Certain chemical entities, compositions, and methods

ES 2 654 584 T3

Certain chemical entities, compositions, and methods

Description

[0001] Certain chemical entities and compositions thereof are provided that may be useful in treating cancer.

[0002] Cancer can be viewed as a breakdown in communication between tumor cells and their environment, including their normal neighboring cells. Signals, both growth-promoting and growth-inhibiting, are routinely exchanged between cells within a tissue. Cells do not normally divide in the absence of stimulating signals, and will also stop dividing in the presence of inhibitory signals. In a cancerous or neoplastic state, a cell acquires the ability to override these signals and to proliferate under conditions in which normal cells would not grow.

[0003] Bufalin is one of the predominant components of bufodienolides isolated from traditional Chinese medicine (Chan'su, toad venom), and has been found to be active against various cancer cell lines. Its anticancer activities have been reported in animal models. However, its clinical application has been limited due to its low solubility and its narrow therapeutic index.

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WO2007016656 describes methods for the treatment of eye disorders using compounds that modulate the effects of local and systemic hypoxic events.

Kamano Y. et al., Journal of Medicinal Chemistry, American Chemical Society, USA, Vol. 45, No. 25, pages 5440-5447, used QSAR analysis to identify essential structural requirements to increase the inhibitory activities of bufadienolides selected from Chinese drug Ch'an Su (and other sources) against primary liver carcinoma cell line PLC / PRF / 5 (PLC) and colchicine resistant derivative line (COL).

Kamano Y. et al., Chemical and Pharmaceutical Bulletin, Pharmaceutical Society of Japan, JP, (19880101), vol. 36, No. 1, pages 326-332, completed an evaluation of thirty-four bufadienolides and two related cardenolides against a series of rhinoviruses in vitro.

Shimada K. et al., Journal of Liquid Chromatography, New York, NY, US, (19900101), vol. 13, No. 3, pages 493-504, examined the chromatographic behavior of bufadienolides including conjugates ... by adding γ-cyclodextrin to the mobile phase in reverse phase high performance liquid chromatography.

Document CN101177445 describes new bufadienolide compound as well as the method of preparation and uses thereof. Document CN101016326 describes toad sterene compounds and their application in pharmaceutical preparation.

Liu Junshan et al., Journal of International Pharmaceutical Research, (2009), vol. 36, no. 2, pages 115-120, describes the antitumor effects of veneum bufonis and its active components.

[0004] At least one chemical entity chosen from the compounds of Formula I is provided

ES 2 654 584 T3

<img file="ES2654584T3_D0002.tif" />

and pharmaceutically acceptable salts thereof, wherein

Z is selected from ORg and NR10R11; where

Rg is selected from optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl;

R10 is selected from hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl; R11 is selected from optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl; or R10 and R11 can optionally be joined together with any intermediate atom to form an optionally substituted heterocycloalkyl ring.

[0005] Also provided is at least one chemical entity chosen from the compounds of Formula II

<img file="ES2654584T3_D0003.tif" />

and pharmaceutically acceptable salts thereof, wherein

R1 and R2 are independently selected from hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl; or R1 and R2 can optionally be joined together with any intermediate atom to form an optionally substituted heterocycloalkyl ring; for each occurrence, R3 and R4 are independently selected from hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl; or R3 and R4 can optionally be joined together with any intermediate atom to form an optionally substituted cycloalkyl ring or an optionally substituted heterocycloalkyl ring;

or R1 and an occurrence of R3 can optionally be joined together with any intermediate atom to form an optionally substituted heterocycloalkyl ring; and n is selected from 1, 2, 3, 4, 5, and 6.

[0006] Also provided is at least one chemical entity chosen from the compounds of Formula III

ES 2 654 584 T3

<img file="ES2654584T3_D0004.tif" />

and pharmaceutically acceptable salts thereof, wherein

Rs is selected from hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl; R6 is selected from optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted acyl, optionally substituted alkoxycarbonyl, and -P (= O) (OR7) (OR8), where R7 and R8 they are independently chosen from hydrogen and optionally substituted alkyl.

[0007] Also provided is a pharmaceutical composition comprising a pharmaceutically acceptable carrier and at least one chemical entity described herein.

[0008] Also provided is a packaged pharmaceutical composition comprising a pharmaceutical composition described herein and instructions for using the composition to treat a subject suffering from cancer.

[0009] Also provided is at least one chemical entity described herein for use in a method of treating cancer in a subject comprising administering to a subject in need thereof a therapeutically effective amount of at least one chemical entity described herein .

[0010] As used herein, the following words and phrases are generally intended to have the meanings set forth below, except to the extent that the context in which they are used indicates otherwise.

[0011] The following abbreviations and terms have the meanings indicated throughout:

AcOH = acetic acid

Boc = tert-butoxycarbonyl c- = cyclo

DCC = dicyclohexylcarbodiimide

DIEA = N, N-diisopropylethylamine

DMAP = 4-dimethylaminopyridine

EDC = 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide eq = equivalent (s)

Et = ethyl

EtOAc or EA = ethyl acetate

EtOH = ethanol g = gram h or hr = hour

HBTU = O- (benzotriazol-1-yl) -N, N, N ', N'-tetramethyluronium hexafluorophosphate

HOBt = hydroxybenzotriazole

HPLC = high pressure liquid chromatography i- = iso kg or Kg = kilogram

L or 1 = liter

LC / MS = LCMS = liquid chromatography-mass spectrometry LRMS = low resolution mass spectrometry m / z = mass to charge ratio

Me = methyl

MeOH = methanol

ES 2 654 584 T3 mg = milligram min = minute mL = milliliter mmol = millimole n- = normal

NaOAc = sodium acetate

PE = petroleum ether

Ph = phenyl

Prep = preparative quant. = quantitative

RP-HPLC = high pressure low pressure liquid chromatography ta or TA = room temperature s- = sec- = secondary t- = tertiary = tertiary

THF = tetrahydrofuran

TLC = thin layer chromatography

UV = ultraviolet

[0012] As used herein, when any variable appears more than once in a chemical formula, its definition in each case is independent of its definition in any other case.

[0013] As used herein, a dash (-) that is not between two letters or symbols is used to indicate a point of attachment for a substituent. For example, -CONH2 is attached through the carbon atom.

As used herein, optionally or optionally means that the event or circumstance described below may or may not occur, and that the description includes instances in which the event or circumstance occurs and instances in which it does not occur. For example, optionally substituted alkyl encompasses both alkyl and substituted alkyl as defined below. Those skilled in the art will understand, with respect to any group containing one or more substituents, that such groups are not intended to introduce any pattern of substitution or substitution that is sterically impractical, synthetically impractical, and / or inherently unstable.

[0015] As used herein, alkyl refers to straight chain and branched chain having the indicated number of carbon atoms, usually 1 to 20 carbon atoms, for example 1 to 8 carbon atoms, such as 1 to 6 carbon atoms. For example, C1-C6 alkyl encompasses straight and branched chain alkyl of 1 to 6 carbon atoms. When naming an alkyl moiety having a specific number of carbon atoms, it is intended to encompass all branched and straight chain versions having that number of carbon atoms; thus, for example, butyl is intended to include n-butyl, sec-butyl, isobutyl, and t-butyl; propyl includes n-propyl and isopropyl. Lower alkyl refers to alkyl groups having one to six carbons. Examples of alkyl groups include methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, pentyl, 2-pentyl, isopentyl, neopentyl, hexyl, 2-hexyl, 3-hexyl, 3-methylpentyl, and Similar. Alkylene is a subset of alkyl, which refers to the same residues as alkyl, but has two points of attachment. Alkylene groups will generally have 2 to 20 carbon atoms, for example 2 to 8 carbon atoms, such as 2 to 6 carbon atoms. For example, C0 alkylene indicates a covalent bond and C1 alkylene is a methylene group.

[0016] As used herein, "alkenyl" refers to a straight chain or branched unsaturated alkyl group having at least one carbon-carbon double bond derived by removal of a hydrogen molecule from adjacent alkyl carbon atoms. original. The group can be in the cis or trans configuration on the double bond. Typical alkenyl groups include, but are not limited to, ethenyl; propenyls such as prop-1-en-1-yl, prop-1-en-2-yl, prop-2-en-1-yl (allyl), prop-2-en-2-yl; butenyls such as but-1-en-1-yl, but-1-en-2-yl, 2-methyl-prop-1-en-1-yl, but-2-en-1-yl, but-2-en- 1-yl, but-2-en-2-yl, buta-1,3-dien-1-yl, buta-1,3-dien-2-yl; and the like. In certain embodiments, an alkenyl group has 2 to 20 carbon atoms and in other embodiments, 2 to 6 carbon atoms. Lower alkenyl refers to alkenyl groups having two to six carbons.

[0017] As used herein, "alkynyl" refers to a straight chain or branched unsaturated alkyl group having at least one carbon-carbon triple bond derived by the removal of two hydrogen molecules from adjacent alkyl carbon atoms. original. Typical alkynyl groups include, but are not limited to, ethynyl; propynyl such as prop-1-yn-1-yl, prop-2-y-1-yl; butynyls such as but-1-yn-1-yl, but-1-y-3-yl, but-3-y-1-yl; and the like. In certain embodiments, an alkynyl group has 2 to 20 carbon atoms and in other embodiments, 3 to 6 carbon atoms. Lower alkynyl refers to alkynyl groups that have two to six carbons.

[0018] As used herein, cycloalkyl refers to a non-aromatic carbocyclic ring, usually having 3 to 7 ring carbon atoms. The ring can be saturated or have one or more carbon-carbon double bonds. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, and cyclohexenyl, as well as bridged and caged ring groups such as norbornane.

ES 2 654 584 T3

[0019] As used herein, alkoxy refers to an alkyl group of the indicated number of carbon atoms attached through an oxygen bridge such as, for example, methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, sec-butoxy, tert. -butoxy, pentyloxy, 2-pentyloxy, isopentyloxy, neopentyloxy, hexyloxy, 2-hexyloxy, 3-hexyloxy, 3-methylpentyloxy, and the like. Alkoxy groups generally have 1 to 7 carbon atoms attached through the oxygen bridge. Lower alkoxy refers to alkoxy groups having one to six carbons.

[0020] As used herein, acyl refers to the groups HC (O) -; (alkyl) -C (O) -; (cycloalkyl) -C (O) -; (aryl) -C (O) -; (heteroaryl) -C (O) -; and (heterocycloalkyl) -C (O) -, wherein the group is attached to the parent structure through carbonyl functionality and wherein alkyl, cycloalkyl, aryl, heteroaryl, and heterocycloalkyl are as described herein. Acyl groups have the indicated number of carbon atoms, with the carbon of the keto group included in the numbered carbon atoms. For example, a C2 acyl group is an acetyl group having the formula CH3 (C = O) -.

[0021] As used herein, "formyl" refers to the group -C (O) H.

[0022] As used herein, "alkoxycarbonyl" refers to a group of the formula (alkoxy) (C = O) attached through the carbonyl carbon in which the alkoxy group has the indicated number of carbon atoms. Therefore, a C1-C6 alkoxycarbonyl group is an alkoxy group having 1 to 6 carbon atoms attached through its oxygen to a carbonyl linker.

[0023] As used herein, "azido" refers to the group -N3.

[0024] As used herein, "amino" refers to the group -NH2.

[0025] As used herein, mono- and di- (alkyl) amino refers to tertiary alkyl amino groups, where the alkyl groups are as defined above and have the indicated number of carbon atoms. The point of attachment of the alkylamino group is on the nitrogen. Examples of mono and dialkylamino groups include ethylamino, dimethylamino, and methyl-propylamino.

