Intermediates for the preparation of bivalent smac mimetics

[0001] The present application claims priority from U.S. Provisional Patent Application No. 60/798 018, filed May 5, 2006, and U.S. Provisional Patent Application No. 60/923 415, filed April 13, 2007.

Scope of the invention [0002] The invention relates to the field of therapeutic chemistry. In particular, the invention relates to intermediates for the preparation of bivalent Smac mimetics. The latter act as inhibitors of the protein inhibitor of apoptosis. Also disclosed is the use of these mimetics to induce or sensitize cells to induce apoptotic cell death.

Background Art [0003] The aggressive cancer cell phenotype is the result of many genetic and epigenetic changes leading to deregulation of intracellular signaling pathways (Ponder, Nature 411: 336 (2001)). Common to all cancer cells, however, is their inability to perform an apoptosis program, and the lack of adequate apoptosis due to defects in a normal apoptosis machine is a special sign of cancer (Lowe et al., Carcinogenesis 21: 485 (2000)). Most current cancer therapies, including chemotherapeutic agents, radiation and immunotherapy, act indirectly to induce apoptosis in cancer cells. The inability of cancer cells to perform an apoptotic program due to defects in normal apoptotic machinery is therefore often associated with an increase in resistance to chemotherapy, radiation or immunotherapy induced apoptosis. Primary or acquired resistance of human cancer of various origins to current treatment protocols due to defects in apoptosis is a major problem in current cancer therapy (Lowe et al., Carcinogenesis 21: 485 (2000); Nicholson, Nature 407: 810 (2000)). Accordingly, current and future efforts to design and develop new molecular-specific anti-cancer therapies to improve the survival and quality of life of cancer patients must include strategies that are specifically targeted at cancer cell resistance to apoptosis. In this regard, targeting critical negative regulators that play a major role in directly inhibiting apoptosis in cancer cells represents a highly promising therapeutic strategy for the design of a new anti-cancer drug.

[0004] Two classes of central negative regulators of apoptosis have been identified. The first class of regulators is the Bcl-2 family of proteins, for example two strong anti-apoptotic molecules, Bcl-2 and Bcl-XL proteins (Adams et al., Science 281: 1322 (1998); Reed, Adv. Pharmacol. 41: 501 (1997) ; Reed et al., J. Cell. Biochem. 60:23 (1996)). A broad review of therapeutic strategies targeting Bcl-2 and Bcl-XL in cancer to restore cancer cell sensitivity and overcome cancer cell resistance to apoptosis (Adams et al., Science 281: 1322 (1998); Reed, Adv. Pharmacol. 41: 501 (1997); Reed et al., J. Cell. Biochem. 60:23 (1996)). Several laboratories have become interested in the design of small molecule inhibitors of Bcl2 and Bcl-XL.

[0005] The second class of central negative regulators of apoptosis is an apoptosis protein inhibitor (IAP) (Deveraux et al., Genes Dev. 13: 239 (1999); Salvesen et al., Nat. Rev. Mol. Cell. Biol. 3: 401 (2002)). This class includes proteins such as XIAP, cIAP-1, cIAP-2, ML-IAP, HIAP, KIAP, TSIAP, NAIP, survivin, livin, ILP-2, apollon and BRUCE. IAP proteins strongly suppress apoptosis induced by a wide variety of apoptotic stimuli, including chemotherapeutic agents, radiation and immunotherapy in cancer cells.

[0006] X-related IAP (XIAP) is the most potent inhibitor of suppressing apoptosis among all IAP members (Holcik et al., Apoptosis 6: 253 (2001); LaCasse et al., Oncogene 17: 3247 (1998) ; Takahashi et al., J. Biol. Chem. 273: 7787 (1998); Deveraux et al., Nature 388: 300 (1997); Sun et al., Nature 401: 818 (1999); Deveraux et al., EMBO J. 18: 5242 (1999); Asselin et al., Cancer Res. 61: 1862 (2001). XIAP plays a key role in the negative regulation of apoptosis in death receptor and mitochondrial mediated pathways. XIAP acts as a powerful endogenous inhibitor of apoptosis by directly binding and strongly inhibiting three enzymes, members of the caspase family, caspase-3, -7, and -9 (Takahashi et al., J. Biol. Chem. 273: 7787 (1998); Deveraux et al., Nature 388: 300 (1997); Sun et al., Nature 401: 818 (1999); Deveraux et al., EMBO J. 18: 5242 (1999); Asselin et al., Cancer Res. 61: 1862 (2001); Riedl et al., Cell 104: 791 (2001); Chai et al., Cell 104: 769 (2001); Huang et al., Cell 104: 781 (2001)). XIAP contains three baculovirus domains of the apoptosis inhibitor (BIR) repeating motif as well as the RING C-terminal finger. The third BIR domain (BIR3) is selectively targeted to caspase-9, initiating caspase in the mitochondrial pathway, while the linker region between BIR1 and BIR2 inhibits both caspase-3 and caspase-7 (Salvesen et al., Nat. Rev. Mol Cell Biol. 3: 401 (2002)). Although binding to XIAP prevents the activation of all three caspases, it is obvious that interaction with caspase9 is most critical for its inhibition of apoptosis (Ekert et al., J. Cell Biol. 152: 483 (2001); Srinivasula et al., Nature 410: 112 (2001)). Because XIAP blocks apoptosis in a later executive phase, at which many signaling pathways converge, XIAP-targeted strategies may be particularly effective in overcoming cancer cell resistance to apoptosis (Fulda et al., Nature Med. 8: 808 (2002); Arnt et al., J. Biol. Chem. 277: 44236 (2002)).

[0007] Although the exact role of XIAP in any type of cancer is far from being fully understood, there is growing evidence indicating that XIAP is widely overexpressed in many types of cancer and may play an important role in resistance of cancer cells to many current therapeutic agents (Holcik et al. , Apoptosis 6: 253 (2001); LaCasse et al., Oncogene 17: 3247 (1998)).

[0008] The XIAP protein has been found to be expressed in most NCI 60 human cancer cell lines (Tamm et al., Clin. Cancer Res. 6: 1796 (2000)). Analysis of tumor samples in 78 previously untreated patients showed that those with lower XIAP levels showed significantly longer survival (Tamm et al., Clin. Cancer Res. 6: 1796 (2000)). XIAP has been found to be expressed in human malignant glioma (Wagenknecht et al., Cell Death Differ. 6: 370 (1999); Fulda et al., Nature Med. 8: 808 (2002)). XIAP has been found to be expressed in human prostate cancer cells and blocks ligand-related Apo2 / tumor necrosis factor-induced apoptosis inducing ligand-mediated apoptosis in prostate cancer cells in the presence of mitochondrial activation (McEleny et al., Prostate 51: 133 (2002); Ng et al., Mol. Cancer Ther. 1: 1051 (2002)). XIAP is overexpressed in non-small cell lung cancer (NSCLC) in patients and has been associated with NSCLC pathogenesis (Hofmann et al., J. Cancer Res. Clin. Oncol. 128: 554 (2002)). XIAP expression and no downregulation of XIAP on cisplatin treatment have been associated with cisplatin resistance of human ovarian cancer (Li et al., Endocrinology 142: 370 (2001); Cheng et al. Drug Resist. Update 5: 131 (2002)). Taken together, these data suggest that XIAP may play an important role in the resistance of several human cancers to current therapeutic agents.

[0009] Apoptosis is not a single process, rather it is involved in many different, sometimes interrelated signaling pathways leading to cell degradation. The pathways associated with a particular form of apoptosis depend on many factors, such as injury or injuries that initiate the process. Other factors include activation or over-activation of specific receptors, such as activation of "death" receptors by tumor necrosis factor alpha (TNFa), associated with tumor necrosis factor

Apoptosis-inducing ligand (TRAIL or Apo2L), or FAS ligand. The other determining factor is the type of cell involved, because different signaling pathways for so-called type I and type II cells are presented after activation of the Fas or TNFα receptor.

[0010] TRAIL (Apo2L) has been shown to be a selective and potent inducer of apoptosis in cancer (but not normal) cells after binding to any of the two pro-apoptotic TRAIL receptors, TRAILR1 (or DR4) (Pan et al., Science 276: 111 (1997)) or TRAIL-R2 (KILLER, or DR5) (Wu et al., Nat. Genet. 17: 141-143 (1997); Pan et al., Science 277: 815 (1997); Walczak et al. ., EMBO J. 16: 5386 (1997)). Activation of pro-apoptotic death receptors by TRAIL induces the formation of the death inducing signaling complex (DISC), which consists of the FADD receptor as an adapter (Kischkel et al., Immunity 12: 611 (2000); Kuang et al., J. Biol. Chem 275: 25065 (2000)), and caspase-8 as the initiating caspase. After DISC formation, caspase-8 is self-processed and activated by induced neighborhood (Medema et al., EMBO J. 16: 2794 (1997); Muzio et al., J. Biol. Chem. 273: 2926 (1998)).

[0011] TRAIL has attracted significant attention as a potential cancer drug (French et al., Nat. Med. 5: 146 (1999)) due to its selective targeting of cancer cells, while most normal cells appear to be TRAIL resistant. (Ashkenazi et al., Science 281: 1305 (1998); Walczak et al., Nat. Med. 5: 157 (1999)). Systemic administration of TRAIL has been shown to be safe and effective in killing breast or colorectal xenograft tumors and prolonged survival in mice (Walczak et al. Nat. Med. 5: 157 (1999)). Although TRAIL can specifically kill many types of cancer cells, many others are resistant to TRAIL (Kim et al., Clin. Cancer Res. 6: 335 (2000); Zhang et al., Cancer Res. 59: 2747 (1999)). In addition, cancer cells are killed by the use of antibodies (monoclonal or polyclonal) that specifically recognize TRAIL-R1 or TRAIL-R2.

[0012] Numerous mechanisms have been identified as potential factors responsible for TRAIL resistance. Such mechanisms exist at many levels, including receptors, mitochondria, post-mitochondrial and DISC. For example, loss of caspase-8 expression (Teitz et al., Nat. Med 6: 529 (2000); Griffith et al., J. Immunol. 161: 2833 (1998)), or high expression of the cellular protein FLICE inhibitor (cFLIP) (Kim et al., Clin. Cancer Res. 6: 335 (2000); Zhang et al., Cancer Res. 59: 2747 1999; Kataoka et al., J. Immunol. 161: 3936 (1998)) make cancer cells resistant to TRAIL. Yeh et al. have shown that embryonic mouse fibroblasts deficient in cFLIP are particularly susceptible to receptor-mediated apoptosis (Yeh et al., Immunity 12: 533 (2000)). Several splicing cFLIP variants are known, including the short splicing variant, cFLIP-S, and the longer splicing variant, cFLIP-L. Embryonic mouse cFLIP-deficient fibroblasts have been shown to become resistant to TRAIL-induced apoptosis as a result of retrovirally mediated cFLIP-S transduction (Bin et al., FEBS Lett. 510: 37 (2002)).

[0013] Although TRAIL is a potentially promising candidate for tumor selective death receptor activation (i.e., induces apoptosis preferentially in tumor cells, but not in normal tissues), many cancer cells are resistant to apoptosis-inducing drugs as discussed above. As a result, treatment with such drugs often requires simultaneous radiation therapy and / or cytotoxic chemical agents to achieve a therapeutic effect. However, both radiation and chemotherapy have significant side effects, and are generally avoided if possible.

[0014] Thus, there is a need for an agent that can selectively and efficiently sensitize tumor cells to specific apoptosis-inducing drugs such as TRAIL or TRAIL receptor antibodies, without sensitizing surrounding normal cells at the same time. Such an agent would also be useful for limiting or preventing drug resistance typically associated with receptor-mediated use

EP 2 019 671 B1 to apoptotic cancer drugs, improving their effectiveness and eliminating the need for combination therapies.

[0015] Recently, Smac / DIABLO (the second mitochondrial caspase activator) has been identified as a protein released from the mitochondria to the cytosol in response to an apoptotic stimulus (Budihardjo et al., Annu. Rev. Cell Dev. Biol. 15: 269 (1999) ; Du et al., Cell 102: 33 (2000)). Smac is synthesized with an N-terminal mitochondrial targeting sequence that is proteolytically removed during maturation into the mature polypeptide. Smac has been shown to interact directly with XIAP and other IAPs and breaks their binding to caspases and facilitates caspase activation. Smac is a potent endogenous XIAP inhibitor.

[0016] Recently, high-resolution, experimental three-dimensional (3D) structures of the XIAP BIR3 domain in complex with Smac protein and peptide have been determined (Sun et al., J. Biol. Chem. 275: 36152 (2000); Wu et al., Nature 408: 1008 (2000)) (Figure 1). The Smac N-terminal tetrapeptide (Ala-Val-Pro-Ile, or AVPI (SEQ ID NO: 1)) recognizes the surface furrow on the XIAP BIR3 domain by several hydrogen bonding and van der Waals interactions. The interaction between BIR3 and caspase-9 has also been shown to involve four residues (Ala-Thr-Pro-Phe, or ATPF (SEQ ID NO: 2)) at the amino terminus of the small caspase-9 subunit to the same surface furrow in the domain BIR3. Several recent studies have convincingly demonstrated that Smac promotes the catalytic activity of caspase-9 by competing with caspase-9 for the same furrow binding on the surface of the BIR3 domain (Ekert et al. J. Cell Biol. 152: 483 (2001); Srinivasula et al., Nature 410: 112 (2001)).

[0017] Unlike protein-protein interactions, Smac-XIAP interaction is mediated by only four amino acid residues on the Smac protein and a well-defined surface furrow on the BIR3 domain

XIAP. K value_d AVPI Smac peptide (SEQ ID NO: 1) versus XIAP (K_d = 0.4 μΜ) is essentially the same as mature Smac protein (K_d = 0.42 μΜ). This well-defined interaction site is ideal for designing non-peptide, drug-like small molecules that mimic the binding of Smac to XIAP.

[0018] It has recently been shown that the Smac peptide that is cell-permeated, which consists of the first four amino acid residues (AVPI (SEQ ID NO: 1)) from the N Smac end bound to the carrier peptide to facilitate intracellular delivery, sensitizes various cancer cells in in vitro and malignant glioma cells in vivo for apoptosis induced by death receptor ligation or cytotoxic drugs (Fulda et al. Nature Med. 8: 808 (2002)). Importantly, this Smac peptide strongly enhanced the anti-tumor activity of Apo2L / TRAIL in a malignant glioblastoma xenograft in vivo model. Complete destruction of established tumors and mouse survival was only achieved with the combination of Smac and Apo2L / TRAIL peptide therapy. Importantly, the Smac peptide does not show detectable toxicity to normal brain tissue.

[0019] A second recent independent study also showed that peptides composed of the first four to eight N-terminal Smac amino acid residues associated with various carrier peptides increase the induction of apoptosis and long-term antiproliferative effects of various chemotherapeutic drugs, including paclitaxel, etoposide, SN-38, and doxorubicin in MCF-7 and other human breast cancer cell lines (Amt et al., J. Biol. Chem. 277: 44236 (2002). This study firmly showed that XIAP and cIAP-1 are the primary molecular targets for these peptides in cells.

[0020] A third study showed that the Smac peptide of the first seven N-terminal polyarginine-bound residues restored apoptosome activity and reversed resistance to apoptosis in H460 non-small cell lung cancer cells (Yang et al., Cancer Res. 63: 831 (2003)) . XIAP has been shown to respond4

EP 2 019 671 B1 is responsible for the defect in apoptosome activity and suppression of caspase activity in H460 cells. When used in combination with chemotherapy, the cell-permeable Smac peptide reversed tumor growth in vivo with little toxicity in mice. Taken together, this latest independent study strongly suggests that strong, persistent, cell-mediated Smac mimetic may have great therapeutic potential in the treatment of human breast cancer and other types of cancer.

[0021] Peptide-based inhibitors are useful tools for explaining the antiapoptotic activity of IAP and the role of IAP in cancer cell response to chemotherapeutic agents. But peptide-based inhibitors in general have internal limitations as potentially useful therapeutic agents. These restrictions include poor cell permeability and poor in vivo stability. Indeed, in these three published studies using Smac-based peptide inhibitors, peptides had to be condensed with carrier peptides to be relatively permeable to the cell.

[0022] Published US Application No. 2005/0197403 discloses Smac dimeric mimetic compounds of formula I:

<td>R¹' AND</td><td></td><td>R⁷</td>

<td>ID? O ^^ R³ |</td><td> -_L-</td><td>%AND_ABOUT</td>

<td></td><td></td><td>| v y / ^ NH</td>

<td>R⁷'</td><td></td><td>AND Βί</td>

1 'in which R¹ and R^1' are selected from hydrogen, optionally substituted methyl and hydroxyl;

R² and R²'are selected from optionally substituted methyl and optionally substituted ethyl;

R³ and R³'are selected from CH2, NH, O and S;

R4 and R4 'are selected from CH and N;

R⁵-R⁸, and R⁵'-R⁸'are selected from hydrogen, optionally hetero-, optionally substituted alkyl, optionally hetero-, optionally substituted alkenyl, optionally hetero-, optionally substituted alkynyl, optionally hetero-, optionally substituted aryl; and

L is a linker covalently linking R², R⁵, R⁶ or R⁷, with R²', R⁵', R⁶'or R⁷'or a pharmaceutically acceptable salt thereof.

[0023] US 2005/0197403 discloses mono- and dimeric compounds that clearly do not relate to the invention as defined in the claims.

[0024] The problem underlying the claims is the need for compounds that are useful intermediates for the production of IAP inhibitors as given in paragraph [0058] on page 21 of the description.

5c

The provided solution is a compound having formula XIII as defined in claim 1 wherein R^5cmeans NCOR⁸ or NCO2R⁸. A specialist in this field would have no reason to modify the relationship with

WO-A-2005/069894, WO-A-2006/010118, or Sun et al., "STRUCTURE-BASED DESIGN OF POTENT, CONFORMATIONALLY CONSTRAINED SMAC MIMETICS", JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, vol. 126, No. 51, pp. . 16686-16687 &amp; S1-S20 containing the CH2 group at R^5c to reach a relationship with the claims, without the benefit of subsequent information from this application.

EP 2 019 671 B1

SUMMARY OF THE INVENTION [0025] The invention is set out in the appended claims. Embodiments of the description that do not fall within the scope of these claims are for illustrative purposes only and do not form part of the present invention.

[0026] It is generally accepted that the inability of cancer cells or their supporting cells to undergo apoptosis in response to genetic damage or exposure to factors causing apoptosis (such as anti-cancer agents and radiation) is a major factor in the progression and progression of cancer. Induction of apoptosis in cancer cells or their supporting cells (e.g., new vascular cells in tumor vasculature) is considered to be the universal mechanism of action of almost all effective cancer drugs or radiation therapy on the market or in today's practice. One reason for the cell's inability to undergo apoptosis is increased expression and accumulation of IAPs.

[0027] The present description includes that exposure of animals suffering from cancer or other hyperproliferative disorders or diseases associated with dysregulation of apoptosis to therapeutically effective amounts of the drug (drugs) (e.g., small molecules) that inhibit IAP function (s) will result in complete killing diseased cells or support cells (those cells whose continuity of survival depends on hyperactivity or overexpression of IAPs) and / or will cause that such cells as a population will be more susceptible to cell death induction activity for cancer drugs or radiation treatments. The present description considers that IAP inhibitors meet the unmet need for treatment of many types of cancer when they are administered as monotherapy to induce apoptosis in IAP function dependent cancer cells, or when they are administered in a time-dependent manner with other cell-inducing cancer drugs or radiation treatments. . to confer a greater proportion of cancer cells or support cells susceptible to an apoptosis program compared to the corresponding portion of cells in an animal treated with only a cancer drug or only radiation therapy.

[0028] Combination therapy of animals with a therapeutically effective amount of a compound of the present description and treatment with an anti-cancer agent or radiation may result in a stronger tumor response and clinical benefit in such animals compared to the treated compound alone or the anti-cancer drugs / radiation alone. In other words, because the compounds lower the apoptotic threshold of all cells that express IAP, the proportion of cells that successfully perform an apoptosis program in response to apoptosis-inducing anti-cancer drug / radiation activity is increased. Alternatively, the compounds of the present description may be used to allow administration of lower, and therefore less toxic and more tolerated, doses of the anti-cancer agent and / or radiation to obtain the same tumor response / clinical benefit than the conventional anti-cancer agent / radiation dose alone. Since the doses of all approved anti-cancer drugs and radiation therapy are known, the present description includes various combinations thereof with the compounds of the present invention. Also, because the compounds of the present description act at least in part by inhibiting IAP, exposing cancer cells and support cells to therapeutically effective amounts of compounds may be temporarily associated to coincide with cell attempts to program an apoptosis in response to an anti-cancer agent or radiation therapy. Thus, administration of the compositions of the present description in connection with certain time dependencies, provides especially effective therapeutic practices.

[0029] The present description relates to Smac mimetics that are useful for inhibiting the activity of IAP proteins and increasing the sensitivity of cells to agents that cause apoptosis. For example, Smac mimetics are compounds of formula II:

EP 2 019 671 B1

<td colspan="2">^D'\</td><td></td><td>AND</td>

<td colspan="2">Ά - ( in \</td><td></td><td>J</td>

<td colspan="2">and₂ about</td><td colspan="2">Here</td>

<td></td><td colspan="3"></td>

<td rowspan="2">^x \ ------</td><td colspan="3">D &lt; / 1 ·</td>

<td></td><td>at</td><td></td>

<td colspan="3">1 rt '' "'" ^NT Ki A₂- {j</td><td>\ HERE'</td>

in which:

A1 and A1 'are independently selected from the group consisting of hydrogen, optionally substituted alkyl and Z;

A2 and A2 'are independently selected from the group consisting of hydrogen, optionally substituted alkyl and COR¹where A2 is not present when V is O and A2 'is not present when V' is O;

V and V 'are independently selected from the group consisting of N, CH and O;

W and W 'are independently selected from the group consisting of CH and N;

X and X 'are independently optionally substituted C 1-3 alkyl;

Y and Y 'are independently selected from the group consisting of CONR¹, C (O) O, (CR¹R²) 1-3, where one or more CH2 groups can be replaced by O, S or NR¹, optionally substituted aryl and optionally substituted heteroaryl;

D and D 'are independently selected from the group consisting of optionally substituted alkenyl and (CR¹R²) n^5a(CR³R⁴) M;

J and J 'are independently selected from the group consisting of optionally substituted alkenyl and (CR¹R²) p^5b15 (CR³R⁴) Q;

T and T 'are independently selected from the group consisting of C = O, C = S, C = NR¹, S, O, NR¹, CR¹R², optionally substituted carbocycle, optionally substituted heterocycle, optionally substituted aryl and optionally substituted heteroaryl;

U and U 'are independently selected from the group consisting of hydrogen, NR¹R², OR¹, SR¹, optionally substituted alkyl and optionally substituted aryl;

n, m, and qq are independently 0-5;

each R¹ is selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted carbocycle, optionally substituted heterocycle, optionally substituted aryl, optionally substituted heteroaryl and Z;

3 4 each R², R³ and R⁴ is independently selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted carbocycle, optionally substituted heterocycle, optionally substituted aryl and optionally substituted heteroaryl;

EP 2 019 671 B1

R^5a and R^5b are independently selected from the group consisting of C = O, C = S, C = NR¹, S, O, NR¹ and CR¹R²; and

Z is a linker covalently linking one of A1, Y, D, J, T and U with one of A1 ', Y', D ', J', T 'and U';

or pharmaceutically acceptable salts or prodrugs thereof.

[0030] The description relates to compounds represented by formula II which are inhibitors of IAP proteins. The description relates to the use of compounds of the invention for inducing apoptosis in cells. The description also relates to the use of compounds for sensitizing cells to apoptosis inducing agents. The compounds are useful for treating, alleviating or preventing disorders responsive to the induction of apoptotic cell death, e.g., disorders characterized by dysregulation of apoptosis, including hyperplasia diseases such as cancer. The compounds can be used to treat, alleviate or prevent cancer that is characterized by resistance to cancer therapies (e.g., those that are resistant to chemicals, resistant to radiation, resistant to hormones, and the like). The compounds can be used to treat hyperplasia diseases characterized by IAP overexpression.

[0031] The present description provides pharmaceutical compositions comprising a compound of formula II in a therapeutically effective amount for inducing apoptosis in cells or for sensitizing cells to agents that cause apoptosis.

[0032] The description further provides kits comprising a compound of formula II and instructions for administering the compound to an animal. The kits may optionally contain other therapeutic agents, e.g., anti-cancer agents or agents that modulate apoptosis.

BRIEF DESCRIPTION OF THE DRAWINGS [0033]

Figure 1 shows the competitive binding of Smac mimetics to the BIA3 XIAP protein.

Figure 2 shows the inhibition of cell growth in MDA-MB-231, MAMLE-3M, SK-OV-3 and OVCAR-4 cells by SH-164.

Figure 3 shows the induction of cell death in MDA-MB-231, MAMLE-3M, and OVCAR-4 cells by SH-164.

Figures 4A-4C show inhibition of cell growth in MDA-MB-231 (A), MDA-MB-453 (B), and PC-3 (C) cells by SH-164 in combination with TRAIL.

Figure 5 shows inhibition of cell growth in MDA-MB-231 cells by SH-164 in combination with cisplatin or mitoxantrone.

Figure 6 shows the induction of apoptosis by SH-164 in MDA-MB-231 breast cancer cell lines.

DETAILED DESCRIPTION OF THE INVENTION [0034] The invention is set out in the appended claims. Embodiments of the description that do not fall within the scope of these claims are for illustrative purposes only and do not form part of the present invention. The present description relates to divalent conformationally limited compounds represented by formula II which are Smac mimetics and act as IAP inhibitors. These compounds sensitize cells to apoptosis-inducing factors and in some cases induce apoptosis themselves by inhibiting IAP. Thus, the description relates to methods for sensitizing cells to apoptosis-inducing agents and methods for inducing apoptosis in cells, comprising contacting the cells with a compound of formula II alone or in combination with an apoptosis-causing agent. The following description relates to methods of treating, alleviating or preventing disorders in an animal that respond to induction of apoptosis, comprising administering to the animal a compound of formula II and an apoptosis-inducing agent. Such disorders include disorders characterized by dysregulation of apoptosis and disorders characterized by IAP overexpression.

[0035] The term "IAP protein" as used herein refers to any known member of the apoptosis inhibitor protein family, including, but not limited to, XIAP, cIAP-1, cIAP-2, MLIAP, HIAP , TSIAP, KIAP, NAIP, survivin, livin, ILP-2, apollon and BRUCE.

[0036] The term "IAP overexpression", as used herein, refers to elevated levels (e.g., abnormal levels) of mRNAs encoding IAP protein (s), and / or elevated levels of IAP protein (s) in cells compared to similar suitable non-pathological cells expressing basal levels of mRNA encoding IAP proteins or having basal levels of IAP proteins. Methods for detecting mRNA levels encoding IAP proteins or IAP protein levels in a cell include, but are not limited to, Western hybridization using IAP protein antibodies, immunohistochemical methods, and methods of nucleic acid amplification or direct RNA detection. As important as the absolute level of IAP proteins in cells to determine if they overexpress IAP proteins is also the relative level of IAP proteins relative to other pro-apoptotic signaling molecules (e.g., the pro-apoptotic Bcl-2 family of proteins) in such cells. When the balance of these two is such that, were it not for IAP protein levels, pro-apoptotic signaling molecules would be sufficient to cause the cells to undergo apoptosis and death, survival of these cells would be dependent on IAP proteins. In such cells, exposure to an inhibitory effect of an effective amount of an IAP protein inhibitor will be sufficient to cause the cell to undergo apoptosis and death. Thus, the term "IAP protein overexpression" also refers to cells that, due to relative levels of pro-apoptotic and anti-apoptotic signals, undergo apoptosis in response to inhibitory effective amounts of compounds that inhibit the action of IAP proteins.

