WO2012097196A1 - Pyrazolopyrimidine derivatives and uses as anticancer agents - Google Patents

Abstract

Novel compounds having a fused pyrazolopyrimidine derivatives and related fused ring systems are disclosed. The compounds inhibit growth of a variety of types of cancer cells, and are thus useful for treating cancer. Pharmaceutical compositions, and methods of using these compounds and compositions to treat cancer and other diseases related to the dysregulation of kinase (such as Aurora A, Aurora B, Aurora C, cMet, JAK2, ROS1, but not limited to) pathways are disclosed. Efficacy of these compounds is demonstrated with a system for monitoring cell growth/migration, which shows they are potent inhibitors of growth and/or migration of cancer cells. Compositions comprising these compounds, and methods to use these compounds and compositions for treatment of cancers, are disclosed.

Description

PYRAZOLOPYRIMIDINE DERIVATIVES AND USES AS

ANTICANCER AGENTS

Cross-reference to related applications

[0001] This application claims priority to U.S. Provisional Patent Application Nos.

61/432,164, filed January 12, 2011, the disclosure of which is hereby incorporated herein by reference in their entirety.

Field of the Invention

[0002] The field of this invention is pharmaceutical compounds, compositions and methods, especially as they are related to compositions and methods for the treatment of cancer and other diseases related to the dysregulation of kinase (such as Aurora A, Aurora B, Aurora C, cMet, JAK2, ROS1, but not limited to) pathways.

Background of the Invention

[0003] Protein kinases are a group of enzymes that regulate diverse, important biological processes including cell growth, proliferation, survival, invasion and differentiation, organ formation, tissue repair and regeneration, etc. Protein kinases exert their physiological functions through catalyzing the phosphorylation of protein and thereby modulating the cellular activities. Because protein kinases have profound effects on cells, their activities are highly regulated. Kinases are turned on or off by phosphorylation (sometimes by

autophosphorylation), by binding of activator proteins or inhibitor proteins, or small molecules, or by controlling their location in the cell relative to their substrates. Dysfunctions in the activities of kinases, arising from genetic abnormalities or environmental factors, are known to be associated with many diseases. Several severe pathological states, including cancer and chronic inflammation, are associated with stimulation of intra-cellular signaling, and since kinases positively relay signaling events, their inhibition offers a powerful way to inhibit or control signal transduction cascades.

[0004] The proto-oncogene cMet is a member of a distinct subfamily of heterodimeric receptor tyrosine kinases which include cMet, Ron and Sea. cMet is the cell surface receptor for hepatocyte growth factor (HGF, also known as scatter factor) that conveys a unique combination of pro-migratory, anti-apoptotic and mitogenic signals. Inappropriate activation of cMet can be induced by specific genetic lesions, transcriptional upregulation or ligand- dependent autocrine or paracrine mechanisms. Amplification of the MET gene, with consequent protein overexpression and constitutive kinase activation, has been reported in a number of human primary tumors, including non-small-cell lung (NSCL) carcinomas, gastric and esophageal carcinomas with acquired resistance to epidermal growth factor receptor (EGFR) inhibitors and medulloblastomas. The MET gene can also carry activating mutations, such as those found in a subset of patients with hereditary and sporadic papillary renal cancer, gastric carcinoma and childhood hepatocellular carcinoma. HGF can also aberrantly activate cMet in an autocrine fashion, as described for glioblastomas, breast carcinomas, osteosarcomas and rhabdomyosarcomas. (Comoglio, P. M., et ah, Nature Reviews Drug Discovery (2008) 7:504- 516) Therefore cMet inhibitors may be useful in treating diseases such as cancer and other disease related to abnormal cell growth and cMet activation.

Summary of the Inventions

[0005] The present invention is directed to various classes of fused pyrazolopyrimidine derivatives and related fused ring systems described herein, pharmaceutical compositions, and methods of using these compounds and compositions to treat cancer and other diseases related to the dysregulation of kinase (such as Aurora A, Aurora B, Aurora C, cMet, JAK2, ROSl, but not limited to) pathways. These compounds have been shown as described herein to possess inhibition to protein kinases including receptor tyrosine kinases such as those of the MET subfamily, and have anti-cancer activity in cell based assays using MET addicted cell lines, which demonstrate antiproliferative and anti-migration activity against such cells. Accordingly, the compounds and compositions comprising the compounds of the invention are useful to treat conditions characterized by undesired cell proliferation and migration. In particular, the compounds are useful to treat the following type of cancer, but not limited to: carcinomas {e.g. , bladder, breast, cervical, ovarian, gastric, colorectal, esophageal, head and neck, lung, kidney, liver, cholangiocarcinoma, nasopharyngeal, pancreas, prostate, thyroid), soft tissue sarcomas {e.g. , fibrosarcoma, leiomyosarcoma, Kaposi's sarcoma), musculoskeletal sarcomas {e.g. , rhabdomyosarcoma, osteosarcoma, synovial sarcoma), hematopoietic malignancies {e.g. , multiple myeloma, lymphomas, acute myelogenous leukemia, chronic myeloid leukemia, adult T cell leukemia), and other neoplasms {e.g. , glioblastomas, melanoma, mesothelioma, astrocytomas and Wilms' tumor).

[0006] The pyrazolopyrimidine or similar heterocyclic moiety of the compounds described herein can be further fused with other aryl/ non-aryl ring or substituted with substituted aryl amino, substituted arylthio, substituted aryloxy, substituted heterocyclic amino, substituted heterocyclic thio, and substituted heterocyclic oxy derivatives thereof. The compounds as described herein exhibit anti-tumor, anticancer, anti-inflammation, anti-infectious, and anti- proliferation activity. The present invention also relates to the methods of making and formulating the described compounds and methods to use them therapeutically.

[0007] In one aspect, the contemplated heterocyclic compounds have a structure according to Formula la and lb:

Formula la Formula lb where in a ring indicates the ring is an aromatic or heteroaromatic ring;

W¹, W² W³ _, W⁴ are each independently absent, N, NH, NR¹, O, S, CH, or CR²,

provided at least one of W¹, W² W³ _, and W⁴ is CH or CR²,

not more than one of them is absent,

and not more than one of them is O or S;

R' is Ci_8 alkyl, C₂-8 alkenyl, C₂-8 alkynyl, aryl, or heteroaryl, wherein each alkyl, alkenyl, alkynyl, aryl and heteroaryl is optionally substituted;

Z is S, SO, S0₂, SO₂NH, SO₂NR³, NHSO₂, NR¹, CR¹]*² _, NR¹, or O;

Y is (CHR⁴)_n wherein n = 1-5 or a C3-C7 carbocyclic or aromatic divalent moiety;

A is a bond (absent), C(=0), NH, NR⁵, C(0)NH, C(0)NR⁶, C(S)NH, C(S)NR⁶, NHC(O), NR⁶C(0), NHSO₂, NR⁶S0₂, SO₂NH, SO₂NR⁶, or OP(=0)(OR⁷);

Het is substituted or unsubstituted Ci_-8 alkyl, C₂-8 alkenyl, C₂-8 alkynyl, aryl, fused aryl, heteroaryl, fused heterocycle, a saturated or unsaturated carbocyclic ring or a heterocyclic ring, or a group of the formula -(CH₂)_q-Q where q is 1-4 and Q is an aryl, heteroaryl, cycloalkyl or heterocycle and Q is optionally substituted,

where each Het is optionally substituted and may contain a heteroatom selected from N, O and S in place of a ring or chain carbon atom of Het;

each of R¹, R², R³, R⁴, R⁵, R⁶, and R⁷ is independently selected from H, OH, Halo, NHR, NRR, OR, SR, COOR, C(=0)R, CN, CF₃, OCF₃, N0₂, OC(0)R, S0₃R, PO3R2,

CR(COOR)₂;

each R is independently selected from H, halo, OR^x, SR^X, C0₂R^x, C(0)NR^x ₂, C(=0) R^x, CN, CF₃, OCF3, N0₂, NR^X2, OCOR^x, S0₃R^x, P0₃R^x ₂, and CH(COOR^x)₂;

where each R^x is H or C1-C4 alkyl or C1-C4 haloalkyl, or R can be substituted or unsubstituted Ci_g alkyl, C2-8 alkenyl, C2-8 alkynyl, aryl, fused aryl, heteroaryl, fused heterocycle, a C₃-8 carbocyclic ring or a C₃-8 heterocyclic ring, saturated or unsaturated, wherein each Ci_-8 alkyl, C₃-8 cyclic alkyl, C2-8 alkenyl, C2-8 alkynyl can optionally contain a heteroatom selected from N, O and S in place of a carbon atom; p is 0-3;

each R¹⁰ is H or acyl;

1 2 3 4 5 6 V

and two R, R , R , R , R , R , R , or R on the same or adjacent atoms can optionally be linked together to form a 3-8 membered ring that can contain up to two heteroatoms selected from N, O and S as ring members and which is optionally substituted;

or a pharmaceutically acceptable salt thereof.

[0008] In another aspect, the invention provides a compound of Formula Ila and lib:

A com ound of Formula Ila and lib:

Ila lib

wherein:

W¹ is NR\ O or S;

W² is N or CR²;

Ar is a 5-6 membered aryl or heteroaryl ring that can be substituted or unsubstituted; and

R¹, R², R¹⁰, Z, Y, A, and Het are as defined for Formula la and lb;

or a pharmaceutically acceptable salt thereof.

[0009] In another embodiment the compounds can be of Formula Ilia and Illb

wherein: W¹ is NR\ O or S;

Z is S, SO, S0₂, SO₂NH, SO₂NR³, NHSO₂, NR¹, CR^² NR¹, or O; Y is -CH₂- or -CH₂-CH₂- or -CH(Me)-;

A is a bond or C(=0);

R¹⁰ is H;

R² and Het are as defined for Formula la and lb above;

Ph is an optionally substituted phenyl group;

or a pharmaceutically acceptable salt thereof.

[0010] In a further embodiment, the invention provides a compound of Formula IVa-d:

Wherein:

Het is as defined for Formula la and lb

n is 1-4

R¹, R² and R³ is independently selected from H, OH, Halo, NHR' , NR'R' , OR' , SR', COOR', C(=0)R' , CN, CF₃, OCF₃, N0₂, OC(0)R', S0₃R' , P0₃R'₂,

CR'(COOR')₂

each R' is independently selected from H, halo, OR^x, SR^X, C0₂R^x, C(0)NR^X2, C(=0) R^x, CN, CF₃, OCF₃, N0₂, NR^X ₂, OCOR^x, S0₃R^x, P0₃R^x ₂, and

CH(COOR^x)₂;

where each R^x is H or C1-C4 alkyl or C1-C4 haloalkyl,

or a pharmaceutically acceptable salt thereof. [0011] In another aspect, the invention provides a compound of Formula V:

A compound of Formula Va or Vb:

Formula Va Formula Vb

Wherein:

Z is S, SO, S0₂, Ci^R² NR^JR², or O;

A is a bond, C=0, CONH or NHCO;

Het is as defined for Formula la and lb;

n is 1-4;

R¹, R² and R³ is independently selected from H, OH, Halo, NHR', NR'R',

OR', SR', COOR', C(=0)R', CN, CF₃, OCF₃, N0₂, OC(0)R', S0₃R', P0₃R'₂,

CR'(COOR')₂;

each R' is independently selected from H, halo, OR^x, SR^X, C0₂R^x,

C(0)NR^X2, C(=0) R^x, CN, CF₃, OCF₃, N0₂, NR^X ₂, OCOR^x, S0₃R^x, P0₃R^x ₂, and

CH(COOR^x)₂;

where each R^x is H or C1-C4 alkyl or C1-C4 haloalkyl;

or a pharmaceutically acceptable salt thereof.

[0012] Other aspects of the invention provide pharmaceutical compositions comprising a compound of the invention and methods of using these compounds and compositions for treating proliferative disorders such as cancers.

Brief Description of the Figures

[0013] Figure 1 shows the TCRP (time-dependent cell response profiling) of c-Met addicted H1993 cells in response to the treatment of various heterocyclic compounds.

[0014] Figure 2 shows the TCRP (time-dependent cell response profiling) of c-Met addicted GTL16 cells in response to the treatment of various heterocyclic compounds. [0015] Figure 3 shows the TCRP (time-dependent cell response profiling) of c-Met-non- addicted A549 cells in response to the treatment of various heterocyclic compounds.