[0026] As used herein, aminocarbonyl refers to the group -CONR^bR^c, where

R^b is selected from H, optionally substituted C1-C6 alkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl and optionally substituted heteroaryl, optionally substituted alkoxy; Y

R^c selected from hydrogen and optionally substituted C1-C4 alkyl; or

R^b and R^c taken together with the nitrogen to which they are attached, they form an optionally substituted 5- to 7-membered nitrogen-containing heterocycloalkyl optionally including 1 or 2 additional heteroatoms selected from O, N and S on the heterocycloalkyl ring;

wherein each substituted group is independently substituted with one or more substituents independently selected from C1-C4 alkyl, aryl, heteroaryl, aryl-C1-C4 alkyl, heteroaryl-C1-C4 alkyl, halo C1-C4 alkyl, -O-C1-C4 alkyl, - OC1-C4 alkylphenyl, -C1-C4 alkyl-OH, -OC1-C4 haloalkyl, halo, -OH, -NH2, -C1-C4 alkylNH2, -N (C1-C4 alkyl) (C1-C4 alkyl), -NH (C1-C4 alkyl), -N (C1-C4 alkyl) (C1-C4 alkylphenyl), -NH (C1-C4 alkylphenyl), cyano, nitro, oxo (as a substituent for cycloalkyl, heterocycloalkyl, or heteroaryl), -CO2H, -C (O) O-C1-C4 alkyl, -CON (C1-C4 alkyl) (C1-C4 alkyl), -CONH (C1-C4 alkyl) ), -CONH2, -NHC (O) (C1-C4 alkyl), -NH (O) (phenyl), N (C1-C4 alkyl) C (O) (C1-C4 alkyl), -N (C1-alkyl) C4) C (O) (phenyl), -C (O) C1-C4 alkyl, -C (O) C1-C4 alkylphenyl, C (O) C1-C4 haloalkyl, -OC (O) C1-C4 alkyl, - SO2 (C1-C4 alkyl), -SO2 (phenyl), -SO2 (C1-C4 haloalkyl), -SO2NH2, SO2NH (C1-C4 alkyl), -SO2NH (phenyl), -NHSP2 (C1-C4 alkyl), -NHSO2 (phenyl), and -NHSO2 (C1-C4 haloalkyl).

[0027] As used herein, aryl refers to: 6-membered carbocyclic aromatic rings, eg, benzene; bicyclic ring systems in which at least one ring is carbocyclic and aromatic, eg, naphthalene, indane, and tetralin; and tricyclic ring systems in which at least one ring is carbocyclic and aromatic, eg, fluorene.

[0028] For example, aryl includes 6-membered carbocyclic aromatic rings fused to a 5- to 7-membered heterocycloalkyl ring containing 1 or more heteroatoms selected from N, O and S. For said fused bicyclic ring systems in which only one of the rings are a carbocyclic aromatic ring, the point of attachment can be the carbocyclic aromatic ring or the heterocycloalkyl ring. The divalent radicals formed from substituted benzene derivatives and having the free valences at the ring atoms are referred to as substituted phenylene radicals. Bivalent radicals derived from univalent polycyclic hydrocarbon radicals whose names end in -yl by removing a hydrogen atom from the carbon atom with the free valence are named by adding -idene to the name of the corresponding univalent radical, for example, a naphthyl group. with two points of attachment is called naphthylidene. Aryl, however, does not encompass or overlap in any way with heteroaryl, defined separately below. Therefore, if one or more carbocyclic aromatic rings are fused to an aromatic heterocycloalkyl ring, the resulting ring system is heteroaryl, not aryl, as defined herein.

[0029] As used herein, "aryloxy" refers to the group -O-aryl.

ES 2 654 584 T3

[0030] As used herein, "aralkyl" refers to the group -alkyl-aryl.

[0031] As used herein, carbamimidoyl refers to the group -C (= NH) -NH2.

[0032] As used herein, substituted carbamimidoyl refers to the group -C (= NR^and) -NR^FR^g where R^and selected from hydrogen, cyano, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, and optionally substituted heterocycloalkyl; and R^F and R^g are independently selected from hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, and optionally substituted heterocycloalkyl, provided that at least one of R^and, R^F and R^g is not hydrogen and where substituted alkyl, cycloalkyl, aryl, heterocycloalkyl and heteroaryl refer respectively to alkyl, cycloalkyl, aryl, heterocycloalkyl and heteroaryl where one or more (such as up to 5, for example, up to 3) hydrogen atoms are replaced by a substituent independently chosen from

-R^to, -OR^b, optionally substituted amino (including -NR^cCOR^b, -NR^cCÜ2R^to, -NR^cCONR^bR^c, -NR^bC (NR^c) NR^bR^c, NR^bC (NCN) NR^bR^c and -NR^cSO2R^to), halo, cyano, nitro, oxo (as a substituent for cycloalkyl, heterocycloalkyl, and heteroaryl), optionally substituted acyl (such as -COR^b), optionally substituted alkoxycarbonyl (such as CO2R^b), aminocarbonyl (such as -CONR^bR^c), -OCOR^b, -OCO2R^to, -OCONR^bR^c, -OP (O) (OR^b) OR^c sulfanyl (as SR^b), sulfinyl (as -SOR^to) and sulfonyl (as -SO2R^to and -SO2NR^bR^c), where R^to selected from optionally substituted C1-C6 alkyl, optionally substituted aryl, and optionally substituted heteroaryl;

R^b selected from H, optionally substituted C1-C6 alkyl, optionally substituted aryl, and optionally substituted heteroaryl; Y

R^c selected from hydrogen and optionally substituted C1-C4 alkyl; or

R^b and R^c, and the nitrogen to which they are attached, form an optionally substituted heterocycloalkyl group; and where each optionally substituted group is unsubstituted or is independently substituted with one or more, such as one, two, or three, substituents independently selected from C1-C4 alkyl, aryl, heteroaryl, aryloC1-C4 alkyl, heteroaryl-C1-C4 alkyl. -, C1-C4 haloalkyl, -OC1-C4 alkyl, -OC1-C4 alkylphenyl, -C1-C4 alkyl-OH, OC1-C4 haloalkyl, halo, -OH, -NH2, C1-C4 alkyl-NH2, -N ( C1-C4 alkyl) (C1-C4 alkyl), -NH (C1-C4 alkyl), N (C1-C4 alkyl) (C1-C4 alkylphenyl), -NH (C1-C4 alkylphenyl), cyano, nitro, oxo (as a cycloalkyl, heterocycloalkyl or heteroaryl substituent), -CO2H-C (O) O-C1 alkyl -C4, -CON (C1-C4 alkyl) (C1-C4 alkyl), -CONH (C1-C4) alkyl), -CONH2, -NHC (O) (C1-C4 alkyl), -NHC (O) (phenyl ), -N (C1-C4 alkyl) C (O) (C1-C4 alkyl), -N (C1 -C4 alkyl) C (O) (phenyl), -C (O) C1-C4 alkyl, -C ( O) C1-C4 phenyl, -C (O) C1-C4 haloalkyl, -OC (O) C1-C4 alkyl, -SO2 (C1-C4 alkyl), -SO2 (phenyl), -SO2 (C1-C4 haloalkyl), -SO2NH2, -SO2NH (C1-C4 alkyl), -SO2 NH (phenyl), -NHSO2 (C1-C4 alkyl), -NHSO2 (phenyl) and -NHSO2 (C1-C4 haloalkyl) ).

[0033] As used herein, halo refers to fluorine, chlorine, bromine, and iodine, and the term halogen includes fluorine, chlorine, bromine, and iodine.

[0034] As used herein, "haloalkyl" refers to alkyl as defined above having the specified number of carbon atoms, substituted with 1 or more halogen atoms, up to the maximum allowed number of halogen atoms. Examples of haloalkyl include, but are not limited to, trifluoromethyl, difluoromethyl, 2-fluoroethyl, and pentafluoroethyl.

[0035] As used herein, heteroaryl refers to:

5- to 7-membered aromatic monocyclic rings containing one or more, eg, 1 to 4, or in certain embodiments, 1 to 3, heteroatoms selected from N, O, and S, with the remaining ring atoms being carbon; Bicyclic heterocycloalkyl rings containing one or more, for example 1 to 4, or in certain embodiments, 1 to 3, heteroatoms selected from N, O and S, the remaining ring atoms being carbon and where at least one heteroatom is present on an aromatic ring; and tricyclic heterocycloalkyl rings containing one or more, for example, 1 to 5, or in certain embodiments, 1 to 4, heteroatoms selected from N, O, and S, the remaining ring atoms being carbon and in which at least a heteroatom is present on an aromatic ring.

[0036] For example, heteroaryl includes a 5 to 7 membered heterocycloalkyl ring fused to a 5 to 7 membered cycloalkyl or heterocycloalkyl ring. For such fused bicyclic heteroaryl ring systems in which only one of the rings contains one or more heteroatoms, the point of attachment can be on either ring. When the total number of S and O atoms in the heteroaryl group exceeds 1, those heteroatoms are not adjacent to each other. In certain embodiments, the total number of S and O atoms in the heteroaryl group is not more than 2. In certain embodiments, the total number of S and O atoms in the aromatic heterocycle is not more than 1. Examples of heteroaryl groups include, but are not limited to, (as numbered from the assigned priority 1 binding position), 2-pyridyl, 3-pyridyl, 4-pyridyl, 2,3-pyrazinyl, 3,4-pyrazinyl, 2,4-pyrimidinyl, 3,5-pyrimidinyl, 2,3-pyrazolinyl, 2,4-imidazolinyl, isoxazolinyl, oxazolinyl, thiazolinyl, thiadiazolinyl, tetrazolyl, thienyl, benzothiophenyl, furanyl, benzofuranyl, benzoimidazolinyl, indolinyl, pyridazinyl, quinoline pyrazolyl and 5,6, 7.8tetrahydroisoquinolinyl. The bivalent radicals derived from univalent heteroaryl radicals whose names

ES 2 654 584 T3 terminate in -yl by removing a hydrogen atom from the atom with the free valence are named by adding -idene to the name of the corresponding univalent radical, for example, a pyridyl group with two points of attachment is a pyridylidene. Heteroaryl does not encompass or overlap with aryl, cycloalkyl, or heterocycloalkyl, as defined herein.

[0037] Substituted heteroaryl also includes ring systems substituted with one or more oxide (-O-) substituents, such as pyridinyl N-oxides.

[0038] As used herein, heterocycloalkyl refers to a single non-aromatic ring, usually with 3 to 7 ring atoms, containing at least 2 carbon atoms in addition to 1-3 heteroatoms independently chosen from oxygen, sulfur and nitrogen, as well as combinations comprising at least one of the above heteroatoms. The ring can be saturated or have one or more carbon-carbon double bonds. Suitable heterocycloalkyl groups include, for example (as numbered from the assigned priority linking position 1), 2-pyrrolidinyl, 2,4-imidazolidinyl, 2,3-pyrazolidinyl, 2-piperidyl, 3-piperidyl, 4-piperidyl and 2,5piperizinyl. Morpholinyl groups are also contemplated, including 2-morpholinyl and 3-morpholinyl (numbered with priority given to oxygen 1). Substituted heterocycloalkyl also includes ring systems substituted with one or more oxo (= O) or oxide (-O-) substituents, such as piperidinyl N-oxide, morpholinyl N-oxide, 1-oxo-1-thiomorpholinyl and 1,1- dioxo-1-thiomorpholinyl.

[0039] Heterocycloalkyl also includes bicyclic ring systems wherein a non-aromatic ring, usually with 3 to 7 ring atoms, contains at least 2 carbon atoms in addition to 1-3 heteroatoms independently chosen from oxygen, sulfur and nitrogen, thus as combinations comprising at least one of the above heteroatoms; and the other ring, usually with 3 to 7 ring atoms, optionally contains 1-3 heteroatoms independently chosen from oxygen, sulfur and nitrogen and is non-aromatic.

[0040] As used herein, sulfanyl refers to the groups: -S- (optionally substituted (Ci-Ce) alkyl), -S- (optionally substituted aryl), -S- (optionally substituted heteroaryl) and -S- (optionally substituted heterocycloalkyl). Thus, sulfanyl includes the C1-C6 alkylsulfanyl group.

[0041] As used herein, sulfinyl refers to the groups: -S (O) - (optionally substituted (C1-C6) alkyl), -S (O) -optionally substituted aryl), -S (O) -optionally substituted heteroaryl), -S (O) - (optionally substituted heterocycloalkyl); and -S (O) - (optionally substituted amino).

[0042] As used herein, "sulfonyl" refers to the groups: -S (O2) - (optionally substituted (Ci-Ce) alkyl), -S (O2) -optionally substituted aryl), -S (O2) -optionally substituted heteroaryl), -S (O2) (optionally substituted heterocycloalkyl) and -S (O2) - (optionally substituted amino).

[0043] As used herein, "substituted" refers to that one or more hydrogens on the designated atom or group is replaced with a selection from the indicated group, provided that the normal valence of the designated atom is not exceeded. When a substituent is oxo (i.e. = O), 2 hydrogens on the atom are replaced. Combinations of substituents and / or variables are permissible only if such combinations result in stable compounds or useful synthetic intermediates. A stable compound or stable structure is understood to imply a compound that is sufficiently robust to survive isolation from a reaction mixture, and further formulation as an agent that has at least practical utility. Unless otherwise specified, substituents are named in the core structure. For example, it should be understood that when (cycloalkyl) alkyl is mentioned as a possible substituent, the point of attachment of this substituent to the core structure is in the alkyl part.