[0037] The terms "anti-cancer agent" and "anti-cancer drug" as used herein refer to any therapeutic agents (e.g., chemotherapeutic compounds and / or molecular therapeutic compounds), radiation therapy, or surgery used to treat hyperplasia diseases such as cancer (e.g., in mammals).

[0038] The term "prodrug," as used herein, refers to a pharmacologically inactive derivative of a parent drug "drug" that requires biological (e.g., spontaneous or enzymatic) transformation in a target physiological system for release or transformation (e.g., enzymatically , physiologically, mechanically, electromagnetically) of a pro-drug into the active drug. Prodrugs are designed to overcome problems associated with persistence, toxicity, lack of specificity, or limited bioavailability. Exemplary prodrugs include the active drug molecule itself and a chemical masking group (e.g., a group that reversibly suppresses drug activity). Certain preferred prodrugs are variants or derivatives of compounds that have groups useful for cleavage under metabolic conditions. Exemplary prodrugs become pharmaceutically active in vivo or in vitro when they undergo solvolysis under physiological conditions or undergo enzymatic degradation or other biochemical transformation (e.g., phosphorylation, hydrogenation, dehydrogenation, glycosylation). Prodrugs often have the advantages of solubility, tissue compatibility, or delayed release in the body of a mammal. (see, e.g., Bundgard, Design of Prodrugs, p. 7-9, 21-24, Elsevier, Amsterdam (1985); and Silverman, Organic Chemistry of Drug Design and Drug Action, pp. 352-401, Academic Press, San Diego, CA (1992). Common prodrugs include acid derivatives such as esters prepared by the reaction of parent acids with the appropriate alcohol (e.g., lower alkanol), amides produced by the reaction of the parent acid compound with an amine, or basic groups reacted to form an acylated basic derivative (e.g., lower alkylamide) ).

[0039] The term "pharmaceutically acceptable salt," as used herein, refers to any salt (e.g., obtained by reaction with an acid or base) of a compound of the present invention that is physiologically tolerated in the target animal ( e.g., mammalian). The salts of the compounds herein may be derived from inorganic or organic acids and bases. Examples of acids include, but are not limited to, hydrochloric, hydrobromic, sulfuric, nitric, perchloric, fumaric, maleic, phosphoric, glycolic, lactic, salicylic, succinic, toluene-p-sulfonic, tartaric, acetic, citric, methanesulfonic, ethanesulfonic , formic, benzoic, malonic, sulfonic, naphthalene-2-sulfonic, benzenesulfonic, and the like. Other acids, such as oxalic, although not themselves pharmaceutically acceptable, may be used in the preparation of salts useful as intermediates in the preparation of compounds of the description and their pharmaceutically acceptable acid addition salts.

[0040] Examples of the base include, but are not limited to, alkali metal (e.g., sodium) hydroxides, alkaline earth metal (e.g., magnesium) hydroxides, ammonia and compounds of formula NW4⁺wherein W is C1-4 alkyl, and the like.

[0041] Examples of salts include, but are not limited to: acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphors, camphorsulfonate, cyclopentanopropionate, digluconate, dodecyl sulfate, ethanesulfonate, fumarate, flucoheptanoate, glycerophosphate, heptophosphate, heptulfate hydroxyethanesulfonate, lactate, maleate, mesylate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, oxalate, pamoate, pectinate, persulfate, phenylpropionate, picrate, pivalate, propionate, succinate, tartrate, thiocyanate, tosylate, undecanoate, and the like. Other examples of salts include the anions of the compounds of the present invention associated with a suitable cation such as Na⁺, NH4⁺ and NW4⁺ (where W is a C1-4 alkyl group) and the like. For therapeutic use, the salts of the compounds of this specification are considered pharmaceutically acceptable. However, non-pharmaceutically acceptable acid and base salts may also find use, for example, in the preparation or purification of a pharmaceutically acceptable compound.

[0042] The term "therapeutically effective amount", as used herein, refers to an amount of a therapeutic agent sufficient to alleviate one or more symptoms of the disorder, or to prevent the progression of the disorder, or to cause regression of the disorder. For example, with respect to the treatment of cancer, a therapeutically effective amount preferably refers to an amount of a therapeutic agent that reduces the rate of tumor growth, reduces the tumor mass, reduces the number of metastases, increases the time to tumor progression, or increases the survival time by at least 5%, preferably at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 100% .

[0043] The terms "sensitize" and "sensitizing" as used herein refer to the administration, by administering a first agent (e.g., a compound of formula II), to an animal or cell in an animal being more susceptible or more responsive to effects biological (e.g., promoting or delaying an aspect of cellular function, including, but not limited to, cell division, growth, proliferation, infiltration, angiogenesis or apoptosis of a cell) of the second agent. The sensitizing effect of the first agent on the target cell may be measured as a difference in the intended biological effect (e.g., promoting or delaying aspect of cellular function, including, but not limited to, growth, proliferation, infiltration, angiogenesis or cell apoptosis) observed when administered second agent with administration

EP 2 019 671 B1 first agent and without it. The sensitized cell response may be increased by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90 %, at least 100%, at least 150%, at least 200%, at least 350%, at least 300%, at least 350%, at least 400%, at least 450%, or at least 500% of the responses absence of first measure.

[0044] The term "dysregulation of apoptosis" as used herein refers to any abnormality in the ability (e.g., predisposition) of a cell to undergo cell death by apoptosis. Dysregulation of apoptosis is associated or induced by many conditions, including, for example, autoimmune disorders (e.g., systemic lupus erythematosus, rheumatoid arthritis, graft versus host reaction, myasthenia gravis or Sjogren's syndrome), chronic inflammation (e.g., psoriasis, asthma or Crohn's disease), excessive growth disorders (e.g., tumors, B-lymphomas, or T-lymphomas), viral infections (e.g., herpes, papilloma or HIV), and other conditions such as osteoarthritis and atherosclerosis. It should be noted that when deregulation is induced or associated with a viral infection, the viral infection may or may not be detectable at the time of occurrence or observation of deregulation. This means that virus-induced dysregulation can occur even after the symptoms of the viral infection have disappeared.

[0045] The term "hyperplasia disease," as used herein, refers to any condition in which the localized population of proliferating cells in an animal is not regulated by normal restrictions of normal growth. Examples of hyperproliferative disorders include, but are not limited to, tumors, tumors, lymphomas and the like. Cancer is called benign if it is not infiltrated or metastasized, and malignant if it causes any of them. A "metastatic" cell means that the cell can infiltrate and destroy neighboring body structures. Excessive hyperplasia is a form of cell proliferation involving an increase in the number of cells in a tissue or organ without significantly altering structure or function. Metaplasia is a form of controlled cell growth in which one type of fully differentiated cell changes to another type of differentiated cell.

[0046] Pathological growth of activated lymph cells often leads to an autoimmune disorder or chronic inflammation. As used herein, the term "autoimmune disorder" refers to any condition in which the body produces antibodies or immune cells that recognize the body's own molecules, cells or tissues. Non-limiting examples of autoimmune disorders include autoimmune hemolytic anemia, autoimmune hepatitis, Berger's disease or IgA nephropathy, celiac disease, chronic fatigue syndrome, Crohn's disease, dermatomyositis, fibromyalgia, graft versus host disease, Basedow's disease, Histimist thyroiditis itself thrombocytopenic purpura, lichen planus, multiple sclerosis, myasthenia gravis, psoriasis, rheumatic fever, rheumatic arthritis, scleroderma, Sjogren's syndrome, systemic lupus erythematosus, type 1 diabetes, ulcerative colitis, acquired vitiligo, and the like.

[0047] The term "cancer," as used herein, refers to any abnormal growth of benign (non-cancerous) or malignant (cancerous) cells.

[0048] The term "anti-cancer agent," as used herein, refers to any compound that delays the growth, growth, or spread of a target (e.g., malignant) tumor. [0049] The terms "prevent", "preventing" and "prevention" as used herein refer to a decrease in the occurrence of pathological cells (e.g., overgrowing or cancerous

EP 2 019 671 B1) in an animal. Prevention can be complete, e.g., complete absence of pathological cells in an individual. Prevention may also be partial, such that the occurrence of pathological cells in a subject is less common than occurrence without the present invention.

[0050] The term "agents that modulate apoptosis," as used herein, refers to agents that are involved in modulating (e.g., inhibiting, reducing, increasing, promoting) apoptosis. Examples of agents that modulate apoptosis include proteins that contain the death domain, such as, but not limited to, Fas / CD95, TRAMP, TNF RI, DR1, DR2, DR3, DR4, DR5, DR6, FADD, and RIP.

Other examples of agents that modulate apoptosis include, but are not limited to, TNFα, Fas ligand, antibodies to Fas / CD95 and other TNF family receptors, TRAIL (also known as Ligand Apo2 or Apo2L / TRAIL), agonists (e.g., monoclonal or polyclonal agonist antibodies) TRAIL-R1 or

TRAIL-R2, Bcl-2, p53, BAX, BAD, Akt, CAD, PI3 kinases, PP1 and protein caspases. Modulating agents broadly include agonists and antagonists of TNF family receptors and TNF family ligands. Agents that modulate apoptosis may be soluble or membrane bound (e.g., ligand or receptor). Preferred agents for modulating apoptosis are agents that induce apoptosis, such as TNF or a TNF related ligand, particularly a ligand

TRAMP, Fas / CD95 ligand, TNFR-1 ligand, or TRAIL.

[0051] The IAP inhibitors of the present description are compounds having the general formula II:

<img file="PL2019671T3_D0001.tif" />

wherein:

A1 and A1 'are independently selected from the group consisting of hydrogen, optionally substituted alkyl and Z;

ή

A2 and A2 'are independently selected from the group consisting of hydrogen, optionally substituted alkyl and COR¹where A2 is not present when V is O and A2 'is not present when V' is O;

V and V 'are independently selected from the group consisting of N, CH and O; W and W 'are independently selected from the group consisting of

CH and N;

X and X 'are independently optionally substituted C 1-3 alkyl;

Y and Y 'are independently selected from the group consisting of CONR¹ , C (O) O, (CR¹R²) 1-3, where one or more CH2 groups can be replaced by O, S or NR¹, optionally substituted aryl and optionally substituted heteroaryl;

D and D 'are independently selected from the group consisting of optionally substituted alkenyl and (CR¹R²) n^5a(CR³R⁴) M;

EP 2 019 671 B1

J and J 'are independently selected from the group consisting of optionally substituted alkenyl and (CR¹R²) p^5b(CR³R⁴) Q;

T and T 'are independently selected from the group consisting of C = O, C = S, C = NR¹, S, O, NR¹, CR¹R², optionally substituted carbocycle, optionally substituted heterocycle, optionally substituted aryl and optionally substituted heteroaryl;

U and U 'are independently selected from the group consisting of hydrogen, NR¹R², OR¹ SR¹, optionally substituted alkyl and optionally substituted aryl; n, m, and qq are independently 0-5;

each R¹ is selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted carbocycle, optionally substituted heterocycle, optionally substituted aryl, optionally substituted heteroaryl and Z;

3 4 each R², R³ and R⁴ is independently selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted carbocycle, optionally substituted heterocycle, optionally substituted aryl and optionally substituted heteroaryl;

R^5a and R^5b are independently selected from the group consisting of C = O, C = S, C = NR¹, S, O, NR¹ and CR¹R²; and

Z is a linker covalently linking one of A1, Y, D, J, T and U with one of A1 ', Y', D ', J', T 'and U'; or pharmaceutically acceptable salts or prodrugs thereof.

[0052] In one example, Z binds D to U '. In another example, Z binds D to D '. In another example, Z binds U to U '. In the next example, before they are independently selected from 0-4 such that n + m is 3 or 4. In the next example, piq are independently selected from 0 and 1 such that p + q is 1. In the next example, before they are independently selected from 0-4 such that n + m is 3 or 4 and p and q are independently selected from 0 and 1 such that p + q is 1. In another example, T is C = O. In another example, U is NR¹R². In the next example, R^5b means CH2. In another example, Y is CONH, W is CH and V is N. In another example, A² and A.^2' are independently selected from the group consisting of hydrogen and optionally substituted alkyl.

[0053] IAP inhibitors may be compounds of formula III:

<img file="PL2019671T3_D0002.tif" />

in which A1, A2, V, W, X, Y, D, J, Z, A1 ', A2', V ', W, X', Y ', D', J 'and R² have the meanings as above; or pharmaceutically acceptable salts or prodrugs thereof.

[0054] In another preferred example, IAP inhibitors are compounds of formula IV:

EP 2 019 671 B1

<img file="PL2019671T3_D0003.tif" />

in which A1, A2, V, W, X, Y, D, J, Z, T, U, A1 ', A2', V ', W', X ', Y', D ', J', T ' and U 'are as above; or pharmaceutically acceptable salts or prodrugs thereof.

[0055] In another preferred example, IAP inhibitors are compounds of formula V:

<img file="PL2019671T3_D0004.tif" />

in which A1, A2, V, W, X, Y, D, J, Z, A1 ', A2', V ', W', X¹, Y ', D', J ', T', U 'and R² have the meanings as above; or pharmaceutically acceptable salts or prodrugs thereof.

[0056] In another preferred example, intermediates useful for the preparation of IAPs are compounds of formula XIII:

<img file="PL2019671T3_D0005.tif" />

wherein:

D "means (CR¹R²) n^5c- (CR³R⁴) M;

J is selected from the group consisting of optionally substituted alkenyl and (CR¹R²) p^5b- (CR³R⁴) Q;

T is selected from the group consisting of C = O, C = S, C = NR¹, S, O, NR¹, CR¹R², optionally substituted carbocycle, optionally substituted heterocycle, optionally substituted aryl and optionally substituted heteroaryl;

U is selected from the group consisting of hydrogen, NR¹R², OR¹, SR¹, optionally substituted alkyl and optionally substituted aryl;

n, m, and q are independently selected from 0-5;

each R¹, R², R³ and R⁴ is independently selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted carbocycle, optionally substituted heterocycle, optionally substituted aryl and optionally substituted heteroaryl;

EP 2 019 671 B1

R^5c is selected from the group consisting of C = O, C = S, C = NR¹, S, O, NR¹, CR^1aR^2a, NCOR⁸ and NCO2R⁸;

1a 2a

R^1a and R^2a are independently selected from the group consisting of hydrogen, hydroxyl, azido, optionally substituted alkyl, optionally substituted carbocycle, optionally substituted heterocycle, optionally substituted aryl and optionally substituted heteroaryl;

R^5b is selected from the group consisting of O, S, NR¹, CR¹R², C = O, C = S and C = NR¹;

R⁷ is selected from the group consisting of hydrogen, CO2R^7a and COCH (R^7b) N (R^7c) CO 2 R^7a;

7a

R^7a is selected from the group consisting of optionally substituted alkyl, optionally substituted carbocycle, optionally substituted heterocycle, optionally substituted aryl and optionally substituted heteroaryl;

R^7b is optionally substituted C1-3 alkyl;

7c

R^7c is selected from the group consisting of hydrogen and optionally substituted alkyl; and

R⁸ is selected from the group consisting of optionally substituted alkyl, optionally substituted carbocycle, optionally substituted heterocycle, optionally substituted aryl and optionally substituted heteroaryl.

7a 5c [0057] In another example, R^7a means t-butyl. In another example, n is 1, m is 2, R^5c

Ο Ο d ο * 1 ΓΊ stands for NCO₂R⁸ and R⁸ means benzyl. In the next example, R^5c means CR^1aR^2a, R^1a is selected from the group 2a consisting of hydroxyl, azido and optionally substituted heteroaryl and R^2a is hydrogen.

[0058] Useful alkyl groups include straight chain or branched C1-18 alkyl groups, especially methyl, ethyl, propyl, isopropyl, t-butyl, sec-butyl, 3-pentyl and 3-hexyl groups. The term "alkenyl" refers to a divalent alkyl radical containing 1, 2, 3 or 4 linked methylene groups, such as - (CH2) 4- for example.

[0059] Useful alkenyl groups include straight chain or branched C2-18 alkyl groups, especially ethenyl, propenyl, isopropenyl, butenyl, isobutenyl and hexenyl.

[0060] The term "alkenylene" as used herein refers to a divalent alkene derived radical, such as, for example, -CH2CH = CHCH2-.

[0061] Useful alkynyl groups are C2-18 alkynyl groups, especially ethynyl, propynyl, butynyl and 2-butynyl groups.

[0062] Useful cycloalkyl groups are C3-8 cycloalkyl. Typical cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, adamantyl and norbornyl.

[0063] Useful aryl groups include C6-14 aryl, especially phenyl, naphthyl, phenantrenyl, anthracenyl, indenyl, azulenyl, biphenyl, biphenylenyl and fluorenyl groups.

[0064] Useful heteroaryl groups include thienyl, benzo [b] thienyl, naphtho [2,3-b] thienyl, thiantrenyl, furyl, pyranyl, isobenzofuranyl, chromenyl, xanthenyl, phenoxantenyl, 2H-pyrrolyl, pyrrolyl, imidazolyl, triazolyl, pyrazilyl , pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolizinyl, isoindolyl, 3H-indolyl, indolyl, indazolyl, purinyl, 4H-quinolizinyl, isoquinolyl, quinolyl, phthalazinyl, naphthyridinyl, quinazalinyl, cynolinyl, phenyldyldynylterylidinyl) perimidinyl, phenanthrolinyl, phenynyl, isothiazolyl, phenothiazinyl, isoxazolyl, furazanyl, phenoxazinyl, 1,4-dihydroquinoxaline-2,3-dione, 7-aminoisocoumarin, pyrido [1,2-a] pyrimidin-4on, 1,2-benzisoxazole 3-yl, benzimidazolyl, 2-oxindolyl and 2-oxobenzimidazolyl. When the heteroaryl group contains a ring nitrogen atom, such a nitrogen atom may exist in the form of an N-oxide, e.g., pyridyl N-oxide, pyrazinyl N-oxide, pyrimidinyl N-oxide, and the like.

[0065] Optional substituents include one or more alkyl; halogen atoms; azido groups; haloalkyl; hydroxy; alkynyl; cycloalkyl; heteroalkyl; heteroalkinyli; aryls optionally substituted by one or more lower alkyl, halogen, haloalkyl or heteroaryl groups; aryloxy optionally substituted with one or more lower alkyl, haloalkyl or heteroaryl groups; aralkyl; heteroaryls optionally substituted with one or more lower alkyl, haloalkyl and aryl groups; heteroaryloxy optionally substituted with one or more lower alkyl, haloalkyl and aryl groups; alkoxy; alkylthio groups; arylthio groups; amide groups; amino groups; acyloksyli; arylacyloxy optionally substituted with one or more lower alkyl, haloalkyl and aryl groups; diphenylphosphinyloxy optionally substituted with one or more lower alkyl, halogen or haloalkyl groups; heterocycles optionally substituted with one or more lower alkyl, haloalkyl and aryl groups; heterocycloalkoxy optionally substituted with one or more lower alkyl, haloalkyl and aryl groups; partially unsaturated heterocycloalkyl optionally substituted with one or more lower alkyl, haloalkyl and aryl groups; partially unsaturated heterocycloalkyloxy optionally substituted with one or more lower alkyl, haloalkyl and aryl groups; and any covalent linker (see below).

[0066] Useful saturated or partially saturated carbocyclic groups are cycloalkyl groups as defined above as well as cycloalkenyl groups such as cyclopentenyl, cycloheptenyl and cyclooctenyl. Carbocyclic groups also include groups having fused optionally substituted aryl groups such as tetralin.

[0067] Useful halogen or halogen groups include fluorine, chlorine, bromine and iodine.

[0068] Useful alkylaryl and alkylheteroaryl groups include any of the above-mentioned C1-18 alkyl groups substituted with any of the above-mentioned C6-14 aryl or heteroaryl groups. Useful values ​​include benzyl, phenethyl and naphthylmethyl.

[0069] Useful haloalkyl groups include C 1-10 alkyl groups substituted with one or more fluorine, chlorine, bromine or iodine atoms, e.g., fluoromethyl, difluoromethyl, trifluoromethyl, pentafluoroethyl, 1,1-difluoroethyl, chloromethyl, chlorofluoromethyl and trichloromethyl and trichloromethyl and trichloromethyl groups.

[0070] Useful heteroalkyl groups include C1-10 alkyl groups containing one or more nitrogen, oxygen or sulfur atoms, e.g., -CH2CH2OCH3, -CH2OH, -CH2CH2NH2 and -CH2CH2NHCH3 groups.

[0071] Useful heteroalkynyl groups include C2-18 alkynyl groups containing one or more nitrogen, oxygen or sulfur atoms, e.g., -CH2OCH2CCH.

[0072] Useful alkoxy groups include oxygen substituted with one of the C 1-10 alkyl groups mentioned above.

[0073] Useful alkylthio groups include sulfur substituted with the one C1-10 alkyl group mentioned above. Also included are sulfoxides and sulfones of such alkylthio groups.

[0074] Useful amide groups include carbonylamide as well as any C1-6 acyl (alkanoyl) bonded with amine nitrogen, eg, acetamide, propionamide, butanoylamide, pentanoylamide, hexanoylamide as well as aryl substituted C2-6 substituted acyl groups.

[0075] Useful acyloxy groups are any C 1-6 acyl (alkanoyl) bonded to the oxy (-O-) group, e.g., formyloxy, acetoxy, propionyloxy, butanoyloxy, pentanoyloxy, hexanoyloxy and the like. [0076] Useful arylacyloxy groups include any of the aryl groups mentioned above substituted on any of the acyloxy groups mentioned above, e.g., 2,6-dichlorobenzoyloxy, 2,6-difluorobenzoyloxy and 2,6-di- (trifluoromethyl) -benzoyloxy groups.

[0077] Useful amino groups include -NH₂, -NHR¹¹, and -NR¹¹R¹¹where R¹¹ and R¹² are groups C_1-10alkyl or cycloalkyl as defined above.

[0078] Useful saturated or partially saturated heterocyclyl groups include tetrahydrofuranyl, pyranyl, piperidinyl, piperizinyl, pyrrolidinyl, imidazolidinyl, imidazolinyl, indolinyl, isoindolinyl, quinuclidinyl, morpholinyl, pyrrolidinyl, isochrinyl, isochrinyl, isochromyl

[0079] Useful arylene groups include C6-14 arylene, especially phenylene, naphthylene, phenantrenylene, anthracenylene, indenylene, azulenylene, biphenylene, biphenylene and fluorenylene groups.

[0080] Useful heteroarylene groups include disubstituted heteroaryl groups such as 2,5-thienylene, 2,4-imidazoylene and 1,3-triazolylene.

[0081] In the present specification, the groups and their optional substituents are selected to provide stable moieties and compounds.

[0082] Covalent linkers that can be used include any divalent covalent linker. The linker may be a continuous chain of 5 to 50 atoms. The linker typically has a length from about 5 angstroms to about 100 angstroms for standard bond lengths and angles. More preferably, the linker is from about 10 angstroms to about 50 angstroms. In some examples, the linker contains at least one aryl, heteroaryl, or heterocyclic moiety. In other examples, the linker is symmetrical. In other examples, the linker is asymmetrical. The linker may be any of many known homodifunctional and heterodifunctional linkers. See e.g. 805, 5 262 524, 5 258 498, 5 212 075, 5 165 923, 5 141 648.

[0083] In another example, the linker may include a group -COR^9a - or -R^9aCO- associated with any of A1, Y, D, J, T or U and any of A₁', Y', D ', J', T 'or U' where R^7a means O, S or NR^10a and R^10a is hydrogen or lower alkyl. In this example, the linker further includes a group related to R^9a- a first group and CO- a second group which may include an optionally substituted alkylene group, wherein any of the carbon atoms of the alkylene group may be substituted by one or more O, S, NR^10a, arylene and heteroarylene groups. Examples of such linkers include, without limitation:

<img file="PL2019671T3_D0006.tif" />

EP 2 019 671 B1

<img file="PL2019671T3_D0007.tif" />

EP 2 019 671 B1

<img file="PL2019671T3_D0008.tif" />

[0084] In another example, the linker may include a carbonyl group bonded to any of A1, Y, D, J, T or U and any of A1 ', Y', D ', J', T 'or U', and in addition includes an alkylene, polyalkylene or aralkyl glycol group bonded to carbonyl groups. Examples of such glycols include polyoxyethylene, polyoxypropylene, and block copolymers of polyoxyethylene and polyoxypropylene glycol, diethylene glycol, triethylene glycol, tetraethylene glycol, dipropylene glycol, thioethylene glycol and pentaethylene glycol, hexaethylene glycol, heptaethylene, and octaethylene, and octaethylene. Specific examples of these glycols include ethylene glycol; 1,2-propylene glycol; 1,3-propanediol; 2,4-dimethyl-2etyloheksano 1,3-diol; 2,2-dimethyl-1,3-propanediol; 2-ethyl-2-butyl-1,3-propanediol; 2-ethyl-2-izobutylo1,3-propanediol; 1,3-butanediol; 1,4-butanediol: 1,5-pentanediol; 1,6-hexanediol; 2,2-4-trimethyl-1,6-hexanediol; thiodiethanol; 1,2-cyclohexanedimethanol; 1,3-cyclohexanedimethanol; 1,4-cyclohexanedimethanol; 2,2,4,4-tetramethyl-1,3-cyclobutanediol; p-xylylenediol, 2,3-naphthalene diol and 2,7-naphthalene diol. Examples of diamine compounds include 1,3-bis- (2,4-diaminophenoxy) propane; 2,4-diamino5-metylofenetol; 2,4-diamino-5-metylofenoksyetanol; 2,4-diamino; 2,4-diaminophenol; 2,4diaminofenol; 2,4-diaminophenoxyethanol; 2,6-bis (2-hydroxyethoxy) -3,5-pyridinediamine; 2,6diaminopirydynę; 2,6-dimethoxy-3,5-pyridinediamine, 2-chloro-5-nitro-n-hydroxyethyl-p-phenylenediamine, 2-chloro-p-phenylenediamine, 2-aminomethyl-p-aminophenol and 4,5-diamino-1- methylpyrazole. Examples of amino-hydroxyl compounds include 2-amino-3-hydroxypyridine; 2-amino-3-nitrophenol; 2-amino-4hydroksyetyloaminoanizol; 2-amino-4-hydroxyethylaminoanisole sulfate; and 2-amino-6-chloro-4-nitrophenol. [0085] In another example, the linker may include oxygen or an amino group bonded to any of A1, Y, D, J, T or U and any of A1 ', Y', D ', J', T 'or U', and further include a diacid, thereby giving a diester, diamide or esteramide. Examples of such diacids include succinic acid, fumaric acid, adipic acid and the like.

[0086] In another example, the linker comprises a 1,2,3-triazol-4,5-ene group, which is introduced by cycloaddition of the propargyl group with an azido group.

[0087] The linker is used to join two Smac mimetic compounds into a divalent structure. Smac mimetic compounds combined with each other can be the same or different, and can be any compound

Mimetic Smac known for binding to IAP and inhibiting the interaction of IAP and caspases. In one example, Smac mimetics are conformational limited. In another example, Smac mimetics do not contain any naturally occurring amino acids. In another example, Smac mimetics do not contain any peptide bonds. Examples of known Smac mimetic compounds that are useful as substrates include the following:

[0088] WO 2005/069888 discloses Smac peptidomimetic compounds of formula VI:

<img file="PL2019671T3_D0009.tif" />

or a pharmaceutically acceptable salt or prodrug thereof, in which:

R1 is C1-2 alkyl or C1-2 haloalkyl;

R2 is branched or unbranched alkyl or cycloalkyl or substituted or unsubstituted aryl, alkylaryl, heteroaryl or alkylheteroaryl;

R3 is branched or unbranched alkyl or cycloalkyl or substituted or unsubstituted aryl, alkylaryl, heteroaryl or alkylheteroaryl;

Y is (CH2) 0-3, where one or more carbon atoms can be replaced by one or more heteroatoms selected from oxygen, sulfur, and nitrogen, and one or more hydrogen atoms in CH2 groups can be replaced by branched or unbranched alkyl or cyclic alkyl or substituted or unsubstituted aryl, alkylaryl, heteroaryl or alkylheteroaryl; and

Z is CONH, CH2O, NHCO, (CH2) 1-4, (CH2) 1-3CONH (CH2) 0-3, (CH2) 1-3S (CH2) 0-3, (CH2) 1-3NH (CH2) 0-3, (CH2) 1-3NHCO (CH2) 0-3, (CH2) 1-3NHSO2 (CH2) 0-3, (CH2) 1-3NHC (O) NH (CH2) 0-3, (CH2) 1-3NHC (S) NH (CH2) 0-3, or (CH2) 1-3NR '(CH2) 0-3, where R' is branched or unbranched alkyl or cycloalkyl or substituted or unsubstituted aryl, alkylaryl, heteroaryl or alkylheteroaryl .