[0016] Figure 4 shows the impedance response profile of HGF- mediated migration of A431 cells to the treatment of various heterocyclic compounds

[0017] Figure 5 shows the impedance response profile of HGF-mediated migration of HUVECs (human umbilical vein endothelial cells) to the treatment of various heterocyclic compounds

[0018] Figure 6 shows one of the heterocyclic compounds (compoundl3) inhibited cMet phosphorylation in A549 and GTL16 cell lines.

[0019] Figure 7 shows the inhibitory effects of Compoundl3, compound20, compoundl9 and compound21 on ROS1 phosphorylation in U138MG cells.

Detailed Description

[0020] In one aspect, the invention provides a compound of Formula la and lb:

Formula la Formula lb where in a ring indicates the ring is an aromatic or heteroaromatic ring;

W¹, W² W³ _, W⁴ are each independently absent, N, NH, NR¹, O, S, CH, or CR²,

provided at least one of W¹, W² W³ _, and W⁴ is CH or CR²,

not more than one of them is absent,

and not more than one of them is O or S;

R' is Ci-8 alkyl, C₂-8 alkenyl, C₂-8 alkynyl, aryl, or heteroaryl, wherein each alkyl, alkenyl, alkynyl, aryl and heteroaryl is optionally substituted;

Z is S, SO, S0₂, S0₂NH, S0₂NR³, NHSO₂, NR¹, CR¹]*² _, NR¹, or O;

Y is (CHR⁴)_n wherein n = 1-5 or a C3-C7 carbocyclic or aromatic divalent moiety;

A is a bond (absent), C(=0), NH, NR⁵, C(0)NH, C(0)NR⁶, C(S)NH, C(S)NR⁶, NHC(O), NR⁶C(0), NHSO₂, NR⁶S0₂, S0₂NH, S0₂NR⁶, or OP(=0)(OR⁷);

Het is substituted or unsubstituted Ci_-8 alkyl, C₂-8 alkenyl, C₂-8 alkynyl, aryl, fused aryl, heteroaryl, fused heterocycle, a saturated or unsaturated carbocyclic ring or a heterocyclic ring, or a group of the formula -(CH₂)_q-Q where q is 1-4 and Q is an aryl, heteroaryl, cycloalkyl or heterocycle and Q is optionally substituted,

where each Het is optionally substituted and may contain a heteroatom selected from N, O and S in place of a ring or chain carbon atom of Het;

each of R¹, R², R³, R⁴, R⁵, R⁶, and R⁷ is independently selected from H, OH, Halo, NHR, NRR, OR, SR, COOR, C(=0)R, CN, CF₃, OCF₃, N0₂, OC(0)R, S0₃R, P0₃R₂,

CR(COOR)₂;

each R is independently selected from H, halo, OR^x, SR^X, C0₂R^x, C(0)NR^x ₂, C(=0) R^x, CN, CF₃, OCF₃, N0₂, NR^X2, OCOR^x, S0₃R^x, P0₃R^x ₂, and CH(COOR^x)₂;

where each R^x is H or C1-C4 alkyl or C1-C4 haloalkyl,

or R can be substituted or unsubstituted Ci_-8 alkyl, C₂_8 alkenyl, C₂_8 alkynyl, aryl, fused aryl, heteroaryl, fused heterocycle, a C₃_8 carbocyclic ring or a C₃_8 heterocyclic ring, saturated or unsaturated, wherein each Ci_-8 alkyl, C₃_8 cyclic alkyl, C₂_8 alkenyl, C₂_8 alkynyl can optionally contain a heteroatom selected from N, O and S in place of a carbon atom; p is 0-3;

each R¹⁰ is H or acyl;

1 2 3 4 5 6 V

and two R, R , R , R , R , R , R , or R on the same or adjacent atoms can optionally be linked together to form a 3-8 membered ring that can contain up to two heteroatoms selected from N, O and S as ring members and which is optionally substituted;

or a pharmaceutically acceptable salt thereof.

[0021] In some embodiments of these compounds, W⁴ is CR² and W³ is a bond.

[0022] In some embodiments of any of the foregoing compounds, W¹ is NR¹. Rl for these compounds is often H or a lower alkyl, e.g. , C1-C4 alkyl (Me, Et). In other embodiments, W¹ is O or S.

[0023] In some embodiments of any of the foregoing compounds R¹⁰ is H. In other embodiments, R¹⁰ is an acyl group, typically comprising a lower alkyl or lower haloalkyl {e.g. , up to about 4 carbon atoms).

[0024] In some embodiments of any of the foregoing compounds, Z is S, SO, S0₂, S0₂NH, S0₂NR³, NHS0₂, NR¹, CR^² NR¹, or O.

[0025] In some embodiments of any of the foregoing compounds, Y is (CH₂)i_₄. In many embodiments, Y is CH₂, CH(Me) or CH₂CH₂.

[0026] In some embodiments of any of the foregoing compounds, A is a bond or C(=0). [0027] In some embodiments of any of the foregoing compounds, Het is a 5 or 6 membered aromatic or heteroaromatic ring. In some such embodiments, Het is phenyl or pyridyl, and is optionally substituted with up to three R groups as described above. In some of these embodiments, the R substituents are selected from Me, OMe, SMe, CF₃, CN, and halo.

[0028] In some embodiments of any of the foregoing compounds, Het is phenyl or pyridyl and is optionally substituted with up to three R groups, often 1-2 R groups.

[0029] In some embodiments of any of the compounds of Formula I, W⁴ is CR², wherein R² is H, Cl-4 alkyl, or Cl-4 haloalkyl.

[0030] In certain embodiments of any of the foregoing compounds, R' is optionally substituted phenyl. Typically this phenyl is optionally substituted with up to three substituents groups as described herein. In some of these embodiments, the substituents are selected from Me, OMe, SMe, CF₃, CN, and halo.

[0031] In certain embodiments of any of the foregoing compounds, W² is N.

[0032] In another as ect, the compounds of the invention are of Formula Ila and lib:

Ila lib

wherein:

W¹ is NR\ O or S;

W² is N or CR²;

Ar is a 5-6 membered aryl or heteroaryl ring that can be substituted or unsubstituted; and

R¹, R², R¹⁰, Z, Y, A, and Het are as defined in Formula la and lb above; or a pharmaceutically acceptable salt thereof.

[0033] In the compounds of Formula Ila and lib, any or all of the following further selections may be utilized:

R¹⁰ is H.

W¹ is NR^ or W¹ is O or S.

W² is N or CR².

Z is S, SO, S0₂, SO₂NH, SO₂NR³, NHSO₂, NR¹, C^R² NR¹, or O;

Y is (CH₂)_1-4. A is a bond or C(=0).

Het is a 5 or 6 membered aromatic or heteroaromatic ring; for example, Het is phenyl or pyridyl and is optionally substituted with up to three R groups.

R² is H, Ci-4 alkyl, or C₁-4 haloalkyl.

Ar is optionally substituted phenyl.

[0034] In some embodiments of the compounds of Formula Ila and lib, Het is one of the following

wherein the wavy line across one non-cyclic bond indicates the bond that connects each Het group to A; where in a ring indicates the ring is an aromatic or heteroaromatic ring;

each W and W is independently a bond, C, CH, CH₂, CHR¹², CR¹², N, NH, NR¹², S, O, C=0, or C=S,

where each R is independently selected from H, halo, OR^x, SR^X, C0₂R^x, C(0)NR^x ₂, C(=0) R^x, CN, CF₃, OCF3, N0₂, NR^X2, OCOR^x, S0₃R^x, P0₃R^x ₂, and CH(COOR^x)₂, where each R^x is H or C1-C4 alkyl or C1-C4 haloalkyl,

or R can be substituted or unsubstituted Ci_-8 alkyl, C₂-8 alkenyl, C₂-8 alkynyl;

and each R¹² is H or C1-C4 alkyl or C1-C4 haloalkyl,

or a pharmaceutically acceptable salt thereof.

[0035] In these embodiments, one or more R groups may be selected from: H, halo, OR^x, S(0)_tR^x, C0₂R^x, C(0)NR^X2, C(=0) R^x, CN, CF₃, OCF₃, N0₂, NR^X ₂, OCOR^x, S0₃R^x, P0₃R^x ₂, CH(COOR^x)₂, where each R^x is H or C1-C4 alkyl or C1-C4 haloalkyl, and each t is 0-2.

[0036] In these embodiments, one or more R groups may be selected from: Ci_-8 alkyl, C₃_8 cyclic alkyl, C₂-8 alkenyl, C₂-8 alkynyl, an aryl, heteroaryl, a carbocyclic ring or a heterocyclic ring, each of which may contain a heteroatom selected from N, O and S in place of a carbon atom of the alkyl, alkenyl, alkynyl group.

[0037] In another aspect, the compounds of Formula Ila and lib may be compounds of Formula Ilia and Illb:

wherein:

Z is S, SO, S0₂, SO₂NH, SO₂NR³, NHSO₂, NR¹, CR^² NR¹, or O; Y is -CH₂- or -CH₂-CH₂- or -CH(Me)-;

A is a bond or C(=0);

Het, R¹ and R² are as defined for Formula la or lb above;

R¹⁰ is H;

Ph is an optionally substituted phenyl group;

or a pharmaceutically acceptable salt thereof.

[0038] In the com ounds of Formula Ilia and Illb, Het may be one of the following groups:

wherein the wavy line across one non-cyclic bond indicates the bond that connects each Het group to A; where in a ring indicates the ring is an aromatic or heteroaromatic ring;

each W and W is independently a bond, C, CH, CH₂, CHR¹², CR¹², N, NH, NR¹², S, O,

C=0, or C=S, where each R is independently selected from H, halo, OR^x, SR^X, C0₂R^x, C(0)NR^x ₂, C(=0) R^x, CN, CF₃, OCF₃, N0₂, NR^X2, OCOR^x, S0₃R^x, P0₃R^x ₂, and CH(COOR^x)₂, where each R^x is H or C1-C4 alkyl or C1-C4 haloalkyl,

or R can be substituted or unsubstituted Ci_-8 alkyl, C2-8 alkenyl, C2-8 alkynyl;

and each R¹² is H or C1-C4 alkyl or C1-C4 haloalkyl.

[0039] An embodiment of Formula Ilia and Illb may be where Het is phenyl or pyridyl, it is sometimes optionally substituted with up to three groups selected from C1-C4 alkyl, C1-C4 haloalkyl, halo, OR^x, SR^X, C0₂R^x, C(0)NR^x ₂, C(=0) R^x, CN, CF₃, OCF₃, N0₂, NR^X ₂, OCOR^x, S0₃R^x, P0₃R^X2, and CH(COOR^x)₂, where each R^x is H or C1-C4 alkyl or C1-C4 haloalkyl.

[0040] In a further embodiment, the invention provides a compound of Formula IVa-d:

wherein:

Het is as defined for Formula la and lb

n is 1-4

R¹, R² and R³ is independently selected from H, OH, Halo, NHR' , NR'R' , OR' , SR', COOR', C(=0)R\ CN, CF₃, OCF₃, N0₂, OC(0)R', S0₃R' , P0₃R'₂,

CR'(COOR')₂

each R' is independently selected from H, halo, OR^x, SR^X, C0₂R^x, C(0)NR^X2, C(=0) R^x, CN, CF₃, OCF₃, N0₂, NR^X ₂, OCOR^x, S0₃R^x, P0₃R^x ₂, and

CH(COOR^x)₂;

where each R^x is H or C1-C4 alkyl or C1-C4 haloalkyl, or a pharmaceutically acceptable salt thereof.

[0041] In the compounds of Formula IVa, IVb, IVc or IVd, Het may be one of the following groups:

wherein the wavy line across one non-cyclic bond indicates the bond that connects each Het group to A; where in a ring indicates the ring is an aromatic or heteroaromatic ring;

each W and W is independently a bond, C, CH, CH₂, CHR¹², CR¹², N, NH, NR¹², S, O, C=0, or C=S,

where each R is independently selected from H, halo, OR^x, SR^X, C0₂R^x, C(0)NR^x ₂, C(=0) R^x, CN, CF₃, OCF₃, N0₂, NR^X2, OCOR^x, S0₃R^x, P0₃R^x ₂, and CH(COOR^x)₂, where each R^x is H or C1-C4 alkyl or C1-C4 haloalkyl,

or R can be substituted or unsubstituted Ci_-8 alkyl, C2-8 alkenyl, C2-8 alkynyl;

and each R¹² is H or C1-C4 alkyl or C1-C4 haloalkyl.