[0044] As used herein, the terms alkyl, cycloalkyl, aryl, heterocycloalkyl, and substituted heteroaryl, unless expressly defined otherwise, refer respectively to alkyl, cycloalkyl, aryl, heterocycloalkyl, and heteroaryl in which one or more (such as up to 5, for example, up to 3) hydrogen atoms are replaced by a substituent independently chosen from

-R^to, -OR^b, optionally substituted amino (including -NR^cCOR^b, -NR^cCO2R^to, -NR^cCONR^bR^c, -NR^bC (NR^c) NR^bR^c, NR^bC (NCN) NR^bR^c and -NR^cSO2R^to), halo, cyano, azido, nitro, oxo (as a substituent for cycloalkyl or heterocycloalkyl), optionally substituted acyl (such as -COR^b), optionally substituted alkoxycarbonyl (such as -CO2R^b), aminocarbonyl (such as -CONR^bR^c), -OCOR^b, -OCO2R^to, -OCONR^bR^c, -OP (O) (OR^b) OR^c, sulfanyl (as SR^b), sulfinyl (as -SOR^to) and sulfonyl (as -SO2Ra and -SO2NR^bR^c), where

R^to selected from optionally substituted C1-C6 alkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, and optionally substituted heteroaryl; R^b selected from hydrogen, optionally substituted C1-C6 alkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl; Y

R^c selected from hydrogen and optionally substituted C1-C4 alkyl; or

R^b and R^c, and the nitrogen to which they are attached, form an optionally substituted heterocycloalkyl group; and where each optionally substituted group is unsubstituted or is independently substituted with one or more, such as one, two, or three, substituents independently selected from C1-C4 alkyl, aryl, heteroaryl,

ES 2 654 584 T3 aryl-C1-C4 alkyl, heteroaryl-Ci-C4 alkyl-, C1-C4 haloalkyl, -OC1-C4 alkyl, -OC1-C4 alkylphenyl, -C1-C4 alkylOH, -OC1-C4 haloalkyl, halo , -OH, -NH2, C1-C4 alkyl-NH2, -N (C1-C4 alkyl) (C1-C4 alkyl), -NH (C1-C4 alkyl), N (C1-C4 alkyl) (C1-C4 alkylphenyl ), -NH (C1-C4 alkylphenyl), cyano, nitro, oxo (as a substituent for cycloalkyl or heterocycloalkyl), -CO2H, -C (O) OC1-C4 alkyl, -CON (C1-C4 alkyl) (C1-alkyl) C4), -CONH (C1-C4 alkyl), -CONH2, -NHC (O) (C1-C4 alkyl), -NHC (O) (phenyl), -N (C1-C4 alkyl) C (O) (C1-C4 alkyl), -N (C1-C4) alkyl) C (O) (phenyl), C (O) C1-C4 alkyl, -C (O) C1-C4 alkylphenyl, -C (O) C1-C4 haloalkyl, -OC (O) C1-C4 alkyl, -SO2 (alkyl C1-C4), -SO2 (phenyl), -SO2 (C1-C4 haloalkyl), -SO2NH2, -SO2NH (C1-C4 alkyl), -SO2NH (phenyl), -NHSO2 (C1-C4 alkyl), - NHSO2 ( phenyl) and -NHSO2 (C1-C4 haloalkyl).

[0045] As used herein, substituted acyl refers to the groups (substituted alkyl) -C (O) -; (substituted cycloalkyl) -C (O) -; (substituted aryl) -C (O) -; (substituted heteroaryl) -C (O) -; and (substituted heterocycloalkyl) C (O) -, wherein the group is attached to the parent structure through carbonyl functionality and wherein substituted alkyl, cycloalkyl, aryl, heteroaryl, and heterocycloalkyl refer to alkyl, cycloalkyl, aryl, respectively. , heteroaryl and heterocycloalkyl in which one or more (such as up to 5, for example, up to 3) hydrogen atoms are replaced by a substituent independently chosen from -R^to, -OR^b, optionally substituted amino (including -NR^cCOR^b, -NR^cCO2R^to, - NRcCONR^bR^c, -NR^bC (NR^c) NR^bR^c, -NR^bC (NCN) NR^bR^c and -NR^cSO12R^to), halo, cyano, nitro, oxo (as a substituent for cycloalkyl or heterocycloalkyl), optionally substituted acyl (such as -COR^b), optionally substituted alkoxycarbonyl (as -CO2R^b), aminocarbonyl (as CONR^bR^c), -OCOR^b, -OCO2R^to, -OCONR^bR^c, -OP (O) (OR^b) OR^c, sulfanyl (as SR^b), sulfinyl (as -SOR^to), and sulfonyl (as -SO2R^to and -SO2NR^bR^c), where R^to selected from optionally substituted C1-C6 alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, and optionally substituted heteroaryl;

R^b selected from H, optionally substituted C1-C6 alkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl; and R^c selected from hydrogen and optionally substituted C1-C4 alkyl; or

R^b and R^c, and the nitrogen to which they are attached, form an optionally substituted heterocycloalkyl group; and wherein each optionally substituted group is unsubstituted or is independently substituted with one or more, such as one, two, or three, substituents independently selected from C1-C4 alkyl, aryl, heteroaryl, aryloC1-C4 alkyl, heteroaryl-C1-C4 alkyl. -, C1-C4 haloalkyl, -OC1-C4 alkyl, -OC1-C4 alkylphenyl, -C1-C4 alkyl-OH, OC1-C4 haloalkyl, halo, -OH, -NH2, C1-C4 alkyl-NH2, -N ( C1-C4 alkyl) (C1-C4 alkyl), -NH (C1-C4 alkyl), N (C1-C4 alkyl) (C1-C4 alkylphenyl), -NH (C1-C4 alkylphenyl), cyano, nitro, oxo (as cycloalkyl or heterocycloalkyl substituent), -CO2H-C (O) O-C1-C4 alkyl , -CON (C1-C4 alkyl) (C1-C4 alkyl), -CONH (C1-C4 alkyl), -CONH2, NHC (O) (C1-C4 alkyl), -NHC (O) (phenyl), -N (C1-C4 alkyl) C (O) (C1-C4 alkyl), -N (C1-C4 alkyl) C (O) (phenyl), C (O) C1-C4 alkyl, -C (O) C1-C alkylphenyl C4, -C (O) C1-C4 haloalkyl, -OC (O) C1-C4 alkyl, -SO2 (C1-C4 alkyl), -SO2 (phenyl), -SO2 (C1-C4 haloalkyl), -SO2NH2-SO2NH (C1-C4 alkyl), -SO2NH (phenyl), -NHSO2 (C1-C4 alkyl), -NHSO2 (phenyl) and -NHSO2 (C1-C4 haloalkyl).

[0046] As used herein, substituted alkoxy refers to alkoxy in which the alkyl constituent is substituted (ie, -O- (substituted alkyl)) in which substituted alkyl refers to alkyl in which one or plus (such as up to 5, for example, up to 3) hydrogen atoms are replaced by a substituent independently chosen from

-R^to, -OR^b, optionally substituted amino (including -NR^cCOR^b, -NR^cCO2R^to, -NR^cCONR^bR^c, -NR^bC (NR^C) NR^bR^c, NR^bC (NCN) NR^bR^c, and -NR^cSO2R^to), halo, cyano, nitro, oxo (as a substituent for cycloalkyl or heterocycloalkyl), optionally acyl (such as -COR^b), optionally substituted alkoxycarbonyl (such as -CO2R^b), aminocarbonyl (such as -CONR^bR^c), -OCOR^b, -OCO2R^to, -OCONR^bR^c, -OP (O) substituted (OR^b) OR^c, sulfanyl (for example, SR^b), sulfinyl (for example, -SOR^to) and sulfonyl (such as -SO2R^to and -SO2NR^bR^c), where R^to selected from optionally substituted C1-C6 alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, and optionally substituted heteroaryl;

R^b selected from H, optionally substituted C1-C6 alkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl; and R^c chosen from hydrogen and optionally substituted C1-C4 alkyl; or

R^b and R^c, and the nitrogen to which they are attached, form an optionally substituted heterocycloalkyl group; and wherein each optionally substituted group is independently substituted or substituted with one or more, such as one, two, or three, substituents independently chosen from C1-C4 alkyl, aryl, heteroaryl, aryl-C1-C4 alkyl, heteroaryl-C1- C4 alkyl, C1-C4 haloalkyl, -OC1-C4 alkyl, -OC1-C4 alkylphenyl, -C1-C4 alkyl-OH, -OC1-C4 haloalkyl, halo, -Oh, -NH2, -C1-C4 alkyl alkyl- NH2, -N (C1-C4 alkyl) (C1-C4 alkyl), -NH (C1-C4 alkyl), N (C1-C4 alkyl) (C1-C4 alkylphenyl), -NH (C1-C4 alkylphenyl), cyano, nitro, oxo (as a substituent for cycloalkyl or heterocycloalkyl), -CO2H, -C (O) OC1-C4 alkyl , -CON (C1-C4 alkyl) (C1-C4 alkyl), -CONH (C1-C4 alkyl), -CONH2, -NHC (O) (C1-C4 alkyl), -NHC (O) (phenyl), -N ( C1-C4 alkyl) C (O) (C1-C4 alkyl), -N (C1-C4 alkyl) C (O) (phenyl), -C (O) C1-C4 alkyl, -C (O) C1-alkylphenyl C4, -C (O) C1-C4 haloalkyl, -OC (O) C1-C4 alkyl, -SO2 (C1-C4 alkyl), -SO2 (phenyl), -SO2 (C1-C4 haloalkyl), -SO2NH2 -SO2NH (C1-C4 alkyl), -SO2NH (phenyl), NHSO2 (C1-C4 alkyl), -NHSO2 (phenyl), and -NHSO2 (C1-C4 haloalkyl).

In some embodiments, a substituted alkoxy group is polyalkoxy or -O- (optionally substituted alkylene) (optionally substituted alkoxy), and includes groups such as -OCH2CH2OCH3, and glycol ethers residues such as polyethylene glycol, and - Or (CH2CH2OXCH3, where x is an integer from 2-20, such as 2-10, and for example, 29

ES 2 654 584 T3

5. Another substituted alkoxy group is hydroxylalkoxy or -OCH2 (CH2) yOH, where y is an integer from 1-10, such as 1-4.

[0048] As used herein, substituted alkoxycarbonyl refers to the group (substituted alkyl) -OC (O) - in which the group is attached to the parent structure through carbonyl functionality and in which substituted refers to alkyl in which one or more (such as up to 5, for example, up to 3) hydrogen atoms are replaced by a substituent independently chosen from -R^to, -OR^b, optionally substituted amino (including -NR^cCOR^b, -NR^cCO2R^to, -NR^cCONR^bR^c, -NR^bC (NR^C) NR^bR^c, - NRBC (NCN) NR^bR^c, and -NR^cSO2R^to), halo, cyano, nitro, oxo (as a substituent for cycloalkyl or heterocycloalkyl), optionally acyl (such as -COR^b), substituted alkoxycarbonyl (such as -CO2Rb), substituted aminocarbonyl (such as -CONR^bR^c), OCOR^b, -OCO2R^to, -OCONR^bR^c, -OC (O) (OR^b) OR^c, sulfanyl (for example, SR^B), sulfinyl (for example, -SOR^to) and sulfonyl (such as -SO2Ra and -SO2NR^bR^c), where R^to selected from optionally substituted C1-C6 alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, and optionally substituted heteroaryl;

R^b selected from H, optionally substituted C1-C6 alkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl; and R^c chosen from hydrogen and optionally substituted C1-C4 alkyl; or

R^b and R^c, and the nitrogen to which they are attached, form an optionally substituted heterocycloalkyl group; and wherein each optionally substituted group is independently substituted or substituted with one or more, such as one, two, or three, substituents independently chosen from C1-C4 alkyl, aryl, heteroaryl, aryl-C1-C4 alkyl, heteroaryl-C1- C4 alkyl, C1-C4 haloalkyl, -OC1-C4 alkyl, -OC1-C4 alkylphenyl, -C1-C4 alkyl-OH, OC1-C4 haloalkyl, halo, -OH, -NH2, -C1-C4 alkyl-NH2, -N (C1-C4 alkyl) (C1-C4 alkyl), -NH (C1-C4 alkyl), -N (C1-C4 alkyl) (C1-C4 alkylphenyl), -NH (C1-C4 alkylphenyl), cyano, nitro, oxo (as a substituent for cycloalkyl or heterocycloalkyl), -CO2H, -C (O) OC1-C4 alkyl , -CON (C1-C4 alkyl) (C1-C4 alkyl), -CONH (C1-C4 alkyl), -CONH2, -NHC (O) (C1-C4 alkyl), -NHC (O) (phenyl), - N (C1-C4 alkyl) C (O) (C1-C4 alkyl), -N (C-1-C4 alkyl) C (O) (phenyl), -C (O C1-C4 alkyl, -C (O) ) C1-C4 alkyl phenyl, -C (O) C1-C4 haloalkyl, -OC (O) C1-C4 alkyl, -SO2 (C1-C4 alkyl), -SO2 (phenyl), -SO2 (C1-C4 haloalkyl), -SO2NH2 -SO2NH (C1-C4 alkyl), -SO2NH (phenyl), -NHSO2 (C1-C4 alkyl), -NHSO2 (phenyl), and -NHSO2 (C1-C4 haloalkyl) ).

