatom of (T). Heteroatoms that may be present in a (T) include sulfur (in one embodiment, from a sulfhydryl group of one (T)), oxygen (in one embodiment, from a carbonyl, carboxy or hydroxyl group of one (T)) and nitrogen (in one embodiment, from a primary or secondary amino group of a (T)). These heteroatoms may be present in (T) in the natural state of (T), for example an antibody of natural origin, or may be introduced in (T) through chemical modification.

In one embodiment, a (T) has a sulfhydryl group and (T) binds to (L) through a sulfur atom of the sulfhydryl group. In another embodiment, (T) has one or more Usina residues that can be chemically modified to introduce one or more sulfhydryl groups. The (T) binds to the unit (L) through the sulfhydryl group. Reagents that can be used to modify lysines include, but are not limited to, N-succinimidyl S-acetyl thioacetate (SATA) and 2-iminothiolane hydrochloride (Traut reagent).

In another embodiment, (L) may have one or more carbohydrate groups that can be chemically modified to have one or more sulfhydryl groups. The (T) binds to (L) through a sulfur atom of the sulfhydryl group. In yet

100 Another embodiment, (T) may have one or more carbohydrate groups that can be oxidized to provide an aldehyde group (-CHO) (see, for example, Laguzza et al, 1989, J. Med. Chem. 32 ( 3): 548-55). The corresponding aldehyde can form a bond with a reactive site in a portion of a (L). Reactive sites that can react with a carbonyl group in a (T) include, but are not limited to, hydrazine and hydroxylamine. Other protocols for protein modification for the binding or association of (D) are described in Coligati et al., Current Protocols in Protein Science, vol 2, John Wiley &amp; Sons (2002), incorporated herein by reference.

The (T) may include, for example, a protein, polypeptide or peptide including, but not limited to, transferrin factors, epidermal growth (EGF), bombesin, gastrin, gastrin releasing peptide, platelet derived growth factor , IL-2, IL-6, transforming growth factor (TGF), such as TGF-α or TGF-β, vaccinia virus growth factor (VGF), insulin growth factors and insulin-like I and II , lectins and apoprotein of low density lipoproteins. The (T) may also include an antibody, such as polyclonal antibodies or monoclonal antibodies. The

101 antibody may be directed to a particular antigenic determinant, including, for example, a cancer cell antigen, a viral antigen, a microbial antigen, a protein, a peptide, a carbohydrate, a chemical, nucleic acid, or fragments of the same. Methods for producing polyclonal antibodies are known in the art. A monoclonal antibody (mAb) for an antigen of interest can be prepared using any technique known in the art. These include, but are not limited to, the hybridoma technique originally described by Kohler and Milstein (1975 Nature 256, 495-497), the human B-cell hybridoma technique (Kozbor et ai, 1983, Immunology Today 4: 72) , and the EBV hybridoma technique (Colé et al., 1985, Monoclonal Antibodies and Cancer Therapy, Alan R, Liss, Inc., pp. 77-96). The Antibody Method

<td>Lymphocyte</td><td>Selected (</td><td colspan="2">SLAM) (Babcook, JS, et</td><td>to the,</td><td>A novel</td>

<td>strategy</td><td>for generating</td><td>monoclonal</td><td>antibodies</td><td>desde</td><td>single,</td>

<td>isolated</td><td>lymphocytes</td><td>producing</td><td>antibodies</td><td>of</td><td>defined</td>

<td colspan="2">specificities Proc Nati</td><td>Acad Sci U</td><td>SA, 1996.</td><td> 93</td><td>(15): p.</td>

7843-8) and (McLean GR, Olsen OA, Watt IN, Ratha naswami P, Leslie KB, Babcook JS, Schrader JW. Recognition of human cytomegalovirus by human primary immunoglobulins identifies an innate foundation to an adaptive immune response.

102

Immunol 2005 Apr 15; 174 (8): 4768-78.) Such antibodies may be of any immunoglobulin class including IgG, IgM, IgE, IgA, and IgD and any subclass thereof. The hybridomas that produce the mAbs for use in this invention can be cultured in vitro or in vivo.

The monoclonal antibody can be, for example, a human monoclonal antibody, a humanized monoclonal antibody, an antibody fragment, or a chimeric antibody (for example, a mouse-human antibody). Human monoclonal antibodies can be prepared by any of the numerous techniques known in the art (eg, Teng et al., 1983, Proc. Nati. Acad. Sci. USA 80: 7308-7312; Kozbor et al., 1983, Immunology Today 4: 72-79; and Olsson et al., 1982, Meth. Enzymol. 92: 3-16). See also, Huse et al., 1989, Science 246: 1275-1281 and McLean et al. J Immunol. 2005 Apr 15; 174 (8): 4768-78.

The antibody can also be a bispecific antibody. Methods for preparing bispecific antibodies are known in the art. Traditional production of full-length bispecific antibodies is based on the coexpression of two pairs of immunoglobulin light chain heavy chains, where the two chains have different specificities (see, for example, Milstein

103 et al., 1983, Nature 305: 537-539; International Publication No. WO 93/08829, Traunecker et al., 1991, EMBO J. 10: 36553659).

According to a different approach, the variable domains of antibodies with the desired binding specificities (antibody-antigen combination sites) are fused to immunoglobulin constant domain sequences. The fusion is preferably with a constant domain of the immunoglobulin heavy chain, comprising at least part of the hinge, regions C_Htwo and C_H3 · It is preferred to have the first heavy chain constant region (C_Hi) which contains the site necessary for light chain binding, present in at least one of the fusions. Nucleic acids with sequences encoding immunoglobulin heavy chain fusions and, if desired, the immunoglobulin light chain, are inserted into separate expression vectors and co-transfected into a suitable host organism. This provides flexibility in adjusting the mutual proportions of the three polypeptide fragments in modalities when the unequal proportions of the three polypeptide chains used in the construction provide the optimum yields. However, it is possible to insert the coding sequences for two or all three polypeptide chains

104 in an expression vector when the expression of at least two polypeptide chains in equal proportions results in high yields or when the proportions are not of particular significance.

For example, bispecific antibodies may have a hybrid immunoglobulin heavy chain with a first binding specificity in one arm, and a heavy chain-hybrid immunoglobulin light chain pair (which provides a second binding specificity) in the other arm. This asymmetric structure facilitates the separation of the desired bispecific compound from combinations of unwanted immunoglobulin chains, since the presence of an immunoglobulin light chain in only one half of the bispecific molecule provides an easy way of separation (International Publication No. WO 94 / 04690) which is incorporated herein by reference in its entirety.

For more details to generate bispecific antibodies see, for example, Suresh et al., 1986, Methods in Enzymology 121: 210; Rodrigues et al., 1993, J. Immunology 151: 6954-6961; Carter et al., 1992, Bio / Technology 10: 163167; Carter et al., 1995, J. Hematotherapy 4: 463-470; Merchant et al., 1998, Nature Biotechnology 16: 677-681. Using such techniques, bispecific antibodies are

105 they can prepare for use in the treatment or prevention of diseases as defined herein.

Bifunctional antibodies are also described in European patent EPA 0 105 360. As described in this reference, hybrid or bifunctional antibodies can be derived either biologically, that is, by cell fusion techniques, or chemically, especially with crosslinking agents or disulfide bridge formation reagents, and may comprise whole antibodies or fragments thereof. The methods for obtaining such hybrid antibodies are described, for example, in International Publication WO 83/03679, and European Patent Publication No. EPA 0 217 577, both of which are incorporated herein by reference.

The antibody may also be a functionally active fragment, derived or analogous from an antibody that immunospecifically binds to a target antigen (eg, a cancer antigen, a viral antigen, a microbial antigen, or other antibodies bound to cells or matrix) . In this sense, functionally active means that the fragment, derivative or analog is capable of recognizing the same antigen as the antibody, from which the fragment, derivative or analog is derived, recognized. Specifically in

106 An exemplary embodiment of the antigenicity of the idiotype of the immunoglobulin molecule can be enhanced by deleting framework and CDR sequences that are C-terminal to the CDR sequence that specifically recognizes the antigen. To determine which CDR sequences bind to the antigen, synthetic peptides containing the CDR sequences can be used in antigen binding assays by any binding assay method known in the art (e.g., the BIA core assay) (See, for example, Kabat et al., 1991, Sequences of Proteins of Immunological Interest, Fifth Edition, National Institute of Health, Bethesda, Md .; Kabat et al., 1980, J. Immunology 125 (3): 961- 969).

Other useful antibodies include antibody fragments such as, but not limited to, F (ab ') fragments.₂, Fab, Fab 'fragments, Fv fragments and heavy chain and light chain antibody dimers, or any minimum fragment thereof such as Fvs or single chain antibodies (SCAs) (for example, as described in US Pat. No. 4,946,778; Bird, 1988, Science 242: 423-42; Huston et al., 1988, Proc. Nati. Acad. Sci. USA 85: 5879-5883; and Ward et al., 1989, Nature 334: 544-54 ).

Recombinant antibodies, such as antibodies

107 Chimeric and humanized monoclonal, comprising both human and non-human portions, which can be made using standard recombinant DNA techniques, can also be used. (See, for example, U.S. Patent No. 4,816,567; and U.S. Patent No. 4,816,397). Humanized antibodies are antibody molecules of non-human species that have one or more complementarity determining regions (CDRs) of non-human species and a framework region of a human immunoglobulin molecule (see, for example, US Patent No. 5,585,089). Chimeric and humanized monoclonal antibodies can be produced by recombinant DNA techniques known in the art, for example using methods described in International Publication No. WO 87/02671; European Patent Publication No. 0 184

<td> 187;</td><td>Publication</td><td>from</td><td>Patent</td><td>European</td><td>Do not.</td><td> 0</td><td> 171</td>

<td> 4 96;</td><td>Publication</td><td>from</td><td>Patent</td><td>European</td><td>Do not.</td><td> 0</td><td> 173</td>

<td> 4 94;</td><td>Publication</td><td colspan="2">International No.</td><td colspan="2">WO 86/01533;</td><td>Patent</td><td>from</td>

U.S. No, 4,816,567; European Patent Publication No. 012 023; Berter et al., 1988, Science 240: 1041-1043; Liu et al., 1987, Proc. Nati Acad. Sci. USA 84: 3439-3443; Liu et al., 1987, J. Immunol. 139: 3521-3526; Sun et al., 1987, Proc. Nati Acad. Sci. USA „84: 214-218; Nishimura et al., 1987,

108

Cancer. Res. 47: 999-1005; Wood et al., 1985, Nature 314: 446449; Shaw et al., 1988, J. Nati. Cancer Inst. 80: 1553-1559; Morrison, 1985, Science 229: 1202-1207; Oi et al., 1986, BioTechniques 4: 214; U.S. Patent No. 5,225,539; Jones et al., 1986, Nature 321: 552-525; Verhoeyan et al., 1988, Science 239: 1534; and Beidler et al., 1988, J. Immunol. 141: 4053-4060.

Fully human antibodies can be used. Human antibodies can be prepared, for example, using transgenic mice that are incapable of expressing endogenous immunoglobulin heavy and light chain genes, but which can express human heavy and light chain genes. Transgenic mice are immunized in the normal manner with a selected antigen, for example, all or a portion of a polypeptide of the invention. Monoclonal antibodies directed against the antigen can be obtained using conventional hybridoma technology. Human immunoglobulin transgenes housed by transgenic mice are rearranged during B cell differentiation, and subsequently subjected to class change and somatic mutation. Therefore, using such a technique, it is possible to produce therapeutically useful IgG, IgA, IgM and IgE antibodies. For an overview of this

109 technology for the production of human antibodies, see Lonberg and Huszar (1995, Int. Rev. Immunol. 13: 65-93). For a detailed discussion of this technology for producing human antibodies and human monoclonal antibodies and protocols for producing such antibodies, see, for example, United States Patents No. 5,625,126; 5,633,425; 5,569,825; 5,661,016; and 5,545,806.

Human antibodies that recognize a selected epitope can also be generated using a technique called guided selection. In this approach a selected non-human monoclonal antibody, for example, a mouse antibody, is used to guide the selection of a completely human antibody that recognizes the same epitope. (See, for example, Jespers et al., 1994, Biotechnology 12: 899-903). Human antibodies can also be produced using various techniques known in the art, including phage display libraries (see, for example, Hoogenboom and Winter, 1991, J. Mol. Biol. 227: 381; Marks et al., 1991, J. Mol. Biol. 222: 581; Quan and Carter, 2002, The rise of monoclonal antibodies as therapeutics, in Anti-IgE and Allergic Disease, Jardieu, PM and Fick Jr., R. B, eds., Marcel Dekker, New York, NY, Chapter 20, pp. 427469).

110

In other embodiments, the antibody is a fusion protein of an antibody, or a functionally active fragment thereof. For example, an antibody can be fused through a covalent bond (for example, a peptide bond), either at the N-terminal or C-terminal ends to an amino acid sequence of another protein (or portion thereof, such as at least a 10, 20 or 50 amino acid portion of the protein) that is not the antibody.

The antibodies also include analogs and derivatives that are modified, that is, either by covalent binding of any type of molecule provided that said covalent binding allows the antibody to retain its antigen binding immunospecificity. For example, but not by way of limitation, antibody derivatives and analogs include those that have been further modified, for example, by glycosylation, acetylation, pegylation, phosphorylation, amidation, derivatization by known protecting / blocking groups, proteolytic cleavage, binding to a unit of cellular antibody or other protein, etc. Any of the numerous chemical modifications can be carried out by known techniques, including, but not limited to specific chemical cleavage, acetylation, formylation, metabolic synthesis in the presence of

111 tunicamycin, etc. In addition, the analog or derivative may contain one or more unnatural amino acids.

Antibodies may have modifications (eg, substitutions, deletions or additions) in amino acid residues that interact with Fe receptors. In particular, antibodies include antibodies that have modifications in amino acid residues identified as being involved in the interaction between the domain. anti-Fc and the FcRn receptor (see, for example, International Publication No. WO 97/34631, which is incorporated herein by reference in its entirety). Immunospecific antibodies to a target antigen can be obtained commercially or from another source or produced by any method known to a person skilled in the art such as, for example, chemical synthesis or recombinant expression techniques. The nucleotide sequence encoding immunospecific antibodies to a cancer cell antigen can be obtained, for example, from the GenBank database or a similar database, literature publications, or by cloning and sequencing of routine.

Examples of antibodies available for the treatment of cancer include, but are not limited to, humanized monoclonal antibody against HER2, HERCEPTIN® (trastuzumab;

112

Genentech); RITUXAN® (rituximab; Genentech) which is a chimeric anti-CD20 monoclonal antibody for the treatment of patients with non-Hodgkin lymphoma; OvaRex (AltaRex Corporation, MA), which is a murine antibody for the treatment of ovarian cancer; Panorex (Glaxo Wellcome, NC), which is a murine IgG2a antibody for the treatment of colorectal cancer; Cetuximab Erbitux (Imclone Systems Inc., NY) which is an anti-EGFR chimeric IgG antibody for the treatment of cancers positive for epidermal growth factor, such as head and neck cancer; Vitaxin (Medlmmune, Inc., MD) which is a humanized antibody for the treatment of sarcoma; Campath I / H (LeukoSite, MA) which is a humanized IgGl antibody for the treatment of chronic lymphocytic leukemia (CLL); Smart MI95 (Protein Design Labs, Inc., CA) which is a humanized anti-CD33 IgG antibody for the treatment of acute myeloid leukemia (AML); LymphoCide (Immunomedics, Inc., NJ) which is a humanized anti-CD22 IgG antibody for the treatment of non-Hodgkin lymphoma; Smart ID10 (Protein Design Labs, Inc., CA) which is a humanized anti-HLA-DR antibody for the treatment of non-Hodgkin lymphoma; Oncolym (Techniclone, Inc., CA) which is a radiolabeled murine anti-HLA-DRIO antibody for the treatment of non-Hodgkin lymphoma;

113

Allomune (Transο Transplant, CA) which is a humanized anti-CD2 mAb for the treatment of Hodgkin's disease or non-Hodgkin's lymphoma; Avastin (Genentech, Inc., CA) which is a humanized anti-VEGF antibody for the treatment of lung and colorectal cancer; Epratuzamab (Immunomedics, Inc., NJ and Amgen, CA) which is an anti-CD22 antibody for the treatment of non-Hodgkin lymphoma; and CEAcide (Immunomedics, NJ) which is a humanized anti-CEA antibody for the treatment of colorectal cancer.

Other antibodies useful in the treatment of cancer include, but are not limited to, antibodies against the following antigens (exemplary cancers are indicated in brackets): CA125 (ovary), CA15-3 (carcinomas), CA19-9 (carcinomas), L6 (carcinomas), Lewis Y (carcinomas), Lewis X (carcinomas), fetoprotein alfa (carcinomas), CA 242 (colorectal), alkaline phosphatase of the placenta (carcinomas), antigen of the specific membrane of the prostate (prostate), prostatic acid phosphatase (prostate), epidermal growth factor (carcinomas), MAGE-1 (carcinomas), MAGE-2 (carcinomas), MAGE- 3 (carcinomas), MAGE4 (carcinomas), anti transferrin receptor (carcinomas), p97 (melanoma), MUC1-KLH (breast cancer), CEA (colorectal), gplOO (melanoma), MARTI (melanoma), specific antigen

114 Prosthetic (PSA) (prostate), IL-2 receptor (leukemia and T-cell lymphomas), CD20 (non-Hodgkin lymphoma), CD52 (leukemia), CD33 (leukemia), CD22 (lymphoma), human chorionic gonadotropin (carcinoma) , CDS38 (multiple myeloma), CD40 (lymphoma), mucin (carcinomas), P21 (carcinomas), MPG (melanoma), and Neu oncogenic product (carcinomas). Some specific, useful antibodies include, but are not limited to, BR96 mAb (Trail et al., 1993, Science 261: 212-215), BR64 (Trail et al., 1997, Cancer Research 57: 100-105), mAbs against the CD40 antigen, such as mAb S2C6 (Francisco et al., 2000, Cancer Res. 60: 3225-3231) and chimeric and humanized variants thereof, mAbs against the cD33 antigen; mAbs against the EphA2 antigen; mAbs against the CD70 antigen, such as mAb 1F6 and mAb 2F2 and chimeric and humanized variants thereof, and mAbs against the CD30 antigen, such as AC10 (Bowen et al., 1993, J. Immunol 151: 5896-5906; Wahl et al., 2002, Cancer Res. 62 (13): 3736-42) and chimeric and humanized variants thereof. Many other internalization antibodies that bind to tumor-associated antigens can be used and have been reviewed in the literature (see, for example, Franke et al., 2000, Cancer Biother. Radiopharm. 15: 459 76; Murray, 2000, Semin. Oncol. 27:64 70; Breitling et al., Recombinant Antibodies, John Wiley, and Sons, New York,

115

1998) .

The antibody can also be an antibody that binds to an antigen that is present in a target cell or a population of target cells. For example, transmembrane polypeptides and other markers can be specifically expressed on the surface of one or more particular types of target cells (for example, a cancer cell) compared to one or more normal (for example, one (s) cell ( s) not cancerous). Often, such markers are expressed more abundantly on the surface of the target cells, or have a higher immunogenicity, compared to those on the surface of normal cells. The identification of such cell surface antigen polypeptides has resulted in the ability to specifically target cells for destruction through antibody-based therapies. Therefore, in some embodiments, antibodies include, but are not limited to, antibodies against tumor associated antigens (TAA). Such tumor associated antigens are known in the art, and can be prepared for use in the generation of antibodies using methods and information that are well known in the art.

See also EP2552957, WO / 2012/116453, WO / 2012/032080. See

116 also Zybody ™, http://www.zyngenia.com/technology.html. See also human heavy chain only antibody technology, http://www.crescendobiologics.com/. See also W02010001251, yeast based platform human yeast based antibody http://www.adimab.com/science-andtechnology/technology-overview/, mAbLogix ™ platform http://www.dna.com/technology, discovery platform monoclonal http://www.igenica.com/technology/, WO2009 / 157771, EP2560993, W02013004842, WO2012166560.

Bonding Residue (L)

The compositions object of the present optionally also include a linker residue (L). (L) is a bifunctional compound that can be used to bind a (D) and a (T) to form a conjugate composition, TLD. Such conjugates allow selective delivery of drugs to target cells (eg, tumor cells). The (L) s include a divalent substituent such as alkyldiyl, an aryldiyl, a heteroaryldiyl, residues such as: (CR₂) _n0 (CR₂) n ~~ i repeating units of alkyloxy (for example, polyethylene, PEG, polymethyloxy) and alkylamino (eg, polyethyleneamine, Jeffamine ™); and ester of diacid and amides, including succinate, succinamide, diglycolate, malonate, and caproamide.

117

The compositions object of the present can be prepared using a unit (L) having a reactive site for binding to (D) and (T). In some embodiments, (L) has a reactive site that has an electrophilic group that is reactive to a nucleophilic group present in (T). Useful nucleophilic groups on (T) include, but are not limited to sulfhydryl, hydroxyl and amino groups. The nucleophile group heteroatom of (T) is reactive to an electrophilic group in (L) and forms a covale bond to (L). Useful electrophilic groups include, but are not limited to maleimide and haloacetamide groups. The nucleophile group in (T) provides a convenient site for binding to (L).

In another embodiment, (L) has a reactive site that has a nucleophilic group that is reactive to an electrophilic group present in (T). Useful electrophilic groups on (T) include, but are not limited to, carbonyl aldehyde and ketone groups. The heteroatom of a nucleophilic group of (L) can react with an electrophilic group in (T) and form a covalent bond to (T). Useful nucleophilic groups on (L) include, but are not limited to, hydrazide, oxime, amino, hydrazine, thiosemicarbazone, hydrazine carboxylate, and arylhydrazide. The electrophilic group in (T) provides a convenient site for binding to (L).

118

The carboxylic acid functional groups and chloroformate functional groups are also useful reactive sites for (L), since they can react with amino groups of one (D) to form an amide bond. Also useful as a reactive site is the carbonate functional group in (L), such as, but not limited to p-nitrophenyl carbonate, which can react with an amino group of a (D) to form a carbamate bond.

It will be appreciated that any linker residues taught in the prior art, and in particular those that have been taught for use in the context of drug delivery, can be used in the present invention. Without limiting the scope of the above statement, in one embodiment, (L) comprises a linker residue disclosed in WO 2012/113847. In another embodiment, (L) comprises a linker residue disclosed in US 8,288,352. In another embodiment, (L) comprises a linker residue disclosed in US 5,028,697. In another embodiment, (L) comprises a linker residue disclosed in US 5,006,652.

In another embodiment, (L) comprises a linker residue disclosed in US 5,094,849. In another embodiment, (L) comprises a linker residue disclosed in US 5,053,394. In another embodiment, (L) comprises a residue

119 linker disclosed in US 5,122,368. In another embodiment, (L) comprises a linker residue disclosed in US 5,387,578. In another embodiment, (L) comprises a linker residue disclosed in US 5,547,667. In another embodiment, (L) comprises a linker residue disclosed in US 5,622,929. In another embodiment, (L) comprises a linker residue disclosed in US 5,708,146. In another embodiment, (L) comprises a linker residue disclosed in US 6,468,522. In another embodiment, (L) comprises a linker residue disclosed in US 6,103,236. In another embodiment, (L) comprises a linker residue disclosed in US 6,638,509. In another embodiment, (L) comprises a linker residue disclosed in US 6,214,345. In another embodiment, (L) comprises a linker residue disclosed in US 6,759,509. In another embodiment, (L) comprises a linker residue disclosed in WO 2007/103288. In another embodiment, (L) comprises a linker residue disclosed in WO 2008/083312. In another embodiment, (L) comprises a linker residue disclosed in WO 2003/068144. In another embodiment, (L) comprises a linker residue disclosed in WO 2004/016801. In another embodiment, (L) comprises a linker residue disclosed in the document

120 * *. «

WO 2009/134976. In another embodiment, (L) comprises a linker residue disclosed in WO 2009/134952. In another embodiment, (L) comprises a linker residue disclosed in WO 2009/134977. In another embodiment, (L) comprises a linker residue disclosed in WO 2002/08180. In another embodiment, (L) comprises a linker residue disclosed in WO 2004/043493. In another embodiment, (L) comprises a linker residue disclosed in WO 2007/018431. In another embodiment, (L) comprises a linker residue disclosed in WO 2003/026577. In another embodiment, (L) comprises a linker residue disclosed in WO 2005/077090. In another embodiment, (L) comprises a linker residue disclosed in WO 2005/082023. In another embodiment, (L) comprises a linker residue disclosed in WO 2007/011968. In another embodiment, (L) comprises a linker residue disclosed in WO 2007/038658. In another embodiment, (L) comprises a linker residue disclosed in WO 2007/059404. In another embodiment, (L) comprises a linker residue disclosed in WO 2006/110476. In another embodiment, (L) comprises a linker residue disclosed in WO 2005/112919. In another embodiment, (L) comprises a linker residue disclosed in WO 2008/103693. In another mode,

121 (L) comprises a linker residue disclosed in US 6,756,037. In another embodiment, (L) comprises a linker residue disclosed in US 7,087,229. In another embodiment, (L) comprises a linker residue disclosed in US 7,122,189. In another embodiment, (L) comprises a linker residue disclosed in US 7,332,164. In another embodiment, (L) comprises a linker residue disclosed in US 5,556,623. In another embodiment, (L) comprises a linker residue disclosed in US 5,643,573. In another embodiment, (L) comprises a linker residue disclosed in US 5,665,358.

Linkers (L) comprising an auto immolative component can also be used. For example, see US Pat. No. 6,214,345. An example of a self-immolative component is p-aminobenzylcarbamoyl (PABC).

Commercially available linkers can be used in the invention. For example, the commercially available cleavable linker sulfosuccinimidyl 6 [3 '(2-pyridyldithio) -propionamido] hexanoate (sulfo-LC-SPDP: Thermo Pierce Cat # 21650) and the non-cleavable linker succinimidyl 4- [N-maleimidomethyl) ] cyclohexan-1-carboxylate (SMCC: Thermo Pierce Cat # 22360), as demonstrated herein.

122

See also, W02012171020, W02010138719, the range of commercially available linkers, for example, from Concords http://www.concortis.com/home. See also Kim et al., BIOCONJUGATE CHEMISTRY, 21 (8): 1513-1519 August 2010. See also EP2326349. See also copper free click chemical linkers, Angew. Chem. Int. Ed., 2010, 49, p. 9422-9425, ChemBioChem, 2011, 12, p. 1309-1312, http://www.syaaffix.com/techiiology/.

Drug residues (D) (D) is a compound having structure (I), (la) or (Ib) as described herein. It will be recognized by the expert of reasonable ability that the compounds of structure (I), (la) or (Ib) can be appropriately modified to facilitate a conjugation reaction with (L), or if (L) is not present, with ( T), and the formation of a conjugate (T) - (L) - (D) or (T) - (D). Any junction point in (D) can be used. In one embodiment, the C-terminal of (D) forms the junction point in a conjugate (T) - (L) - (D). In another embodiment, the N-terminal of (D) forms the junction point in a conjugate (T) - (L) - (D). In another embodiment, a side chain of (D) forms the junction point in a conjugate (T) (L) - (D).

Novel conjugates comprising peptide toxins

123 microtubule disruptors

In one embodiment of the present disclosure, the conjugates comprising peptide disrupting microtubule toxins covalently linked in the conjugate through the laterial chain of the N-terminal amino acid are provided. In one embodiment, the microtubule disrupting peptide toxin is hemiasterlin or an analog thereof and the toxin covalently binds in the conjugate through the indole residue within the N-terminal amino acid side chain of the toxin peptide. In another embodiment, the microtubule disrupting peptide toxin is HTI-286 or an analog thereof and the toxin covalently binds in the conjugate through the phenyl group within the N-terminal amino acid side chain of the toxin peptide. In one embodiment, the microtubule disrupting peptide toxin is a compound having the structure (I), (la) or (Ib) as described herein.

The compositions object of the present, have anti-mitotic activity and the following structure:

(T) - (L) - (PT) (VII) wherein (T) is a target residue as described herein, (L) is an optional linker as

124 described herein, and (PT) is a microtubule disrupting peptide toxin that covalently binds (L) through the N-terminal amino acid side chain of (PT), or if (L) is not present, (PT) is covalently linked to (T) through the N-terminal amino acid side chain of (PT).

In one embodiment, (T) is an antibody. Accordingly, in one embodiment, antibody-drug conjugates (ADCs) comprising microtubule disrupting peptide toxins that are linked to the conjugate through the N-terminal amino acid side chain are provided.

In one embodiment, (T) - (L) - (PT) has the following structure:

/ N ^R6 ^R2

OR

<img file="MX368258B_D0046.tif" />

where,

Ri and R2 are independently selected from the group consisting of: H and a saturated or unsaturated residue having a linear, branched or non-aromatic cyclic skeleton containing from one to ten carbon atoms, and the carbon atoms are optionally substituted with: -OH, -I, -Br, Cl, -F, -CN, -CO₂H, -CHO, -COSH, or -NO₂;

R3 and R₄ are independently selected from the group that

125 consists of: H, R, ArRo R₃ and R₄ join together to form a ring;

R₃i is selected from the group consisting of: H, R ', ArR-, ArR-Ar, R-Ar-Ar, Ar-Ar-R-, and Ar, where R in each Ar can be substituted, and zero to ten heteroatoms can replace atoms in the chain, for example 0 or S or N can be incorporated into the carbon chain; in one mode, where R '

<img file="MX368258B_D0047.tif" />

m where m is an integer from one to fifteen;

R₆ is selected from the group consisting of: H, R, and ArR-;

R7 and R₈ they are independently selected from the group consisting of: H, R, and ArR-; Y

R₃₂ is selected from:

0 00

II H II IIH II o \ ΛΛ? Ζ — c — N — S — R₁₄ WVY — c — N — S — R₁₄

II IIII

Z —C - ΥκΛΛΛ. O; oO where,

Z is defined as a residue that is selected from the group consisting of: -OH, -0R; -SH; -MR; -NH₂; -NRCH (Rn) COOH; and -NHCH (Rn) COOH, wherein Rn is a residue having the formula: R, or - (CH₂)_nNRi₂Ri3, where n = l-4 and R_i2 and R13 are

126 independently selected from the group consisting of:

H; R; and -C (NH) (NH₂),

Y is defined as a residue selected from the group consisting of: a linear, saturated or unsaturated alkyl group of one to six carbons, optionally substituted with R, ArR-, or X; Y,

X is defined as a residue selected from the group consisting of: -OH, -0R, = 0, = S, -O₂CR, -SH, -SR, -SOCR, -NH₂, NHR, -N (R)₂, -NHCOR, -NRCOR, -I, -Br, -Cl, —F, -CN, -CO₂H, CO₂R, -CHO, -COR, -CONH₂, -CONHR, -CON (R)₂, -COSH, -COSR, NO₂, -SO₃H, -SOR, and —SO₂R;

Ri4 is selected from the group consisting of optionally substituted alkyl, optionally substituted alkylamino, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heterocyclyl and optionally substituted heteroaryls, COR₂₄, CSR_24z -OR₂₄, and -NHR₂₄, where each R₂₄ it is, independently, alkyl optionally substituted with halogen, -OH or -SH;

R is defined as a saturated or unsaturated residue having a linear, branched, or non-aromatic cyclic skeleton containing one to ten carbon atoms, zero to four nitrogen atoms, zero to four oxygen atoms, and zero to four sulfur atoms, and the carbon atoms are

127 optionally substituted with: = 0, = S, OH, -ORio, -O₂CR₁₀, -SH, -SRio, -SOCRio, -NH₂, -NHRio, -N (Ri₀)₂, -NHCORio, -NRi₀CORi₀, I, -Br, -Cl, -F, -CN, -CO₂H, -CO₂Ri₀, -CHO, -Corio, -CONH₂, CONHRio, -CON (R₁₀)₂, -COSH, -COSRiq, -NO₂, -SO₃H, -SOR₁₀, SO₂Riq, where Rio is a linear, branched or cyclic, saturated or unsaturated alkyl group of one to ten carbons;

the ring formed by the union of R₃ and R₄ it is a non-aromatic cyclic skeleton of three to seven members within the definition of R,

Y is defined as a residue selected from the group consisting of: a linear, saturated or unsaturated alkyl group of one to six carbons, optionally substituted with R, ArR-, or X; Y,

X is defined as a residue selected from the group consisting of: -OH, -0R, = 0, = S, -O₂CR, -SH, -SR, -SOCR, -NH₂, NHR, -N (R)₂, -NHCOR, -NRCOR, -I, -Br, -Cl, —F, -CN, -CO₂H, -

<td>CO₂R, -CHO, -COR, -CONH</td><td>₂, -CONHR,</td><td>-CON (R)₂,</td><td>-COSH, -COSR, -</td>

<td>N0₂—SO₃H, -SOR, and</td><td>-SW₂R;</td><td></td><td></td>

<td>or a stereoisomer,</td><td>prodrug</td><td>or salt</td><td>pharmaceutically</td>

<td>acceptable of it.</td><td></td><td></td><td></td>

<td>In one mode, Ar is</td><td>a ring</td><td>aromatic</td><td>selected from</td>

group consisting of: phenyl, naphthyl, anthracil, pyrrolyl.

In one mode, R₃₂ it is:

128

Z — C Υσνν \ where Z and Y are defined as above.

In one mode, R₃₂ it is:

where Z and R_X4 They are defined as above.

In one mode, R₃₂ it is:

R-14 where Y and R_i4 They are defined as above.

In another embodiment, (T) - (L) - (PT) has the following structure:

* 30 GO₁₈ vile¹TL — R ^ Y'N^xY^N'R32 m / N'r, ⁰

Rl7 Rl6 where,

Ris is selected from the group consisting of optionally substituted alkyl, optionally substituted alkylamino, optionally substituted cycloalkyl, optionally substituted aryl, optionally heterocyclyl

129 substituted and optionally substituted heteroaryl;

R16 is selected from the group consisting of H and Ci_ alkyl

6,‘

Ri7 is selected from the group consisting of H and acyl of Ci_ 6 /

Laugh and R30 are independently selected from the group consisting of H, Ci_ alkyl₆ and -SH, with the proviso that

<td colspan="3">both substituents R_i8 and R₃₀ they can not</td><td rowspan="2">be both</td><td rowspan="2">H;</td>

<td>R32 is</td><td colspan="2">select from:</td>

<td></td><td></td><td> 0 0</td><td> 0</td><td> 0</td>

<td></td><td></td><td>II H II</td><td>II</td><td>H II</td>

<td></td><td>0 wvz-</td><td>-c — N — s — R₁₄</td><td>ννχηγ — c—</td><td>-N — S</td>

<td></td><td>II</td><td>II</td><td></td><td>II</td>

<td>z—</td><td>—C - Ywm.</td><td>θ; or</td><td></td><td> 0</td>

where,

Z is defined as a residue that is selected from the group that

<td>consists</td><td>from:</td><td>-OH, -OR; -SH; -</td><td>MR; </td><td>-NH₂; -NRCH (Rn) COOH</td><td>; Y</td>

<td>-NHCH (Rn¡</td><td>iCOOH,</td><td>where Rn is</td><td>a</td><td>residue you have</td><td>the</td>

<td>formula:</td><td>R, 0</td><td>- (CH₂) _nNRi₂Ri3, in</td><td>where</td><td>n = l-4 and Ri₂ and R₁₃</td><td>They are</td>

<td colspan="2">selected</td><td>regardless</td><td>of the</td><td>group consisting</td><td>from:</td>

<td>H; R; Y</td><td></td><td>-C (NH) (NH₂) ,</td><td></td><td></td><td></td>

R is defined as a saturated or unsaturated residue having a linear, branched, or non-aromatic cyclic skeleton containing from one to ten carbon atoms, from zero to four nitrogen atoms, from zero to four oxygen atoms, and from

130 zero to four sulfur atoms, and carbon atoms are optionally substituted with: = 0, = S, OH, -ORi₀, -O₂CR₁₀, -SH, -SRio, -SOCRio, -NH₂, -NHRiq, -N (R_1st)₂, -NHCORiq, -NR₁₀CORi₀, I, -Br, -Cl, -F, -CN, -CO₂H, -CO₂Ri₀, -CHO, -Corio, -CONH₂, CONHRio, -CON (R₁₀)₂, -COSH, -COSRio, -NO₂, -SO₃H, -SORIO, SO₂Riq, where Rio is a linear, branched or cyclic, saturated or unsaturated alkyl group of one to ten carbons;

the ring formed by the union of R₃ and R₄ it is a non-aromatic cyclic skeleton of three to seven members within the definition of R,

Y is defined as a residue selected from the group consisting of: a linear, saturated or unsaturated alkyl group of one to six carbons, optionally substituted with R, ArR-, or X; Y,

X is defined as a residue selected from the group consisting of: -OH, -0R, = 0, = S, -O₂CR, -SH, -SR, -SOCR, -NH₂, NHR, -N (R)₂, -NHCOR, -NRCOR, -I, -Br, -Cl, —F, -CN, -CO₂H, C0₂R, -CHO, -COR, -CONH₂, -CONHR, -CON (R)₂, -COSH, -COSR, N0₂, -SO₃H, -SOR, and —SO₂R;

or a stereoisomer, prodrug or pharmaceutically acceptable salt thereof.

In another embodiment, (T) - (L) - (PT) has the following structure:

131 ^R30 I Ri8

V Λ 1 ¹ τ-L — r ^ Y'n^&gt;Yⁿ'r₃₃ n -N._d^H OR

R17 R₁₆ where,

Ri₅ it is selected from the group consisting of optionally substituted alkyl, optionally substituted alkylamino, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heterocyclyl and optionally substituted heteroaryl;

R16 is selected from the group consisting of H and C6 alkyl /

Ri7 is selected from the group consisting of H and C 1_r 6

Ri8 and R30 are independently selected from the group consisting of H, Ci-g alkyl and -SH, with the proviso that both R substituents₁₈ and R₃₀ they cannot be both H;

R33 is:

where,

Z is as defined above,

R is defined as a saturated or unsaturated residue that has

132 a linear, branched, or non-aromatic cyclic skeleton containing one to ten carbon atoms, zero to four nitrogen atoms, zero to four oxygen atoms, and zero to four sulfur atoms, and carbon atoms are optionally substituted with: = 0, = S, OH, -ORi₀, -O₂CRi₀, -SH, -SR₁₀, -SOCR₁₀, -NH₂, -NHRio, -N (R₁₀)₂, -NHCORiq, -NRi₀COR₁₀, I, -Br, -Cl, -F, -CN, -CO₂H, -CO₂Ri₀, -CHO, -Corio, -CONH₂, CONHRio, -CON (Rio) ₂, -COSH, -COSRio, ~ N0₂, -SO₃H, -SOR₁₀, S0₂Rio, where Rio is a linear, branched or cyclic, saturated or unsaturated alkyl group of one to ten carbons;

the ring formed by the union of R₃ and R₄ it is a non-aromatic cyclic skeleton of three to seven members within the definition of R,

Y is defined as a residue selected from the group consisting of: a linear, saturated or unsaturated alkyl group of one to six carbons, optionally substituted with R, ArR-, or X; Y,

X is defined as a residue selected from the group consisting of: -OH, -0R, = 0, = S, -O₂CR, -SH, -SR, -SOCR, —NH₂, NHR, -N (R)₂, -NHCOR, -NRCOR, -I, -Br, -Cl, —F, -CN, -CO₂H, CO₂R, -CHO, -COR, -CONH₂, -CONHR, -CON (R)₂, -COSH, -COSR, N0₂, -SO₃H, -SOR, and —SO₂R;

or a stereoisomer, prodrug or pharmaceutically salt

133 acceptable of it.