[0042] An embodiment of Formula IVa, IVb, IVc or IVd may be where Het is phenyl or pyridyl, it is sometimes optionally substituted with up to three groups selected from C1-C4 alkyl, C1-C4 haloalkyl, halo, OR^x, SR^X, C0₂R^x, C(0)NR^x ₂, C(=0) R^x, CN, CF₃, OCF₃, N0₂, NR^X2, OCOR^x, S0₃R^x, P0₃R^X2, and CH(COOR^x)₂, where each R^x is H or C1-C4 alkyl or C1-C4 haloalkyl.

[0043] In another aspect, the invention provides a compound of Formula Va or Vb:

A compound of Formula Va or Vb:

Wherein:

Z is S, SO, S0₂, Ci^R² NR^JR², or O;

A is a bond, C=0, CONH or NHCO;

Het is as defined for Formula la or lb;

n is 1-4;

R¹, R² and R³ is independently selected from H, OH, Halo, NHR' , NR'R' , OR' , SR', COOR', C(=0)R' , CN, CF₃, OCF₃, N0₂, OC(0)R', S0₃R' , P0₃R'₂,

CR'(COOR')₂;

each R' is independently selected from H, halo, OR^x, SR^X, C0₂R^x, C(0)NR^X2, C(=0) R^x, CN, CF₃, OCF₃, N0₂, NR^X ₂, OCOR^x, S0₃R^x, P0₃R^x ₂, and

CH(COOR^x)₂;

where each R^x is H or C1-C4 alkyl or C1-C4 haloalkyl;

or a pharmaceutically acceptable salt thereof.

[0044] In the compounds of Formula Va or Vb, Het may be one of the following groups:

wherein the wavy line across one non-cyclic bond indicates the bond that connects each Het group to A; where ^ι--' in a ring indicates the ring is an aromatic or heteroaromatic ring;

each W and W is independently a bond, C, CH, CH₂, CHR¹², CR¹², N, NH, NR¹², S, O, C=0, or C=S,

where each R is independently selected from H, halo, OR^x, SR^X, C0₂R^x, C(0)NR^x ₂, C(=0) R^x, CN, CF₃, OCF₃, N0₂, NR^X2, OCOR^x, S0₃R^x, P0₃R^x ₂, and CH(COOR^x)₂, where each R^x is H or C1-C4 alkyl or C1-C4 haloalkyl,

or R can be substituted or unsubstituted Ci_-8 alkyl, C2-8 alkenyl, C2-8 alkynyl;

and each R¹² is H or C1-C4 alkyl or C1-C4 haloalkyl.

[0045] An embodiment of Formula Va or Vb may be where Het is phenyl or pyridyl, it is sometimes optionally substituted with up to three groups selected from C1-C4 alkyl, C1-C4 haloalkyl, halo, OR^x, SR^X, C0₂R^x, C(0)NR^x ₂, C(=0) R^x, CN, CF₃, OCF₃, N0₂, NR^X ₂, OCOR^x, S0₃R^x, P0₃R^x ₂, and CH(COOR^x)₂, where each R^x is H or C1-C4 alkyl or C1-C4 haloalkyl.

[0046] In another aspect, the invention provides a pharmaceutical composition comprising a compound according to any of the foregoing embodiments admixed with at least one pharmaceutically acceptable carrier or excipient. Suitable carriers and excipients are described herein. In some embodiments, this pharmaceutical composition comprises at least one sterile pharmaceutically acceptable carrier or excipient. In some embodiments, the composition comprises at least two pharmaceutically acceptable carriers and/or excipients.

[0047] In another aspect, the invention provides a compound according to any one of the foregoing embodiments for use in therapy. In particular embodiments of interest, the compound is for use in therapy to treat cancer, e.g. , a cancer selected from leukemia, lymphoma, lung cancer, colon cancer, CNS cancer, melanoma, ovarian cancer, renal cancer, prostate cancer, breast cancer, head and neck cancers, and pancreatic cancer.

[0048] In one aspect, the invention provides a method to treat cancer, which comprises administering to a subject in need thereof an effective amount of a compound according to any of the foregoing embodiments, or a pharmaceutical composition comprising one or more of such compounds. In some embodiments, the cancer is selected from leukemia, lymphoma, lung cancer, colon cancer, CNS cancer, melanoma, ovarian cancer, renal cancer, prostate cancer, breast cancer, head and neck cancers, and pancreatic cancer.

[0049] Similarly, the invention provides use of a compound according to any one of the foregoing embodiments for the manufacture of a medicament. In some embodiments, the medicament is one for treating cancer, and in some embodiments the cancer is selected from leukemia, lymphoma, lung cancer, colon cancer, CNS cancer, melanoma, ovarian cancer, renal cancer, prostate cancer, breast cancer, head and neck cancers, and pancreatic cancer.

[0050] In the above formula I - V, "Het" may be mono-/di-/tri-substituted or unsubstituted benzene, pyridine, pyridazine, pyrimidine, pyrazine, triazine, oxazole, isoxazole, thiazole, isothiazole, oxadiazole, triazole, thiadiazole, pyrazole, imidazole, benzoxazole, pyrrole, furan, thiophene, indolizine, indole, isoindole, indoline, benzofuran, benzothiophene, indazole, benzimidazole, benzthiazole, purine, quinoxaline, quinoline, isoquinoline, cinnoline, phthalazine, quinazoline, naphthyridine, pteridine, acridine, phenazine, phenothiazine, indene, naphthalene or benzoxadiazole. Illustrated examples of "Het", include but are not limited to the fo

[0051] It will be understood that the selections of W¹, W², W³, W⁴, W and Z for any of the above cyclic groups would of course be made by a person of ordinary skill in the art in accordance with well known limitations associated with valence rules and stability. These atoms or bonds would thus be selected to provide only a stable ring or ring system consistent with well-known bonding and valence principles; thus these rings would not be constructed with -O-O- or -S-S- linkages in the rings, for example, or with inappropriate numbers of O or N or S atoms causing instability in aqueous media. Typically each ring of these compounds will be a 5-6 atom ring, whether aromatic or not, and will contain at least one carbon atom, no more than one O or S atom (except for a dioxane ring having two O atoms), and up to four N atoms as ring members. Non-aromatic rings will typically contain no more than two heteroatoms in place of ring carbon atoms, while aromatic rings containing 3-4 heteroatoms— especially N atoms— such as triazines, triazoles, tetrazines and tetrazoles, are included. Likewise, other groups such as R would be selected to avoid compounds generally considered to be too reactive for use as pharmaceuticals, e.g. , R would not be Halo in groups such as NHR, NRR, OR, SR, or COOR. [0052] Suitable dosages, routes of administration, and administration protocols can be selected by a skilled person based on the information herein.

General Definitions:

[0053] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of ordinary skill in the art to which this invention belongs. All patents, applications, published applications and other publications referred to herein are incorporated by reference in their entireties. If a definition set forth in this section is contrary to or otherwise inconsistent with a definition set forth in a patent, application, or other publication that is herein incorporated by reference, the definition set forth in this section prevails over the definition incorporated herein by reference.

[0054] As used herein, "a" or "an" means "at least one" or "one or more".

[0055] The term "alkyl" as used herein refers to saturated hydrocarbon groups in a straight, branched, or cyclic configuration or any combination thereof, and particularly contemplated alkyl groups include those having ten or less carbon atoms, especially 1-6 carbon atoms and lower alkyl groups having 1-4 carbon atoms. Exemplary alkyl groups are methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tertiary butyl, pentyl, isopentyl, hexyl, cyclopropylmethyl, etc.

[0056] Alkyl groups can be unsubstituted, or they can be substituted to the extent that such substitution makes sense chemically. Typical substituents include, but are not limited to, halo, =0, =N-CN, =N-OR^a, =NR^a, -OR^a, -NR^a ₂, -SR^a, -S0₂R^a, -S0₂NR^a ₂, -NR^aS0₂R^a, -NR^aCONR^a ₂, -NR^aCOOR^a, -NR^aCOR^a, -CN, -COOR^a, -CONR^a ₂, -OOCR^a, -COR^a, and -N0₂, wherein each R^a is independently H, C1-C8 alkyl, C2-C8 heteroalkyl, C3-C8 heterocyclyl,

C4-C10 heterocyclyclalkyl, C1-C8 acyl, C2-C8 heteroacyl, C2-C8 alkenyl,

C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl, C6-C10 aryl, or C5-C10 heteroaryl, and each R^a is optionally substituted with halo, =0, =N-CN, =N-OR^b, =NR^b, OR^b, NR^b ₂, SR^b, S0₂R^b, S0₂NR^b ₂, NR^bS0₂R^b, NR^bCONR^b ₂, NR^bCOOR^b, NR^bCOR^b, CN, COOR^b, CONR^b ₂, OOCR^b, COR^b, and N0₂, wherein each R^b is independently H, C1-C8 alkyl, C2-C8 heteroalkyl, C3-C8 heterocyclyl, C4-C10 heterocyclyclalkyl, C1-C8 acyl, C2-C8 heteroacyl, C6-C10 aryl or C5-C10 heteroaryl. Alkyl, alkenyl and alkynyl groups can also be substituted by C1-C8 acyl, C2-C8 heteroacyl, C6-C10 aryl or C5-C10 heteroaryl, each of which can be substituted by the substituents that are appropriate for the particular group. Where a substituent group contains two R^a or R^b groups on the same or adjacent atoms (e.g., -NR^b2, or -NR^b-C(0) R^b), the two R^a or R^b groups can optionally be taken together with the atoms in the substituent group to which the are attached to form a ring having 5-8 ring members, which can be substituted as allowed for the R^a or R^b itself, and can contain an additional heteroatom (N, O or S) as a ring member. [0057] The term "alkenyl" as used herein refers to an alkyl as defined above having at least two carbon atoms and at least one carbon-carbon double bond. Thus, particularly contemplated alkenyl groups include straight, branched, or cyclic alkenyl groups having two to ten carbon atoms (e.g., ethenyl, propenyl, butenyl, pentenyl, etc.) or 5-10 atoms for cyclic alkenyl groups. Alkenyl groups are optionally substituted by groups suitable for alkyl groups as set forth herein.

[0058] Similarly, the term "alkynyl" as used herein refers to an alkyl or alkenyl as defined above and having at least two (preferably three) carbon atoms and at least one carbon-carbon triple bond. Especially contemplated alkynyls include straight, branched, or cyclic alkynes having two to ten total carbon atoms (e.g. , ethynyl, propynyl, butynyl, cyclopropylethynyl, etc.). Alkynyl groups are optionally substituted by groups suitable for alkyl groups as set forth herein.

[0059] The term "cycloalkyl" as used herein refers to a cyclic alkane (i.e. , in which a chain of carbon atoms of a hydrocarbon forms a ring), preferably including three to eight carbon atoms. Thus, exemplary cycloalkanes include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Cycloalkyls also include one or two double bonds, which form the "cycloalkenyl" groups. Cycloalkyl groups are optionally substituted by groups suitable for alkyl groups as set forth herein.

[0060] The term "aryl" or "aromatic moiety" as used herein refers to an aromatic ring system, which may further include one or more non-carbon atoms. These are typically 5-6 membered isolated rings, or 8-10 membered bicyclic groups, and can be substituted. Thus, contemplated aryl groups include (e.g., phenyl, naphthyl, etc.) and pyridyl. Further

contemplated aryl groups may be fused (i.e., covalently bound with 2 atoms on the first aromatic ring) with one or two 5- or 6-membered aryl or heterocyclic group, and are thus termed "fused aryl" or "fused aromatic".

[0061] Aromatic groups containing one or more heteroatoms (typically N, O or S) as ring members can be referred to as heteroaryl or heteroaromatic groups. Typical heteroaromatic groups include monocyclic C5-C6 aromatic groups such as pyridyl, pyrimidyl, pyrazinyl, thienyl, furanyl, pyrrolyl, pyrazolyl, thiazolyl, oxazolyl, isothiazolyl, isoxazolyl, and imidazolyl and the fused bicyclic moieties formed by fusing one of these monocyclic groups with a phenyl ring or with any of the heteroaromatic monocyclic groups to form a C8-C10 bicyclic group such as indolyl, benzimidazolyl, indazolyl, benzotriazolyl, isoquinolyl, quinolyl, benzothiazolyl, benzofuranyl, pyrazolopyridyl, pyrazolopyrimidyl, quinazolinyl, quinoxalinyl, cinnolinyl, and the like. Any monocyclic or fused ring bicyclic system which has the characteristics of aromaticity in terms of electron distribution throughout the ring system is included in this definition. It also includes bicyclic groups where at least the ring which is directly attached to the remainder of the molecule has the characteristics of aromaticity. Typically, the ring systems contain 5- 12 ring member atoms.