[0049] As used herein, "substituted amino" refers to the group -NHR^d or -NR^dR^and in which R^d is selected from hydroxy, formyl, optionally substituted alkoxy, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted acyl, optionally substituted carbamimidoyl, aminocarbonyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocycloalkyl, optionally substituted alkoxycarbonyl, sulfinyl, and sulfonyl and in which R^and is selected from optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, and optionally substituted heterocycloalkyl, and wherein substituted alkyl, cycloalkyl, aryl, heterocycloalkyl, and heteroaryl refer respectively to alkyl, cycloalkyl, aryl, heterocycloalkyl and heteroaryl in which one or more (such as up to 5, for example, up to 3) hydrogen atoms are replaced by an independently chosen substituent from

-R^to, -OR^b, optionally substituted amino (including -NR^cCOR^b, -NR^cCO2R^to, -NR^cCONR^bR^c, -NR^bC (NR^c) NR^bR^c, NR^bC (NCN) NR^bR^c, and -NR^cSO2R^to), halo, cyano, nitro, oxo (as a substituent for cycloalkyl or heterocycloalkyl), optionally acyl (such as -COR^b), optionally substituted alkoxycarbonyl (such as -CO2R^b), aminocarbonyl (such as -CONR^bR^c), -OCOR^b, -OCO2R^to, -OCONR^bR^c, -OP (O) substituted (OR^B) OR^c, sulfanyl (for example, SR^B), sulfinyl (for example, -SOR^to) and sulfonyl (such as -SO2R^to and -SO2NR^bR^c), where R^to selected from optionally substituted C1-C6 alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, and optionally substituted heteroaryl;

R^b selected from H, optionally substituted C1-C6 alkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl; and R^c chosen from hydrogen and optionally substituted C1-C4 alkyl; or

R^b and R^c, and the nitrogen to which they are attached, form an optionally substituted heterocycloalkyl group; and wherein each optionally substituted group is independently substituted or substituted with one or more, such as one, two, or three, substituents independently chosen from C1-C4 alkyl, aryl, heteroaryl, aryl-C1-C4 alkyl, heteroaryl-C1- C4 alkyl, C1-C4 haloalkyl, -OC1-C4 alkyl, -OC1-C4 alkylphenyl, C1-C4 alkyl-OH, -OC1-C4 haloalkyl, halo, -OH, -NH2, -C1-C4 alkyl-NH2, -N (C1-C4 alkyl) (C1-C4 alkyl), -NH (C1-C4 alkyl), N (C1-C4 alkyl) (C1-C4 alkylphenyl), -NH (C1-C4 alkylphenyl), cyano, nitro, oxo (as a substituent for cycloalkyl or heterocycloalkyl), -CO2H, -C (O) OC1-C4 alkyl , -CON (C1-C4 alkyl) (C1-C4 alkyl), -CONH (C1-C4 alkyl), -CONH2, -NHC (O) (C1-C4 alkyl), -NHC (O) (phenyl), -N ( C1-C4 alkyl) C (O) (C1-C4 alkyl), -N (C1-C4 alkyl) C (O) (phenyl), -C (O) C1-C4 alkyl, -C (O) C1-C4 alkyl phenyl , -C (O) C1-C4 haloalkyl, -OC (O) C1-C4 alkyl, SO2 (C1-C4 alkyl), -SO2 (phenyl), -SO2 (C1-C4 haloalkyl), -SO2NH2, -SO2NH (C1-C4 alkyl), -SO2NH (phenyl), NHSO2 (C1-C4 alkyl), -NHSO2 (phenyl), and -NHSO2 (C1-C4 haloalkyl) ; and where optionally substituted acyl, optionally substituted alkoxycarbonyl, sulfinyl, and sulfonyl are as defined herein.

[0050] The term substituted amino also refers to the N-oxides of the -NHR groups^d, and NRdRd each as described above. N-oxides can be prepared by treating the corresponding amino group with, for example, hydrogen peroxide or m-chloroperoxybenzoic acid. The person skilled in the art is

ES 2 654 584 T3 familiar with the reaction conditions to carry out the N-oxidation.

[0051] The compounds described herein include, but are not limited to, their optical isomers, racemates, and other mixtures thereof. In those situations, the individual enantiomers or diastereomers, ie, optically active forms, can be obtained by asymmetric synthesis or by resolution of the racemates. Resolution of the racemates can be achieved, for example, by conventional methods such as crystallization in the presence of a resolving agent, or chromatography, using, for example, a chiral high pressure liquid chromatography (HPLC) column. In addition, the compounds include Z and E forms (or cis and trans forms) of the compounds with carbon-carbon double bonds. When compounds described herein exist in various tautomeric forms, the term "compound" is intended to include all tautomeric forms of the compound.

The compounds of Formula I-III also include crystalline and amorphous forms of the compounds, including, for example, polymorphs, pseudopolymorphs, solvates (including hydrates), unsolvated polymorphs (including anhydrates), conformational polymorphs, and amorphous forms of the compounds, as well as mixtures thereof. Crystalline, polymorphic, and new forms can be used interchangeably herein, and are understood to include crystalline and amorphous forms of the compound, including, for example, polymorphs, pseudopolymorphs, solvates (including hydrates), unsolvated polymorphs (including anhydrates). , conformational polymorphs, and amorphous forms, as well as mixtures thereof, unless a particular or amorphous crystalline is known. Similarly, pharmaceutically acceptable salts of compounds of Formula IIII also include crystalline and amorphous forms of the compounds, including, for example, polymorphs, pseudopolymorphs, solvates (including hydrates), unsolvated polymorphs (including anhydrates), conformational polymorphs, and forms amorphous pharmaceutically acceptable salts, as well as their mixtures.

[0053] A solvate is formed by the interaction of a solvent and a compound. The term compound is intended to include solvates of the compounds. Similarly, pharmaceutically acceptable salts include pharmaceutically acceptable salt solvates. Suitable solvates are pharmaceutically acceptable solvates, such as hydrates, including monohydrates and hemihydrates.

The compounds of Formula I-III also include other pharmaceutically acceptable forms of the recited compounds, including chelates, non-covalent complexes, and mixtures thereof.

[0055] A chelate is formed by the coordination of a compound to a metal ion at two (or more) points. The term compound is intended to include the chelates of compounds. Similarly, pharmaceutically acceptable salts include chelates of pharmaceutically acceptable salts.

[0056] A non-covalent complex is formed by the interaction of a compound and another molecule in which a covalent bond is not formed between the compound and the molecule. For example, complexation can occur through van der Waals interactions, hydrogen bonding, and electrostatic interactions (also called ionic bonding). Such non-covalent complexes are included in the term compound '. Similarly, 'pharmaceutically acceptable salts' includes 'non-covalent complexes' of pharmaceutically acceptable salts.

The term hydrogen bonding refers to a form of association between an electronegative atom (also known as a hydrogen bond acceptor) and a hydrogen atom bonded to a second, relatively electronegative atom (also known as a donor of hydrogen bond). Suitable hydrogen bond donor and acceptors are well known in medicinal chemistry.

[0058] Hydrogen bond acceptor refers to a group comprising an oxygen or nitrogen atom, such as an oxygen or nitrogen that is sp²-hybridized, an ether oxygen, or the oxygen of a sulfoxide or N-oxide.

The term "hydrogen bond donor" refers to a heteroaromatic oxygen, nitrogen, or carbon bearing a hydrogen group containing a ring nitrogen or a heteroaryl group containing a ring nitrogen.

[0060] The compounds described herein can be used in different enriched isotopic forms, for example enriched in the content of ²H, ³H, ¹¹C, ¹³C and / or ¹⁴C. In a particular embodiment, the compound deuters in at least one position. Such deuterated forms can be made by the procedure described in United States Patent No.^you 5,846,514 and 6,334,997. As described in US Patent No.^you 5,846,514 and 6,334,997, deuteration can improve the efficacy and increase the duration of action of drugs.

[0061] Substituted deuterium compounds can be synthesized using various methods such as described in: Dean, Dennis C .; Editor. Recent advances in the synthesis and applications of radiolabeled compounds for drug discovery and development. [In :. Curr, Pharm. Des., 2000; 6 (10)] 2000, 110 pp; George W .; Varma, Rajender S. The Synthesis of Radiolabeled Compounds via Organometallic Intermediates, Tetrahedron, 1989, 45 (21), 6601-21; and Evans, E. Anthony. Synthesis of radiolabeled compound, J. Radioanal. Chem., 1981, 64 (1-2), 9-32.

ES 2 654 584 T3

[0062] Pharmaceutically acceptable salts Include, but are not limited to, salts with Inorganic acids, such as hydrochlorate, phosphate, diphosphate, hydrobromide, sulfate, sulphinate, nitrate, and as salts; as well as salts with an organic acid, such as malate, maleate, fumarate, tartrate, succinate, citrate, acetate, lactate, methanesulfonate, ptoluenesulfonate, 2-hydroxyethylsulfonate, benzoate, salicylate, stearate, and alkanoate, such as acetate, HOOC- ( CH2) n-COOH where n is 0-4, and like salts. Similarly, pharmaceutically acceptable cations include, but are not limited to, sodium, potassium, calcium, aluminum, lithium, and ammonium.

Furthermore, if the compounds described herein are obtained as an acid addition salt, the free base can be obtained by basifying a solution of the acid salt. Conversely, if the product is a free base, an addition salt, particularly a pharmaceutically acceptable addition salt, can be produced by dissolving the free base in a suitable organic solvent and treating the solution with an acid, according to conventional procedures. to prepare acid addition salts of base compounds. Those skilled in the art will recognize various synthetic methodologies that can be used to prepare non-toxic pharmaceutically acceptable addition salts.

[0064] As used herein, the terms group, radical or fragment are synonymous and are intended to indicate groups or fragments of functional molecules connectable to a bond or other fragments of molecules.

[0065] As used herein, modulation refers to a change in activity as a direct or indirect response to the presence of a chemical entity as described herein, relative to activity in the absence of the entity. chemistry. The change can be an increase in activity or a decrease in activity, and it can be due to the direct interaction of the compound with that of a target or due to the interaction of the compound with one or more other factors that in turn affect to the activity of the target. For example, the presence of the chemical entity can, for example, increase or decrease the target activity by binding directly to the target, causing (directly or indirectly) another factor to increase or decrease the target activity, or increasing (directly or indirectly) ) or decreasing the amount of target present in the cell or organism.

[0066] As used herein, active agent is used to denote a chemical entity that has biological activity. In certain embodiments, an active agent is a compound that has pharmaceutical utility. For example, an active agent can be an anticancer therapeutic agent.

[0067] As used herein, significant refers to any detectable change that is statistically significant in a standard parametric test of statistical significance such as Student's t-test, where p &lt;0.05.

[0068] As used herein, a pharmaceutically acceptable component is one that is suitable for use with humans and / or animals without undue adverse side effects (such as toxicity, irritation, and allergic response) commensurate with a benefit / risk ratio. reasonable.

[0069] As used herein, therapeutically effective amount of a chemical entity described herein refers to an amount effective, when administered to a human or non-human subject, to provide a therapeutic benefit such as enhancement of the symptoms, slowing disease progression, or disease prevention.

Treating or treatment encompasses the administration of at least one compound of Formula I-III, or a pharmaceutically acceptable salt thereof, to a mammalian subject, particularly a human subject, in need of such administration and includes (i) arresting the development of clinical symptoms of the disease, such as cancer, (ii) cause a regression of clinical symptoms of the disease, such as cancer, and / or (iii) prophylactic treatment to prevent the onset of diseases, like cancer.

[0071] As used herein, "cancer" refers to all types of cancer or neoplasm or malignant tumors found in mammals, including carcinomas and sarcomas. Examples of cancer are cancer of the brain, breast, cervix, colon, head and neck, kidney, lung, non-small cell lung, melanoma, mesothelioma, ovary, sarcoma, stomach, uterus, and medulloblastoma.