[0089] WO 2005/069894 discloses Smac mimetic compounds of formula VII:

<img file="PL2019671T3_D0010.tif" />

or a pharmaceutically acceptable salt or prodrug thereof, in which:

R1 is C1-2 alkyl or C1-2 haloalkyl;

R2 is branched or unbranched alkyl or cycloalkyl or substituted or unsubstituted aryl, alkylaryl, heteroaryl or alkylheteroaryl;

X is CONH, CH2O, CH2NH, CH2S, or (CH2) 1-3;

Y1 is (CH2) 1-5, wherein one or more carbon atoms can be replaced by one or more heteroatoms selected from oxygen, sulfur and nitrogen, and one or more hydrogen atoms in CH2 groups can be replaced by branched or unbranched alkyl or cyclic alkyl or substituted or unsubstituted aryl, alkylaryl, heteroaryl or alkylheteroaryl;

Y2 is (CH2) 1-5, wherein one or more carbon atoms can be replaced by one or more heteroatoms selected from oxygen, sulfur and nitrogen, and one or more hydrocarbons.

Hydrogen in CH2 groups can be replaced by branched or unbranched alkyl or cyclic alkyl or substituted or unsubstituted aryl, alkylaryl, heteroaryl or alkylheteroaryl; and Z is CONH, CH2O, NHCO, (CH2) 1-4, (CH2) 1-3CONH (CH2) 0-3, (CH2) 1-3S (CH2) 0-3, (CH2) 1-3NH (CH2) ) 0-3, (CH2) 1-3NHCO (CH2) 0-3, (CH2) 1-3NHSO2 (CH2) 0-3, (CH2) 1-3NHC (O) NH (CH2) 0-3, (CH2 ) 1-3NHC (S) NH (CH2) 0-3, or (CH2) 1-3NR '(CH2) 0-3, wherein R' is branched or unbranched alkyl or cycloalkyl or substituted or unsubstituted aryl, alkylaryl, heteroaryl or alkylheteroaryl [0090] WO 2006/010118 discloses Smac mimetics compounds of formula VIII:

<img file="PL2019671T3_D0011.tif" />

or a pharmaceutically acceptable salt or prodrug thereof, in which:

A is NR1R2, or N⁺R1R2R3;

R1, R2, and R3 are independently hydrogen or optionally substituted C1-8 alkyl, C2-8 alkenyl, or C2-8 alkynyl in which one or more carbon atoms can be replaced by C = O, C = S, or a heteroatom selected from O, S, and N, and one or more hydrogen atoms in the CH, CH2 or CH3 groups may be replaced by a fluorine atom, branched or unbranched alkyl or cycloalkyl, optionally substituted aryl, alkylaryl, heteroaryl or alkylheteroaryl, or OR4 , SR4, or NR4R5;

R4 and R5 are independently hydrogen or optionally substituted C1-4 alkyl, C2-5 alkenyl or C2-5 alkynyl in which one or more carbon atoms can be replaced by a heteroatom selected from O, S, and N, or optionally substituted aryl, alkylaryl, heteroaryl or alkylheteroaryl; or any two of R1, R2, and R3 taken together with the nitrogen atom to which they are attached form a heterocyclic group in which one or more carbon atoms can be replaced by C = O, C = S, or a heteroatom selected from O , S, and N, provided that the heteroatom separates at least two carbon atoms from the nitrogen atom;

B is optionally substituted C 1-4 alkyl, C 2-4 alkenyl or C 2-4 alkynyl in which one or more hydrogen atoms can be replaced by a fluorine atom;

U is CONH, C (O) O, C (S) O, C (S) NH, C (NH) NH or (CH2) 1-5, where one or more carbon atoms can be replaced by a heteroatom selected from O , S, and N;

V and W are independently (CH2) 1-5, wherein one or more carbon atoms can be replaced by C = O, C = S, or a heteroatom selected from O, S, and N, and one or more hydrogen atoms in CH2 groups it may be replaced by branched or unbranched alkyl or cycloalkyl, optionally substituted aryl, alkylaryl, heteroaryl or alkylheteroaryl, or OR4, SR4, or NR4R5;

X is optionally substituted C1-18 alkyl, C2-18 alkenyl, C2-18 alkynyl, aryl or heteroaryl in which one or more carbon atoms can be replaced by C = O, C = S, or a heteroatom selected from O, S , and N, and one or more hydrogen atoms in the groups CH, CH2 or CH3 may be replaced by branched or unbranched alkyl or cycloalkyl, optionally substituted aryl, alkylaryl, heteroaryl or alkylheteroaryl, or OR4, SR4, or NR4R5;

Y is CH or N;

Z is CH2, C = O, C = S, CHSR, CHOR or CHNR; and

R is hydrogen or optionally substituted C 1-4 alkyl, C 2-4 alkenyl or C 2-4 alkynyl.

EP 2 019 671 B1 [0091] Published US Application No. 2005/0234042 discloses compounds corresponding to Formula IX:

<img file="PL2019671T3_D0012.tif" />

wherein R1 is H; C1-C4 alkyl; C1-C4 alkenyl; C1-C4 alkynyl or C3-C10 cycloalkyl which are unsubstituted or substituted;

R2 is H; C1-C4 alkyl; C1-C4 alkenyl; C1-C4 alkynyl or C3-C10 cycloalkyl which are unsubstituted or substituted;

R3 is H; CF3; -C2F5; C1-C4 alkyl; C1-C4 alkenyl; C1-C4 alkynyl; -CH2-Z or R2 and R3 together with the nitrogen form a het ring;

Z is H; OH; F; Cl; CH3; CF3; -CH2Cl; -CH2F or -CH2OH;

R4 is C1-C16 straight or branched alkyl; C1-C16 alkenyl; C1-C16 alkynyl; or -C3-C10 cycloalkyl; - (CH2) 16-Z1; - (CH2) 0-6-aryl; and - (CH2) 0-6-het; wherein alkyl, cycloalkyl and phenyl are unsubstituted or substituted;

Z is -N (R8) -C (O) -C1-C10 alkyl; -N (R8) -C (O) - (CH2) 1-6-C3-C7 cycloalkyl; -N (R8) -C (O) - (CH2) 0-6-phenyl; -N (R8) C (O) - (CH2) 1-6-het; -C (O) -N (R9) (R10); -C (O) -O-C1-C10 alkyl; -C (O) -O- (CH2) 1-6-C3-C7 cycloalkyl; -C (O) -O- (CH2) 06-phenyl; -C (O) -O- (CH2) 1-6-het; -OC (O) -C1-C10 alkyl; -OC (O) - (CH2) 1-6-C3-C7 cycloalkyl; -OC (O) - (CH2) 0-6-phenyl; OC (O) - (CH2) 1-6-het; wherein alkyl, cycloalkyl and phenyl are unsubstituted or substituted; het is a 5-7 membered heterocyclic ring containing 1-4 heteroatoms selected from N, O and S, or an 8-12 membered fused ring system comprising at least one 5-7 membered heterocyclic ring containing 1, 2 or 3 heteroatoms selected from N, O and S, which heterocyclic ring or fused ring system is unsubstituted or substituted on carbon or nitrogen;

R8 is H; CH3; CF3; -CH2OH or -CH2Cl;

R9 and R10 are independently H; C1-C4 alkyl; C3-C7 cycloalkyl; - (CH2) 1-6-C3-C7 cycloalkyl; - (CH2) 0-6-phenyl; wherein alkyl, cycloalkyl and phenyl are unsubstituted or substituted, or R9 and R10 together with the nitrogen form a het;

R5 is H; C1-C10-alkyl; aryl; phenyl; C3-C7 cycloalkyl; - (CH2) 1-6-C3-C7 cycloalkyl; - C1-C10 alkyl aryl; - (CH2) 06-C3-C7 cycloalkyl- (CH2) 0-6-phenyl; - (CH2) 0-4CH - ((CH2) 1-4-phenyl) 2; - (CH2) 0-6-CH (phenyl) 2; -indanyl; -C (O) -C1-C10 alkyl; -C (O) - (CH2) 1-6-C3-C7-cycloalkyl; -C (O) - (CH7) 0-6-phenyl; - (CH2) 0-6-C (O) -phenyl; - (CH2) 0-6-het; -C (O) - (CH2) 1-6het; or R5 is an amino acid residue where the alkyl, cycloalkyl, phenyl and aryl substituents are unsubstituted or substituted;

U is as shown in the structure X:

<img file="PL2019671T3_D0013.tif" />

in which n = 0-5;

X is -CH or N;

Ra and Rb are independently O, S or N or C 0-8 alkyl, where one or more carbon atoms in the alkyl chain may be replaced with a heteroatom selected from O, S or N, and wherein alkyl may be unsubstituted or substituted;

EP 2 019 671 B1

Rd is selected from: (a) -Re-Q- (Rf) p (Rg) q; or (b) Ar1-D-Ar2;

Rc is H or Rc and Rd may together form cycloalkyl or het; where if Rd and Rc form cycloalkyl or het, R5 is bonded to the resulting ring on C or N; piq are independently 0 or 1;

Re is C1-8 alkyl or alkylidene, and Re which may be unsubstituted or substituted;

Q is N, O, S, S (O), or S (O) 2;

Ar1 and Ar2 are substituted or unsubstituted aryl or het;

Rf and Rg are independently H; -C1-C10 alkyl; C1-C10 alkylaryl; OH; -O-C1-C10 alkyl; - (CH2) 0-6-C3-C7 cycloalkyl; -O- (CH2) 0-6-aryl; phenyl; aryl; phenyl-phenyl; - (CH2) 1-6-het; -O- (CH2) 1-6-het; -OR11; -C (O) -R11; -C (O) N (R₁₁) (R₁₂); -N (R₁₁) (R₁₂); - SR₁₁; -S (O) -R₁₁; -S (O)₂-R₁₁; -S (O)₂-NR₁₁R₁₂; -NR₁₁-S (O)₂-R₁₂; SC₁-C₁₀ alkyl; aryl-C1-C4 alkyl; het-C1-C4-alkyl where alkyl, cycloalkyl, het and aryl are unsubstituted or substituted; -SO2-C1-C2 alkyl; -SO2-C1-C2 alkylphenyl; -O-C1-C4 alkyl; or Rg and Rf form a ring selected from het or aryl;

D is -CO-; -C (O) -C1-7 alkylene or arylene; -CF2-; -ABOUT-; -S (O) r where r is 0-2; 1,3-dioxolane; or C1-7 alkyl-OH; wherein alkyl, alkylene or arylene may be unsubstituted or substituted by one or more halogen, OH, -O-C1-C6 alkyl, -S-C1-C6 alkyl or -CF3; or D is -N (Rh) wherein Rh is H; C1-7 alkyl (unsubstituted or substituted); aryl; -O (C1-7 cycloalkyl) (unsubstituted or substituted); C (O) -C1-C10 alkyl; C (O) -C0-C10 alkyl aryl; CO-C1-C10 alkyl; CO-C0-C10 alkyl-aryl or SO2-C1-C10-alkyl; SO2 (C0-C10-alkylaryl);

R6, R7, R'6 and R'7 are independently H; -C1-C10 alkyl; -C1-C10 alkoxy; aryl-C1-C10 alkoxy; OH; -O-C1-C10 alkyl; - (CH2) 0-6-C3-C7 cycloalkyl; -O- (CH2) 0-6-aryl; phenyl; - (CH2) 1-6-het; -O- (CH2) 1-6-het; -OR11; -C (O) -R¹¹; -C (O) N (R11) (R12); - N (R11) (R12); -S-R11; -S (O) -R11; -S (O) 2-R11; -S (O) 2-NR11R12; -NR 11 -S (O) 2-R12; wherein alkyl, cycloalkyl and aryl are unsubstituted or substituted; and R6, R7, R'6 and R'7 can be combined to form a ring system;

R11 and R12 are independently H; C1-C10 alkyl; - (CH2) 0-6-C3-C7 cycloalkyl; - (CH2) 0-6- (CH) 0-1 (aryl) 1-2; -C (O) -C1C10 alkyl; -C (O) - (CH2) 1-6-C3-C7 cycloalkyl; -C (O) -O- (CH2) 0-6-aryl; -C (O) - (CH2) 0-6-O-fluorenyl; -C (O) -NH- (CH2) 0-6aryl; -C (O) - (CH2) 0-6-aryl; -C (O) - (CH2) 1-6-het; -C (S) -C1-C10alkyl; -C (S) - (CH2) 1-6-C3-C7 cycloalkyl; -C (S) -O- (CH2) 06-aryl; -C (S) - (CH2) 1-6-O-fluorenyl; -C (S) -NH- (CH2) 0-6-aryl; -C (S) - (CH2) 0-6-aryl; -C (S) - (CH2) 1-6-het; wherein alkyl, cycloalkyl and aryl are unsubstituted or substituted; or R11 and R12 are a substituent that facilitates transport of the molecule across a cell membrane; or R11 and R12 together with the nitrogen form a het; wherein the R11 and R12 alkyl substituents may be unsubstituted or substituted with one or more substituents selected from C1-C10 alkyl, halogen, OH, -O-C1-C6 alkyl, - S-C1-C6 alkyl or -CF3; substituted cycloalkyl substituents R11 and R12 are substituted with one or more substituents selected from C1-C10 alkene; C1-C6 alkyl; halogen; OH; -O-C1-C6 alkyl; -S-C1-C6 alkyl or -CF3; and substituted phenyl or aryl R11 and R12 are substituted with one or more substituents selected from halogen; hydroxy; C1-C4 alkyl; C1-C4 alkoxy; nitro group; CN; -OC (O) -C1-C4 alkyl and -C (O) -O-C1-C4-aryl, or pharmaceutically acceptable salts thereof.

[0092] US Published Application No. 2005/0261203 discloses compounds of formula XI:

EP 2 019 671 B1

<img file="PL2019671T3_D0014.tif" />

wherein X1 and X2 are independently O or S;

L is a bond, -C (X3) -, -C (X3) NR12 or -C (X3) O- where X3 is O or S and R12 is H or R1;

R1 is alkyl, carbocycle, carbocyclic substituted alkyl, heterocycle or heterocyclic substituted alkyl, each of which is optionally substituted with halogen, hydroxy, mercapto, carboxy, alkyl, haloalkyl, alkoxy, alkylsulfonyl, amino, nitro, aryl and heteroaryl;

R2 is alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocycle or heterocyclylalkyl;

R3 is H or alkyl;

R4 and R4 'are independently H, alkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, heteroaryl, or heteroaralkyl, each of which is optionally substituted with halogen, hydroxy, mercapto, carboxy, alkyl, alkoxy, amino and nitro;

R5 and R5 'are independently H or alkyl;

R6 is H or alkyl;

and their salts and solvates.

<img file="PL2019671T3_D0015.tif" />

where X1, X2 and X3 are independently O or S;

Y is (CHR7) n, O or S; where n is 1 or 2 and R7 is H, halogen, alkyl, aryl, aralkyl, amino, arylamino, alkylamino, aralkylamino, alkoxy, aryloxy or aralkyloxy;

A is a 5-membered heterocycle consisting of 1 to 4 heteroatoms optionally substituted with amino, hydroxy, mercapto, halogen, carboxy, amidino, guanidino, alkyl, alkoxy, aryl, aryloxy, acyl, acyloxy, acylamino, alkoxycarbonylamino, cycloalkyl, alkylthio, alkylsulfinyl, alkylsulfonyl, aminosulfonyl, alkylaminosulfonyl, alkylsulfonylamino or heterocycle; wherein each substitution with alkyl, alkoxy, aryl, aryloxy, acyl, acyloxy, acylamino, cycloalkyl and heterocycle is optionally substituted with hydroxyl, halogen, mercapto, carboxyl, alkyl, alkoxy, haloalkyl, amino, nitro, cyano, cycloalkyl , aryl or heterocycle;

R1 is H or R1 and R2 together form a 5-8 membered ring;

R2 is alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocycle or heterocyclylalkyl; each possibly

Substituted with hydroxyl, mercapto, halogen, amino, carboxy, alkyl, haloalkyl, alkoxy or alkylthio;

R3 is H or alkyl;

R4 and R4 'are independently H, hydroxy, amino, alkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, heteroaryl, or heteroarylalkyl, where each alkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, heteroaryl and heteroarylalkyl is optionally substituted with halogen, hydroxy, mercapto, carboxy, alkyl, alkoxy, amino and nitro;

R5 and R5 'are independently H or alkyl;

R6 and R6 'are independently H, alkyl, aryl or aralkyl;

and their salts and solvates [0094] Certain compounds of the present description may exist as stereoisomers, including optical isomers. All stereoisomers are included as pure individual preparations of stereoisomers and their enriched preparations, and racemic mixtures of such stereoisomers, as well as individual enantiomers that can be separated according to methods well known to those skilled in the art.

[0095] The compound of formula II may be selected from the group consisting of:

<img file="PL2019671T3_D0016.tif" />

<img file="PL2019671T3_D0017.tif" />

EP 2 019 671 B1

<img file="PL2019671T3_D0018.tif" />

<img file="PL2019671T3_D0019.tif" />

EP 2 019 671 B1

<img file="PL2019671T3_D0020.tif" />

<img file="PL2019671T3_D0021.tif" />

<img file="PL2019671T3_D0022.tif" />

EP 2 019 671 B1

<img file="PL2019671T3_D0023.tif" />

EP 2 019 671 B1

<img file="PL2019671T3_D0024.tif" />

or their free base or other pharmaceutically acceptable salt thereof.

[0096] The compounds may be prepared using methods known to those skilled in the art. Specifically, compounds of formula II can be prepared as illustrated by the exemplary reactions in the Examples.

[0097] An important aspect is that the compounds of formula II induce apoptosis and also enhance the induction of apoptosis in response to signals that induce apoptosis. Thus, it is contemplated that these compounds sensitize cells to agents that cause apoptosis, including cells resistant to such agents. IAP inhibitors can be used to induce apoptosis in any disorder that can be treated, alleviated or prevented by inducing apoptosis. Thus, the present description provides compositions and methods for targeting animals characterized as overexpressing the IAP protein. In some examples, cells (e.g., cancer cells) exhibit elevated levels of IAP protein expression compared to non-pathological samples (e.g., non-cancerous cells). In other examples, cells in action exhibit elevated levels of IAP protein expression by performing an apoptosis program and dying in response to inhibition of an effective amount of a compound of Formula I where the response occurs, at least in part, due to the dependence of the survival of such cells on the action of the IAP protein.

[0098] The present disclosure also relates to the modulation of a condition associated with apoptosis that is associated with one or more agents that modulate apoptosis. Examples of agents that modulate apoptosis include, but are not limited to, Fas / CD95, TRAMP, TNF RI, DR1, DR2, DR3, DR4, DR5, DR6, FADD, RIP, TNFα, Fas ligand, TRAIL, antibodies to TRAIL-R1 or TRAIL-R2, Bcl-2, p53, BAX, BAD, Akt, CAD, PI3 kinase, PP1, and caspase proteins. Other measures involved in the initiation, decision and degradation phases of apoptosis are also included. Examples of agents that modulate apoptosis include agents whose activity, presence or change in concentration can modulate apoptosis in a subject. Preferred agents for modulating apoptosis are apoptosis-inducing agents such as TNF or TNF-related ligand, particularly TRAMP ligand, Fas / CD95 ligand, TNFR-1 ligand, or TRAIL.

[0099] The compositions and methods of the present description may be used to treat diseased cells, tissues, organs or pathological conditions and / or disease states in an animal (e.g., a mammalian subject including, but not limited to, humans and veterinary animals). In this regard, various diseases and pathologies are amenable to treatment or prevention using the present methods and compositions. A non-limiting example list of these diseases and conditions includes, but is not limited to, breast cancer, prostate cancer, lymphoma, skin cancer, pancreatic cancer, colon cancer, melanoma, malignant melanoma, ovarian cancer, brain cancer, primary brain cancer, head and neck cancer, glioblastoma, glioblastoma multiforme, liver cancer, bladder cancer, non-small cell lung cancer, head or neck cancer, breast cancer, ovarian cancer, lung cancer, small cell lung cancer, Wilms tumor, cervical cancer, testicular cancer, bladder cancer, pancreatic cancer, stomach cancer, colon cancer, prostate cancer, urogenital cancer, thyroid cancer, esophageal cancer, myeloma, multiple myeloma, adrenal cancer, kidney cell cancer, endometrial cancer, adrenocortical carcinoma, malignant pancreatic islet cell adenoma, malignant carcinoid, malignant chorionic epithelium, mycosis fungoides, malignant hypercalcaemia, hyperplasia of the cervix, leukemia, acute lymphocytic leukemia, chronic lymphocytic leukemia, acute myeloid leukemia, chronic myelogenous leukemia, chronic granulocytic leukemia, acute granulocytic leukemia, hair cell leukemia, neuroblastoma, rhabdomyosarcoma, angiospermoma, Kaposoma Hodgkin, non-Hodgkin's lymphoma, soft tissue sarcoma, osteogenic sarcoma, primary macroglobulinemia and retinoblastoma, and similar, T-cell and B-cell mediated autoimmune diseases; inflammatory diseases; infections; hyperplasia diseases; AIDS; degenerative conditions, diseases

Vascular, and the like. In certain embodiments, the cancer cells treated are metastases. In other embodiments, the treated cancer cells are resistant to anti-cancer agents.

[0100] Infections suitable for treatment with the compositions and methods of the present description include, but are not limited to, infections caused by viruses, bacteria, fungi, mycoplasma, prions and the like.

[0101] The present description discloses methods of administering an effective amount of a compound of formula I and at least one additional therapeutic agent (including, but not limited to, chemotherapeutic anti-cancer agents, apoptosis-modulating agents, antimicrobials, anti-viral agents, anti-fungal agents and anti-inflammatory agents ) and / or therapeutic technique (e.g., surgery and / or radiation therapy).

[0102] A lot of suitable anti-cancer agents are anticipated for use in the methods of the present description. Indeed, the present description provides, but is not limited to, the administration of numerous anti-cancer agents such as: apoptosis-inducing agents; polynucleotides (e.g., antisense, ribozymes, siRNA); polypeptides (e.g., enzymes and antibodies); biological mimetics (e.g., gossypol or BH3 mimetics); agents that bind (e.g., oligomerizing or complexing) to Bcl-2 family proteins such as Bax; alkaloids; alkylating agents; anti-cancer antibiotics; antimetabolites; hormones; platinum compounds; monoclonal or polyclonal antibodies (e.g., antibodies conjugated to anti-cancer drugs, toxins, defensins), toxins; radionuclides; biological response modifiers (e.g., interferons (e.g., IFNα) and interleukins (e.g., IL-2)); adoptive immunotherapy; hematopoietic growth factors; agents that induce tumor cell differentiation (e.g., fully trans retinoic acid); gene therapy reagents (e.g., antisense therapy reagents and nucleotides); cancer vaccines; angiogenesis inhibitors; proteasome inhibitors; NF-KB modulators; anti-CDK compounds; HDAC inhibitors; and the like. Numerous other examples of chemotherapeutic compounds and anti-cancer therapies suitable for co-administration with the disclosed compounds are known to those skilled in the art.

[0103] Anti-cancer agents may include agents that induce or stimulate apoptosis. Agents that induce apoptosis include, but are not limited to, radiation (e.g., X-rays, gamma rays, UV); tumor necrosis factor (TNF) related factors (e.g., TNF family receptor proteins, TNF family ligands, TRAIL, TRAIL-R1 or TRAIL-R2 antibodies); kinase inhibitors (e.g., epidermal growth factor receptor (EGFR) inhibitor, vascular growth factor receptor (VGFR) kinase, fibroblast growth factor receptor (FGFR) kinase inhibitor, platelet derived growth factor receptor (PDGFR) kinase inhibitor, and inhibitors Bcr-Abl kinases (such as GLEEVEC)); antisense molecules; antibodies (e.g., HERCEPTIN, RITUXAN, ZEVALIN and AVASTIN); anti-estrogens (e.g., raloxifene and tamoxifen); anti-androgens (e.g., flutamide, bicalutamide, finasteride, aminoglutetamide, ketoconazole and corticosteroids); cyclooxygenase 2 (COX-2) inhibitors (e.g., celecoxib, meloxicam, NS-398, and nonsteroidal anti-inflammatory drugs (NSAIDs)); anti-inflammatory drugs (e.g., butazolidine, DECADRON, DELTASONE, dexamethasone, dexamethasone intensol, DEXONE, HEXADROL, hydroxychloroquine, METICORTEN, ORADEXON, ORASONE, oxyfenbutazone, PEDIAPRED, phenylbutazone, predisonol, prednisone, prednisone, and chemotherapeutic drugs for cancer (e.g., irinotecan (CAMPTOSAR), CPT-11, fludarabine (FLUDARA), dacarbazine (DTIC), dexamethasone, mitoxantrone, MYLOTARG, VP-16, cisplatin, carboplatin, oxaliplatin, 5-FUnin gemcitabine, bortezomib, gefitinib, bevacizumab, TAXOTERE or TAXOL); cell signaling molecules; ceramides and cytokines; staurosporin, and the like.

[0104] The compositions and methods of the present description provide a compound of formula II and at least one antiproliferative or anti-tumor agent selected from alkylating agents, anti-metabolites and natural products (eg, herbs and other compounds derived from plants and / or animals). [0105] Alkylating agents suitable for use in the present compositions and methods include, but are not limited to: 1) nitrogen mustards (e.g., mechloretamine, cyclophosphamide, ifosfamide, melphalan (Lsarkolysin); and chlorambucil); 2) ethyleneimine and methylmelamine (e.g., hexamethylmelamine and thiotepa); 3) alkyl sulfonates (e.g., busulfan); 4) nitrosoureas (e.g., carmustine (BCNU); lomustine (CCNU); semustine (methyl-CCNU); and streptozocin (streptozotocin)); and 5) triazenes (e.g., dacarbazine (DTIC; dimethyltriazenoimido-azolecarboxamide)).

[0106] For example, antimetabolites suitable for use in the present compositions and methods include, but are not limited to: 1) folic acid analogues (e.g., methotrexate (ametopterin)); 2) pyrimidine analogues (e.g., fluorouracil (5-fluorouracil; 5-FU), floxuridine (fluorodeoxyuridine; FudR), and cytarabine (cytosine arabinoside)); and 3) purine analogues (e.g., mercaptopurine (6-mercaptopurine; 6-MP), thioguanine (6-thioguanine; TG), and pentostatin (2'-deoxycoformycin)).

[0107] In further examples, chemotherapeutic agents suitable for use in the compositions and methods of the present description include, but are not limited to: 1) vinca alkaloids (e.g., vinblastine (VLB), vincristine); 2) epipodophyllotoxins (e.g., etoposide and teniposide); 3) antibiotics (e.g., dactinomycin (actinomycin D), daunorubicin (daunomycin; rubidomycin), doxorubicin, bleomycin, fileamycin (mitramycin), and mitomycin (mitomycin C)); 4) enzymes (e.g., L-asparaginase); 5) biological response modifiers (e.g., interferon-alpha); 6) platinum coordination complexes (e.g., cisplatin (cisDDP) and carboplatin); 7) anthracenedione (e.g., mitoxantrone); 8) substituted ureas (e.g., hydroxyurea); 9) methylhydrazine derivatives (e.g., procarbazine (N-methylhydrazine; MIH)); 10) corticrenal suppressants (e.g., mitotane (o, p'-DDD) and aminoglutethimide); 11) adrenocorticosteroids (e.g., prednisone); 12) progestins (e.g., hydroxyprogesterone caproate, medroxyprogesterone acetate, and megestrol acetate); 13) estrogens (e.g., diethylstilbestrol and ethinyl estradiol); 14) anti-estrogens (e.g., tamoxifen); 15) androgens (e.g., testosterone propionate and fluoxymesterone); 16) antiandrogens (e.g., flutamide): and 17) gonadotropin-releasing hormone analogues (e.g., leuprolide).