[0062] As also used herein, the terms "heterocycle", "cycloheteroalkyl", and "heterocyclic moieties" are used interchangeably herein and refer to any compound in which a plurality of atoms form a ring via a plurality of covalent bonds, wherein the ring includes at least one atom other than a carbon atom as a ring member. Particularly contemplated heterocyclic rings include 5- and 6-membered rings with nitrogen, sulfur, or oxygen as the non-carbon atom (e.g. , imidazole, pyrrole, triazole, dihydropyrimidine, indole, pyridine, thiazole, tetrazole etc.).

Typically these rings contain 0-1 oxygen or sulfur atoms, at least one and typically 2-3 carbon atoms, and up to four nitrogen atoms as ring members. Further contemplated heterocycles may be fused (i.e., covalently bound with two atoms on the first heterocyclic ring) to one or two carbocyclic rings or heterocycles, and are thus termed "fused heterocycle" or "fused

heterocyclic ring" or "fused heterocyclic moieties" as used herein. Where the ring is aromatic, these can be referred to herein as 'heteroaryl' or heteroaromatic groups.

[0063] Heterocyclic groups that are not aromatic can be substituted with groups suitable for alkyl group substituents, as set forth above.

[0064] Aryl and heteroaryl groups can be substituted where permitted. Suitable substituents include, but are not limited to, halo, -OR^a, -NR^a ₂, -SR^a, -S0₂R^a, -S0₂NR^a ₂, -NR^aS0₂R^a, -NR^aCONR^a ₂, -NR^aCOOR^a, -NR^aCOR^a, -CN, -COOR^a, -CONR^a ₂, -OOCR^a, -COR^a, and -N0₂, wherein each R^a is independently H, Ci-C₈ alkyl, C2-C8 heteroalkyl, C3-C8 heterocyclyl, C4-C10 heterocyclyclalkyl, C1-C8 acyl, C2-C8 heteroacyl, C2-C8 alkenyl, C2-C8

heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl, C6-C10 aryl, or C5-C10 heteroaryl, and each R^a is optionally substituted with halo, =0, =N-CN, =N-OR^b, =NR^b, OR^b, NR^b ₂, SR^b, S0₂R^b, S0₂NR^b ₂, NR^bS0₂R^b, NR^bCONR^b ₂, NR^bCOOR^b, NR^bCOR^b, CN, COOR^b, CONR^b ₂, OOCR^b, COR^b, and N0₂, wherein each R^b is independently H, C1-C8 alkyl, C2-C8 heteroalkyl, C3-C8 heterocyclyl, C4-C10 heterocyclyclalkyl, C1-C8 acyl, C2-C8 heteroacyl, C6-C10 aryl or C5-C10 heteroaryl. Alkyl, alkenyl and alkynyl groups can also be substituted by C1-C8 acyl, C2-C8 heteroacyl, C6-C10 aryl or C5-C10 heteroaryl, each of which can be substituted by the substituents that are appropriate for the particular group. Where a substituent group contains two R^a or R^b groups on the same or adjacent atoms (e.g., -NR^b2, or -NR^b-C(0) R^b), the two R^a or R^b groups can optionally be taken together with the atoms in the substituent group to which the are attached to

a b

form a ring having 5-8 ring members, which can be substituted as allowed for the R or R itself, and can contain an additional heteroatom (N, O or S) as a ring member. [0065] As also used herein, the terms "imidazopyridine" or "imidazopyrimidine" or "thiazopyridine" or "thiazopyrimidine" herein refer to any compound in which the two designated heterocyclic rings are fused by any two adjacent atoms on the two heterocyclic rings.

[0066] The term "alkoxy" as used herein refers to a hydrocarbon group connected through an oxygen atom, e.g., -O-Hc, wherein the hydrocarbon portion He may have any number of carbon atoms, typically 1-10 carbon atoms, may further include a double or triple bond and may include one or two oxygen, sulfur or nitrogen atoms in the alkyl chains, and can be substituted with aryl, heteroaryl, cycloalkyl, and/or heterocyclyl groups. For example, suitable alkoxy groups include methoxy, ethoxy, propyloxy, isopropoxy, methoxyethoxy, benzyloxy, allyloxy, and the like. Similarly, the term "alkylthio" refers to alkylsulfides of the general formula -S- Hc, wherein the hydrocarbon portion He is as described for alkoxy groups. For example, contemplated alkylthio groups include methylthio, ethylthio, isopropylthio, methoxyethylthio, benzylthio, allylthio, and the like.

[0067] The term 'amino' as used herein refers to the group -N¾. The term "alkylamino" refers to amino groups where one or both hydrogen atoms are replaced by a hydrocarbon group He as described above, wherein the amino nitrogen "N" can be substituted by one or two He groups as set forth for alkoxy groups described above. Exemplary alkylamino groups include methylamino, dimethylamino, ethylamino, diethylamino, etc.

[0068] The term 'acyl' as used herein refers to a group of the formula -C(=0)-D, where D represents an alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, or heterocycle as described above. Typical examples are groups wherein D is a C1-C10 alkyl, C2-C10 alkenyl or alkynyl, or phenyl, each of which is optionally substituted. In some embodiments, D can be H, Me, Et, isopropyl, propyl, butyl, C1-C4 alkyl substituted with -OH, -OMe, or N¾, phenyl, halophenyl, alkylphenyl, and the like.

[0069] The term "aryloxy" as used herein refers to an aryl group connecting to an oxygen atom, wherein the aryl group may be further substituted. For example suitable aryloxy groups include phenyloxy, etc. Similarly, the term "arylthio" as used herein refers to an aryl group connecting to a sulfur atom, wherein the aryl group may be further substituted. For example suitable arylthio groups include phenylthio, etc.

[0070] The hydrocarbon portion of each alkoxy, alkylthio, alkylamino, and aryloxy, etc., can be substituted as appropriate for the relevant hydrocarbon moiety.

[0071] The term "halogen" as used herein refers to fluorine, chlorine, bromine and iodine. Where present as a substituent group, halogen or halo typically refers to F or CI or Br, more typically F or CI. [0072] It should further be recognized that all of the above-defined groups may further be substituted with one or more substituents, which may in turn be substituted with hydroxy, amino, cyano, C1-C4 alkyl, halo, or C1-C4 haloalkyl. For example, a hydrogen atom in an alkyl or aryl can be replaced by an amino, halo or Ci-₄ haloalkyl or alkyl group.

[0073] The term "substituted" as used herein refers to a replacement of a hydrogen atom of the unsubstituted group with a functional group, and particularly contemplated functional groups include nucleophilic groups (e.g., -NH₂, -OH, -SH, -CN, etc.), electrophilic groups (e.g. , C(0)OR, C(X)OH, etc.), polar groups (e.g. , -OH), non-polar groups (e.g. , heterocycle, aryl, alkyl, alkenyl, alkynyl, etc.), ionic groups (e.g. , -NH₃ ^"1"), and halogens (e.g. , -F, -CI), NHCOR, NHCONH₂, OCH₂COOH, OCH₂CONH₂, OCH₂CONHR, NHCH₂COOH, NHCH₂CONH₂, NHSO₂R, OCH₂-heterocycles, PO₃H, SO₃H, amino acids, and all chemically reasonable combinations thereof. Moreover, the term "substituted" also includes multiple degrees of substitution, and where multiple substituents are disclosed or claimed, the substituted compound can be independently substituted by one or more of the disclosed or claimed substituent moieties.

[0074] Any formula given herein is intended to represent compounds having structures depicted by the structural formula as well as certain variations or forms. In particular, compounds of any formula given herein may have asymmetric centers and therefore exist in different enantiomeric forms. All optical isomers and stereoisomers of the compounds of the general formula, and mixtures thereof, are considered within the scope of the formula. Thus, any formula given herein is intended to represent a racemate, one or more enantiomeric forms, one or more diastereomeric forms, one or more atropisomeric forms, and mixtures thereof. Furthermore, certain structures may exist as geometric isomers (i.e., cis and trans isomers), as tautomers, or as atropisomers. Additionally, any formula given herein is intended to refer also to any one of hydrates, solvates, and polymorphs of such compounds, and mixtures thereof, even if such forms are not listed explicitly. In some embodiments, the solvent is water and the solvates are hydrates.

[0075] Any formula given herein is also intended to represent unlabeled forms as well as isotopically labeled forms of the compounds. Isotopically labeled compounds have structures depicted by the formulas given herein except that one or more atoms are replaced by an atom having a selected atomic mass or mass number. Examples of isotopes that can be incorporated into compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine, chlorine, and iodine, such as ²H, ³H, ⁿC, ¹³C, ¹⁴C, ¹⁵N, ¹⁸0, ¹⁷0, ³¹P, ³²P, ³⁵S, ¹⁸F, ³⁶C1, and ¹²⁵I, respectively. Such isotopically labelled compounds are useful in metabolic studies (preferably with ¹⁴C), reaction kinetic studies (with, for example ²H or ³H), detection or imaging techniques [such as positron emission tomography (PET) or single-photon emission computed tomography (SPECT)] including drug or substrate tissue distribution assays, or in radioactive treatment of patients. In particular, an ¹⁸F or ¹¹ C labeled compound may be particularly preferred for PET or SPECT studies. Further, substitution with heavier isotopes such as deuterium (i.e., ²H) may afford certain therapeutic advantages resulting from greater metabolic stability, for example increased in vivo half-life or reduced dosage requirements. Isotopically labeled compounds of this invention and prodrugs thereof can generally be prepared by carrying out the procedures disclosed in the schemes or in the examples and preparations described below by substituting a readily available isotopically labeled reagent for a non-isotopically labeled reagent.

[0076] The nomenclature "Ci__j" or "Ci-Cj" with j > i, when applied herein to a class of substituents, is meant to refer to embodiments of this invention for which each and every one of the number of carbon members, from i to j including i and j, is independently realized. By way of example, the term Ci_3 or C1-C3 refers independently to embodiments that have one carbon member (Ci or CI), embodiments that have two carbon members (C₂ or C2), and embodiments that have three carbon members (C₃ or C3).

[0077] Any disubstituent referred to herein is meant to encompass the various attachment possibilities when more than one of such possibilities are allowed. For example, reference to disubstituent -A-B-, where A≠ B, refers herein to such disubstituent with A attached to a first substituted member and B attached to a second substituted member, and it also refers to such disubstituent with A attached to the second substituted member and B attached to the first substituted member.

[0078] The invention also includes pharmaceutically acceptable salts of the compounds represented by Formula I-V, preferably of those described above and of the specific compounds exemplified herein, and pharmaceutical compositions comprising such salts, and methods of using such salts.

[0079] A "pharmaceutically acceptable salt" is intended to mean a salt of a free acid or base of a compound represented herein that is non-toxic, biologically tolerable, or otherwise biologically suitable for administration to the subject. See, generally, S.M. Berge, et al., "Pharmaceutical Salts," J. Pharm. Sci., 1977, 66, 1-19. Preferred pharmaceutically acceptable salts are those that are pharmacologically effective and suitable for contact with the tissues of subjects without undue toxicity, irritation, or allergic response. A compound described herein may possess a sufficiently acidic group, a sufficiently basic group, both types of functional groups, or more than one of each type, and accordingly react with a number of inorganic or organic bases, and inorganic and organic acids, to form a pharmaceutically acceptable salt.

[0080] Examples of pharmaceutically acceptable salts include sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, phosphates, monohydrogen-phosphates, dihydrogenphosphates, metaphosphates, pyrophosphates, chlorides, bromides, iodides, acetates, propionates, decanoates, caprylates, acrylates, formates, isobutyrates, caproates, heptanoates, propiolates, oxalates, malonates, succinates, suberates, sebacates, fumarates, maleates, butyne-l,4-dioates, hexyne-l,6-dioates, benzoates, chlorobenzoates, methylbenzoates, dinitrobenzoates, hydroxybenzoates, methoxybenzoates, phthalates, sulfonates, methylsulfonates,

propylsulfonates, besylates, xylenesulfonates, naphthalene- 1- sulfonates, naphthalene-2- sulfonates, phenylacetates, phenylpropionates, phenylbutyrates, citrates, lactates, γ- hydroxybutyrates, glycolates, tartrates, and mandelates.