[0072] As used herein, "subject" refers to a mammal that has been or will be the object of treatment, observation, or experiment. The chemical entities described herein may be for use in human therapy and veterinary applications. In some embodiments, the subject is a human.

The term "mammal" is intended to have its standard meaning, and encompasses humans, dogs, cats, sheep, and cows, for example.

[0074] Provided is at least one chemical entity selected from compounds of Formula I

ES 2 654 584 T3

<img file="ES2654584T3_D0005.tif" />

and pharmaceutically acceptable salts thereof, wherein

Z is selected from ORg and NR10R11; where

Rg is selected from optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl; R10 is selected from hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl; R11 is selected from optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl; or R10 and R11 may be optionally linked together with any Intermediate atoms to form an optionally substituted heterocycloalkyl ring.

[0075] In some embodiments, Z is ORg. In some embodiments, Rg is selected from optionally substituted alkyl, optionally substituted cycloalkyl, and optionally substituted heterocycloalkyl,

[0076] In some embodiments, Z is NR10R11. In some embodiments, R10 is selected from hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl, and optionally substituted heterocycloalkyl, and Rn is selected from optionally substituted alkyl, optionally substituted cycloalkyl, and optionally substituted heterocycloalkyl. In some embodiments, R10 is hydrogen and R1 is optionally substituted alkyl. In some embodiments, R10 is hydrogen and Rn is alkyl. In some embodiments, R10 and R1 are joined together to form a 5- to 7-membered heterocycloalkyl ring.

[0077] In some embodiments, the compound of Formula I is selected from lalf compounds

<img file="ES2654584T3_D0006.tif" />

<img file="ES2654584T3_D0007.tif" />

ES 2 654 584 T3

[0078] Also provided is at least one chemical entity selected from compounds of formula II

<img file="ES2654584T3_D0008.tif" />

and pharmaceutically acceptable salts thereof, wherein

R1 and R2 are independently selected from hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl; or R1 and R2 may optionally be joined together with any intermediate atoms to form an optionally substituted heterocycloalkyl ring; for each occurrence, R3 and R4 are independently selected from hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl; or R3 and R4 may optionally be joined together with any intermediate atoms to form an optionally substituted cycloalkyl ring or an optionally substituted heterocycloalkyl ring;

or R1 and an occurrence of R3 may optionally be joined together with any intermediate atoms to form an optionally substituted heterocycloalkyl ring; and n is selected from 1, 2, 3, 4, 5, and 6.

In some embodiments, R1 and R2 are each independently chosen from hydrogen and optionally substituted alkyl group having one to six carbons. In some embodiments, R1 and R2 are hydrogen.

[0080] In some embodiments, R1 and R2 are joined together to form a 5- to 7-membered heterocycloalkyl ring.

[0081] In some embodiments, R3 and R4 are each independently chosen from hydrogen and optionally substituted alkyl group having one to six carbons.

[0082] In some embodiments, n is chosen from 1,2, and 3.

[0083] In some embodiments, n is 1, and R1 and R3 are joined together to form a 5- to 7-membered heterocycloalkyl ring.

[0084] In some embodiments, the compound of Formula II is selected from compounds II-a-II-d.

ES 2 654 584 T3

<img file="ES2654584T3_D0009.tif" />

<img file="ES2654584T3_D0010.tif" />

[0085] In some embodiments, the compound of Formula II is selected from compounds ll-and -ll-h.

<img file="ES2654584T3_D0011.tif" />

ES 2 654 584 T3

[0086] At least one chemical entity selected from compounds of Formula III is also provided

<img file="ES2654584T3_D0012.tif" />

and pharmaceutically acceptable salts thereof, wherein

Rs is selected from hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl; R6 is selected from optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted acyl, optionally substituted alkoxycarbonyl, and -P (= O) (OR7) (OR8), where R7 and Rs they are independently chosen from hydrogen and optionally substituted alkyl.

[0087] In some embodiments, R5 is selected from hydrogen and optionally substituted alkyl group having one to six carbons. In some embodiments, R5 is selected from hydrogen and alkyl group having one to six carbons. In some embodiments, R5 is selected from hydrogen and methyl.

[0088] In some embodiments, R6 is selected from optionally substituted alkyl.

[0089] In some embodiments, R6 is selected from optionally substituted acyl. In some embodiments, R6 is selected from acyl. In some embodiments, R6 is selected from acetyl, propionyl, isobutyryl, and pivaloyl.

[0090] In some embodiments, R6 is selected from optionally substituted alkoxycarbonyl. In some embodiments, R6 is selected from alkoxycarbonyl. In some embodiments, R6 is selected from optionally substituted methoxycarbonyl, ethoxycarbonyl, and isopropoxycarbonyl,

[0091] In some embodiments, R6 is selected from -P (= O) (OR7) (ORs), where R7 and Rs are independently chosen from optionally substituted hydrogen and alkyl. In some embodiments, R7 and R8 are independently chosen from the hydrogen and alkyl group having one to six carbons. In some embodiments, R6 is P (= O) (OH) (OH).

[0092] In some embodiments, the compound of formula III is selected from compounds IlI-a-III-f.

ES 2 654 584 T3

<img file="ES2654584T3_D0013.tif" />

<img file="ES2654584T3_D0014.tif" />

<img file="ES2654584T3_D0015.tif" />

[0093] The chemical entities described herein may exhibit higher solubility compared to bufalin. The solubility of the chemical entities described herein can be tested as described below. Some of the chemical entities described herein show a solubility of at least twice that of bufalin when tested under such conditions. Some of the chemical entities described herein show a solubility of at least five times that of bufalin when tested under such conditions. Some of the chemical entities described herein show a solubility of at least ten times that of bufalin when tested under such conditions.

[0094] The chemical entities described herein can be synthesized by techniques well known in the art from commercially available raw materials and the reagents used. For example, the chemical entities described herein can be prepared as illustrated below with reference to examples and reaction schemes.

[0095] Bufalin can be obtained from the skin glands of Bufo gargarizans or B. melanostictus toads and is commercially available, for example, from Slgma-Aldrlch Corp. (St. Louls, MO). Other reagents are commercially available, for example, from Slgma-Aldrlch Corp., or can be readily prepared by those skilled in the art using commonly employed synthetic methodology.

[0096] In general, compounds of Formula I can be prepared from bufalin through activated esters. Compounds of Formula II can be prepared from bufalin by standard acilation / sterilization procedures. In one approach, sterilization is carried out by reacting bufalin with the acid in the presence of a coupling agent such as DCC, EDC, or HBTU. Compounds of Formula III can be prepared from bufalin by standard alkyl / ether formation procedures. The desired product can be purified from the reaction mixture by conventional methods, for example by extraction and / or silica gel chromatography or high pressure liquid chromatography.

[0097] The chemical entities described herein can be prepared in substantially pure form, generally by standard chromatographic methods, prior to formulation into a pharmaceutically acceptable form.

[0098] The chemical entities described herein may be for use in treating a variety of cancers. The chemical entities and compositions described herein may be for use in the prevention and / or treatment of cancers, including, but not limited to, human sarcomas and carcinomas, eg, carcinomas, eg, colon carcinoma, pancreatic cancer , breast cancer, ovarian cancer, prostate cancer, fibrosarcoma, mlxosarcoma, llposarcoma, chondrosarcoma, osteogenic sarcoma, chondroma, anglosarcoma, endothellosarcoma, llnfanglosarcoma, llnfangloendothellosarcoma, slnovloma, mesothelioma, tumor,

ES 2 654 584 T3 leiomyosarcoma, rhabdomyosarcoma, squamous cell carcinoma, basal cell carcinoma, adenocarclnoma, sweat gland carcinoma, Ewing's sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinoma, medular carcinoma of the bronchus, bronchial carcinoma kidney cells, hepatoma, bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wiims tumor, cervical cancer, testicular tumor, Lung carcinoma, small cell lung carcinoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma, melanoma, rhetinoblastoma, neuroblastias for example acute lymphocytic leukemia and acute myeloid leukemia (myeloblastic, promyelocytic, myelomonocytic, monocytic, and erythroleukemia); chronic leukemia (chronic myelocytic (granulocytic) leukemia and chronic lymphocytic leukemia); and polycythemia vera, lymphoma (Hodgkin's disease and non-Hodgkin's disease), multiple myeloma, Waldenstrom's macroglobulinemia, and heavy chain disease.

[0099] In some embodiments, the chemical entities described herein are for use in treating cancers of the (i) digestive system including, without limitation, the esophagus, stomach, small intestine, colon (including colorectal), liver, and duct. intrahepatic biliary, gallbladder and other biliary, pancreatic and digestive organs;

(ii) respiratory system, including, without limitation, larynx, lung and bronchi, and other respiratory organs;

(iii) breast;

(iv) genital system, including without limitation, cervix, ovary, and prostate;

(v) urinary system, including without limitation, the urinary bladder and the renal kidney and pelvis; and (vi) oral cavity and pharynx, including without limitation, tongue, mouth, pharynx, and other oral cavity.

[0100] In some embodiments, the chemical entities described herein are for use in treating colorectal cancer, liver cancer, lung cancer, breast cancer, and oral cancer.

[0101] Chemical entities described herein that have the desired pharmacological activity can be administered, in some embodiments, as a pharmaceutically acceptable composition comprising a pharmaceutical carrier, to a patient, as described herein. Depending on the manner of introduction, the chemical entities can be formulated in a variety of ways as discussed below. The concentration of the at least one chemical entity in the formulation can range from about 0.01-100 wit.%.

[0102] Administration of the chemical entities described herein can be done in a variety of ways, including, but not limited to oral, subcutaneous, intravenous, intranasal, transdermal, intraperitoneal, intramuscular, intrapulmonary, vaginal, rectal administration , or intraocular.

[0103] Pharmaceutical dosage forms include at least one chemical entity described herein and one or more pharmaceutical excipients. As is known in the art, pharmaceutical excipients are secondary ingredients that function to activate or enhance the delivery of a drug or medicine in a variety of dosage forms (for example: oral forms such as tablets, capsules, and liquids; topical forms such as dermal, ophthalmic and otic forms; suppositories; injectables; respiratory forms and the like). Pharmaceutical excipients include inert or inactive ingredients, synergists, or chemicals that substantively contribute to the active ingredient's medicinal effects. For example, pharmaceutical excipients can function to improve flow characteristics, product uniformity, stability, taste or appearance, to facilitate handling and dosing, for convenience of use, or to control bioavailability. While pharmaceutical excipients are commonly described as inert or inactive, it is appreciated in the art that there is a relationship between the properties of pharmaceutical excipients and the dosage forms that contain them.

[0104] Pharmaceutically suitable excipients for use as carriers or diluents are well known in the art, and can be used in a variety of formulations. See, for example, Remington Pharmaceutical Sciences, 18^to editing, AR Gennaro, Editor, Mack Publishing Company (1990); Remington: The Science and Practice of Pharmacy, 21st Edition, Lippincott Williams &amp; Wilkins (2005); Handbook of Pharmaceutical Excipients, 3rd Edition, AH Kibbe, Editor, American Pharmaceutical Association and Pharmaceutical Press (2000); and Handbook of Pharmaceutical Additives, compiled by Michael and Irene Ash, Gower (1995).

[0105] Oral solid dosage forms such as tablets typically comprise one or more pharmaceutical excipients, which can for example help to impart satisfactory processing and compression characteristics, or provide additional desirable physical characteristics to the tablet. Such pharmaceutical excipients can be selected from diluents, binders, glidants, lubricants, disintegrants, colors, flavors, sweetening agents, polymers, waxes, or other solubility-retarding materials.

[0106] Compositions for intravenous administration will generally comprise intravenous fluids, that is

ES 2 654 584 T3 that is, sterile solutions of simple chemicals, such as sugars, amino acids or electrolytes, that can be easily carried through the circulatory system and assimilated.

[0107] Dosage forms for parenteral administration will generally comprise fluids, in particular intravenous fluids, that is, sterile solutions of simple chemicals, such as sugars, amino acids or electrolytes, which can be easily carried by the circulatory system and assimilated . Such fluids are typically prepared with USP Water for Injection. Commonly used fluids for intravenous (iv) use are described in Remington: The Science and Practice of Pharmacy, Lippincott Williams &amp; Wilkins (2005). The pH of such IV fluids can vary, and will typically be 3.5 to 8 as is known in the art.