[0108] Any oncolytic agent routinely used in the context of cancer therapy finds use in the compositions and methods of the present description. For example, the US Food and Drug Administration maintains a collection of oncolytic agents authorized for use in the United States. USFDA international counterparts maintain similar collections. Table 1 provides a list of exemplary anti-cancer agents approved for use in the USA. Those skilled in the art will recognize that the "product labels" required for all US approved chemotherapeutic agents describe recommended indications, dosage information, toxicity data, and the like for exemplary agents.

Table 1

<td>aldesleukin (des-alanyl-1, serine-125 human interleukin-2)</td><td>Proleukin</td><td>Chiron Corp., Emeryville, CA</td>

<td>alemtuzumab (anti-CD52 IgG1x antibody)</td><td>Campath</td><td>Millennium and ILEX Partners, LP, Cambridge, MA</td>

EP 2 019 671 B1

<td>alitretinoin (9-cis-retinoic acid)</td><td>Panretin</td><td>Ligand Pharmaceuticals, Inc., San Diego CA</td>

<td>allopurinol (1,5-dihydro-4H-pyrazolo [3,4-d] pyrimidin-4-one, monosodium salt)</td><td>Zyloprim</td><td>GlaxoSmithKline, Research Triangle Park, NC</td>

<td>altretamine (N, N, N ', N', N ", N" -heksametylo-1,3,5-triazine-2,4,6-triamine)</td><td>Hexalen</td><td>US Bioscience, West Conshohocken, PA</td>

<td>amifostine (2 - [(3-aminopropyl) amino] -ethanethiol dihydrogen phosphate, (ester))</td><td>Ethyol</td><td>US Bioscience</td>

<td>anastrozole (a, a, a ', a'-tetramethyl-5- (1H-1,2,4-triazol-1-ylmethyl) -1,3-benzenediacetonitrile)</td><td>Arimidex</td><td>AstraZeneca Pharmaceuticals, LP, Wilmington, DE</td>

<td>Arsenic trioxide</td><td>Trisenox</td><td>Cell Therapeutic, Inc., Seattle, WA</td>

<td>asparaginase (L-aspartic amidohydrolase, type EC-2)</td><td>Elspar</td><td>Merck &amp; Co., Inc., Whitehouse Station, NJ</td>

<td>Live BCG (lyophilized preparation of the weakened strain of Mycobacterium bovis (Bacillus Calmette-Gukin [BCG], Montreal sub-strain)</td><td>TICE BCG</td><td>Teknika Organon, Corp., Durham, NC</td>

<td>Bexarotene capsules (4- [1- (5,6,7,8-tetrahydro-3,5,5,8,8-pentamethyl-2-naphthalenyl) ethenyl] benzoic acid)</td><td>Targretin</td><td>Ligand Pharmaceuticals</td>

<td>bexarotene gel</td><td>Targretin</td><td>Ligand Pharmaceuticals</td>

<td>bleomycin (cytotoxic glycopeptide antibiotics produced by Streptomyces verticillus; bleomycin A2 and bleomycin R2)</td><td>Blenoksane</td><td>Bristol-Myers Squibb Co., NY, NY</td>

<td>capecitabine (5'-deoxy-5-fluoro-N - [(pentyloxy) carbonyl] cytidine)</td><td>Xeloda</td><td>Roche</td>

<td>carboplatin (diamino [1,1-cyclobutanedicarboxylate (2 -) - 0 '] - platinum (SP4-2))</td><td>Paraplatin</td><td>Bristol-Myers Squibb</td>

<td>carmustine (1,3-bis (2-chloroethyl) -1-nitrosourea)</td><td>BCNU, BiCNU</td><td>Bristol-Myers Squibb</td>

<td>Carmustine implant with polifeprosan 20</td><td>Gliadel Wafer</td><td>Guilford Pharmaceuticals, Inc., Baltimore, MD</td>

<td>Celecoksyb (as 4- [5- (4-methylphenyl) -3- (trifluoromethyl) -1H-pyrazol-1-yl] benzenesulfonamide)</td><td>Celebrex</td><td>Searle Pharmaceuticals, England</td>

<td>chlorambucil (4- [bis (2-chloroethyl) amino] benzenebutanoic acid)</td><td>Leukeran</td><td>GlaxoSmithKline</td>

<td>cisplatin (PtCl₂H₆N₂)</td><td>Platinol</td><td>Bristol-Myers Squibb</td>

<td>cladribine (2-chloro-2'-deoxy-bD-adenosine)</td><td>Leustatin, 2-CdA</td><td>RW Johnson Pharmaceutical Research Institute, Raritan, NJ</td>

EP 2 019 671 B1

<td>cyclophosphamide (2- [bis (2-chloroethyl) amino] tetrahydro-2H13.2-oxazaphosphite 2-oxide monohydrate)</td><td>Cytoxan, Neosar</td><td>Bristol-Myers Squibb</td>

<td>cytarabine (1-bD-arabinofuranosylcytosine, C9H13N3O5)</td><td>Cytosar-U</td><td>Pharmacia &amp; Upjohn Company</td>

<td>liposomal cytarabine</td><td>DepoCyte</td><td>Skye Pharmaceuticals, Inc., San Diego, CA</td>

<td>dacarbazine (5- (3,3-dimethyl-1-triazeno) -imidazole-4-carboxamide (DTIC))</td><td>DTIC-Dome</td><td>Bayer AG, Leverkusen, Germany</td>

<td>Dactinomycin, actinomycin D (actinomycin produced by Streptomyces parvullus, C62H86N12O16)</td><td>Cosmegen</td><td>Merck</td>

<td>Darbepoetin alfa (recombinant peptide)</td><td>Aranesp</td><td>Amgen, Inc., Thousand Oaks, CA</td>

<td>liposomal daunorubicin ((8S-cis) -8-acetyl-10 - [(3-amino-2,3,6-trideoxy-L-lixo-hexopyranosyl) oxy] hydrochloride] -7,8,9,10-tetrahydro-6.8 , 11trihydroksy-1-methoxy-5,12-naftacenodionu)</td><td>Danuoxome</td><td>Nexstar Pharmaceuticals, Inc., Boulder, CO</td>

<td>Daunorubicin HCl, daunomycin ((1 S, 3 S) -3-acetyl-1,2,3,4,6,11-hexahydro-3,5,12-trihydroxy-10-methoxy-6,11-dioxo-1-naphthacenyl-3-amino-hydrochloride 2,3,6-trideoksy- (alpha) -L-lyxo-heksopiranozydu)</td><td>Cerubidine</td><td>Wyeth Ayerst, Madison, NJ</td>

<td>Denileukin diftytoks (recombinant peptide)</td><td>Ontak</td><td>Seragen, Inc., Hopkinton, MA</td>

<td>dexrazoxane ((S) -4,4 '- (1-methyl-1,2-ethanediyl) bis-2,6-piperazinedione)</td><td>Zinecard</td><td>Pharmacia &amp; Upjohn Company</td>

<td>docetaxel (N-tert-butyl ester trihydrate, (2R, 3S) -N-carboxy-3-phenylisoserine 13-ester with 4-2-benzoate acetate 5b-20-epoxy12a, 4.7b, 10b, 13a-hexahydroxytax-11-en- 9-one)</td><td>Taxotere</td><td>Aventis Pharmaceuticals, Inc., Bridgewater, NJ</td>

<td>Doxorubicin HCl</td><td>Adriamycin, Ru-</td><td>Pharmacia &amp; Upjohn</td>

<td>(8S, 10S) -10 - [(3-amino-2,3,6-trideoxy-α-L-loxohexopyranosyl) oxy] -8-glycolyl-7,8,9,10-tetrahydro-6,8,11 trihydroxy- 1-methoxy-5,12-naftacenodionu)</td><td>bex</td><td>Company</td>

<td>doxorubicin</td><td>Adriamycin PFS Intravenous injection</td><td>Pharmacia &amp; Upjohn Company</td>

<td>liposomal doxorubicin</td><td>Doxil</td><td>Sequus Pharmaceuticals, Inc., Menlo park, CA</td>

<td>Dromostanolone propionate (17b-hydroxy-2a-methyl-5a-androstan-3-one propionate)</td><td>dromostanolone</td><td>Eli Lilly &amp; Company, Indianapolis, IN</td>

<td>Dromostanolone propionate</td><td>Masterone injection</td><td>Syntex, Corp., Palo Alto, CA</td>

<td>Elliott's solution B</td><td>Elliott's B Solution</td><td>Orphan Medical, Inc.</td>

EP 2 019 671 B1

<td>epirubicin ((8S-cis) hydrochloride -10 - [(3-amino-2,3,6-trideoxy-α-Larabino-hexopyranosyl) oxy] -7,8,9,10-tetrahydro-6,8,11 trihydroxy-8 - (hydroxyacetyl) -1-methoxy-5,12naftacenodionu)</td><td>Ellence</td><td>Pharmacia &amp; Upjohn Company</td>

<td>Epoetin alfa (recombinant peptide)</td><td>Epogen</td><td>Amgen, Inc.</td>

<td>estramustine (3- (bis (2-chloroethyl) carbamate) disodium salt monohydrate 17- (dihydrogen phosphate) (17 (beta)) - 1,3,5 (10) -triene-3,17-diol ester, or disodium salt monohydrate 3- [bis (2-chloroethyl) carbamate] 17- (dihydrogen phosphate) estradiol)</td><td>Emcyt</td><td>Pharmacia &amp; Upjohn Company</td>

<td>Etoposide phosphate (4 '- (dihydrogen phosphate) 4'-demethylepipodophyllotoxin-9- [4,6-O (R) -ethylidene- (beta) -D-glucopyranoside])</td><td>ETOPOPHOS</td><td>Bristol-Myers Squibb</td>

<td>etoposide, VP-16 (4'-demetyloepipodofilotoksyno-9- [4,6-0- (R) -ethylidene (beta) -Dglukopiranozyd])</td><td>Vepesid</td><td>Bristol-Myers Squibb</td>

<td>exemestane (6-metyloenandrosta-1,4-diene-3,17-dione)</td><td>Aromasin</td><td>Pharmacia &amp; Upjohn Company</td>

<td>Filgrastim (R-metHuG-CSF)</td><td>Neupogen</td><td>Amgen, Inc.</td>

<td>phloxuridine (intraarterial) (2'-deoxy-5-fluorouridine)</td><td>FUDR</td><td>Roche</td>

<td>fludarabine (fluorinated nucleotide analogue of the antiviral agent vidarabine, 9-bD-arabinofuranosyloadenine (ara-A))</td><td>Fludara</td><td>Berlex Laboratories, Inc., Cedar Knolls, NJ</td>

<td>Fluorouracil, 5-FU (5-fluoro-2.4 (1H, 3H) -pyrimidinedione)</td><td>Adrucil</td><td>ICN Pharmaceuticals, Inc., Humacao, Puerto Rico</td>

<td>fulvestrant (7-alpha- [9- (4,4,5,5,5-pentafluoropentylosulfinylo) nonyl] estra1,3,5- (10) -triene-3,17-beta-diol)</td><td>Faslodex</td><td>IPR Pharmaceuticals, Guayama, Puerto Rico</td>

<td>gemcitabine (2'-deoxy-2 ', 2'-difluorocytidine monohydrochloride (b-isomer))</td><td>Gemzar</td><td>Eli Lilly</td>

<td>Gemtuzumab Osogamicin (anti-CD33 hP67.6)</td><td>Mylotarg</td><td>Wyeth Ayerst</td>

<td>Goserelin acetate (acetate salt [D-Ser (But)⁶, Azgly¹⁰] LHRH; pyro-Glu-His-Trp-SerTyr-D-Ser (But) -Leu-Arg-Pro-Azgly-NH2 acetate [C59H84N18O14 (C₂H4O2) 3</td><td>Zoladex Implant</td><td>AstraZeneca Pharmaceuticals</td>

<td>hydroxyurea</td><td>Hydrea</td><td>Bristol-Myers Squibb</td>

<td>Ibritumomab Tiucsetan (immunoconjugate resulting from covalent thiourea binding between monoclonal antibody Ibritumomab and linker-chelator thiuksetan [N- [2-bis (carboxymethyl) amino] -3- (p-isothiocyanatophenyl) -propyl] [N- [2-bis (carboxymethyl) amino] 2- (methyl) ethyl] glycine)</td><td>Zevalin</td><td>Biogen IDEC, Inc., Cambridge MA</td>

EP 2 019 671 B1

<td>idarubicin ((7S-cis) -5.12-naftacenodiono-9-acetyl-7 - [(3-amino-2,3,6trideoksy- (alpha) -L-lyxo-hexopyranosyl) oxy] -7,8,9, 10tetrahydro-6,9,11-trihydroksychlorowodorek)</td><td>Idamycin</td><td>Pharmacia &amp; Upjohn Company</td>

<td>ifosfamide (3- (2-chloroethyl) -2 - [(2-chloroethyl) amino] tetrahydro2H-1,3,2-oxazaphosphite 2-oxide)</td><td>IFEX</td><td>Bristol-Myers Squibb</td>

<td>Imatinib mesylate (4 - [(4-methyl-1-piperazinyl) methyl] -N- [4-methyl-3 - [[4- (3-pyridinyl) -2-pyrimidinyl] amino] phenyl] benzamide methanesulfonate)</td><td>Gleevec</td><td>Novartis AG, Basel, Switzerland</td>

<td>Interferon alfa-2a (recombinant peptide)</td><td>Roferon-A</td><td>Hoffmann-La Roche, Inc., Nutley, NJ</td>

<td>Interferon alfa-2b (recombinant peptide)</td><td>Intron A (Lyophilized Betaseron)</td><td>Schering AG, Berlin, Germany</td>

<td>Irinotecan HCl ((4S) -4,11-diethyl-4-hydroxy-9 - [(4-piperidinopiperidine) carbonyloxy] -1H-pyran [3 ', 4': 6.7] indolizine [1,2-b] quinoline hydrochloride trihydrate 3.14 (4H, 12H) dione)</td><td>Camptosar</td><td>Pharmacia &amp; Upjohn Company</td>

<td>lenalidomide 3- (4-amino-1-oxo-1,3-dihydro-2H-isoindol-2-yl) piperidine2,6-dione</td><td>Revlimid</td><td>Celgene</td>

<td>letrozole (4,4 '- (1H-1,2,4-triazol-1-ylmethylene) dibenzonitryl)</td><td>Femara</td><td>Novartis</td>

<td>leucovorin (N- [4 - [[(2-amino-5-formyl-1,4,5,6,7,8-hexahydro-4-oxo-6-pteridinyl) methyl] amino] benzoyl] -L-glutamic acid calcium salt (1: 1))</td><td>Wellcovorin, Leucovorin</td><td>Immunex, Corp., Seattle, WA</td>

<td>Levamisol HCl ((-) - (S) -2,3,5,6-tetrahydro-6-phenylimidazo [2,1-b] thiazole monohydrochloride₁₁H₁₂N₂S ^ HCl)</td><td>Ergamisol</td><td>Janssen Research Foundation, Titusville, NJ</td>

<td>lomustine (1- (2-chloro-ethyl) -3-cyclohexyl-1-nitrosourea)</td><td>CeeNU</td><td>Bristol-Myers Squibb</td>

<td>Mecloretamine, nitrogen mustard (2-chloro-N- (2-chloroethyl) -N-methylethanamine hydrochloride)</td><td>Mustargen</td><td>Merck</td>

<td>Megestrol acetate 17 (acetyloxy) -6-methylpregna-4,6-diene-3,20-dione</td><td>Megace</td><td>Bristol-Myers Squibb</td>

<td>Melphalan, L-PAM (4- [bis (2-chloroethyl) amino] -L-phenylalanine)</td><td>Alkeran</td><td>GlaxoSmithKline</td>

<td>Mercaptopurine, 6-MP (1,7-dihydro-6H-purine-6-thione monohydrate)</td><td>Purinethol</td><td>GlaxoSmithKline</td>

<td>mesna (Sodium 2-mercaptoethanesulfonate)</td><td>Mesnex</td><td>Asta Medica</td>

<td>methotrexate (N- [4 - [[(2,4-diamino-6-pteridinyl) methyl] methylamino] benzoyl] -L-glutamic acid)</td><td>Methotrexate</td><td>Lederle Laboratories</td>

EP 2 019 671 B1

<td>methoxsalen (9-methoxy-7H-furo [3,2-g] [1] benzopyran-7-one)</td><td>Uvadex</td><td>Therakos, Inc., Way Exton, Pa</td>

<td>Mitomycin C.</td><td>Mutamycin</td><td>Bristol-Myers Squibb</td>

<td>mitomycin C.</td><td>Mitozytrex</td><td>SuperGen, Inc., Dublin, CA</td>

<td>mitotane (1,1-dichloro-2- (o-chlorophenyl) -2- (p-chlorophenyl) ethane)</td><td>Lysodren</td><td>Bristol-Myers Squibb</td>

<td>mitoxantrone (1,4-dihydroxy-5,8-bis [[2 - [(2-hydroxyethyl) amino] ethyl] amino] -9,10-anthracenedione dihydrochloride)</td><td>Novantrone</td><td>Immunex Corporation</td>

<td>Nandrolone Fenpropionate</td><td>Durabolin-50</td><td>Organon, Inc., West Orange, NJ</td>

<td>Nofetumomab</td><td>Verluma</td><td>Boehringer Ingelheim Pharma KG, Germany</td>

<td>oprelvekin (IL-11)</td><td>Neumega</td><td>Genetics Institute, Inc., Alexandria, VA</td>

<td>Oksaliloplatyna (cis - [(1R, 2R) -1,2-cyclohexanediamine-N, N '] [oxalate (2) -O, O'] platinum)</td><td>Eloxatin</td><td>Sanofi Synthelabo, Inc., NY, NY</td>

<td>paclitaxel (4,10-diacetate 2-benzoate 13-ester 5β, 20-epoxy1,2a, 4,7e, 10e, 13a-hexahydroxytax-11-en-9-one with (2R, 3S) N-benzoyl-3-phenylisoserine )</td><td>TAXOL</td><td>Bristol-Myers Squibb</td>

<td>pamidronate (bis- (3-amino-1-hydroxypropylidene) phosphonic acid disodium pentahydrate (APD))</td><td>Aredia</td><td>Novartis</td>

<td>Pegademaza (deaminase (succinimidyl monomethoxypolyethylene glycol) 11-17 -adenosine)</td><td>Adagen (Pegademase Bovine)</td><td>Enzon Pharmaceuticals, Inc., Bridgewater, NJ</td>

<td>pegaspargase (monomethoxypolyethylene glycol-succinimidyl Lasparaginase)</td><td>Oncaspar</td><td>Enzon</td>

<td>pegfilgrastim (covalent conjugate of recombinant methionyl human G-CSF (Filgrastim) and monomethoxypolyethylene glycol)</td><td>Neulasta</td><td>Amgen, Inc.</td>

<td>pentostatin</td><td>Nipent</td><td>Parke-Davis Pharmaceutical Co., Rockville, MD</td>

<td>pipobroman</td><td>Vercyte</td><td>Abbott Laboratories, Abbott Park, IL</td>

<td>Amycin, Mitramycin (antibiotic produced by Streptomyces plicatus)</td><td>Mithracin</td><td>Pfizer, Inc., NY, NY</td>

<td>Sodium Porphimer</td><td>Photofrin</td><td>QLT Photoleczniczes, Inc., Vancouver, Canada</td>

EP 2 019 671 B1

<td>procarbazine (N-isopropyl-p- (2-methylhydrazino) -ptoluamide monohydrochloride)</td><td>Matula</td><td>Sigma Tau Pharmaceuticals, Inc., Gaithersburg, MD</td>

<td>quinacrine (6-chloro-9- (1-methyl-4-diethyl-amino) butylamino-2-methoxyacridine)</td><td>Atabrine</td><td>Abbott Labs</td>

<td>rasburicase (recombinant peptide)</td><td>Elitek</td><td>Sanofi-Synthelabo, Inc.,</td>

<td>rituximab (recombinant anti-CD20 antibody)</td><td>Rituxan</td><td>Genentech, Inc., South San Francisco, CA</td>

<td>sargramostim (recombinant peptide)</td><td>Prokine</td><td>Immunex Corp</td>

<td>streptozocin (streptozocin 2-deoxy-2 - [[(methylnitrosamino) carbonyl] amino] -a (ib) -D-glucopyranose and 220 mg of anhydrous citric acid)</td><td>Zanosar</td><td>Pharmacia &amp; Upjohn Company</td>

<td>Talc (Mg₈Si4O₁0 (OH) 2)</td><td>Sclerosol</td><td>Bryan, Corp., Woburn, MA</td>

<td>tamoxifen (2-hydroxy-1,2,3-propanetricarboxylate (Z) 2- [4- (1,2-diphenyl-1-butenyl) phenoxy] -N, N-dimethylethanamine (1: 1))</td><td>Nolvadex</td><td>AstraZeneca Pharmaceuticals</td>

<td>temozolomide (3,4-dihydro-3-methyl-4-oxoimidazo [5,1-d] -as-tetrazine-8karboksyamid)</td><td>Temodar</td><td>Schering</td>

<td>Teniposide, VM-26 (9- [4.6-0- (R) -2-tenylidene- (beta) -D-glucopyranoside] 4'demetylpipodophyllotoxin)</td><td>Vumon</td><td>Bristol-Myers Squibb</td>

<td>testolactone ([dgr] -lactone 13-hydroxy-3-oxo-13,17-sekoandrosta-1,4-diene-17-acid)</td><td>Teslac</td><td>Bristol-Myers Squibb</td>

<td>Thioguanine, 6-TG (2-amino-1,7-dihydro-6H-purin-6-thione)</td><td>thioguanine</td><td>GlaxoSmithKline</td>

<td>thiotepa (1,1 ', 1'-phosphinothioilidinotrisaziridine, or tris (1aziridinyl) phosphinosulfide)</td><td>Thioplex</td><td>Immunex Corporation</td>

<td>Topotecan HCl ((S) -10 - [(dimethylamino) methyl] -4-ethyl-4,4-dihydroxy-1H-pyran [3 ', 4': 6.7] indolizino [1,2-b] quinoline 3,14- ( 4H, 12H) -dione)</td><td>Hycamtin</td><td>GlaxoSmithKline</td>

<td>toremifene (2- (p - [(Z) -4-chloro-1,2-diphenyl-1-butenyl] phenoxy) N, N-dimethylethylamine citrate (1: 1))</td><td>Fareston</td><td>Roberts Pharmaceutical Corp., Eatontown, NJ</td>

<td>Tosytumomab, I 131 Tosytumomab (recombinant mouse immunotherapeutic lambda anti-CD20 IgG2a monoclonal antibody (I 131 is a radioimmunotherapeutic antibody))</td><td>Bexxar</td><td>Corixa Corp., Seattle, WA</td>

EP 2 019 671 B1

<td>trastuzumab (recombinant anti-HER2 monoclonal IgG1 kappa antibody)</td><td>Herceptin</td><td>Grenentech, Inc.</td>

<td>Tretinoin, ATRA (completely trans retinoic acid)</td><td>Vesanoid</td><td>Roche</td>

<td>Uracil mustard</td><td>He lost Mustard Capsules</td><td>Roberts Labs</td>

<td>Valrubicin, N-trifluoroacetyl adriamycin-14-valerate ((2S-cis) -2- [1,2,3,4,6,11-hexahydro-2,5,12-trihydroxy-7-methoxy-6,11-dioxo- [ [4 2,3,6-trideoxy-3 - [(trifluoroacetyl) amino-oL-lixo-hexopyranosyl] oxyl] -2-naphthacenyl] -2-oxoethyl pentanoate)</td><td>Valstar</td><td>Anthra -&gt; Medeva</td>

<td>Vinblastine, Leurocristine (C46H56N4OK / H2SO4)</td><td>Velban</td><td>Eli Lilly</td>

<td>vincristine (C46H56N4O10 ^ H₂SO4)</td><td>Oncovin</td><td>Eli Lilly</td>

<td>vinorelbine ([R- (R *, R *) - 2,3-dihydroxybutanedione 3 ', 4'-didehydro-4'deoxy-C'-norinkaleucoblastin (1: 2) (salt)])</td><td>Navelbine</td><td>GlaxoSmithKline</td>

<td>Zoledronate, Zoledronic Acid ((1-hydroxy-2-imidazol-1-ylphosphonoethyl) phosphonic acid monohydrate)</td><td>Zometa</td><td>Novartis</td>

[0109] Anti-cancer agents also include compounds that have been identified as having anti-cancer activity but are not currently approved by the US Food and Drug Administration or other relevant agencies or are undergoing evaluation for new uses. Examples include, but are not limited to, 3-AP, 135 12-O-tetradecanoylforbol acetate, 17AAG, 852A, ABI-007, ABR-217620, ABT-751, ADI-PEG 20, AE-941, AG-013736, AGRO100, alanosine, AMG 706, G250 antibody, antineoplastones, AP23573, apazinone, APC8015, atyprymod, ATN-161, atrasenten, azacytidine, BB-10901, BCX-1777, bevacizumab, BG00001, bicalutamide, BMS 24750, , buserelin, calcitriol, CCI-779, CDB-2914, cefixime, cetuximab, CG0070, cilengitide, clofarabine, combretastatin A4 phosphate, CP-675,206, CP-724.714, CpG

7909, curcumin, decitabine, DENSPM, doxerxalciferol, E7070, E7389, ecteinascidin 743, efaproxiral, eflomitin, EKB-569, enzastaurine, erlotinib, exsulindin, fenretinid, fludarabine, fludarabine, fludapine , glufosfamide, GTI-2040, histrelin, HKI-272, homoharringtonin, HSPPC-96, hu14.18-interleukin-2, fusion protein, HuMax-CD4, iloprost, imichimod, infliximab, interleukin-12, IPI-504, irofulven, ixabepilone, lapatinib, lestaurtinib, leuprolide, immunotoxin

LMB-9, lonafarnib, luniliximab, mafosphamide, MB07133, MDX-010, MLN2704, monoclonal antibody

3F8, monoclonal antibody J591, motexafine, MS-275, MVA-MLTC1-IL2, nilutamide, nitrocamptothecin, nolatrexed dihydrochloride, nolvadex, NS-9, O6-benzylguuanine, oblimersen sodium, ONYX-015, oregovab, panitum-7 paraplatin, PD-0325901, pemetrexed, PHY906, pioglitazone, pirfenidone, pixantrone, PS-341, PSC 833, PXD101, pyrazoloacridine, R115777, RAD001, ranpimase, reec beccamycin analog, rhuAngi-roscatincine 2 , S-8184, satraplatin, SB-, 15992, SGN-0010, SGN-40, sorafenib, SR31747A, ST1571, SU011248, hydroxamic acid suberoilanilide, suramin, thalabostat, talampanel, tarichidar, temsirolimus, TGFa-PE38 immunotoxin, thalidomide, thymamazine, tefasparin trimetrexate, TroVax, UCN-1,

Valproic acid, vinflunine, VNP40101M, wolocoximab, worinostat, VX-680, ZD1839, ZD6474, zileuton and zosuchidar trihydrochloride.

[0110] For a more detailed description of anti-cancer agents and other therapeutic agents, specialists in this field are referred to many educational textbooks, including but not limited to Physician's Desk Reference and Goodman and Gilman, "Pharmaceutical Basis of Therapeutics", tenth edition , edited by Hardman et al., 2002.