[0081] For a compound of Formula I-V that contains a basic nitrogen, a pharmaceutically acceptable salt may be prepared by any suitable method available in the art, for example, treatment of the free base with an inorganic acid, such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, nitric acid, boric acid, phosphoric acid, and the like, or with an organic acid, such as acetic acid, phenylacetic acid, propionic acid, stearic acid, lactic acid, ascorbic acid, maleic acid, hydroxymaleic acid, isethionic acid, succinic acid, valeric acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, oleic acid, palmitic acid, lauric acid, a pyranosidyl acid, such as glucuronic acid or galacturonic acid, an alpha-hydroxy acid, such as mandelic acid, citric acid, or tartaric acid, an amino acid, such as aspartic acid or glutamic acid, an aromatic acid, such as benzoic acid, 2-acetoxybenzoic acid, naphthoic acid, or cinnamic acid, a sulfonic acid, such as laurylsulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, or ethanesulfonic acid, or any compatible mixture of acids such as those given as examples herein, and any other acid and mixture thereof that are regarded as equivalents or acceptable substitutes in light of the ordinary level of skill in this technology.

[0082] The invention also relates to pharmaceutically acceptable prodrugs of the compounds of Formula I-V, and treatment methods employing such pharmaceutically acceptable prodrugs. The term "prodrug" means a precursor of a designated compound that, following administration to a subject, yields the compound in vivo via a chemical or physiological process such as solvolysis or enzymatic cleavage, or under physiological conditions (e.g., a prodrug on being brought to physiological pH is converted to the compound of Formula I- V). A "pharmaceutically acceptable prodrug" is a prodrug that is non-toxic, biologically tolerable, and otherwise biologically suitable for administration to the subject. Illustrative procedures for the selection and preparation of suitable prodrug derivatives are described, for example, in "Design of Prodrugs", ed. H. Bundgaard, Elsevier, 1985.

[0083] The present invention also relates to pharmaceutically active metabolites of compounds of Formula I-V, and uses of such metabolites in the methods of the invention. A "pharmaceutically active metabolite" means a pharmacologically active product of metabolism in the body of a compound of Formulas I-V or salt thereof. Prodrugs and active metabolites of a compound may be determined using routine techniques known or available in the art. See, e.g., Bertolini et al., /. Med. Chem. 1997, 40, 2011-2016; Shan et al., /. Pharm. Sci. 1997, 86 (7), 765-767; Bagshawe, Drug Dev. Res. 1995, 34, 220-230; Bodor, Adv. Drug Res. 1984, 13, 255- 331; Bundgaard, Design of Prodrugs (Elsevier Press, 1985); and Larsen, Design and

Application of Prodrugs, Drug Design and Development (Krogsgaard-Larsen et al., eds., Harwood Academic Publishers, 1991).

Formulations

[0084] Any suitable formulation of the compounds described herein can be prepared. See generally, Remington's Pharmaceutical Sciences, (2000) Hoover, J. E. editor, 20 th edition, Lippincott Williams and Wilkins Publishing Company, Easton, Pa., pages 780-857. A formulation is selected to be suitable for an appropriate route of administration. Some routes of administration are oral, parenteral, by inhalation, topical, rectal, nasal, buccal, vaginal, via an implanted reservoir, or other drug administration methods. In cases where compounds are sufficiently basic or acidic to form stable nontoxic acid or base salts, administration of the compounds as salts may be appropriate. Examples of pharmaceutically acceptable salts are organic acid addition salts formed with acids that form a physiological acceptable anion, for example, tosylate, methanesulfonate, acetate, citrate, malonate, tartarate, succinate, benzoate, ascorbate, a-ketoglutarate, and a-glycerophosphate. Suitable inorganic salts may also be formed, including hydrochloride, sulfate, nitrate, bicarbonate, and carbonate salts.

Pharmaceutically acceptable salts are obtained using standard procedures well known in the art, for example, by a sufficiently basic compound such as an amine with a suitable acid, affording a physiologically acceptable anion. Alkali metal (e.g. , sodium, potassium or lithium) or alkaline earth metal (e.g. , calcium) salts of carboxylic acids also are made.

[0085] Where contemplated compounds are administered in a pharmacological composition, it is contemplated that the compounds can be formulated in admixture with a pharmaceutically acceptable excipient and/or carrier. For example, contemplated compounds can be administered orally as neutral compounds or as pharmaceutically acceptable salts, or intravenously in a physiological saline solution. Conventional buffers such as phosphates, bicarbonates or citrates can be used for this purpose. Of course, one of ordinary skill in the art may modify the formulations within the teachings of the specification to provide numerous formulations for a particular route of administration. In particular, contemplated compounds may be modified to render them more soluble in water or other vehicle, which for example, may be easily accomplished with minor modifications (salt formulation, esterification, etc.) that are well within the ordinary skill in the art. It is also well within the ordinary skill of the art to modify the route of administration and dosage regimen of a particular compound in order to manage the pharmacokinetics of the present compounds for maximum beneficial effect in a patient.

[0086] The compounds having formula I-V as described herein are generally soluble in organic solvents such as chloroform, dichloromethane, ethyl acetate, ethanol, methanol, isopropanol, acetonitrile, glycerol, N,N-dimethylformamide, N,N-dimetheylaceatmide, dimethylsulfoxide, etc. In one embodiment, the present invention provides formulations prepared by mixing a compound having formula I-V with a pharmaceutically acceptable carrier. In one aspect, the formulation may be prepared using a method comprising: a) dissolving a described compound in a water-soluble organic solvent, a non-ionic solvent, a water-soluble lipid, a cyclodextrin, a vitamin such as tocopherol, a fatty acid, a fatty acid ester, a

phospholipid, or a combination thereof, to provide a solution; and b) adding saline or a buffer containing 1-10% carbohydrate solution. In one example, the carbohydrate comprises dextrose. The pharmaceutical compositions obtained using the present methods are stable and useful for animal and clinical applications.

[0087] Illustrative examples of water soluble organic solvents for use in the present methods include and are not limited to polyethylene glycol (PEG), alcohols, acetonitrile, N-methyl-2- pyrrolidone, N,N-dimethylformamide, N,N-dimethylacetamide, dimethyl sulfoxide, or a combination thereof. Examples of alcohols include but are not limited to methanol, ethanol, isopropanol, glycerol, or propylene glycol.

[0088] Illustrative examples of water soluble non-ionic surfactants for use in the present methods include and are not limited to CREMOPHOR^® EL, polyethylene glycol modified CREMOPHOR^® (polyoxyethyleneglyceroltriricinoleat 35), hydrogenated CREMOPHOR^® RH40, hydrogenated CREMOPHOR^® RH60, PEG-succinate, polysorbate 20, polysorbate 80, SOLUTOL^® HS (polyethylene glycol 660 12-hydroxystearate), sorbitan monooleate, poloxamer, LABRAFIL^® (ethoxylated persic oil), LABRASOL^® (capryl-caproyl macrogol-8- glyceride), GELUCIRE^® (glycerol ester), SOFTIGEN^® (PEG 6 caprylic glyceride), glycerin, glycol-polysorbate, or a combination thereof. [0089] Illustrative examples of water soluble lipids for use in the present methods include but are not limited to vegetable oils, triglycerides, plant oils, or a combination thereof.

Examples of lipid oils include but are not limited to castor oil, polyoxyl castor oil, corn oil, olive oil, cottonseed oil, peanut oil, peppermint oil, safflower oil, sesame oil, soybean oil, hydrogenated vegetable oil, hydrogenated soybean oil, a triglyceride of coconut oil, palm seed oil, and hydrogenated forms thereof, or a combination thereof.

[0090] Illustrative examples of fatty acids and fatty acid esters for use in the present methods include but are not limited to oleic acid, monoglycerides, diglycerides, a mono- or di- fatty acid ester of PEG, or a combination thereof.

[0091] Illustrative examples of cyclodextrins for use in the present methods include but are not limited to alpha-cyclodextrin, beta-cyclodextrin, hydroxypropyl-beta-cyclodextrin, or sulfobutyl ether-beta-cyclodextrin.

[0092] Illustrative examples of phospholipids for use in the present methods include but are not limited to soy phosphatidylcholine, or distearoyl phosphatidylglycerol, and hydrogenated forms thereof, or a combination thereof.

[0093] One of ordinary skill in the art may modify the formulations within the teachings of the specification to provide numerous formulations for a particular route of administration. In particular, the compounds may be modified to render them more soluble in water or other vehicle. It is also well within the ordinary skill of the art to modify the route of administration and dosage regimen of a particular compound in order to manage the pharmacokinetics of the present compounds for maximum beneficial effect in a patient.

Methods of Using the Invented Compounds and Pharmaceutical Compositions thereof

[0094] The present invention also provides pharmaceutical compositions for the treatment of a cell proliferative disorder, comprising any compound having formula I-V,

[0095] To practice the method of the present invention, compounds having formula I-V and pharmaceutical compositions thereof may be administered orally, parenterally, by inhalation, topically, rectally, nasally, buccally, vaginally, via an implanted reservoir, or other drug administration methods. The term "parenteral" as used herein includes subcutaneous, intracutaneous, intravenous, intramuscular, intraarticular, intraarterial, intrasynovial, intrasternal, intrathecal, intralesional and intracranial injection or infusion techniques.

[0096] A sterile injectable composition, such as a sterile injectable aqueous or oleaginous suspension, may be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent. Among the acceptable vehicles and solvents that may be employed include mannitol, water, Ringer' s solution and isotonic sodium chloride solution. Suitable carriers and other pharmaceutical composition components are typically sterile.

[0097] In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium (e.g. , synthetic mono- or diglycerides). Fatty acids, such as oleic acid and its glyceride derivatives, are useful in the preparation of injectables, as are pharmaceutically acceptable oils, such as olive oil or castor oil, especially in their polyoxyethylated versions. These oil solutions or suspensions can also contain a long-chain alcohol diluent or dispersant, or carboxymethyl cellulose or similar dispersing agents. Various emulsifying agents or bioavailability enhancers which are commonly used in the manufacture of pharmaceutically acceptable solid, liquid, or other dosage forms can also be used for the purpose of formulation.

[0098] A composition for oral administration may be any orally acceptable dosage form including, but not limited to, tablets, capsules, emulsions and aqueous suspensions, dispersions and solutions. In the case of tablets for oral use, commonly used carriers include lactose and corn starch. Lubricating agents, such as magnesium stearate, can also be added. For oral administration in a capsule form, useful diluents include lactose and dried corn starch. When aqueous suspensions or emulsions are administered orally, the active ingredient can be suspended or dissolved in an oily phase combined with emulsifying or suspending agents. If needed, certain sweetening, flavoring, or coloring agents can be added. A nasal aerosol or inhalation compositions can be prepared according to techniques well-known in the art of pharmaceutical formulation and can be prepared as solutions in, for example saline, employing suitable preservatives (for example, benzyl alcohol), absorption promoters to enhance bioavailability, and/or other solubilizing or dispersing agents known in the art.

[0099] In addition, the compounds having formula I-V may be administered alone or in combination with other anticancer agents for the treatment of various cancers or conditions. Combination therapies according to the present invention comprise the administration of at least one compound of the present invention or a functional derivative thereof and at least one other pharmaceutically active ingredient. The active ingredient(s) and pharmaceutically active agents may be administered separately or together. The amounts of the active ingredient(s) and pharmaceutically active agent(s) and the relative timings of administration will be selected in order to achieve the desired combined therapeutic effect. [0100] A compound of formula I-V can besides or in addition be administered especially for tumor therapy in combination with chemotherapy, radiotherapy, immunotherapy, surgical intervention, or a combination of these. Long-term therapy and adjuvant therapy in

combination with the treatment strategies, as described above. Other possible treatments are therapy to maintain the patient's status after tumor regression, or even chemopreventive therapy, for example in patients at risk.

[0101] The compounds and compositions described herein can be administered to a subject in need of treatment for a cell proliferation disorder such as cancer, particularly cancers selected from leukemia, lymphoma, lung cancer, colon cancer, CNS cancer, melanoma, ovarian cancer, renal cancer, prostate cancer, breast cancer, head and neck cancers, and pancreatic cancer. The subject is typically a mammal diagnosed as being in need of treatment for one or more of such proliferative disorders, and frequently the subject is a human. The methods comprise administering an effective amount of at least one compound of the invention; optionally the compound may be administered in combination with one or more additional therapeutic agents, particularly therapeutic agents known to be useful for treating the cancer or proliferative disorder afflicting the particular subject.