[0108] The chemical entities described herein can also be used in conjunction with other well-known therapeutic agents that are selected for their particular utility against the condition being treated. For example, chemical entities described herein may be useful in combination with at least one additional anti-cancer antibody and / or cytotoxic agents. Furthermore, the chemical entities described herein may also be useful in combination with other inhibitors of parts of the signaling pathway that binds cell surface growth factor receptors to nuclear signals that initiate cell proliferation.

[0109] Such known anti-cancer and / or cytotoxic agents that can be used in combination with the chemical entities described herein include:

(i) other antiproliferative / antineoplastic drugs and combinations thereof, as used in medical oncology, such as alkylating agents (for example cis-platinum, oxaliplatin, carboplatin, cyclophosphamide, nitrogen mustard, melphalan, chlorambucil, busulfan, temozolamide and nitrosoureas); antimetabolites (for example gemcitabine and antifolates such as fluoropyrimidines such as 5-fluorouracil and tegafur, raltitrexate, methotrexate, cytosine arabinoside, and hydroxyurea); antitumor antibiotics (for example anthracyclines such as adriamycin, bleomycin, doxorubicin, daunomycin, epirubicin, idarubicin, mitomycin C, dactinomycin and mithramycin); antimitotic agents (for example vinca alkaloids such as vincristine, vinblastine, vindesine and vinorelbine and taxoids such as taxol and taxotere and polokinase inhibitors); and topoisomerase inhibitors (eg epipodophyllotoxins such as etoposide and teniposide, amsacrine, topotecan and camptothecin);

(ii) cytostatic agents such as antiestrogens (for example tamoxifen, fulvestrant, toremifene, raloxifene, droloxifene and iodoxifene), antiandrogens (for example bicalutamide, flutamide, nilutamide and cyproterone acetate), LHRH antagonists or LHRH antagonists or LHRH agonists (for example LHRHoserelonists) , leuprorelin and buserelin), progestogens (such as megestrol acetate), aromatase inhibitors (such as anastrozole, letrozole, vorazol and exemestane) and 5a reductase inhibitors such as finasteride;

(iii) anti-invasion agents [eg, inhibitors of the c-Src kinase family of type 4- (6-chloro-2,3-methylenedioxyanilino) -7- [2- (4-methylpiperazine-1-yl) ethoxy] -5-tetrahydropyran-4yloxyquinazoline (AZD0530; International Patent Application WO 01/94341), N- (2-chloro-6-methylphenyl) -2- {6- [4- (2-hydroxyethyl) piperazine-1-yl] -2-methylpyrimidine-4ylamino} thiazole-5-carboxamide (dasatinib, BMS-354825; J. Med. Chern, 2004, 47, 66586661) and bosutinib (SK1-606), and metalloproteinase inhibitors such as marimastat, urokinase inhibitors plasminogen receptor function activator or antibodies against heparanase];

(iv) inhibitors of growth factor function: for example such inhibitors include growth factor antibodies and growth factor receptor antibodies (for example the trastuzumab anti-erbB2 antibody [Herceptin^TM], anti-EGFR panitumumab antibody, anti-erbB 1 cetuximab antibody [Erbitux, C225] and any growth factor receptor antibody or growth factors described by Stem et al Critical Reviews in oncology / hematology, 2005, Vol 54, pp 11-29); Such inhibitors also include tyrosine kinase inhibitors, for example inhibitors of the epidermal growth factor family (for example inhibitors of the EGFR family tyrosine kinase, such as N- (3-chloro-4-fluorophenyl) -7- methoxy-6- (3-morpholinopropoxy) quinazoline-4-amine (gefitinib, ZD1839), N- (3-ethynylphenyl) -6,7-bis (2-methoxyethoxy) quinazoline-4amine (erlotinib, OSI-774) and 6-acrylamido N- (3-chloro-4-fluorophenyl) -7- (3-morpholinopropoxy) quinazoline-4-amine (CI 1033), erbB2 tyrosine kinase inhibitors such as lapatinib); inhibitors of the hepatocyte growth factor family; inhibitors of the insulin growth factor family; platelet-derived growth factor family inhibitors such as imatinib and / or nilotinib (AMN107); serine / threonine kinase inhibitors (eg, Ras / Raf inhibitors such as farnesyl transferase inhibitors, eg sorafenib (BAY 43-9006), tipifarnib (RI15777), and canvasfarnib (SCH66336)), cell signaling inhibitors via MEK and / or AKT kinases, c-kit inhibitors, abl kinase inhibitors, P13 kinase inhibitors, Plt3 kinase inhibitors, CSF-IR kinase inhibitors, IGF receptor (insulin-like growth factor) kinase inhibitors; aurora kinase inhibitors (eg AZD1152, PH739358, VX-680, MLN8054, R763, MP235, MP529, VX-528 and AX39459) and cyclin dependent kinase inhibitors such as CDK2 and / or CDK4 inhibitors;

(v) antiangiogenic agents such as those that inhibit the effects of vascular endothelial growth factor, [for example, anti-vascular endothelial cell growth factor antibody bevacizumab (Avastin ™) and, for example, a tyrosine kinase inhibitor of VEGF receptor such as vandetanib (ZD6474), vatalanib (PTK787), sunitinib (SU11248), axitinib (AG-013736), pazopanib (GW 786034) and 4- {4-fluoro-2-methylindole-5-yloxy) 6-methoxy -7- (3-pyrrolidine-1-ylpropoxy) quinazoline (AZD2171; Example 240 in WO 00/47212), compounds such as those described in international patent applications WO 97/22596, WO 97/30035, WO 97/32856 and WO

ES 2 654 584 T3

98/13354 and compounds that function by other mechanisms (eg linomide, av ~ 3 integrin inhibitors, and angiostatin));

(vi) vascular detrimental agents such as Combretastatin A4 and the compounds described in cations of International Patent Application WO 99/02166, WO 00/40529, WO 00/41669, WO 01/92224, WO 02/04434 and WO 02/08213 ;

(vii) an endothelin receptor antagonist, for example zibotentan (ZD4054) or atrasentan;

(viii) antisense therapies, for example those that are directed to the targets listed above, such as ISIS 2503, an anti-ras antisense;

(ix) gene therapy approaches, including for example proposals to replace aberrant genes such as aberrant p53 or aberrant BRCA1 or BRCA2, GDEPT (gene-directed enzyme prodrug therapy) such as those using cytosine deaminase, thymidine kinase or a bacterial nitroreductase enzyme and approaches to increasing tolerance subject to chemotherapy or radiation therapy, such as multidrug resistance gene therapy; and (x) immunotherapy approaches, including for example ex-vivo and in-vivo to increase the immunogenicity of the subject's tumor cells, such as transfection with cytokines such as interleukin 2, interleukin 4, or granulocyte macrophage colony-stimulating factor. , approaches to lowering T cell anergy, approaches using transfected immune cells such as dendritic cells transfected with cytokines, strategies using cytokine-transfected tumor cell lines and approaches using anti-idiotypic antibodies.

[0110] In certain embodiments, the at least one chemical entity is administered in combination with one or more agents selected from pacliataxel, bortezomib, dacarbazine, gemcitabine, trastuzumab, bevacizumab, capecitabine, docetaxel, erlotinib, aromatase inhibitors, such as AROMASIN ™ (exemestane), and estrogen receptor inhibitors, such as FASLODEX ™ (fulvestrant).

[0111] When a chemical entity described herein is administered to a human subject, the daily dosage will normally be determined by the prescribing physician with the dosage generally varying according to the age, weight and response of the individual subject, as well as the severity of the subject's symptoms.

[0112] In an exemplary application, a suitable amount of at least one chemical entity is administered to a mammal undergoing treatment for cancer, eg, breast cancer. Administration normally occurs in an amount of between about 0.01 mg / kg of body weight to about 100 mg / kg of body weight per day (administered in single or divided doses), such as at least about 0.1 mg / kg of body weight per day. A particular therapeutic dose may include, for example, from about 0.01 mg to about 1000 mg of the chemical entity, such as including, for example, from about 1 mg to about 1000 mg. The amount of the entity at least one chemical in a unit dose of preparation can be varied or adjusted from about 0.1 mg to 1000 mg, such as from about 1 mg to 300 mg, for example 10 mg to 200 mg, according to the particular request. The amount administered will vary depending on the particular IC50 value of the at least one chemical entity used and the judgment of the attending physician taking into consideration factors such as health, weight, and age. In combinational applications where the at least one chemical entity described herein is not the only active ingredient, it may be possible to administer lower amounts of the at least one chemical entity and still have a therapeutic or prophylactic effect.

[0113] In some embodiments, the pharmaceutical preparation is in unit dosage form. In such form, the preparation is subdivided into unit doses containing appropriate amounts of the active component, eg, an effective amount to achieve the desired purpose.

[0114] The actual dosage employed can be varied depending on the requirements of the subject and the severity of the condition to be treated. Determination of the appropriate dosage for a particular situation is within the skill of the art. Generally, treatment is started with smaller dosages that are less than the optimal dose of the at least one chemical entity. Thereafter, the dosage is increased in small amounts until the optimal effect is reached under the circumstances. For convenience, the total daily dosage can be divided and administered in portions throughout the day if desired.

[0115] The amount and frequency of administration of the at least one chemical entity described herein, and whether other applicable chemotherapeutic agents and / or radiation therapy, will be regulated according to the judgment of the attending physician considering factors such as age, condition and size of the subject, as well as the severity of the disease to be treated.

[0116] The chemotherapeutic agent and / or radiation therapy can be administered according to therapeutic protocols well known in the art. It will be apparent to those skilled in the art that the administration of the chemotherapeutic agent and / or radiation therapy can be varied depending on the disease being treated and the known effects of the chemotherapeutic agent and / or radiation therapy on that disease. Also, in accordance with the knowledge of the skilled practitioner, therapeutic protocols (eg, dosage amounts and times of administration) may be varied in view of the observed effects of the agents.

ES 2 654 584 T3 administered therapeutics (ie, antineoplastic agent or radiation) on the subject, and in view of observed disease responses to administered therapeutic agents.

[0117] Furthermore, in general, the at least one chemical entity described herein need not be administered in the same pharmaceutical composition as a chemotherapeutic agent, and may, due to different physical and chemical characteristics, be administered by a different route. For example, the chemical entities / compositions can be administered orally to generate and maintain good blood levels thereof, while the chemotherapeutic agent can be administered intravenously. Determination of the mode of administration and the convenience of administration, when possible, in the same pharmaceutical composition, is well within the knowledge of the skilled practitioner. The initial administration can be done according to established protocols known in the art and then, based on the observed effects, the dosage, modes of administration and times of administration can be modified by the skilled clinician.

[0118] The particular choice of chemical entity (and where appropriate, chemotherapeutic agent and / or radiation) will depend on the diagnosis of the attending physicians and their judgment of the subject's condition and the appropriate treatment protocol.

[0119] The chemical entities described in this document (and where appropriate chemotherapeutic agent and / or radiation) can be administered concurrently (for example, simultaneously, essentially simultaneously or within the same treatment protocol) or sequentially, depending on the nature of the proliferative disease, the condition of the subject, and the actual choice of chemotherapeutic agent and / or radiation to be co-administered (i.e., within a single treatment protocol) with the entity / chemical composition.

[0120] In combinational uses and applications, the chemical entity / composition and the chemotherapeutic agent and / or radiation do not have to be administered simultaneously or essentially simultaneously, and the initial order of administration of the chemical entity / composition, and the chemotherapeutic agent and / or radiation, may not be important. Therefore, the at least one chemical entity described herein can be administered first followed by administration of the chemotherapeutic agent and / or radiation; or the chemotherapeutic agent and / or radiation may be administered first followed by the administration of the at least one chemical entity described herein. This alternative administration can be repeated during a single treatment protocol. Determination of the order of administration, and the number of repetitions of administration of each therapeutic agent during a treatment protocol, is well within the knowledge of the skilled practitioner after evaluation of the disease to be treated and the condition of the subject. For example, the chemotherapeutic agent and / or radiation may be administered first, and then treatment continued with the administration of the at least one chemical entity described herein, followed, where determined advantageous, by the administration of the chemotherapeutic agent and / or radiation, and so on until the treatment protocol has been completed.

[0121] Thus, according to experience and knowledge, the practicing physician can modify each protocol for the administration of an entity / chemical composition for treatment according to the needs of the individual subject, such as the treatment product.

[0122] The attending physician, in judging whether the treatment is effective at the administered dosage, will consider the general well-being of the subject, as well as more defined signs such as alleviation of disease-related symptoms, inhibition of tumor growth , the actual shrinkage of the tumor, or the inhibition of metastasis. Tumor size can be measured by standard methods such as radiological studies, eg CAT or MRI scan, and successive measurements can be used to judge whether or not tumor growth has been retarded or even reversed. Relief of disease-related symptoms such as pain, and improvement in general condition can also be used to help judge the effectiveness of treatment.