[0111] The present description provides methods for administering a compound of formula II with radiation therapy. There is no limit to the types, amounts or delivery and administration systems used to deliver a therapeutic dose of radiation to an animal. For example, the animal may receive photon radiation therapy, particle beam radiation therapy, other types of radiation therapy, and combinations thereof. Radiation can be delivered to the animal using a linear accelerator or radiation is delivered using a gamma knife.

[0112] The radiation source may be external or internal to the animal. External radiation therapy is the most common and involves directing a beam of high-energy radiation to the tumor sites through the skin using, for example, a linear accelerator. Although the radiation beam is located at the tumor site, it is almost impossible to avoid exposure to normal, healthy tissue. However, external radiation is usually well tolerated by animals. Internal radiation therapy includes implantation of a radiation emitting source, such as beads, wires, granules, capsules, particles and the like, within the body at or near the tumor site, including using delivery systems that are specifically targeted for cancer cells (e.g. , using particles associated with ligands that bind to cancer cells). Such implants can be removed after therapy or left inactive in the body. Types of internal radiation therapy include, but are not limited to, brachytherapy, interstitial irradiation, cavity irradiation, radioimmunotherapy, and the like.

[0113] The animal may optionally receive radiation sensitizers (e.g., metronidazole, misonidazole, intraarterial Budr, intravenous iododeoxyuridine (IudR), nitroimidazole, 5-substituted-4-nitroimidazole, 2-Hisoindolodion, [[(2-bromoethyl]] -amino) -nitro-1H-imidazole-1-ethanol, nitroaniline derivatives, selective for hypoxia of cytotoxin with DNA affinity, halogenated DNA ligand, 1,2,4-benzotriazine oxides, 2-nitroimidazole derivatives, containing fluorine nitroazole derivatives, benzamide, nicotinamide, acridine intercalator, 5-thiotetrazole derivative, 3-nitro-1,2,4-triazole, 4,5-dinitroimidazole derivative, hydroxylated texaphrine, cisplatin, mitomycin, tiripazamine, nitrosourea, mercaptopurine, methotrexate, bleach carboplatin, epirubicin, doxorubicin, cyclophosphamide, vindesine, etoposide, paclitaxel, heating (hyperthermia), and the like), radioprotectants (e.g., cysteamine, aminoalkyl dihydrogen phosphorothioates, amifostine (WR 2721), IL-1, IL-6, and the like). Radiation sensitizers support the killing of cancer cells. Radio protective agents protect healthy tissue from the harmful effects of radiation.

[0114] Any type of radiation can be administered to an animal, as long as the radiation dose is tolerated by the patient without unacceptable negative side effects. Suitable types of radiation therapy include, for example, ionizing (electromagnetic) radiation therapy (e.g., X-rays or gamma rays) or particle beam radiation therapy (e.g., high-energy linear radiation). Ionizing radiation is defined as radiation containing particles or photons that have sufficient energy to achieve ionization, i.e., gain or loss of electrons (as described, for example, in US 5,770,581). The effects of radiation can be at least partially controlled by clinics40

EP 2 019 671 B1 The radiation dose is preferably divided into fractions for maximum effect on the target cell and reduction of toxicity.

[0115] The total dose of radiation administered to the animal is preferably about 0.01 gray (Gy) to about 100 Gy. More preferably, about 10 Gy to about 65 Gy (e.g., about 15 Gy, 20 Gy, 25 Gy, 30 Gy, 35 Gy, 40 Gy, 45 Gy, 50 Gy, 55 Gy or 60 Gy) is administered during therapy. Although the full radiation dose can be given over one day, the total dose is best divided into fractions and given over several days. Preferably, radiation therapy is administered over at least about 3 days, e.g., at least 5, 7, 10, 14, 17, 21, 25, 28, 32, 35, 38, 42, 46, 52 or 56 days (about 1-8 weeks). Accordingly, the daily dose of radiation will include approximately 1-5 Gy (e.g., approximately 1 Gy, 1.5 Gy, 1.8 Gy, 2 Gy, 2.5 Gy, 2.8 Gy, 3 Gy, 3.2 Gy, 3.5 Gy, 3.8 Gy, 4 Gy, 4.2 Gy, or 4.5 Gy), preferably 1-2 Gy (e.g., 1.5-2 Gy). The daily dose of radiation should be sufficient to induce destruction of the target cells. After being distributed in time, radiation is preferably not administered every day, thereby allowing the animal to rest and realize the effects of therapy. For example, radiation is preferably administered during 5 consecutive days, and not administered for 2 days, in each week of therapy, thereby leaving 2 days of rest per week. However, radiation may be administered before 1 day / week, 2 days / week, 3 days / week, 4 days / week, 5 days / week, 6 days / week, or the entire 7 days / week, depending on the animal's response and any potential side effects. Radiation therapy can be started at any time during the treatment period. Preferably, radiation is initiated at week 1 or week 2, and is administered for the remainder of the treatment period. For example, radiation is administered in weeks 16 or weeks 2-6 of a treatment period comprising 6 weeks of treatment, for example, a solid tumor. Alternatively, radiation is administered in weeks 1-5 or weeks 2-5 of a treatment period of 5 weeks. These sample radiotherapy regimens are just examples.

[0116] Antimicrobial therapeutic agents can also be used as therapeutic agents. Any agent that can kill, inhibit or otherwise impair the function of microbial organisms may be used, as well as any agent considered to possess such activity. Antimicrobials include, but are not limited to, natural and synthetic antibiotics, antibodies, inhibitory proteins (e.g., defensins), antisense nucleic acids, membrane disrupting agents and the like, used alone or in combination. Indeed, any type of antibiotic may be used, including, but not limited to, antibacterial agents, antiviral agents, antifungal agents, and the like.

[0117] A compound of formula II and one or more therapeutic agents or anti-cancer agents may be administered to an animal under one or more of the following conditions: at different repeats, at different durations, at different concentrations, different routes of administration, etc. the compound may be administered before the therapeutic or anti-cancer agent, e.g., at 0.5, 1.2 3.4, 5, 10, 12, or 18 hours, 1, 2, 3, 4, 5, or 6 days, 1 , 2, 3, or 4 weeks before administration of the therapeutic or anti-cancer agent. The compound may be administered after the therapeutic or anti-cancer agent, e.g., 0.5, 1, 2 3, 4, 5, 10, 12, or 18 hours, 1, 2, 3, 4, 5, or 6 days, 1, 2.3, or 4 weeks after administration of the anti-cancer agent. The compound and the therapeutic or anti-cancer agent may be administered simultaneously, but with different regimens, e.g., the compound is administered once a day and the therapeutic or anti-cancer agent is administered once a week, every two weeks, every three weeks, or every four weeks. The compound can be administered once a week, and the therapeutic or anti-cancer agent is administered once a day, once a week, every two weeks, every three weeks, or every four weeks.

[0118] Compositions include all compositions in which the compounds of the present description are included

EP 2 019 671 B1 in an amount that effectively achieves the intended purpose. Although the individual needs of the individual vary, determining the optimal ranges for effective amounts for each ingredient is within the skill of the art. Typically, the compounds may be administered to mammals, e.g., humans, orally at a daily dose of 0.0025 to 50 mg / kg body weight, or an equivalent amount of a pharmaceutically acceptable salt thereof, of a mammal being treated for disorders responsive to the induction of apoptosis. Preferably, about 0.01 to about 25 mg / kg is orally administered to treat, alleviate or prevent such disorders. For intramuscular injections, the dose is generally about half the oral dose. For example, a suitable intramuscular dose will be about 0.0025 to about 25 mg / kg, and most preferably, from about 0.01 to about 5 mg / kg.

[0119] A unit oral dose may comprise from about 0.01 to about 1000 mg, preferably about 0.1 to about 100 mg of the compound. The unit dose may be administered one or more times a day as one or more tablets or capsules, each containing from about 0.1 to about 10, suitably about 0.25 to 50 mg of the compound or its solvates.

[0120] In a topical formulation, the compound may be present at a concentration from about 0.01 to 100 mg per gram of carrier. The compound may be present at a concentration of about 0.07-1.0 mg / mL, more preferably, about 0.1-0.5 mg / mL, most preferably, about 0.4 mg / mL.

[0121] In addition to administering the compound as a raw chemical, the compounds may be administered as part of a pharmaceutical formulation containing suitable pharmaceutically acceptable carriers including excipients and excipients that facilitate processing of the compounds into preparations that can be used pharmaceutically. Preferably preparations, especially those preparations which can be administered orally or topically and which can be used for the preferred type of administration, such as tablets, dragees, lozenges and slow-release capsules, mouthwashes and gels, gels, liquid suspensions, rinses for hair, hair gels, shampoos, as well as preparations that can be administered rectally, such as suppositories, as well as appropriate solutions for administration by intravenous infusion, injection, topically or orally, contain from about 0.01 to 99 percent, preferably from about 0.25 to 75 percent of the active compound (s), together with the excipient.

[0122] The pharmaceutical compositions may be administered to any animal that may experience beneficial effects of the compounds of the invention. The main among such animals are mammals, e.g., humans, although the invention is not intended to be limited in this way. Other animals include veterinary animals (cows, sheep, pigs, horses, dogs, cats and the like).

[0123] The compounds and their pharmaceutical compositions can be administered by any means that achieve their intended purpose. For example, administration may be by parenteral, subcutaneous, intravenous, intramuscular, intraperitoneal, transdermal, buccal, intrathecal, intracranial, intranasal or local routes. Alternatively or simultaneously, administration may occur by oral route. The dosage used will depend on the age, health and weight of the recipient, the type of concomitant therapy, if any, the frequency of therapy and the nature of the effect desired.

[0124] Pharmaceutical preparations are prepared in a manner that is known per se, for example, by means of conventional mixing, granulating, dragee-forming, dissolving or lyophilizing. Thus, pharmaceutical preparations for oral use can be obtained by combining the active compounds with solid excipients, optionally grinding the resulting mixture and processing the mixture of granules, after adding suitable auxiliaries, if desired or necessary, to obtain tablets or dragee cores.

[0125] Suitable excipients are, in particular, fillers such as sugars, for example lactose or

Sucrose, mannitol or sorbitol, cellulose preparations and / or calcium phosphates, for example tricalcium phosphate or calcium hydrogen phosphate, as well as binding agents such as starch paste, using, for example, corn starch, wheat starch, rice starch, starch potato, gelatin, tragacanth, methyl cellulose, hydroxypropyl methyl cellulose, sodium carboxymethyl cellulose, and / or polyvinylpyrrolidone. If desired, disintegrants such as the above-mentioned starches and also carboxymethyl starch, cross-linked polyvinylpyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate may be added. Excipients are, above all, fluidity regulating agents and lubricants, for example silica, talc, stearic acid or its salts, such as magnesium stearate or calcium stearate, and / or polyethylene glycol. Dragee cores are provided with suitable coatings, which, if desired, are resistant to gastric juices. For this purpose, concentrated sugar solutions which may optionally contain gum arabic, talc, polyvinylpyrrolidone, polyethylene glycol and / or titanium dioxide, lacquer solutions and suitable organic solvents or solvent mixtures can be used. To produce coatings resistant to gastric juices, solutions of suitable cellulose preparations such as cellulose acetate phthalate or hydroxypropyl methyl cellulose phthalate are used. Dyes or pigments may be added to the tablets or dragee coatings, for example for identification or to determine the combination of active compound doses.

[0126] Other pharmaceutical preparations that can be used orally include sealable capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer such as glycerol or sorbitol. The sealable capsules may contain active compounds in the form of granules, which may be mixed with fillers such as lactose, binders such as starches, and / or lubricants such as talc or magnesium stearate, and optionally stabilizing agents. In soft capsules, the active compounds are preferably dissolved or suspended in suitable liquids, such as liquid fat or liquid paraffin. In addition, stabilizing agents may be added.

[0127] Possible pharmaceutical preparations that can be used rectally include, for example, suppositories, which consist of a combination of one or more active compounds with a suppository base. Suitable suppository bases are, for example, natural or synthetic triglycerides or paraffin hydrocarbons. In addition, it is also possible to use gelatin rectal capsules, which consist of a combination of active compounds with a base. Possible base materials include, for example, liquid triglycerides, polyethylene glycols, or paraffin hydrocarbons.

[0128] Suitable formulations for parenteral administration include aqueous solutions of the active compounds in water-soluble form, for example, water-soluble salts and basic solutions. In addition, suspensions of the active compounds may be administered as appropriate oily injection suspensions. Suitable lipophilic solvents or excipients include liquid fats, for example, sesame oil, or synthetic fatty acid esters, for example, ethyl oleate or triglycerides or polyethylene glycol 400. Aqueous injection suspensions may contain substances that increase the viscosity of the suspension, including, for example, sodium carboxymethyl cellulose, sorbitol, and / or dextran. Optionally, the suspension may also contain stabilizing agents.

[0129] Topical compositions are preferably formulated as oils, creams, lotions, ointments and the like by choosing appropriate carriers. Suitable carriers include vegetable or mineral oils, white petrolatum (white soft paraffin), branched chain fats or oils, animal fats and high molecular weight alcohol (greater than C12). Preferred carriers are those in which the active ingredient is soluble. Emulsifiers, stabilizing agents, moisturizing agents and antioxidants may also be present, as well as agents that provide color or fragrance, if desired. In addition, in these places43

Skin penetration enhancers can be used in the formulations. Examples of such elevating agents can be found in US Patent Nos. 3,988,816 and 4,444,762.

[0130] Creams are preferably formulated from a mixture of mineral oil, self-emulsifying beeswax and water in which the active ingredient is dissolved in a small amount of oil such as almond oil. A typical example of such a cream is a cream comprising about 40 parts water, about 20 parts beeswax, about 40 parts mineral oil and about 1 part almond oil. [0131] Ointments can be formulated by mixing a solution of the active ingredient in vegetable oil such as almond oil with warm soft paraffin and allowing the mixture to cool. A typical example of such an ointment is an ointment that contains about 30% by weight of almond oil and about 70% of white soft paraffin. [0132] Lotions may conveniently be prepared by dissolving the active ingredient in a suitable high molecular weight alcohol such as propylene glycol or polyethylene glycol.

EXAMPLE 1

Synthesis of Smac divalent mimetics [0133] General methods: NMR spectra were recorded at a proton frequency of 300 MHz. Chemical shifts¹H is given relative to Me4Si (0.00 ppm), CHCl3 (7.26 ppm), CD2HOD (3.31 ppm), or DHO (4.79 13 ppm) as internal standards. Chemical shifts¹³C is reported relative to CDCl3 (77.00 ppm), CD3OD (49.00 ppm), or 1,4-dioxane (67.16 ppm) as internal standards. Optical twists were measured at room temperature.

General procedure A (condensation):

[0134] To a solution of two substrates in CH2Cl2 (20 mg / ml for minority substrate) was added EDC (1.1 equivalents per amino group), HOBt (1.1 equivalents per amino group) and N, N-diisopropylethylamine (4 equivalents per amino group) at 0 ° C with stirring. The mixture was stirred at room temperature for eight hours and then concentrated. The residue was purified by chromatography to give the product.

General procedure B (click chemistry):

[0135] To the CuSO4 solution (10 mg / ml), sodium (+) - L-ascorbate (2 equivalents) was added. The mixture was shaken until it turned bright yellow. To the solution of two substrates in acetonitrile or 2-methylpropanol (20 mg / ml for the minority substrate) was added a previously prepared mixture of CuSO4-L-ascorbate sodium (0.1 equivalent of CuSO4 per equivalent of minority substrate). The mixture was stirred at room temperature overnight and then extracted with dichloromethane three times. The combined organic layer was washed with brine, dried over Na2SO4 and concentrated. The residue was purified by chromatography to give the product.

General procedure C (Boc deprotection):

[0136] To a solution of the substrate in methanol (20 mg / ml) was added a solution of HCl in 1,4-dioxane (4 M, 10-20 equivalents per Boc). The solution was stirred at room temperature overnight and then concentrated to give the product.

General procedure C (Boc deprotection):

[0137] To a solution of the substrate in methanol (20 mg / mL) was added a solution of HCl in 1,4-dioxane (4 M, 10-20 equivalents per Boc). The solution was stirred at room temperature overnight and then concentrated to give the product.

EXAMPLE 2

Synthesis of DQ-24, SH-143, SH-155 and SH-142 [0138] Compounds DQ-24, SH-143, SH-155 and SH-142 were synthesized according to Scheme I.

EP 2 019 671 B1

Diagram I:

<img file="PL2019671T3_D0025.tif" />

Reagents and conditions: (a) i. Trifluoroacetic anhydride, Et3N, CH3Cl2, room temperature; ii. NaHCO3, MeOH, 95%; (b) NaH, propargyl bromide, DMF. 92%; (c) i. MsCl, Et3N; ii. NaN3, DMF, 100 ° C, 85% in two stages; (d) i. 3. CuSO4. (+) - sodium L-ascorbate, CH3CN-H2O 3: 1; ii. 2N LiOH, 1,4-dioxane-H2O 1: 1,

74% in two stages; (e) i. NaH, benzyl-2-bromoethyl ether: ii. 10% Pd-C, H2, MeOH; iii. MsCl, Et3N; iv. NaN3, DMF, 65% in four stages; (f) i. 3. CuSO4, (+) - sodium L-ascorbate, CH3CN-H2O 3: 1; ii. 2N LiOH, 1,4-dioxane-H2O 1: 1, 72% in two steps; (g) propargyl ether (5 equivalents), CuSO4, (+) - sodium L-ascorbate, CH3CN-H2O 3: 1, 69%; (h) i. 8. CuSO4, (+) - sodium L-ascorbate,

CH3CN-H2O 3: 1, ii. 4N HCl in 1,4-dioxane, MeOH, 95%; (i) NaH, propargyl bromide, DMF, 82%; (J)

i. 8 (2.2 equivalents). CuSO4, (+) - Sodium L-ascorbate, CH3CN-H2O 3: 1. ii. 4N HCl in 1,4-dioxane, MeOH, 62% in two steps.

[0139] Selective protection of the amino group in L-phenylglycinol 1 with trifluoroacetic anhydride gave alcohol 2. Alkylation with 2 propargyl bromide gave alkyne 3. Reaction 2 with methanesulfonyl chloride followed by substitution of the resulting NaN3 mesylate gave azide 4. Cycloaddition 3 and 4 under catalysis CuSO4 - (+) - L-ascorbate sodium, followed by removal of the trifluoroacetyl groups gave diamine 5.

[0140] Alkylation with 2-benzyl-2-bromoethyl ether followed by hydrolysis of the benzyl protecting group gave alcohol. The reaction of this alcohol with methanesulfonyl chloride followed by substitution of the resulting NaN3 mesylate gave azide 6. Cycloaddition of 6 out of 3 under catalysis with CuSO4 - (+) - sodium lascorbate, followed by removal of the trifluoroacetyl groups gave diamine 7.

[0141] Compound 8 was synthesized according to our previously mentioned method (Sun et al., Tetrahedron Letters, 46: 7015 (2005)). Cycloaddition of compound 8 with an excess of propargyl ether (5-10 equivalents) gave alkyne 9. Cycloaddition of 9 and 8 under catalysis with CuSO4 - (+) - L-ascorbate sodium, followed by removal of the Boc protecting groups gave diamine 10.

[0142] Compound 11 was synthesized according to our previously mentioned method (Sun et al., Tetrahedron Letters, 46: 7015 (2005)). Alkylation with 11 propargyl bromide gave alkyne 12. Cycloaddition 2.2 equals 45

In this case, 8 with 1 equivalent of 12 under CuSO4 - (+) - L-ascorbate catalysis conditions, followed by removal of the Boc protecting groups gave diamine 13.

Diagram II:

<img file="PL2019671T3_D0026.tif" />

Reagents and conditions: (a) 10% Pd-C, MeOH, H2, 100%; (b) i. diamine, EDC, HOBt, N, N-diisopropylethylamine, CH 2 Cl 2; ii. 4N HCl in 1,4-dioxane, MeOH; iii. LN-Boc-N-methylalanine, EDC, HOBt, N, N-diisopropylethylamine, CH2Cl2; iv. 4N HCl in 1,4-dioxane.

[0143] Compound 14 can be synthesized according to the methods given in the literature (Duggan et al., Org. Biomol. Chem., 3: 2287 (2005)) (Scheme II). Reduction of the CC double bond and hydrolysis of the benzyl ester in compound 14 gave acid 15. Condensation of 2.2 equivalents with the diamines above, respectively, followed by removal of the Boc protecting groups gave four ammonium salts. Condensation of these salts with LN-Boc-N-methylalanine, respectively, followed by deprotection of the Boc protecting groups gave bivalent Smac DQ-24, SH-143, SH-142 and SH-155 mimetics. The yield of the synthesis scheme for each compound is shown in Table 2.

TABLE 2

<td>Name</td><td>Efficiency (%, in four stages)</td>

<td>DQ-24</td><td> 61</td>

<td>SH-143</td><td> 62</td>

<td>SH-155</td><td> 59</td>

<td>SH-142</td><td> 55</td>

<img file="PL2019671T3_D0027.tif" />

[0144] ¹H NMR (300 MHz, CDCl3): δ 7.50 (s, 1H), 7.38-7.20 (m, 3H), 7.28-7.20 (m, 2H), 7.26-7 , 05 (m, 5H), 4.98-4.75 (m, 2H), 4.23 (s, 2H), 4.36 (m, 2H), 3.50 (m, 2H); ¹³C NMR (75 MHz, CDCl3): δ 143.78, 133.54, 132.53, 130.26, 129.97, 129.70, 129.62, 127.47, 127.10, 126.40, 69.89, 63.23, 54.67, 54.60, 52.40.

<img file="PL2019671T3_D0028.tif" />

[0145] ¹H NMR (300 MHz, CDCl3): δ 7.59 (s, 1H), 7.32-7.22 (m, 3H), 7.22-7.13 (m, 5H), 7.12-7 , 05 (m, 2H), 4.55-4.29 (m, 6H), 3.85-3.75 (m, 1H), 3.75-3.56 (m, 5H); ¹³C NMR (75 MHz, CDCl3): 8 143.51, 133.88, 133.63, 129.83, 129.61, 129.52, 129.38, 127.36, 127.32, 125.59, 70.89, 70.20, 69.25, 63.42, 54.62, 54.39,

EP 2 019 671 B1

50,49.

<img file="PL2019671T3_D0029.tif" />

[0146] ¹H NMR (300 HMz, CDCl3): δ 7.55 (s, 1H), 7.40-7.08 (m, 9H), 5.90 (brs, 1H), 5.24 (brs, 1H), 4.74 (s, 2H), 4.34 (t, J = 7.1 Hz, 2H), 4.23 (d, J = 2.4 Hz, 1H), 2.96 (t, J = 7 , 3 Hz, 2H), 2.49 (t, J = 2.4 Hz, 1H), 2.22 (m, 2H), 1.43 (brs, 9H); ¹³C NMR (75 HMz, CDCl3): δ 154.93, 144.26, 141.98, 140.15, 139.04, 128.54,

128,50, 127,36, 127,22, 127,07, 122,56, 79,67, 79,16, 74,89, 62,91, 58,06, 57,37, 49,37, 31,94, 31,44, 28,25.

<img file="PL2019671T3_D0030.tif" />

[0147] ¹H NMR (300 MHz, CDCl3): δ 7.52 (s, 2H), 7.38-7.12 (m, 10H), 7.08-6.99 (m, 4H), 6.92-6 , 79 (m, 4H), 10 5.42 (s, 2H), 4.40 (s, 4H), 4.02 (m, 4H), 2.18 (m, 4H), 1.78 (m , 4H); ¹³C NMR (75 MHz, CDCl3): δ 144.10,

139,59, 137,31, 132,04, 131,80, 131,72, 129,70, 129,64, 127,47, 65,29, 60,60, 52,51, 34,08, 33,18.

<img file="PL2019671T3_D0031.tif" />

[0148] ¹H NMR (300 MHz, CDCl3): δ 7.40-7.18 (m, 5H), 7.15 (brs, 4H), 6.45 (brs, 1H), 4.24 (d, J = 2 , 4 Hz, 2H), 3.90 (d, J = 1.9 Hz, 2H), 3.55 (t, J = 6.3 Hz, 2H), 2.72 (dd, J = 8.9 , 7.4 Hz, 2H), 2.77 (t, J = 2.4 Hz, 1H),

2.04 (t, J = 1.9 Hz, 1H), 1.92 (m, 2H), 1.43 (brs, 9H); ¹³C NMR (75 MHz, CDCl3): δ 155.17, 140.92, 139.85,

136,99, 128,76, 128,53, 128,40, 128,22, 127,22, 80,68, 80,58, 79,86, 74,19, 70,26, 69,16, 58,01,34,75,31,79,30,99,28,23.

<img file="PL2019671T3_D0032.tif" />

[0149] ¹H NMR (300 MHz, D2O): δ 7.50 (s, 1H), 7.33 (s, 1H), 7.23-7.03 (m, 17H), 6.99-6.92 (m , 6H), 6.8020 6.68 (m, 4H), 5.38 (s, 1H), 5.34 (s, 1H), 5.10 (s, 1H), 4.08 (s, 2H ), 4.08 (t, J = 6.8 Hz, 1H), 3.99 (s, 2H), 3.92 (t, J = 6.8 Hz, 2H), 3.02 (t, J = 6.2 Hz, 2H), 2.30 (t, J = 7.3 Hz, 2H), 2.17 (t, J = 7.3 Hz, 2H), 2.11 (t, J = 7 , 3 Hz, 2H), 1.89-1.80 (m, 2H), 1.78-1.62 (m, 2H), 1.48-1.35 (m, 2H); ¹³C NMR (75 MHz, D2O): δ 163.46,

162,99, 144,04, 143,16, 141,81, 141,56, 137,61, 137,05, 134,70, 129,75, 129,64, 129,57, 129,54, 129,41, 129,28, 127,64, 127,45, 127,28, 127,16, 126,49, 125,00, 118,54, 114,67, 68,74, 65,33, 62,55, 58,06, 50,20,

49,98, 40,52, 31,70, 31,60, 30,98, 30,67, 30,42, 29,96.

<img file="PL2019671T3_D0033.tif" />

EP 2 019 671 B1 [0150] ¹H NMR (300 MHz, D2O): δ 7.77 (s, 1H), 7.78-7.21 (m, 10H), 5.32 (m, 1H), 4.91 (m, 1H), 4.75-4.56 (m, 4H), 4.50 (s, 2H), 4.33 (m, 1H), 4.17 (m, 3H), 3.79 (m, 2H), 3 , 60 (m, 2H), 2.55 (s, 3H), 2.53 (s, 3H), 2.257.80 (m, 4H), 1.79-1.61 (m, 7H), 1 , 60-1.45 (m, 9H), 1.40-1.38 (m, 4H), 1.37 (d, J = 7.5 Hz, 3H), 1.33 (d, J = 7 , 2 Hz, 3H); ¹³C NMR (75 MHz, D2O): δ 173.82, 173.65, 172.34, 172.29, 144.94, 138.64, 137.46, 129.41, 129.25, 128.87, 128.23, 126.91, 126.88, 125.73, 72.14, 63.14, 62.15, 61.16, 60.96, 57.28, 54.14, 53.64, 53, 34, 51.10, 35.94, 32.94, 32.33, 31.31, 27.95, 25.86.291.991.60.

<img file="PL2019671T3_D0034.tif" />

[0151] ¹H NMR (300 MHz, D2O): δ 7.50 (s, 1H), 7.30-7.08 (m, 10H), 4.95-4.80 (m, 3H), 4.75 (m , 1H), 4.47 (s, 2H), 4.39 (m, 2H), 4.32-4.10 (m, 4H), 4.35-4.08 (m, 4H), 3, 66-3.50 (m, 4H), 2.55 (s, 6H), 2.22-1.43 (m, 24H), 1.39 (m, 6H); ¹³C NMR (75 MHz, D2O): δ 173.71, 172.30, 169.52, 143.92, 138.73, 129.07, 128.18, 126.96, 125.53, 72.87, 72.32, 68.99, 63.36, 62.21, 62.14, 61.05, 57.20, 53.40, 53.24, 51.14, 50.42, 35.96,

32,99, 32,32, 31,32, 27,89, 25,08, 21,94, 15,63.