[0102] The term "treat" or "treating" as used herein is intended to refer to administration of a compound of the invention to a subject for the purpose of creating a therapeutic benefit to a subject suffering from or diagnosed with cancer or cell proliferation disease, disorder or condition. Treating includes preventing, reversing, ameliorating, alleviating, inhibiting the progress of, or lessening the severity of, a cancer or cell proliferation disorder or one or more symptoms of that disorder, as well as the delay of progression of a disease, disorder or condition. The term "subject" refers to a mammalian patient in need of such treatment, such as a human.

[0103] In the inhibitory methods of the invention, an "effective amount" means an amount sufficient to reduce, slow the progression of, or reverse cancer or cell proliferation disorder. Measuring the amount of cell proliferation may be performed by routine analytical methods such as those described below. Such modulation is useful in a variety of settings, including in vitro assays. In such methods, the cell is preferably a GTL-16, H1993 or A431 cells.

[0104] In treatment methods according to the invention, an "effective amount" means an amount or dose sufficient to generally bring about the desired therapeutic benefit in subjects needing such treatment. Effective amounts or doses of the compounds of the invention may be ascertained by routine methods, such as modeling, dose escalation, or clinical trials, taking into account routine factors, e.g., the mode or route of administration or drug delivery, the pharmacokinetics of the agent, the severity and course of the infection, the subject's health status, condition, and weight, and the judgment of the treating physician.

[0105] Once improvement of the patient's disease has occurred, the dose may be adjusted for preventative or maintenance treatment. For example, the dosage or the frequency of administration, or both, may be reduced as a function of the symptoms, to a level at which the desired therapeutic or prophylactic effect is maintained. Of course, if symptoms have been alleviated to an appropriate level, treatment may cease. Patients may, however, require intermittent treatment on a long-term basis upon any recurrence of symptoms. Patients may also require chronic treatment on a long-term basis.

[0106] The term "combination" refers to either a fixed combination in one dosage unit form, or a kit of parts for the combined administration where a compound of the formula (I) and a combination partner (e.g. an other drug as explained below, also referred to as "therapeutic agent" or "co-agent") may be administered independently at the same time or separately within time intervals, especially where these time intervals allow that the combination partners show a cooperative, e.g. synergistic effect. The terms "co-administration" or "combined administration" or the like as utilized herein are meant to encompass administration of the selected combination partner to a single subject in need thereof (e.g. a patient), and are intended to include treatment regimens in which the agents are not necessarily administered by the same route of

administration or at the same time. The term "pharmaceutical combination" as used herein means a product that results from the mixing or combining of more than one active ingredient and includes both fixed and non-fixed combinations of the active ingredients. The term "fixed combination" means that the active ingredients, e.g. a compound of formula (I) and a combination partner, are both administered to a patient simultaneously in the form of a single entity or dosage. The term "non-fixed combination" means that the active ingredients, e.g. a compound of formula (I) and a combination partner, are both administered to a patient as separate entities either simultaneously, concurrently or sequentially with no specific time limits, wherein such administration provides therapeutically effective levels of the two compounds in the body of the patient. The latter also applies to cocktail therapy, e.g. the administration of three or more active ingredients.

Drug Combinations

[0107] The inventive compounds described herein may be used in pharmaceutical compositions or methods in combination with one or more additional active ingredients in the treatment of cancer, cell proliferative disorders or inflammatory conditions. For example, additional active ingredients are those that are known or discovered to be effective in treating cancer, cell proliferative disorders or inflammatory conditions, including those active against another target associated with the disease, or a combination thereof. Such a combination may serve to increase efficacy, ameliorate other disease symptoms, decrease one or more side effects, or decrease the required dose of an inventive compound. The additional active ingredients may be administered in a separate pharmaceutical composition from a compound of the present invention or may be included with a compound of the present invention in a single pharmaceutical composition. The additional active ingredients may be administered simultaneously with, prior to, or after administration of a compound of the present invention.

[0108] Thus, a compound of the formula I-V may be used in combination with other antiproliferative compounds. Such antiproliferative compounds include, but are not limited to aromatase inhibitors; antiestrogens; topoisomerase I inhibitors; topoisomerase II inhibitors; microtubule active compounds; alkylating compounds; histone deacetylase inhibitors;

compounds which induce cell differentiation processes; cyclooxygenase inhibitors; MMP inhibittors; mTOR inhibitors; antineoplastic antimetabolites; platin compounds; compounds targeting/decreasing a protein or lipid kinase activity; anti- angiogenic compounds; compounds which target, decrease or inhibit the activity of a protein or lipid phosphatase; gonadorelin agonists; anti-androgens; methionine aminopeptidase inhibitors; bisphosphonates; biological response modifiers; antiproliferative antibodies; heparanase inhibitors; inhibitors of Ras oncogenic isoforms; telomerase inhibitors; proteasome inhibitors; compounds used in the treatment of hematologic malignancies; compounds which target, decrease or inhibit the activity of Fit- 3; Hsp90 inhibitors; kinesin spindle protein inhibitors; MEK inhibitors; leucovorin; EDG binders; antileukemia compounds; ribonucleotide reductase inhibitors; S-adenosylmethionine decarboxylase inhibitors; angiostatic steroids; corticosteroids; other chemotherapeutic compounds (as defined below); photosensitizing compounds.

[0109] Other chemotherapeutic compounds" include, but are not limited to, plant alkaloids, hormonal compounds and antagonists; biological response modifiers, preferably lymphokines or interferons; antisense oligonucleotides or oligonucleotide derivatives; shRNA or siRNA; or miscellaneous compounds or compounds with other or unknown mechanism of action.

[0110] Further, they may be used in combination with other cancer treatment approaches, including chemotherapy, radiotherapy, immunotherapy, surgical intervention, implants, or a combination of these. Chemical Synthesis

[0111] Exemplary chemical entities useful in methods of the invention will now be described by reference to illustrative synthetic schemes for their general preparation below and the specific examples that follow. Artisans will recognize that, to obtain the various compounds herein, starting materials may be suitably selected so that the ultimately desired substituents will be carried through the reaction scheme with or without protection as appropriate to yield the desired product. Alternatively, it may be necessary or desirable to employ, in the place of the ultimately desired substituent, a suitable group that may be carried through the reaction scheme and replaced as appropriate with the desired substituent. Each of the reactions depicted in Schemes is preferably run at a temperature from about room temperature (rt) to the reflux temperature of the organic solvent used. Unless otherwise specified, the variables are as defined above in reference to Formulas I-V.

Exemplary Synthesis:

[0112] The compounds described above can be synthesized as illustrated in Schemes 1. Some representative compounds are illustrated in Scheme lb, which depicts some suitable Het

Cyclo A: substituted benzenes, pyrindines, heterocyclic compounds

Cyclo A : substituted benzenes, pyrindines, heterocyclic amines

Cyclo B: substituted benzenes, pyrindine

Cyclo C: heterocyclic compounds

³ Reaction conditions: (a) substituted-o-aminonitrile, isothiocyanate 1.05 eq, reflux in the pyridine for 0.5-2 hours; (b) thiourea intermediates, benzylbromides 1.1 eq,

K₂C0₃ 2eq, stir in acetone at 50~60 deg for 2~3 hours.

Scheme 1: Generalized synthetic scheme

Z = S, O; n = 1 -5; A = bond, OO; CONH; or NHCO; etc.

R¹, R², R³, R = H, Me, Et, i-Pr, Propyl, Butyl, F, Br, CI, 1, OMe, NMe₂, NEt₂, N0₂, SMe, SEt, CF₃, CH₂COOEt, cyclopropyl, NHS0₂R, alkenyl, alkynyl, saturated/ unsaturated carbocyclic or heterocyclic ring (fused or not fused), etc. R and R' are as set forth for Formula I.

Scheme lb: Representative substituted and fused pyrazolo pyrimidine embodiments.

Detail procedures of target compounds and key intermediates

[0113] All cyclo-C compounds and isothiocyanates used were obtained from commercial sources (Sigma-Aldrich, Alfa Aesar). Most bromides were synthesized by our group member. Every final compound was characterized by LC-MS(API 150), and some active structures were analyzed by ^-NMR, ¹³C-NMR.

General Procedure for the Synthesis of Thiourea Intermediates:

[0114] A mixture of substituted-o-aminonitrile (lOmmol) and isothiocycanates (10.5mmol) were refluxed in pyridine (15mL) for over than half an hour, then light- yellow solids were appeared, meanwhile solvent decreased in a short time. The mixture were stirred for another 10 minutes, cooled to roomtemperature, filtered with Buchner funnel, wshed with 50ml EtOH, and dried in vacuum drying oven. The yield of pure products was high to 95%.

General Synthetic scheme:

[0115] Mixture of thiourea intermediates, bromides and K₂CO₃ (mol ratio, 0.5 mmol : 0.52 mmol : 1 mmol) in 3mL acetone, stirred and heated in 65 deg water bath for 2 hours. The reaction was completed indicated by TLC. Cool, add water 20mL and EA (ethyl acetate) 20 mL, extract, separate, collect and concentrate, the solid will crystallize, the last dry it after filtered. Yield was high to 85%.

General Procedure for the Synthesis of Intermediate Benzylbromo bromides [0116] Typical romides

[0117] Step 1. Methyl 5-methylnicotinate (10 mmol, 1.52g) was dissolved in 50 Ml dried THF, then cooled to 0 deg with ice-bath. LiAlH4 (16 mmol, 0.61g) was added in small portions during about 20 minutes. The mixture stirred for another 20 minutes until the reaction was completed, which monitored by TLC. 1.5 mL water was dropwised into the mixture to quench the reaction, then filtered, solution was concentrated to get the (5-methylpyridin-3-yl)methanol, Yield was high to 85%

[0118] Step 2. Alcohol (10 mmol) were refluxed in 10ml HBr (40% aq) for more than 5 hours, and monitored through TLC or LC-MS. After completed, mixture were heated in order to evaporator solvents (water and excess HBr) until the mixture became sticky. Cool the mixture, add acetone to the mixture, precipitated solid and filtered followed by drying. Yield was high to

80%.

[0119] Similar intermediate pyridine compounds can be synthesized using the above protocol. Some examples are provided in the Table below.

Synthesis of sulfone derivatives through oxidation (A429, A430, A434 and A484):

A372 A429

[0120] Mixture of A372, Na₂W0₄, H₂0₂ (mol ratio 1:2:2) were stirred in CH₃COOH at RT for3 hours, then add water and EA (ethyl acetate), extract, separate, collect, column chromatography and concentrate, the solid will crystallize, the last dry it after filtered. A429 was given in proper yields. This method were also applied in the preparation of A430, A434 and A484 from oxidation of A419, A348 and A424. Synthesis of sulfoxide derivatives through oxidation, A431, A433 and A485

A419 A431

[0121] Mixture of A419, NaB0₃.4H₂0 (mol ratio 1:1.1) were stirred in CH₃COOH at 40deg for3 hours, then add water and EA (ethyl acetate), extract, separate, collect and concentrate, the solid will crystallize, the last dry it after filtered. A431 was given in proper yields. This method was also applied in the preparation of A433 and A485 from oxidation of A348 and A424.

Synthesis of 2-bromo-l-(pyrrolidin-l-yl)ethanone

[0122] To 2-bromoacetyl bromide (10 mmol) in dry DCM (10 mL), pyrrolidine (10 mmol) in DCM (5 mL) was added drop- wise at -10 - -5°C (ice bath) with stirring. The addition was completed in ~ 50 min. TEA was then slowly added into the reaction mixture for 10 min. At this point, the ice bath was removed. The reaction mixture was warmed up to room temperature (rt) and continued stirring for 1 h. After removal of solid, the filtrate was collected and

concentrated. Acetone (20 mL) was added to this concentrated residue, giving a DCM/ Acetone solution of 2-bromo-l-(pyrrolidin-l-yl) ethanone (-0.35 mmol/ mL), which was stored at -20 °C for future use.

[0123] Similar intermediates can be prepared using the above protocol. Some examples are provided in the Table below.

Scheme lb Synthesis of substituted and fused pyrazolo pyrimidine derivatives

[0124] Substituted 5-amino pyrazole-4-carbonitrile 1 reacted with substituted

isothiocyanatobenzene 2 in pyridine stirred at refluxing temperature for 2 h providing the corresponding 4-imino-5 -substituted phenyl-4,5-dihydro-pyrazolo-[J,4-ii|pyrimidine-6(7H)- thione 3 in good yield. Thione 3 was alkylated with alkylation reagent halide R-X in acetone and K2CO₃ refluxing for 3 h giving the final desired product IV. Below is the synthesis of some representatives.