EXAMPLES

[0123] The following examples serve to more fully describe how to use the invention. These examples are presented for illustrative purposes.

[0124] In performing the procedures of the methods described herein, it should, of course, be understood that reference to certain buffers, media, reagents, cells, culture conditions, and the like are not intended to be limiting, but rather they should be read to include all related materials that one of ordinary skill in the art would recognize as of interest or value in the particular context in which that discussion is presented. For example, it is often possible to substitute one buffer system or culture medium for another and still achieve similar, if not identical, results. Those skilled in the art will have sufficient knowledge of such systems and methodologies that being able, without undue experimentation, to make such substitutions as optimally will serve their purposes in using the methods and procedures described herein.

ES 2 654 584 T3

Example I: Preparation of (R) - (3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxy-10,13-dimethyl-17- (2-oxo-2H-pyran-5yl) hexadecahydro-1H-cyclopenta [a] phenanthrene-3-yl 2-aminopropanoate

[0125]

<img file="ES2654584T3_D0016.tif" />

(R) - (3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-h id roxy-10,13-dim ethyl-17- (2-oxo-2H-plran-5o ) hexadecahldro-1/7-cyclopenta [a] phenanthrene-3-llo 2-aminepropanoate

<img file="ES2654584T3_D0017.tif" />

BocHN

<img file="ES2654584T3_D0018.tif" />

[0126] To a solution of Boc-amine acid (11.3 mg, 0.06 mmol, 1.2 eq), HOBT (9.7 mg, 0.072 mmol, 1.44 eq), EDC (13.8 mg , 0.072 mmol, 1.44 eq) and DMAP (16.8 mg, 0.15 mmol, 3 eq) in CH2Cl2 was added bufalin (20 mg, 0.05 mmol). The mixture was stirred at 37 ° C for 16 h and then purified by preparative TLC (PE / EA = 1: 1) to provide (R) - (3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) - 14-hydroxl-10,13-dimetho-17- (2-oxo-2H-pyran-5ylo) hexadecahldro-1H-cyclopenta [a] phenanthren-3-lo 2- ( (tert-butoxycarbonylo) amine) propanoate (23 mg, 79.8%).

BocHN

<img file="ES2654584T3_D0019.tif" />

<img file="ES2654584T3_D0020.tif" />

[0127] To a solution of (R) - (3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxl-10,13-dimetho-17- (2-oxo- 2H-pyran-5lo) hexadecahydro-1H-cyclopenta [a] phenanthren-3-lo 2 - ((tert-butoxycarbonyl) amino) propanoate in EtOAc (3 mL) HCl (4 M in EtOAc, 3 mL) dropwise at 0 ° C. The resulting mixture was warmed to rt after 30 min and stirred for 2 h. The mixture was quenched with saturated NaHCO3 solution and extracted with EtOAc (20 mL X 3). The organic layer was washed with H2O (10 mL X 4) and then dried over anhydrous Na2SC&gt; 4, concentrated under reduced pressure. The crude product was then purified via prep-TLC (CH2Cl2: MeOH = 10: 1) to provide the (R) - (3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydrox¡- 10,13-dimethyl-17- (2-oxo-2H-pyran-5-llo) hexadecahydro-1Hclclopenta [a] phenanthren-3-llo 2-aminepropanoate (8 mg, 43% of yield) as a white solid. LRMS (M + H⁺) m / z 458.5. 1H NMR (CD3OD, 400 MHz)? 7.89 (dd, J = 9.6, 2.4 Hz, 1H), 7.3 3 (m, 1H), 6.17 (d, J = 9.6 Hz, 1H), 5.02 ( m, 1H), 3.54 (m, 1H), 2.43-2.48 (m, 1H), 1.8-2.15 (m, 24H), 0.88 (s, 3H), 0 , 62 (s, 3H).

ES 2 654 584 T3

Example II: Preparation of (3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxy-10,13-dimethyl-17- (2-oxo-2H-pyran-5-yl) hexadecahydro-1H- cyclopenta [a] phenanthren-3-yl (2- (pyrrolidine-1-yl) ethyl) carbonate

[0128]

<img file="ES2654584T3_D0021.tif" />

(3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxy-10,13-dimethyl-17- (2-oxo-2H-pyran-5-yl) hexadecahydro-1H-cyclopenta [a] phenanthren -3-yl (2- (pyrrolidine-1-yl) ethyl) carbonate

[0129] To a solution of 1 (60 mg, 0.15 mmol), and DMAP (16.8 mg, 0.15 mmol) in CH2CL (10 ml) was added DIEA (77.4 mg, 0.6 mmol ) and 4-nitrophenyl chlorocarbonate (60.6 mg, 0.3 mmol). The mixture was stirred at 37 ° C for 16 h and then purified by preparative TLC (PE / eA = 1: 1) to provide (3S, 5R, 8R, 9S, 110S, 13R, 14S, 17R) -14-hydroxy- 4-nitrophenyl 10,13-dimethyl-17- (2-oxo-2H-pyran-5-yl) hexadecahydro-1H-ichlopenta [a] phenanthren-3-yl carbonate as a white solid (72 mg, 87.1 %).

<img file="ES2654584T3_D0022.tif" />

[0130] To a solution of (3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxy-10,13-dimethyl-17- (2-oxo-2H-pyran-5yl) hexadecahydro- 1H-cyclopenta [a] phenanthren-3-yl carbonate of 4-nitrophenyl (24 mg, 0.044 mmol) in CH2CL was added 2- (pyrrolidine-1-yl) ethanol (50.6mg, 0.44 mmol, 10 eq) , DIEA (22.7 mg, 0.176 mmol, 4 eq) and DmAP (19.7 mg, 0.176 mmol, 4 eq). The resulting mixture was stirred at 40 ° C for 16 h and then purified by preparative TLC to provide (3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxy-10,13-dimethyl-17- (2-oxo-2H-pyran-5yl) hexadecahydro-1H-cyclopenta [a] phenanthren-3-yl 2- (pyrrolidine-1-yl) ethyl carbonate (20mg, 87.0%) as a white solid. LRMS (M + H +) m / z 528.4. 1H NMR (CD3O D, 400 MHz)? 8.01 (dd, J = 10.0, 2.4 Hz, 1H), 7.44 (m, 1H), 6.29 (d, J = 10.0 Hz, 1H), 5.00 (m , 1H), 4.35 (t, J = 5.4 Hz,²H), 3.12 (m, 2H), 2.96 (m, 4H), 2.55-2.60 (m, 1H), 1.08-2.15 (m, 25H), 0.99 (s, 3H), 0.73 (s, 3H).

Example III: Preparation of acid 4 - (((((3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxy-10,13-dimethyl-17- (2-oxo2H-pyran-5 -yl) hexadecahydro-1H-cyclopenta [a] phenanthren-3-yl) oxy) carbonyl) amino) butanoic

[0131]

ES 2 654 584 T3

<img file="ES2654584T3_D0023.tif" />

Acid 4 - (((((3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxy-10,13-dimethyl-17- (2-oxo-2H-pyran-5-yl) hexadecahydro- 1H-cyclopenta [a] phenanthren-3-yl) oxy) carbonyl) amino) butanoic

<img file="ES2654584T3_D0024.tif" />

[0132] To a solution of (3S, SR, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxy-10,13-dimethyl-17- (2-oxo-2H-pyran-5yl) hexadecahydro- 1H-cyclopenta [a] phenanthren-3-yl carbonate of 4-nitrophenyl (20 mg, 0.036 mmol) in CH2Cl2 was added 4-aminobutanoic acid (37.1 mg, 0.36 mmol, 10 eq), DIEA (18, 6 mg, 0.144 mmol, 4 eq) and DMAP (16.1 mg, 0.144 mmol, 4 eq). The resulting mixture was stirred at 40 ° C for 16 h and then purified by preparative TLC to provide acid 4 - (((3S, 5R, 8R, 9S, 10S, 13R, 145.17R) -14-hydroxy-10.13 -dimethyl-17- (2-oxo2H-pyran-5-yl) hexadecahydro-1H-cyclopenta [a] phenanthren-3-yloxy) carbonylamino) butanoic (10mg, 53.5%) as a white solid. LRMS (MH +) m / z 514.4.¹H NMR (CD3OD, 400 MHz) δ 7.89 (dd, J = 9.6, 2.4 Hz, 1H), 7.3 3 (m, 1H), 6.18 (d, J = 9.6 Hz, 1H), 4.82 (m, 1H), 3.03 (m, 2H), 2.44-2.48 (m, 1H), 1.08-2.15 (m, 25H), 0 , 87 (s, 3H), 0.62 (s, 3H).

Example IV: Preparation of (3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxy-10,13-dimethyl-17- (2-oxo-2H-pyran-5-yl) hexadecahydro-1H- cyclopenta [a] phenanthren-3-yl (2- (pyrrolidine-1-yl) ethyl) carbamate

[0133]

ES 2 654 584 T3

<img file="ES2654584T3_D0025.tif" />

(3 S, 5R, 8R, 9 S, 10 S, 13R, 14 S, 17R) -14-hydroxy-10,13-dimethyl-17- (2-oxo-2 H-pyran-5-yl) hexadecahydro-1H- cyclopenta [a] phenanthren-3-yl (2- (pyrrolidine-1-yl) ethyl) carbamate

<img file="ES2654584T3_D0026.tif" />

[0134] To a solution of (3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxy-10,13-dimethyl-17- (2-oxo-2H-pyran-5-yl) hexadecahydro- 4-Nitrophenyl 1H-cyclopenta [a] phenanthren-3-yl carbonate (29 mg 0.054 mmol) was added in CH2Cl2 2- (pyrrolidine-1-yl) ethanamine (61.6 mg, 0.54 mmol). The resulting mixture was stirred at rt for 16 h and then purified by preparative TLC to provide (3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxy10,13-dimethyl-17- (2-oxo Methyl -2H-pyran-5-yl) hexadecahydro-1H-cyclopenta [a] phenanthren-3-yl (2- (pyrrolidine-1-yl) ethyl) carbamate (21 mg, 75%) as a white solid. LRMS (M + H +) m / z 527.5.¹H NMR (CD3OD 400 MHz) δ 7.89 (dd, J = 9.6, 2.4 Hz, 1H), 7.33 (m, 1H), 6.18 (d, J = 9.6 Hz, 1H), 4.88 (m, 1H), 3.35 (m, 2H), 3.12 (m, 2H), 2.46 (m, 1H), 1.08-2.15 (m, 29H ), 0.87 (s, 3H), 0.62 (s, 3H).

Example V: Preparation of (3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxy-10,13-dimethyl-17- (2-oxo-2H-pyran-5-yl) hexadecahydro-1H- cyclopenta [a] phenanthren-3-yl piperazine-1-carboxylate

[0135]

<img file="ES2654584T3_D0027.tif" />

(3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxy-10,13-dimethyl-17- (2-oxo-2H-pyran-5-yl) hexadecahydro-1H-cyclopenta [a] phenanthren -3-yl piperazine-1-carbamate

<img file="ES2654584T3_D0028.tif" />

ES 2 654 584 T3

[0136] To a solution of 3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxy-10,13-dimethlo-17- (2-oxo-2H- p¡ran-5¡lo) hexadecahydro-1H-cyclopenta [a] phenanthren-3-¡lo was added 4-nitrophene carbonate (29 mg, 0.054 mmol) in CH2Cl2 p¡ perazine (46.4 mg, 0.54 mmol). The resulting mixture was stirred at rt for 16 h and then purified by preparative TLC to provide (3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-h¡ drox¡-10,13-d¡ methyl-17- (2oxo-2H-p¡ran-5-¡lo) hexadecahydro-1H-cyclopenta [a] phenanthren-3-¡lo p¡peraz Na-1-carboxylate (18.6 mg, 69.2%) as a white solid. LRMS (M + H +) m / z 499.5.¹H NMR (CD3OD, 400 MHz)? 7.90 (dd, J = 9.6, 2.4 Hz, 1H), 7.33 (m, 1H), 6.18 (d, J = 9.6 Hz, 1H), 4.89 (m , 1H), 3.41 (m, 4H), 2.77-2.80 (m, 4H), 2.44-2.48 (m, 1H), 1.08-2.15 (m, 21H ), 0.88 (s, 3H), 0.62 (s, 3H).