<img file="PL2019671T3_D0035.tif" />

[0152] ¹H NMR (300 MHz, D2O): δ 7.29 (s, 2H), 7.10-6.92 (m, 10H), 6.85 (d, J = 8.0 Hz, 4H), 6, 58 (d, J = 8.0 Hz, 4H), 5.78 (s, 2H), 4.65 (m, 2H), 4.38 (s, 4H), 4.22 (m, 2H), 4.08 (m, 2H), 3.95-3.73 (m, 6H), 2.55 (s, 6H), 2.21-1.28 (m, 36H); ¹³C NMR (75 MHz, D2O): δ 179.33, 172.64, 172.07, 144.32, 141.56, 140.08, 139.74, 129.15, 127.99, 127.69, 127.53, 124.85, 63.13, 62.04, 61.01, 57.48, 57.30, 51.12, 49.98, 36.03, 33.27, 32.47, 31, 77, 31.44, 31.22, 27.87, 25.25, 21.99, 15.78.

<img file="PL2019671T3_D0036.tif" />

[0153] ¹H NMR (300 MHz, CDCl3): δ 7.92 (brd, J = 8.4 Hz, 2H), 7.82 (brd, J = 8.4 Hz, 2H), 7.49 (s, 1H) , 7.45-7.06 (m, 28H), 6.21 (s, 1H), 6.19 (s, 1H), 4.90 (m, 2H), 4.85 (s, 1H), 4.75 (m, 2H), 4.62 (s, 2H), 4.33 (t, J = 7.1 Hz, 4H), 4.20 (m, 2H), 3.82 (s, 2H ), 3.49 (t, J = 7.1 Hz, 2H), 3.06 (m, 2H), 2.70-2.55 (m, 8H), 2.38 (s, 6H), 2 , 30-1.35 (m, 28H), 1.30 (d, J = 6.9 Hz, 6H); ¹³C NMR (75 MHz, CDCl3): δ 174.16, 172.05, 169.69, 145.37, 141.64, 140.60, 139.70, 139.15, 139.11, 128.60, 128.56, 128.50, 127.59, 127.47, 127.32, 127.29, 127.27, 127.02, 122.22, 121.52, 69.87, 66.39, 64, 35, 60.15, 59.75, 59.18, 56.78, 53.41, 49.41, 49.26, 42.65, 36.71, 35.97, 35.12, 32.03, 31.59, 31.55, 31.09, 24.91, 24.06, 23.20, 19.44.

EXAMPLE 3

Synthesis of SH-156, SH-158, SH-159, SH-164, SH-165, SH-166 and SH-167 [0154] Compounds SH-156, SH-158, SH-159, SH-164, SH- 165, SH-166 and SH-167 were synthesized according to Scheme III.

Diagram III:

EP 2 019 671 B1

<img file="PL2019671T3_D0037.tif" />

Reagents and conditions: (a) i. 17, EDC, HOBt, N, N-diisopropylethylamine, CH2Cl2; ii. 4 N HCl in 1,4-dioxane, MeOH; iii. LN-Boc-N-methylalanine, EDC, HOBt, CH2Cl2, 78% in three stages; (b) i. diazide, CuSO4, (+) - sodium L-ascorbate, t-BuOH-H2O 3: 1, ii. 4N HCl in 1,4-dioxane, MeOH.

[0155] Chiral amine 17 can be prepared according to methods given in the literature of compound 16 [Messina, et al., J. Org. Chem., 64: 3767 (1999)]. Acid condensation with chiral amine 17 followed by deprotection of the Boc HCl protecting group in methanol gave the ammonium salt. Condensation of this salt with L-NBoc-N-methylalanine gave intermediate 18. Cycloaddition of 18 with the appropriate diazide under catalysis of CuSO4 - (+) - L-ascorbate sodium, followed by deprotection of the Boc protecting group, gave specific bivalent Smac SH-156, SH-158, SH-159, SH-164, SH-165 mimetics. , SH-166 and SH-167. The yield of the synthesis scheme for each compound is shown in Table 3.

TABLE 3

<td>Name</td><td>Efficiency (%, in two stages)</td>

<td>SH-156</td><td> 63</td>

<td>SH-158</td><td> 65</td>

<td>SH-159</td><td> 61</td>

<td>SH-164</td><td> 62</td>

<td>SH-165</td><td> 59</td>

<td>SH-166</td><td> 58</td>

<td>SH-167</td><td> 59</td>

<img file="PL2019671T3_D0038.tif" />

[0156] ¹H NMR (300 MHz, CDCl3): δ 7.75 (brd, J = 8.5 Hz, 1H), 7.50-7.44 (m, 2H), 7.38-7.23 (m, 3H ), 6.90 (brs, 1H), 5.95 (dd, J = 8.5, 2.4 Hz, 1H), 4.80 (m, 1H), 4.64 (dd, J = 8, 4, 6.2 Hz, 1H), 4.60 (brm, 1H), 4.15 (m, 1H), 2.88 (s, 3H), 2.62 (m, 1H), 2.53 ( d, J = 2.4 Hz, 1H), 2.20-1.70 (m, 5H), 1.51 (brs, 9H), 1.56-1.05 (m, 9H); ¹³C NMR (75 MHz, CDCl3): δ 171.99, 170.95, 169.73, 138.96, 129.03, 128.55, 127.53, 82.13, 73.25, 60.03, 59.47, 50.32, 45.21, 36.94, 36.30, 32.36, 30.48, 28.80, 25.29, 24.42, 23.45, 14.16.

EP 2 019 671 B1

<img file="PL2019671T3_D0039.tif" />

[0157] ¹H NMR (300 MHz, D2O): δ 7.58 (s, 2H), 7.29-7.13 (m, 10H), 6.08 (s, 2H), 4.70 (m, 2H), 4.38 (m, 4H), 4.27 (m, 2H), 4.22 (m, 2H), 3.85 (m, 2H), 3.73 (m, 4H), 3.32 (m , 4H), 3.25 (m, 4H), 2.58 (s, 6H), 2.257.48 (m, 22H), 1.40 (d, J = 7.0 Hz, 6H), 1.39 (m, 2H); ¹³C NMR (75 MHz, D2O): δ 173.36, 172.32, 169.56,

148,12, 139,21, 129,32, 128,54, 127,40, 124,51, 69,98, 69,75, 68,97, 62,07, 61,07, 57,20, 51,13, 50,46,

50,41, 35,94, 33,01, 32,35, 31,31, 27,81, 25,07, 21,92, 15,63.

<img file="PL2019671T3_D0040.tif" />

[0158] ¹H NMR (300 MHz, D2O): 8 7.56 (s, 2H), 7.29-7.13 (m, 10H), 6.05 (s, 2H), 4.70 (m, 2H), 4.38-4.14 (m, 8H), 3.85 (m, 2H), 3.63 (m, 4H), 3.35 (s, 4H), 2.55 (s, 6H), 2 , 22-1.45 (m, 22H), 1.42 (d, J = 7.1 Hz, 6H),

1.40 (m, 2H); ¹³C NMR (75 MHz, D2O): δ 173.31, 172.28, 169.54, 148.15, 139.23, 129.30, 128.53, 127.38,

124,45, 69,85, 68,90, 62,05, 61,05, 57,20, 51,12, 50,39, 35,93, 33,03, 32,34, 31,32, 27,78, 25,07, 21,93, 15,63.

<img file="PL2019671T3_D0041.tif" />

[0159] ¹H NMR (300 MHz, D2O): δ 7.51 (s, 2H), 7.28-7.04 (m, 10H), 6.04 (s, 2H), 4.70 (m, 2H), 4.39-4.15 15 (m, 8H), 3.82 (m, 2H), 3.70 (m, 4H), 2.56 (s, 6H), 2.20-1.45 (m , 22H), 1.40 (d, J = 6.9 Hz, 6H), 1.38 (m, 2H);

¹³C NMR (75 MHz, D2O): δ 173.28, 172.22, 169.52, 148.14, 139.30, 129.31, 128.48, 127.33, 124.36, 68.99, 62.04, 61.06, 57.21, 51.10, 50.42, 50.35, 35.93, 33.06, 32.35, 31.32, 27.81, 25.09, 21, 93, 15.64.

<img file="PL2019671T3_D0042.tif" />

[0160] ¹H NMR (300 MHz, D2O): δ 7.40 (s, 2H), 7.15-6.85 (m, 10H), 6.65 (s, 4H), 6.08 (s, 2H), 4.65 (m, 2H), 4.32 (m, 2H), 4.08 (m, 2H), 3.92-3.74 (m, 6H), 2.84 (m, 4H), 2.54 (s, 6H), 2.28-1.04 (m, 38H); ¹³C NMR (75

MHz, D2O): δ 172.24, 171.83, 169.34, 148.50, 139.56, 139.41, 129.07, 128.53, 127.43, 122.92, 61.70, 60, 72, 57.20, 51.23, 50.86, 50.15, 36.08, 34.96, 34.64, 31.34, 29.61, 28.63, 28.57, 28.20, 27.60, 25.20, 15.71.

EP 2 019 671 B1

<img file="PL2019671T3_D0043.tif" />

[0161] ¹H NMR (300 MHz, D2O): δ 7.55 (s, 2H), 7.19-7.05 (m, 10H), 6.85 (s, 4H), 5.98 (s, 2H), 5.15 (s, 4H), 4.65 (m, 2H), 4.25 (t, J = 7.2 Hz, 2H), 4.10 (m, 2H), 3.82 (m, 2H ), 2.54 (s, 6H), 2.12-1.20 (m, 30H); ¹³C NMR (75 MHz, D2O): δ 173.04, 172.16, 169.49, 148.55, 139.08, 135.38, 129.27, 128.82, 128.51, 127.40, 123,90,

62,92, 60,92, 57,19, 53,62, 51,05, 50,39, 35,88, 33,05, 32,28, 31,32, 27,66, 25,06, 21,91, 15,64.

<img file="PL2019671T3_D0044.tif" />

[0162] ¹H NMR (300 MHz, D2O): 8 7.59 (s, 2H), 7.28-7.10 (m, 10H), 6.02 (s, 2H), 4.65 (m, 2H), 4.29 (m, 2H), 4.22-4.08 (m, 6H), 3.82 (m, 2H), 2.53 (s, 6H), 2.20-1.42 (m, 26H), 1.40 (d, J = 7.1 Hz, 6H), 1.35 (m, 2H); ¹³C NMR (75 MHz, D2O): 8 173.35, 172.29, 169.53, 148.20, 139.07, 129.31, 128.54, 127.36, 124.06, 62.05,

61,03, 57,19, 51,11, 50,32, 50,05, 35,92, 33,02, 32,33, 31,31, 27,76, 26,68, 25,06, 21,93, 15,63.

<img file="PL2019671T3_D0045.tif" />

[0163] ¹H NMR (300 MHz, D2O): δ 7.62 (s, 2H), 7.28-7.12 (m, 10H), 6.07 (s, 2H), 4.65 (m, 2H), 4.30 (t, J = 8.8 Hz, 2H), 4.25-4.10 (m, 6H), 3.84 (m, 2H), 2.55 (s, 6H), 2.23 -1.95 (m, 4H), 1.95-1.45 (m, 22H), 1.40 (d, J = 7.0 Hz, 6H), 1.35 (m, 2H), 1, 05-0.85 (m, 8H); ¹³C NMR (75 MHz, D2O): δ 173.33, 172.28, 169.55, 148.22,

139,18, 129,30, 128,54, 127,43, 123,96, 62,04, 61,02, 57,18, 51,10, 50,72, 50,44, 35,94, 33,03, 32,34,

31,32, 29,35, 27,86, 25,40, 25,09, 21,90, 15,64.

EXAMPLE 4

Synthesis of SH-153 and SH-172 [0164] Compounds SH-153 and SH-172 were synthesized according to Schemes IV and V.

Diagram IV:

EP 2 019 671 B1

<img file="PL2019671T3_D0046.tif" />

Reagents and conditions: (a) 9-BBN, THF, then H2O2 (35% in water), 3 N NaOH; (b) Dess-Martin periodinate, CH2Cl2, yield 33%, yield 62%; (c) Dess-Martin periodinate, CH2Cl2, 96%; (d) NaBH3CN, MeOH, H2SO4 (catal.), 94%.

[0165] Borohydriding of the double CC bond in compound 14 by treatment with 9-BBN followed by oxidation of the resulting borane with basic H2O2 gave a mixture of four alcohols. Alcohol 19 can be separated from the other three isomers by chromatography and its structure confirmed by X-ray analysis. Oxidation of the mixture of the remaining three isomers with Dess-Martin periodinate gave two ketones 20 and 21, which can be separated by chromatography. Reduction of the NaBH3CN ketone in the presence of a catalytic amount of H2SO4 gave alcohol 22 as a single isomer. Oxidation of alcohol 19 with Dess-Martin periodinate also gave ketone 20, so the structure of alcohol 22 was also confirmed.

EP 2 019 671 B1

Scheme V:

<img file="PL2019671T3_D0047.tif" />

Reagents and conditions: (a) i. 10% Pd-C, H2, MeOH; ii. (R) - (-) - 1,2,3,4-tetrahydro-1-naphthylamine, EDC, HOBt, N, N-diisopropylethylamine, CH 2 Cl 2; iii. MsCl, N, N-diisopropylethylamine, CH2Cl2; iv. NaN 3,

DMF; (b) propargyl ether (5 equivalents), CuSO4, (+) - sodium L-ascorbate, AcCN: t-BuOH: H2O 2: 2: 1, room temperature; (c) 23 or 24, CuSO4, (+) - sodium L-ascorbate, t-BuOH: H2O 1: 1, room temperature; (d) i. 4 N HCl in 1,4-dioxane, MeOH, ii. LN-Boc-N-methylalanine, EDC, HOBt, N, N-diisopropylethylamine, CH2Cl2; iii. 4 N HCl in 1,4-dioxane, MeOH.

[0166] Hydrolysis of benzyl esters at 19 and 22 followed by condensation of the resulting acid with (R) - (-) 1,2,3,4-tetrahydro-1-naphthylamine gave two amides (Scheme V). The reaction of these two amides with methanesulfonyl chloride followed by substitution of the resulting two NaN3 mesylates gave two azides 23 and 24. The cycloaddition of these two azides with an excess of propargyl ether under conditions of CuSO4 - (+) - sodium lascorbate gave two alkynes 25 and 26. Cycloaddition 23 and 24 with these two alkynes respectively gave compounds 27 and 28. Removal of the Boc protecting groups in these two compounds followed by condensation with LN-Boc-N-methylalanine gave two amides. Removal of the Boc protecting groups in these two amides gave SH-153 and SH-172, respectively.

<img file="PL2019671T3_D0048.tif" />

[0167] To a solution of compound 14 (1.25 g, 3 mmol) in 50 ml dry THF was added 9 ml of a 9-BBN solution (0.5 M in THF, 4.5 mmol). After heating the solution at reflux for 12 h, 1.5 ml of 3 M NaOH solution and 2 ml of H2O2 solution (35% in water) at 0 ° C. were added dropwise. After warming to room temperature and stirring for 2 h, the mixture was extracted with ethyl acetate three times. The combined organic layer was dried over Na2SO4 and then concentrated. The residue was purified by chromatography to give compound 19 (330 mg, 25%) and a mixture of the three remaining isomers (580 mg, 45%).

[0168] Chemical data for compound 19: ¹H NMR (300 MHz, CDCl3): δ 7.40-7.28 (m, 5H), 5.43 (brd, J = 7.8 Hz, 1H), 5.28, 5.18 (ABq, J = Hz, 2H), 4.67 (t, J = 8.4 Hz, 1H), 4.65 (m, 1H), 4.20 (m, 1H), 3.96 (m, 1H), 2 , 45-1.60 (m, 10H), 1.38 (brs, 9H); ¹³C NMR (75 MHz, CDCl3): δ 172.38, 170.83, 155.13, 135.39, 128.58, 128.40, 128.34, 79.70, 70.63, 67.14, 60.13, 56.16, 50.70, 45.22, 32.67, 31.70, 28.35, 27.34.

<img file="PL2019671T3_D0049.tif" />

[0169] To a solution of the mixture of the three isomers obtained above (570 mg, 1.3 mmol) in 15 mL of CH2Cl2 was added Dess-Martin periodinane (660 mmol, 1.56 mmol) at room temperature. The mixture was stirred at the same temperature for 2 h and then concentrated. The residue was purified by chromatography to give compound 20 (160 mg, 28%) and 21 (330 mg, 58%). Compound 19 can be oxidized to compound 20 in the same manner.

[0170] Chemical data for compound 20: ¹H NMR (300 MHz, CDCl3): δ 7.40-7.28 (m, 5H), 5.42 (brd, J = 8.2 Hz, 1H), 5.28, 5.18 (ABq, J = 12.2 Hz, 2H), 4.62 (t, J = 8.4 Hz, 1H), 4.37 (m, 1H), 3.13 (m, 1H), 3.02 (t, 1H ), 2.50-1.98 (m, 8H), 1.83 (m, 1H), 1.60 (m, 1H); 1.38 (brs, 9H);¹³C NMR (75 MHz, CDCl3): δ 211.37, 172.20, 170.94, 154.92, 135.23, 128.56, 128.41, 128.22, 79.81, 67.17, 60.53, 56.06, 53.39, 52.88, 36.79, 32.36, 30.18, 28.22, 27.00.

[0171] Chemical data for compound 21: ¹H NMR (300 MHz, CDCl3): δ 7.40-7.20 (m, 5H), 5.49 (brd, J = 7.7

Hz, 1H), 5.17 (s, 2H), 5.09 (m, 1H), 4.52 (t, J = 8.5 Hz, 1H), 4.22 (m, 1H), 3, 08 (dd, J = 12.7, 4.5 Hz, 1H), 2.92 (m, 1H), 2.60 (m, 2H), 2.36-1.72 (m, 6H), 1 , 43 (brs, 9H); ¹³C NMR (75 MHz, CDCl3): 8 207.72, 170.93, 170.15, 154.74, 135.58, 128.37, 128.30, 128.14, 80.00, 66.67, 60.10, 59.74, 52.13, 48.52, 39.65, 34.18, 32.36, 28.21, 26.90.

<img file="PL2019671T3_D0050.tif" />

[0172] To a solution of compound 20 (160 mg, 0.37 mmol) in 15 mL methanol was added NaBH3CN (120 mg, 1.9 mmol) and 3 drops of H2SO4 (98%) at -15 ° C. After stirring at the same temperature for 4 h, 10 ml of water were added and the mixture was extracted with ethyl acetate (30 ml χ 4). The combined organic layers were dried over Na2SO4 and then concentrated. The residue was purified by chromatography to give compound 22 (147 mg, 92%).

EP 2 019 671 B1 [0173] ¹H NMR (300 MHz, CDCl3): 8 7.30 (brs, 5H), 5.45 (brd, J = 8.4 Hz, 1H), 5.20, 5.10 (ABq, J = 14.1 Hz, 2H), 4.85 (m, 1H), 4.48 (m, 2H), 4.13 (m, 1H), 3.16 (brs, 1H), 2.42-1.45 (m , 10H), 1.38 (brs, 9H); ¹³C NMR (75 MHz, CDCl3): δ 172.30, 171.03, 155.06, 135.43, 128.49, 128.27, 128.15, 79.36, 67.70, 66.87, 59.74, 54.00, 51.65, 43.62, 32.12, 31.82, 29.30, 28.29, 27.11.

<img file="PL2019671T3_D0051.tif" />

[0174] To a solution of compound 19 (170 mg, 0.39 mmol) in 10 mL CH2Cl2 was added methanesulfonyl chloride (0.05 mL, 0.6 mmol). The solution was cooled to 0 ° C and then 0.2 ml of N, N-diisopropylethylamine was added dropwise. The mixture was stirred at room temperature for 4 h and then concentrated. The residue was purified by chromatography to give a mesylate. To a solution of this mesylate in 10 ml of methanol was added 50 mg of 10% Pd-C. After stirring the mixture at room temperature under H2 atmosphere for 3 h, the catalyst was filtered off and the filtrate was concentrated to give an acid. The acid was dissolved in 10 mL CH2Cl2. To this solution was added successively (R) - (-) - 1,2,3,4-tetrahydro-1-naphthylamine (60 mg, 0.4 mmol), EDC (77 mg, 0.4 mmol), HOBt (55 mg, 0.4 mmol) and 0.3 mL of N, N-diisopropylethylamine. The solution was stirred at room temperature overnight and then concentrated. The residue was purified by chromatography to give an amide. To a solution of this amide in 5 ml DMF was added 0.2 g NaN3. The mixture was stirred at 110 ° C for 6 h and then partitioned between 60 ml ethyl acetate and 15 ml brine. The organic layer was dried over Na2SO4 and then concentrated. The residue was purified by chromatography to give compound 23 (132 mg, 68% in four steps).

[0175] ¹H NMR (300 MHz, CDCl3): 8 7.30 (m, 1H), 7.20-7.03 (m, 3H), 6.81 (brd, J = 8.2 Hz, 1H), 5, 50 (brd, J = 8.4 Hz, 1H), 5.18 (m, 1H), 4.65 (m, 1H), 4.48 (m, 1H), 4.35 (m, 1H), 3.84 (m, 1H), 2.80 (m, 2H), 2.45 (m, 1H), 2.30-1.78 (m, 13H), 1.45 (brs, 9H); ¹³C NMR (75 MHz, CDCl3): δ 171.19, 170.26, 154.87, 137.18, 136.41, 129.07, 128.73, 127.28, 126.16, 79.56, 60.47, 59.30, 54.47, 50.79, 47.57, 40.40, 33.19, 32.56, 29.83, 29.02, 28.32, 27.82, 25, 43, 19.74.

<img file="PL2019671T3_D0052.tif" />

[0176] Compound 24 was synthesized in the same sequence as compound 6 from compound 22 (63% in four steps).

[0177] ¹H NMR (300 MHz, CDCl3): δ 7.32 (m, 1H), 7.15 (m, 2H), 7.06 (m, 1H), 6.77 (brd, J = 8.3 Hz, 1H), 5.43 (brd, J = 8.2 Hz, 1H), 5.16 (m, 1H), 4.54 (m, 1H), 4.48 (t, J = 7.5 Hz, 1H), 4.25 (m, 1H), 3.52 (m, 1H), 2.80 (m, 2H), 2.48-1.50 (m, 14H), 1.42 (brs, 9H ); ¹³C NMR (75 MHz, CDCl3): δ 171.02, 170.07, 154.94, 137.11, 136.33, 128.98, 128.76, 127.20, 126.23, 79.76, 61.17, 60.35, 56.99, 49.85, 47.47, 42.20, 32.74, 29.84, 29.14, 29.01, 28.25, 26.08, 19, 76.

EP 2 019 671 B1

<img file="PL2019671T3_D0053.tif" />

[0178] To a solution of 20 mg CuSO4 in 2 ml water was added 40 mg (+) - L-ascorbate sodium. The mixture was shaken until it turned bright yellow. This mixture was added dropwise to a solution of compound 23 (120 mg, 0.24 mmol) and 0.2 mL propargyl ether in 3 mL acetonitrile and 3 mL t-BuOH at room temperature. The mixture was stirred at the same temperature overnight and then partitioned between 60 ml CH2Cl2 and 15 ml brine. The organic layer was dried over Na2SO4 and then concentrated. The residue was purified by chromatography to give compound 25 (105 mg, 74%).

[0179] ¹H NMR (300 MHz, CDCl3): δ 7.64 (s, 1H), 7.39 (m, 1H), 7.18 (m, 2H), 7.07 (m, 1H), 6.50 ( brd, J = 8.3 Hz, 1H), 5.83 (brd, J = 7.1 Hz, 1H), 5.19 (m, 1H), 4.76 (m, 1H), 4.70 ( s, 2H), 4.55-4.32 (m, 3H), 4.23 (d, J = 2.4 Hz, 2H), 2.80 (m, 2H), 2.50 (t, J = 2.4 Hz, 1H), 2.46-1.63 (m, 14H), 1.48 (brs, 9H); ¹³C NMR (75 MHz,

CDCl3): δ 170.63, 170.48, 154.79, 143.58, 137.35, 136.30, 129.11, 128.59, 127.36, 126.41, 122.65, 79, 71, 79.31, 74.87, 62.96, 60.83, 59.13, 57.49, 54.65, 53.29, 47.84, 41.98, 34.53, 32.49, 31.26, 30.15, 29.10, 28.34, 25.81, 20.06.

<img file="PL2019671T3_D0054.tif" />

[0180] Compound 26 was synthesized by the same method as compound 25 from compound 24 (73% yield).

[0181] ¹H NMR (300 MHz, CDCl3): 8 7.59 (s, 1H), 7.36 (m, 1H), 7.13 (m, 2H), 7.04 (m, 1H), 6.68 ( brd, J = 8.2 Hz, 1H), 5.48 (brd, J = 8.3 Hz, 1H), 5.15 (m, 1H), 4.89 (m, 1H), 4.72 ( m, 1H), 4.70 (s, 2H), 4.56-4.35 (m, 2H), 4.14 (d, J = 2.3 Hz, 1H), 2.80 (m, 2H ), 2.58-1.55 (m, 15H), 1.43 (brs, 9H); ¹³C NMR (75 MHz, CDCl3): δ 170.83, 170.60, 155.02, 144.76, 137.22, 136.29, 129.03, 128.79, 127.31, 126.32, 120.18, 79.86, 79.18,

74,97, 62,97, 61,36, 60,02, 57,52, 57,22, 49,65, 47,76, 43-81, 32,84, 32,72, 30,50, 29,92, 29,01, 28,29,

26,56, 19,86.

<img file="PL2019671T3_D0055.tif" />

[0182] To a solution of compound 23 and 25 in 3 ml acetonitrile and 3 ml t-BuOH a mixture of 20 mg CuSO4 and 40 mg sodium (+) - L-ascorbate in 2 ml water was added. The mixture was stirred at room temperature overnight and then partitioned between 60 ml CH2Cl2 and 15 ml brine. The organic layer was dried over Na2SO4 and then concentrated. The residue was purified by chromatography to give compound 27 (79%).

EP 2 019 671 B1 [0183] ¹H NMR (300 MHz, CDCl3): δ 7.70 (s, 2H), 7.40 (m, 2H), 7.19 (m, 4H), 7.11 (m, 2H), 6.54 ( brd, J = 8.3 Hz, 2H), 5.79 (brd, J = 7.1 Hz, 2H), 5.19 (m, 2H), 4.75 (m, 2H), 4.67 ( s, 4H), 4.47 (m, 4H), 4.35 (t, J = 8.5 Hz, 2H), 2.79 (m, 4H), 2.45-1.60 (m, 28H ), 1.43 (brs, 18H); ¹³C NMR (75 MHz, CDCl3): δ 170.57, 170.48, 154.78, 143.89, 137.31, 136.32, 129.06, 128.62, 127.31, 126.39, 122.60, 79.66, 63.66, 60.80, 59.06, 54.65,

53,27, 47,80, 41,93, 34,49, 32,49, 30,13, 29,21, 29,09, 28,32, 25,78, 20,03.

<img file="PL2019671T3_D0056.tif" />

[0184] Compound 28 was synthesized in the same manner as compound 27 from compounds 24 and 26.