[0125] Compounds prepared according to the schemes described above may be obtained as single enantiomers, diastereomers, or regioisomers, by enantio-, diastero-, or regiospecific synthesis, or by resolution. Compounds prepared according to the schemes above may alternately be obtained as racemic (1 : 1) or non-racemic (not 1 : 1) mixtures or as mixtures of diastereomers or regioisomers. Where racemic and non-racemic mixtures of enantiomers are obtained, single enantiomers may be isolated using conventional separation methods known to one skilled in the art, such as chiral chromatography, recrystallization, diastereomeric salt formation, derivatization into diastereomeric adducts, biotransformation, or enzymatic transformation. Where regioisomeric or diastereomeric mixtures are obtained, single isomers may be separated using conventional methods such as chromatography or crystallization.

[0126] The following specific examples are provided to further illustrate the invention and various preferred embodiments.

Example 1 : Synthesis of 2-(4-imino-3-methyl-5-phenyl-4,5-dihydro- lH-pyrazolor3,4-d1

pyrimidin-6-ylthio-)- l-(pyrrolidin-l-yl)ethanone (A311 (9)):

[0127] The synthesis of 2-(4-imino-3-methyl-5-phenyl-4,5-dihydro-lH-pyrazolo[3,4-d] pyrimidin-6-ylthio-)-l-(pyrrolidin- l-yl)ethanone 8 was shown in Scheme 2.

Scheme 2 Synthesis of 2-(4-imino-3-methyl-5-phenyl-4,5-dihydro-lH-pyrazolo[3,4-d] pyr

9

[0128] Synthesis of 4-imino-3-methyl-5-phenyl-4,5-dihydro-lH-pyrazolo-[J,4- ii]pyrimidine-6(7H)-thione (7): Isothiocyanatobenzene 6 (1.16g, 8.6 mmol) was dissolved in 5 ml of dry pyridine. To this solution 3-methyl-5-amino pyrazole-4-carbonitrile 5 (1.0 g, 8.2 mmol) was added. The reaction mixture was heated up, stirred at refluxing for 2 h. A lot of yellow solid was precipitated. The reaction mixture was cooled to room temperature (rt). The yellow precipitation was collected, washed with warm ethanol and dried to yield the desired product 7 (1.45 g, 69%).

[0129] Synthesis of 2-(4-imino-3-methyl-5-phenyl-4,5-dihydro-lH-pyrazolor3,4-dl pyrimidin-6-ylthio-)-l-(pyrrolidin-l-yl)ethanone (A311 (9)): A mixture of 7 (200 mg, 0.78 mmol) and 2-bromo-l-(pyrrolidin-l-yl)ethanone (0.82 mmol, synthesized from 2-bromoacetyl bromide and pyrrolidine) in acetone (6 mL) with K₂CO₃ (216 mg, 1.56 mmol) was heated to refluxing and stirred for 3 h. The reaction mixture was cooled to room temperature and then water (10 mL) was added. White precipitation was collected, washed 3 times with water (to remove all inorganic salts), re-crystallized from MeOH to yield the desired product A311 (9) (M+l)⁺ = 369.5 (182 mg, white crystals, 63%). Example 2: Synthesis of 5-(2,6-difluorophenyl)-3-methyl-6-(pyridine-3-ylmethylthio)-lH- pyrimidin-4(5H)-imine (A348 (13)):

[0130] The synthesis of 5-(2,6-difluorophenyl)-3-methyl-6-(pyridine-3-ylmethylthio)-lH- pyrimidin-4

Acetone, refluxing, 3h A348 (13)

Scheme 3 Synthesis of 5-(2,6-difluorophenyl)-3-methyl-6-(pyridine-3-ylmethylthio)-lH- pyrimidin-4(5H)-imine A348 (13)

[0131] Synthesis of 5-(2,6-difluorophenyl)-4-imino-3-methyl-4,5-dihydro-lH-pyrazolo- [3,4-if]pyrimidine-6(7H)-thione (11): l,3-difluoro-2-isothiocyanatobenzene 10 (4.4 mmol) was dissolved in 3 ml of dry pyridine. To this solution 3-methyl-5-amino pyrazole-4-carbonitrile 5 (4.0 mmol) was added. The reaction mixture was heated up, stirred at refluxing for 2 h. A lot of yellow solid was precipitated. The reaction mixture was cooled to room temperature. The yellow precipitation was collected, washed with warm ethanol and dried to yield the desired product 11 (880 mg, 75%).

[0132] Synthesis of 5-(2,6-difluorophenyl)-3-methyl-6-(pyridine-3-ylmethylthio)- 1H- pyrimidin-4(5H)-imine (A348 (13)): A mixture of 11 (147 mg, 0.5 mmol) and commercially available 3-(bromomethyl)pyridine 12 (89 mg, 0.52 mmol) in acetone (3 mL) with K₂CO₃ (208 mg, 1.5 mmol) was heated to refluxing and stirred for 3 h. The reaction mixture was cooled to rt and then water (20 mL) was added. The mixture was extracted with ethyl acetate (3x 20 mL). The combined organic layers were washed with saturated NaHCC>3 solution and water, dried over Na₂S0₄ and concentrated down under reduced pressure to give crude product, which was further crystallized from MeOH to yield the desired product A348 (13) (M+l)⁺ = 385.5 (135 mg, white crystals, 70%). Example 3: Synthesis of 5-(2,6-difluorophenyl)-3-methyl-6-(pyridine-3-ylmethylsulfinyl) -1H- pyrimidin-4(5H)-imine (A433 (14)):

[0133] The Synthesis of 5-(2,6-difluorophenyl)-3-methyl-6-(pyridine-3-ylmethylsulfinyl) - lH-pyrimidin-4(5H)-imine (A433 (14)) was shown in Scheme 4.

Scheme 4 Synthesis of 5-(2,6-difluorophenyl)-3-methyl-6-(pyridine-3-ylmethylsulfinyl) - lH-pyrimidin-4(5H)-imine A433 (14)

[0134] Synthesis of 5-(2,6-difluorophenyl)-3-methyl-6-(pyridine-3-ylmethylsulfinyl) -1H- pyrimidin-4(5H)-imine (A433 (14)): A mixture of A348 (13) (54 mg, 0.14 mmol) and

NaB0₃4H₂0 (0.28 mmol) in CH₃COOH (3 mL) was stirred at 40°C for 3 h. The reaction mixture was cooled to rt, water (20 mL) was then added, and adjusted pH with 15% NaOH (aq) to pH >10. The mixture was extracted with ethyl acetate (3x 20 mL). The combined organic layers were washed with saturated NaCl aqueous solution and water, dried over Na₂S0₄ and concentrated down under reduced pressure to give crude product, which was crystallized from EtOAc/ Ether to yield the desired product A433 (14) (M+l)⁺ = 401.5 (20 mg, 36%).

Example 4: Synthesis of 5-(2,6-difluorophenyl)-3-methyl-6-(pyridine-3-ylmethylsulfonyl) -1H- pyrimidin-4(5H)-imine (A434(15)):

[0135] The Synthesis of 5-(2,6-difluorophenyl)-3-methyl-6-(pyridine-3-ylmethylsulfonyl ) - lH-pyrimidin-4(5H)-imine (A434(15)) was shown in Scheme 5.

Scheme 5 Synthesis of 5-(2,6-difluorophenyl)-3-methyl-6-(pyridine-3-ylmethylsulfonyl) - lH-pyrimidin-4(5H)-imine A434(15)

[0136] Synthesis of 5-(2,6-difluorophenyl)-3-methyl-6-(pyridine-3-ylmethylsulfony -1H- pyrimidin-4(5H)-imine (A434(15)): A mixture of A348 (13) (54 mg, 0.14 mmol), Na₂W0₄ (0.02 mmol) and ¾(¾ (0.2 mL ) in CH₃OH (3 mL) was stirred at room temperature for 3 h. Water (20 mL) was added. The mixture was extracted with ethyl acetate (3x 20 mL). The combined organic layers were washed with saturated NaHCC>3 solution and water, dried over Na₂S0₄ and concentrated down under reduced pressure to give crude product, which was crystallized from EtOAc/ Ether to yield the desired product A434(15) (M+l)⁺ = 417.5 (40 mg,

69 ).

Example 5: Synthesis of 5-(2,3,6-trifluorophenyl)-3-methyl-6-(5-methylpyridine-3- ylmethylthio)-lH-pyrimidin-4(5H)-imine (A424 (19)):

[0137] The synthesis of 5-(2,3,6-trifluorophenyl)-3-methyl-6-(5-methylpyridine-3- ylmethylthio)-lH-pyrimidin-4(5H)-imine (A424 (19)) was shown in scheme 6.

Scheme 6 Synthesis of 5-(2,3,6-difluorophenyl)-3-methyl-6-(5-methylpyridine-3- ylmethylthio)-lH-pyrimidin-4(5H)-imine A424 (19)

[0138] Synthesis of 5-(2,3,6-difluorophenyl)-4-imino-3-methyl-4,5-dihydro-lH-pyrazolo- r3,4-ti|pyrimidine-6(7H)-thione (17): l,2,4-trifluoro-3-isothiocyanatobenzene 16 (4.4 mmol) was dissolved in 3 ml of dry pyridine. To this solution 3 -methyl- 5 -amino pyrazole-4- carbonitrile 5 (4.0 mmol) was added. The reaction mixture was heated up, stirred at refluxing for 2 h. A lot of yellow solid was precipitated. The reaction mixture was cooled to room temperature. The yellow precipitation was collected, washed with warm ethanol and dried to yield the desired product 17 (890 mg, 71%).

[0139] Synthesis of 5-(2,3,6-trifluorophenyl)-3-methyl-6-(5-methylpyridine-3- ylmethylthio)-lH-pyrimidin-4(5H)-imine (A424 (19)): A mixture of 17 (156 mg, 0.5 mmol) and 3-(bromomethyl)-5-methyl pyridine 18 (97 mg, 0.52 mmol) (which was synthesized from commercially available methyl 5-methylnicotinate by reduction and bromination in a traditional way) in acetone (3 mL) with K₂CO₃ (208 mg, 1.5 mmol) was heated to refluxing and stirred for 3 h. The reaction mixture was cooled to rt and then water (20 mL) was added. The mixture was extracted with ethyl acetate (3x 20 mL). The combined organic layers were washed with saturated NaHC(¾ solution and water, dried over Na₂S0₄ and concentrated down under reduced pressure to give crude product, which was further crystallized from MeOH to yield the desired product A424 (19) (M+l)⁺ = 417.5 (145 mg, white crystals, 70%).

[0140] Using similar chemistry, the following compounds (A418 (20), A447 (21), and A448 (22)) were also synthesized:

Example 6: Synthesis of N-(5-((5-(2,6-difluorophenyl)-4-imino-3-methyl-4,5-dihydro-lH- pyrazolor3,4-dlpyrimidin-6-ylthio)methyl)pyridin-3-yl)formamide (A512 (23)):

A447 (21) A512 (23) Scheme 7 Synthesis of N-(5-((5-(2,6-difluorophenyl)-4-imino-3-methyl-4,5-dihydro-lH- pyrazolo[3,4-d]pyrimidin-6-ylthio)methyl)pyridin-3-yl)formamide (A512 (23))

[0141] Synthesis of N-(5 -((5 -( 2, 6-difluorophenyl)-4-imino-3 -methyl-4,5 -dihydro- 1 H- pyrazolor3,4-dlpyrimidin-6-ylthio)methyl)pyridin-3-yl)formamide (A512 (23)): A mixture of 21 (40 mg, 0.1 mmol), diiodine (5 mg, 0.04 mmol) and formic acid (5 mL) stirred on 70°C oil bath for 3 hours. The reaction mixture was cooled to rt followed by adding Na₂S₂C>3 (aq) 5mL and Na₂C(¾(aq) 20 mL, then extracted with ethyl acetate (25 mL x 3). The combined organic layers were collected, dried over Na₂S0₄ and concentrated down under reduced pressure to give crude product, which was further crystallized from MeOH to yield the desired product A512 (23) (M+l)+ = 428.1 (35 mg, white crystals, 82%).