In another embodiment, (T) - (L) - (PT) has the following structure:

<img file="MX368258B_D0048.tif" />

where,

Ri4 is selected from the group consisting of optionally substituted alkyl, optionally substituted alkylamino, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heterocyclyl and optionally substituted heteroaryls, COR₂₄, CSR24, -OR₂₄, _ MR₂₄, and -NHR₂₄, where each R₂₄ it is, independently, alkyl optionally substituted with halogen, -OH or -SH;

R15 is selected from the group consisting of optionally substituted alkyl, optionally substituted alkylamino, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heterocyclyl and optionally substituted heteroaryl;

R16 is selected from the group consisting of H and Ci_ alkyl

6/

134

Ri? is selected from the group consisting of H and C-acyl

Ris and R30 are independently selected from the group consisting of H, Ci_g alkyl and -SH, with the proviso that both R substituents_ig and R₃₀ they cannot be both H;

or a stereoisomer, prodrug or pharmaceutically acceptable salt thereof.

In another embodiment, (T) - (L) - (PT) has the following structure:

<img file="MX368258B_D0049.tif" />

where,

R14 is selected from the group consisting of optionally substituted alkyl, optionally substituted alkylamino, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heterocyclyl and optionally substituted heteroaryls, COR24, CSR24, -OR₂4, - SR24, and -NHR24, where each R₂4 it is, independently, alkyl optionally substituted with halogen, -OH or -SH;

R15 is selected from the group consisting of optionally substituted alkyl, optionally alkylamino

135 substituted, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heterocyclyl and optionally substituted heteroaryl;

Ri₆ is selected from the group consisting of H and Ci_ 6 alkyl /

Ri7 is selected from the group consisting of H and C 1 -C!

Laugh and R30 are independently selected from the group consisting of H, C1-6 alkyl and -SH, provided that both substituents R₁₈ and R30 cannot be both H;

or a stereoisomer, prodrug or pharmaceutically acceptable salt thereof.

In another embodiment, (T) - (L) - (PT) has the following structure:

<img file="MX368258B_D0050.tif" />

where,

R15 is selected from the group consisting of optionally substituted alkyl, optionally substituted alkylamino, optionally substituted cycloalkyl, optionally substituted aryl, optionally heterocyclyl

136 substituted and optionally substituted heteroaryl;

or a stereoisomer, prodrug or pharmaceutically acceptable salt thereof.

In another embodiment, (T) - (L) - (PT) has the following structure:

where,

R is defined as a saturated or unsaturated residue having a linear, branched, or non-aromatic cyclic skeleton containing one to ten carbon atoms, zero to four nitrogen atoms, zero to four oxygen atoms, and zero at four sulfur atoms, and the carbon atoms are optionally substituted with: = 0, = S, OH, -ORi₀, -O₂CRi₀, -SH, -SRio, -SOCRio, -NH₂, -NHR₁₀, ~ N (R_1st)₂, -NHCOR₁₀, -NR₁₀CORi₀, I, -Br, -Cl, -F, -CN, -CO₂H, -CO₂Ri₀, -CHO, -COR_W, -CONH₂, CONHRio, -CON (Rio) ₂, -COSH, -COSRio, -N0₂, -SO₃H, -SOR₁₀, SO₂Riq, where Rio is a linear, branched or cyclic, saturated or unsaturated alkyl group of one to ten carbons;

or a stereoisomer, prodrug or pharmaceutically acceptable salt thereof.

In another mode, (T) - (L) - (PT) has the following

137 structure:

<img file="MX368258B_D0051.tif" />

where,

R is defined as a saturated or unsaturated residue having a linear, branched, or non-aromatic cyclic skeleton containing from one to ten carbon atoms, from zero to four nitrogen atoms, from zero to four oxygen atoms, and from zero at four sulfur atoms, and the carbon atoms are optionally substituted with: = 0, = S, OH, -OR₁₀, -O₂CRiq, -SH, -SR₁₀, -SOCRIO, -NH₂, -NHR₁₀, -N (R₁₀)₂, -NHCORio, -NRi₀CORi₀, I, -Br, -Cl, -F, -CN, -CO₂H, -CO₂R₁₀, -CHO, -CORi₀, -CONH₂, CONHRio, -CON (Rio) ₂, -COSH, -COSR₁₀, -N0₂, -SO₃H, -SOR₁₀, SO₂R₁₀, where Rio is a linear, branched or cyclic, saturated or unsaturated alkyl group of one to ten carbons;

or a stereoisomer, prodrug or pharmaceutically acceptable salt thereof.

In a further embodiment of the invention, (PT) is a hemisterlin analog, such as those described in US 7,579,323, which is incorporated herein by reference in its entirety for all purposes.

In the synthesis of conjugates, including ADCs, which comprises

138 the microtubule disrupting peptide toxins, the peptide bond through the Nterminal amino acid side chain has several advantages. As demonstrated herein, the side chains of such peptide toxins are sensitive to chemical modifications and manipulations that facilitate the formation of covalently bound conjugates without compromising potency. As demonstrated herein, such conjugates are potent cytotoxic compositions capable of delivering payloads of peptide toxin.

Administration

For administration purposes, the compounds of the present specification may be administered as a crude chemical or may be formulated as pharmaceutical compositions. The pharmaceutical compositions of the present specification comprise a compound of structure (I), (la) or (Ib) and a pharmaceutically acceptable carrier, diluent or excipient. The compound of structure (I), (la) or (Ib) is present in the composition in an amount that is effective to treat a particular disease or condition of interest - for example, in an amount sufficient to treat cancer or growth. of tumor cells, and preferably with acceptable toxicity

139 for the patient The activity of the compounds of structure (I), (la) or (Ib) can be determined by one skilled in the art, for example, as described in the Examples below. Appropriate concentrations and dosages can be readily determined by one skilled in the art.

The administration of the compounds of the description, or their pharmaceutically acceptable salts, in pure form or in an appropriate pharmaceutical composition, can be carried out through any of the accepted modes of administration of agents that serve similar utilities. The pharmaceutical compositions of the specification can be prepared by combining a compound of the specification with a pharmaceutically acceptable carrier, diluent or excipient, and can be formulated in preparations in solid, semi-solid, liquid or gaseous forms, such as tablets, capsules , powders, granules, ointments, solutions, suppositories, injections, inhalants, gels, microspheres, and aerosols. Typical routes of administration of such pharmaceutical compositions include, without limitation, oral, topical, transdermal, inhalation, parenteral, sublingual, buccal, rectal, vaginal, and intranasal. The term parenteral, as used in this

140 Document includes subcutaneous, intravenous, intramuscular injections, intrasternal injection or infusion techniques. The pharmaceutical compositions of the specification are formulated to allow the active ingredients contained therein to be bioavailable upon administration of the composition to a patient. The compositions to be administered to a subject or patient take the form of one or more dosage units, where for example, a tablet may be a single dosage unit, and a container of a compound of the description in the form of an aerosol may contain a plurality of dosage units. Actual methods for preparing such dosage forms are known, or will be apparent, to those skilled in this art; for example, see Remington: The Science and Practice of Pharmacy, 20th edition (Philadelphia College of Pharmacy and Science, 2000). The composition to be administered, in any case, contains a therapeutically effective amount of a compound of the specification, or a pharmaceutically acceptable salt thereof, for the treatment of a disease or condition of interest according to the teachings of this specification. A pharmaceutical composition of the specification may be in the form of a solid or liquid. In one aspect, the vehicles

141 they are particulate, so that the compositions are, for example, in tablet or in powder form. The vehicle (s) may be liquid, the compositions being, for example, an oral syrup, injectable liquid or an aerosol, which is useful in, for example, administration by inhalation.

When intended for oral administration, the pharmaceutical compositions of the present specification are typically in either solid or liquid form, where semi-solid, semi-liquid, suspension and gel forms are included within the forms considered herein. as solids or liquids.

As a solid composition for oral administration, the pharmaceutical compositions can be formulated in a powder, granule, compressed tablet, pill, capsule, chewing gum, wafer or the like. Said solid composition will typically contain one or more inert diluents or edible carriers. In addition, one or more of the following may be present: binders such as carboxymethyl cellulose, ethyl cellulose, microcrystalline cellulose, gum tragacanth or gelatin; excipients such as starch, lactose or dextrins, disintegrating agents such as alginic acid, sodium alginate, Primogel, corn starch and the like; lubricants such as

142 magnesium stearate or Sterotex; sliders such as colloidal silicon dioxide; sweetening agents such as sucrose or saccharin; a flavoring agent such as peppermint, methyl salicylate or orange flavor; and a coloring agent.

When the pharmaceutical composition is in the form of a capsule, for example, a gelatin capsule, it may contain, in addition to the materials of the above type, a liquid carrier such as polyethylene glycol or oil.

The pharmaceutical compositions of the description may be in the form of a liquid, for example, an elixir, syrup, solution, emulsion or suspension. The liquid may be for oral administration or for injection delivery, as two examples. When intended for oral administration, the pharmaceutical compositions of the description typically contain, in addition to the present compounds, one or more of a sweetening agent, preservatives, dye / dye and flavor enhancer. In a composition intended to be administered by injection, one or more of a surfactant, preservative, wetting agent, dispersing agent, suspending agent, buffer, stabilizer and isotonic agent may be included.

The liquid pharmaceutical compositions of the description, whether solutions, suspensions or other similar forms,

143 they may include one or more of the following adjuvants: sterile diluents such as water for injection, saline solution, preferably physiological saline solution, Ringer's solution, isotonic sodium chloride, fixed oils such as synthetic mono or diglycerides that can serve as the solvent medium or suspension, polyethylene glycols, glycerin, propylene glycol or other solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose. Parenteral preparations can be enclosed in ampoules, disposable syringes or multi-dose vials made of glass or plastic. The physiological saline solution is a preferred adjuvant. An injectable pharmaceutical composition is preferably sterile.

A liquid pharmaceutical composition of the specification intended for administration either orally or parenterally must contain an amount of a compound of the specification such that an adequate dosage will be obtained.

144

The pharmaceutical compositions of the specification may be intended for topical administration, in which case the vehicle may suitably comprise a solution, emulsion, ointment or gel base. The base, for example, may comprise one or more of the following: petrolatum, lanolin, polyethylene glycols, beeswax, mineral oil, diluents such as water and alcohol, and emulsifiers and stabilizers. Thickening agents may be present in a pharmaceutical composition for topical administration. If intended for transdermal administration, the composition may include a transdermal patch or iontophoresis device.

The pharmaceutical compositions of the specification may be intended for rectal administration, in the form, for example, of a suppository, which will melt in the rectum and release the drug. Compositions for rectal administration may contain an oil base as a suitable non-irritating excipient. Such bases include, without limitation, lanolin, cocoa butter and polyethylene glycol.

The pharmaceutical compositions of the specification may include various materials, which modify the physical form of a solid or liquid dosage unit. For example, the composition may include materials that form a shell of

145 coating around the active ingredients. The materials that form the shell are typically inert, and can be selected from, for example, sugar, shellac, and other enteric coating agents. Alternatively, the active ingredients can be encapsulated in a gelatin capsule.

The pharmaceutical compositions of the specification can be prepared in dosage units that can be administered as an aerosol. The term aerosol is used to indicate a variety of systems ranging from those of colloidal nature to systems consisting of pressurized packages. The supply can be by a liquefied or compressed gas or by a suitable pumping system that dispenses the active ingredients. The aerosols of compounds of the description can be supplied in single-phase, two-phase or three-phase systems in order to supply the active ingredient (s). The aerosol supply includes the necessary container, activators, valves, subcontainers, and the like, which together can form a kit. A person skilled in the art, without having done experimentation, can determine preferred aerosols.

The pharmaceutical compositions of the specification can be prepared by methodology well known in the

146 Pharmaceutical technique For example, a pharmaceutical composition intended to be administered by injection can be prepared by combining a compound of the specification with sterile distilled water in order to form a solution. A surface active agent can be added to facilitate the formation of a homogeneous solution or suspension. Surfactants are compounds that interact non-covalently with the compound of the specification in order to facilitate the homogeneous dissolution or suspension of the compound in the aqueous delivery system.

The compounds of the specification, or their pharmaceutically acceptable salts, are administered in a therapeutically effective amount, which will vary depending on a variety of factors including the activity of the specific compound employed; the metabolic stability and duration of action of the compound; the age, body weight, general health, sex and diet of the patient; the mode and time of administration; the excretion rate; the combination of drugs; the severity of the particular disorder or condition; and the subject undergoing therapy.

The compounds of the specification, or pharmaceutically acceptable derivatives thereof, can also be administered simultaneously with, before, or after the

147 administration of one or more other therapeutic agents. Such combination therapy includes the administration of a single pharmaceutical dosage formulation containing a compound of the specification and one or more additional active agents, as well as the administration of the compound of the specification and each active agent in its own separate pharmaceutical dosage formulation. . For example, a compound of the specification and the other active agent can be co-administered to the patient in a single oral dosage composition such as a tablet or capsule, or administered each agent in separate oral dosage formulations. When separate dosage formulations are used, the compounds of the specification and one or more additional active agents can be administered essentially at the same time, that is, concurrently, or in separate stepped times, that is, sequentially; Combination therapy is understood to include all these regimens.

It is understood that in the present description, combinations of substituents and / or variables of the formulas represented are permissible only if such contributions result in stable compounds.

148

It will also be appreciated by those skilled in the art that in the synthesis processes described herein the functional groups of intermediate compounds may need to be protected by suitable protecting groups. Such functional groups include hydroxy, amino, mercapto and carboxylic acid. As described above, suitable hydroxy protecting groups include trialkylsilyl or diarylalkyl silyl (for example, t-butyldimethylsilyl, tbutyldiphenylsilyl or trimethylsilyl), tetrahydropyranyl, benzyl, and the like, and suitable protecting groups for amino, amidino and guanidino include t-butoxycarbonyl benzyloxycarbonyl, and the like. Suitable protecting groups for mercapto include -C (O) -R (where R is alkyl, aryl or arylalkyl), p-methoxybenzyl, trityl and the like. Suitable protecting groups for carboxylic acid include alkyl, aryl or arylalkyl esters. The protecting groups can be added or removed according to standard techniques, which are known to one skilled in the art and as described herein. The use of protective groups is described in detail in Green, TW and PGM Wutz, Protective Groups in Organic Synthesis (1999), 3^to Ed. Wiley. As one skilled in the art will appreciate, the protecting group may also be a polymer resin such as a resin.

149

Wang, Rink resin or a 2-chlorotrityl chloride resin.

It will also be appreciated by those skilled in the art, although a protected derivative of compounds of this description may not possess pharmacological activity as such, they can be administered to a mammal and subsequently metabolized in the body to form compounds of the description, which are pharmacologically active. Therefore, such derivatives can be described as prodrugs. All prodrugs of the compounds of this description are included within the scope of the present description.

In addition, the compounds of the description that exist in free base or acid form can be converted into their pharmaceutically acceptable salts by treatment with the appropriate inorganic or organic base or acid by methods known to one skilled in the art. The salts of the compounds of the description can be converted to their free base or acid form by standard techniques.

The following examples illustrate various methods of manufacturing compounds of this description, that is, compounds of structures (I), (la), (Ib), (VI) and (VII). It is understood that one skilled in the art may be able to make these compounds by similar methods or by combining other known methods for a

150 skilled in the art. It is also understood that one skilled in the art would be able to make, in a similar manner as described below, other compounds of structure (I), (la), (Ib), (VI) or (VII) not specifically illustrated then , by using the appropriate starting components and modifying the synthesis parameters, as necessary. In general, the starting components can be obtained from sources such as Sigma Aldrich, Lancaster Synthesis, Inc., Maybridge, Matrix Scientific, TCI, and Fluorochem USA., Etc. or synthesized according to sources known to those skilled in the art (see, for example, Advanced Organic Chemistry: Reactions, Mechanisms and Structure, 5th edition (Wiley, December 2000)), or prepared as described herein.

The following examples are provided for purposes of illustration, not limitation.

EXAMPLES

GENERAL SYNTHETIC SCHEMES

GENERAL SCHEME

151

<td>X z- 0 ^S</td><td>fi 0 D 'Ά'</td>

<td>rvV'n</td><td>Procedure and^z LJ vg ⁰ gP «- ___________X _to í '' ^ &lt;Y 'Ν' V '' V '' N '* R ° Senerail ^{&gt; R}^ U_X " ,TO". I</td>

<img file="MX368258B_D0052.tif" />

<img file="MX368258B_D0053.tif" />

r /^NBoc \, .CO₂Et

Procedure |

-----------&gt; ^R·

Segallo ^RZ

..COJEt

Rn

Process

Gender! eleven "_zNBoc R¿

BOCHN

COJEt

<img file="MX368258B_D0054.tif" />

RX

Process

General 12 R 'SO;? NH_;

<img file="MX368258B_D0055.tif" />

A £ b. CH_;Cl; _β

<img file="MX368258B_D0056.tif" />

ΤΓ

<img file="MX368258B_D0057.tif" />

Ό—

To OH

Ar

<img file="MX368258B_D0058.tif" />

OH _BlN'-N

or. or

General Procedure ^ 'OH \ Z 1 Pd ^ dbah

- ................------- Procedure ESaf / d ^ f 'S0₂nh₂ r^wm'^ÜM

<img file="MX368258B_D0059.tif" />

BoeHN

one. PÓ.H ?, AcOH

two. BOQO RfeN

<img file="MX368258B_D0060.tif" />

OM «

Process

General 3

BncHN γ oh, N8oc θΡ cr^H.HNco

one. t-BuOH

two. DMAP

N 'ÑBoc

one. NHRiRo

two. TFA

OO ^Ri'N'^S'NH₂ r₂

General Procedure 14

<img file="MX368258B_D0061.tif" />

152

General Procedure 1 - Trifluoroacetamide Installation

Ά a stirred suspension of the amine in 1,4-dioxane was added trifluoroacetic anhydride (1.1 equivalents). The reaction mixture passed from a suspension to a solution and back to a suspension again. The progress of the reaction was monitored by TLC and / or HPLC-MS until its conclusion. Once the starting material was completely consumed, the reaction was diluted with hexanes or diethyl ether, filtered on a Buchner funnel and the resulting solids dried under reduced pressure to give pure trifluoroacetamide.

General procedure 2 - DCC / DMñP-mediated N-acylsulfonamide formation

To a stirred solution of the acid in dichloromethane was added a solution of the sulfonamide (1.3 equivalents, in dichloromethane, N, N-dimethylformamide, or a mixture thereof, as necessary). Dicyclohexylcarbodiimide (1.2 equivalents) and then N, N-dimethylaminopyridine (1.2 equivalents) were added. The course of the reaction was monitored by HPLC-MS (typically 16 h) and excess by-products could be precipitated by the addition of diethyl ether. The solids were filtered off and washed with 1: 1 diethyl ether / dichloromethane. The combined organic layers are

153 concentrated, and the residue was purified by silica gel chromatography or optionally HPLC-preparative to give the desired Nacylsulfonamide.

General procedure 3 - general saponification

To a solution of the trifluoroacetamide or ester containing the construction in 1,4-dioxane or methanol was added lithium hydroxide (10 equivalents) and water (10% v / v). The reaction was allowed to stir at room temperature or optionally heated to 50 ° C. The course of the reaction was monitored by HPLC-MS. At the end, the volatiles were removed under reduced pressure, the pH was adjusted from the aqueous layer if necessary and washed successively with dichloromethane or acetate

<td>of ethyl.</td><td>Organic phases</td><td>they combined,</td><td>I know</td><td colspan="2">they dried over</td>

<td>MgSO₄, I know</td><td>leaked and it</td><td>concentrated.</td><td>The</td><td>product</td><td>from</td>

<td>reaction</td><td>it was used either</td><td>how that</td><td>I know</td><td>purified</td><td>by</td>

silica gel chromatography as necessary.

General procedure 4 - HATU-mediated peptide bond formation

To a stirred solution of the carboxylic acid in a minimum amount of dichloromethane or N, N-dimethylformamide or a mixture thereof, at 0^or C HATU (equivalents) and N, Ndiisopropylethylamine (4 equivalents) were added. Stirring was continued for a brief induction period (5-20 minutes) in which

154 At that time the reaction was charged with a solution of the amine in dichloromethane. The reaction was allowed to warm to room temperature and the progress was monitored by HPLC-MS. At the end, the volatiles were removed under reduced pressure and the residual material was purified by silica gel chromatography or reverse phase HPLC to provide the amide in the appropriate purity.

General procedure 7 - elimination of the Boc group

To a solution of the Boc-protected construction in dichloromethane was added 10% v / v trifluoroacetic acid. The course of the reaction was monitored by HPLC-MS. At the end, all volatiles were removed under reduced pressure. The residual material was purified either by reverse phase HPLC, silica gel chromatography or precipitation from a mixture of methanol / dichloromethane / cold diethyl ether.

General Procedure 8 - Suzuki Catalyzed Cross Coupling with Pd

A suspension of aryl bromide, aryl (or alkenyl) boronic acid (1.5 eq), Pd (OAc)₂ (10% in mol), 2- (di-tertbutylphosphine) biphenyl (20% in mol), and K3PO₄ (3 eq) in THE stirred under N₂ at room temperature for 16h (or 50 ° C for 2 h). The resulting brown reaction mixture is

155 * '' I ♦ ís. w 1 *&gt; »diluted with ether and washed with 1M NaOH (3x). The aqueous washings were combined and extracted with ether (2x). The organic extracts were combined, dried over MgSO₄, filtered, concentrated in vacuo and purified by silica gel column chromatography (eluting with MeOH / CH mixtures₂C1₂) to provide the cross coupling product.

General procedure 9 - Cross coupling of magnet magnetized with Cu (methoxy installation)

A mixture of aryl bromide, CuBr (20% in mol), NaOMe (20 eq, 4.9 M in MeOH) and EtOAc (1.5 eq) was stirred under N₂ at 95 ° C for 16h. The resulting mixture was diluted with H₂0 and poured into cold 1M citric acid (0^or C) in agitation. After stirring for 10 min, the mixture was extracted with EtOAc (4x). The organic extracts were combined, washed with H₂O (2x) and brine (lx), dried over MgSO₄, filtered and concentrated in vacuo. The product was used in the next step without further purification.

General Procedure 10 - synthesis of vinyl ester amino esters

The procedure for the synthesis of Weinreb amide, reduction and subsequent olefination thereof was used as described by Nieman JA et al. J. Nat. Prod. 2003, 66, 183-199 for commercially desired amino acids without

156 modifications.

General Procedure 11 - Establishment of amino acid vinylgo Boc-t-Leucin- (Me)

The amino vinyl ester was deprotected and coupled to Boc-tleucine according to procedures described by Nieman JA et al. J. Nat. Prod. 2003, 66, 183-199 without modifications.

General Procedure 12 - formation of snl fnnami from alkyl halide

To a suspension of the desired alkyl halide in 2: 1 of H₂O / EtOH sodium sulfite (1.2 equiv) was added. The resulting mixture was heated at reflux for 6-24h. Then, the reaction was cooled to room temperature, the solvents were removed under reduced pressure to remove ethanol and the product precipitated. The sodium alkylsulfonate was filtered, pooled and dried in vacuo. These solids were then suspended in dichloromethane and phosphorus pentachloride (2 equiv) was added with stirring. The resulting suspension was heated at reflux for 2h and allowed to cool to room temperature. The reactions were cooled to 0^or C and be added water drop by drop to consume excess phosphorus pentachloride. The mixture was transferred to a separatory funnel and the organic phase was washed with brine, dried over MgSOi, filtered and concentrated to give the desired sulfonyl chloride. The

157 derivative chloride thus was subsequently dissolved in THF and added dropwise to a stirred aqueous solution of concentrated ammonium hydroxide at 0^or C. Upon completion of the addition, the reaction was concentrated under reduced pressure and diluted with water and ethyl acetate. The organic phase was washed with brine, dried over MgSO₄, was filtered and concentrated to give the desired sulfonamide in the purity sufficient for later use.

General Procedure 13 - formation of sulfonamide from substituted aryl compounds

Ά A stirred mixture of the desired substituted aryl compound in chloroform was added chlorosulfonic acid (4 equiv). The reaction was heated at 70 ° C for 1 h and allowed to cool to room temperature. Thionyl chloride (2 equiv) was added and the reaction was heated again at 70 ° C for 1 h. The content of the reaction vessel was concentrated under reduced pressure to give an oil that was subsequently dissolved in duplicate in toluene and concentrated under reduced pressure to remove residual acid. The remaining material was dissolved in THF and added dropwise to a concentrated, stirred solution of ammonium hydroxide at 0^or C. Upon completion of the addition, the reaction was concentrated under reduced pressure and the residue was partitioned between acetate.

158 ethyl and water. The organic phase was washed with brine, dried over MgSO₄, filtered and concentrated to give the desired phenylsulfonamide in purity suitable for later use.

General Procedure 14 - formation of sn 7 -Famam-i fia

The procedures used to generate the desired sulfamamides were adapted from Winum, J.-Y. et al., Org Lett, 2001, 3 (14), 2241-2243.

General Procedure 15 - Preparation of MC-VC-PABC toxins

The appropriate intermediate amine or aniline was collected in DMF (~ 90 mg / mL), to which 1-hydroxybenzotriazole hydrate (0.3 eq) was added, then MCVC-PABC-PNP (4 - ((R) -2- ( (R) -2- (6- (2,5-dioxo-2,5-dihydro-lHpyrrol-l-yl) hexanamido) -3-methylbutanamido) -5ureidopentanamido) benzyl 4-nitrophenyl carbonate) obtained commercially (1.3 eq) as described in Firestone, et al. US6214345 followed by pyridine (25 eq). The reaction was covered to protect it from light and stirred at room temperature for 24 to 48h. The reaction mixture could be purified by concentration of the mixture and performing flash chromatography directly on the crude, or, alternatively, it could be diluted with DMSO to an appropriate volume and injected directly into a preparative HPLC

159 to give the pure MC-VC-PABC-R construction.

All sulfonamides and sulfanamides or precursor compounds to the materials used in the following procedures were purchased commercially and handled, if necessary, in such a way that they are suitable for use. Specifically, general procedures 1, 12, 13 and 14 were used to handle commercially available starting materials unless otherwise indicated below. Sulfamamide analogs of Nacylsulfonamide containing compounds described herein can be synthesized by the technician with reasonable knowledge in the art based on the teachings of this document and knowledge in the art, and are included within the scope of the invention.

REPRESENTATIVE COMPOUNDS

Example 1

Br / nh₂

-bromopropan-l-sulfonamide

To a stirred suspension of potassium bromide (1904g) in water (2.8 ml) was added 1,3-propanesulfone. The reaction was heated to

60 ° C with stirring for 1 hour and allowed to cool to room temperature. Ethanoi (~ 45 mi) was added

160 with stirring and a precipitate formed. The suspension was filtered on a Buchner funnel and the solids were collected and dried under high vacuum overnight to give potassium 3-bromopropan-lsulfonate (2.90 g, 12.0 mmol) as a white solid.

The above solid was added to a round bottom flask equipped with a stir bar. Phosphorus pentachloride (3.22 g, 1.3 equiv) was added in a single charge and the flask was gently shaken to mix the solids. It was observed that a gas formed and the solids became slightly molten. A single drop of water was added to the mixture and a vigorous evolution of gas was observed, with a significant melting of the reaction mixture. The flask was immersed in an oil bath at 70 ° C and the molten mixture was manipulated to try to make it as uniform as possible. After 10 minutes of heating, the flask was allowed to cool to room temperature and charged with ice (~ 60 ml) and diethyl ether (~ 80 ml) and stirred vigorously. The biphasic mixture was transferred to a separatory funnel, the organic layer was washed with brine, then dried over MgSO₄, filtered and concentrated to a total volume of ~ 25 ml. The ether layer was added to a 100 ml round bottom flask, a stir bar was added and the flask was cooled to 0 ° C

161 In an ice bath. Ammonium (NH₄OH, 28% bc, 5 ml) with vigorous stirring and an emulsion formed. After the emulsion had calmed down, brine (~ 20 mL) and diethyl ether (~ 20 mL) were added and the mixture was transferred to a separatory funnel. The organic phase was separated, dried over MgSO₄ and concentrated to give the title compound as a rigid syrup which solidified at rest (0.782g).

^:H NMR (400MHz, DMSO-d6) δ (ppm) = 2.24 (p, 2H, J = 6.5 Hz), 3.12 (t, 2H, J = 6.5 Hz), 3.66 (t, 2H, J = 6.5 Hz), 6.91 (s, 2H).

Example 2

<img file="MX368258B_D0062.tif" />

3- (tritylthio) propan-l-sulfonamide

To a stirred solution of triphenylmethantiol (0.276g) in N, N-dimethylformamide at 0^or C, sodium hydride (0.04g, 1 equiv) was added. After the effervescence ceased, 3bromopropan-l-sulfonamide (O.lOOg, 0.5 equiv) was added as a solid in a single portion and the reaction was allowed to warm to room temperature. The progress of the reaction was monitored by HPLC-MS and TLC (40% EtOAc in hexanes). After 2h, the

162 The reaction was quenched with water (~ 0.5 mL) and concentrated on a rotary evaporator under high vacuum. The resulting oil was partitioned between ethyl acetate and brine, transferred to a separatory funnel and the organic phase was washed with brine, dried over MgSO.₄, was concentrated and purified by flash chromatography (5-50% EtOAc in hexanes) to give the title compound (0.135 g) as a white crystalline solid.

NMR (400MHz, CD3OD) δ (ppm) = 1.77-1.85 (m, 2H), 2.35 (t, 2H, J = 6.5 Hz), 2.95-2.99 (t, 2H, J = 6.5 Hz), 7.22-7.33 ( m, 9H), 7.40-7.45 (m, 6H).

Example 3

Acid (6S, 9S, 12S, E) -9-tert-butyl-12-isopropyl-2,2,5,11,14pentamethyl-4,7,10-trioxo-6- (2-phenylpropan-2-yl) -3-oxa-5, 8, 11 triazapentadec- 13-in-15-oic

Synthesized according to Nieman JA et al. J. Nat. Prod. 2003, 66, 183-199.

Example 4

SH

163 (S, E) -N- (3-mercaptopropylsulfonyl) -2,5-dimethyl-4 - ((S) -N, 3, 3-trimethyl-2 - ((S) -3-methyl-2- (methylamino) - 3-phenylbutanamide) butanamide) hex-2-enamide (Compound A)

Example 4 was synthesized from Examples 2 and 3 in accordance with General Procedures 2 and 7 with the inclusion of tri-isoproipsilane (2 eq) with Procedure 9.

NMR (400MHz, CD3OD) δ (ppm) = 0.88 (3H, d, J = 6.2 Hz),

0.94 (3H, d, J = 6.2 Hz), 1.08 (s, 9H), 1.40 (s, 3H), 1.48 (s, 3H), 1.94 (d, 3H, J = 1.29 Hz), 2.03-2.16 (m , 3H), 2.41 (s, 3H), 2.67 (t, 2H, J = 9.76 Hz), 3.16 (s, 3H), 3.46-3.50 (m, 2H), 4.08 (br s, 1H), 4.94 (s , 1H), 5.07 (t, 1H, J = 10.0 Hz), 6.59 (d, 1H, J = 9.5 Hz), 7.32-7.37 (m, 1H), 7.41-7.48 (m, 2H), 7.50-7.57 ( m, 2H).

The methods described above were used to generate the following analogous compounds.

Example 5

or. _ZOR

2,2'-disulfanediildietansulfonamide

Synthesized according to Lemaire, H. and Rieger, M in J.

Org. Chem. 1961, 1330-1331.

164

Example 6

<img file="MX368258B_D0063.tif" />

(S, E) -N- (2-mercaptoethylsulfonyl) -2,5-dimethyl-4 - ((S) -N, 3,3-trimethyl-2 - ((S) -3-methyl-2- (methylamino) - 3-phenylbutanamide) butanamide) hex-2-enamide (Compound B)

To a solution of acid (6S, 9S, 12S, E) -9-tert-butyl-12isopropyl-2,2,5, 11,14-pentamethyl-4,7,10-trioxo-6- (2phenylpropan-2- il) -3-oxa-5,8, ll-triazapentadec-13-en-15-oic (0.138g, 2.4 equiv) in dichloromethane (4ml)) was added 2,2'disulfanediildietansulfonamide (0.028g), diisopropylcarbodiimide (0.044 ml, 2.4 equiv) and N, Ndimethylpyridine (0.034g, 2.8 equiv). Stirring was continued for 16 h until the time when TLC analysis (5% MeOH (with 5% AcOH) in 70/30 Cl ^ Cla / hexanes) indicated the complete consumption of disulfanedisulfonamide. The reaction was diluted with hexanes (~ 5 mL), filtered to remove solids, concentrated and the resulting oil was purified by flash chromatography.

Next, the chromatographically purified materials were dissolved in dichloromethane (3 ml), a stir bar was added, then acid

165 trifluoroacetic (0.60 ml) and tri-isopropylsilane (0.20 ml). The mixture immediately turned yellow, fading over 5 minutes and conversion of the material into the desired product, which was monitored by HPLC-MS. After complete conversion, the reaction was concentrated to dryness and the residue was purified by flash chromatography (0-15% MeOH (containing 5% AcOH) in 80/20 CH2C12 / hexanes). HPLC-MS showed that this isolate is a mixture of free thiol and disulfide.

^ςΗ NMR (400MHz, CD3OD) δ (ppm) = 0.88 (3H, d, J = 6.2 Hz), 0.93 (3H, d, J = 6.2 Hz), 1.07 (s, 9H), 1.40 (s, 3H), 1.47 (s, 3H), 1.91-2.05 (m, 5H), 2.32 (s, 3H), 2.67 (t, 2H, J = 9.76 Hz), 3.07-3.18 (m, 5H), 3.52-3.59 (m, 2H), 3.85 (s, 1H), HH 4.08 (br s, 1H), 4.93 (s, 1H), 5.09 (t, 1H, J = 10.0 Hz), 6.76 (d, 1H, J = 9.5 Hz), 7.29-7.35 (m, 1H), 7.39-7.46 (m, 2H), 7.49-7.5S (m, 2H). C29H48N4O5S2 caled. [M + H]⁺ = 598.15 amu; found m / z = 598.16.

Example 7

O s-nh₂

4- (tritylthiomethyl) benzenesulfonamide

To a stirred solution of triphenylmethantiol (0.276g, 2 eq) in

N, N-dimethylformamide (3 ml) at 0^or C sodium hydride was added

166 (60% w / w dispersion in mineral oil, 0.04g, 2 equiv). When the effervescence had ceased, 4 (bromomethyl) benzenesulfonamide (0.125g, 1 equiv) was added in a single portion and the reaction was allowed to warm to room temperature. HPLC-MS at 20 minutes indicated that the conversion was complete. The reaction was quenched with acetic acid (~ 0.2 ml), concentrated to dryness in vacuo and the subsequent residue was partitioned between ethyl acetate and brine. The organic layer was separated, dried over MgSO₄, was filtered, concentrated and purified by flash chromatography (050% ethyl acetate in hexanes). Fractions containing the desired material were concentrated to dryness to provide the desired compound as a colorless solid (0.200 g).

⁴Η NMR (400MHz, DMSO-d6) δ (ppm) = 3.38 (s, 2H), 7.24-7.35 (m, 7H), 7.36-7.44 (m, 12H), 7.67-7.73 (m, 2H)

Example 8

<img file="MX368258B_D0064.tif" />

(S, E) -N- (4- (mercaptomethyl) phenylsulfonyl) -2,5-dimethyl-4- ((S) N, 3,3-trimethyl-2 - ((S) -3-methyl-2- (methylamino) -3-phenylbutanamide) butanamide) hex-2-enamide (Compound C).

167

The title compound is prepared in accordance with Examples 3 and 7 according to General Procedures 2 and 7 ^XH NMR (400MHz, CD3OD) δ (ppm) = 0.88 (d, 3H, J = 6.2 Hz),

0.91 (d, 3H, J = 6.2 Hz), 1.06 (s, 9H), 1.38 (s, 3H), 1.47 (s, 3H), 1.86 (s, 3H), 1.99-2.05 (m, 1H), 2.41 (s, 3H), 2.67 (t, 2H, J = 9.76 Hz), 3.14 (s, 3H), 3.80 (s, 2H), HH 4.10 (br s, 1H), 4.93 (s, 1H), 5.00 ( t, 1H, J = 10.0 Hz), 6.54 (d, 1H, J = 9.5 Hz), 7.30-7.51 (m, 5H), 7.52-7.58 (m, 2H), 7.90-7.97 (m, 2H). C34H50N4O5S2 caled. [M + H]⁺ = 659.25 amu; found m / z = 659.37.

Example 9 (S, E) -2,5-dimethyl-N-tosyl-4 - ((S) -N, 3,3-trimethyl-2 - ((S) -3-methyl-2- (methylamino) - 3-phenylbutanamide) butanamide) hex-2enamide {Compound D)

The title compound is prepared according to Example 3 and tosylsulfonamide using General Procedures 2 and ^XH NMR (400MHz, CD3OD) δ (ppm) = 0.88-0.94 (m, 6H), 1.06 (s,

9H), 1.35 (s, 3H), 1.45 (s, 3H), 1.86 (s, 3H), 2.02-2.11 (m,

1H), 2.44 (s, 3H), 2.51 (s, 3H), 3.17 (s, 3H), HH 4.35 (s,

168

1Η), 4.89-4.99 (m, 2H), 6.48 (d, 1H, J = 9.5 Hz), 7.30-7.43 (m, 4H), 7.43-7.50 (m, 2H), 7.51-7.57 (m, 2H) . Ο₃₄Η₅₀Ν₄Ο₅3 caled [M + H]⁺ = 627.15 amu; found m / z = 627.31.