[0185] ¹H NMR (300 MHz, CDCl3): δ 7.61 (s, 2H), 7.38 (m, 1H), 7.20-6.99 (m, 6H), 6.60 (brd, J = 8 , 0 Hz,

2H), 5.48 (brd, J = 8.1 Hz, 2H), 5.16 (m, 2H), 4.89 (m, 2H), 4.73 (m, 2H), 4.70 ( s, 4H), 4.52-4.33 (m, 4H), 2.78 (m, 4H), 2.56-1.58 (m, 28H), 1.43 (brs, 18H); ¹³C NMR (75 MHz, CDCl3): 8 170.89, 170.51, 155.01, 137.22, 136.25, 129.03, 128.81, 127.33, 126.35, 120.21, 79.87, 63.85, 61.38, 60.03, 57.27, 49.65, 47.75,

43,86, 32,85, 32,73, 30,51, 29,21, 29,03, 28,30, 26,54, 19,84.

<img file="PL2019671T3_D0057.tif" />

[0186] To a solution of compound 27 in 5 mL of methanol was added 1 mL of HCl (4 N in 1,4-dioxane). The solution was stirred at room temperature overnight and then concentrated. The residue was suspended in 5 mL CH2Cl2. LN-methyl-N-Boc-alanine, EDC, HOBt and N, N-diisopropylethylamine were added to this mixture. The mixture was stirred at room temperature overnight and then concentrated. The residue was purified by chromatography to give an amide. To a solution of this amide in 5 mL of methanol was added 1 mL of HCl (4 N in 1,4-dioxane). The solution was stirred at room temperature overnight and then concentrated to give crude SH-172 as the HCl salt. This compound was purified by HPLC to give a pure product. The gradient ran from 75% solvent A (water containing 0.1% TFA) and 25% solvent B (acetonitrile containing 0.1% of TFA) to 55% solvent A and 45% solvent over 25 min. Analytical HPLC showed greater than 95% purity.

<img file="PL2019671T3_D0058.tif" />

[0187] ¹H NMR (300 MHz, D2O): δ 7.85 (s, 2H), 7.22-7.02 (m, 8H), 4.98-4.79 (m, 4H), 4.80-4 , 60 (m, 4H),

4.52 (s, 4H), 4.28 (t, J = 8.7 Hz, 2H), 3.82 (m, 2H), 2.72-2.50 (m, 10H), 2.48 -1.55 (m, 28H), 1.45 (d, J = 7.0 Hz, 6H); ¹³C NMR (75 MHz, D2O): δ 173.57, 171.59, 169.27, 138.30, 136.14, 129.54, 128.56, 127.82, 126.61, 124.46, 62.79, 61.98, 59.66, 57.28, 55.45, 53.24, 48.38, 41.25, 31.93, 31.32, 31.11, 29.86, 28, 81,

27,24, 20,27, 15,64.

EP 2 019 671 B1

SH-153

<img file="PL2019671T3_D0059.tif" />

SH-153 was synthesized in the same sequence as SH-172 from compound 28.

[0188] ¹H NMR (300 MHz, D2O): δ 7.95 (s, 2H), 7.20-6.95 (m, 8H), 4.92-4.74 (m, 6H), 4.59 (s , 4H), 4.55 (m,

2H), 4.27 (m, 2H), 3.86 (m, 2H), 2.78-2.52 (m, 12H), 2.42-2.08 (m, 8H), 1.99 -1.55 (m, 18H), 1.45 (d, J = 7.0 Hz, 6H); ¹³C NMR (75 MHz, D2O): δ 172.99, 171.58, 169.78, 138.23, 136.07, 129.46, 128.57, 127.78, 126.57, 123.39, 62.82, 62.56, 60.59, 58.14, 57.13, 50.49, 48.38, 42.62, 32.49, 31.30, 29.84, 29.53, 28, 78,

27,86, 20,20, 15,61.

EXAMPLE 5

Synthesis of SH-146 [0189] Compound SH-146 was synthesized according to Scheme VI.

Diagram VI:

<img file="PL2019671T3_D0060.tif" />

SH-146

Reagents and conditions: (a) i. 2 N LiOH, 1,4-dioxane: H2O 1: 1; ii. 15, EDC, HOBt, N, N-diisopropylethylamine, CH2Cl2. 88% in two stages; (b) i. 24. CuSO4, (+) - sodium L-ascorbate, tBuOH-H2O 1: 1, ii. 4N HCl in 1,4-dioxane, MeOH; iii. LN-Boc-N-methylalanine, EDC, HOBt, N, N-diisopropylethylamine, CH2Cl2; iv. 4N HCl in 1,4-dioxane, MeOH, 55% in four steps.

[0190] Removal of the trifluoroacetyl protecting group in compound 3, followed by condensation of the resulting amine with acid 15 gave amide 29. Cycloaddition of azide 24 with alkyne 29 under catalysis with CuSO4 (+) - sodium L-ascorbate, followed by removal of the Boc protecting groups gave the salt ammonium. After condensation of this salt with LN-Boc-N-methylalanine, the Boc protecting groups were cleaved by treatment with HCl in methanol to form divalent Smac SH-146 mimetics.

<img file="PL2019671T3_D0061.tif" />

EP 2 019 671 B1 [0191] ¹H NMR (300 MHz, CDCl3): δ 7.75 (brs, J = 7.4 Hz, 1H), 7.32-7.15 (m, 5H), 5.53 (brd, J = 7.1 Hz, 1H), 5.08 (m, 1H), 4.73 (t, J = 6.6 Hz, 1H), 4.60 (m, 1H), 4.19 (brs, 2H), 4.12 ( m, 1H), 3.80 (m, 2H), 2.62 (m, 1H), 2.45 (t, J = 2.3 Hz, 1H), 2.23-1.65 (m, 4H ), 1.55-1.10 (m, 16H); ¹³C NMR (75 MHz, CDCl3): δ 172.20, 169.94, 155.03, 139.45, 128.37, 127.46, 127.05, 79.59, 79.23, 74.91, 72.23, 59.76, 59.27, 58.34, 52.96, 51.08, 36.44, 32.00, 28.37, 24.93, 24.12, 23.08.

<img file="PL2019671T3_D0062.tif" />

[0192] ¹H NMR (300 MHz, D2O): δ 7.84 (p. 1H), 7.40-7.25 (m, 6H), 7.24-7.02 (m, 3H), 4.98-4 , 85 (m, 4H), 4.74 (m, 2H), 4.53 (s, 2H), 4.30 (m, 2H), 4.27 (m, 1H), 3.97-3, 80 (m, 2H), 3.78-3.65 (m, 2H), 2.92 (m, 2H), 2.56 (s, 3H), 2.55 (s, 3H), 2.35 -1.45 (m, 26H), 1.43 (d, J = 7.0 Hz, 3H), 1.38 (d, J = 7.0 Hz, 3H); ¹³C NMR (75 MHz, D2O): δ 173.88, 173.59, 172.24, 171.57, 169.52, 169.26, 143.92, 138.65, 138.31, 136.20, 129.51, 129.12, 128.48, 128.17, 127.80, 126.95, 126.60, 124.24, 72.30, 63.35, 62.21, 61.98, 61, 05, 59.61, 57.27, 57.19, 55.46, 53.35, 53.20, 51.12, 48.41, 41.21, 35.95, 33.00, 32.34, 31.30, 29.83, 28.80, 27.94, 25.08, 21.91, 20.30, 15.62.

EXAMPLE 6

Synthesis of YP-317, YP-381, YP-383 and YP-385 [0193] Compound YP-317 was synthesized according to Scheme VII.

EP 2 019 671 B1

<img file="PL2019671T3_D0063.tif" />

Reagents and conditions: (a) tert-butyldimethylsilyl chloride, N, N-diisopropylethylamine, methylene chloride; (b) 10% Pd-C, methanol, H2, 88% in two steps; (c) Boc-Dap (Z) -OH, EDC, HOBt, N, N-diisopropylethylamine, methylene chloride; (d) 1 M tetrabutylammonium fluoride in tetrahydrofuran,

Tetrahydrofuran, 87% in two stages; (e) Dess-Martin periodinane, methylene chloride, 96%; (f) 10% Pd-C, methanol, H2, 64%; (g) benzyl chloroformate, sodium bicarbonate, 1,4-dioxane, 95%; (h) thionyl chloride, methanol; (i) Boc anhydride, sodium bicarbonate, 1,4-dioxane, 71% in two steps; (j) 2 M lithium hydroxide in H2O, 1,4-dioxane-H2O; (k) aminodiphenylmethane, EDC, HOBt, N, N-diisopropylethylamine, methylene chloride, 74% in two steps; (1) 4 M hydrogen chloride in 1,4-dioxane, methanol; (m) LN-Boc-N-methylalanine, EDC, HOBt, N, N-diisopropylethylamine, methylene chloride, 78% in two steps; (n) 10% Pd-C, methanol, H₂, 90%; (o) α, α'-dibromo-p-xylene, sodium bicarbonate, 1,4-dioxane; (p) compound 21, sodium bicarbonate, 1,4-dioxane; (q) 4M hydrogen chloride in 1,4-dioxane, methanol, 34% in three stages.

[0194] Compound 31 was prepared in six steps from pyroglutamic acid (compound 30) by published methods (Zhang et al., Org. Lett., 4: 4029-4032 (2002); (b) Polyak et al. , J. Org, Chem. 63: 5937-5949 (1998)). The hydroxyl group in compound 31 was protected by TBS to give compound 32. The benzyl protecting group was removed by catalytic hydrogenation to give amine 33, which was coupled with Boc-Dap (Z) -OH to give amide 34. The TBS protecting group in compound 34 was removed by treatment with 1 M tetrabutylammonium fluoride in tetrahydrofuran to give alcohol 35, which was then oxidized to aldehyde 36 with Dess-Martin periodinate. Removal of the Cbz protecting group in compound 36 by catalytic hydrogenation, intramolecular condensation of the desired amine with aldehyde, and subsequent reduction of the resulting enamine was carried out in one pot to give bicyclic compound 37. Amine 37 was protected with a Cbz protecting group to give compound 38. The Boc protecting group in compound 38 was removed and the tert-butyl ester was converted to the methyl ester by treatment with thionyl chloride in methanol to give compound 39. The amino group was protected with a Boc protecting group to give compound 40. Methyl ester 40 was converted to carboxylic acid 41, which was condensed with aminodiphenylmethane to give amide 42. The Boc protecting group in compound 42 was removed to give free amine 43, which was coupled to LN-Boc-N-methylalanine to give amide 44. The Cbz protecting group in compound 44 was removed by catalytic hydrogenation to give amine 45, which was treated with α, α'- dibromo-p-xylene to give compound 46. Compound 46 was treated with compound 45 and sodium bicarbonate to give the protected dimer 47, whose Boc protecting groups were removed to give the desired YP-317 dimer.

[0195] ¹H NMR (D2O) δ 9.23-9.26 (m, 1H), 7.18-7.22 (m, 8H), 7.06-7.08 (m, 4H), 6.02-6 , 05 (m, 1H), 5,305,36 (m, 1H), 4.59 (m, 1H), 4.52 (m, 1H), 4.27 (s, 2H), 3.78-3 , 86 (m, 2H), 3.53-3.58 (m, 2H), 2.99 (t, J = 2 Hz, 1H), 2.56 (s, 3H), 2.32 (m, 1H), 1.84-1.92 (m, 1H), 1.67-1.75 (m, 4H), 1.35-1.37 (d, J = 7 Hz, 3H); ¹³C NMR (D2O) δ 173.93, 170.13, 167.90, 163.54, 163.07, 140.74, 140.62, 132.39, 130.59, 129.39, 129.26, 128.40, 128.13, 127.85, 127.58, 118.58, 114.72, 62.98, 61.07, 59.03, 58.25, 57.10, 56, 55, 26, 54.18,

47.98, 31.65, 31.24, 30.98, 27.01, 15.42; HRMS. Found: [M + H]⁺ 1057.6057 (calc. 1057.6028). [0196] YP-381, YP-383 and YP-385 were synthesized in a similar manner by acylation of compound 45.

<img file="PL2019671T3_D0064.tif" />

[0197] ¹H NMR (300 MHz, CD3OD, TMS) δ 8.94-8.91 (d, J = 7.9 Hz, 1H), 7.37-7.24 (m, 10H), 6.17-6, 14 (d,

J = 8.2 Hz, 1H), 4.58-4.55 (m, 1H), 4.24 (br, 1H), 3.99-3.90 (m, 2H), 3.50-3 , 38 (m, 1H), 2.70 (s, 3H), 2.652.40 (m, 2H), 2.31 (m, 1H), 2.09-1.78 (m, 6H), 1, 61 (m, 2H), 1.56-1.53 ​​(d, J = 7.0 Hz, 3H), 1.32 (s, 4H). ¹³CNMR (75 MHz, CD3OD) δ 176.5, 175.9, 173.2, 169.6, 143.0, 129.6, 128.8, 128.2, 62.7, 58.2, 53, 7, 34.4,

33.4, 32.3, 31.8, 30.3, 28.3, 26.1, 16.2. HRMS: calculated m / z for [M + Na]⁺ 1143.6371; found 1143.6387.

YP-383

EP 2 019 671 B1

<img file="PL2019671T3_D0065.tif" />

[0198] ¹H NMR (300 MHz, CD3OD, TMS) δ 7.35-7.27 (m, 10H), 6.16 (s, 1H), 4.59-4.52 (m, 1H), 4.24 ( br, 1H), 4.02-3.99 (m, 1H), 3.97-3.92 (m, 1H), 3.90-3.87 (m, 1H), 3.58-3, 48 (m, 2H), 2.70 (s, 3H), 2.56-2.34 (m, 2H), 2.34-2.32 (m, 1H), 2.06-1.19 ( m, 6H), 1.56-1.53 ​​(d, J = 7.0 Hz, 3H), 1.30 (s, 6H). ¹³CNMR (75 MHz,

CD3OD) δ 176.5, 175.9, 173.2, 169.6, 143.0, 129.6, 128.8, 128.2, 62.7, 58.2, 53.7, 34.4 , 33.4, 31.7, 30.6,

30.4, 28.3, 26.3, 16.1. HRMS: calculated m / z for [M + Na]⁺ 1171.6684; found 1171.6680.

<img file="PL2019671T3_D0066.tif" />

[0199] ¹H NMR (300 MHz, CD3OD, TMS) δ 8.95-8.93 (d, J = 8.1 Hz, 1H), 7.37-7.24 (m, 10H), 6.17-6, 14 (d, J = 8.0 Hz, 1H), 4.58-4.56 (m, 1H), 4.24 (br, 1H), 3.99-3.92 (m, 2H), 3 , 82-3.70 (m, 1H), 2.70 (s, 3H), 2.6210 2.40 (m, 2H), 2.33 (m, 1H), 2.04-1.75 ( m, 6H), 1.60-1.56 (m, 2H), 1.55-1.52 (d, J = 7.0 Hz, 3H), 1.34-1.30 (m,

2H). ¹³CNMR (75 MHz, CD3OD) δ 176.5, 175.8, 173.3, 169.7, 143.0, 129.6, 128.8, 128.2, 62.7, 58.2, 53, 7

34.5, 33.4, 31.7, 30.2, 28.3, 26.2, 16.1. HRMS: calculated m / z for [M + Na]⁺ 1115.6058; found 1115.6055.

EXAMPLE 7 [0200] Synthesis of Smac mimetics intermediates

Diagram VIII

<img file="PL2019671T3_D0067.tif" />

Reagents and conditions: (a) i. 4 N HCl in 1,4-dioxane, methanol; ii. Boc-Dap (Z) -OH, EDC, HOBt, N, N-diisopropylethylamine, CH 2 Cl 2, 52% in two steps; (b) O3, then PPh3, CH2Cl2, 90%; (c) H2, 10% Pd-C, i-PrOH, 41%; (d) 9-BBN (2 equivalents), THF, reflux, 12 h, then 3 N NaOH (2 equivalents), 35% H2O2 (2.5 equivalents), 0 ° C - room temperature, 85%; (e) Dess-Martin periodinane, CH2Cl2; ii.

H2, 10% Pd-C, i-PrOH, 50% in two stages.

[0201] The synthesis of intermediates 51 and 53 is outlined in Scheme VIII. Compound 48 can be produced in five steps from pyroglutamic acid according to the described methods (see: (1) Zhang, J .;

EP 2 019 671 B1

Xiong, C .; Wang, W .; Ying, J .; Hruby, V., J. Org. Lett., 2002, 4 (23), 4029-4032 and (2) Polyak, F. and Lubell, WDJ Org, Chem. 1998, 63, 5937-5949) as a mixture of two diastereomers with the R isomer as the main product (the ratio is about 4: 1). Removal of the Boc group at 48 followed by condensation with N-α (te / t-butoxycarbonyl) -Ne- (benzoxycarbonyl) -L-diamino-propionic acid (Boc-Dap (Z) -OH) gave amide 49. Double oxidation with ozone CC binding at 49 gave 50 aldehyde. Cleavage of the Cbz group into 4, intramolecular condensation of the resulting amine with an aldehyde group and subsequent reduction of the enamine were carried out in one pot to form compound 51. In this conversion only compound 51 was obtained and no detectable formation of its isomer was found, suggesting that the aminoaldehyde from the minor isomer cyclizes under these conditions.

[0202] Hydroboration of the double CC bond in 49 9-BBN followed by basic oxidation of the resulting borane gave alcohols 52. Oxidation with Dess-Martin periodinane gave a mixture of two aldehydes that was cyclized in the same procedure as for compound 51 to give compound 53. Similarly as for 51, only one isomer was obtained during this conversion.

[0203] Analytical data for compound 51: [α]²% - 30.2 (c = 1.7, CHCl3); ¹H NMR (300 MHz, CDCL TMS) δ

5.45 (brd, J = 8.0 Hz, 1H), 4.67 (m, 1H), 4.52 (t, J = 9.0 Hz, 1H), 4.23 (m, 1H), 3.74 (s, 3H), 3.20 (m, 2H), 2.94 (m, 1H), 2.74 (dd, J = 13.6, 10.9 Hz, 1), 2.35 (m, 1H), 2.14 (m, 1H), 1.99 (m, 1H), 1.86-1.74 (m, 3H), 1.66 (m, 1H), 1.43 ( brs, 9H); ¹³C NMR (75 MHz, CDCl3, TMS) δ 173.42, 170.60, 155.16, 79.68, 59.46, 58.39,

54,92, 52,44, 46,72, 37,45, 32,15, 29,64, 28,29, 26,98.

[0204] Analytical data for compound 53: [α]²% - 23.2 (c = 1.0, CHCl3); ¹H NMR (300 MHz, CDCl3, TMS) δ 5.23 (brd, J = 8.0 Hz, 1H), 4.79 (m, 1H), 4.65 (dd, J = 9.7, 8, 2 Hz), 4.22 (m, 1H), 3.74 (s, 3H), 3.02-2.80 (m, 4H), 2.38-1.70 (m, 9H), 1, 43 (brs, 9H); ¹³C NMR (75 MHz, CDCl3, TMS) δ 173.38, 171.59, 155.09, 79.68, 62.03, 59.82, 53.72, 53.15, 52.48, 50.09 , 34.66, 34.55, 29.47, 28.31, 27.33.

EXAMPLE 8

Synthesis of SH-188, 189 and 190 [0205]

Diagram IX

<img file="PL2019671T3_D0068.tif" />

Reagents and conditions: (a) EDC, HOBt, NN-diisopropylethylamine, CH2Cl2; (b) i. 4 N HCl in 1,4-dioxane, MeOH; ii. (S) -N-Boc-N-methylalanine, EDC, HOBt, N, N-diisopropylethylamine, CH2Cl2; iii. 10% Pd-C, H2, MeOH; (c) thiophosgene or triphosgene, CH2Cl2; ii. 4N HCl in 1,4-dioxane, MeOH.

[0206] Condensation of acid 15 with amine 54 or 55 gave amides 56 and 57, respectively (Scheme IX). Removal of the Boc protecting group at 56 or 57 followed by condensation of the resulting (S) -NBoc-N-methylalanine ammonium compounds gave two amides. Removal of the Cbz protecting groups in these two amides gave amines 58 and 59. Condensation 58 or 59 with 0.5 equivalent of triphosgene gave two ureas. Removal of Boc protecting groups in these two ureas provided SH-188 and SH-190, respectively. Condensation 58 of 0.5 equivalent of thiophosgene gave thiourea. Removal of the Boc protecting group in this thiourea gave SH-189.

EXAMPLE 9

SH-202 Synthesis [0207]

Scheme X

<img file="PL2019671T3_D0069.tif" />

Reagents and conditions: (a). i. NaH (2.5 equivalents), 1,12-dibromododecane, DMF; ii. 3 N LiOH, 1,4-dioxane; (b) acid (2.2 eq), EDC, HOBt, N, N-diisopropylethylamine, CH 2 Cl 2; (c) i. 4 N

HCl, 1,4-dioxane, MeOH, ii. (S) -N-Boc-N-methylalanine, EDC, HOBt, N, N-diisopropylethylamine,

CH2Cl2; iii. 4 N HCl, 1,4-dioxane, MeOH.

[0208] Substitution of sodium alkoxide derived from compound 60 with 1,12-dibromododecane followed by removal of the trifluoroacetyl group gave diamine 61. Condensation 61 with 2 equivalents of acid gave amide 62. Removal of Boc protecting groups at 62 followed by condensation with (S) -N-Boc-N-methylalanine gave an amide. Removal of the Boc protecting group in this amide provided SH-202.

EXAMPLE 10

Synthesis of Smac mimetic intermediates [0209]

Diagram XI

<img file="PL2019671T3_D0070.tif" />

Reagents and conditions: (a) i. 4 N HCl in 1,4-dioxane, methanol; ii. Boc-Dap (Z) -OH, EDC, HOBt, N, N-diisopropylethylamine, CH 2 Cl 2; (b) O3, then PPh3, CH2Cl2; (c) H2. 10% Pd-C, i-PrOH; (d) i. CbzCl,

EP 2 019 671 B1

NaHCO3, 1,4-dioxane; ii. 3 N LiOH, 1,4-dioxane then 1 N HCl; (e) i. amine, EDC, HOBt, N.Ndiisopropylethylamine, CH2Cl2; ii. 4 N HCl in 1,4-dioxane, methanol: iii. (S) -N-protected amino acid, EDC, HOBt, N, N-diisopropylethylamine, CH2Cl2; (f) i. diacid (0.5 eq), EDC, HOBt, N, N-diisopropylethylamine, CH 2 Cl 2; ii. 4 N HCl in 1,4-dioxane, methanol.

[0210] A new and efficient method for the synthesis of key intermediate 41 is shown in Scheme XI. Compound 48 can be produced in five steps from pyroglutamic acid according to the described methods (see: (1) Zhang, J .; Xiong, C .; Wang, W .; Ying, J. and Hruby, V., J. Org. Lett ., 2002, 4 (23), 4029-4032 and (2) Polyak, F. and Lubell, WDJ Org, Chem. 1998, 63, 5937-5949) as a mixture of two diostereoisomers with the R isomer as the main product (the ratio is about 4: 1). Removal of the Boc group in 48 followed by condensation with W-α- (ίerί-butoxycarbonyl) -W-β- (benzoxycarbonyl) -L-diamino-propionic acid (BocDap (Z) -OH) acid gave amides 49. Double bond ozone oxidation CC at 49 followed by reduction of PPh3 gave an aldehyde of 50. The cleavage of the Cbz group at 50, intramolecular condensation of the resulting amine with the aldehyde group and subsequent reduction of the enamine were done in one pot to give the desired compound 51. Protection of the amino group followed by hydrolysis of the methyl ester at 51 gave acid 41.

[0211] Condensation 41 with various diamines gave a series of amides. Removal of the Boc protecting groups in these amides followed by condensation of the resulting ammonium salts with (S) -N-protected amino acids gave a series of amides 63. Removal of the protecting groups in these amides gave Smac 64 divalent target mimetics.

EXAMPLE 11

DIVAL VALUE SMAC MIMETICS [0212]

<img file="PL2019671T3_D0071.tif" />

SH-173 [0213] SH-173: ¹H NMR (300 HMz, CDCl3): δ 8.70 (s, 2H), 8.32 (d, J = 8.0 Hz, 2H), 7.60 (d, J = 7.4 Hz, 2H) .

7.40-6.80 (m, 26H), 6.47 (d, J = 7.9 Hz, 2H), 6.25 (d, J = 8.0 Hz, 2H), 4.81 ( m, 2H), 4.70 (m, 2H), 4.50 (m, 2H), 4.27 (t, J = 7.1 Hz, 4H), 3.95 (m, 2H), 3, 30-3.08 (m, 4H), 2.60 (t, J = 2.61 Hz, 4H), 2.45-2.25 (m, 2H), 2.12-1.20 (m, 36H); ¹³C NMR (75 HMz, CDCl3): δ 170.78, 170.41, 170.14, 169.88, 148.00, 140.45, 138.98, 136.19, 128.53, 128.44, 127.68, 127.56, 127.28, 123.70, 121.67, 121.55, 119.13, 118.57, 111.29, 110.25,

59,98, 59,06, 54,01, 50,40, 50,20, 50,01, 47,76, 35,93, 34,61, 31,74, 29,53, 29,09, 28,08, 24,83, 23,30,

22,95, 20,82;

<img file="PL2019671T3_D0072.tif" />

SH-175 [0214] SH-175: ¹H NMR (300 HMz, D2O): δ 7.87 (s, 1H), 7.71 (s, 2H), 7.30-7.11 (m, 10H), 6.05 (s, 2H), 4.64 (m, 2H), 4.35-4.16 (m, 10H), 3.81 (m, 2H), 2.60 (t, J = 6.2 Hz, 2H), 2.54 (s, 6H), 2.10 (m, 2H), 1.98 (m, 2H), 1.75-1.42 (m, 34H); ¹³C NMR (75 HMz, D2O): δ 175.85, 174.79, 172.08, 150.63, 147.37, 141.60, 131.85, 131.09, 129.93, 128.50, 126.79, 64.57, 63.53, 59.73, 54.54, 53.66, 52.90, 52.59, 38.49, 35.57, 34.90, 33.93,

31,29, 30,32, 28,96, 28,65, 27,63, 27,38, 25,24, 24,49, 18,22.

EP 2 019 671 B1

<img file="PL2019671T3_D0073.tif" />

[0215] SH-176: ¹H NMR (300 MHz, D2O): δ 7.48 (s, 2H), 7.10-6.67 (20H), 5.84 (s, 2H), 4.65 (m, 2H), 4, 55 (m, 2H), 4.42-4.16 (m, 8H), 3.80 (m, 2H), 2.53 (s, 6H), 2.25-1.30 (m, 26H) ; ¹³C NMR (75 MHz, D2O): δ 172.72, 171.00, 169.10, 143.67, 141.49, 141.30, 129.07, 127.96, 127.58, 127.39, 124.10.62,76.61.24, 59.43, 57.67, 57.27, 55.00, 53.20, 41.26, 32.12, 31.31, 26.98, 24, 69, 15.62.

<img file="PL2019671T3_D0074.tif" />

[0216] SH-177: ¹H NMR (300 MHz, D2O): δ 7.62 (s, 2H), 7.15-6.90 (20H), 5.90 (s, 2H), 4.75 (m, 4H), 4, 43 (s, 4H), 4.39 (m, 4H), 3.83 (m, 2H), 2.80 (s, 6H), 2.46 (m, 2H), 2.22-1.90 (m, 8H), 1.75-1.43 (m, 10H), 1.36 (d, J = 8.4 Hz, 6H); ¹³C NMR (75 MHz, D2O): δ 175.14, 173.74, 172.25, 146.86, 144.06, 143.66, 131.72, 131.66, 130.55, 130.21, 129.97, 125.60, 65.46, 64.63, 62.98, 60.36, 59.70, 52.87, 45.02, 34.98, 33.87,

32,30, 30,26, 18,17.

<img file="PL2019671T3_D0075.tif" />

[0217] SH-178: ¹H NMR (300 MHz, D2O): δ 7.88 (s, 2H), 7.46-7.30 (m, 10H), 6.19 (s, 2H), 4.78 (m, 6H), 4.42 (m, 2H), 4.30 (m, 2H), 3.90 (m, 2H), 3.74 (m, 4H), 3.60 (m, 4H), 3.52 (m , 2H), 3.28 (m, 2H), 2.64 (s, 6H), 2.40-1.56 (m, 24H), 1.49 (d, J = 7.0 Hz, 6H) ; ¹³C NMR (75 MHz, D2O): δ 176.14, 174.90, 172.10, 151.28, 141.51, 131.90, 131.19, 130.01, 127.32, 64.64, 63.61, 59.73, 58.06, 53.68, 52.96, 52.05, 47.49, 38.50, 35.53, 34.90, 33.89, 30.35, 27, 61, 24.48, 18.18.