Example VI: Preparation of (3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydrox¡ -10,13-d¡meto-17- (2-oxo-2H -p¡ran-5¡lo) hexadecahydro-1H-cyclopenta [a] phenanthren-3-¡lo (2-morpholnoet¡lo) carbamate

[0137]

<img file="ES2654584T3_D0029.tif" />

(3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydrox¡ -10,13-d¡melo-17- (2-oxo-2H-p¡ran-5¡ lo) hexadecahydro-1H-cyclopenta [a] phenanthren-3-lo (2-morpholnoethyl) -carbamate

<img file="ES2654584T3_D0030.tif" />

[0138] To a solution of (3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxy-10,13-dimethlo-17- (2-oxo -2H-pyran-5lo) hexadecahydro-1H-cyclopenta [a] phenanthren-3-lo was added 4-nitrophene carbonate (29 mg, 0.054 mmol) in CH2Cl2 2- morphol¡noethanam¡ne (70.2 mg, 0.54 mmol). The resulting mixture was stirred at rt for 16 h and then purified by preparative TLC to provide (3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-h¡ drox¡-10,13d¡met¡lo-17- (2-oxo-2H-pyran-5-¡lo) hexadecahydro-1H-cyclopenta [a] phenanthren-3-¡lo (2- morpholinoethyl) carbamate (18 mg, 61.4%) as a white solid. LRMS (M + H +) m / z 543.4.¹H NMR (CD3OD, 400 MHz) δ 7.89 (dd, J = 9.6, 2.4 Hz, 1H), 7.33 (m, 1H), 6.18 (d, J = 9.6 Hz , 1H), 4.82 (m, 1H), 3.59 (m, 4H), 3.16 (m, 2H), 2.46 (m, 2H), 2.41 (m, 5H), 1 , 08-2.15 (m, 21H), 0.87 (s, 3H), 0.61 (s, 3H).

Example VII: Additional compounds

[0139] Using methods similar to those described above, the following compounds were also prepared.

ES 2 654 584 T3

<td>Chemical name</td><td>Ion</td><td>m / z observed</td>

<td>(R) - (3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxy-10,13-dimethyl-17- (2-oxo-2Hpyran-5-yl) hexadecahydro-1H-cyclopenta [a] phenanthren-3-yl 2-amino-3- methylbutanoate</td><td>M + H +</td><td> 486,5</td>

<td>(R) - (3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxy-10,13-dimethyl-17- (2-oxo-2Hpyran-5-yl) hexadecahydro-1H-cyclopenta [a] phenanthren-3-yl 2-amino-4- methylpentanoate</td><td>M + H +</td><td> 500,6</td>

<td>(S) - (3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxy-10,13-dimethyl-17- (2-oxo-2Hpyran-5-yl) hexadecahydro-1H-cyclopenta [a] phenanthren-3-yl 2-amino-4- methylpentanoate</td><td>M + H +</td><td> 500,5</td>

<td>(3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxy-10,13-dimethyl-17- (2-oxo-2H-pyran-5-yl) hexadecahydro-1H-cyclopenta [a] phenanthren -3-yl (2- morpholinoethyl) carbonate</td><td>M + H +</td><td> 544,5</td>

<td>(S) - (3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxy-10,13-dimethyl-17- (2-oxo-2Hpyran-5-yl) hexadecahydro-1H-cyclopenta [a] phenanthren-3-yl 2-aminopropanoate</td><td>M + H +</td><td> 458,5</td>

<td>(S) - (3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxy-10,13-dimethyl-17- (2-oxo-2Hpyran-5-yl) hexadecahydro-1H-cyclopenta [a] phenanthren-3-yl 2-amino-3- methylbutanoate</td><td>M + H +</td><td> 486,5</td>

<td>(3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxy-10,13-dimethyl-17- (2-oxo-2H-pyran-5-yl) hexa-decahydro-1H-cyclopenta [a ] phenanthren-3-yl morpholine-4- methyl carboxylate</td><td>M + H +</td><td> 500,4</td>

Example VIII: Equilibrium solubility measurement.

[0140] The equilibrium solubility of the compounds is measured in aqueous buffer. An excess amount of solid compound in buffer solution was added and the sample was sonicated briefly and then stirred at rt for 24 h. The sample is filtered and the concentration is analyzed by UV HPLC. A 0.2 mg / ml standard solution was prepared in methanol or acetonitrile for each compound and is used as an external standard for quantification. Data for bufalin and four of the compounds specifically described herein in NaOAc / AcOH buffer (100 mM, pH 5.0) are shown below.

<td>Chemical name</td><td>Solubility (mg / mL)</td>

<td>Bufalina</td><td> 0,041</td>

<td>(S) - (3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxy-10,13-dimethyl-17- (2-oxo-2H-pyran-5-yl) hexadecahydro-1H-cyclopenta [a] phenanthren-3-yl 2-amino-3-methylbutanoate</td><td> 0,12</td>

<td>(3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxy-10,13-dimethyl-17- (2-oxo-2H-pyran-5-yl) hexadecahydro-1H-cyclopenta [a] phenanthren -3-yl (2- (pyrrolidine-1-yl) ethyl) carbonate</td><td> 0,71</td>

<td>(3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxy-10,13-dimethyl-17- (2-oxo-2H-pyran-5-yl) hexadecahydro-1H-cyclopenta [a] phenanthren -3-yl (2- (pyrrolidine-1-yl) ethyl) carbamate</td><td> 1,04</td>

<td>(3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxy-10,13-dimethyl-17- (2-oxo-2H-pyran-5-yl) hexadecahydro-1H-cyclopenta [a] phenanthren -3-yl piperazine-1-carboxylate</td><td> 0,84</td>

Example IX: Inhibition of cell growth in tumor cells.

[0141] Inhibition of cell growth by the compounds was measured using the MTT assay (Mosmann, T., Journal of Immunological Methods, 1983, 65, 55-63). Tumor cell lines were purchased from ATCC (American Type Culture Collection, Manassas, VA). All cell lines were maintained in RPMI 1640 (Hyclone) supplemented with 10% fetal bovine serum (FBS, hyclone), glutamine (2 mM, hyclone) and antibiotics (100 U / ml penicillin and 50 mg / ml streptomycin) at 37 ° C in a humidified atmosphere of 5% CO2 in air. Taxol (positive control, Sigma) and compounds were dissolved in DMSO (Sigma), and the final concentration of DMSO in the medium was 1%. Tumor cells were seeded in 96-well plates at densities of 4000 cells / well of a 96-well plate and allowed to adhere / grow for 24 h. They were then treated with various concentrations of drug for 72 h. 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium (MTT, Sigma) was used to determine the number of viable cells at the time of compound addition and the number of cells remaining after the exposure to the compound for 72 h. The number of cells remaining after 72 h was compared with the number of viable cells at the time of compound addition by measuring the absorbance at 570 nm, allowing the calculation of growth inhibition.

[0142] All compound concentrations were tested in triplicate and controls were averaged over 4 wells. IC50 was calculated by plotting compound concentration vs. percent inhibition in treated wells using GraphPad Prism 5. Data for representative buffalo and compounds are shown below.

ES 2 654 584 T3

Table I. The inhibitory activity of representative compounds in A549 cells.

<td>Chemical name</td><td>Cell A549 IC50 (nM)</td>

<td>Bufalina</td><td> 4,4</td>

<td>(R) - (3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxy-10,13-dimethyl-1 7- (2-oxo-2H-pyran-5yl) hexadecahydro-1H- cyclopenta [a] phenanthrin-3-yl 2-aminopropanoate</td><td> 3,4</td>

<td>(R) - (3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxy-10,13-dimethyl-1 7- (2-oxo-2H-pyran-5yl) hexadecahydro-1H- cyclopenta [a] phenanthrin-3-yl 2-amino-3-methylobutanoate</td><td> 147,1</td>

<td>(R) - (3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxy-10,13-dimethyl-1 7- (2-oxo-2H-pyran-5yl) hexadecahydro-1H- cyclopenta [a] phenanthrin-3-yl 2-amino-4-methylopentanoate</td><td> 58,8</td>

<td>(S) - (3S, 5R, 8R, 9S, 10S, 13R, 145,17R) -14-hydroxy-10,13-dimethyl-1 7- (2-oxo-2H-pyran-5yl) hexadecahydro-1H- cyclopenta [a] phenanthrin-3-yl 2-amino-4-methylopentanoate</td><td> 15,1</td>

<td>(3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxy-10,13-dimethyl-17- (2-oxo-2H-pyran-5-yl) hexadecahydro-1H-cyclopenta [a] phenanthrin -3-yl (2-morpholinoethyl) carbonate</td><td> 12,2</td>

<td>(3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxy-10,13-dimethyl-17- (2-oxo-2H-pyran-5-yl) hexadecahydro-1H-cyclopenta [a] phenanthrin -3-yl (2- (pyrrolidine-1-yl) ethyl) carbamate</td><td> 2,8</td>

<td>(3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxy-10,13-dimethyl-17- (2-oxo-2H-pyran-5- yl) hexadeca-hydro-1H-cyclopenta [a] phenanthrin-3-yl (2-morpholino-ethyl) -carbamate</td><td> 7,4</td>

<td>(3S, 5R, 8R, 9S, 10S, 13R, 145.17R) -14-hydroxy-10,13-dimethyl-17- (2-oxo-2H-pyran-5-yl) hexadeca-hydro-1H-cyclopenta [a ] phenanthrin-3-yl piperazine-1-carboxylate</td><td> 1,8</td>

<td>Acid 4 - (((((3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxy-10,13-dimethyl-17- (2-oxo-2H- pyran-5-yl) -hexadecahydro-1H-cyclopenta [a] phenanthrin-3-yl) - oxy) carbonyl) amino) butanoic</td><td> 60,0</td>

Table II. The inhibitory activity of representative compounds on Bcap-37 cells.

<td>Chemical name</td><td>Bcap37 IC50 cell (nM)</td>

<td>Bufalina</td><td> 14,0</td>

<td>(S) - (3S, 5R, 8R, 9S, 10S, 13R, 145,17R) -14-hydroxy-10,13-dimethyl-1 7- (2-oxo-2Hpyran-5-yl) hexadecahydro-1H- cyclopenta [a] phenanthrin-3-yl 2-aminopropanoate</td><td> 15,9</td>

<td>(S) - (3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxy-10,13-dimethyl-1 7- (2-oxo-2H-pyran-5-yl) hexadecahydro-1H- cyclopenta [a] phenanthrin-3-yl 2-amino-3-methylbutanoate</td><td> 48,9</td>

<td>(3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxy-10,13-dimethyl-17- (2-oxo-2H-pyran-5-yl) hexadecahydro-1H-cyclopenta [a ] phenanthren-3-yl (2- (pyrrolidine-1-yl) ethyl) carbonate</td><td> 10,6</td>

<td>(3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxy-10,13-dimethyl-17- (2-oxo-2H-pyran-5-yl) hexa-decahydro-1H-cyclopenta [a ] phenanthren-3-yl (2- (pyrrolidine-1-yl) ethyl) carbamate</td><td> 5,3</td>

<td>(3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R) -14-hydroxy-10,13-dimethyl-17- (2-oxo-2H-pyran-5-yl) hexa-decahydro-1H-cyclopenta [a ] phenanthren-3-yl morpholine-4-carboxylate</td><td> 223,0</td>

Example X: Inhibition of tumor growth in the xenograft model.

[0143] Cells were implanted into female BALB / c nude mice and grown as tumor xenografts. When tumors reached 150 to 200 mm³, mice were assigned to treatment and control groups using the random block design based on their tumor volumes. Each group contained 10 tumor-bearing mice. Tumors were measured twice a week in two dimensions using a caliper, and tumor volume was calculated from two-dimensional measurements using the equation V = 0.5 xa xb² where a and b are the long and short diameters of the tumor, respectively. Tumor volume was then used for calculations of T / C values. The T / C value was an indication of antitumor efficacy; T and C were the mean volume of the treated and control groups, respectively, on a given day. The data for one of the compounds specifically described in Example IX is shown below.

<td></td><td>Dose (mg / kg)</td><td>Schedule</td><td>Via</td><td>Tumor volume pretreatment (mm³)</td><td>Post-treatment tumor volume (mm³)</td><td>T / C</td>

<td>Vehicle</td><td> --</td><td>QDX10</td><td>iv</td><td> 151 ± 12</td><td> 524 ± 53</td><td> --</td>

<td>Compound</td><td> 3</td><td>QDX10</td><td>iv</td><td> 151 ± 12</td><td> 261 ± 26</td><td> 49,8%</td>

<td>Paclitaxel</td><td> 10</td><td>Q4DX3</td><td>iv</td><td> 152 ± 13</td><td> 391 ± 43</td><td> 74,6%</td>