Example 10

<img file="MX368258B_D0065.tif" />

(S, E) -2,5-dimethyl-N- (methylsulfonyl) -4 - ((S) -N, 3,3-trimethyl-2 ((S) -3-methyl-2- (methylamino) -3 -phenylbutanamide) butanamide) hex2-enamide {Compound E)

The title compound is prepared according to Example 3 and methanesulfonamide using General Procedures 2 and ^ΤΗ NMR (400MHz, CD3OD) δ (ppm) = 0.87-0.98 (3H (m, 6H), 1.09 (s, 9H), 1.40 (s, 3H), 1.49 (s, 3H), 1.97 (s, 3H) , 2.03-2.13 (m, 1H), 2.52 (s, 3H), 2.67 (t, 2H, J = 9.76 Hz), 3.18 (s,

3H), 3.31 (s, 3H), 4.38 (s, 1H), 4.94 (d, 1H, J = 8.2 Hz),

5.07 (t, 1H, J = 10.0 Hz), 6.54 (d, 1H, J = 9.5 Hz), 7.307.40 (m, 1H), 7.40-7.51 (m, 2H), 7.51-7.59 (m, 2H) .

C₂₈H₄₆N₄OR₅S caled. [M + H]⁺ = 551.30 amu; found m / z =

551.34.

169

Example 11

<img file="MX368258B_D0066.tif" />

(S, E) -2,5-dimethyl-4- ((S) -N, 3,3-trimethyl-2 - ((S) -3-methyl acid

2- (methylamino) -3-phenylbutanamido) butanamido) hex-2-enóico (Compound F)

The title compound was synthesized using the methods described by Nieman et al. in J. Nat. Prod. 2003, 66, 183199.

Example 12

<img file="MX368258B_D0067.tif" />

Chemical formula: C₃₆H₅₄N₄OR₅S

Exact Mass: 654.38 (12) (S, E) -N- (mesylsulfonyl) -2,5-dimethyl-4 - ((S) -N, 3,3-trimethyl2 - ((S) -3-methyl-2 - (methylamino) -3phenylbutanamide) butanamide) hex-2-enamide

The title compound was prepared according to Example 3 and mesitylsulfonamide using General Procedures 2 and 7.

1 H NMR (400 MHz, Methanol-d4) δ 7.60 - 7.55 (m, 2H), 7.47 (m,

170

<td>2H),</td><td>7.37 (m,</td><td>1H), 7.03 (s, 2H), 6.50</td><td>(d, J = 6</td><td>Hz,</td><td>1 HOUR) ,</td><td> 5.06</td>

<td> - 4 .</td><td>91 (m, 3H</td><td>), 4.34 (s, 1H), 3.17</td><td>(s, 3H),</td><td> 2.68</td><td>(s,</td><td>6H),</td>

<td> 2.51</td><td>(s, 3H),</td><td>2.31 (s, 3H), 2.07 (m,</td><td>6.6 Hz,</td><td>2H),</td><td> 1.87</td><td>(s,</td>

<td>3H),</td><td>1.48 (s,</td><td>3H), 1.36 (s, 3H), 1.09</td><td>-1.04 (m,</td><td>J =</td><td> 16.8</td><td>Hz,</td>

10Η),

0.92 (t

J = 6.3 Hz, 6H).

C36H54N4O5S caled m / z = 654.38 found [M + H] + = 655.03

Example 13

<img file="MX368258B_D0068.tif" />

H

, .NH OR

O, zP

N

H

<img file="MX368258B_D0069.tif" />

Chemical formula:

Exact mass:

ocf₃

C34H47F3N4O6S

696.32 (13) (S, E) -2,5-dimethyl-N- (4- (trifluoromethoxy) phenylsulfonyl) -4 ((S) -N, 3,3-trimethyl-2 - ((S) -3- methyl-2- (methylamino) -3phenylbutanamido) butanamido) hex-2-enamide

The title compound is prepared according to Example 3 and the

4-trifluoromethoxyphenylsulfonamide using the

General Procedures 2 and 7.

Hz

1 HOUR)

4.95

1 HOUR) ,

1 H NMR (400 MHz, Methanol-d4) δ 8.16 (dd, J = 8.7, 1.4

<td> 7.69</td><td> - 7</td><td colspan="4">.28 (m, 4H), 6.52 (d, J = 9.2 Hz, 1H), 5.02 -</td>

<td>(m,</td><td>1 HOUR) ,</td><td>4.92 (s, OH), 4.35 (s, 1H),</td><td> 3.17</td><td>(s, 1H),</td><td> 2.51</td>

<td>1 HOUR) ,</td><td> 2.05</td><td>(ddd, J = 15.9, 10.9, 3.7</td><td>Hz,</td><td>1H), 1.87</td><td>(s,</td>

<td> 1.47</td><td>(s,</td><td>1H), 1.36 (s, 1H), 1.07 (s,</td><td>4H),</td><td>0.91 (t,</td><td>J =</td>

171

6.1 Ηζ, 3Η).

C34H47F3N4O6S caled m / z = 696.32 found [M + H] + = 697.26

Example 14

<img file="MX368258B_D0070.tif" />

Chemical formula: _C34H₅₀N₄OR₅S

Exact mass: 626.35 (14) (S, E) -N- (benzylsulfonyl) -2,5-dimethyl-4 - ((S) -N, 3,3-trimethyl-2 ((S) -3-methyl- 2- (methylamino) -3-phenylbutanamido) butanamide) hex2-enamide

The title compound was prepared according to Example 3 and benzylsulfonamide using General Procedures 2 and 7.

1 H NMR (400 MHz, Methanol-d4) δ 7.56 (d, J = 7.9 Ηζ, 2H), 7.47

<td>(t,</td><td>J = 7.3 Ηζ, 2H),</td><td> 7.38</td><td>(brs,</td><td>6H)</td><td colspan="2">, 6.39 (d, J =</td><td> 9.4</td><td>Hz,</td>

<td>1 HOUR) ,</td><td>5.06 (t, J = 10.0</td><td>Hz,</td><td>1H), 4</td><td> . 93</td><td>(s,</td><td>1H), 4.75</td><td>(s,</td><td>2H),</td>

<td> 4.36</td><td>(s, 1H), 3.13 (s,</td><td>3H),</td><td>2.51 i</td><td>: s,</td><td>3H),</td><td> 2.06-1.95</td><td>(m,</td><td>4H),</td>

<td> 1.48</td><td>(s, 3H), 1.39 (s,</td><td>3H),</td><td> 1.09</td><td>(s,</td><td>9H)</td><td>, 0.90 (t,</td><td>J =</td><td> 6.2</td>

Ηζ, 6H).

C34H47F3N4O6S caled m / z = 626.35 found [M + H] + = 626.99

172

Example 15

<img file="MX368258B_D0071.tif" />

Chemical formula: C₄₂H₅₆N₄OR₅S

Exact mass: 738.48 (15) (S, E) -2,5-dimethyl-N- (2,4,6triisopropylphenylsulfonyl) -4 - ((S) -N, 3,3-trimethyl-2 - ((S) -3methyl-2- (methylamino) -3-phenylbutanamido) butanamido) hex-2 enamide

The title compound was prepared according to Example 3 and 2,4,6-tri-isopropylphenylsulfonamide using them

General Procedures 2 and 7.

1 H NMR (400 MHz, Methanol-d4) δ 7.61 - 7.53 (m, 2H), 7.47 (t, J = 7.8 Hz, 2H), 7.41 - 7.33 (m, 1H), 7.27 (s, 2H), 6.50 ( dd,

J = 9.6, 1.8 Hz, 1H), 5.05 (t, J = 10.0 Hz, 1H), 4.92 (s,

1H), 4.43-4.26 (m, 3H), 3.16 (s, 3H), 2.94 (dd, J = 14.3,

7.4 Hz, 1H), 2.51 (s, 3H), 2.07 - 1.99 (m, 2H), 1.90 (d, J =

1.4 Hz, 3H), 1.48 (s, 4H), 1.39 (s, 3H), 1.33 - 1.22 (m,

18H), 1.11 (s, 2H), 1.06 (s, 9H), 0.91 (t, J = 6.0 Hz, 7H).

C42H66N4O5S caled m / z = 738.48 found [M + H] + = 738.10

173

Example 16

<img file="MX368258B_D0072.tif" />

Chemical formula: ru μ oc

Exact mass: 668.40 (16) (S, E) -N- (4-tert-butylphenylsulfonyl) -2,5-dimethyl-4 - ((S) -N, 3,3-trimethyl-2 - ((S) -3 -methyl-2- (methylamino) -3phenylbutanamido) butanamido) hex-2-enamide

The title compound was prepared according to Example 3 and the

4-tertbutylphenylsulfonamide using the Procedures

General 2 and 7.

<td>1 HOUR</td><td>NMR (400</td><td colspan="2">MHz, Methanol-d4) δ 7.98 (d,</td><td>J =</td><td>8.6 Hz, 2H)</td><td>t</td><td> 7.64</td>

<td>(d,</td><td>J = 8.6</td><td>Hz, 2H</td><td>), 7.55 (d, J = 7.9</td><td>Hz,</td><td>2H), 7.47 (</td><td>t</td><td>J =</td>

<td> 7.7</td><td>Hz, 3H),</td><td> . 7.37 (</td><td>t, J = 7.1 Hz, 1H),</td><td> 6.48</td><td>(dd, J = 9.</td><td> 6,</td><td> 1.8</td>

<td>Hz,</td><td>1H), 4.</td><td>99 (t,</td><td>J = 10.0 Hz, 1H), 4</td><td> . 92</td><td>(s, 1H), 4.</td><td> 35</td><td>(s,</td>

<td>1 HOUR)</td><td> , 3.16 (</td><td>s, 3H),</td><td>2.51 (s, 3H), 1.87</td><td>(d,</td><td>J = 1.4 Hz</td><td>F</td><td>3H),</td>

<td> 1.4</td><td>7 (s, 3H</td><td> ), 1.38</td><td>(s, 10H), 1.06 (s,</td><td>9H),</td><td>0.91 (t, J</td><td></td><td> 6.2</td>

Hz, 7H).

C42H66N4O5S caled m / z = 668.40 found [M + H] + = 669.28

174

Example 17

<img file="MX368258B_D0073.tif" />

Chemical formula: C₃₃H₄₇CIN₄OR_sS

Exact mass: 646.30 (17) (S, E) -N- (4-chlorophenylsulfonyl) -2,5-dimethyl-4 - ((S) -N, 3,3-trimethyl-2 - ((S) -3-methyl -2- (methylamino) -3phenylbutanamido) butanamide) hex-2-enamide

The title compound was prepared according to Example 3 and the

4-chlorophenylsulfonamide using the Procedures

General 2 and 7.

<td colspan="2">1 H NMR</td><td colspan="3">(400 MHz, Methanol-d4)</td><td>) δ 8</td><td>.03 (d,</td><td>J =</td><td> 8.7</td><td>Hz, 2H),</td><td> 7.60</td>

<td>(d,</td><td>J =</td><td>8.7 Hz</td><td>, 2h:</td><td> ), 7.57 -</td><td> 7.51</td><td>(m, 2H</td><td> ), 7</td><td> .47</td><td>(dd, J =</td><td> 8.6,</td>

<td> 6.9</td><td>Hz,</td><td>2H), 7</td><td> .42 -</td><td>- 7.32 (m</td><td>., 1 HOUR)</td><td> , 6.50</td><td>(dd,</td><td>J =</td><td> =9.2, 1.7</td><td>Hz,</td>

<td>1 HOUR)</td><td> , 4.</td><td>96 (dd,</td><td>J =</td><td> 10.9, 9.</td><td>1 Hz,</td><td>2H), 4</td><td> . 92</td><td>(s,</td><td>1H), 4.35</td><td>(s,</td>

<td>1 HOUR)</td><td> , 3.</td><td>17 (s,</td><td>3H),</td><td>2.51 (s</td><td>, 3H)</td><td> , 2.14</td><td> - 2.</td><td> 03</td><td>(m, 1H),</td><td> 2.01</td>

<td>(s,</td><td>1 HOUR)</td><td> , 1.87</td><td>(d,</td><td>J = 1.4</td><td>Hz,</td><td>3H), 1.</td><td> 46 (</td><td>s,</td><td>3H), 1.36</td><td>(s,</td>

3H), 1.07 (s, 9H), 0.91 (dd, J = 6.5, 4.6 Hz, 7H).

C33H47C1N4O5S caled m / z = 646.30 found [M + H] + = 647.20

175

Example 18

<img file="MX368258B_D0074.tif" />

Chemical formula: _C34H₄₇N₅OR₃S

Exact mass: 637.33 (18) (S, E) -N- (3-cyanophenylsulfonyl) -2,5-dimethyl-4 - ((S) -N, 3, 3-trimethyl-2 - ((S) -3-methyl -2- (methylamino) -3phenylbutanamido) butanamide) hex-2-enamide

The title compound was prepared according to Example 3 and 3-cyanophenylsulfonamide using General Procedures 2 and 7.

1 H NMR (400 MHz, Methanol-d4) δ 8.38 (s, 1 H), 8.31 (dt, J =

<td> 8.0,</td><td>1.5 Hz, 1H),</td><td> 8.02 - 7.</td><td> 92 (</td><td>m, 1H), 7.75 (t</td><td>, J = 7.9</td><td>Hz,</td>

<td>1 HOUR) ,</td><td>7.53 (d, J =</td><td>^: 1.2 Hz,</td><td>1 HOUR) ,</td><td>7.48 (dd, J =</td><td> 8.6, 6.9</td><td>Hz,</td>

<td>2H),</td><td> 7.43 - 7.33</td><td>(m, 1H),</td><td> 6.55</td><td>(dd, J = 9.3,</td><td>1.7 Hz,</td><td>1 HOUR) ,</td>

<td> 4.93</td><td>(d, J = 5.4</td><td>Hz, 2H),</td><td> 4.35</td><td>(s, 1H), 3.18</td><td>(s, 3H),</td><td> 2.51</td>

<td>(s,</td><td>3H), 2.15-1.</td><td>98 (m, 2H</td><td>), i.</td><td>.87 (d, J = 1.4</td><td>Hz, 3H),</td><td> 1.45</td>

<td>(s,</td><td>3H), 1.32 (s,</td><td>3H), 1.07</td><td>(s,</td><td>9H), 0.92 (dd,</td><td>J = 6.6,</td><td> 3.9</td>

<td>Hz,</td><td>7H).</td><td></td><td></td><td></td><td></td><td></td>

C34H47N5O5S caled m / z = 637.33 found [M + H] + = 638.00

176

Example 19

Chemical formula: c₃₃H₄₇N₅OR₇S

Exact mass: 657.32 (19) (S, E) -2,5-dimethyl-N- (2-nitrophenylsulfonyl) -4 - ((S) -N, 3, 3 trimethyl-2 - ((S) -3- methyl-2- (methylamino) -3 phenylbutanamido) butanamido) hex-2-enamide

The title compound was prepared according to

Example 3 and the

2-nitrophenylsulfonamide using the

Procedures

General 2 and 7.

1 H NMR (400 MHz, Methanol-d4) δ 8.36 - 8.27 (m,

1H), 7.82 (dd,

J = 5.9, 3.8 Hz, 3H), 7.61 - 7.51 (m, 2H), 7.47 (dd,

J = 8.6,

6.9 Hz, 2H), 7.42

7.31 (m, 1H), 6.63 (dd, J = 9.5,

1.7 Hz,

1H), 5.03 (t, J

10.0 Hz,

1H), 4.93 (s, 1H), 4.36 (s, 1H),

3.18 (s,

3H), 2.51 (s, 3H),

2.12 - 2.01 (m, 1H), 1.88 (d, J

1.4 Hz,

3H), 1.48 (s, 3H),

1.37 (s, 3H), 1.06 (s, 9H), 0.97

0.86 (m,

6H).

C34H47N5O5S caled m / z = 657.32 found [M + H] + = 658.21

177

Example 20

<img file="MX368258B_D0075.tif" />

Chemical formula: C₃₄H₄₉N₅OR₈S Exact mass: 687.33 (20) (S, E)^-N- (4-methoxy-2-nitrophenylsulfonyl) -2,5-dimethyl-4 - ((S) N, 3,3-trimethyl-2 - ((S) -3-methyl-2- (methylamino) -3-phenylbutanamido ) butanamide) hex-2-enamide

The title compound was prepared according to Example 3 and the

2-nitro-4-methoxyphenylsulfonamide using the

<td colspan="2">General Procedures 2 and</td><td> 7 .</td><td></td><td></td><td></td><td></td><td></td>

<td colspan="2">1 H NMR (400 MHz, Methanol-d4)</td><td>δ 8.</td><td>24 (d,</td><td>J = 8.9</td><td>Hz,</td><td>1 HOUR) ,</td><td> 7.59</td>

<td> - 7.</td><td>51 (m, 2H), 7.47 (t, J</td><td> = 7.</td><td>.6 Hz,</td><td>2H), 7.</td><td> 44 -</td><td> 7.25</td><td>(m,</td>

<td>4H),</td><td>6.60 (dd, J = 9.2, 1.7</td><td>Hz,</td><td>1H), 5</td><td>.03 (t,</td><td>J =</td><td> 10.0</td><td>Hz,</td>

<td>1 HOUR) ,</td><td>4.93 (s, 1H), 4.36 (s,</td><td>1 HOUR) ,</td><td> 3.97 (</td><td>s, 3H),</td><td> 3.18</td><td>(s,</td><td>3H),</td>

<td> 2.51</td><td>(s, 3H), 2.13-2.02 (m</td><td>. ih;</td><td> ) , 1.89</td><td>(d, J =</td><td> = 1.4</td><td>Hz,</td><td>3H),</td>

<td> 1.48</td><td>(s, 3H), 1.38 (s, 3H),</td><td> 1.11</td><td>(s, 2H</td><td> ), 1.06</td><td>(s,</td><td>9H),</td><td> 0.99</td>

- 0.88 (m, 6H).

C34H49N5O8S caled m / z = 687.33 found [M + H] + = 689.23

178

Example 21

<img file="MX368258B_D0076.tif" />

nh₂

Chemical formula: C34H₄₈N₆OR₈S

Exact mass: 700.33 (21)

4- (N - ((S, E) -2,5-dimethyl-4 - ((S) -N, 3,3-trimethyl-2 - ((S) -3-methyl2- (methylamino) -3- phenylbutanamide) butanamide) hex-2enoyl) sulfamoyl) -3-nitrobenzamide

The title compound was prepared according to Example 3 and the

3-nitro-4-sulfamoylbenzamide using General Procedures 2 and 7.

1H NMR (400 MHz, Methanol-d4) δ 8.35 (d, J = 8.0 Hz, 1H), 8.22

<td>(d,</td><td>J =</td><td colspan="2">8.0 Hz,</td><td>2H),</td><td> 7.59 - 7.51</td><td>(m, 2H)</td><td> , 7.4</td><td>7 (t</td><td>, J =</td><td> 7.6</td>

<td>Hz,</td><td>2H),</td><td> 7.37</td><td>(t,</td><td>J =</td><td>7.3 Hz, 1H),</td><td> 6.70 -</td><td> 6.57</td><td>(m,</td><td>1 HOUR) ,</td><td> 5.04</td>

<td>(t,</td><td>J -</td><td> 10.0</td><td>Hz,</td><td>1 HOUR) ,</td><td>4.94 (s, 1 H</td><td> ), 4.37</td><td>(s,</td><td>1 HOUR) ,</td><td> 3.17</td><td>(s,</td>

<td>3H),</td><td colspan="2">2.52 (s,</td><td>3H</td><td> ), 2.</td><td>05 (ddd, J =</td><td> = 10.3,</td><td> 7.4,</td><td> 5.5</td><td>Hz,</td><td>1 HOUR) ,</td>

<td> 1.87</td><td>(d,</td><td>J =</td><td> 1.4</td><td>Hz,</td><td>3H), 1.48 (s</td><td>, 3H),</td><td> 1.38</td><td>(s,</td><td>3H),</td><td> 1.06</td>

<td>(s,</td><td>9H),</td><td> 0.92</td><td>(dd</td><td>, J =</td><td>14.7, 6.8 Hz</td><td>, 6H).</td><td></td><td></td><td></td><td></td>

C34H48N6O8S caled m / z = 700.33 found [M + H] + = 701.28

179

Example 22

<img file="MX368258B_D0077.tif" />

NH

C ^ soN ^ S

642.35

Chemical formula: Exact mass:

(22) (S, E) -N- (4-methoxyphenylsulfonyl) -2,5-dimethyl-4 - ((S) -N, 3,3 trimethyl-2 - ((S) -3-methyl-2- (methylamino) -3 phenylbutanamide) butanamide) hex-2-enamide

The title compound was prepared according to Example 3 and the

4-methoxyphenylsulfonamide using

General 2 and 7.

the procedures

1 H NMR (400

9.0

7.54

MHz, Methanol-d4) δ 7.97 (d,

Hz, 2H)

<td>(d,</td><td colspan="2">J = 7.5 Hz, 2H),</td><td> 7.46</td><td>(t, J = 7.6</td><td>Hz,</td><td>2H), 7.36 (t,</td><td>J =</td>

<td> 7.2</td><td>Hz, 1H), 7.06</td><td>(d,</td><td>J =</td><td>9.0 Hz, 2H),</td><td> 6.48</td><td>(dd, J = 9.3,</td><td> 1.9</td>

<td>Hz,</td><td>1H), 4.97 (t,</td><td>J</td><td> = 9.</td><td>9 Hz, 1H), 4</td><td> . 92</td><td>(s, 1H), 4.22</td><td>(s,</td>

<td>1 HOUR) ,</td><td>3.89 (s, 3H)</td><td> , 3</td><td> . 15</td><td>(s, 3H), 2.46</td><td>(s,</td><td>3H), 2.10 -</td><td> 1.99</td>

<td>(m,</td><td>2H), 1.86 (d,</td><td>J</td><td> = 1.</td><td>4 Hz, 3H), 1</td><td> .46</td><td>(s, 3H), 1.36</td><td>(s,</td>

<td>3H),</td><td>1.06 (s, 9H),</td><td> 0.</td><td> 94 -</td><td>0.84 (m, 6H).</td><td></td><td></td><td></td>

C34H50N4O6S caled m / z = 642.35 found [M + H] + = 643.31

180

Example 23

<img file="MX368258B_D0078.tif" />

Chemical formula: c₃₅H₄₈F₃N₅OR₆S

Exact mass: 723.33 (23) (S, E) -2,5-dimethyl-N- (4- (2,2,2trifluoroacetamido) phenylsulfonyl) -4 - ((S) -N, 3,3-trimethyl-2 ((S) -3-methyl-2- (methylamino) -3-phenylbutanamido) butanamido) hex2-enamide

The title compound was prepared according to Example 3 and the

2,2,2-trifluoro-N- (4-sulfamoylphenyl) acetamide using the

General Procedures 2 and 7.

1 H NMR (400 MHz, Methanol-d4) δ 8.06 (d, J = 8.9 Hz, 2H), 7.88 (d, J = 8.9 Hz, 2H), 7.52 (d, J = 7.1 Hz, 2H), 7.49 - 7.40 (m, 3H), 7.35 (dd, J = 8.1, 6.1 Hz, 1H), 6.47 (dd, J = 9.2, 1.8 Hz, 1H), 4.33 (s, 1H), 3.15 (s, 3H), 2.48 ( s, 3H), 2.13 1.96 (m, 2H), 1.85 (d, J = 1.4 Hz, 3H), 1.43 (s, 3H), 1.33 (s, 3H), 1.04 (s, 9H), 0.89 (dd, J = 6.8, 4.7 Hz, 6H).

C35H48F3N5O6S caled m / z = 723.33 found [M + H] + = 724.08

181

Example 24

OR_{x Z}OR

<img file="MX368258B_D0079.tif" />

, NH

NH₂

Chemical formula: C₃₃H₄₉N₅OR₅S

Exact mass: 627.35 (24) (S, E) -N- (4-aminophenylsulfonyl) -2,5-dimethyl-4 - ((S) -N, 3,3 trimethyl-2 - ((S) -3- methyl-2- (methylamino) -3 phenylbutanamido) butanamido) hex-2-enamide

The title compound was prepared according to Example 3

2,2,2-trifluoro-N- (4-sulfamoylphenyl) acetamide using and the

General Procedures 2, 3 and 7.

<td>^XH</td><td>NMR</td><td>(400 MHz, Methanol-d4)</td><td>δ 7.71</td><td>(d,</td><td>J =</td><td> 8.8</td><td>Hz, 2H),</td><td> 7.55</td>

<td>(d,</td><td>J =</td><td>; 7.6 Hz, 2H), 7.47 (</td><td>d, J = 6</td><td> . 9</td><td>Hz,</td><td>2H),</td><td>7.37 (t,</td><td>J =</td>

<td> 6.8</td><td>Hz,</td><td>1H), 6.67 (d, J = 8.</td><td>8 Hz, 2H</td><td> ) ,</td><td> 6.44</td><td>(dd</td><td>, J = 9.2,</td><td> 1.6</td>

<td>Hz,</td><td>1 HOUR)</td><td>, 4.97 (t, J = 9.7</td><td>Hz, 1H),</td><td> 4</td><td> . 92</td><td>(s,</td><td>1H), 4.36</td><td>(s,</td>

<td>1 HOUR)</td><td> , 3.</td><td>16 (s, 3H), 2.51 (s,</td><td>3H), 2,</td><td> .16</td><td> - 2</td><td> .00</td><td>(m, 1H),</td><td> 1.87</td>

<td>(d,</td><td>J =</td><td>= 1.4 Hz, 3H), 1.46</td><td>(s, 3H),</td><td> 1</td><td> . 37</td><td>(s,</td><td>3H), 1.07</td><td>(s,</td>

<td>9H)</td><td>, or.</td><td>92 (d, J = 6.4 Hz, 3H</td><td> ) , 0.91</td><td>(d,</td><td>J =</td><td> 6.3</td><td>Hz, 3H).</td><td></td>

C33H49N5O5S caled m / z = 627.35 found [M + H] + = 628.35

182

Example 25

<img file="MX368258B_D0080.tif" />

Chemical formula: Ο₃₃Η₄₈Ν₄Ο₅8

Exact mass: 612.33 (25) (S, E) -2,5-dimethyl-N- (phenylsulfonyl) -4 - ((S) -N, 3,3-trimethyl-2 ((S) -3- methyl-2- (methylamino) -3-phenylbutanamido) butanamido) hex

2-enamide

The title compound was prepared according to Example 3 and phenylsulfonamide using General Procedures 2 and

7.

1 H NMR (400 MHz, Methanol-d4) δ 8.06 - 7.95 (m, 2H), 7.63 15 7.40 (m, 8H), 7.40 - 7.30 (m, 1H), 6.53 (dd, J = 9.3, 1.6 Hz,

<td>1H), 5</td><td> .05 - 4.95</td><td>(m,</td><td>1 HOUR) ,</td><td> 4.22</td><td>(s, 1H),</td><td> 3.14</td><td>(s,</td><td>3H)</td><td> , 2.45</td>

<td>(s, 3H:</td><td> 1, 2.09 - 1.</td><td> 95</td><td colspan="2">(m, 1H), 1.</td><td>85 (d, J</td><td> = 1.4</td><td>Hz,</td><td>3H)</td><td> , 1.46</td>

<td>(s, 3H)</td><td>i, 1.36 (s,</td><td>3H)</td><td> , 1.06</td><td>(s,</td><td>9H), 0.89</td><td>(dd,</td><td>J =</td><td> 11.</td><td> 9, 6.5</td>

Hz, 7H).

C33H48N4O5S caled m / z = 612.33 found [M + H] + = 613.06

183

<img file="MX368258B_D0081.tif" />

<img file="MX368258B_D0082.tif" />

Example 26

<img file="MX368258B_D0083.tif" />

Ύ H go '

NH O ...

Chemical formula: C₃₄H_SWFN₅0₅S

Exact mass: 659.35 (26) (S, E) -N- (N- (2-fluorobenzyl) sulfamoyl) -2,5-dimethyl-4- ((S) -

N, 3,3-trimethyl-2 - ((S) -3-methyl-2- (methylamino) -3phenylbutanamido) butanamido) hex-2-enamide

2-Fluorobenzylsulfamamide was prepared from

2fluorobenzylamine according to the General Procedure

14;

The title compound was prepared according to Example 3 and the

2fluorobenzyl sulfamamide using the Procedures

<td>Generale</td><td>s 2 and</td><td> 7 .</td><td></td><td></td><td></td><td></td>

<td>1 H NMR</td><td colspan="2">(400 MHz, Methanol-d4</td><td>) δ 7.</td><td>63 - 7.41 (m, 6H</td><td> ), 7.41</td><td> —</td>

<td>7.26 (m,</td><td>3H),</td><td>7.14 (td, J =</td><td> 7.5, 1</td><td>.2 Hz, 1H), 7.07</td><td>(ddd, J</td><td> =</td>

<td> 9.5, 8.2</td><td> , 1.1</td><td>Hz, 1H), 6.37</td><td>(dd, J</td><td>= 9.4, 1.7 Hz, 1H</td><td> :), 5.07</td><td> —</td>

<td>4.97 (m,</td><td>1 HOUR) ,</td><td>4.37 (s, 1 H),</td><td> 4.33 (</td><td>s, 2H), 3.15 (s,</td><td>3H), 2.</td><td> 51</td>

<td>(s, 3H),</td><td> 2.10</td><td>- 1.97 (m, 1H)</td><td> , 1.83</td><td>(d, J = 1.4 Hz,</td><td>3H), 1.</td><td> 49</td>

<td>(s, 3H),</td><td> 1.39</td><td>(s, 3H), 1.09</td><td>(s, 9H)</td><td>, 0.97 - 0.84 (m,</td><td>6H).</td><td></td>

C34H50FN5O5S caled m / z = 659.35 found [M + H] + = 660.28

184

Example 27

<img file="MX368258B_D0084.tif" />

Chemical formula: c₃₂H₅₃N₅OR₅S

Exact mass: 619.38 (27) (S, E) -2,5-dimethyl-N- (piperidin-1-ylsulfonyl) -4 - ((S) -N, 3,3trimethyl-2 - ((S) -3 -methyl-2- (methylamino) -3phenylbutanamido) butanamido) hex-2-enamide

Piperidin-l-sulfonamide was synthesized from piperidine according to General Procedure 14; The title compound was prepared according to Example 3 and piperidin-l-sulfonamide using the Procedures

<td colspan="7">General 2 and 7.</td>

<td colspan="2">1 H NMR (400 MHz, Methanol-d4)</td><td>δ 7.55</td><td>(d, J = 1.2</td><td>Hz,</td><td>1 HOUR) ,</td><td> 7.47</td>

<td>(t, J = 7.6 H</td><td>z, 3H), 7.42 -</td><td>7.29 (m,</td><td>1H), 6.48</td><td>(dd_z</td><td>J =</td><td> 9.7,</td>

<td>1.8 Hz, 1H),</td><td>5.05 (t, J =</td><td>10.0 Hz,</td><td>1H), 4.39</td><td>(s,</td><td>1 HOUR) ,</td><td> 3.18</td>

<td>(s, 3H), 2.52</td><td>(s, 3H), 2.07</td><td>(d, J =</td><td colspan="2">10.5 Hz, 1H), 1</td><td> . 96</td><td>(d, J</td>

<td>= 1.4 Hz, 3H)</td><td colspan="2">, 1.61 (ddd, J = 20.0,</td><td> 10.3, 5.4</td><td>Hz,</td><td>9H),</td><td> 1.49</td>

<td>(s, 4H), 1.39</td><td>(s, 3H), 1.09</td><td>(s, 9H),</td><td> 0.99 - 0.84</td><td>(m,</td><td>9H).</td><td></td>

C32H53N5O5S caled m / z = 619.38 found [M + H] + = 620.38

185

Example 28

<img file="MX368258B_D0085.tif" />

Chemical formula:

Exact mass: 626.35 (28) (S, E) -2,5-dimethyl-N- (o-tolylsulfonyl) -4 - ((S) -N, 3,3-trimethyl2 - ((S) -3-methyl -2- (methylamino) -3phenylbutanamido) butanamide) hex-2-enamide

The title compound was prepared according to Example 3 and the

2-toluenesulfonamide using General Procedures 2 and 7.

<td colspan="3">1 H NMR (4 00 MHz,</td><td>Methanol-d4</td><td>) δ</td><td> 8.10</td><td>(dd, J =</td><td colspan="2"> 8.0, 1.4</td><td>Hz,</td>

<td>1 HOUR) ,</td><td> 7.60</td><td> - 7.33</td><td>(m, 11H),</td><td> 6.52</td><td>(dd,</td><td>, J = 9.6,</td><td> 1.7</td><td>Hz,</td><td>1 HOUR) ,</td>

<td> 5.04</td><td> - 4.</td><td>90 (m,</td><td>2H), 4.35</td><td>(s,</td><td>1 HOUR) ,</td><td>3.18 (s,</td><td>3H),</td><td> 2.67</td><td>(s,</td>

<td>3H),</td><td> 2.51</td><td>(s, 3H)</td><td> , 2.15 - 2</td><td> .03</td><td>(m,</td><td>2H), 2.01</td><td>(s,</td><td>1 HOUR) ,</td><td> 1.87</td>

<td>(d,</td><td>J = 1</td><td>. 4 Hz,</td><td>3H), 1.46</td><td>(s,</td><td>3H),</td><td>1.35 (s,</td><td>3H),</td><td> 1.07</td><td>(s,</td>

<td>9H),</td><td> 0.92</td><td>(t, J =</td><td>6.3 Hz, 6H)</td><td></td><td></td><td></td><td></td><td></td><td></td>

C34H50N4O5S caled m / z = 626.35 found [M + H] + = 627.05

186

Example 29

<img file="MX368258B_D0086.tif" />

Chemical formula: C₃₃H4₇Brn₄OR_sS

Exact mass: 690.25 (29) (S, E) -N- (4-bromophenylsulfonyl) -2,5-dimethyl-4 - ((S) -N, 3,3-trimethyl-2 - ((S) -3-methyl -2- (methylamino) -310 phenylbutanamido) butanamide) hex-2-enamide

The title compound was prepared according to Example 3 and the

4-Bromophenylsulfonamide using the Procedures

General 2 and 7.

1 H NMR (400 MHz, Methanol-d4) δ 7.95 (d, J = 8.3 Hz, 2H), 7.76

<td> 15</td><td colspan="2">(d, J = 8.0 Hz, 2H</td><td colspan="2"> ), 7.55</td><td>(d, J</td><td> = 7.5</td><td>Hz, 2H), 7.47</td><td>(dd,</td><td>J =</td>

<td></td><td>8.6, 6.9 Hz,</td><td>2H),</td><td> 7.41</td><td> -</td><td colspan="2">7.29 (m, 1 H),</td><td>6.51 (d, J =</td><td> 9.0</td><td>Hz,</td>

<td></td><td>1H), 4.35 (s,</td><td>1 HOUR) ,</td><td> 3.16</td><td> (</td><td>s, 3H),</td><td> 2.50</td><td>(s, 3H), 2.06</td><td>(dt,</td><td>J =</td>

<td></td><td>10.7, 6.3 Hz,</td><td>1 HOUR) ,</td><td> 1.87</td><td> (</td><td>s, 3H),</td><td> 1.46</td><td>(s, 3H), 1.36</td><td>(s,</td><td>3H),</td>

<td></td><td>1.07 (s, 9H),</td><td> 0.91</td><td>(dd,</td><td>J</td><td> = 6.9,</td><td>Four . 9 Hz</td><td>, 8H).</td><td></td><td></td>

<td> 20</td><td>C33H47BrN4O5S</td><td colspan="2">caled m / z</td><td> =</td><td> 690.25</td><td colspan="2">found [M + H] + =</td><td> 691</td><td> .17,</td>

693.18

187

Example 30

<img file="MX368258B_D0087.tif" />

Chemical formula: c₃₇H₅₀N₄OR₅S

Exact mass: 662.35 (30) (S, E) -2,5-dimethyl-N- (naphthalen-2-ylsulfonyl) -4 - ((S) -N, 3,3trimethyl-2 - ((S) -3 -methyl-2- (methylamino) -3phenylbutanamido) butanamido) hex-2-enamide

The title compound was prepared according to Example 3 and the

2-Naphthylsulfonamide using General Procedures 2 and 7.

1 H NMR (400 MHz, Methanol-d4) δ 8.69 - 8.62 (m, 1 H), 8.47 (d,

J = 8.2 Hz, 1H), 8.14 - 7.95 (m, 5H), 7.71 (dddd, J = 18.4,

<td> 8.2,</td><td> 6.9</td><td>i</td><td> . 4</td><td>Hz, 2H),</td><td> 7.57</td><td> - 7.50</td><td>(m, 2H), 7.46</td><td>(dd,</td><td>J =</td>

<td> 8.6,</td><td> 6.9</td><td>Hz,</td><td colspan="2">2H), 7.42</td><td> - 7.33</td><td>(m, 1H)</td><td>, 6.50 (dd, J =</td><td colspan="2"> 9.3, 1.5</td>

<td>Hz,</td><td>1 HOUR) ,</td><td> 4.</td><td> 92</td><td> - 4.87</td><td>(m, 1H)</td><td> , 4.34</td><td>(s, 1H), 3.16</td><td>(s,</td><td>3H),</td>

<td> 2.50</td><td>(s,</td><td>3H)</td><td>t</td><td> 2.13 - 1.</td><td>99 (m,</td><td>1H), 1</td><td>.85 (d, J = 1.4</td><td>Hz,</td><td>3H),</td>

<td> 1.44</td><td>(s,</td><td>3H)</td><td>F</td><td>1.34 (s,</td><td>3H), 1</td><td>.04 (s,</td><td>9H), 0.90 (dd,</td><td colspan="2">J = 6.6,</td>

4.0 Hz, 6H).

C37H50N4O5S caled m / z = 662.35 found [M + H] + = 663.32

188

Example 31

<img file="MX368258B_D0088.tif" />

OMe

Chemical formula: c₃₅H₅₀N₄OR₇S

Exact mass: 670.34 (31)

Methyl 4- (N- ((S, E) -2,5-dimethyl-4 - ((S) -N, 3,3-trimethyl-2 - ((S) -3methyl-2- (methylamino) -3 -phenylbutanamido) butanamido) hex-2enoyl) sulfamoyl) benzoate

The title compound was prepared according to Example 3 and 4-carboxymethylphenylsulfonamide using General Procedures 2 and 7.

1 H NMR (400 MHz, Methanol-d4) δ 8.24 - 8.10 (m, 4H), 7.58 7.50 (m, 2H), 7.47 (dd, J = 8.6, 6.9 Hz, 2H), 7.41 - 7.33 (m, 1H) , 6.52 (dd, J = 9.2, 1.6 Hz, 1H), 4.35 (s, 1H), 3.97 (s, 3H), 3.18 (s, 3H), 2.50 (s, 3H), 2.15 - 2.00 (m, 1H) ), 1.86 (d, J = 1.4 Hz, 3H), 1.45 (s, 3H), 1.35 (s, 3H), 1.07 (s, 9H), 0.91 (dd, J = 6.7, 3.8 Hz, 6H).