<img file="PL2019671T3_D0076.tif" />

[0218] SH-179: ¹H NMR (300 MHz, D2O): δ 7.87 (s, 2H), 7.40-7.22 (m, 10H), 6.19 (s, 2H), 4.74 (m, 6H), 4.42 (m, 2H), 4.30 (m, 2H), 4.05 (m, 2H), 3.79 (t, J = 5.1 Hz, 4H), 3.71 (t, J = 5.1 Hz, 4H), 3.53 (brs, 8H), 3.10 (m, 4H), 2.40-1.60 (m, 24H), 1.50 (d, J = 7, 0 Hz, 6H); ¹³C NMR (75 MHz, D2O): δ 176.15, 174.92, 172.00, 151.25, 141-51, 131.90, 131.19, 130.07, 127.28, 64.64, 63.60, 59.57, 58.60, 58.06, 53.69, 52.95, 52.14, 50.67, 47.56, 38.49, 35.53, 34.89, 30, 34, 27.60, 24.47, 18.46.

<img file="PL2019671T3_D0077.tif" />

[0219] SH-180: ¹H NMR (300 MHz, D2O): δ 7.60 (s, 2H), 7.30-7.10 (m, 10H), 6.68 (s, 4H), 6.15 (s, 2H), 4.74 (m, 2H), 4.52-4.30 (m, 4H), 4.20 (m, 2H), 4.19-4.02 (m, 8H), 3.80 (m, 2H), 3.13 (m, 2H), 2.40-1.12 (m, 42H); ¹³C NMR (75 MHz, D2O): δ 172.79, 170.36, 169.39, 148.33, 139.54, 129.24, 128.55, 127.41, 123.46, 61.91, 60.88, 60.28, 57.02, 56.06, 50.99, 50.38, 49.08, 48.13, 35.98, 34.26, 33.10, 32.32, 29, 20, 27.85, 25.11, 21.95, 16.84.

EP 2 019 671 B1

<img file="PL2019671T3_D0078.tif" />

[0220] SH-181: ¹H NMR (300 MHz, D2O): δ 7.87 (s, 2H), 7.31 (m, 4H), 7.08 (m, 4H), 6.16 (s, 2H), 4.82- 4.72 (m, 6H), 4.50 (m, 2H), 4.26 (m, 2H), 3.88 (m, 2H), 3.78 (m, 4H), 3.68 (brs , 8H), 2.61 (s, 6H), 2.36-1.53 ​​(m, 24H), 1.50 (t, J = 7.0 Hz, 6H); ¹³C NMR (75 MHz, D2O): δ 176.03, 174.85, 172.05, 163.45, 151.16, 137.40, 131.90, 127.33, 118.51, 64.59, 63.58, 59.73, 58.02, 53.64, 52.30, 51.87, 47.26, 38.48, 35.53, 34.86, 33.85, 30.27, 27, 60, 24.47, 18.16.

<img file="PL2019671T3_D0079.tif" />

[0221] SH-182: ¹H NMR (300 MHz, D2O): δ 7.61 (s, 2H), 7.23 (m, 4H), 6.78 (m, 4H), 6.53 (m, 4H), 6.12 ( s, 2H), 4.70 (m, 2H), 4.32 (m, 2H), 4.18 (m, 2H), 4.10-3.83 (m, 6H), 2.61 (s , 6H), 2.22-1.03 (m, 42H); ¹³C NMR (75 MHz, D2O): δ 172.53, 171.92, 169.39, 160.60, 148.31, 139.38, 134.84, 129.40, 128.42, 115.60, 61.76, 60.78, 57.20, 50.94, 50.27, 49.59, 35.99, 34.42, 33.22, 32.32, 31.33, 29.41, 27, 95, 25,14,21,99, 15.66.

<img file="PL2019671T3_D0080.tif" />

[0222] SH-183: ¹H NMR (300 HMz, D2O): δ 7.87 (s, 2H), 7.38-7.22 (m, 4H), 7.12-6.99 (m, 4H), 6.17 (s , 2H), 4.85-4.74 (m, 6H), 4.34 (m, 2H), 4.27 (m, 2H), 3.98 (m, 2H), 3.80-3, 65 (m, 8H), 3.55 (s, 8H), 3.10 (m, 2H),

<img file="PL2019671T3_D0081.tif" />

[0223] SH-184: ¹H NMR (300 MHz, D2O): δ 7.62 (s, 2H), 7.20 (m, 4H), 6.90-6.70 (m, 4H), 6.69-6.50 (brs , 4H), 6.19 (s, 2H), 4.72 (m, 2H), 4.50-4.28 (m, 4H), 4.20-3.80 (m, 6H), 3, 12 (m, 4H), 2.22-0.98 (m, 42H).

<img file="PL2019671T3_D0082.tif" />

[0224] SH-185: ¹H NMR (300 HMz, D2O): δ 7.67 (s, 2H), 7.26-7.10 (m, 10H), 6.07 (s, 2H), 4.73 (m, 2H), 4.28 (m, 2H), 4.25-4.10 (m, 6H), 3.83 (m, 2H), 2.54 (s, 6H), 2.20-1.92 (m, 4H), 1.86-1.30 (m, 34H), 0.98-0.80 (m, 12H); ¹³C NMR (75 HMz, D2O): δ 177.43, 173.05, 172.22, 148.26, 139.29, 129.28, 128.50, 127.43, 123.81, 62.00, 60.99, 57.20, 51.08, 50.76, 50.40, 36.00, 33.06, 32.36, 31.32, 29.52, 28.62, 28.26, 27, 77, 25.76, 25.10, 21.93, 15.65.

<img file="PL2019671T3_D0083.tif" />

[0225] SH-186: ¹H NMR (300 HMz, D2O): δ 7.58 (s, 2H), 7.20-6.90 (m, 10H), 6.61 (s, 4H), 6.08 (s, 2H), 4.72 (m, 2H), 4.30 (m, 2H), 4.10 (m, 2H), 3.95 (m, 4H), 3.80 (m, 2H), 2.52 (s , 6H), 2.25-1.05 (m, 36H), 1.02-0.75

EP 2 019 671 B1 (m, 14H); ¹³C NMR (75 HMz, D2O): δ 174.88, 174.40, 171.91, 151.04, 142.52, 142.31, 131.63, 130.92, 130.00, 125.57, 125.09, 64.30, 63.24, 59.74, 53.42, 52.90, 38.63, 38.20, 35.90, 34.85, 34.28, 33.87, 32, 69, 32.05, 31.73, 30.20, 29.07, 27.76, 24.49, 18.23.

<img file="PL2019671T3_D0084.tif" />

[0226] SH-187: ¹H NMR (300 HMz, D2O): δ 7.56 (s, 2H), 7.40-7.05 (m, 16H), 7.02 (m, 2H), 6.90 (m, 2H), 6.78 (m, 4H), 6.11 (s, 2H), 4.75 (m, 2H), 4.49 (m, 2H), 4.25 (m, 4H), 4.20 (m , 2H), 3.98 (m, 2H), 3.84 (m, 2H), 2.49 (s, 6H), 2.38 (m, 4H), 2.24-1.22 (m, 36H).

<img file="PL2019671T3_D0085.tif" />

[0227] SH-188: ¹H NMR (300 HMz, D2O): δ 7.10-6.92 (m, 10H), 6.85 (m, 4H), 6.70 (m, 4H), 5.85 (s, 2H), 4.65 (m, 2H), 4.32 (m, 2H), 4.06 (m, 2H), 3.82 (m, 2H), 2.74 (m, 4H), 2.54 (s , 6H), 2.15 (m, 4H), 2.02-1.20 (m, 34H); ¹³C NMR (75 HMz, D2O): δ 172.36, 171.92, 169.41, 160.17, 141.40, 141.15, 139.27, 129.01, 127.66, 127.46, 61.88, 60.87, 57.18, 50.94, 39.72, 35.99, 33.20, 32.49, 31.60, 31.33, 27.79, 25.19, 21, 91,

15,69.

<img file="PL2019671T3_D0086.tif" />

[0228] SH-189: ¹H NMR (300 HMz, D2O): δ 7.18-6.99 (m, 10H), 6.95 (m, 4H), 6.80 (m, 4H), 5.90 (s, 2H), 4.72 (m, 2H), 4.36 (m, 2H), 4.13 (m, 2H), 3.89 (m, 2H), 3.30 (brm, 4H), 2.63 (s , 6H), 2.30 (m, 4H), 2.12-1.18 (m, 34H).

<img file="PL2019671T3_D0087.tif" />

[0229] SH-190: ¹H NMR (300 HMz, D2O): δ ¹³C NMR (75 HMz, D2O): δ 7.15-6.93 (m, 10H), 6.95 (m, 4H), 6.79 (m, 4H), 6.85 (s, 2H), 4.74 (m, 2H), 4.36 (m, 2H), 4.13 (m, 2H), 3.88 (m, 2H), 2.80 (m, 4H), 2.58 (s , 6H), 2.36-1.08 (m, 42H).

<img file="PL2019671T3_D0088.tif" />

[0230] SH-191: ¹H NMR (300 HMz, D2O): δ 7.75 (s, 2H), 7.40-7.20 (m, 10H), 6.16 (s, 2H), 4.74 (m, 2H), 4.36 (m, 2H), 4.32-4.20 (m, 6H), 3.89 (m, 2H), 2.95 (t, J = 6.6 Hz, 4H), 2.64 (s, 6H), 2.32-1.20 (m, 38H).

<img file="PL2019671T3_D0089.tif" />

[0231] SH-198: ¹H NMR (300 HMz, D2O): δ 7.70 (s, 2H), 7.30-7.12 (m, 10H), 6.07 (s, 2H), 4.65 (m, 2H), 4.32 (m, 2H), 4.25-4.10 (m, 6H), 3.84 (m, 2H), 3.10 (m, 4H), 2.55 (s, 6H), 2 , 20-1.20 (m, 38H), 0.98 (m, 2H); ¹³C NMR (75 HMz, D2O): δ 173.17, 172.22, 169.53, 148.15, 139.16, 129.32, 128.54, 127.40, 124.04, 70.20, 69.63, 62.01, 60.98, 57.19, 51.10, 50.82, 50.63, 50.35, 35.97, 33.06, 32.36, 31.37, 29, 29, 28.17, 27.77, 26.48, 25.80, 25.10, 22.58, 21.95, 15.67.

EP 2 019 671 B1

<img file="PL2019671T3_D0090.tif" />

[0232] SH-199: ¹H NMR (300 HMz, D2O): δ 7.73 (s, 2H), 7.20-7.02 (m, 10H), 6.05 (s, 2H), 4.65 (m, 2H), 4.30 (m, 2H), 4.22-4.08 (m, 6H), 3.84 (m, 2H), 3.08 (m, 4H), 2.52 (s, 6H), 2 , 25-0.90 (m, 42H); ¹³C NMR (75 HMz, D2O): δ 173.02, 172.13, 169.49, 147.91, 139.07, 129.31, 128.55, 127.41, 124.14, 70.25, 66.87, 61.93, 60.90, 57.18, 50.97, 50.22, 36.01, 33.08, 32.37, 31.40, 29.33, 28.23, 27, 74, 25.11, 22.62.21.97, 15.70.

Ht "

<img file="PL2019671T3_D0091.tif" />

?^h NHW

Λκ n = n

Ph o

-νΎκ N'N H

SH-200 [0233] SH-200: ¹H NMR (300 HMz, D2O): δ 7.77 (s, 2H), 7.22-7.08 (m, 10H), 6.05 (s, 2H), 4.65 (m, 2H), 4.37-4.22 (m, 6H), 4.16 (m, 2H), 3.82 (m, 2H), 3.48 (m, 4H), 3.08 (m, 4H), 2 , 52 (s, 6H), 2.16-1.42 (m, 30H), 1.01 (m, 4H); ¹³C NMR (75 HMz, D2O): δ 173.11, 172.16, 169.49, 148.01, 139.10, 129.32, 128.56, 127.38, 124.62, 70.59, 68.39, 66.87, 61.96, 60.95, 57.19, 50.73, 50.24, 35.98, 33.09, 32.36, 31.40, 27.77, 25, 43, 25.12, 21.96,15,69.

<img file="PL2019671T3_D0092.tif" />

[0234] SH-201: ¹H NMR (300 HMz, D2O): δ 7.60 (s, 2H), 7.40-6.70 (m, 20), 6.16 (s, 2H), 4.75 (m, 2H), 4.49 (m, 2H), 4.25 (m, 4H), 4.20 (m, 2H), 3.98 (m, 2H), 3.84 (m, 2H), 2.49 (s , 6H), 2.40-1.20 (m , 48H).

X

<img file="PL2019671T3_D0093.tif" />

[0235] SH-202: ¹H NMR (300 HMz, D2O): δ 7.35-7.16 (m, 10H), 5.02 (m, 2H), 4.74 (m, 2H), 4.39 (m, 2H), 4.27 (m, 2H), 3.92 (m, 2H), 3.55 (m, 4H), 3.34 (m, 4H), 2.64 (s, 6H), 2.28-1 , 20 (m, 54H); ¹³C NMR (75 HMz, D2O): δ 175.35, 174.62, 172.04, 141.98, 131.60, 130.66, 129.70, 75.84, 73.59, 64.68, 63.33, 59.74, 55.57, 53.55, 38.93, 35.65, 35.05, 33.93, 32.55, 32.31, 30.64, 28.86, 27, 85, 24.68, 18.29.

<img file="PL2019671T3_D0094.tif" />

[0236] SH-206: ¹H NMR (300 HMz, D2O): δ 7.44 (s, 2H), 7.30-6.80 (m, 10H), 6.49 (s, 4H), 5.99 (s, 2H), 4.63 (m, 2H), 4.28 (m, 2H), 4.06 (m, 2H), 3.92 (m, 2H), 3.80 (m, 4H), 2.55 (s , 6H), 2.28-0.95 (m, 42H); ¹³C NMR (75 HMz, D2O): δ 175.04, 174.33, 171.97, 150.65, 143.08, 141.97, 131.63, 131.00, 130.59, 129.60, 126.25,

64,31, 63,33, 59,74, 53,47, 52,92, 38,47, 36,87, 35,85, 34,86, 33,87, 31,85, 30,42, 27,65, 24,52, 18,26.

<img file="PL2019671T3_D0095.tif" />

EP 2 019 671 B1 [0237] SM-410: ¹H NMR (MeOH-d4, 300 M Hz) δ 8.91 (m, 2H), 7.37-7.13 (m, 24H), 6.16 (m, 2H), 4.73 (m, 2H ), 4.53 (m, 2H), 4.06-3.73 (m, 8H), 3.37-3.27 (m, 6H), 2.92 (m, 6H), 2.68 ( m, 6H), 2.30 (m, 2H), 2.05-1.81 (m, 10H), 1.55 (m, 6H); ¹³C NMR (MeOH-d4, 300 M Hz) δ 174.4, 172.3, 169.3, 168.6, 142.2, 142.0, 139.3, 129.1, 128.7, 128, 5, 127.8, 127.5, 127.3, 61.8, 57.3, 52.6, 51.8, 46.6, 34.9, 32.4, 31.4, 30.9, 27.3, 15.3.

EXAMPLE 12

Binding of inhibitors to BIR3 XIAP [0238] To test the binding capacity of divalent Smac mimetics to IAP proteins, a sensitive and quantitative in vitro binding assay was developed using a method of polarized fluorescence light (FP) and used to determine the binding affinity of Smac mimetics to protein XIAP (NikolovskaColeska et al., Anal Biochem. 332: 261-73 (2004)). In this assay, 5-carboxyfluorescein (5-Fam) was coupled to the lysine side chain of the mutated Smac peptide, AbuRPF-K- (5-Fam) -NH2 (named SM5F). The kd value of SM5F peptide binding to the XIAP BIR3 protein was determined to be 17.92 nM, showing that this peptide binds to the surface pocket of the XIAP protein with high affinity. Recombinant human XIAP BIR3 XIAP protein (residues 241-356) fused with a His marker were stable and soluble, and was used for the FP-based binding assay.

[0239] Dose-dependent binding experiments were carried out with serial dilutions of test compounds in DMSO. 5 μΐ sample from test samples and pre-incubated XIAP BIR3 protein (30 nM) and SM5F peptide (5 nM) in assay buffer (100 mM potassium phosphate, pH 7.5; 100 μg / ml bovine gammaglobulin; 0.02% sodium azide , purchased from Invitrogen ™ Life Technology), were placed in 96-well, black, round-bottom Dynex plates (Fisher Scientific) for a final volume of 125 pi. Bound control peptide containing XIAP and SM5F recombinant BIR3 protein (equivalent to 0% inhibition) and free control peptide containing only free SM5F (equivalent to 100% inhibition) were included in each assay. Polarization values ​​were measured after 3 h incubation when binding reached equilibrium using ULTRA READER (Tecan US Inc., Research Triangle Park, NC). IC50 values, the concentration of the inhibitor at which 50% of the bound peptide is displaced, were determined from the plot using non-linear least squares analysis. Curve fitting was performed using GRAPHPAD PRISM software (GraphPad Software, Inc., San Diego, CA).

EXAMPLE 13

Polarized fluorescence light-based XIAP protein binding assay [0240] Smac mimetic was incubated with human XIAP (residues 120-356) (10 nM) and with Smac-based bivalent peptide with a fluorescent marker, called Smac2-F (0.5 nM) ) as a marker in assay buffer (100 mM potassium phosphate, pH 7.5; 100 ug / ml bovine gammaglobulin; 0.02% sodium azide) in 96 well black round bottom Dynex plates (Fisher Scientific). Smac2-F was determined to bind XIAP with a Kd value of 1.2 nM. For each test, controls included XIAP and Smac2-F peptide (equivalent to 0% inhibition), and only Smac-2F (equal to 100% inhibition). Polarization values ​​were measured after 2 hours of incubation using an Ultra plate reader. The IC50 value, the concentration of the inhibitor at which 50% of the bound label is eliminated, is determined from the plot using non-linear least squares analysis. Curve fitting is performed using GraphPad Prism® software.

EP 2 019 671 B1

<img file="PL2019671T3_D0096.tif" />

[0241] When tested in the binding assay, the bivalent Smac SH-164 mimetic had an IC 50 of 1.9 ± 0.5 nM (FIG. 1). This was over 500 times more than the binding affinity of the monovalent Smac SH-122 mimetic and&gt; 5000 times more potent than the natural Smac AVPI peptide (SEQ ID NO: 1). These data suggest that bivalent Smac mimetics will act as potent inhibitors of IAP activity.

EXAMPLE 14

Inhibition of cell growth by bivalent Smac mimetics [0242] The effect of SH-164 on the growth of various cancer cell lines was investigated. Cells were seeded in 96 well flat bottom culture plates at a density of 3000 cells / well with the test compound and incubated at 37 ° C in 95% air and 5% CO2 for 4 days. The rate of inhibition of cell growth after treatment with different concentrations of compound was determined using the WST-8 kit (2- (2-methoxy-4-nitrophenyl) -3- (4-nitrophenyl) -5- (2,4-disulfophenyl) -2H- monosodium salt tetrazolium; Dojindo Molecular Technologies, Inc., Gaithersburg, Maryland). WST-8 was added at a final concentration of 10% to each well, and then the plates were incubated at 37 ° C for 2-3 hours. The absorbance of the samples was measured at 450 nm using an ULTRA Tecan Reader (Molecular Device). The concentration of test compound inhibiting cell growth by 50% (IC50) was calculated by comparing absorbance in untreated cells and cells treated with the test compound.

[0243] In the assay against the human breast cancer cell line MDA-MB-231 and the MAMEL-3M melanoma cell line, SH-164 showed an IC50 of 1.4 nM (FIG. 2). In addition, SH-164 is also a potent inhibitor in several other cancer cell lines (FIG. 2).

EXAMPLE 15

Induction of cell death by Smac divalent mimetics [0244] The ability of SH-164 to induce cell death in various cancer cell lines was tested using a trypan blue viability assay. 0.3 x 10⁶ cells were seeded in 6-well plates and incubated at 37 ° C in 95% air and 5% CO2 without or with the test compound for 2 days. A 1: 1 dilution of 0.4% Trypan Blue (Invitrogen Corporation) was used to determine cell viability. SH-164 proved to be an effective inducer of cell death when incubating MDA-MB-231, MAMLE-3M and OVCAR-4 cells (FIG. 3). In each case, SH-164 induced at least 70% of cell death at a concentration of 100 nM.

EXAMPLE 16

Effect of a combination of divalent Smac mimetics and other agents on inhibiting cell growth [0245] To test the ability of divalent Smac mimetics to sensitize cancer cells to growth inhibitory effects of other agents, cell growth inhibition tests were performed with various agents alone or in combination with increasing doses of divalent Smac mimetics. . Exposure of MDA-MB-231 (2LMP) breast cancer cells to TRAIL alone resulted in an IC50 of 2.2 ng / ml (FIG.

EP 2 019 671 B1

4A). The combination of TRAIL and SH-164 significantly reduced the IC 50 for TRAIL, where TRAIL in the presence of 100 nM SH164 showed an IC 50 0.008 ng / ml. A similar result was observed for MDA-MB-453 breast cancer cells, where TRAIL alone had an IC50&gt; 1000 ng / ml and the combination of TRAIL and 100 nM SH-164 had an IC50 of 16 ng / ml (FIG. 4B). When PC-3 human prostate cancer cells were used, TRAIL alone had an IC50&gt; 300 ng / ml, whereas

TRAIL in the presence of 100 nM SH-164 had an IC 50 of 2 ng / ml (FIG. 4C). SH-122 had a similar, although weaker effect, where the combination of TRAIL and 1000 nM SH-122 had an IC50 of 30 ng / ml. In contrast, 500 nM SH-149 did not reduce TRAIL IC50.

[0246] SH-164 has also been tested for its ability to increase the growth inhibitory effect of chemotherapeutic agents, cisplatin and mitoxantrone. When tested on human breast cancer cells

MDA-MB-231 (2LMP), SH-164 at a concentration of 100 nM sensitized cells to growth inhibition by both agents (FIG. 5). These data indicate that Smac divalent mimetics are capable of sensitizing cells to growth inhibitory activities for various cancer treatment agents.

EXAMPLE 17

Apoptosis assay [0247] Apoptosis analysis was performed using an apoptosis detection kit (BioVision Research

Products, Mountain View, CA) according to the manufacturer's protocol. Briefly, cells were treated with Smac mimetics for 12 hours, harvested and washed with ice-cold PBS. Cells were stained with annexin V-FITC and propidium iodide (PI) for 15 minutes at room temperature in the dark and immediately analyzed by a FACS Calibur flow cytometer (Becton Dickinson, Erembodegem, Belgium). Stained cells

Annexin V (+) and PI (-) were considered to be apoptotic cells at an early stage. Annexin V (+) and PI (+) stained cells were considered late stage apoptotic and Annexin V () and PI (+) stained cells were considered necrotic (FIG. 6).

EXAMPLE 18 [0248] Binding affinities for XIAP and activities for inhibiting cell growth [0249] Binding affinities for XIAP and activities for inhibiting cell growth of Smac mimetics is shown in Table 5.

Table 5

<td>Relationship</td><td>IC values₅₀ bindings with XIAP [nM]</td><td>IC50 values ​​(nM) in the JTS test (MDA-MB-231)</td><td>IC50 values ​​(nM) in the test WST SK-OV-3</td>

<td>SH-142</td><td> &lt;50</td><td> &lt;100</td><td></td>

<td>SH-143</td><td> &lt;50</td><td></td><td></td>

<td>SH-146</td><td> &lt;50</td><td></td><td></td>

<td>SH-153</td><td> &lt;10</td><td> &lt;1000</td><td></td>

<td>SH-155</td><td> &lt;10</td><td> &lt;100</td><td> &lt;100</td>

<td>SH-156</td><td> &lt;10</td><td> &lt;1000</td><td> &lt;1000</td>

<td>SH-158</td><td> &lt;10</td><td> &lt;1000</td><td> &lt;1000</td>

<td>SH-159</td><td> &lt;10</td><td> &lt;1000</td><td> &lt;1000</td>

<td>SH-164</td><td> &lt;10</td><td> &lt;10</td><td> &lt;10</td>

EP 2 019 671 B1

<td>SH-165</td><td> &lt;10</td><td> &lt;1000</td><td> &lt;1000</td>

<td>SH-166</td><td> &lt;10</td><td> &lt;1000</td><td> &lt;1000</td>

<td>SH-167</td><td> &lt;10</td><td> &lt;10</td><td> &lt;10</td>

<td>SH-172</td><td> &lt;10</td><td> &lt;1000</td><td></td>

<td>SH-173</td><td> &gt;10000</td><td> &gt;10000</td><td> &gt;10000</td>

<td>SH-175</td><td> &lt;50</td><td> &lt;100</td><td> &lt;100</td>

<td>SH-176</td><td> &lt;10</td><td> &lt;100</td><td></td>

<td>SH-177</td><td> &lt;10</td><td> &lt;100</td><td></td>

<td>SH-178</td><td> &lt;100</td><td> &lt;10000</td><td></td>

<td>SH-179</td><td> &lt;100</td><td> &lt;10000</td><td></td>

<td>SH-180</td><td> &lt;100</td><td> &lt;1000</td><td> &lt;1000</td>

<td>SH-181</td><td> &lt;100</td><td> &lt;10000</td><td></td>

<td>SH-182</td><td> &lt;100</td><td> &lt;10</td><td> &lt;10</td>

<td>SH-183</td><td> &lt;100</td><td> &lt;10000</td><td></td>

<td>SH-184</td><td> &lt;100</td><td> &lt;10</td><td> &lt;100</td>

<td>SH-185</td><td> &lt;50</td><td> &lt;10</td><td> &lt;10</td>

<td>SH-186</td><td> &lt;50</td><td> &lt;10</td><td> &lt;10</td>

<td>SH-187</td><td> &gt;10000</td><td> &gt;10000</td><td> &gt;10000</td>

<td>SH-188</td><td> &lt;100</td><td> &lt;100</td><td> &lt;100</td>

<td>SH-189</td><td> &lt;100</td><td> &lt;100</td><td> &lt;100</td>

<td>SH-190</td><td> &lt;100</td><td> &lt;10</td><td> &lt;10</td>

<td>SH-191</td><td> &lt;100</td><td> &lt;100</td><td> &lt;100</td>

<td>SH-198</td><td> &lt;100</td><td> &lt;10</td><td> &lt;10</td>

<td>SH-199</td><td> &lt;10</td><td> &lt;10</td><td> &lt;10</td>

<td>SH200</td><td> &lt;10</td><td> &lt;100</td><td> &lt;100</td>

<td>SH-201</td><td> &gt;10000</td><td> &gt;1000</td><td> &gt;1000</td>

<td>SH-202</td><td> &lt;100</td><td> &lt;10</td><td> &lt;10</td>

<td>SH-206</td><td> &lt;1000</td><td> &lt;1000</td><td> &lt;1000</td>

<td>YP-317</td><td> &lt;100</td><td> &lt;100</td><td></td>

<td>YP-343</td><td> &lt;100</td><td> &lt;100</td><td></td>

<td>YP-381</td><td> &lt;10</td><td> &lt;10</td><td> &lt;10</td>

<td>YP-383</td><td> &lt;10</td><td> &lt;10</td><td> &lt;10</td>

<td>YP-385</td><td> &lt;10</td><td> &lt;10</td><td> &lt;10</td>

<td>SM-410</td><td> &lt;100</td><td> &lt;500</td><td> &lt;500</td>

EP 2 019 671 B1