C35H50N4O7S caled m / z = 670.34 found [M + H] + = 671.10

189

Example 32

<img file="MX368258B_D0089.tif" />

Chemical formula: C35H50F3N5O5S

Exact mass: 709.35 (32) (S, E) -2,5-dimethyl-N- (N- (2- (trifluoromethyl) benzyl) sulfamoyl) 4 - ((S) -N, 3,3-trimethyl-2 - ((S) -3-methyl-2- (methylamino) -3phenylbutanamido) butanamido) hex-2-enamide

The title compound was prepared according to Example 3 and 2-trifluoromethylbenzyl sulfonamide using the

General Procedures 2 and 7.

NMR (400 MHz, Methanol-d4) δ 7.78 (d, J = 7.9 Hz, 1H), 7.74

<td> -7.67 (</td><td>my h:</td><td> ), 7.</td><td> 64 (</td><td>Dd, J = 8.1, 6.7 Hz, 1H)</td><td> , 7.60 -</td><td> 7.52</td>

<td>(m, 2H),</td><td> 7.48</td><td>(dd,</td><td>J =</td><td>8.5, 6.8 Hz, 4H), 7.42-</td><td>7.33 (m,</td><td>1 HOUR) ,</td>

<td> 6.48 - 6</td><td colspan="2">.40 (m, 1H</td><td> ), 5</td><td>.11 - 5.02 (m, 1H), 4.45</td><td>(s, 2H),</td><td> 4.37</td>

<td>(s, 1H),</td><td> 3.17</td><td>(s,</td><td>3H)</td><td>, 2.52 (s, 3H), 2.11 -</td><td>1.99 (m,</td><td>2H),</td>

<td>1.92 (d,</td><td>J =</td><td> 1.4 '</td><td>Hz,</td><td>3H), 1.49 (s, 3H), 1.40</td><td>(s, 3H),</td><td> 1.09</td>

<td>(s, 9H),</td><td> 0.92</td><td>(dd,</td><td>J =</td><td>9.3, 6.7 Hz, 6H).</td><td></td><td></td>

C35H50F3N5O5S caled m / z = 709.35 found [M + H] + = 710.02

190

Example 33

<img file="MX368258B_D0090.tif" />

Chemical formula: C3₃H₅₆N4O₅S

Exact mass: 620.40 (33) (4S, E) -N- (hexan-2-ylsulfonyl) -2,5-dimethyl-4 - ((S) -N, 3,3-trimethyl-2 - ((S) -3 -methyl-2- (methylamine) -3phenylbutanamide) butanamide) hex-2-enamide

The title compound was prepared according to Example 3 and hexane-2-sulfonamide using General Procedures 2 and 7.

1H NMR (400 MHz, Methanol-d4) δ 7.56 - 7.48 (m, 2H), 7.42 (t, J = 7.8 Hz, 2H), 7.31 (t, J = 7.3 Hz, 1H), 6.58 - 6.50 (m, 1H), 5.05 (t, J = 10.0 Hz, 1H), 4.92 (s, 1H), 3.84 (s, 1H), 3.65 (dt, J = 10.8, 4.3 Hz, 1H), 3.14 (s, 3H), 2.32 (s, 3H), 2.09 - 1.96 (m, 2H), 1.93 (d, J = 1.4 Hz, 3H), 1.61 - 1.27 (m, 3H), 1.06 (s, 9H), 0.98 - 0.90 (m, 6H), 0.87 (d, J = 6.5 Hz, 3H).

C33H56N4O5S caled m / z = 620.40 found [M + H] + = 621.55

191

Example 34

<img file="MX368258B_D0091.tif" />

Chemical formula: CgoHgo ^ OgS

Exact mass: 594.35 (34) (S, E) -N- (2-methoxyethylsulfonyl) -2,5-dimethyl-4 - ((S) -N, 3,3-trimethyl-2 - ((S) -3-methyl -2- (methylamino) -3phenylbutanamido) butanamide) hex-2-enamide

The title compound was prepared according to Example 3 and the

2-Methoxyethanesulfonamide using the Procedures

General 2 and 7.

1H NMR (400 MHz, Methanol-d4) δ 7.56 (d, J = 7.8 Hz, 2H), 7.47 (t, J = 7.6 Hz, 2H), 7.37 (t, J = 7.3 Hz, 1H), 6.51 (d , J =

<td> 9.4</td><td>Hz,</td><td>1 HOUR)</td><td> /</td><td> 5.07</td><td>(t,</td><td>J</td><td> = 10.0</td><td>Hz, 1H)</td><td>, 4.95 i</td><td>ÍS, 1H),</td><td> 4.33</td>

<td>(s,</td><td>1 HOUR) ,</td><td> 3.</td><td> 82</td><td>(t,</td><td>J =</td><td> 5.8</td><td>Hz, 2H</td><td> ), 3.70</td><td>(q, J =</td><td>5.2 Hz,</td><td>2H),</td>

<td> 3.18</td><td colspan="2">(s, 3H</td><td> ) _r</td><td> 2.50</td><td>(s,</td><td>3H)</td><td> , 2.18</td><td> - 2.00</td><td>(m, 1H),</td><td>1.95 (d,</td><td>J =</td>

<td> 1.4</td><td>Hz,</td><td>3H)</td><td>F</td><td> 1.49</td><td>(s,</td><td>3H)</td><td> , 1.39</td><td>(s, 3H)</td><td> , 1.09</td><td>(s, 9H),</td><td> 0.93</td>

<td>(dd,</td><td colspan="2">J = 14</td><td> • 8,</td><td> 6.6</td><td>Hz,</td><td>6H)</td><td></td><td></td><td></td><td></td><td></td>

C30H50N406S caled m / z = 594.35 found [M + H] + = 595.44

192

Example 35

<img file="MX368258B_D0092.tif" />

Chemical formula: C ^ H ^^ OgS

Exact mass: 618.38 (35) (S, E) -N- (cyclopentylmethylsulfonyl) -2,5-dimethyl-4 - ((S) -N, 3,3-trimethyl-2 - ((S) -3-methyl-2 - (methylamino) -3phenylbutanamide) butanamide) hex-2-enamide

The title compound was prepared according to Example 3 and cyclopentylmethanesulfonamide using General Procedures 2 and 7.

1 H NMR (400 MHz, Methanol-d4) δ 7.61 - 7.52 (m, 2H), 7.48 (dd,

<td>J = 8.6,</td><td colspan="2">6.9 Hz,</td><td colspan="2">2H), 7.38</td><td colspan="2">(t, J = 7.4 Hz,</td><td>1 HOUR) ,</td><td> 6.54</td><td>(dd,</td><td>J =</td>

<td> 9.4, 1.7</td><td>Hz,</td><td>1 HOUR</td><td> ), 5.06</td><td>(t</td><td>, J = 10.0</td><td>Hz,</td><td>1 HOUR) ,</td><td> 4.94</td><td>(s,</td><td>1 HOUR) ,</td>

<td>4.37 (s,</td><td>1 HOUR)</td><td> , 3.</td><td>52 (dd,</td><td>J</td><td> = 7.0, 5.4</td><td>Hz,</td><td>3H),</td><td> 3.18</td><td>(s,</td><td>3H),</td>

<td>2.52 (s,</td><td>3H)</td><td> , 2.</td><td>35 (p,</td><td>J =</td><td>8.i hz, ih:</td><td> ) , 2</td><td> .16 -</td><td> 1.89</td><td>(m,</td><td>6H),</td>

<td> 1.77 - 1.</td><td> 53</td><td>(m,</td><td>4H), 1.</td><td> 49</td><td colspan="2">(s, 3H), 1.45 -</td><td> 1.26</td><td>(m,</td><td>5H),</td><td> 1.09</td>

(s, 9H), 0.93 (dd, J = 11.3, 6.7 Hz, 6H).

C33H54N4O5S caled m / z - 618.38 found [M + H] + = 619.54

193

Example 36

OR

<img file="MX368258B_D0093.tif" />

(36) (S) -methyl 2- (tert-butoxycarbonyl (methyl) amino) -3- (4-cyanophenyl) -3-methylbutanoate

To a mixture of the methyl ester of Example 38 (0.06g,

0.15mmol), tris (dibenzylidenacetone) dipaladium (0) (0.014g, 0.015mmol), l, l'-bis (diphenylphosphine) ferrocene (0.02g, 0.25 equiv), magnesium acetate (0.013g, 0.06mmol), powder of zinc (0.004g, 0.06mmol) and zinc cyanide (0.0264g, 0.225mmmol) in a nitrogen bath, N, N-dimethylformamide / water (0.8 / 0.08 ml) was added. The reaction was sprayed with nitrogen gas, then the vial was sealed and immersed in an oil bath at 105 ° C. The reaction was allowed to stir overnight and allowed to cool to room temperature. HPLCMS analysis indicated a good conversion into the desired product. The reaction was concentrated under reduced pressure, suspended in CH₂C1₂ and the resulting suspension was purified by silica gel chromatography (15-25% EtOAc in hexanes) to give the final compound as a colorless oil (0.036g, 69%). 1 H NMR (400 MHz, Chloroform-d) δ 7.69 - 7.35 (m, 4H), 5.24 (s, 1H), 3.54 (s, 3H), 2.74 (s, 3H), 1.51 (s, 3H), 1.45 194

1.25 (m, 12H).

<img file="MX368258B_D0094.tif" />

(37) (S) -methyl 2- (tert-butoxycarbonyl (methyl) amino) -3- (4 - ((tertbutoxycarbonylamino) methyl) phenyl) -3-methylbutanoate

To a solution of benzonitrile (0.300 g, 0.87 mmol) in methanol / acetic acid (10: 1.9 ml) in a stirring vessel was added palladium black. The flask was charged with hydrogen gas at 60 psi and the stirrer was turned on for 24 h. At that time, the vessel was purged of H₂ under reduced pressure. The reaction was diluted with methanol and the suspension was filtered through a celite pad. The filtrate was concentrated to a slightly yellow oil and redissolved in dichloromethane (5 mL). To the solution, t-butyl dicarbonate (0.524g, 2.0 equiv) and triethylamine (0.846mL, 5 equiv) were added at 0 ° C with stirring. The reaction was allowed to stir for 3 h, at which time HPLC-MS indicated complete consumption of the amine. The reaction was concentrated under reduced pressure and purified by silica gel chromatography (diethyl ether in hexanes, 15-30%) to give the

195 title compound as a colorless oil (0.232g, 60%).

1H NMR (400 MHz, Chloroform-d) δ 7.38 (dd, J = 16.6, 8.0 Hz, 2H), 7.23 (d, J = 7.7 Hz, 2H), 5.27 (s, 1H), 4.31 (s, 2H) , 3.61 (s, 3H), 2.78 (s, 3H), 1.50-1.61 (m, 6H), 1.47 (d, J = 15.2 Hz, 18H).

Example 38

ΌΗ

Br (38)

(S) -3- (4-Bromophenyl) -2- (tert-butoxycarbonyl (methyl) amino) -3-methylbutanoic acid

To a stirred solution of (S) -methyl-3- (4-bromophenyl) -2- (tertbutoxycarbonyl (methyl) amino) -3-methylbutanoate (0.710g, 1.77mmol) in 1.4 dioxane (4 mL) was added water (1 ml) (2 ml) and lithium hydroxide monohydrate (0.367g, 8.9 mmol). The reaction was heated to 50 ° C and monitored by HPLC until its completion. The reaction was cooled to room temperature, acidified to pH 3 with 1M citric acid and concentrated to almost dryness under reduced pressure. The residue was taken up in ~ 20 ml of ethyl acetate, washed with brine, dried over MgSO₄, was filtered and concentrated to give the analytically pure material that was used without further manipulation.

196

1 NMR (400 MHz, Chloroform-d) δ 7.44 (d, J = 8.3 Hz, 2H), 7.33 (d, J = 8.3 Hz, 2H), 5.18 (s, 1H), 2.71 (s, 3H), 1.60 1.42 (m, 15H).

Example 39

OR

<img file="MX368258B_D0095.tif" />

(39)

(S) -3- (4-azidophenyl) -2- (tertbutoxycarbonyl (methyl) amino) -3-methylbutanic acid

Example 38 (0.690g, 1.8 mmol), copper (I) iodide (0.034g, 0.18 mmol), sodium azide (0.350g, 5.4 mmol) was added to an open pressure tube containing a magnetic stir bar. , NI, N2-dimethylethan-l, 2-diamine (0.029mL, 0.27 mmol), sodium ascorbate (0.036g, 0.18 mmol), sodium hydroxide (0.072 g, 1.8 mmol), ethanol (6 mL) and water (1 mi). The suspension was purged with nitrogen gas, the vessel was sealed and immersed in an oil bath at 105 ° C with vigorous stirring. The course of the reaction was monitored by HPLC-MS within 24 hours at which time there was little left of the starting material. The reaction was diluted with ethyl acetate (~ 20 mL) and washed with brine. The aqueous layer was extracted 2x with ~ 20 ml of ethyl acetate. The

197 organic layers were combined, dried over MgSO₄, filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography (20-65% EtOAc (containing 2% v / v AcOH) in hexanes) to give the title compound as a colorless oil (0.475g, 75%).

1H NMR (400 MHz, Chloroform-d) δ 7.44 (d, J = 8.6 Hz, 2H), 6.99 (dd, J = 9.0, 3.4 Hz, 2H), 5.24 (s, 1H), 2.71 (s, 3H) , 1.63 - 1.38 (m, 18H).

Example 40

<img file="MX368258B_D0096.tif" />

Chemical formula: C35H49N5O5S

Exact mass: 651.35 (40) (S, E) -N- (benzylsulfonyl) -4 - ((S) -2 - ((5) -3- (4-cyanophenyl) -3methyl-2- (methylamino) butanamide) -N, 3,3-trimethylbutanamide) 2,5-dimethylhex-2-enamide

The title compound was prepared according to Example 36 and (S, E) -4 - ((S) -2-amino-N, 3,3-trimethylbutanamido) -N (benzylsulfonyl) -2,5-dimethylhex-2 -enamide using General Procedures 3, 4 and 7.

* H NMR (400 MHz, Methanol-d4) δ 7.83 (d, J = 8.2 Hz, 2H), 7.73

198

<td>(d,</td><td colspan="2">J = 8.4 Hz, 2H</td><td> ) _F</td><td> 7.</td><td> 38 (</td><td>d, J</td><td> = 2.6</td><td>Hz, 5H),</td><td> 6.39</td><td>(dd,</td><td>J =</td>

<td> 9.2,</td><td>1.8 Hz,</td><td>1 HOUR) ,</td><td> 5.</td><td> . 04</td><td>(t,</td><td>J =</td><td> 10.1</td><td>Hz, 1H),</td><td> 4.91</td><td>(s,</td><td>1 HOUR) ,</td>

<td> 4.75</td><td>(s, 2H),</td><td> 4.34</td><td> (</td><td>s,</td><td>1 HOUR) ,</td><td> 3.12</td><td>(s,</td><td>3H), 2.54</td><td colspan="2">(s, 3H),</td><td> 2.05</td>

<td> - 1.</td><td>97 (m, 2H</td><td> ), 1.</td><td> 95</td><td>(d</td><td>J</td><td> = 1.5</td><td>Hz,</td><td>3H), 1.52</td><td colspan="2">(s, 3H),</td><td> 1.41</td>

(s, 3H), 1.09 (s, 9H), 0.91 (dd, 11.2, 4.8 Hz, 6H).

C35H49N5O5S caled m / z = 651.35 found [M + H]⁺ = 652.4

Example 41

<img file="MX368258B_D0097.tif" />

(S, E) -4 - ((S) —2 - ((3) -3- (4- (aminomethyl) phenyl) -3-methy1-2 (methylamino) butanamide) -N, 3,3-trimethylbutanamide) -N (benzylsulfonyl) -2,5-dimethylhex-2-enamide

The title compound was prepared according to Example 37 and (S, E) -4 - ((S) -2-amino-N, 3,3-trimethylbutanamide) -N (benzyl sulfonyl) -2,5-dimethylhex-2 -enamide using the

General Procedures 3, 4 and 7.

<td>1 HOUR</td><td>NMR (400</td><td>MHz,</td><td colspan="2">Methanol-d4)</td><td>δ 7.</td><td>63 (t, J =</td><td> = 8.8</td><td>Hz,</td><td>2H)</td><td>F</td><td> 7.54</td>

<td>(d,</td><td>J = 8.3</td><td>Hz,</td><td>2H), 7.</td><td> 49 -</td><td> 7.43</td><td>(m, 3H),</td><td> 7.39</td><td>(m,</td><td>2H)</td><td>F</td><td> 6.39</td>

<td>(d,</td><td>J = 9.4</td><td>Hz,</td><td>1H), 5.</td><td> 05 -</td><td> 4.97</td><td>(m, 1H),</td><td> 4.75</td><td>(s,</td><td>2H)</td><td>F</td><td> 4.35</td>

<td>(s,</td><td>3H), 4.</td><td>16 (s</td><td>, 2H),</td><td> 3.14</td><td>(s,</td><td>3H), 2.54</td><td>(s,</td><td>3H),</td><td> 2.</td><td> 03</td><td>(m,</td>

<td>1 HOUR)</td><td> , 1.95 (</td><td>s, 3H</td><td> ), 1-51</td><td>(s,</td><td>3H),</td><td>1.39 (s,</td><td>3H),</td><td> 1.31</td><td>(s</td><td>t</td><td>3H),</td>

199

1.09 (s, 9H), 0.98-0.81 (m, 6H).

She

Example 42

<img file="MX368258B_D0098.tif" />

Chemical formula: C ^ H ^ NrOgS

Exact mass: 667.35 (42) (S, E) -4 - ((S) -2 - ((S) -3- (4-azidophenyl) -3-meti1-2 (methylamino) butanamide) -N, 3, 3-Trimethylbutanamido) -N (benzylsulfonyl) -2,5-dimethylhex-2-enamide title compound was prepared according to Example 39 and (S, E) -4 - ((S) -2-amino-N, 3, 3-Trimethylbutanamido) -N (benzylsulfonyl) -2,5-dimethylhex-2-enamide using the

General Procedures 4 and 7.

C34H49N7O5S caled m / z = 667.35 amu; found [M + H]

668.4

<img file="MX368258B_D0099.tif" />

Chemical formula:

Exact mass:

C34H51N5O5S

641.36 (43)

200 * '' I ♦ ís. w 1 *&gt; »(S, E) -4 - ((S) —2 - ((S) —3— (4-aminophenyl) -3-methyl-2 (methylamino) butanamide) -N, 3,3- trimethylbutanamide) -N (benzylsulfonyl) -2,5-dimethylhex-2-enamide

To a stirred solution of Example 42 protected with Boc (0.035 g, 0.046mmol) in ethanol (1.6 ml) and water (0.5 ml) was added zinc powder (0.015g, 0.23 mmol) and ammonium chloride (0.025g, 0.46 mmol). After 1 h, HPLC-MS indicated the complete consumption of the starting material. The reaction was quenched with ammonium hydroxide (~ 0.1 ml) and diluted with ethyl acetate (5 ml). The reaction was filtered, the solids were washed with ethyl acetate (5 mL) and the biphasic filtrate was transferred to a separatory funnel. The aqueous phase was washed twice with ethyl acetate (5 ml) and the organic phases were combined, washed with brine, dried over MgSO₄, filtered and concentrated. The reaction product was purified by silica gel chromatography (515% MeOH in CH₂C1₂) to provide the Boc protected intermediate as a colorless glass (0.027g, 66%). The intermediary was checked out in accordance with General Procedure 7 to give the title compound.

C34H51N5O5S caled m / z = 641.36 amu; found [M + H]⁺ = 642.4

201

Example 44

<img file="MX368258B_D0100.tif" />

Chemical formula: C ^ H ^ N ^ S

Exact mass: 618.38 (44) (S, E) -N- (cyclohexylsulfonyl) -2,5-dimethyl-4 - ((S) -N, 3,3-trimethyl-2 - ((S) -3-methyl-2 - (methylamino) -3phenylbutanamide) butanamide) hex-2-enamide

The title compound was prepared according to Example 3 and cyclohexylsulfonamide using General Procedures 2 and 7.

1H NMR (400 MHz, Methanol-d4) δ 7.61 - 7.52 (m, 2H), 7.47 (dd, J = 8.6, 6.9 Hz, 2H), 7.36 (t, J = 7.5 Hz, 1H), 6.61 - 6.50 ( m, 1H), 5.11-499 (m, 1H), 4.94 (s, 1H), 4.28 (s, 1H), 3.59-3.51 (m, 1H), 3.18 (s, 3H), 2.48 (s, 3H) , 2.20 - 2.00 (m, 4H), 1.97 - 1.87 (m, 6H), 1.78 - 1.69 (m, 1H), 1.60 (td, J = 14.2, 10.9 Hz, 2H), 1.48 (s, 3H), 1.44 - 1.23 (m, 6H), 1.09 (s, 9H), 0.93 (dd, J = 13.7, 6.6 Hz, 7H).

C33H54N4O5S caled m / z = 618.38 found [M + H] + = 619.47

202

Example 45

<img file="MX368258B_D0101.tif" />

Chemical formula: C₃₃H₄₉N₅OR₅S

Exact mass: 627.35 (45) (S, E) -2,5-dimethyl-N- (pyridin-3-ylmethylsulfonyl) ~ 4 ~ ((S) -N, 3,3-trimethyl-2 - ((S) -3 -methyl-2- (methylamino) -3phenylbutanamido) butanamido) hex-2-enamide

The title compound was prepared according to Example 3 and pyridin-3-ylmethanesulfonamide using General Procedures 2 and 7.

1 H NMR (400 MHz, Methanol-d4) δ 8.55 (d, J = 1.7 Hz, 1 H), 8.48 (dd, J = 5.0, 1.6 Hz, 1 H), 7.89 (d, J = 8.0 Hz, OH), 7.55 (d, J = 7.6 Hz, 2H), 7.50 - 7.39 (m, 2H ), 7.35 (s, 1H), 6.52 (dd,

J = 9.6, 2.0 Hz, 1H), 5.05 (s, OH), 4.94 (s, 1H), 4.64 (s,

2H), 4.19 (s, 1H), 3.11 (s, 3H), 2.45 (s, 3H), 1.91 (d, J =

1.5 Hz, 3H), 1.48 (s, 3H), 1.39 (s, 3H), 1.07 (s, 8H), 0.89 (dd, J = 15.1, 6.5 Hz, 6H).

C33H54N4O5S caled m / z = 627.35 found [M + H] + = 628.35

203

Example 46

<img file="MX368258B_D0102.tif" />

(46)

4- (N - ((S, E) -2,5-dimethyl-4 - ((S) -N, 3,3-trimethyl-2 - ((S) -3methyl-2- (methylamino) -3 acid -phenylbutanamido) butanamido) hex-2enoyl) sulfamoyl) benzoic

The title compound was prepared according to Example 3 and methyl 4-sulfamoylbenzoate using the Procedures

General 2, 3 and 7.

1 H NMR (400 MHz, Methanol-d4) δ 8.25 - 8.07 (m, 4H), 7.54 (d,

J = 7.8 Hz, 2H), 7.47 (t, J = 7.6 Hz, 2H), 7.37 (t, J = 7.3 Hz, 1H), 6.55 (d, J = 9.3 Hz, 1H), 4.98 (t, J = 9.9 Hz, 1H), 4.92 (s, 1H), 4.36 (s, 1H), 3.16 (s, 3H), 2.51 (s, 3H), 2.06 (q, J = 9.0, 7.7 Hz, 1H), 1.88 ( s, 3H), 1.46 (s, 3H), 1.36 (s, 3H), 1.06 (s, 9H), 0.91 (t, J = 6.0 Hz, 6H)

204

Example 47

<img file="MX368258B_D0103.tif" />

Chemical formula: C35H48F3N5O5S

Exact mass: 723.33

Molecular Weight: 723.85 (47) (S, E) -2,5-dimethyl-N- (3- (2,2,2trifluoroacetamido) phenylsulfonyl) -4 - ((S) -N, 3,3-trimethyl-2 ((S) -3-methyl-2- (methylamino) -3-phenylbutanamido) butanamido) hex-2-enamide

The title compound was prepared according to Example 3 and 2,2,2-trifluoro-N- (3-sulfamoylphenyl) acetamide using General Procedures 2 and 7.

NMR (400 MHz, Methanol-d4) δ 8.49 (p, J = 2.2 Hz, 1H), 7.90

<td colspan="4">(dtd, J = 6.0, 4.8,</td><td colspan="2">2.9 Hz, 2H),</td><td> 7.64</td><td colspan="2">- 7.56 (m, 1H),</td><td> 7.53</td>

<td>(tt,</td><td>J = 5.4,</td><td> 4.3,</td><td> , 1</td><td>.8 Hz, 2H),</td><td> 7.</td><td> .51 -</td><td>7.42 (m, 2H),</td><td> 7 .</td><td> 41 -</td>

<td> 7.28</td><td>(m, 1H),</td><td> 6.56</td><td> -</td><td>6.38 (m, 1 H)</td><td>r</td><td> 4.97</td><td>(s, 1H), 4.90</td><td>(d,</td><td>J =</td>

<td> 3.3</td><td>Hz, 1H),</td><td> 4.35</td><td>(s</td><td>, 1H), 3.16</td><td> (</td><td>d, J</td><td>= 15.5 Hz, 3H</td><td> ) _r</td><td> 2.49</td>

<td>(d,</td><td>J = 14.2</td><td>Hz,</td><td>3H)</td><td> , 2.14 - 2.</td><td> 01</td><td>(m,</td><td>1H), 1.89-1</td><td> .83</td><td>(m,</td>

<td>3H),</td><td> 1.57 - 1</td><td> .28</td><td>(m,</td><td>6H), 1.14</td><td> -</td><td> 0.94</td><td>(m, 9H), 0.95</td><td> -</td><td> 0.85</td>

<td>(m,</td><td>6H).</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>¹³C</td><td>NMR (101</td><td>MHz</td><td></td><td>Methanol-d₄)</td><td>δ</td><td> 172</td><td> .26, 168.81,</td><td> 167</td><td> .10,</td>

205

167.00, 144.95, 141.82, 138.82, 138.47, 135.31, 130.71,

130.38, 128.91, 127.36, 126.65, 126.32, 121.39, 71.20, 66.92,

57.87, 57.78, 42.05, 35.83, 34.15, 32.66, 30.84, 29.79,

26.95, 21.39, 19.84, 19.82, 15.45, 14.03.

¹⁹F NMR (377 MHz, Methanol-d₄) δ -76.96, -77.07.

C₃5H₄g F₃N₅0gS caled m / z = 723.33 amu; found [M + H]⁺ =

724.30, [M + Na]⁺ = 746.30

Example 48 i

<img file="MX368258B_D0104.tif" />

Chemical formula: C₃₃H4₉N₅OR₅S

Exact mass: 627.35 (48) (S, E) -N- (3-aminophenylsulfonyl) -2,5-dimethyl-4 - ((S) -N, 3,3-trimethyl-2 - ((S) -3-methyl -2- (methylamino) -3phenylbutanamido) butanamide) hex-2-enamide

The title compound was prepared according to Example 3 the

2,2,2-trifluoro-N- (3-sulfamoylphenyl) acetamide using General Procedures 2, 3 and 7.

^ςΗ NMR (400 MHz, Methanol-d₄) δ 7.55 (d, J = 7.5 Hz, 2H), 7.51

- 7.45 (m, 2H), 7.43 - 7.20 (m, 4H), 6.97 (d, J = 8.1 Hz,

1H), 6.48 (d, J = 9.4 Hz, 1H), 5.02-489 (m, 2H), 4.36 (s,

1H), 3.17 (s, 3H), 2.50 (s, 3H), 2.14

2.00 (m, 1H), 1.88

206 (d, J = 1.4 Hz, 3H), 1.46 (s, 3H), 1.35 (s, 3H), 1.07 (s

9H), 0.92 (d, J = 6.3 Hz, 3H), 0.90 (s, 3H).

C₃₃H₄₉N₅OR₅S caled, m / z = 627.35 found [M + H]⁺ = 628.36

Example 49

<img file="MX368258B_D0105.tif" />

Chemical formula: C₃₂H47N₅OR₅S

Exact mass: 613.33 (49) (S, E) -2,5-dimethyl-N- (pyridin-3-ylsulfonyl) -4- ((S) -N, 3,3-trimethyl-2 - ((S) -3 -methyl-2- (methylamino) -3phenylbutanamido) butanamido) hex-2-enamide

The title compound was prepared according to Example 3 and pyridin-3-sulfonamide using General Procedures 2, 7.

^XH NMR (400 MHz, Methanol-dJ δ 9.18 (s, 1H), 8.80 (s, 1H),

<td> 8.46</td><td>(dt,</td><td>J = 8.2, 1.8</td><td>Hz,</td><td>1H), 7.65 (dd,</td><td>J =</td><td> 8.1</td><td>, 4.9 Hz,</td>

<td>1 HOUR) ,</td><td> 7.54</td><td>(d, J = 7.3 Hz</td><td>, 2ΗΪ</td><td>), 7.47 (t, J =</td><td> 7.8</td><td>Hz,</td><td>2H), 7.37</td>

<td>(t,</td><td>J = 7</td><td>.3 Hz, 1H), 6.</td><td> 54 (</td><td>d, J = 9.3 Hz,</td><td>1 HOUR) ,</td><td> 5.</td><td> 01 - 4.88</td>

<td>(m,</td><td>2H),</td><td>4.36 (s, 1 H),</td><td> 3.18</td><td>l (s, 3H), 2.51</td><td>(s,</td><td>3H</td><td> ), 2.15 -</td>

<td> 2.01</td><td>(m,</td><td>1H), 1.86 (d,</td><td>J =</td><td>1.4 Hz, 3H), 1.</td><td> .46</td><td>(s,</td><td>3H), 1.33</td>

(s, 3H), 1.07 (s, 9H), 0.92 (d, J = 3.3 Hz, 3H), 0.91 (d, J =

207

3.5 Hz, 3H).

C32H47N5O5S caled, m / z = 613.33 found [M + H]⁺ = 614.23

Example 50

Chemical formula:

Exact mass: 618.29 (50) (S, E) -2,5-dimethyl-N- (thiophen-2-ylsulfonyl) -4 - ((S) -N, 3,3-trimethyl-2 - ((S) -3-methyl-2- (methylamino) -3phenylbutanamido) butanamido) hex-2-enamide

The title compound was prepared according to Example 3 and thiophene-2-sulfonamide using General Procedures 2, and 7.

^XH NMR (400 MHz, Methanol-d₄) δ 7.93 - 7.82 (m, 2H), 7.55 (d, J = 8.3 Hz, 1H), 7.48 (t, J = 7.8 Hz, 2H), 7.37 (t, J = 7.2 Hz, 1H), 7.15 (dd , J = 5.0, 3.8 Hz, 1H), 6.51 (d, J = 9.1 Hz, 1H), 5.02-4.93 (m, 2H), 4.36 (s, 1H), 3.18 (s, 3H), 2.51 (s, 3H), 2.15 - 2.01 (m, 1H), 1.89 (d, J = 1.4 Hz, 3H), 1.46 (s, 3H), 1.34 (s, 3H), 1.08 (s, 9H), 0.93 (d, J = 4.8 Hz, 3H), 0.91 (d, J = 4.7 Hz, 3H).

C3iH₄₆N₄OR₅S₂ caled, m / z = 618.29 found [M + H]⁺ = 619.24

208

Example 51

<img file="MX368258B_D0106.tif" />

Chemical formula:

Ex mass

C33H48N4O5S

628.33 (S, E) -N- (4-hydroxyphenylsulfonyl) -2,5-dimethyl-4 - ((S) -N, 3,3-trimethyl-2 - ((S) -3-methyl-2- (methylamino) -3phenylbutanamide) butanamide) hex-2-enamide

The title compound was prepared according to Example 3 and the

4- (tert-butyldimethylsilyloxy) benzenesulfonamide using the

General Procedures 2, and 7.

<td colspan="2">^:H NMR</td><td colspan="2">(400 MHz, Methanol-d₄</td><td>) δ</td><td colspan="2">7.89 (d, J = 8.8 Hz, 2H),</td><td> 7.55</td>

<td>(d,</td><td>J =</td><td>= 7.0 Hz, 1H</td><td> ), 7.47</td><td>(t,</td><td>J = 7.6 Hz, 2H),</td><td>7.37 (t,</td><td>J -</td>

<td> 7.3</td><td>Hz,</td><td>1H), 6.91</td><td>(d, J =</td><td> 8.9</td><td>Hz, 2H), 6.46 (d</td><td>, J = 9.2</td><td>Hz,</td>

<td>1 HOUR) ,</td><td> 4 .</td><td>97 (d, J =</td><td>10.2 Hz,</td><td>1 HOUR</td><td>.), 4.92 (s, 1H),</td><td>4.33 (s,</td><td>1 HOUR) ,</td>

<td> 3.16</td><td>(s</td><td>, 3H), 2.50</td><td>(s, 3H),</td><td> 2.</td><td>11 - 2.00 (m, 1H),</td><td>1.87 (d,</td><td>J =</td>

<td> 1.4</td><td>Hz,</td><td>3H), 1.46</td><td>(s, 3H),</td><td> 1.</td><td>36 (s, 3H), 1.07</td><td>(s, 9H),</td><td> 0.92</td>

<td>go</td><td>J =</td><td>6.5 Hz, 4H)</td><td> , 0.89 (</td><td>d,</td><td>J = 6.7 Hz, 3H).</td><td></td><td></td>

C33H48N4O6S caled, m / z = 628.33 found [M + H]⁺ = 629.38

209

Example 52

<img file="MX368258B_D0107.tif" />

4- (tritylthiomethyl) benzonitrile

Tritylmercaptane (1.48 g, 5.36 mmol, 1.05 eq) in THF (5 mL) was added dropwise to a stirred suspension of sodium hydride (60% dispersion in mineral oil, 214 mg, 5.36 mmol, 1.05 eq) in THF (5 mi) under N₂ to 0^or C. After 15 min, 4- (bromomethyl) benzonitrile (1OOg, 5.10 mmol, 1.0 eq) in THF (5 mL) was added and the reaction was allowed to reach after 1 h, the CCF indicated the conversion Complete starting material. The reaction was quenched by the addition of saturated ammonium chloride, then some dH2O. The mixture was extracted three times with ether, washed with saturated brine, dried over sodium sulfate, and concentrated to a viscous yellow oil. Purification by flash chromatography gave the title compound (1.76 g, 88%) as a light white powder.

<td>^ΤΗ NMR (400</td><td>MHz, Chloroform-d) δ</td><td>7.52 (d,</td><td>J =</td><td> 8.2</td><td>Hz,</td><td>2H),</td>

<td>7.47 (d, J</td><td>= 7.1 Hz, 6H), 7.33 (t,</td><td>J = 7.5</td><td>Hz,</td><td>6H),</td><td> 7.26</td><td>(t,</td>

<td>J = 7.2 Hz,</td><td>3H), 7.19 (d, J = 8.2</td><td>Hz, 2H),</td><td> 3.40</td><td>(s,</td><td>2H).</td><td>m / z</td>

<td>caled for</td><td>C27H21NS = 391.14. Found</td><td>[M + Na]⁺ =</td><td> 414 .</td><td> 13.</td><td>R_F =</td><td> 0.32</td>

210 (10% EtOAc / Hex).

Example 53

TrtS

<img file="MX368258B_D0108.tif" />

NH₂ (53)

1- (4- (tritylthiomethyl) phenyl) cyclopropanamine

4- (tritylthiomethyl) benzonitrile (1.47g, 3.75 mmol, 1.0 eq) was taken up in 40 ml of THF, under an atmosphere of N₂, then cooled to -78 ° C. To this solution was added Ti (O-iPr)₄(1.21ml, 4.13 mmol, 1.1 eq), then ethylmagnesium bromide (3 M, 2.75 mL, 8.26 mmol, 2.2 eq) was added dropwise over 5 min. The dry ice bath was removed, allowing the solution to reach room temperature. After 45 min at rt, BF₃-Et₂O (0.93 ml, 7.51 mmol, 2.0 eq) was added to the now very dark reaction mixture. After stirring for an additional 2.5 h, the reaction was quenched with 5 ml of 2M HC1, followed by pH adjustment to a strong base with approximately 15 ml of 2M NaOH. Some water was added to the mixture, then extracted three times with 75 ml of EtOAc, washed once with dH₂Or, once with saturated brine, it was dried over sodium sulfate, and concentrated to a clear oil. The material was purified by flash chromatography to provide the title compound.

211 (680 mg, 36%) as a clear oil.

¹H NMR (400 MHz, Chloroform-d) δ 7.49 (d, J = 7.8 Hz, 6H), 7.33 (t, J = 7.7 Hz, 6H), 7.26 (t, J = 7.2 Hz, 3H), 7.20 (d , J = 8.2 Hz, 2H), 7.11 (d, J = 8.2 Hz, 2H), 3.32 (s, 2H), 1.06 (dd, J = 7.9, 5.0 Hz, 2H), 0.95 (dd, J = 7.9, 4.7 Hz, 2H). m / z caled, for C₂₉H₂₇NS = 421.19. Found [M + H]⁺ = 422.19.

Rf = 0.21 (50% EtOAc / Hex).

Example 54

V7 ⁰ h ^CF’ ^H (54)

2,2,2-trifluoro-N- (1- (4- (tritylthiomethyl) phenyl) cyclopropyl) acetamide

To a stirred solution of l— (4— (tritylthiomethyl) phenyl) cyclopropanamine (680 mg, 1.61 mmol, 1.0 eq) in CH₂C1₂ trifluoroacetic anhydride (0.448 ml, 3.22 mmol, 2.0 eq) and triethylamine (0.45 ml, 3.22 mmol, 2.0 eq) were added. After two hours, the CCF and HPLC indicated the complete conversion of the starting material. The reaction was quenched by the addition of 3 ml of NaHCO₃, then some dH was added₂Or, and the mixture was extracted three times with CH₂C1₂. The combined organic extracts were washed with saturated brine, dried over sulfate.

212 sodium, and concentrated to a yellow foam, giving the title compound (715 mg, 86%) with sufficient purity to proceed to the next stage.

NMR (400 MHz, Chloroform-d) δ 7.48 (d, J = 7.7 Hz, 6H), 7.32 (t, J = 7.6 Hz, 6H), 7.25 (t, J = 7.2 Hz, 3H), 7.19 (d, J = 8.2 Hz, 2H), 7.10 (d, J = 8.3 Hz, 2H), 6.83 (s, 1H), 3.31 (s, 2H), 1.40 - 1.24 (m, 4H). m / z caled, for C31H26F3NOS = 517.17. Found [M + Na]⁺ = 540.25. R_F = 0.71 (50% EtOAc / Hex).

Example 55

<img file="MX368258B_D0109.tif" />

(55)

2,2,2-trifluoro-N- (1- (4- (mercaptomethyl) phenyl) cyclopropyl) acetamide

2,2,2-trifluoro-N - (1- (4 - (tritylthiomethyl) phenyl) cyclopropyl) acetamide (715 mg, 1.38 mmol, 1.0 eq) in 5 mL CH2CI2 was treated with 2.5 mL TEA. After 1 min, TIPSH (0.42 mL, 2.1 mmol, 1.5 eq) was added, causing the yellow color to fade. After 30 min, the CCF indicated that the reaction was complete. The mixture was concentrated, then co-evaporated once with CH2CI2 and twice with toluene. The residue was purified by flash chromatography to

213 provide the title compound (261 mg, 69%) as a white solid. NMR (400 MHz, Chloroform-d) δ 7.35 - 7.23 (m, 4H), 6.87 (s, 1H), 3.74 (d, J = 7.6 Hz, 2H), 1.77 (t, J =

7.6 Hz, 1H), 1.36 (s, 4H). R_F = 0.47 (20% EtOAc / Hex).

Example 56

<img file="MX368258B_D0110.tif" />

(56)

2,2,2-trifluoro-N- (1- (4- (sulfamoylmethyl) phenyl) cyclopropyl) acetamide.

To a stirred solution of 2,2,2-trifluoro-N- (1- (4 (mercaptomethyl) phenyl) cyclopropyl) acetamide (220 mg, 0.799 mmol, 1.0 eq) in acetonitrile was added dH₂Or (0.029 mi,

1.6 mmol, 2.0 eq), tetrabutylammonium chloride (110 mg, 0.40 mmol, 0.5 eq), then N-chlorosuccinimide (320 mg, 2.40 mmol, 3.0 eq). After 20 minutes, the presence of starting material was not observed by TLC. After 90 min, NH was added₄OH (0.18 ml, 3.2 mmol, 4.0 eq) concentrated. After 10 minutes, 1 ml of NH4CI was added, and the mixture was extracted three times with EtOAc. The combined organic extracts were washed twice with dH₂Or, once with saturated brine, it was dried over sodium sulfate, and concentrated to a clear oil. The residue is

214 purified by flash chromatography to provide the title compound (192 mg, 74%) as a white solid.

<td>^XH NMR (400</td><td>MHz,</td><td>DMSO-d₆) δ</td><td>10.21 (s,</td><td>1 HOUR) ,</td><td> 7.31</td><td>(d, J =</td><td> 8.2</td>

<td>Hz, 2H), 7.</td><td>16 (d</td><td>, J = 8.3</td><td>Hz, 2H),</td><td> 6.85</td><td colspan="2">(s, 2H), 4.23</td><td>(s,</td>

<td>2H), 1.27</td><td>(dt,</td><td>J = 6.1,</td><td>2.3 Hz,</td><td>4H).</td><td>Rf =</td><td> 0.26</td><td> (50%</td>

EtOAc / Hex).

Example 57

<img file="MX368258B_D0111.tif" />

Chemical formula: Cg ^ H ^ NgOeS

Exact mass: 777.37 (57) (S, E) -2,5-dimethyl-N- (4- (1- (2,2,2-trifluoroacetamido) cyclopropyl) benzyl sulfonyl) -4 - ((S) -N, 3, 3trimethyl-2 - ((S) -3-methyl-2- (methylamino) -3phenylbutanamido) butanamide) hex-2-enamide

The title compound was prepared according to Example 3 and Example 56 using General Procedures 2, and 7.

^XH NMR (400 MHz,. Methanol-d₄) δ 7.56 (d, J = 8.4 Hz, 2H), 7.48 (t, J = 7.7 Hz, 2H), 7.37 (t, J = 7.4 Hz, 1H), 7.32 (d, J =

8.5 Hz, 2H), 7.28 (d, J = 8.5 Hz, 2H), 6.37 (d, J = 9.6 Hz,

1H), 5.07 (t, J = 10.0 Hz, 1H), 4.94 (s, 1H), 4.72 (s, 2H),

215

4.37 (s, 1H), 3.13 (s, 3H), 2.52 (s, 3H), 2.08 - 1.96 (m,

1H), 1.96 (d, J = 1.5 Hz, 3H), 1.49 (s, 3H), 1.40 (s, 3H),

<td> 1.35 - 1</td><td> .27</td><td>(m, 4H), 1.10</td><td>(s, 9H),</td><td> 0.92</td><td>(d,</td><td>J = 7.1</td><td>Hz, 3H)</td>

<td>0.89 (d,</td><td>J =</td><td>6.8 Hz, 3H).</td><td></td><td></td><td></td><td></td><td></td>

<td> 5 ¹³NMR</td><td> (101</td><td>MHz, MeOD)</td><td>δ 170.93,</td><td> 168</td><td> .81,</td><td> 165.64,</td><td> 143.58</td>

<td> 142.24,</td><td> 136.</td><td> 87, 134.19,</td><td> 130.64,</td><td> 129.</td><td> 00,</td><td> 127.63,</td><td> 127.53</td>

125.95, 125.61, 69.90, 57.10, 57.02, 56.39, 40.73, 34.55,

34.25, 32.80, 30.60, 29.33, 28.39, 25.57, 20.11, 18.38,

18.34, 16.21, 16.15, 14.04, 12.85.

C39H54F3N5O6S caled, m / z = 777.37 found [M + H]⁺ = 778.55

Example 58

<img file="MX368258B_D0112.tif" />

Chemical formula: C₃₇H₅₅N₅OR₅S

Exact mass: 631.39 (58) (S, E) -N- (4- (1-aminocyclopropyl) benzylsulfonyl) -2,5-dimethyl-4 ((S) -N, 3,3-trimethyl-2 - (( S) -3-methyl-2- (methylamino) -3phenylbutanamide) butanamide) hex-2-enamide

The title compound was prepared according to Example 3 and Example 56 using General Procedures 2, 3 and 7.

⁴H NMR (400 MHz, Methanol-d₄) δ 7.56 (d, J = 8.7 Hz, 2H), 7.51 (s, 4H), 7.47 (t, J = 7.6 Hz, 2H), 7.37 (t, J = 7.3 Hz, 1H),

216

<td> 6.49</td><td>(d, c</td><td> 7=9</td>

<td>1 HOUR) ,</td><td> 4.81</td><td>(d,</td>

<td> 4.39</td><td>(s,</td><td>1 HOUR) ,</td>

<td>1 HOUR) ,</td><td> 1. 97</td><td>(d,</td>

<td>2H),</td><td> 1.40</td><td>(s,</td>

<td>(d,</td><td>J = 6.</td><td>2 Hz</td>

3H), 0.90

3H), 1.34 ¹³NMR (101 MHz, MeOD)

137.54

125.98

34.51

18.39

137.12

134.38

1H), 5.07 (t

1.5 Hz, 3H), 1.

14.0 Hz, 1H)

3.16 (s, 3H), 2.52

<td>J = 10.0 Hz, 1H),</td><td> 4.94</td><td>(s</td>

<td>4.77 (d, J = 13.8</td><td>Hz,</td><td>1 HOUR)</td>

<td>! (s, 3H), 2.11 -</td><td> 1.99</td><td>(m</td>

<td>49 (s, 8H), 1.45 -</td><td> 1.41</td><td>(m</td>

2H), 1.10 (s, 9H), 0.93

1.26 (m

Hz, 3H).

δ 170.94

169.00, 165.69

143.57

131.43, 129.66, 128.98

69.85, 65.51, 57.68, 57.15, 56.39

32.80, 30.68, 29.42, 28.40, 25.61

14.05, 12.86, 11.80.

C37H55N5O5S caled, m / z = 681.39 found [M + H]

Example 59

<img file="MX368258B_D0113.tif" />

nh₂ (59) —phenylcyclopropanamine

The title compound was prepared as

127.51

Bertus, P.

Szymoniak, J.

J. Org. Chem., 2003

40.72

20.14

36.16

18.42

682.49 described in

68, 7133-7136 from benzonitrile mL, 9.7 mmol) to give 270 mg ^:H NMR (400

MHz, Chloroform-d) δ 7.44

7.28 (m, 4H), 7.27

7.15 (m, 1H), 1.18

2H), 1.07

0.95 (m, 2H). R_F

217

0.28 (5% (5% NH₄OH / MeOH) / CH₂C1₂) .

Example 60

<img file="MX368258B_D0114.tif" />

H n ^ cf₃

O (60)

2,2,2-trifluoro-N- (1-phenylcyclopropyl) acetamide

To a stirred solution of 1-phenylcyclopropanamine (270 mg, 2.03 inmol, 1.0 eq) in dioxane (5 ml), trifluoroacetic anhydride (0.310 ml, 2.23 inmol, 1.1 eq) was added. After 5 min, the CCF indicated the complete conversion of the starting material. The mixture was concentrated, then coevaporated once with CH₂C1₂ and once with toluene to give the title compound (453 mg, 97%) as a white powder flakes.

¹H NMR (400 MHz, Chloroform-d) δ 7.47 - 7.15 (m, 5H), 6.88 (s, 1H), 1.65 (s, 4H). m / z caled, for CnH10F3NO = 229.07. Found [M + H]⁺ = 230.14. Rf = 0.82 (5% (5%

NH₄OH / MeOH) / CH₂C1₂)

Example 61

<img file="MX368258B_D0115.tif" />

CF₃

<img file="MX368258B_D0116.tif" />

O (61)

2,2,2-trifluoro-N- (1- (4-sulfamoylphenyl) cyclopropyl) acetamide

218

A stirring chlorosulfonic acid (0.78 ml, 11.8 mmol, 6.0 eq) at 0^or C, solid 2,2,2-trifluoro-N- (1-phenylcyclopropyl) acetamide (450 mg, 1.96 mmol, 1.0 eq) was added portionwise, keeping the temperature low. After complete addition, the mixture was heated to 50 ° C. After 10 minutes, the evolution of gas ceased, and the reaction was allowed to cool. The mixture was slowly added to a glass of ice, being aware of the splashes. The solid that was left on the ice was filtered off. This solid was dried in vacuo and then collected in THF (4 mL). NH₄Concentrated OH (0.44 ml, 7.85 mmol, 4.0 eq) was added, turning the solution green-black. After 2 min, the CCF indicated the complete consumption of the sulfonyl chloride intermediate. 2M HC1 was added until the color faded,

<td>then the</td><td>mixture</td><td>it was extracted</td><td>three times</td><td>with</td><td>EtOAc, it</td>

<td>washed once with</td><td>NaHCO₃</td><td>saturated,</td><td>one time</td><td>with</td><td>brine</td>

<td>saturated, dried</td><td>on</td><td>sulfate</td><td>sodium, and</td><td>I know</td><td>concentrated</td>

<td>until you get a</td><td>solid</td><td>scaly.</td><td colspan="2">The material</td><td>gross it</td>

purified by flash chromatography to give the title compound (235 mg, 39%) as a white solid.

^XH NMR (400 MHz, DMSO-d₆) δ 10.28 (s, 1H), 7.76 (d, J = 8.5

Hz, 2H), 7.32 (d, J = 8.1 Hz, 2H), 7.31 (s, 2H), 1.42 - 1.35 (m, 2H), 1.35 - 1.27 (m, 2H). m / z caled, for C11H11F3N2O3S =

219

308.04. Found [M + H]⁺ = 309.07. R_F = 0.27 (50% EtOAc / Hex).

Example 62

<img file="MX368258B_D0117.tif" />

Chemical formula: C₃₈H₅₂F3N₅OR_fiS

Exact mass: 763.36 (62) (S, E) -2,5-dimethyl-N- (4- (1- (2,2,2trifluoroacetamido) cyclopropyl) phenylsulfonyl) -4- ((S) -N, 3, 3trimethyl-2 - ((S) -3-methyl-2- (methylamino) -3phenylbutanamido) butanamide) hex-2-enamide

The title compound was prepared according to Example 3 and Example 61 using General Procedures 2 and 7.

NMR (400 MHz, Methanol-d₄) δ 8.00 (d, J = 8.6 Hz, 2H), 7.55 (d, J = 7.6 Hz, 2H), 7.48 (t, J = 7.7 Hz, 2H), 7.48 - 7.33 (m, 4H), 6.47 (dd , J = 9.4, 1.6 Hz, 1H), 5.00 (t, J = 10.0

Hz, 1H), 4.92 (s, 1H), 4.35 (s, 1H), 3.15 (s, 3H), 2.51 (s,

3H), 2.11 - 2.00 (m, 1H), 1.86 (d, J = 1.4 Hz, 3H), 1.47 (d, J = 6.2 Hz, 3H), 1.45 (s, 2H), 1.43 (s, 2H), 1.38 (s, 3H),

1.06 (s, 9H), 0.91 (d, J = 6.1 Hz, 3H), 0.89 (d, J = 6.2 Hz,

3H).

C37H₅oF3N₅0₆S caled, m / z = 763.36 found [M + H]⁺ = 764.45

Example 63

220

<img file="MX368258B_D0118.tif" />

Chemical formula: C₃₆H₅₃N₅OR₅S

Exact mass: 667.38 (63) (S, E) -N- (4- (1-aminocyclopropyl) phenylsulfonyl) -2,5-dimethyl-4 ((S) -N, 3,3-trimethyl-2 - (( S) -3-methyl-2- (methylamino) -3phenylbutanamide) butanamide) hex-2-enamide

The title compound was prepared according to Example 3 and the

2,2,2-trifluoro-N- (1- (4-sulfamoylphenyl) cyclopropyl) acetamide using General Procedures 2, 3 and 7.

^ΣΗ NMR (400 MHz, Methanol-d₄) δ 8.13 (d, J = 8.5 Hz, 2H), 7.66 (d, J = 8.6 Hz, 2H), 7.55 (d, J = 7.2 Hz, 2H), 7.47 (t, J =

7.6 Hz, 2H), 7.37 (t, J = 1.2 Hz, 1H), 6.50 (dd, J = 9.4, 1.7 Hz, 1H), 5.02 (t, J = 10.0 Hz, 1H), 4.93 (d, J = 4.9 Hz, 1H), 4.38 (s, 1H), 3.16 (s, 3H), 2.51 (s, 3H), 2.12 - 1.99 (m, 1H), 1.84 (d, J = 1.4 Hz, 3H), 1.51 - 1.46 (m, 5H), 1.46 1.42 (m, 2H), 1.38 (s, 3H), 1.07 (s, 9H), 0.91 (dd, J = 6.7,

1.7 Hz, 6H).

C36H53N5O5S caled, m / z = 667.38 found [M + H]

668.40

221

<td></td><td>Example 64 θΥι koñ i Μ HH V _ZNH ⁿ 0 X ⁿ (64)</td>

<td>S, E) -2</td><td>, 5-dimethyl-N- (2-methylbenzylsulfonyl) -4 - ((S) -N, 3,3-trimethyl-2 - ((S) -3-methyl-2- (methylamino) -3- phenylbutanamide) butanamide) hex-2-enamide</td>

The title compound was prepared according to Example 3 and 2-methylbenzylsulfonamide using General Procedures 2 and 7.

^XH NMR (400 MHz, Methanol-d₄) δ 7.61 - 7.52 (m, 2H), 7.48 (t, J

<td>= 7.6 Hz,</td><td>2H), 7.37 (t, J = 7.3 Hz, 1H), 7.30 - 7.23 (m, 3H),</td>

7.22 - 7.14 (m, 1H), 6.48 (dd, J = 9.3, 1.7 Hz, 1H), 5.08 (t,

<td>J = 10.0</td><td>Hz, 1H), 4.94 (s, 1H), 4.81 (s, 2H), 4.34 (s, 1H),</td>

<td>3.15 (s,</td><td>3H), 2.51 (s, 3H), 2.48 (s, 3H), 2.08-200 (m,</td>

<td>1H), 1.98</td><td>(d, J = 1.1 Hz, 3H), 1.49 (s, 3H), 1.40 (s, 3H),</td>

1.10 (s, 9H), 0.93 (d, J = 6.6 Hz, 3H), 0.91 (d, J = 6.6 Hz,

3H).

C35H52N₄0sS caled, m / z = 640.37 found [M + H]⁺ = 641.41

222

Example 65

<img file="MX368258B_D0119.tif" />

Chemical formula: C ^ H ^ NgO / S

Exact mass: 671.34 (65) (S, E) -2,5-dimethyl-N- (4-nitrobenzylsulfonyl) -4 - ((S) -N, 3,3trimethyl-2 - ((S) -3-methyl -2- (methylamino) -3phenylbutanamido) butanamide) hex-2-enamide

The title compound was prepared according to Example 3 and the

4-nitrobenzylsulfonamide using the Procedures

General 2 and 7.

'-H NMR (400 MHz, Methanol-d₄) δ (d, J = 8.7 Hz, 2H), 7.52 (d,

7.7 Hz, 2H), 7.31 (t, J = 7.3

8.18 (d, J = 8.7 Hz, 2H), 7.64

J = 7.5 Hz, 2H), 7.42 (t, J Hz, 1H), 6.55 (d, J = 9.4 Hz,

<td>1 HOUR) ,</td><td>5.04 (t,</td><td>J = 10.0 Hz, 1H),</td><td>4.92 (s,</td><td>1 HOUR) ,</td><td>4.63 (s, 2H),</td>

<td> 3.08</td><td>(s, 3H),</td><td>2.32 (s, 3H), 1.95</td><td>(dt, J =</td><td> 11.4,</td><td>6.6 Hz, 4H),</td>

<td> 1.89</td><td>(d, J =</td><td>1.4 Hz, 3H), 1.46</td><td>(s, 3H),</td><td> 1.38</td><td>(s, 3H), 1.05</td>

<td>(s,</td><td>9H), 0.89</td><td>(d, J = 6.5 Hz, 3H)</td><td>, 0.85 (d,</td><td>, J =</td><td>6.5 Hz, 3H).</td>

C₃₄H₄9N₅O7S caled, m / z = 671.34 found [M + H]⁺ = 672.36

223

Example 66

<img file="MX368258B_D0120.tif" />

Chemical formula: C₃₄H₄₉CIN₄OR₅S

Exact mass: 660.31 (66) (S, E) -N- (4-chlorobenzylsulfonyl) -2,5-dimethyl-4 - ((S) -N, 3,3-trimethyl-2 - ((S) -3-methyl -2- (methylamino) -3phenylbutanamido) butanamide) hex-2-enamide

The title compound was prepared according to Example 3 and the

4-Chlorobenzyl sulfonamide using the Procedures

General 2 and 7.

<td></td><td>NMR</td><td>(400 MHz, Methanol-d₄)</td><td>δ 7</td><td> .56</td><td>(d, J =</td><td>7.9 Hz, 2H),</td><td> 7.48</td>

<td>(t,</td><td>J =</td><td>= 7.6 Hz, 2H), 7.44</td><td> - 7.</td><td> 34 (</td><td>m, 5H),</td><td>6.39 (d, J =</td><td> 9.5</td>

<td>Hz,</td><td>1 HOUR)</td><td>, 5.06 (t, J = 10.0</td><td>Hz,</td><td>1 HOUR)</td><td> , 4.94</td><td>(s, 1H), 4.75</td><td>(s,</td>

<td>2H)</td><td> , 4.</td><td>35 (s, 1H), 3.13 (s,</td><td>, 3H</td><td> ), 2</td><td>.51 (s,</td><td>3H), 2.06 -</td><td> 1.95</td>

<td>(m,</td><td>1 HOUR)</td><td>, 1.95 (d, J = 1.4</td><td>Hz,</td><td>3H)</td><td> , 1-49 (</td><td>) s, 3H), 1.39</td><td>(s,</td>

<td>3H)</td><td>i.</td><td>09 (s, 9H), 0.91 (d,</td><td>J =</td><td> 6.1</td><td>Hz, 3H),</td><td>0.89 (d, J =</td><td> 5.9</td>

Hz, 3H).

C34H49CIN4O5S caled, m / z = 660.31 found [M + H]

661.32

224

Example 67

<img file="MX368258B_D0121.tif" />

Chemical formula:

Exact mass:

C35H52N4O5S

640.37 (67) (S, E) -2,5-dimethyl-N- (phenethylsulfonyl) -4 - ((S) -N, 3,3-trimethyl-

2 - ((S) -3-methyl-2- (methylamino) -3phenylbutanamido) butanamido) hex-2-enamide

The title compound was prepared according to Example 3 and homobenzyl sulfonamide using General Procedures 2 and 7.

^έΗ NMR (400 MHz, Methanol-d₄) δ 7.56 (d, J = 7.6 Hz, 2H), 7.48 (t, J = 7.5 Hz, 2H), 7.38 (t, J = 7.4 Hz, 1H), 7.34 - 7.28

<td>(m, 2H), 7.28-7.20</td><td>(m, 3H), 6.47 (dd, J = 9.2, 1.7 Hz, 1H),</td>

<td>5.03 (t, J = 10.0 Hz,</td><td>, 1H), 4.94 (s, 1H), 4.36 (d, J = 2.3 Hz,</td>

2H), 3.78 (td, J = 7.5, 4.1 Hz, 2H), 3.17 (s, 3H), 3.12 (t, J

<td>= 7.8 Hz, 2H), 2.51</td><td>(s, 3H), 2.14 - 2.01 (m, 1H), 1.89 (d, J</td>

<td>= 1.4 Hz, 3H), 1.49</td><td>(s, 3H), 1.39 (s, 3H), 1.09 (s, 9H), 0.94</td>

<td>(d, J = 6.6 Hz, 3H),</td><td>0.91 (d, J = 6.6 Hz, 3H).</td>

<td>C35H₅₂N₄OR₅S caled, m / z</td><td>= 640.37 found [M + H]⁺ = 641.36</td>

225

Example 68

<img file="MX368258B_D0122.tif" />

Chemical formula: C ^ H ^ BrhUOsS

Exact mass: 704.26 (68) (S, E) -N- (4-bromobenzylsulfonyl) -2,5-dimethyl-4 - ((S) -N, 3,3-trimethyl-2 - ((S) -3-methyl -2- (methylamino) -3phenylbutanamido) butanamide) hex-2-enamide

The title compound was prepared according to Example 3 and 4-bromobenzyl sulfonamide using General Procedures 2 and 7.

^XH NMR (400 MHz, Methanol-d₄) δ 7.60 - 7.51 (m, 4H), 7.48 (t, J = 7.7 Hz, 2H), 7.39 (s, 1H), 7.31 (d, J = 8.3 Hz, 2H), 6.38 (d, J = 9.3 Hz , 1H), 5.06 (t, J = 10.0 Hz, 1H), 4.93 (s, 1H),

<td>4.74 (s, 2H), 4.36 (s,</td><td>1 HOUR) ,</td><td> 3.13</td><td>(s, 3H),</td><td> 2.52 (</td><td>S,</td><td>3h:</td><td> ), 2.03</td>

<td>- 1.98 (m, 1H), 1.95 (</td><td>d, J =</td><td> = 1.4</td><td>Hz, 3H),</td><td> 1.49 (</td><td>S,</td><td>3h:</td><td> ), 1.39</td>

<td>(s, 3H), 1.09 (s, 9H),</td><td> 0.91</td><td>(d, J</td><td>= 6.1 Hz</td><td>, 3H),</td><td> 0.</td><td> 89</td><td>(d, J =</td>

<td>6.3 Hz, 3H)</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>C₃₄H₄gBrN₄O5S caled, m / z</td><td> = 704</td><td colspan="2">.26 found</td><td>[M + H]⁺</td><td> =</td><td> 705</td><td> .23</td>

226

Example 69

<img file="MX368258B_D0123.tif" />

Chemical formula: C₃₅H₄₉N₅OR₅S Exact mass: gg-, gg (69) (S, E) -N- (4-cyanobenzyl sulfonyl) -2,5-dimethyl-4 - ((S) —N, 3,3— trimethyl-2 - (( S) -3-methyl-2- (methylamino) -3phenylbutanamide) butanamide) hex-2-enamide

The title compound was prepared according to Example 3 and the

4-cyanobenzyl sulfonamide using the Procedures

General 2 and 7.

^ςΗ NMR (400 MHz, Methanol-d₄) δ 7.77 (d, J = 8.3 Hz, 2H), 7.64

- 7.53 (m, 4H), 7.48 (t, J = 7.7 Hz, 2H), 7.38 (t, J = 7.3

<td>Hz,</td><td>1 HOUR) ,</td><td> 6.41</td><td>(dd,</td><td>J =</td><td>9.3, 1.7 Hz, 1H</td><td> ) ,</td><td>5.05 (t,</td><td>J =</td><td> 10.0</td>

<td>Hz,</td><td>1 HOUR) ,</td><td> 4.94</td><td>(s,</td><td>1 HOUR) ,</td><td>4.87 (s, 2H), 4.</td><td> 36</td><td>(s, 1H),</td><td> 3.14</td><td>(s,</td>

<td>3H),</td><td> 2.52</td><td>(s,</td><td>3H),</td><td> 2.06</td><td>- 1.98 (m, 1 H),</td><td> 1.</td><td>95 (d, J =</td><td> = 1.4</td><td>Hz,</td>

<td>3H),</td><td> 1.49</td><td>(s,</td><td>3H),</td><td> 1.39</td><td>(s, 3H), 1.09 (</td><td>s,</td><td>9H), 0.91</td><td>(d,</td><td>⁼</td>

4.0 Hz, 3H), 0.90 (d, J = 4.0 Hz, 3H).

C3₅H₄₉N5O₅S caled, m / z = 651.35 found [M + H]⁺ = 652.38

227

Example 70

<img file="MX368258B_D0124.tif" />

Chemical formula: C₃₄H4₉N₅OR₇S

Exact mass: 671.34 (70) (S, E) -2,5-dimethyl-N- (3-nitrobenzyl sulfonyl) -4- ((S) -N, 3,3-trimethyl-2 - ((S) -3-methyl -2- (methylamino) -3phenylbutanamido) butanamide) hex-2-enamide

The title compound was prepared according to Example 3 and the

3-nitrobenzyl sulfonamide using the Procedures

General 2 and 7.

<td>^XH NMR (400 MHz,</td><td>Methanol-d₄)</td><td>δ</td><td> 8.29</td><td>(d,</td><td>J =</td><td> 8.0</td><td>Hz, 1H),</td><td> 8.26</td>

<td>(s, 1H), 7.83 (d</td><td>, u = 7.8 Hz</td><td>F</td><td>1 HOUR) ,</td><td> 7.67</td><td>(t,</td><td>J =</td><td>8.0 Hz,</td><td>1 HOUR) ,</td>

<td>7.56 (d, J = 7.2</td><td>Hz, 2H), 7.</td><td> 48</td><td>(t,</td><td>J =</td><td> 7.7</td><td>Hz,</td><td>2H), 7.38</td><td>(t,</td>

<td>J = 7.3 Hz, 1H),</td><td>6.43 (dd, J</td><td> =</td><td> 9.4,</td><td> 1.7</td><td>Hz,</td><td>1 HOUR) ,</td><td>5.05 (t,</td><td>J =</td>

<td colspan="2">10.0 Hz,</td><td>1 HOUR) ,</td><td>4.93 (s,</td><td>2H),</td><td>4.93 (s, 1 H),</td><td>4.36 (s, 1 H),</td><td> 3.</td><td> 13</td>

<td>(s,</td><td>3H),</td><td> 2.52</td><td>(s, 3H),</td><td> 2.08</td><td>- 1.98 (m, 1H)</td><td>, 1.96 (d, J</td><td> = 1</td><td> .4</td>

<td>Hz,</td><td>3H),</td><td> 1.48</td><td>(s, 3H),</td><td> 1.39</td><td>(s, 3H), 1.07 1</td><td>¡S, 9H), 0.89</td><td>(d,</td><td>J</td>

<td> = 6</td><td>.6 Hz,</td><td>3H),</td><td>0.88 (d,</td><td>J =</td><td>6.6 Hz, 3H).</td><td></td><td></td><td></td>

C34H49N5O7S caled, m / z = 671.34 found [M + H]⁺ = 672.39

228

Example 71

<img file="MX368258B_D0125.tif" />

Chemical formula: C₃₈H₅₈N₄OR₅S

Exact mass: 682.41 (71) (S, E) -N- (4-tert-butylbenzyl sulfonyl) -2,5-dimethyl-4- ((S) N, 3,3-trimethyl-2 - ((S) - 3-methyl-2- (methylamino) -3phenylbutanamido) butanamido) hex-2-enamide

The title compound was prepared according to Example 3 and the

4-t-butylbenzyl sulfonamide using the Procedures

General 2 and 7.

<td colspan="2">NMR</td><td>(400 MHz, Methanol-d₄)</td><td colspan="2">δ 7.56 (d,</td><td>J =</td><td>7.6 Hz, 2H),</td><td> 7.48</td>

<td>(t,</td><td>J =</td><td>: 7.7 Hz, 2H), 7.43</td><td>(d, J</td><td> = 8.2</td><td>Hz,</td><td>2H), 7.38 (t,</td><td>J =</td>

<td> 7.3</td><td>Hz,</td><td>1H), 7.30 (d, J = 8</td><td>.2 Hz</td><td>, 2H),</td><td> 6.39</td><td>(dd, J = 9.4,</td><td> 1.6</td>

<td>Hz,</td><td>1 HOUR)</td><td>, 5.07 (t, J = 10.0</td><td>Hz,</td><td>1H), 4</td><td> . 93</td><td>(s, 1H), 4.72</td><td>(s,</td>

<td>2H),</td><td> 4.</td><td>37 (s, 1H), 3.13 (s</td><td>, 3H)</td><td> , 2.52</td><td>(s,</td><td>3H), 2.06 -</td><td> 1.98</td>

<td>(m,</td><td>1 HOUR)</td><td>, 1.96 (d, J = 1.4</td><td>Hz,</td><td>3H), 1</td><td> .49</td><td>(s, 3H), 1.39</td><td>(s,</td>

<td>3H),</td><td> 1.</td><td>33 (s, 9H), 1.10 (s</td><td>, 9H)</td><td> , 0.92</td><td>(d,</td><td>J = 6.6 Hz,</td><td>3H),</td>

<td> 0.89</td><td>(d</td><td>, J = 6.5 Hz, 3H).</td><td></td><td></td><td></td><td></td><td></td>

C₃₈H₅₈N₄OR₅S caled, m / z = 682.41 found [M + H] ⁺

683.47

229

Example 72

<img file="MX368258B_D0126.tif" />

Chemical formula: C ^ gNgOyS

Exact mass: 671.34 (72) (S, E) -2,5-dimethyl-N- (2-nitrobenzyl sulfonyl) -4 - ((S) -N, 3,3trimethyl-2 - ((S) -3-methyl -2- (methylamino) -3phenylbutanamido) butanamide) hex-2-enamide

The title compound was prepared according to Example 3 and the

2-nitrobenzyl sulfonamide using the Procedures

General 2 and 7.

^ΧΗ NMR (400 MHz, Methanol-d₄) δ 8.03 (dd, J = 8.0, 1.4 Hz, 1H),

<td></td><td> 7.72</td><td>(td,</td><td>J =</td><td>7.5, 1.5 Hz,</td><td>1H), 7.65 1</td><td>(td, J = 7.7, 1.6</td><td>Hz,</td>

<td></td><td>1 HOUR) ,</td><td> 7.60</td><td>(dd</td><td>, J = 7.6, 1.</td><td>6 Hz, 1H),</td><td>7.56 (d, J = 7.2</td><td>Hz,</td>

<td> 15</td><td>2H),</td><td> 7.48</td><td>(t,</td><td>J = 7.7 Hz, 2H</td><td>:), 7.38 (t,</td><td>J = 7.3 Hz, 1H),</td><td> 6.43</td>

<td></td><td>(dd,</td><td>J =</td><td> 9.4,</td><td>1.6 Hz, 1H),</td><td>5.31 (d, J</td><td>= 14.2 Hz, 1H),</td><td> 5.26</td>

<td></td><td>(d,</td><td>J = 1</td><td> .5.3</td><td>Hz, 1H), 5.06</td><td>(t, J = 1C</td><td>10 Hz, 1H), 4.94</td><td>(s,</td>

<td></td><td>1 HOUR) ,</td><td> 4.37</td><td>(s,</td><td>1H), 3.15 (s,</td><td>3H), 2.52</td><td>(s, 3H), 2.08 -</td><td> 1.98</td>

<td></td><td>(m,</td><td>1 HOUR) ,</td><td> 1.96</td><td>(d, J = 1.4</td><td>Hz, 3H), 1.</td><td>49 (s, 3H), 1.39</td><td>(s,</td>

<td> 20</td><td>3H),</td><td> 1.10</td><td>(s,</td><td>9H), 0.92 (d,</td><td>J = 6.6 Hz,</td><td>3H), 0.90 (d, J =</td><td> 6.6</td>

Hz, 3H).

C3₄H₄₉N₅O7S caled, m / z = 671.34 found [M + H]⁺ = 672.39

230

Example 73

<img file="MX368258B_D0127.tif" />

Chemical formula: C₃₅H₅₁N₅OR₇S

Exact mass: 685.35 (73) (S, E) -2,5-dimethyl-N- (4-nitrophenylsulfonyl) -4 - ((S) -N, 3, 3-trimethyl-2 - ((S) -3-methyl -2- (methylamino) -3phenylbutanamido) butanamide) hex-2-enamide

The title compound was prepared according to Example 3 and the

4-nitro-homobenzyl sulfonamide using the Procedures

General 2 and 7.

1 H NMR (400 MHz, Methanol-d4) δ 8.19 (d, J = 8.7 Hz, 2H), 7.58

- 7.51 (m, 4H), 7.47 (t, J = 7.6 Hz, 2H), 7.37 (t, J = 7.3

<td>Hz,</td><td>1 HOUR) ,</td><td> 6.47</td><td>(dd, J</td><td> = 9.5,</td><td> 1.7</td><td>Hz,</td><td>1H), 5.00</td><td>(t,</td><td>J = 10.0</td>

<td>Hz,</td><td>1 HOUR) ,</td><td> 4.93</td><td>(s, 1H)</td><td> , 4.36 (</td><td>S,</td><td>1 HOUR) ,</td><td>3.91 (dd,</td><td>J =</td><td> 14.9, 8.5</td>

<td>Hz,</td><td>1 HOUR) ,</td><td> 3.84</td><td>(dd, J</td><td> = 12.9,</td><td> 8.</td><td>5 Hz</td><td colspan="2">, 1H), 3.28 (t</td><td>, J = 7.5</td>

<td>Hz,</td><td>2H),</td><td> 3.16</td><td>(s, 3H</td><td> ), 2.51</td><td>(s,</td><td>3h:</td><td> ), 2.12 -</td><td> 1.98</td><td>(m, 1H),</td>

<td> 1.87</td><td>(d,</td><td>J =</td><td>1.4 Hz,</td><td>3H), 1.</td><td> 48</td><td>(s,</td><td>3H), 1.39</td><td>(s,</td><td>3H), 1.08</td>

<td>(s,</td><td>9H),</td><td> 0.91</td><td>(d, J =</td><td>6.6 Hz,</td><td>3H)</td><td> , 0.</td><td>91 (d, J =</td><td> 6.6</td><td>Hz, 3H).</td>

C35H51N5O7S caled, m / z = 685.35 found [M + H] + = 686.38

231

Example 74

<img file="MX368258B_D0128.tif" />

Mood formula: CgsH ^ CIN ^ / S

Exact mass: 704.30

Weight ^ ¿7¡4 &lt;) ular 705.30

Methyl 4-chloro-3- (N - ((S, E) -2,5-dimethyl-4 - ((S) -N, 3,3-trimethyl2 - ((S) -3-methyl-2- ( methylamino) -3phenylbutanamido) butanamido) hex-2-enoyl) sulfamoyl) benzoate

The title compound was prepared according to Example 3 and methyl 4-chloro-3-sulfamolbenzoate using General Procedures 2 and 7.

^XH NMR (400 MHz, Methanol-d4) δ 8.80 (d, J = 2.1 Hz, 1H), 8.20

<td>(dd,</td><td>J =</td><td> 8.3,</td><td> . 2.1</td><td>Hz, 1H), 7.71 (d, J</td><td> = 8.3</td><td colspan="3">Hz, 1H), 7.59 -</td>

<td> 7.52</td><td>(m,</td><td>2H),</td><td> 7.47</td><td>(t, J = 7.7 Hz, 2H),</td><td> , 7.40</td><td> - 7.32</td><td>(m,</td><td>1 HOUR) ,</td>

<td> 6.63</td><td> - 6.</td><td> 56 (</td><td>m, 1H)</td><td>, 5.02 (t, J = 10.0</td><td>Hz, 1H</td><td> ), 4.37</td><td>(s,</td><td>1 HOUR) ,</td>

<td> 3.98</td><td>(s,</td><td>3H)</td><td> , 3.18</td><td>(s, 3H), 2.51 (s,</td><td>3H),</td><td> 2.13 -</td><td> 2.00</td><td>(m,</td>

<td>1 HOUR) ,</td><td colspan="2">1.86 (d,</td><td>. J =</td><td>1.4 Ez, 3H), 1.47 (</td><td>s, 3H)</td><td> , 1.37</td><td>(s,</td><td>3H),</td>

<td> 1.06</td><td>(s,</td><td>9H),</td><td> 0.96</td><td>- 0.87 (m, 6H).</td><td></td><td></td><td></td><td></td>

<td> 13_C</td><td>NMR</td><td> (101</td><td>MHz,</td><td>Methanol-d₄) δ 170</td><td> .87,</td><td> 165.65,</td><td> 164</td><td> . 87,</td>

<td> 143.</td><td> 61,</td><td> 137.</td><td colspan="2"> 01, 136.04, 134.29, 133</td><td> .23,</td><td> 131.81,</td><td> 129</td><td> . 16,</td>

128.98, 128.88, 127.50, 125.98, 69.81, 65.53, 57.39, 56.35,

56.15, 55.37, 51.86, 40.70, 34.51, 32.77, 30.80, 29.39,

232

28.44, 26.18, 25.56, 20.06, 18.40, 14.06, 12.74.

C35H49CIN4O7S caled m / z

704.30 amu; found [M + H]

705.25, [M + Na]

727.25

Example 75

<img file="MX368258B_D0129.tif" />

N CF₃

H ³

OO? S 'h₂n (75)

2,2,2-trifluoro-N- (4- (sulfamoylmethyl) benzyl) acetamide

The title compound was synthesized from (aminomethyl) phenyl) methanesulfonamide and commercially available TFAA using General Procedure 1.

^XH NMR (400 MHz, Acetone-d₆) δ 9.05 (s, 1H), 7.48-7.40 (m

2H), 7.40 - 7.32 (m, 2H), 6.17 (s, 1H), 4.56 (d, J = 6.1 Hz

2H), 4.35 (s, 2H)

Example 76

<img file="MX368258B_D0130.tif" />

N

Chemical formula: C₃₇H₅₂F₃N₅OgS

Exact mass: 751.36

Molecular Weight 751.90

<img file="MX368258B_D0131.tif" />

cf₃ (76) (S, E) -2,5-dimethyl-N- (4 - ((2,2,2-trifluoroacetamido) methyl) benzyl sulfonyl) -4 - ((S) -N, 3,3-trimethyl- 2 - ((S) -3-methyl-2233 (methylamino) -3-phenylbutanamido) butanamido) hex-2-enamide The title compound was prepared according to Example 3 and Example 75 using General Procedures 2 and 7.

NMR (400 MHz, Methanol-d₄) δ 7.57 - 7.49 (m, 2H), 7.45 (t, J = 7.5 Hz, 2H), 7.33 (p, J = 8.8, 7.9 Hz, 5H), 6.37 (d, J =

9.7 Hz, 1H), 5.09 - 5.00 (m, 1H), 4.69 (s, 2H), 4.44 (s, 2H), 4.30 (s, 1H), 3.10 (s, 3H), 2.45 (d, 17.5 Hz, 3H), 2.02 -

1.87 (m, 4H), 1.46 (s, 3H), 1.37 (s, 3H), 1.07 (s, 9H), 0.95-0.81 (m, 6H).

¹⁹F NMR (377 MHz, Methanol-d₄) δ -76.94, -77.24.

C37H52F3N5O6S caled m / z = 751.36 amu; found [M + H]⁺ = 752.46, [M + Na]⁺ = 774.38

Example 77 (77) (S, E) -N- (4- (aminomethyl) benzyl sulfonyl) -2,5-dimethyl-4- ((S) N, 3,3-trimethyl-2 - ((S) -3 -methyl-2- (methylamino) -3 phenylbutanamido) butanamido) hex-2-enamide

Prepared from Example 3 and Example 75 using General Procedures 2, 3 and 7.

234 ¹H NMR (400 MHz, Methanol-d₄) δ 7.60 - 7.54 (m, 2H), 7.54 7.50 (m, 4H), 7.47 (d, J = 8.1 Hz, 2H), 7.37 (t, J = 7.4 Hz, 1H), 6.49 (dd, J = 9.5 , 1.5 Hz, 1H), 5.07 (t, J = 10.0 Hz,

1H), 4.94 (s, 1H), 4.83 (d, J = 14.3 Hz, 1H), 4.79 (d, J =

13.9 Hz, 1H), 4.38 (s, 1H), 4.16 (s, 2H), 3.16 (s, 3H), 2.52 (s, 3H), 2.10 - 2.00 (m, 1H), 1.97 (d, J = 1.4 Hz, 3H), 1.49 (s, 3H), 1.40 (s, 3H), 1.10 (s, 9H), 0.93 (d, J = 6.9 Hz,

3H), 0.91 (d, J = 7.0 Hz, 3H).

C35H53N5O5S caled, m / z = 655.4; found [M + H]⁺ = 656.3, [M + 2H] ²⁺ = 328.8.

Example 78

<img file="MX368258B_D0132.tif" />

s'^NH2

OR₂ (78)

2,2,2-trifluoro-N- (4- (sulfamoylmethyl) phenyl) acetamide

The title compound was synthesized from commercially available (4aminophenyl) methanesulfonamide and TFAA using General Procedure 1.

^XH NMR (400 MHz, DMSO-d₆) δ 11.31 (s, 1H), 7.79 - 7.51 (m,

2H), 7.51 - 7.23 (m, 2H), 6.85 (s, 2H), 4.27 (s, 2H).

235

Example 79

<img file="MX368258B_D0133.tif" />

Chemical formula: C₃Hey₅₀F₃N₅OR₆S

Exact mass: 737.34

Molecular Weight 737.87 (79) (S, E) -2,5-dimethyl-N- (4- (2,2,2trifluoroacetamido) benzylsulfonyl) -4 - ((S) -N, 3,3-trimethyl-2 ( (S) -3-methyl-2- (methylamino) -3-phenylbutanamido) butanamido) hex-2-enamide

The title compound was prepared according to Example 3 and Example 78 using General Procedures 2 and 7.

^XH NMR (400 MHz, Methanol-d4) δ 7.68 (d, J = 8.6 Hz, 2H), 7.54 (d, J = 7.1 Hz, 2H), 7.45 (t, J = 7.6 Hz, 2H), 7.37 (dd , J 10.6, 5.0 Hz, 3H), 6.34 (d, J = 9.4 Hz, 1H), 5.04 (t, J = 10.1 Hz, 2H), 4.74 (s, 2H), 4.35 (s, 1H), 3.10 ( s, 3H), 2.49 (s, 3H), 2.02 - 1.94 (m, 1H), 1.93 (d, J = 1.4 Hz, 3H), 1.46 (s, 3H), 1.37 (s, 3H), 1.06 (s , 9H), 0.88 (d, J = 6.3 Hz, 3H), 0.86 (s, 3H).

¹⁹F NMR (377 MHz, Methanol-d4) δ -76.97, -77.05.

C36H50F3N5O6S caled m / z = 737.34 amu; found [M + H]⁺ =

738.38, [M + Na]

760.35

236

Example 80

OR

<img file="MX368258B_D0134.tif" />

NH

N

<img file="MX368258B_D0135.tif" />

nh₂ (80) (S, E) -N- (4-aminobenzyl sulfonyl) -2,5-dimethyl-4 - ((S) -N, 3,3 trimethyl-2 - ((S) -3-methyl-2- (methylamino) -3 phenylbutanamide) butanamide) hex-2-enamide

The title compound was prepared according to Example 3 and Example 78 using General Procedures 2, 3 and 7.

NMR (400 MHz, Methanol-d₄) δ 7.56 (d, J = 7.6 Hz, 2H), 7.48 (t, J = 7.7 Hz, 2H), 7.37 (t, J = 7.3 Hz, 1H), 7.20 (d, J = 8.5 Hz, 2H), 6.87 (d, J = 8.5 Hz, 2H), 6.39 (d, J = 9.4 Hz, 1H), 5.07 (t, 7 = 10.0 Hz, 1H), 4.95 (s, 1H), 4.64 (s, 2H) ,

4.38 (s, 1H), 3.14 (s, 3H), 2.52 (s, 3H), 2.07 - 1.98 (m,

1H), 1.96 (d, J = 1.4 Hz, 3H), 1.49 (s, 3H), 1.39 (s, 3H),

1.10 (s, 9H), 0.92 (d, J = 6.7 Hz, 3H), 0.90 (d, 6.4 Hz,

3H).

C34H51N5O5S caled, m / z = 641.4; found [M + H]⁺ = 642.3.

Example 81 (81)

237

4- (azidomethyl) benzenesulfonamide

To a stirred solution of 4- (bromomethyl) benzenesulfonamide (0.50 g) in N, N-dimethylformamide (1 ml) was added sodium azide (0.20 g). The suspension was heated at 50 ° C for 3 hours at which point the solvent was removed under reduced pressure. The residue was partitioned between ethyl acetate and water. The organic phase was washed with brine, dried over magnesium sulfate, filtered and concentrated to dryness to give the title compound as a syrup which solidified on standing.

^XH NMR (400 MHz, Chloroform-d) δ 8.06-7.91 (m, 2H), 7.58 7.44 (m, 2H), 4.96 (s, 2H), 4.48 (s, 2H).

Example 82

<img file="MX368258B_D0136.tif" />

4- (aminomethyl) benzenesulfonamide

To a solution of 4- (azidomethyl) benzenesulfonamide (0.354g) in methanol (10 ml) in a round bottom flask equipped with a magnetic stirrer was added 10% Pd / C (~ 0.05 g). The flask was purged of gases under reduced pressure and charged with hydrogen. This evacuation and purge was repeated three times in

238 whose point the suspension was left under stirring overnight. After 16 h, the analysis by

CCF indicated the complete consumption of the starting material. The reaction was diluted with methanol (40 ml), celite was added and the mixture was filtered through a resulting solution, it was suggested that the material be used for use.^XH NMR (400 MHz

2H), 3.76 (d

The sintered glass funnel.

concentrated to dryness. The^XH was pure enough later without purification.

DMSO-d₆) δ 7.77 (m, 2H), 7.53 (m, 2H)

J = 11.9 Hz, 2H).

Example 83 h₂n

OR

<img file="MX368258B_D0137.tif" />

H n ^ cf₃

The

NMR is

5.76 (83)

2,2,2-Trifluoro-N- (4-sulfamoylbenzyl) acetamide title compound was synthesized by reaction of

4 (aminomethyl) benzenesulfonamide with TFAA according to

General Procedure 1, with a spectrum of ^XH NMR that was complicated by rotamers.

^XH NMR (400 MHz, DMSO-d₆) δ 7.91 - 7.75 (m

2H), 7.55 - 7.31 (m, 4H), 4.72 (m, 2H), 4.47 (d, J = 6.0 Hz, 1H), 3.18 (s,

239

2Η).

Example 84

<img file="MX368258B_D0138.tif" />

OR

Chemical Formula: CagHgoFaNgOeS

Exact Mass: 737.34

Molecular Weight: 737.87 (84) (S, E) -2,5-dimethyl-N- (4 - ((2,2,2-trifluoroacetamido) methyl) phenylsulfonyl) -4 - ((S) -N, 3, 3-Trimethyl-2 - ((S) -3-methyl-2 (methylamino) -3-phenylbutanamido) butanamido) hex-2-enamide

The title compound was prepared according to Example 3 and Example 83 using General Procedures 2 and 7.

NMR (400 MHz, Methanol-d₄) δ 8.02 (d, J = 8.5 Hz, 2H), 7.58

- 7.42 (m, 7H), 7.35 (t, J = 7.3 Hz, 1H), 6.46 (d, J = 8.5

Hz, 1H), 4.97 (d, J = 10.4 Hz, 1H), 4.54 (s, 2H), 4.33 (s,

1H), 3.14 (s, 3H), 2.48 (s, 3H), 2.11 - 1.97 (m, 1H), 1.83 (d, J = 1.4 Hz, 3H), 1.53 (s, 1H), 1.44 (s, 3H ), 1.34 (s,

3H), 1.04 (s, 9H), 0.89 (d, J = 3.9 Hz, 3H), 0.88 (d, J = 4.1 Hz, 3H).

¹⁹F NMR (377 MHz, Methanol-d₄) δ -76.94, -77.26.

240

C36H50F3N5O6S caled m / z = 737.34 amu; found [M + H]⁺

738.39, [M + Na]⁺ = 760.41

Example 85

<img file="MX368258B_D0139.tif" />

Chemical Formula: N₅OR₅S

Exact Mass: 641.36 (85) (S, E) -N- (4- (aminomethyl) phenylsulfonyl) -2,5-dimethyl-4- ((S) -

N, 3,3-trimethyl-2 - ((S) -3-methyl-2- (methylamino) -3phenylbutanamido) butanamido) hex-2-enamide

Prepared from Example 3 and Example 83 using General Procedures 2, 3 and 7.

^XH NMR (400 MHz, Methanol-d₄) δ 8.13 (d, J = 8.3 Hz, 2H), 7.68

<td>(d,</td><td>J = 8.3 Hz, 2H), 7.55 (d, J</td><td colspan="2">= 7.6 Hz,</td><td>2H),</td><td>7.47 (t,</td><td>J =</td>

<td> 7.7</td><td>Hz, 2H), 7.37 (t, J = 7.3 Hz,</td><td>, 1 HOUR),</td><td> 6.51</td><td>(dd</td><td>, J = 9.2,</td><td> 1.8</td>

<td>Hz,</td><td>1H), 5.01 (t, J = 10.0 Hz,</td><td>1 HOUR) ,</td><td> 4.37</td><td>(s,</td><td>1H), 4.24</td><td>(s,</td>

<td>2H),</td><td>3.17 (s, 3H), 2.51 (s, 3H)</td><td colspan="2"> , 2.13 - 1</td><td> . 97</td><td>(m, 1H),</td><td> 1.84</td>

<td>(d,</td><td>J = 1.4 Hz, 3H), 1.47 (s,</td><td>3H),</td><td> 1.37</td><td>(s,</td><td>3H), 1.07</td><td>(Y,</td>

9H), 0.91 (dd, J = 6.7, 2.0 Hz, 7H).

Ο₃₄Η₅₁Ν₅Ο3 caled m / z = 641.36 amu; found [M + H]⁺ = 642.4

241

Example 86

<img file="MX368258B_D0140.tif" />

Chemical Formula: C₃4H₄₉Brn₄OR₅S

Exact Mass: 704.26

Molecular Weight: 705 75 (86) (S, E) -N- (benzylsulfonyl) -4 - ((S) -2 - ((S) -3- (4-bromophenyl) -3methyl-2- (methylamino) butanamide ) -N, 3,3-trimethylbutanamide) -

2,5-dimethylhex-2-enamide

The title compound was prepared according to Example 38 and (S, E) -4 - ((S) -2-amino-N, 3,3-trimethylbutanamido) -N (benzylsulfonyl) -2,5-dimethylhex-2 -enamide using General Procedures 4 and 7.

<td>* H NMR (400</td><td>MHz,</td><td>Methanol-d₄)</td><td>δ 7.</td><td> 62 (</td><td>t</td><td>J</td><td> = 9.2</td><td>Hz, 2H)</td><td>t</td><td> 7.50</td>

<td>- 7.43 (m,</td><td>2H),</td><td>7.38 (d, J</td><td> = 2.2</td><td>Hz,</td><td>5H</td><td> ) ,</td><td> 6.38</td><td>(dd, J</td><td> =</td><td> 9.5,</td>

<td>1.8 Hz, 1H¡</td><td>i, 5. (</td><td>) 5 (t, J =</td><td> 10.0</td><td>Hz,</td><td>1 HOUR)</td><td></td><td> 4.92</td><td>(s, 1H)</td><td>r</td><td> 4.75</td>

<td>(d, J = 2.</td><td>2 Hz,</td><td>2H), 4.30</td><td>(s,</td><td>1 HOUR) ,</td><td> 3.</td><td> 12</td><td>(s,</td><td>3H), 2.</td><td> 53</td><td>(s,</td>

<td>3H), 2.06 -</td><td> 1.97</td><td>(m, 1H), 1</td><td> . 95 (</td><td>d, J</td><td> =</td><td> 1.</td><td>5 Hz,</td><td>3H), 1.</td><td> 47</td><td>(s,</td>

<td>3H), 1.39 (</td><td>s, 3H:</td><td>), 1.09 (s,</td><td>9H),</td><td> 0.94</td><td> —</td><td> 0.</td><td>8 6 (m,</td><td>6H).</td><td></td><td></td>

C₃4H₄9BrN₄OR₅S caled m / z = 704.26 amu; found [M + H]⁺ =

705.29, [M + Na]

727.36

242

Example 87

<img file="MX368258B_D0141.tif" />

<img file="MX368258B_D0142.tif" />

Chemical Formula: C₄₂H₅₆N₄OR₆S

O Exact Mass: gg

Molecular Weight: 744 98 (87) (S, £) -4 - ((S) -2 - ((S) -3- (4'-acetylbiphenyl-4-yl) -3-methyl-2 (methylamino) butanamide ) -N, 3,3-trimethylbutanamido) -N (benzylsulfonyl) -2,5-dimethylhex-2-enamide

The title compound was prepared according to the General Procedure from Example 86 protected with Boc and 4-acetylphenylboronic acid.

^XH NMR (400 MHz, Methanol-d₄) δ 8.15 - 8.08 (m, 2H), 7.86 7.76 (m, 4H), 7.66 (dd, J = 14.7, 8.4 Hz, 2H), 7.38 (d, J =

4.9 Hz, 5H), 6.39 (d, J = 9.3 Hz, 1H), 5.05 (t, J = 10.1 Hz,

1H), 4.94 (s, 1H), 4.75 (d, J = 4.1 Hz, 2H), 4.37 (d, J = 16.1 Hz, 1H), 3.13 (d, J = 3.4 Hz, 3H), 2.67 (s, 3H), 2.53 (d, J = 11.6 Hz, 3H), 2.01 (s, 1H), 1.96 (d, J = 1.5 Hz, 3H),

1.54 (d, J = 3.7 Hz, 3H), 1.44 (s, 3H), 1.09 (d, J = 2.7 Hz,

9H), 0.96-0.83 (m, 6H).

C₄2H₅₆N₄O6S caled m / z - 744.39 amu; found [M + H]

745.42

243 [M + Na]⁺ = 767.36

Example 88

<img file="MX368258B_D0143.tif" />

Exact Mass: 732.39

Molecular Weight: 732.97 (88) (S, E) -N- (benzylsulfonyl) -4 - ((S) -2 - ((S) -3- (4'-methoxybiphenyl-4il) -3-methyl-2 - (methylamino) butanamide) -N, 3, 3-trimethylbutanamide) -2,5-dimethylhex-2-enamide

The title compound was prepared according to General Procedure 8 from Example 86 protected with Boc and 4-methoxyphenylboronic acid.

^]H NMR (400 MHz, Methanol-d₄) δ 7.74 - 7.53 (m, 6H), 7.38 (d, J

<td>= 4.7 Hz,</td><td>5H),</td><td colspan="2"> 7.08 - 6.</td><td>99 (m, 2H), 6.43-</td><td> 6.35</td><td>(m, 1H), 5.06</td>

<td>(s, 1H),</td><td> 4.94</td><td colspan="2">(s, 1H),</td><td>4.75 (d, J = 4.1</td><td>Hz,</td><td>2H), 4.38 (s,</td>

<td>1H), 3.86</td><td>(s,</td><td>3H),</td><td> 3.13</td><td>(s, 3H), 2.54 (s,</td><td>3H),</td><td>1.99 (d, J =</td>

<td>11.0 Hz,</td><td>1 HOUR) ,</td><td> 1.96</td><td>(d,</td><td>J = 1.5 Hz, 3H),</td><td> 1.51</td><td>(s, 3H), 1.43</td>

<td>(s, 3H),</td><td> 1.09</td><td colspan="2">(s, 9H), 0</td><td>.96 - 0.85 (m, J =</td><td> 6.0,</td><td>5.1 Hz, 6H).</td>

C₄iH₅6N4O₆S caled m / z = 732.39 amu; found [M + H]⁺ = 733.41, [M + Na]⁺ = 755.40

244

Example 89

<img file="MX368258B_D0144.tif" />

Exact Mass: 702.38

Molecular Weight: 702 95 (89) (S, E) -N- (benzlsulfonyl) -4 - ((5) -2- ((5) -3- (biphenyl-4-yl) -3-methyl-2- (methylamino) ) butanamide) -N, 3,3-trimethylbutanamide) -

2,5-dimethylhex-2-enamide

The title compound was prepared according to General Procedure 8 from Example 86 protected with Boc and phenylboronic acid.

NMR (400 MHz, Methanol-d₄) δ 7.86 - 7.51 (m, 6H), 7.48 (t, J

<td>= 7.6 Hz, 2H), 7.43-</td><td>7.33 (m, 6H), 6.39 (d, J = 9.5 Hz, 1H),</td>

<td>5.06 (t, J = 10.1 Hz,</td><td>1H), 4.94 (s, 1H), 4.75 (d, 3.3 Hz,</td>

2H), 4.37 (d, J = 14.4 Hz, 1H), 3.13 (d, J = 3.7 Hz, 3H),

<td>2.55 (d, J = 4.5 Hz,</td><td>3H), 2.06 - 1.97 (m, 1H), 1.96 (d, J =</td>

<td>1.5 Hz, 3H), 1.52 (s,</td><td>3H), 1.44 (d, J = 4.5 Hz, 3H), 1.09 (d,</td>

<td>J = 5.6 Hz, 9H), 0.96</td><td>- 0.83 (m, 6H).</td>

<td>C₄oH5₄N₄0₅S caled m / z =</td><td>702.38 amu; found [M + H]⁺ = 703.40,</td>

245 [M + Na]⁺ = 725.45

Example 90

<img file="MX368258B_D0145.tif" />

Chemical Formula: C43H₅₈N₄O5S

Exact Mass: 742.41

Molecular Weight: 743.01 (90) (S, E) -N- (benzylsulfonyl) -2,5-dimethyl-4 - ((S) -N, 3,3-trimethyl-2 ((S) -3-meti1- 2- (methylamino) -3- (4- (4-methylstyryl) phenyl) butanamido) butanamido) hex-2-enamide

The title compound was prepared according to the Procedure

General 8 from Example 86 protected with Boc and (E) -4-methylstyrylboronic acid.

<td>^:H NMR</td><td>(400 MHz,</td><td colspan="2">Methanol-d₄)</td><td>δ</td><td> 7.</td><td>65 (d, J </td><td> = 8.2</td><td>Hz, 2H),</td><td> 7.54</td>

<td>(d, J =</td><td>8.2 Hz,</td><td>2H),</td><td> 7.47 (</td><td>: d,</td><td>J</td><td>= 7.8 Hz,</td><td>2H),</td><td>7.38 (s,</td><td>5H),</td>

<td> 7.26 -</td><td>7.11 (m,</td><td>4H),</td><td> 6.39</td><td>(d,</td><td>j</td><td>= 9.3 Hz,</td><td>1 HOUR) ,</td><td>5.06 (t,</td><td>J =</td>

<td>10.0 Hz</td><td>:, 1H), 4</td><td> . 97</td><td> - 4.91</td><td>(m,</td><td></td><td>1H), 4.76</td><td>(s,</td><td>2H), 4.36</td><td>(s,</td>

<td>1H), 3.</td><td>12 (d, J</td><td> = 8</td><td>.9 Hz,</td><td>3H)</td><td>r</td><td>2.54 (s,</td><td>3H),</td><td>2.37 (s,</td><td>3H),</td>

<td> 2.05 -</td><td>1.97 (m,</td><td>1 HOUR) ,</td><td> 1.97 -</td><td> 1.</td><td> 93</td><td>(m, 3H),</td><td> 1.49</td><td>(s, 3H),</td><td> 1.41</td>

<td>(s, 3H)</td><td colspan="2">, 1.09 (d, J</td><td> = 3.5</td><td>Hz,</td><td colspan="2">9H), 0.91</td><td>(tq,</td><td>J = 10.8,</td><td> 4.9</td>

Hz, 6H).

246

CnHssN-jOsS caled m / z = 742.41 amu; found [M + H]⁺ = 743.44, [M + Na]⁺ = 765.41

Example 91

<img file="MX368258B_D0146.tif" />

Chemical Formula: C₃₅H₅₂N4O₆S

Exact Mass: 656.36

Molecular Weight: 656.88 (91) (S, E) -N- (benzylsulfonyl) -4 - ((S) -2 - ((S) -3- (4-methoxyphenyl) -3-methyl-2- (methylamino) butanamide) -N, 3,3-trimethylbutanamide) -

2,5-dimethylhex-2-enamide

The title compound was prepared according to the Procedure

General 9 from Example 86 protected with Boc.

Major Diasteroisomer

<td></td><td>NMR (4 00</td><td>MHz,</td><td colspan="2">Methanol-d₄) δ</td><td> 7.44</td><td>(dd,</td><td>J =</td><td> 12.9, 8.6</td><td>Hz,</td>

<td>2H)</td><td> , 7.40 -</td><td> 7.34</td><td>(m, 5H),</td><td> 7.00</td><td>(t, J</td><td> = 8.4</td><td>Hz,</td><td>2H), 6.38</td><td>(d,</td>

<td>J =</td><td>9.2 Hz,</td><td>1 HOUR) ,</td><td>5.05 (t,</td><td>J = 9.</td><td>9 Hz,</td><td>1 HOUR) ,</td><td> 4.93</td><td>(s, 1H),</td><td> 4.75</td>

<td>(d,</td><td>J = 1.8</td><td>Hz,</td><td>2H), 4.</td><td>29 (s,</td><td>1 HOUR) ,</td><td> 3.84</td><td>(s,</td><td>3H), 3.12</td><td>(s,</td>

<td>3H)</td><td>, 2.51 (s</td><td>, 3H)</td><td> , 2.04 -</td><td> - 1.98</td><td colspan="2">(m, 1H), 1.</td><td colspan="2">95 (d, J = 1.4</td><td>Hz,</td>

3H), 1.45 (s, 3H), 1.37 (s, 3H), 1.09 (s, 9H), 0.92

0.86

247 (m, 6H).

Minor Diasteroisomer:

³Η NMR (400 MHz, Methanol-d₄) δ 7.44 (dd, J = 12.9, 8.6 Hz,

2H), 7.40 - 7.34 (m, 5H), 7.00 (t, J = 8.4 Hz, 2H), 6.38 (d,

J = 9.2 Hz, 1H), 4.99 (t, J = 10.1 Hz, 1H), 4.93 (s, 1H),

4.75 (d, J = 1.8 Hz, 2H), 4.26 (s, 1H), 3.82 (s, 3H), 3.11 (s, 3H), 2.47 (s, 3H), 2.04 - 1.98 (m, 1H), 1.92 (d, J = 1.4 Hz, 3H), 1.53 (s, 3H), 1.48 (s, 3H), 0.94 (s, 9H), 0.92 0.86 (m, 6H).

C₃₅H52N4O₆S caled m / z

656.36 amu; found [M + H]⁺

657.35, [M + Na]⁺ = 679.25

Example 92

<img file="MX368258B_D0147.tif" />

Chemical Formula: CsgHgj ^ OgS

Exact Mass: 656.36

Molecular Weight: 656.88 (92) (S, E) -N- (benzylsulfonyl) -4 - ((S) -2 - ((R) -3- (3-methoxyphenyl) -3-methyl-2- (methylamino) butanamide) -N, 3,3-trimethylbutanamide) -

2,5-dimethylhex-2-enamide

The title compound was prepared according to the Procedure

General from (S, E) -N- (benzylsulfonyl) -4 - ((S) -2248 ((S) -3- (3-bromophenyl) -3-methyl-2- (methylamino) butanamide) N , 3,3-trimethylbutanamide) -2,5-dimethylhex-2-enamide protected with Boc. The two diastereomeric products resulted from the diastereomerically impure starting material and were separable by preparative HPLC.

Major Diasteroisomer:

^XH NMR (400 MHz, Methanol-d₄) δ 7.51 - 7.32 (m, 6H), 7.14 7.07 (m, 1H), 7.06 (t, J = 2.2 Hz, 1H), 6.98 - 6.90 (m, 1H), 6.38 (dd, J = 9.6, 1.7 Hz , 1H), 4.99 (t, J = 10.3 Hz, 1H),

4.93 (s, 1H), 4.75 (d, J = 1.8 Hz, 2H), 4.32 (s, 1H), 3.85 (s, 3H), 3.11 (s, 3H), 2.47 (s, 3H), 2.04 - 1.96 (m, 1H),

1.93 (d, J = 1.4 Hz, 3H), 1.54 (s, 3H), 1.47 (s, 3H), 0.96 (s, 9H), 0.89 (dd, J = 6.6, 3.4 Hz, 6H).

Minor Diasteroisomer: Refer to Example 93 (which follows immediately) for data on ^ΧΗ NMR.

C35H52N4O6S caled m / z = 656.36 amu; found [M + H]⁺ = 657.36, [M + Na]⁺ = 679.29

Example 93

MeO

Chemical Formula: C₃₅H₅₂N₄OR₆S Exact Mass: jg

Molecular Weight: mcoo

656.36

656.88 (93)

249 (S, E) -N- (benzylsulfonyl) -4 - ((S) -2 - ((5) -3- (3-methoxyphenyl) -3-methyl-2- (methylamino) butanamide) -N, 3,3- trimethylbutanamide) -

2,5-dimethylhex-2-enamide

The title compound was prepared according to Example 92.

The two diastereomeric products resulted from the diastereomerically impure starting material and were separable by preparative HPLC.

NMR (400 MHz, Methanol-d₄) δ 7.39 (d, J = 5.5 Hz, 6H), 7.11 (dd, J = 4.9, 2.8 Hz, 3H), 6.38 (d, J = 9.4 Hz, 1H), 5.06 (d,

J = 9.5 Hz, 1H), 4.93 (s, 1H), 4.76 (s, 2H), 4.35 (s, 1H),

3.86 (s, 3H), 3.13 (s, 3H), 2.52 (s, 3H), 2.05 - 1.97 (m,

1H), 1.95 (d, J = 1.6 Hz, 3H), 1.46 (s, 3H), 1.38 (s, 3H),

1.09 (s, 9H), 0.90 (t, J = 6.6 Hz, 6H).

C35H₅₂N₄O6S caled m / z = 656.36 amu; found [M + H]⁺ = 657.36, [M + Na]⁺ = 679.32

Example 94

<img file="MX368258B_D0148.tif" />

Chemical formula:

Exact Mass:

Molecular weight:

¢ 36 ^ 54 N4O7S

686.37

686.90 (94) (S, E) -N- (benzylsulfonyl) -4 - ((5) -2 - ((5) -3- (4- (2

250 hydroxyethoxy) phenyl) -3-methyl-2- (methylamino) butanamido) -N, 3,3-trimethylbutanamido) -2,5-dimethylhex-2-enamide

The title compound was prepared as follows: a mixture of Example 86 protected with Boc, Cul (10 mol%), 3,4,7,8-tetramethyl-1, 10-phenanthroline (20 mol%), Cs₂CO₃ (2.5 eq), and ethylene glycol (90 eq) was stirred under N₂ at 130 ° C for 20 h.

The resulting mixture was diluted with H₂Or, it was carefully acidified with 1M citric acid and extracted with CH₂C1₂(5x). The organic extracts were combined, washed with brine (Ix), dried over MgSO₄, filtered, concentrated in vacuo and purified by silica gel column chromatography (eluting with AcOH / EtOAc / hexane mixtures) to provide the cross coupling product that was subsequently deprotected and purified according to General Procedure 7 .

<td>^ςΗ NMR (</td><td> 400</td><td>MHz, Methanol-d₄</td><td>) δ</td><td>7.46 (d, J =</td><td> = 8.8</td><td>Hz, 2H)</td><td></td><td> 7.</td><td> 38</td>

<td>(d, J =</td><td> 2.5</td><td>Hz, 5H), 7.05</td><td>(d,</td><td>J = 8.4 Hz,</td><td>2H),</td><td> 6.38 (</td><td>d,</td><td>J</td><td> =</td>

<td>9.5 Hz,</td><td>1 HOUR)</td><td>, 5.05 (t, J =</td><td> 10</td><td>.1 Hz, 1H),</td><td> 4.93</td><td>(s, 1H)</td><td>t</td><td> 4 .</td><td> 76</td>

<td>(s, 2H),</td><td> 4.:</td><td>&gt; 8 (d, J = 11.0</td><td>Hz,</td><td>1H), 4.13-</td><td> 4.04</td><td>(m, 2H)</td><td></td><td> 3.</td><td> 90</td>

<td>(t, J =</td><td> 4.6</td><td>Hz, 2H), 3.12</td><td>(d,</td><td>J = 6.2 Hz,</td><td>3H),</td><td> 2.50 (</td><td>d,</td><td>J</td><td> =</td>

<td>16.9 Hz,</td><td>3H)</td><td> , 2.05 - 1.97 (</td><td>'m,</td><td>1H), 1.94 (d,</td><td>, J =</td><td colspan="2">11.0 Hz,</td><td>3H</td><td> ) ,</td>

<td> 1.56 - 1</td><td> .34</td><td>(m, 6H), 1.09 (</td><td>s,</td><td>9H), 0.90 (t,</td><td>J =</td><td>6.4 Hz,</td><td colspan="2">6H),</td><td></td>

C₃₆H₅₄N₄OR₇S caled m / z = 686.37 amu; found [M + H]⁺ = 687.42,

251 [M + Na]⁺ = 709.37

Example 95

<img file="MX368258B_D0149.tif" />

Chemical Formula: CggHgg ^ OySg

Exact Mass:; 744.36

Molecular Weight: 745.00 (95)

S-2- (4 - ((S) -4 - ((5) -1 - (((S, E) -2,5-dimethyl-6-oxo-6 (benzyl sulfonamido) hex-4-en-3 -yl) (methyl) amino) -3,3-dimethyl-loxobutan-2-ylamino) -2-methyl-3- (methylamino) -4-oxobutan-2yl) phenoxy) ethyl ethantioate

The title compound was prepared as follows: tributylphosphine (6 eq) was added to a cold solution (0^or C) under stirring di-tert-butyl azodicarboxylate (6 eq) in THF. After 0.5 h, a solution of Example 94 protected with Boc (1 eq) in THF was added, followed by a solution of AcSH (4.5 eq) in THF. The pale yellow mixture was stirred at 0 ° C for 1 h and then at room temperature for 23 h. The resulting mixture was concentrated in vacuo, dissolved in EtOAc and washed successively with 1M HC1 (2x), saturated NH solution.₄C1 (Ix) and brine (lx). The compounds

252 The organics were dried over MgSO4, filtered, concentrated in vacuo and purified by silica gel column chromatography (eluting with AcOH / EtOAc / hexane mixtures) to provide the Boc protected thioacetate product (HPLC / MS - [M + Na] ⁺ = 867.47).

The thioacetate was dissolved in CH2CI2 and treated with TFA. After stirring for 1 h, the reaction mixture was concentrated in vacuo. The yellow / brown residue was dissolved in the minimum amount of CH₂C1₂, cooled to 0 ° C and treated with ether to precipitate the desired aminothioacetate as an off-white solid in 10% yield in two synthetic steps.

NMR (400 MHz, Methanol-d₄) δ 7.46 (d, J = 8.7 Hz, 2H), 7.38

<td>(d,</td><td>J = 2.4 Hz, 5H)</td><td> , 7.03</td><td>(d, J = 8. 6 Hz,</td><td>2H)</td><td>, 6.38 (d,</td><td>J =</td>

<td> 9.5</td><td>Hz, 1H), 5.05</td><td>(t, J =</td><td>= 10.0 Hz, 1H), 4</td><td> . 93</td><td>(s, 1H),</td><td> 4.75</td>

<td>(s,</td><td>2H), 4.27 (d, J</td><td> = 11.4</td><td>Hz, 1H), 4.14 (t,</td><td>J</td><td>= 6.6 Hz,</td><td>2H),</td>

<td> 3.28</td><td>(t, J = 6.6 Hz</td><td>, 2H),</td><td>3.11 (d, J = 6.6</td><td>Hz,</td><td>3H), 2.49</td><td>(d,</td>

<td>J =</td><td>15.5 Hz, 3H), 2.</td><td>.38 (s,</td><td>3H), 2.05 - 1.97</td><td>(m,</td><td>1H), 1.95</td><td>(s,</td>

<td>3H),</td><td>1.45 (s, 3H),</td><td> 1.37 (</td><td>s, 3H), 1.08 (s,</td><td>9H)</td><td> - , 0.96 -</td><td> 0.85</td>

(m, 6H).

C38H56N4O7S2 caled m / z = 744.36 amu; found [M + H]⁺ = 745.39, [M + Na]⁺ = 777.32

253

Example 96

<img file="MX368258B_D0150.tif" />

Chemical Formula: C3₆H₅₅N₅OR₆S

Exact Mass: 685.39

Molecular Weight: 685.92 (96) (S, E) -4 - ((S) -2 - ((S) -3- (4- (2-aminoethoxy) phenyl) -3-methyl-2 (methylamino) butanamide) -N, 3,3-trimethylbutanamido) -N (benzylsulfonyl) -2,5-dimethylhex-2-enamide

The title compound was prepared as follows: Et3N (4 eq) was added to a cold solution (0^or C) under stirring of MsCl (3.7 eq) in CH2CI2. After 2 min, a solution of Boc-protected Example 94 in CH was added₂C1₂. The pale yellow mixture was stirred cold for 5 min and then at room temperature for 72 h. The resulting mixture was diluted with EtOAc and washed successively with 1M citric acid (Ix), NaHCO₃ 1M (Ix) and brine (lx). The organic compounds were dried over MgSO₄, filtered and concentrated in vacuo to provide the mesylated alcohol (HPLC / MS - [M + Na] ⁺ = 887.42) which was used in the next stage without further purification.

The mesylate was dissolved in DMF and treated with NaN₃ (7 eq). The resulting suspension was stirred at room temperature for

254 h and then at 60 ⁰ C for 5 h. The reaction mixture was diluted with H₂0, acidified with 1M HC1 and extracted with CH₂C1₂ (4x). The combined organic extracts were dried over MgSO₄, filtered and concentrated in vacuo to provide the azido product (HPLC / MS - [M + Na] ⁺ = 834.44) which was used in the next stage without further purification.

The azide was dissolved in THF / H₂Or (10: 1) and treated with tributylphosphine (3.5 eq). The mixture was stirred at room temperature for 21 h and then concentrated in vacuo. The resulting residue was dissolved in EtOAc and washed successively with 1M HC1 (3x), NaHCO₃ 1M (3x), H₂O (2x) and brine (2x). The organic compounds were dried over MgSO₄, filtered, concentrated in vacuo and purified by silica gel column chromatography (eluting with MeOH / CH mixtures₂C1₂) to give the primary amine as a white solid (HPLC / MS - [Μ + H] ⁺ = 786.45).

The amine dissolved in CH₂C1₂ and treated with TFA. After stirring for 1 h, the reaction mixture was concentrated in vacuo. The off-white solid residue was dissolved in the minimum amount of MeOH, cooled to 0^or C and treated with ether to precipitate the desired diamine product as an off-white solid with a yield of 6% over

255 Four synthetic steps

NMR (400 MHz, Methanol-dJ δ 7.50 (d, J = 8.6 Hz, 2H), 7.37 (s, 5H), 7.09 (d, J = 8.6 Hz, 2H), 6.41 (d, J = 9.4 Hz, 1H ), 5.02 (t, J = 10.0 Hz, 1H), 4.91 (s, 1H), 4.70 (s, 2H), 4.27 (t, J = 5.0 Hz, 2H), 3.40 (t, J = 5.0 Hz, 2H ), 3.37 (s, 1H), 3.12 (s, 3H), 2.47 (s, 3H), 2.06 - 1.95 (m, 1H), 1.94 (d, J = 1.4 Hz, 3H), 1.45 (s, 3H) , 1.37 (s, 3H), 1.08 (s, 9H), 0.89 (dd, J-9.7, 6.6 Hz, 6H).

C₃8H₅₅N₅OR₆S caled m / z = 685.39 amu; found [M + H]⁺ = 686.32, [M + Na]⁺ = 708.27, [(M + 2H) / 2]²⁺ = 343.77

Example 97

OR

<img file="MX368258B_D0151.tif" />

Chemical F69fli) la: C₃5H4₈F3N₅OR₆S

Exact Mass: 723.33 (S, E) -2,5-MbleoeltaíWei ^ t :( 723 (85, 2,2-trifluoroacetamido) phenylsulfonyl) -4 - ((S) -N, 3,3-trimethyl-2 ((S) -3-methyl-2- (methylamino) -3-phenylbutanamido) butanamido) hex-2-enamide

The title compound was prepared according to Example 3 and the

2,2,2-Trifluoro-N- (2-sulfamoylphenyl) acetamide according to General Procedures 2, and 7.

^XH NMR (400 MHz, Methanol-d₄) δ 8.27 (d, J = 8.4 Hz, 1H), 8.05

256 (d, J = 7.8 Hz, 1H), 7.67 (t, J = 7.9 Hz, 1H), 7.54 (d, J =

8.1 Hz, 2H), 7.48 (t, J = 7.7 Hz, 2H), 7.40 (dt, J = 13.3,

7.4 Hz, 2H), 6.57 (d, J = 9.2 Hz, 1H), 4.92 (s, 2H), 4.34 (s,

1H), 3.17 (s, 3H), 2.50 (s, 3H), 2.06 (m, 1H), 1.87 (d, J =

1.3 Hz, 3H), 1.45 (s, 3H), 1.33 (s, 3H), 1.07 (s, 9H), 0.91 (dd, J = 6.6, 3.5 Hz, 6H).

¹⁹F NMR (377 MHz, Methanol-d₄) δ -76.96, -77.73.

CssH ^ FaNsOgS caled m / z = 723.33 amu; found [M + H]⁺ = 723.34, [M + Na]⁺ = 746.23

Example 98

<img file="MX368258B_D0152.tif" />

Chemical Formula: O33N49N5O5S

Exact Mass: 627.35

Molecular weight: . 627.84 (98) (S, E) -N- (2-aminophenylsulfonyl) -2,5-dimethyl-4 - ((S) -N, 3,3-trimethyl-2 - ((S) -3-methyl-2- (methylamino) -3phenylbutanamide) butanamide) hex-2-enamide

The title compound was prepared according to Example 3 and 2,2,2-trifluoro-N- (2-sulfamoiiphenyl) acetamide according to General Procedures 2, 3 and 7.

^TH NMR (400 MHz, Methanol-d₄) δ 7.75 (dd, J = 8.2, 1.5 Hz, 1H),

257

7.55 (d, J = 7.8 Hz, 2H), 7.48 (t, J = 7.7 Hz, 2H), 7.38 (t, J = 7.4 Hz, 1H), 7.33 - 7.27 (m, 1H), 6.81 (d, J = 8.2 Hz, 1H), 6.69 (t, J = 7.5 Hz, 1H), 6.49 (dd, J = 9.1, 1.5 Hz,

1H), 4.97 (t, J = 10.1 Hz, 1H), 4.92 (s, 1H), 4.35 (s, 1H),

3.17 (s, 3H), 2.51 (s, 3H), 2.07 (m, 1H), 1.88 (d, J = 1.4

Hz, 3H), 1.46 (s, 3H), 1.36 (s, 3H), 1.06 (s, 9H), 0.92 (t, J = 6.8 Hz, 6H).

CssH ^ gNsOsS caled m / z = 627.35 amu; found [M + H]⁺ = 628.36, [M + Na]⁺ = 650.37, [(M + 2H) / 2]²⁺ = 314.76

Example 99

Chemical formula:

Exact Mass:

Molecular weight:

C39H52N4O5S

688.37

688.92

<img file="MX368258B_D0153.tif" />

(99) (S, E) -N- (biphenyl-4-ylsulfonyl) -2,5-dimethyl-4 - ((S) -N, 3,3 trimethyl-2 - ((S) -3-methyl- 2- (methylamino) -3 phenylbutanamide) butanamide) hex-2-enamide

The title compound is prepared using Example 56 protected with Boc with phenylboronic acid according to General Procedures 8 and 7.

^XH NMR (400 MHz, Methanol-d₄) δ 8.12 (d, J = 8.3 Hz, 2H), 7.83

258

<td>(d, J =</td><td> 8.4</td><td>Hz, 2H</td><td> ), 7.71</td><td>(d, J = 7.7</td><td>Hz, 2H), 7.52</td><td>(dd,</td><td>J =</td>

<td> 11.6, 7.</td><td>6 Hz</td><td>, 4H),</td><td>7.45 (t</td><td colspan="2">, J = 7.3 Hz, 3H), 7.36 (t</td><td>, J =</td><td> = 7.2</td>

<td>Hz, 1H),</td><td colspan="2">6.52 (d,</td><td>J = 9.4</td><td>Hz, 1H), 4.</td><td>96 (t, J = 9.5</td><td>Hz,</td><td>1 HOUR) ,</td>

<td>4.92 (s,</td><td>1 HOUR)</td><td> , 4.33</td><td>(s, 1H)</td><td>, 3.18 (s,</td><td>3H), 2.50 (s,</td><td>3H),</td><td> 2.14</td>

<td> - 2.03</td><td>(m,</td><td>1H), 1</td><td>.88 (s,</td><td>3H), 1.45</td><td>(s, 3H), 1.35</td><td>(s,</td><td>3H),</td>

<td>1.07 (s,</td><td>9H)</td><td> , 0.92</td><td>(t, J =</td><td>6.9 Hz, 6H)</td><td></td><td></td><td></td>

C39H52N4O5S caled m / z = 688.37 amu; found [M + H]⁺ = 689.10, [M + Na]⁺ = 711.32

Example 100

<img file="MX368258B_D0154.tif" />

Chemical Formula: C39H53N5O5S

<img file="MX368258B_D0155.tif" />

NH₂

Exact Mass: 703.38

Molecular Weight: 703.93 (100) (S, E) -N- (4'-aminobiphenyl-4-ylsulfonyl) -2,5-dimethyl-4 - (($) -

N, 3,3-trimethyl-2 - ((S) -3-methy1-2- (methylamino) -3phenylbutanamido) butanamide) hex-2-enamide

The title compound was prepared from Example 68 protected with Boc with 4- (tertbutoxycarbonylamino) phenylboronic acid according to General Procedures 8 and 7.

^XH NMR (400 MHz, Methanol-d₄) δ 8.05 (d, J = 8.6 Hz, 2H), 7.75

259 (d, J = 8.6 Hz, 2H), 7.59 - 7.51 (m, 4H), 7.45 (t, J = 7.7 Hz, 2H), 7.36 (t, J = 7.3 Hz, 1H), 6.91 (d, J = 8.3 Hz, 2H),

6.50 (d, J = 9.1 Hz, 1H), 4.98 - 4.92 (m, 1H), 4.91 (s, 1H),

4.34 (s, 1H), 3.18 (s, 3H), 2.50 (s, 3H), 2.13 - 2.03 (m,

1H), 1.88 (d, J = 1.4 Hz, 3H), 1.45 (s, 3H), 1.35 (s, 3H),

1.06 (s, 9H), 0.92 (t, J = 6.2 Hz, 6H).

C39H53N5O5S caled m / z = 703.38 amu; found [M + H]⁺ = 704.26, [M + Na]⁺ = 726.41, [(M + 2H) / 2]²⁺ = 352.77

Example 101 (101) (S, E) -N- (4-fluorobenzyl sulfonyl) -2,5-dimethyl-4 - ((S) -N, 3,3 trimethyl-2 - ((S) -3-methyl- 2- (methylamino) -3 phenylbutanamide) butanamide) hex-2-enamide

The title compound was prepared according to Example 3 and 4-fluorobenzyl sulfonamide according to General Procedures 2 and 7.

^ΤΗ NMR (400 MHz, Methanol-d4) δ 7.60 - 7.52 (m, 2H), 7.48 (t,

J = 7.7 Hz, 2H), 7.44 - 7.34 (m, 3H), 7.18 - 7.05 (m, 2H),

6.41 (dd, J = 9.5, 1.7 Hz, 1H), 5.06 (t, J = 10.0 Hz, 1H),

4.94 (s, 1H), 4.74 (s, 2H), 4.35 (s,

1H), 3.13 (s, 3H), 2.51

260 (s, 3H), 2.07 - 1.97 (m, 1H), (s, 3H), 1.39 (s, 3H), 1.09

6H).

1.95 (d, J = 1.4 Hz, 3H), 1.48 (s, 9H), 0.90 (t, J = 6.3 Hz,

C34H49FN4O5S caled m / z = 644.34 found [M + H]⁺ = 645.32

Example 102

<img file="MX368258B_D0156.tif" />

(102) (S, E) -2,5-dimethyl-N- (3- (trifluoromethyl) benzyl sulfonyl) -4 ((S) -N, 3,3-trimethyl-2 - ((S) -3-methyl -2- (methylamino) -3phenylbutanamido) butanamide) hex-2-enamide

The title compound was prepared according to Example 3 and the

3-trifluorobenzyl sulfonamide according to the Procedures

<td rowspan="2">General 2^XH NMR (400</td><td colspan="9">and 7.</td>

<td colspan="3">MHz, Methanol-d4</td><td colspan="2">) δ 7.74 - 7</td><td>. 64 (m,</td><td>3H), 7</td><td> . 61</td><td>(d,</td>

<td>J = 7.7 Hz,</td><td>1 HOUR) ,</td><td> 7.60</td><td> - 7.</td><td>54 (m</td><td>, 2H),</td><td>7.48 (t</td><td>, J =</td><td> 7.7</td><td>Hz,</td>

<td>2H), 7.38 (</td><td>: t, j =</td><td> ; 7.3</td><td>Hz,</td><td>1 HOUR) ,</td><td> 6.42 (</td><td>dd, J =</td><td> 9.4,</td><td> 1.7</td><td>Hz,</td>

<td>1H), 5.06 (</td><td>t, j -</td><td> 10.0</td><td>Hz,</td><td>1 HOUR) ,</td><td> 4.93 (</td><td>s, 1H),</td><td> 4.36 (</td><td>s,</td><td>1 HOUR) ,</td>

<td>3.13 (s, 3H</td><td> ), 2.51</td><td>(s,</td><td>3H),</td><td> 2.07</td><td> - 1.97</td><td>(m, 1H),</td><td> , 1.95</td><td>(d,</td><td>J =</td>

<td>1.4 Hz, 3H)</td><td> , 1.48</td><td>(s,</td><td>3H),</td><td> 1.39</td><td>(s, 3H</td><td> ), 1.08</td><td>(s, 9H</td><td> )</td><td> 0.89</td>

(d, J = 6.5 Hz, 6H).

C35H4 9 F3N4O5S caled m / z = 694.34 found [M + H]⁺ = 695.38

261

Example 103

<img file="MX368258B_D0157.tif" />

(103) (S, E) -2,5-dimethyl-N- (3- (trifluoromethoxy) benzylsulfonyl) -4 ((S) -N, 3,3-trimethyl-2 - ((S) -3-methyl -2- (methylamino) -3phenylbutanamido) butanamide) hex-2-enamide

The title compound was prepared according to Example 3 and 3-trifluoromethoxybenzyl sulfonamide according to General Procedures 2 and 7.

NMR (400 MHz, Methanol-d4) δ 7.56 (d, J = 7.8 Hz, 2H), 7.48

<td>(t,</td><td colspan="2">J = 7.9 Hz, 3H),</td><td>7.43 - 7.36 (m,</td><td>2H), 7.32 (d,</td><td>J =</td><td> 9.3</td>

<td>Hz,</td><td>2H), 6.43 (dd,</td><td>J</td><td>= 9.4, 1.7 Hz,</td><td>1H), 5.06 (t,</td><td></td><td> 10.0</td>

<td>Hz,</td><td>1H), 4.93 (s,</td><td>1 HOUR)</td><td>, 4.82 (s, 2H),</td><td>4.35 (s, 1 H),</td><td> 3.13</td><td>(s,</td>

<td>3H),</td><td>2.51 (s, 3H),</td><td> 2.</td><td>07 - 1.97 (m, 1H</td><td>), 1.95 (d, J =</td><td> = 1.4</td><td>Hz,</td>

<td>3H),</td><td>1.48 (s, 3H),</td><td> 1.</td><td>39 (s, 3H), 1.08</td><td>(S, 9H), 0.90</td><td>(dd,</td><td>J =</td>

<td> 6.6,</td><td>4.3 Hz, 6H).</td><td></td><td></td><td></td><td></td><td></td>

C35H49F3N4O6S caled m / z = 710.33 found [M + H]⁺ = 711.38

262

Example 104

<img file="MX368258B_D0158.tif" />

NH

N

OR

<img file="MX368258B_D0159.tif" />

Cl

Cl (104) (S, E) -N- (3,4-dichlorobenzyl sulfonyl) -2,5-dimethyl-4 - ((S) -N, 3,3 trimethyl-2 - ((S) -3-methyl -2- (methylamino) -3 phenylbutanamido) butanamide) hex-2-enamide

The title compound was prepared according to Example 3 and 3,4-dichlorobenzyl sulfonamide according to General Procedures 2, and 7.

^XH NMR (400 MHz, Methanol-d4) δ 7.56 (td, J = 5.2, 4.5, 1.9 Hz, 4H), 7.48 (t, J = 7.7 Hz, 2H), 7.38 (t, J = 7.3 Hz, 1H) , 7.33 (dd, J = 8.4, 2.1 Hz, 1H), 6.41 (dd, J = 9.5, 1.8 Hz,

1H), 5.06 (t, J = 10.0 Hz, 1H), 4.93 (s, 1H), 4.77 (s, 2H),

4.36 (s, 1H), 3.14 (s, 3H), 2.52 (s, 3H), 2.07 - 1.97 (m,

1H), 1.95 (d, J = 1.4 Hz, 3H), 1.49 (s, 3H), 1.39 (s, 3H),

1.08 (s, 9H), 0.90 (dd, J = 6.6, 4.9 Hz, 6H).

C34H48CI2N4O5S caled m / z = 694.27 found [M + H]⁺ = 695.32

263

Example 105

<img file="MX368258B_D0160.tif" />

(105) (S, E) -N- (2-cyanobenzyl sulfonyl) -2,5-dimethyl-4 - ((S) -N, 3,3-trimethyl-2 - ((S) -3-methyl-2- ( methylamino) -3phenylbutanamide) butanamide) hex-2-enamide

The title compound was prepared according to Example 3 and the

2-cyanobenzyl sulfonamide according to the Procedures

General 2, and 7.

<td>^XH NMR</td><td colspan="2">(400 MHz, Methanol-d₄)</td><td>δ 7.81</td><td>(dd.</td><td>, J = 7.</td><td> 7,</td><td> 1.3</td><td>Hz,</td><td>1 HOUR)</td>

<td> 7.72 (</td><td>td, J =</td><td>= 7.7, 1.3 Hz,</td><td>1 HOUR) ,</td><td> 7.66</td><td>(d, J</td><td></td><td> 7.7</td><td>Hz,</td><td>1 HOUR)</td>

<td> 7.62 -</td><td> 7.59 (</td><td>m, 1H), 7.58</td><td> - 7.53</td><td>(m,</td><td>2H), 7.</td><td> 48</td><td>(t,</td><td>J =</td><td> 7 .</td>

<td>Hz, 2H</td><td> ), 7.38</td><td>(t, J = 7.3 H</td><td>z, 1H),</td><td colspan="2">6.50 (d, J</td><td> =</td><td> 9.4</td><td>Hz,</td><td>1 HOUR)</td>

<td> 5.08 (</td><td>dd, J =</td><td>10.6, 9.3 Hz,</td><td>1H), 4</td><td> . 99</td><td>(s, 2H),</td><td> , 4</td><td> . 95</td><td>(s,</td><td>1 HOUR)</td>

<td> 4.36 (</td><td>s, 1H),</td><td>3.16 (s, 3H)</td><td> , 2.52</td><td>(s,</td><td>3H), 2</td><td> .09</td><td> -</td><td> 1.99</td><td>(m</td>

<td>1H), 1</td><td>.98 (d,</td><td>J = 1.4 Hz,</td><td>3H), 1.</td><td> 49</td><td>(s, 3H),</td><td> 1,</td><td> .39</td><td>(s,</td><td>3H)</td>

<td> 1.10 (</td><td>s, 9H),</td><td>0.94 (d, J =</td><td>6.6 Hz,</td><td>3H)</td><td> , 0.91</td><td>(d,</td><td>J =</td><td> = 6.6</td><td>Hz</td>

3H).

C₃₅H4₉N₅OR₅S caled m / z = 651.35 found [M + H]⁺ = 652.38

264

Example 106

<img file="MX368258B_D0161.tif" />

<img file="MX368258B_D0162.tif" />

(106) (S, E) -N- (3-Chlorobenzyl sulfonyl) -2,5-dimethyl-4 - ((S) -N, 3,3-trimethyl-2 - ((S) -3-methyl-2- ( methylamino) -3phenylbutanamide) butanamide) hex-2-enamide

The title compound was prepared according to Example 3 and 3-chlorobenzyl sulfonamide according to General Procedures 2 and 7.

And NMR (400 MHz, Methanol-d4) δ 7.58 - 7.53 (m, 2H), 7.48 (t,

J = 7.6 Hz, 2H), 7.43 - 7.34 (m, 4H), 7.32 (d, J = Ί.5 Hz,

<td>1 HOUR) ,</td><td>6.42 (d,</td><td>J =</td><td>9.5 Hz, 1H),</td><td>5.06 (t, J =</td><td> 10.0</td><td>HZ,</td><td>1 HOUR) ,</td>

<td> 4.94</td><td>(s, 1H),</td><td> 4.74</td><td>(s, 2H), 4.33</td><td>(s, 1H), 3.13</td><td>(s,</td><td>3H),</td><td> 2.50</td>

<td>(s,</td><td>3H), 2.07</td><td> - 1.</td><td>97 (m, 1 H), 1.</td><td>95 (d, J = 1.4</td><td>Hz,</td><td>3H),</td><td> 1.48</td>

<td>(s,</td><td>3H), 1.39</td><td>(s,</td><td>3H), 1.08 (s,</td><td>9H), 0.90 (t,</td><td>J </td><td> = 7.2</td><td>Hz,</td>

6H).

C34H49CIN4O5S caled m / z = 660.31 found [M + H]⁺ = 661.32

265

<img file="MX368258B_D0163.tif" />

Example 107

<img file="MX368258B_D0164.tif" />

c \ N * H

NH₂ (107) (S, E) -N- (4-amino-2-ethylphenylsulfonyl) -2,5-dimethyl-4 - ((S) -

N, 3,3-trimethyl-2 - ((S) -3-methyl-2- (methylamino) -3phenylbutanamido) butanamido) hex-2-enamide

The title compound was prepared according to Example 3 and 2-ethylbenzyl sulfonamide according to General Procedures 2 and 7.

^XH NMR (400 MHz, Methanol-d₄) δ 7.79 (d, J = 8.7 Hz, 1H), 7.55

<td>(d,</td><td>J = 7.9 Hz, 2H), 7.</td><td>48 (t,</td><td>J =</td><td> 7.6</td><td>Hz,</td><td>2H), 7.37</td><td>(t,</td><td>J =</td>

<td> 7.4</td><td>Hz, 1H), 6.57 (d, J</td><td> = 2.3</td><td>Hz,</td><td>1 HOUR) ,</td><td> 6.54</td><td>(dd, J =</td><td> 8.8,</td><td> 2.4</td>

<td>Hz,</td><td>1H), 6.46 (d, J = 9.</td><td>.4 Hz,</td><td>1 HOUR) ,</td><td> 5.01</td><td>(t,</td><td>J = 10.0</td><td>Hz,</td><td>1 HOUR) ,</td>

<td> 4.92</td><td>(s, 1H), 4.34 (s,</td><td>1 HOUR) ,</td><td> 3.16</td><td>(s,</td><td>3H),</td><td> 2.99 -</td><td> 2.90</td><td>(m,</td>

<td>2H),</td><td>2.50 (s,</td><td>3H),</td><td> 2.11 -</td><td> 2.00</td><td>(m, 1H),</td><td> 1.87</td><td>(d, J = 1.4 Hz,</td>

<td>3H),</td><td>1.47- (s,</td><td>3H),</td><td> 1.38</td><td>(s, 3H</td><td> ), 1.22</td><td>(t, J</td><td>= 7.5 Hz, 3H),</td>

<td> 1.06</td><td>(s, 9H),</td><td> 0.91</td><td>(dd, J</td><td> = 6.6</td><td>Hz, 6H).</td><td></td><td></td>

C35H53N5O5S caled m / z = 655.38 found [M + H]⁺ = 656.4

266

Example 108

<img file="MX368258B_D0165.tif" />

(108) (S, E) -N- (4-amino-3- (trifluoromethoxy) phenylsulfonyl) -2,5-dimethyl-4 - ((S) -N, 3,3-trimethyl-2 - ((S) - 3-methyl-2- (methylamino) 3-phenylbutanamido) butanamido) hex-2-enamide

The title compound was prepared according to Example 3 and the

2,2,2-trifluoro-N- (4-sulfamoyl-2- (trifluoromethoxy) phenyl) acetamide according to

General Procedures 2, 3 and 7.

<td>^XH NMR (400</td><td>MHz, Methanol-d₄;</td><td>l δ</td><td> 7.81 -</td><td>7.75 (m, 1 H), 7.</td><td> 71</td><td>(dd,</td>

<td>J = 8.7, 2.</td><td>1 Hz, 1H), 7.55</td><td>(d,</td><td>J = 1.</td><td>9 Hz, 2H), 7.47</td><td>(t,</td><td>J =</td>

<td>7.6 Hz, 2H)</td><td>, 7.37 (t, J =</td><td> 7.1</td><td>Hz, ih;</td><td>I, 6.89 (d, J =</td><td> 8.7</td><td>Hz,</td>

<td>1H), 6.51 -</td><td>6.42 (m, 1 H), 4</td><td> . 98</td><td>(t, J =</td><td>= 10.0 Hz, 1H), 4</td><td> . 92</td><td>(t,</td>

<td>J = 4.1 Hz,</td><td>. 1H), 4.37 (s,</td><td>1 HOUR)</td><td> , 3.16</td><td>(s, 3H), 2.51 (</td><td>s,</td><td>3H),</td>

<td> 2.12 - 2.01</td><td>(m, 1H), 1.88 1</td><td>(d,</td><td>J = 1.4</td><td>Hz, 3H), 1.47 (</td><td>S,</td><td>3H),</td>

<td>1.37 (s, 3H</td><td>), 1.07 (s, 9H),</td><td>0. S</td><td>) 2 (dd,</td><td>J = 6.6 Hz, 6H).</td><td></td><td></td>

<td>C34H48F3N5O6S</td><td>caled m / z = 711.</td><td> , 33</td><td colspan="2">found [M + H]⁺ = 712</td><td> . 4</td><td></td>

267

Example 109

<img file="MX368258B_D0166.tif" />

(109) (S, E) -N- (4-amino-2,3-dimethylphenylsulfonyl) -2,5-dimethyl-4 ((S) -N, 3,3-trimethyl-2 - ((S) - 3-methyl-2- (methylamino) -3phenylbutanamido) butanamido) hex-2-enamide

The title compound was prepared according to Example 3 and the

2,2,2-Trifluoro-N- (4-sulfamoyl-2,3-dimethylphenyl) acetamide

<td>agreement</td><td>to the Procedures</td><td colspan="2">General 2,</td><td>3 and 7.</td><td></td><td></td>

<td>^XH NMR</td><td>(400 MHz, Methanol-d₄)</td><td>δ</td><td>7.75 (d, J</td><td> = 8.8</td><td>Hz, 1H),</td><td> 7.55</td>

<td>(d, J =</td><td>7.9 Hz, 2H), 7.47 (</td><td>t</td><td>J = 7.7 Hz</td><td>, 2H),</td><td>7.37 (t,</td><td>J =</td>

<td>6.9 Hz,</td><td>1H), 6.63 (d, J = 8</td><td> . 8</td><td>Hz, 1H), 6</td><td>.46 (d</td><td>, J = 9.7</td><td>Hz,</td>

<td>1H), 5.</td><td>00 (t, J = 10.0 Hz,</td><td>1 HOUR)</td><td>P 4.93 (s,</td><td>1 HOUR) ,</td><td>4.32 (s,</td><td>1 HOUR) ,</td>

<td>3.17 (s</td><td>, 3H), 2.54 (s, 3H),</td><td> 2.</td><td>49 (s, 3H),</td><td> 2.09</td><td>(s, 3H),</td><td> 2.08</td>

<td> - 2.02</td><td>(m, 1H), 1.87 (d, J -</td><td> = 1</td><td>.4 Hz, 3H),</td><td> 1.47</td><td>(s, 3H),</td><td> 1.37</td>

<td>(s, 3H)</td><td>, 1.07 (s, 9H), 0.92</td><td>(from</td><td>ί, J = 6.8,</td><td>6.5 Hz</td><td>, 6H).</td><td></td>

C35H53N5O5S caled m / z = 655.38 found [M + H]⁺ = 656.4

268

Example 110

<img file="MX368258B_D0167.tif" />

(110) (S, E) -N- (4-amino-5,6,7,8-tetrahydronaphthalen-1-ylsulfonyl) 2,5-dimethyl-4 - ((S) -N, 3,3-trimethyl -2 - ((S) -3-methyl-2 (methylamino) -3-phenylbutanamido) butanamido) hex-2-enamide

The title compound was prepared according to Example 3 and the

2,2,2-Trifluoro-N- (4-sulfamoyl-5,6,7,8-tetrahydronaphthalen-l-yl) acetamide according to General Procedures 2, 3 and

7.

* H NMR (400 MHz, Methanol-d₄) δ 7.74 (d, J = 8.7 Hz, 1H), 7.55

<td>(d,</td><td>J = 7.9 Hz, 2H),</td><td> 7.48</td><td>(t, J = 7.6 Hz, 2H),</td><td>7.38 (t,</td><td>J =</td>

<td> 7.2</td><td>Hz, 1H), 6.60 (d,</td><td>J =</td><td>8.7 Hz, 1H), 6.46 (d</td><td>, J = 9.2</td><td>Hz,</td>

<td>1 HOUR) ,</td><td>5.00 (t, J = 10.</td><td>0 Hz,</td><td>1H), 4.95-4.91 (m,</td><td>1H), 4.36</td><td>(s,</td>

<td>1 HOUR) ,</td><td>3.17 (s, 3H), 3</td><td> .10 -</td><td>- 3.05 (m, 2H), 2.51</td><td>(s, 3H),</td><td> 2.46</td>

<td>(t,</td><td>J = 6.5 Hz, 2H),</td><td> 2.10</td><td>- 2.02 (m, 1H), 1.88</td><td>(s, 3H),</td><td> 1.87</td>

<td> - 1.</td><td>75 (m, 4H), 1.47</td><td>(s,</td><td>3H), 1.38 (s, 3H),</td><td>1.07 (s,</td><td>9H),</td>

<td> 0.92</td><td>(dd, J = 7.1 Hz,</td><td>6H).</td><td></td><td></td><td></td>

<td colspan="2">C37H55N5O5S caled m / z =</td><td> 681.</td><td>39 found [M + H]⁺ =</td><td> 682.4</td><td></td>

269

Example 111

N

<img file="MX368258B_D0168.tif" />

NH₂ (111) (S, E) -N- (4-amino-3-methylphenylsulfonyl) -2,5-dimethyl-4 - ((S) N, 3,3-trimethyl-2 - ((S) -3- methyl-2- (methylamino) -3phenylbutanamido) butanamido) hex-2-enamide

The title compound was prepared according to Example 3 and the

2,2,2-Trifluoro-N- (2-methyl-4-sulfamoylphenyl) acetamide according to General Procedures 2, 3 and 7.

^XH NMR (400 MHz, Methanol-d₄) δ 7.64 (s, 1H), 7.61 (dd, J =

8.5, 2.3 Hz, 1H), 7.57 - 7.51 (m, 2H), 7.48 (t, J = 7.7 Hz, 2H), 7.41 - 7.35 (m, 1H), 6.71 (d, J = 8.5 Hz, 1H), 6.43 (dd,

J = 9.3, 1.6 Hz, 1H), 4.96 (t, J = 10.0 Hz, 1H), 4.92 (s,

1H), 4.35 (s, 1H), 3.16 (s, 3H), 2.51 (s, 3H), 2.17 (s, 3H),

2.10 - 2.01 (m, 1H), 1.87 (d, J = 1.4 Hz, 3H), 1.46 (s, 3H),

1.36 (s, 3H), 1.07 (s, 9H), 0.91 (dd, J = 6.3 Hz, 6H).

C₃₄H₅iN₅O5S caled m / z = 641.36 found [M + H]⁺ = 642.4

270

Example 112

<img file="MX368258B_D0169.tif" />

<img file="MX368258B_D0170.tif" />

F nh₂ (112) (S, E) -N- (4-amino-3-fluorophenylsulfonyl) -2,5-dimethyl-4 - ((S) N, 3,3-trimethyl-2 - ((S) -3- methyl-2- (methylamino) -3phenylbutanamido) butanamido) hex-2-enamide

The title compound was prepared according to Example 3 and the

2,2,2-Trifluoro-N- (2-fluoro-4-sulfamoylphenyl) acetamide according to General Procedures 2, 3 and 7.

^XH NMR (400 MHz, Methanol-d₄) δ 7.62 - 7.55 (m, 3H), 7.54 (s, 1H), 7.48 (t, J = 7.7 Hz, 2H), 7.37 (t, J - 7.3 Hz, 1H), 6.85 (t, J = 8.6 Hz , 1H), 6.45 (d, J = 9.3 Hz, 1H), 4.98 (t, J = 9.9 Hz, 1H), 4.92 (s, 1H), 4.34 (s, 1H), 3.16 (s, 3H), 2.50 (s, 3H), 2.12 - 2.00 (m, 1H), 1.88 (d, J = 1.4 Hz, 3H), 1.46 (s, 3H), 1.37 (s, 3H), 1.07 (s, 9H), 0.91 ( dd, J = 6.8 Hz, 6H).

C₃3H₄₈FN₅OR₅S caled m / z = 645.34 found [M + H]⁺ = 646.4

271

Example 113

<img file="MX368258B_D0171.tif" />

(113) (S, E) -N- (4-amino-3-ethylphenylsulfonyl) -2,5-dimethyl-4- ((S) -

N, 3,3-trimethyl-2 - ((S) -3-methyl-2- (methylamino) -3phenylbutanamido) butanamido) hex-2-enamide

The title compound was prepared according to Example 3 and the

2,2,2-trifluoro-N- (2-ethyl-4-sulfamoylphenyl) acetamide according to Generic Procedures 2, 3 and 7.

NMR (400 MHz, Methanol-d₄) δ 7.66 (d, J = 2.3 Hz, 1H), 7.61 (dd, J = 8.6, 2.3 Hz, 1H), 7.55 (d, J = 7.6 Hz, 2H), 7.48 (t, J = 7.7 Hz, 2H ), 7.37 (t, J = 7.3 Hz, 1H), 6.71 (d, J = 8.5

<td>Hz,</td><td>1 HOUR)</td><td> , 6.</td><td>43 (dd</td><td>, J =</td><td> 9.3, 1.7</td><td>Hz, 1H), 4.96 (</td><td>t, J = 9.9</td><td>Hz,</td>

<td>1 HOUR) ,</td><td> 4 .</td><td> 92 (</td><td>s, 1H)</td><td colspan="2">, 4.35 (s, 1H)</td><td>, 3.16 (s, 3H),</td><td>2.54 (dd,</td><td>J =</td>

<td> 7.4,</td><td> 2.</td><td>2 Hz</td><td>, 2H),</td><td> 2.51</td><td>(s, 3H),</td><td>2.12 - 1.99 (m,</td><td>1H), 1.87</td><td>(d,</td>

<td>J =</td><td> 1.4</td><td>Hz,</td><td>3H),</td><td> 1.46</td><td>(s, 3H),</td><td>1.36 (s, 3H), 1.</td><td>27 (t, J =</td><td> 7.5</td>

<td>Hz,</td><td>3H)</td><td> , 1.</td><td>07 (s,</td><td>9H),</td><td>0.91 (dd</td><td>, 1 = 6.4 Hz, 6H</td><td></td><td></td>

C35H53N5O5S caled m / z = 655.38 found [M + H]⁺ = 656.5

272

Example 114

<img file="MX368258B_D0172.tif" />

(114) (S, E) -N- (4-amino-3- (trifluoromethyl) phenylsulfonyl) -2.5 dimethyl-4 - ((S) -N, 3,3-trimethyl-2 - ((S) -3-methyl-2- (methylamino)

3-phenylbutanamide) butanamide) hex-2-enamide

The title compound was prepared according to Example 3 and the

2,2,2-Trifluoro-N- (2-trifluoromethyl-4- sulfamoylphenyl) acetamide according to the Procedures

General 2, 3 and 7.

^ΤΗ NMR (400 MHz, Methanol-d₄) δ

8.04 (s, 1H)

7.87 (d

8.8

Hz, 1H), 7.55 (d,

7.6 Hz

2H),

7.48 (t,

7.3 Hz, 2H)

7.36 (dd

14.5

7.4

Hz, 1H),

6.89 (d

8.9 Hz, 1H)

6.47 (d, J = 9.3 Hz

1 HOUR)

4.99 (t

J = 10.2

Hz, 1H), 4.92 (s

1 HOUR)

1 HOUR)

3.16

3H),

2.50 (s

3H), 2.11 - 2.00

1H), 1.88

3H), 1.47 (s, 3H), 1.37

3H), 1.07 (s

9H),

0.91 (dd

7.0 Hz, 6H).

C₃₄H₄₈F₃N₅O5S caled m / z = 695.33 found [M + H]

696.4

273

Example 115

<img file="MX368258B_D0173.tif" />

(115) (S) -1-Isopropyl-N - ((S) -1- (((S, E) -6- (3mercaptopropylsulfonamido) -2,5-dimethyl-6-oxohex-4-en-3il) (methyl) amino) -3,3-dimethyl-l-oxobutan-2-yl) piperidine-2-carboxamide

To a solution of (S, E) -ethyl 4 - ((S) -2- (tertbutoxycarbonylamino) -N, 3,3-trimethylbutanamide) -2,5-dimethylhex-2-methyl enoate (0.373g, 0.905mmol) in CH2CI2 (5 ml) trifluoroacetic acid (2 ml) was added. The reaction was monitored by HPLC and after complete conversion of the starting material was concentrated under reduced pressure. The N-isopropyl-pipecolic acid (0.200g, 1.3 equiv) was dissolved in CH₂C1₂ (5 mi) and stirred at 0^or C, to which HBTU (0.450 g, 1.3 equiv) and N, N-diisopropylethylamine (0.400uL, 2.5 equiv) was added. After 10 minutes, the above unprotected dipeptide was added as a solution in CH₂C1₂ (~ 1 mi). The reaction was monitored by HPLC until complete consumption of the dipeptide at which time the entire reaction was concentrated under reduced pressure. The crude reaction mixture dissolved.

274 in CH2CI2 and purified by silica gel chromatography (1-20% MeOH (5% NH₄OH) in CH₂C1₂) .

The resulting ester is saponified with LiOH in 1,4-dioxane. The resulting carboxylic acid (0.128 g, 0.29 mmol) was dissolved in CH₂C1₂ (5 ml) and to the stirred solution was added dicyclohexylcarbodiimide (0.084 g, 1.4 equiv), N, N-dimethylaminopyridine (0.05 g, 1.4 equiv) and 3 (tritylthio) propan-l-sulfonamide (0.174 g, 1.5 equiv). The resulting mixture was stirred overnight and its reaction progress was monitored by HPLC-MS. When the reaction was complete, the mixture was concentrated under reduced pressure and the residue was purified by silica gel chromatography (5-30% MeOH in CH₂C1₂) to give the S-trityl derivative of the original compound as a colorless oil (0.056 g).

^TH NMR (400 MHz, Methanol-d₄) δ 7.44 - 7.35 (m, 6H), 7.36 -

<td> 7.15</td><td>(m,</td><td colspan="2">9H), 6.56 (</td><td>: dd,</td><td>J = 9.1,</td><td>1.7 Hz,</td><td>1 HOUR) ,</td><td> 5.03</td><td>(dd,</td><td>J =</td>

<td> 10.6,</td><td> 9.3</td><td>Hz</td><td>, 1H), 4</td><td> . 73</td><td>(s, 1H),</td><td>4.05 (dd,</td><td>J =</td><td> 11.5,</td><td> 3.3</td><td>Hz,</td>

<td>1 HOUR) ,</td><td> 3.51</td><td> -</td><td>3.37 (m,</td><td>2H)</td><td> , 3.25 -</td><td>3.15 (m,</td><td>2H),</td><td> 3.09</td><td>(s,</td><td>3H),</td>

<td> 2.92</td><td>(td,</td><td>J</td><td> = 12.5,</td><td> 2.9</td><td>Hz, 1H),</td><td>2.31 (t,</td><td>J =</td><td> 7.2</td><td>Hz,</td><td>2H),</td>

<td> 2.18</td><td> - 1.</td><td> 70</td><td>(m, 15H)</td><td> , 1.</td><td>61 (ddt,</td><td>J = 12.8,</td><td> 8.4,</td><td> 4.9</td><td>Hz,</td><td>1 HOUR) ,</td>

<td> 1.28</td><td>(dd,</td><td>J</td><td> = 30.1,</td><td> 6.7</td><td>Hz, 7H),</td><td>1.04 (s,</td><td>9H),</td><td> 0.88</td><td>(dd,</td><td>J =</td>

<td> 37.3,</td><td> 6.5</td><td>Hz</td><td>, 6H).</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

Finally, the trifyl protected thiol dissolved in

275

CH₂C1₂ (3 ml) and trifluoroacetic acid (0.6 ml) with triisopropyl silane (0.1 ml) was added. The reaction was monitored by HPLC-MS and upon completion, it was concentrated to dryness under reduced pressure. The residue was taken up in CH₂C1₂ (~ 0.8 mi) with a couple of drops of ethanol and cooled to 0^or C in an ice bath. The cold diethyl ether (~ 3 mL) was added with vigorous stirring to generate a white precipitate that was collected by filtration in a Buchner funnel dried under high vacuum to give the original compound as an amorphous white solid.

^XH NMR (400 MHz, Methanol-d₄) δ 6.52 (d, J = 9.0 Hz, 1H), 5.06

<td>(dd,</td><td>J = 10.7, 8.8</td><td>Hz, 1H</td><td> ) , 4.73 (</td><td>s, 1H), 4.16 - 4.04</td><td>(m,</td>

<td>1 HOUR) ,</td><td>3.69 - 3.56 (m,</td><td>• 2H),</td><td>3.48 (dd,</td><td>J = 13.3, 7.2 Hz,</td><td>2H),</td>

<td> 3.15</td><td>(s, 3H), 3.03 -</td><td> 2.94 (</td><td>m, 1H), 2</td><td>.68 (t, J = 6.9 Hz,</td><td>1 HOUR) ,</td>

<td> 2.24</td><td>- 1.77 (m, 11H)</td><td> , 1.61</td><td>(s, 1H),</td><td>1.31 (dd, J = 27.2,</td><td> 6.7</td>

<td>Hz,</td><td>6H), 1.06 (s, 9H)</td><td> , 0.91</td><td>(dd, J =</td><td>34.1, 6.6 Hz, 6H).</td><td></td>

Example 116

<img file="MX368258B_D0174.tif" />

Chemical Formula: C₂₉H₄₆N₄OR_sS₂

Exact Mass: 594.29 (116)

S) -N - ((S) -1 - ((S) -2 - ((E) -3- (3-mercaptopropylsulfonamido) -2276 methyl-3-oxoprop-l-enyl) pyrrolidin-l-yl) - 3,3-dimethyl-loxobutan-2-yl) -3-methyl-2- (methylamino) -3-phenylbutanamide

The title compound was synthesized from Boc-proline and the Example according to General Procedures 10,

11, 2, 3, 7 and others from Nieman JA et al. J.

Prod. 2003,

66, 183-199. The compound was isolated as two diastereoisomers in a ratio of approximately

1:1.

<td colspan="3">NMR (400 MHz, Methanol-d₄)</td><td colspan="2">δ 7.57 - 7.12</td><td colspan="3">(m, 5H), 6.39</td><td>(dd,</td>

<td>J =</td><td>9.4, 1.6 Hz,</td><td>0.5H), 6.2</td><td>51 (dd, J =</td><td> : 8</td><td> .2, 1.5</td><td>Hz,</td><td> 0.</td><td>5H),</td>

<td> 4.72</td><td>(q, J = 7.5</td><td>Hz, 0.5H),</td><td colspan="2"> 4.66 - 4.56 (</td><td>m, 0.5H</td><td> ), 4</td><td> .40</td><td>(s,</td>

<td>0.5H</td><td>), 4.28 (d, J</td><td>= 11.9 Hz,</td><td>1H), 3.81 I</td><td>: m,</td><td>0.5H),</td><td> 3.76</td><td> -</td><td> 3.56</td>

<td>(m,</td><td>3H), 2.77-2</td><td>.64 (m, 2H)</td><td>, 2.59 (m,</td><td>3H</td><td> ), 2.39</td><td> - 2</td><td> .22</td><td>(m,</td>

<td>1 HOUR) ,</td><td> 2.18 - 1.72</td><td>(m, 7H), 1</td><td> .61 - 1.33</td><td>(m,</td><td>- 6H),:</td><td> 1.15</td><td> —</td><td> 0.85</td>

11H).

C29H46N4O5S2 caled m / z = 594.35 found [M + H]

595.3

Example 117

<img file="MX368258B_D0175.tif" />

<img file="MX368258B_D0176.tif" />

(117) (S) -N- ((S) -1- (2- (3- (3-mercaptopropylsulfonamido) -2-methyl-3 oxoprop-l-enyl) piperidin-l-yl) -3,3- dimethyl-l-oxobutan-2-yl) 277

3-methyl-2- (methylamino) -3-phenylbutanamide

The title compound was synthesized from Bochomoproline and Example 2 according to General Procedures 10, 11, 2, 3, 7 and others from Nieman JA et al. J. Nat. Prod. 2003, 66, 183-199. The compound was isolated as two diastereoisomers in a ratio of approximately 2: 3.

^ΧΗ NMR (600 MHz, Methanol-d₄) δ 7.55 (d, J = 7.8 Hz, 1H), 7.46 (m, 3H), 7.38 (m, 1H), 6.81 (d, J = 8.3 Hz, 0.6H), 6.79 (d, J = 7.8 Hz, 0.4H), 5.66 (m, 0.6H), 5.12 (m, 0.4H), 5.05 (s,

0.6H), 4.86 (s, 0.4H), 4.42 (d, J = 14.9 Hz, 0.4H), 4.35 (s,

0.6H), 4.26 (s, 0.4H), 4.12 (d, J = 13.8 Hz, 0.6H), 3.64 (d,

J = 7.6 Hz, 1H), 3.63 (d, J = 7.4 Hz, 1H), 3.39 (m, 0.6H),

2.94 (td, J = 13.8, 2.6 Hz, 0.4H), 2.68 (t, J = 6.7 Hz, 2H), 2.56 (m, 3H), 2.10 (m, 3.5H), 1.97 (s, 1.5H), 1.90-1.70 (m, 7H), 1.65-1.29 (m, 6H), 1.07 (s, 3.5H), 1.04 (s, 4.5H) ppm. C₃oH₄7N₄0₅S₂ caled, m / z = 608.31; found [M + H]⁺ = 609.32

Example 118

<img file="MX368258B_D0177.tif" />

(118)

278 (S) -N- ((S) -1- (2- (3- (4- (mercaptomethyl) phenylsulfonamido) -2methyl-3-oxoprop-l-enyl) piperidin-l-yl) -3,3-dimethyl -loxobutan-2-yl) -3-methyl-2- (methylamino) -3-phenylbutanamide

The title compound was synthesized from Bochomoproline and Example 7 according to General Procedures 10, 11, 2, 3, 7 and others from Nieman JA et al. J. Nat. Prod. 2003, 66, 183-199. The compound was isolated as two diastereoisomers in a ratio of approximately 2: 3.

¹H NMR (600 MHz, Methanol-d₄) δ 8.02 (d, J = 8.4 Hz, 0.8H), 8.00 (d, J = 8.5 Hz, 1.2H), 7.58 (d, J = 8.5 Hz, 1H), 7.54

<td>(d, J = 8.5</td><td>Hz, 2H),</td><td>7.45 (t, J =</td><td>8.2 Hz</td><td colspan="2">, 2H), 7.40 (d,</td><td>J =</td>

<td>7.2 Hz, 0.6H)</td><td> , 7.36</td><td>(m, 1H), 7.31 (</td><td>t, J =</td><td> 7.1</td><td>Hz, 0.4H), 6</td><td> .74</td>

<td colspan="2">(d, J = 8.2 Hz, 1H),</td><td>5.59 (m, 0.6H)</td><td> , 5.06</td><td>(m,</td><td>0.4H), 5.02</td><td>(s,</td>

<td>0.6H), 4.84</td><td>(s, 0.4H</td><td>), 4.39 (d, J </td><td> = 12.5</td><td>Hz,</td><td>0.4H), 4.34</td><td>(s,</td>

<td>0.6H), 4.20</td><td>(s, 0.4H:</td><td>), 4.08 (d, J </td><td> = 12.0</td><td>Hz,</td><td>0.6H), 3.83</td><td>(s,</td>

<td>1.2H), 3.73</td><td colspan="2">(s, 0.8H), 3.35 (m, 0</td><td>.6H), 2</td><td> . 93</td><td>(td, J = 13</td><td>• 6 t</td>

<td>3.0 Hz, 0.4H)</td><td> , 2.55 1</td><td>(m, 3H), 2.00 (</td><td>s, 1H),</td><td> 1.</td><td colspan="2">90-1.51 (m, 7H),</td>

<td>1.51-1.30 (m</td><td>, 4H),</td><td>1.30 (s, 1H),</td><td> 1.15</td><td>(s,</td><td>1H), 1.04</td><td>(s,</td>

<td>3.5H), 1.01 (</td><td>s, 4.5H)</td><td>ppm.</td><td></td><td></td><td></td><td></td>

C34H47N4O5S2 caled, m / z = 656.31; found [M + H]

657.30

<img file="MX368258B_D0178.tif" />

279

Example 119

OR

<img file="MX368258B_D0179.tif" />

MC-VC-PABC-77

The title compound was prepared by the application of general procedures 15 and 7 from Example 77 protected with Boc.

^XH NMR (400 MHz, Methanol-d₄) δ 7.58 (d, J = 8.2 Hz, 2H), 7.49

<td>(d,</td><td colspan="3">J = 7.5 Hz, 2H),</td><td> 7.38</td><td>(t, J</td><td colspan="2"> = 7.7</td><td colspan="2">Hz, 2H), 7.36 -</td><td> 7.24</td>

<td>(m,</td><td>6H), 7.22</td><td>(d, J</td><td> =</td><td colspan="2">7.8 Hz, 2H)</td><td> , 6.</td><td> 81</td><td>(s, 2H)</td><td>, 6.57 (d,</td><td>J =</td>

<td> 9.1</td><td>Hz, 1H),</td><td> 5.08 (</td><td>s,</td><td>2H),</td><td> 5.04</td><td>(t,</td><td>J</td><td> = 10.0</td><td>Hz, 1H),</td><td> 4.91</td>

<td>(s,</td><td>1H), 4.53</td><td>(dd,</td><td>J</td><td> = 9.0,</td><td> 5.1</td><td>Hz,</td><td>1 HOUR)</td><td> , 4.40</td><td>(s, 2H),</td><td> 4.28</td>

<td>(s,</td><td>2H), 4.19</td><td>(d, J</td><td> =</td><td colspan="2">7.4 Hz, 1H)</td><td> , 3,</td><td> .49</td><td>(t, J -</td><td>= 7.1 Hz,</td><td>2H),</td>

3.26 - 3.11 (m, 2H), 3.07 - 2.93 (m, 3H), 2.30 (t, J = 7.4

Hz, 2H), 2.18 (s, 3H), 2.15 - 2.05 (m, 1H), 1.99 - 1.91 (m,

<td colspan="2">1H), 1.89</td><td colspan="2">(s, 3H), 1.83 - 1.72 (m,</td><td>1H), 1.72 -</td><td> 1.53</td><td>(m, 7H),</td>

<td> 1.44</td><td>(s,</td><td>3H), 1.37 (</td><td>s, 3H), 1.35</td><td>- 1.27 (m,</td><td>2H),</td><td>1.03 (s,</td>

<td>9H),</td><td> 1.00</td><td>(d, J = 6.8</td><td>Hz, 3H), 0.99</td><td>(d, J = 6.7</td><td>Hz,</td><td>3H), 0.88</td>

(d, J = 6.5 Hz, 3H), 0.82 (d, J = 6.6 Hz, 3H).

280

C64H91N11O13S caled, m / z = 1253.7; found [M + H]⁺ = 1254.8.

Example 120

<img file="MX368258B_D0180.tif" />

(120)

4 - ((R) -2 - ((R) -2- (6- (2,5-dioxo-2,5-dihydro-lH-pyrrol-lyl) hexanamido) -3-methylbutanamide) -5-ureidopentanamide) benzyl

4- (N - ((S, E) -2,5-dimethyl-4 - ((S) -N, 3,3-trimethyl-2 - ((S) -3-methyl2- (methylamino) -3- phenylbutanamido) butanamido) hex-2enoyl) sulfamoyl) benzylcarbamate,

MC-VC-PABC-85

The title compound was prepared by the application of general procedures 15 and 7 to Example 85 protected with Boc.

CesHggNnOisS caled, m / z = 1239.6; found [M + H]⁺ = 1240.9.

281

Example 121

<img file="MX368258B_D0181.tif" />

(121)

MC-VC-PABC-80

The title compound was prepared by the application of general procedures 15 and 7 to Example 80 protected with Boc.

C53H89N11O13S caled, m / z = 1239.6; found [M + H]⁺ = 1240.9.

Example 122

<img file="MX368258B_D0182.tif" />

MC-VC-PABC-41

282

The title compound was prepared by the application of General Procedure 15 to Example 41.

C₆4H₉iNuOi3S caled, m / z = 1253.65; found [M + H]⁺ = 1254.75, [M + 2H] ²⁺ = 628.20.

Example 123

<img file="MX368258B_D0183.tif" />

Chemical formula:

Exact Mass: 628.37

Molecular Weight: 628 87 (123) (R) -N- (benzylsulfonyl) -2,5-dimethyl-4 - ((S) -N, 3,3-trimethyl-2 ((S) -3-methyl-2 - (methylamino) -3phenylbutanamide) butanamide) hexanamide

A suspension of Example 14 and 10% palladium on carbon (25 mol% Pd) in glacial acetic acid was stirred under an atmosphere of H₂ (1 atm) at room temperature. After 142 h, the reaction suspension was passed through a celite bed, rinsed with MeOH (5x) and concentrated in vacuo. The light brown residual crude was dissolved and purified in preparative HPLC (30-70% MeCN / H₂0 with 0.1% TEA) and lyophilized to give a reduced product diastereomer as a pale yellow solid in a yield of

283

15%.

¹H NMR (400 MHz, Methanol-d₄) δ 7.55 (d, J = 7.2 Hz, 2H), 7.46 (t, J = 7.8 Hz, 2H), 7.43 - 7.31 (m, 6H), 5.01 (s, 1H), 4.79 (d, J = 14.1 Hz , 1H), 4.65 (d, J = 14.1 Hz, 1H), 4.35 (s, 1H), 4.24 (s, 1H), 3.07 (s, 3H), 2.52 (s, 3H), 2.27 (m, J = 10.3, 7.0, 3.2 Hz, 1H), 2.14 (ddd, J = 13.5, 10.6, 2.7 Hz, 2H), 1.78 (d, J = 8.6 Hz, 1H), 1.47 (s, 3H), 1.34 (s, 3H ), 1.15 (d, J = 6.9 Hz, 3H), 1.14 (s, 9H), 1.04 (d, J = 6.6 Hz, 3H), 0.82 (d, J = 6.6 Hz, 3H).

C3₄H₅₂N₄OR₅S caled m / z = 628.37 amu; found [M + H]⁺ = 629.6, [M + Na]⁺ = 651.6

GENERAL SYNTHESIS SCHEMES FOR (T) - (L) - (D)

USE OF LC-SPDP and SMCC LINKERS

<img file="MX368258B_D0184.tif" />

mAb-SPDP mAb mAb-SPDP-SR₂'-peptide-NHS0₂Ri

Composition produced using the SPDP binding method described below.

Note that R₂'is different from R₂, since R₂ includes R₂'-S.

284

<img file="MX368258B_D0185.tif" />

<img file="MX368258B_D0186.tif" />

mAb-SMCC-SR₂'-peptide-NHSO₂Ri

Composition produced using the SMCC binding method described below. Note that R₂'is different from R₂, since R₂ includes R₂'-S

<img file="MX368258B_D0187.tif" />

peptide-NHS0₂Ri'-S-SPDP

Composition produced using the SPDP binding method described below.

285

<img file="MX368258B_D0188.tif" />

<img file="MX368258B_D0189.tif" />

peptide-NHS0₂Ri '-S-SMCC

Composition produced using the SMCC binding method described below. Note that R / is different from Ri, since Ri includes R / -S

Example 124

<img file="MX368258B_D0190.tif" />

(124) (Compound A-SPDP-mAb) produced using the synthesis method of Compound A, above, and the SPDP binding method described below.

286

Example 125

<img file="MX368258B_D0191.tif" />

(125) (Compound B - SPDP-mAb) produced using the synthesis method of Compound B, above, and the SPDP binding method described below

Example 126

<img file="MX368258B_D0192.tif" />

Οχ / O

N '^S'H

<img file="MX368258B_D0193.tif" />

^ss

H ^N((126) (Compound C-SPDP-mAb) produced using the synthesis method of Compound C, above, and the SPDP binding method described below.

Example 127

<img file="MX368258B_D0194.tif" />

mAb (Compound B - SMCC - mAb) produced using the method of

287 Synthesis of Compound B, above, and SMCC link described below.

Example 128

<img file="MX368258B_D0195.tif" />

(Compound A - SMCC-mAb) synthesis of Compound A SMCC link described to the method

<img file="MX368258B_D0196.tif" />

mAb produced using previously, and then.

the method method

Example 129

<img file="MX368258B_D0197.tif" />

mAb (Compound C-SMCC-mAb) produced using the synthesis method of Compound C, above, and the SMCC binding method described below.

Example 130

<img file="MX368258B_D0198.tif" />

(130)

288

3-methyl-3- (4-bromophenyl) -butanic acid

To a vigorously stirred solution of bromobenzene (4.70 g, 30.0 mmol) and 3,3-dimethylacrylic acid (1.00 g, 10.0 mmol) in 20 ml of CH₂C1₂ cooled to -10 ° C in an NH bath₄Cl_(here) / dry ice, solid AICI3 was added in portions, keeping the internal temperature below -5 ° C. The solution turned yellow, then brown after the addition. After one hour, the analysis by LC and CCF indicated the complete consumption of the limiting reagent. Then, the reaction was quenched by the addition of 1M citric acid causing the brown color to fade to yellow. The resulting semi-solid suspension was extracted four times with 20 ml of Et₂Or, the combined organic extracts were washed with NaCl. _(sat), dried over Na₂SW_{4 (s)}, and concentrated in vacuo with heating at 45 ° C to remove solvent and residual bromobenzene. The resulting oil solidified slowly. Recrystallization of the crude solid in hexanes provided the title compound (1.29 g, 50%) as clusters of white prisms.

¹H NMR (400 MHz, Chlorof ormo-d) δ (ppm) 7.42 (d, J = 8.6 Hz, 2H), 7.23 (d, J = 8.6 Hz, 2H), 2.63 (s, 2H), 1.43 (s, 6H). CiiHi3BrO2 caled. [M + H]⁺ = 257.02 amu; found m / z = 257.03. Rf = 0.21 (20% (2% AcOH / EtOAc) / Hex).

289

Example 131

<img file="MX368258B_D0199.tif" />

Br (131)

3-methyl-3- (3-bromophenyl) -butanic acid

The title compound was prepared in the same manner as 3-methyl-3-phenylbutanoic acid in Nieman JA, et al. J. Nat Prod. 2003, 66, 183-199, using bromobenzene instead of benzene as a solvent, and replacing the acid-base work with a simple extraction of the 1M citric acid reaction mixture and three successive recrystallizations of hexanes. From a crude product enriched in the desired target isomer as a 2: 1 mixture, the title compound could be obtained in the form of white short needles in more than 95% purity.

^ΧΗ NMR (400 MHz, Chloroform-d) δ (ppm) 7.49 (t, J = 1.9 Hz, 1H), 7.34 (ddd, J = 7.9, 1.9, 1.0 Hz, 1H), 7.29 (ddd, J = 7.9 1.9 , 1.0 Hz, 1H), 7.18 (t, J = 7.9 Hz, 1H), 2.64 (s, 2H), 1.44 (s, 6H). CnHi₃BrO₂ caled [M + H]⁺ = 257.02 amu; found m / z = 257.01. R_F = 0.21 (20% (2% AcOH / EtOAc) / Hex).

290

Example 132

Br

OR

<img file="MX368258B_D0200.tif" />

OMe (132) (S) -methyl 3- (4-bromophenyl) -2- (tert butoxycarbonyl (methyl) amino) -3-methylbutanoate

The title compound is synthesized according to Example 130 according to the sequence of procedures described by Nieman et al. for the synthesis of (S) -methyl 2- (tertbutoxycarbonyl (methyl) amino) -3-methyl-3-phenylbutanoate.

Example 133

OH _zNBoc (133)

(S) -2 - ((tert-butoxycarbonyl) (methyl) amino) -3- (4 - ((14-hydroxy-3,6,9,12-tetraoxatetradecyl) oxy) phenyl) -3-methylbutanic acid

To a stirred solution of Example 68 (157 mg, 0.405 mmol) in pentaethylene glycol (1.5 ml) CsCO was added₃ (330 mg, 1.01 mmol), 3,4,7,8-tetramethyl-l, 10-phenanthroline (57 mg, 0.24 mmol), and Cul (23 mg, 0.12 mmol). It was gasified with nitrogen in

291 The flask was then sealed and heated to 130 ° C, the solution quickly turned red to brown to black. After 40 h, the reaction appeared to be almost complete by HPLC analysis. Therefore, the mixture was allowed to cool to room temperature, diluted with H₂Or, and transferred to a larger Erlenmeyer with a stir bar. This mixture was carefully acidified to pH ~ 3 with 1M citric acid, paying attention not to allow the foamy mixture to spill. Then, the mixture was extracted five times with CH₂C1₂, the combined organic extracts were washed with NaCl (sat) r dried over Na₂SW_{4 (s)}, and concentrated in vacuo to give approximately 300 mg of crude oil. Purification by flash chromatography (1-10% MeOH / (2% AcOH / EtOAc)) produced the title compound (66 mg, 30%) as a clear film that existed as a set of NBoc rotamers and a ratio of approximately 2: 1.

¹H NMR (400 MHz, Chloroform-d) δ (ppm) 7.35 (d, J = 7.8 Hz, 1.3H), 7.30 (d, J = 7.6 Hz, 0.7H), 6.87 (d, J = 7.1 Hz, 2H ), 5.07 (s, 0.7H), 4.93 (s, 0.3H), 4.14 (m, 2H), 3.86 (m, 2H), 3.70 (m, 16H), 2.83 (s, 1H), 2.72 (s, 2H), 1.54 (s, 3H), 1.49 (s, 3H), 1.45 (s, 9H). C₂7H₄₅DO NOT₁₀ caled [M + H]⁺ = 544.31 amu; found m / z = 544.36. Rf = 0.36 (5% MeOH / (2% AcOH / EtOAc)).

292

HO

Example 134

OR

<img file="MX368258B_D0201.tif" />

ΌΗ (134)

(S) -2 - ((tert-butoxycarbonyl) (methyl) amino) -3- (4- (2- (2 (2- (2-hydroxyethoxy) ethoxy) ethoxy) ethoxy) phenyl) -3-methylbutanic acid The title compound was prepared according to the previous method of Example 68 (132 mg, 0.341 mmol), CsCO3 (278 mg, 0.853 mmol), 3,4,7,8-tetramethyl-l, 10-phenanthroline (24 mg, 0.10 mmol), and Cul (10 mg, 0.051 mmol). Flash chromatography (1-10% MeOH / (2% AcOH / EtOAc)) gave the title compound (66 mg, 38%) as a clear oil with an approximate 2: 1 ratio of N-Boc rotamers.

¹H NMR (400 MHz, Chloroform-d) δ (ppm) 7.34 (d, J = 8.4 Hz, 1.3H), 7.29 (d, J = 8.1 Hz, 0.7H), 6.85 (d, J = 8.4 Hz, 2H ), 5.05 (s, 0.7H), 4.91 (s, 0.3H), 4.13 (t, J = 4.6 Hz, 2H), 3.87 - 3.79 (m, 2H), 3.76 - 3.60 (m, 10H), 3.59 ( t, J = 4.1 Hz, 2H), 2.80 (s, 1H), 2.69 (s, 2H), 1.53 (s, 3H), 1.48 (s, 3H), 1.44 (s, 9H). C2₅H₄₁DO NOT₉ caled [M + H]⁺ = 500.29 amu;

found m / z = 500.36. R_F = 0.46 (5% MeOH / (2% AcOH / EtOAc)).

293

Example 135

<img file="MX368258B_D0202.tif" />

(135)

(5) -3- (3 - ((14-hydroxy-3,6,9,12 tetraoxatetradecyl) oxy) phenyl) -3-methyl-2- (methylamino) butanic acid The precursor of the title compound, acid (5) -3- (3-bromophenyl) -2 - ((tert-butoxycarbonyl) (methyl) amino) -3-methylbutanic, was prepared from Example 131 following the procedures of Neiman et al.

Therefore, following the above procedures, from (5) -3- (3-bromophenyl) -2 - ((tertbutoxycarbonyl) (methyl) amino) -3-methylbutanic acid (166 mg, 0.43 mmol), CsCCb (330 mg, 1.01 mmol), 3.4, Ί, 8-tetramethyl-l, 10 phenanthroline (31 mg, 0.13 mmol), and Cul (12.3, 0.060 mmol) in 1.5 mL of pentaethylene glycol heated at 130 ° C for two days, obtained the title compound (73 mg, 31%) as a clear oil after flash chromatography (1-10% MeOH / (2% AcOH / EtOAc)) in an approximately 2: 1 ratio of N-Boc rotamers.

¹H NMR (400 MHz, Chloroform-d) δ (ppm) 7.17 (t, J - 7.8 Hz,

1H), 7.14-7.07 (m, 1H), 7.07-6.93 (m, 2H), 6.74 (d, J

294

8.0 Hz, 1H), 5.11 (s, 0.7H), 4.93 (s, 0.3H), 4.25 - 4.03 (m,

2H), 3.91 - 3.77 (m, 2H), 3.78 - 3.66 (m, 2H), 3.69 - 3.43 (s,

14H), 2.72 (s, 1H), 2.65 (s, 1H), 1.51 (s, 3H), 1.49 (s, 3H),

1.45 (s, 9H). C27H45NO10 caled. [M + H]⁺ = 544.31 amu; found m / z = 544.34.

Example 136

AcS (136) (6S, 9S, 12S, E) -ethyl 9- (tert-butyl) -12-isopropyl-2,2,5,11,14pentamethyl-4,7,10-trioxo-6- (2- (4 - ((16-oxo-3,6,9,12-tetraoxa15-thiaheptadecyl) oxy) phenyl) propan-2-yl) -3-oxa-5,8,11triazapentadec-13-en-15-oato

(S) -2 - ((tert-butoxycarbonyl) (methyl) amino) -3- (4 - ((14-hydroxy-3,6,9,12-tetraoxatetradecyl) oxy) phenyl) -3-methylbutanic acid (65 mg, 0.120 mmol) was coupled to (S, E) -ethyl 4 ((S) -2-amino-N, 3,3-trimethylbutanamido) -2,5-dimethylhex-2enoate with HATU and DIPEA following the same stoichiometry and procedure as described in the general coupling procedures in Nieman et al. to give a free intermediate alcohol after purification by flash chromatography (1-10% MeOH / (2% AcOH / EtOAc)). TO

295 then to triphenylphosphine (40 mg, 0.15 inmol) in THF 0.75 ml under N₂ to 0^or C, di-tertbutylazodicarboxylate (35 mg, 0.15 mmol) was added in one portion. After 35 minutes, a white precipitate crashed and the reaction became difficult to stir. To this suspension, a solution of the intermediate alcohol (42 mg, 0.050 mmol) in 0.75 ml of THF was added diluting the precipitate sufficiently to restore the stirring. Five minutes later, thioacetic acid (5.7 mg, 0.075 mmol) in 0.05 ml of THF was added causing all the yellow color to disappear from the mixture. After 30 min, the reaction was allowed to warm to room temperature. The precipitate disappeared after another 15 min, and the analysis by CCF and LCMS showed an almost complete conversion. After another 40 minutes, the reaction mixture was concentrated in vacuo, then subjected directly to flash chromatography (40-100% EtOAc / Hex then to 10% MeOH / EtOAc) to give the title compound (26 mg, 57% ) as a clear movie.

¹H NMR (400 MHz, Chloroform-d) δ (ppm) 7.43 (d, J = 8.4 Hz, 1.3H), 7.31 (d, J = 8.3 Hz, 0.7H), 6.97 - 6.72 (m, 2H), 6.62 (dd, J = 9.3, 1.6 Hz, 1H), 6.14 (d, J = 9.6 Hz, 1H), 5.22 (s, 0.7H), 5.12 - 4.99 (m, 1H), 4.84 (s, 0.3H), 4.69 (d, J = 9.3 Hz, 0.3H), 4.60 (d, J = 8.9 Hz, 0.7H), 4.19 (q, J = 7.2 Hz,

296

2Η), 4.09 (td, J = 4.6, 2.3 Hz, 2H), 3.84 (t, J = 4.9 Hz, 2H),

3.77 - 3.70 (m, 2H), 3.70 - 3.61 (m, 10H), 3.59 (t, J = 6.4 Hz, 2H), 3.07 (t, J = 6.4 Hz, 2H), 2.97 - 2.91 (m, 3H) , 2.84 (s, 3H), 2.32 (s, 3H), 1.87 (s, 3H), 1.49 (s, 3H), 1.43 (s, 9H), 1.35 (s, 3H), 1.30 (t, J = 7.1 Hz, 3H), 0.87 (d, J = 6.6 Hz, 3H), 0.80 (d, &lt;J = 16.6 Hz, 3H), 0.77 (s, 9H). C46H77N3O12S caled. [M + H]⁺ = 896.53 amu; found m / z = 896.77. R_F = 0.56 (80% EtOAc / Hex).

Example 137

<img file="MX368258B_D0203.tif" />

(137) (6S, 90,125.5) -ethyl 9- (tert-butyl) -12-isopropyl-2,2,5,11,14pentamethyl-4,7,10-trioxo-6- (2- (4- ((13-oxo-3, 6, 9-trioxa-12tiatetradecyl) oxy) phenyl) propan-2-yl) -3-oxa-5,8,11 triazapentadec-13-en-15-oato

The title compound was prepared from (5) -2 ((tert-butoxycarbonyl) (methyl) amino) -3- (4- (2- (2- (2- (2 (hydroxyethoxy) ethoxy) ethoxy)) ethoxy)) phenyl) -3-methylbutanic (66 mg, 0.065 mmol) following the same procedure described above to give 32 mg (57%) as a clear film after flash chromatography (20-100% EtOAc / Hex).

297 '• H NMR (400 MHz, Chloroform-d) δ (ppm) 7.44 (d, J = 8.5 Hz, 1.3H), 7.32 (d, J = 8.5 Hz, 0.7H), 6.95 - 6.77 (m, 2H) , 6.62 (dd, J = 9.2, 1.7 Hz, 1H), 6.09 (d, J = 9.1 Hz, 1H), 5.24 (s, 0.7H), 5.13 - 4.95 (m, 1H), 4.84 (s, 0.3H ), 4.69 (d, J = 9.6 Hz, 0.3H), 4.60 (d, J = 9.0 Hz, 0.7H), 4.19 (q, J = 7.1 Hz,

2H), 4.09 (td, J = 4.7, 2.4 Hz, 2H), 3.84 (t, J = 4.9 Hz, 2H)

3.72 (dd, J = 5.7, 3.2 Hz, 2H), 3.70 - 3.65 (m, 2H), 3.66 3.62 (m, 4H), 3.60 (t, J = 6.5 Hz, 2H), 3.09 (t, J = 6.5 Hz,

2H), 2.96 - 2.88 (m, 3H), 2.84 (s, 3H), 2.33 (s, 3H), 1.88 (d J = 3.5 Hz, 3H), 1.49 (s, 2H), 1.43 (d, J = 5.5 Hz, 11H), 1.35 (s, 2H), 1.30 (t, J = 7.1 Hz, 2H), 0.87 (d, J = 6.6 Hz, 3H), 0.80 (d, J = 15.9 Hz, 3H), 0.76 (s, 9H). C₄₄H₇₃N₃0nS caled. [M + H]⁺ = 852.51 amu; found m / z = 852.79. R_F = 0.60 (60% EtOAc / Hex).

Example 138

AcS (138) (6S, 9S, 12S, E) -ethyl 9- (tert-butyl) -12-isopropyl-2,2,5,11,14pentamethyl-4,7,10-trioxo-6- (2- (3 - ((16-oxo-3,6,9-trioxa-12 tiatetradecyl) oxy) phenyl) propan-2-yl) -3-oxa-5, 8,11298 triazapentadec-13-en-15-oato

The title compound was prepared from (S) -3 (3 - ((14-hydroxy-3,6,9,12-tetraoxatetradecyl) oxy) phenyl) -3-methyl-2- (methylamino) butanic acid (73 mg , 0.080 mmol) following the same procedure described above to give 66 mg (47%) as a clear film after flash chromatography (20-100% EtOAc / Hex).

^XH NMR (400 MHz, Chloroform-d) δ (ppm) 7.25 - 6.92 (m, 3H), 6.78 - 6.70 (m, 1H), 6.62 (d, J = 8.9 Hz, 1H), 6.12 (d, J = 8.9 Hz, 1H), 5.26 (s, 0.7H), 5.12 - 4.99 (m, 1H), 4.89 (s, 0.3H), 4.74 - 4.56 (m, 1H), 4.19 (q, J = 1.2 Hz, 1H ), 4.16 4.03 (m, 2H), 3.84 (td, J = 5.0, 3.2 Hz, 2H), 3.77 - 3.61 (m, 14H), 3.60 (t, J = 6.4 Hz, 2H), 3.09 (t, J = 6.5 Hz, 2H), 2.97 - 2.75 (m, 6H), 2.33 (s, 3H), 1.91 - 1.83 (m, 3H), 1.52 - 1.35 (m, 16H), 1.26 (t, J = 7.1 Hz, 3H), 0.87 (d, J = 6.0 Hz, 3H), 0.81 (d, J = 12.9 Hz, 3H), 0.77 (s, 9H). C46H77N3O12S caled. [M + H]⁺ = 896.53 amu; found m / z = 896.68. R_F = 0.61 (75% EtOAc / Hex).

Example 139 (139)

299

Acid disulfuric acid (S, E) -4 - ((5) -2 - ((S) -3- (4 - ((14-mercapto-

3,6,9,12-tetraoxatetradecyl) oxy) phenyl) -3-methyl-2 (methylamino) butanamido) -N, 3,3-trimethylbutanamido) -2,5-dimethylhex-2-oenic

The title compound was prepared by saponification, subsequently the TFA promotes the elimination of Be, according to the exact methods described in Nieman et al. from (65.95,125, E) -ethyl 9- (tert-butyl) -12-isopropyl-

2,2,5,11,14-pentamethyl-4,7,10-trioxo-6- (2- (4 - ((16-oxo-

3,6,9,12-tetraoxa-15-thiaheptadecyl) oxy) phenyl) propan-2-yl) -3oxa-5,8, ll-triazapentadec-13-en-15-oato (26 mg, 0.029 mmol) for give the title compound (16 mg, 90%) as a clear glass after complete removal of excess TFA.

NMR (400 MHz, Methanol-d₄) δ (ppm) 8.43 (d, J = 8.1 Hz, 1H),

7.47 (d, J = 8.5 Hz, 2H), 7.08 - 6.94 (m, 2H), 6.80 (dq, J =

9.9, 1.5 Hz, 1H), 5.08 (t, J = 10.1 Hz, 1H), 4.94 (d, J = 8.1 Hz, 1H), 4.32 (s, 1H), 4.21 - 4.12 (m, 2H), 3.93 - 3.81 (m,

3H), 3.76 (t, J = 6.4 Hz, 2H), 3.76 - 3.72 (m, 2H), 3.72 3.62 (m, 10H), 3.17 (s, 3H), 2.92 (t, J = 6.4 Hz, 2H) , 2.61 -

2.47 (m, 3H), 2.14 - 2.00 (m, 1H), 1.94 (d, J = 1.5 Hz, 3H),

1.46 (s, 3H), 1.40 (d, J = 7.7 Hz, 3H), 1.09 (s, 9H), 0.94 (d

J = 5.0 Hz, 3H), 0.92 (d, J = 4.8 Hz, 3H). C₇4Hi2₄N₆OR₁₈S2 caled. [M + H]⁺ = 1449.85 amu; found m / z = 1450.49.

300

HS

Example 140

OR

OR

<img file="MX368258B_D0204.tif" />

ΌΗ (140)

The compound of Example 139 is reduced according to the methods below to produce the compound in question.

Example 141 (141)

Acid disulfide (S, E) -4 - ((S) -2 - {(S) -3- (4- (2- (2- (2- (2mercaptoethoxy) ethoxy) ethoxy) ethoxy) phenyl) -3 -methyl-2 (methylamino) butanamido) -N, 3,3-trimethylbutanamido) -2,5 dimethylhex-2-enóico

The title compound was prepared by saponification, subsequently the TFA promoted the elimination of Boc, according to the exact methods described in Nieman et al. from (6S, 9S, 12S, E) -ethyl 9- (tert-butyl) -12-isopropyl-

2,2,5,11,14-pentamethyl-4,7,10-trioxo-6- (2- (4 - ((13-oxo-3,6,9 trioxa-12-tiatetradecyl) oxy) phenyl) propan -2-yl) -3-oxa-5,8,11

301 triazapentadec-13-en-15-oato (32 mg, 0.037 mmol) to give the title compound (29 mg, 86%) as a clear glass after complete removal of excess TFA.

NMR (400 MHz, Methanol-d₄) δ (ppm) 8.39 (d, J = 8.2 Hz, 1H), 7.44 (d, 8.9 Hz, 2H), 7.01 (d, 8.5 Hz, 2H), 6.77 (d,

J = 7.9 Hz, 1H), 5.05 (t, J = 10.1 Hz, 1H), 4.92 (d, J = 8.3

Hz, 1H), 4.28 (s, 1H), 4.15 (dd, J = 5.8, 3.4 Hz, 2H), 3.89 3.80 (m, 2H), 3.73 (t, J = 6.4 Hz, 2H), 3.72 - 3.69 ( m, 2H),

3.69 - 3.60 (m, 6H), 3.14 (s, 3H), 2.89 (t, J = 6.4 Hz, 2H),

2.50 (s, 3H), 2.11 - 1.97 (m, 1H), 1.91 (d, J = 1.4 Hz, 3H),

1.43 (s, 3H), 1.36 (s, 3H), 1.06 (s, 9H), 0.92 - 0.87 (m, 6H)

C7oHi₁₈N₆0₁₆S₂ caled [M + H]⁺ = 1361.80 amu; found m / z = 1362.26.

Example 142

HS

OH (142)

The compound of Example 141 is reduced according to the methods below to produce the compound in question.

302

Example 143

<img file="MX368258B_D0205.tif" />

(143)

(S, E) -4- ((S) -2- ((S) -3- (3 - ((14-mercapto-3,6,9,12 tetraoxatetradecyl) oxy) phenyl) -3-methyl-2 acid (methylamino) butanamido) -N, 3,3-trimethylbutanamide) -2,5-dimethylhex-2-oenic

The title compound was prepared by saponification, subsequently the TEA promoted the elimination of Boc, according to the exact methods described in Nieman et al. from (6S, $ 9, 12S, E) -ethyl 9- (tert-butyl) -12-isopropyl-

2,2,5,11,14-pentamethyl-4,7,10-trioxo-6- (2- (3 - ((16-oxo-

3,6,9,12-tetraoxa-15-thiaheptadecyl) oxy) phenyl) propan-2-yl) -3oxa-5,8, ll-triazapentadec-13-en-15-oato (56 mg, 0.029 mmol) for give the title compound (43 mg, 82%) as a whitish foam after complete removal of excess ASD.

And NMR (400 MHz, Methanol-d₄) δ (ppm) 8.48 (d, J = 8.3 Hz, 1H),

7.47 - 7.29 (m, 1H), 7.21 - 7.04 (m, 1H), 6.95 (t, J = 9.4 Hz 1H), 6.80 (d, J = 9.7 Hz, 1H), 5.08 (t, J = 10.1 Hz, 1 HOUR) ,

4.97 - 4.94 (m, 1H), 4.

- 3.82 (m, 2H), 3.80 J = 6.4 Hz, 2H), 2.53 (

J = 1.4 Hz, 3H), 1.47

0.93 (dt, J = 11.2, 3.4

1449.85 amu; found

303 (s, 1H), 4.24 - 4.13

3.58 (m, 14H), 3.17 (s, s, 3H), 2.11-2.03 (m, (s, 3H), 1.40 (s, 3H),

Hz, 15H). C74H124N6O18S2 cm / z = 1450.06.

^: «: 1 unit w 1 (m, 2H), 3.95

3H), 2.92 (t,

1H), 1.94 (d,

1.09 (s, 9H), alcd. [M + H]⁺ =

Example 144

<img file="MX368258B_D0206.tif" />

(144)

The compound of Example 143 is reduced according to the methods below to produce the compound in question

Example 145

<img file="MX368258B_D0207.tif" />

(145) (mAb-SPDP-Compound 142) produced using the synthesis method of Compound 142, above, and the SPDP binding method described below.