Example 7: Synthesis of 5,6-bis(4-methoxyphenyl)-3-((pyridin-3-yl)methylthio)-l,2,4- triazine (A535 (25)):

24 A535 (25)

Scheme 8 Synthesis of 5,6-bis(4-methoxyphenyl)-3-((pyridin-3-yl)methylthio)-l,2,4- triazine (A535 (25)):

[0142] Synthesis of 5,6-bis(4-methoxyphenyl)-3-((pyridin-3-yl)methylthio)-l,2,4-triazine (A535 (25)): A mixture of 24 (98 mg, 0.3 mmol) and 3-(bromomethyl) pyridine 12 (55 mg, 0.33 mmol) in acetone (3 mL) with K₂CO₃ (138 mg, 1 mmol) was heated to refluxing and stirred for 3 h. The reaction mixture was cooled to rt and then water (20 mL) was added. The mixture was extracted with ethyl acetate (3x 20 mL). The combined organic layers were washed with saturated NaHC(¾ solution and water, dried over Na₂S0₄ and concentrated down under reduced pressure to give crude product, which was further crystallized from MeOH to yield the desired product A535 (25) (M+l)⁺ = 417 (145 mg, white crystals, 70%). Example 8: Synthesis of 5-(2, 6-difluorophenyl)-6-(pyridin-3-ylmethylthio)-lH-pyrazolor3,4- dlpyrimidin-4(5H)-one (A565 (27)):

A565 (27)

Scheme 9: 5-(2,6-difluorophenyl)-6-(pyridin-3-ylmethylthio)-lH-pyrazolo[3,4- d]pyrimidin-4(5H)-one (A565 (27))

[0143] Synthesis of 5-(2,6-difluorophenyl)-6-thioxo-6,7-dihydro-lH-pyrazolor3,4- dlpyrimidin-4(5H)-one(26). A mixture of ethyl 5-amino-lH-pyrazole-4-carboxylate (755 mg, 5 mmol) and l,3-difluoro-2-isothiocyanatobenzene (855 mg, 5 mmol) in dry toluene 5mL was refluxed for 6h, then cooled. Intermediate was precipitated spontaneously and were collected as pure products. A suspension of intermediate in 4% aqueous sodium hydroxide (4 mL) was refluxed until no more of the starting material could be detected by TLC or LC-MS (l-2h). After cooling, the solution was neutralized with 4 hydrochloricacid and then extracted with ethyl acetate (3 x20mL). After drying over anhydrous magnesium sulfate, the solvent was removed and the white solid residue was purified by flash column chromatography to give pure product (M+l)⁺ = 281 (1.226g, white powder, 87.6%).

[0144] The procedure for the Synthesis of 5-(2, 6-difluorophenyl)-6-(pyridin-3- ylmethylthio)-lH-pyrazolo[3,4-d]pyrimidin-4(5H)-one (A565 (27)) [(M+l)⁺ = 372.5] is the same as that of A512 (13).

[0145] Using similar chemistry, the following compound A566 (28), A567 (29), A568 (30), A569 (31), A570 (32) was also synthesized.

A569 (31) A570 (32)

Example 9: Synthesis of 4-imino-l-phenyl-5-(2,3,6-trifluorophenyl)-4,5-dihydro-lH- pyrazolor3,4-dlpyrimidine-6(7H)-thione (A575 (33)):

Synthesis of 4-imino-l-phenyl-5-(2,3,6-trifluorophenyl)-4,5-dihydro-lH-pyrazolor3,4- dlpyrimidine-6(7H)-thione (A575 (33)). A mixture of 5-amino-l -phenyl- lH-pyrazole-4 carbonitrile (218 mg, 1 mmol) and l,3,4-trifluoro-2-isothiocyanatobenzene (189 mg, 1 mmol) was refluxed for 2h,and then cooled. Intermediate was precipitated spontaneously and were collected as pure products (M+l)⁺ = 408 (325 mg, white solid, 80%). Then using the same procedure as that of A348 (13) to synthesis compound (A575 (33)) (M+l)⁺ = 498.

n erme a e

A575 (33) Example 10: Synthesis of 3-methyl-6-(p-tolylethvnyl)-5-(2,3,6-trifluorophenyl)-lH- pyrazolor3,4-dlpyrimidin-4(5H)-imine (A571 (36)):

Scheme 10, Synthesis of 3-methyl-6-(p-tolylethynyl)-5-(2,3,6-trifluorophenyl)-lH- pyrazolo[3,4-d]pyrimidin-4(5H)-imine (A571 (36))

[0146] Synthesis of 3-methyl-6-(p-tolylethynyl)-5-(2,3,6-trifluorophenyl)-lH-pyrazolor3,4- dlpyrimidin-4(5H)-imine (A571 (36)): A mixture of Ar-Br (107mg, 0.3 mmol), l-ethynyl-4- methylbenzene (38 mg, 0.33 mmol), Pd(PPh₃)₄ (35 mg, 0.03 mmol), Cul (6 mg, 0.03 mmol) and DIPA (8mg, 0.6 mmol) were refluxed in dry DMF 2mL for about 5 hours. The reaction was detected by TLC/LC-MS. After completed, extract with EA, remove solvent and purified through column chromatography on silica to give pure product (A571 (36)) (M+l)⁺ = 394 (65 mg, 55%).

Example 11: Synthesis of 5-amino-3-bromo-lH-pyrazole-4-carbonitrile (42) and 5-amino-3- chloro-lH-pyrazole-4-carbonitrile (43) and analogs A537 (44), A542 (45), A547 (46), A558 (47):

[0147] Synthesis of 5-amino-3-bromo-lH-pyrazole-4-carbonitrile (42) and 5-amino-3- chloro- 1 H-pyrazole-4-carbonitrile (43) : To a solution of 5-amino-lH-pyrazole-4-carbonitrile (540mg, 5 mmol) in dry MeCN 20mL, NBS (1209mg, 6.5 mmol) or NCS ( 920mg, 6.5 mmol) was added bathwise at room-temperature. The mixture was stirred forl2 hours, and detected by TLC/LC-MS. After completed, extract with EA, remove solvent and purified through column chromatography (EA:Hex, 1;1) on silica to give pure product(42) (M+l)⁺ = 187 (460mg, yellow solid, 49% yield), (43) (M+l)⁺ = 143 (320mg, white solid, 45% yield).

[0148] Using similar chemistry as Scheme 11 shows, the following compound A537 (44), A542 (45), A547 (46), A558 (47) were also synthesized.

Scheme 11: Synthesis of 5-((3-bromo-4-imino-5-(2,3,6-trifluorophenyl)-4,5-dihydro-lH- pyrazolo[3,4-d]pyrimidin-6-ylthio)methyl)pyridin-3-amine and derivates A537 (44), A542 (45), A547 (46), A558 (47)

Example 12: Synthesis of 5-(2,6-difluorophenyl)-6-((5-methylpyridin-3-yl)methylsulfonyl)-lH- pyrazolor3.4-dlpyrimidin-4(5H)-imine (A514 (48)) and (A513 (49)):

Scheme 12: Synthesis of 5-(2,6-difluorophenyl)-6-((5-methylpyridin-3-yl)methylsulfonyl)- lH-pyrazolo[3,4-d]pyrimidin-4(5H)-imine (A514 (48)) and 5-(2,6-difluorophenyl)- 6-((5-methylpyridin-3-yl)methylsulfinyl)-lH^yrazolo[3,4-d]pyrimidin-4(5H)-imine (A513 (49))

[0149] Synthesis of 5-(2,6-difluorophenyl)-6-((5-methylpyridin-3-yl)methylsulfonyl)-lH- pyrazolor3,4-dlpyrimidin-4(5H)-imine (A514 (48)) and (A513 (49)): Using similar chemistry as Schemes 4 and 5, compounds A514 (48), A513 (49) were also synthesized.

Biological screening and anticancer activity:

[0150] Examples demonstrating the anti-proliferation and anti-migration effects of the compounds of the invention are described as below.

In vitro cell-based screening using real-time cell electronic sensing (RT-CES) system

[0151] The heterocyclic compounds in the present invention are developed for the anticancer activities for cancer cells with certain molecular targets, i.e. , cMet. The anticancer efficacy of these heterocyclic compounds and their analogues described above were

preliminarily screened in vitro using a penal of cMet- addicted cancer cell lines by real time electronic cell sensing (RT-CES) system from ACEA Biosciences, Inc. (or xCELLigence system from Roche Applied Sciences/ ACEA Biosciences Inc.) , which provides dynamic cell response information after exposing to cMet inhibitors.

[0152] The details of this cell electronic sensing technology, called real-time cell electronic sensing (RT-CES^®) and associated devices, systems and methods of use are described in United States patent number 7,732,127; patent number 7,192,752; patent number 7,459,303; patent number 7,468,255; patent number 7,470,533; patent number 7,560,269; United States provisional application number 60/397,749, filed on July 20, 2002; United States provisional application number 60/435,400, filed on December 20, 2002; United States Provisional application 60/469,572, filed on May 9, 2003, PCT application number PCT/US03/22557, filed on July 18, 2003; PCT application number PCT/US03/22537, filed on July 18, 2003; PCT application number PCT/US04/37696, filed on November 12, 2004; PCT application number PCT/US05/04481, filed on February 9, 2005; United States patent application number

10/705,447, filed on November 10, 2003; United States patent application number 10/705,615, filed on November 10, 2003; United States patent application number 10/987,732, filed on November 12, 2004; United States patent application number 11/055,639, filed on

February 9, 2005, each of which is incorporated by reference. Additional details of RT-CES technology is further disclosed in United States provisional application number 60/519,567, filed on November 12, 2003, and United States provisional application number 60/542,927, filed on February 9, 2004, United States provisional application number 60/548,713, filed on February 27, 2004, United States provisional application number 60/598,608, filed on

August 4, 2004; United States provisional application number 60/598,609, filed on

August 4, 2004; United States provisional application number 60/613,749, filed on

September 27, 2004; United States provisional application number 60/613,872, filed on September 27, 2004; United States provisional application number 60/614,601, filed on September 29, 2004; United States provisional application number 60/630,071, filed on November 22, 2004; United States provisional application number 60/630,131, filed on

November 22, 2004, each of which is incorporated herein by reference.

[0153] For measurement of cell-substrate or cell-electrode impedance using RT-CES technology, microelectrodes having appropriate geometries are fabricated onto the bottom surfaces of microtiter plate or similar device, facing into the wells. Cells are introduced into the wells of the devices, and make contact to and attach to the electrode surfaces. The presence, absence or change of properties of cells affects the electronic and ionic passage on the electrode sensor surfaces. Measuring the impedance between or among electrodes provides important information about biological status of cells present on the sensors. When there are changes to the biological status of the cells analogue, electronic readout signals are measured automatically and in real time, and are converted to digital signals for processing and analysis.

[0154] In a RT-CES system, a cell index is automatically derived and provided based on measured electrode impedance values. The cell index obtained for a given well reflects: 1) how many cells are attached to the electrode surfaces in this well; 2) how well cells are attached to the electrode surfaces in this well. Thus, the more the cells of same type in similar

physiological conditions attach the electrode surfaces, the larger the cell index. And, the better the cells attach to the electrode surfaces (e.g., the cells spread-out more to have larger contact areas, or the cells attach tighter to electrode surfaces), the larger the cell index. Impedance- based TCRP (time-dependent cell response profiling) has shown to be extremely useful in predicting the mechanism of compound action (Abassi, Y. A., et ah, Chem. Biol. (2009) 16:712- 723). Oncogene addiction refers to the acquired dependency of cancer cells on a single cellular pathway for survival or sustained proliferation, despite the fact that such cells have accumulated numerous genetic alterations. We have discovered that the c-Met-addicted cell lines would produce a signature impedance-based TCRP, when treated with positive-control c-Met inhibitors.

[0155] Through the use of the RT-CES system, the heterocyclic compounds described in the examples above have been shown to produce a similar impedance-based TCRP on RT-CES system to that generated by positive control inhibitors (Figure 1-2). The EC50 calculated based on the short-term TCRP correlates well with the IC50 calculated based other cell based assay (e.g. , Western and ELISA analysis evaluating cMet inhibition). These compounds have also been shown to inhibit HGF (hepatocyte growth factor) -induced cell migration (Figure 4-5). In addition, these compounds showed no or negligible effects on non-cMet addicted cancer cell lines (Figure 3).

Example 13 Bioactivity of heterocyclic compounds on cMet- addicted cell lines as determined using real-time cell electronic sensing (RT-CES) system

[0156] H1993 and GTL16 have previously been reported as cMET addicted cell lines (Benvenuti, S., et ah, Cancer Res. (2011) 71 : 1945-1955). We used these two cell lines to assay specific inhibitory effects of the heterocyclic compounds described in the examples above on cMet. H1993 and GTL16 were seeded into wells of 96 well E-plate devices (Roche) with initial seeding density of 10,000 cells per well and were pre-incubated in incubator under standard cell culture condition for about 24 hours. The heterocyclic compounds at different concentration in DMSO were added into wells following the incubation period. Negative control wells were treated with buffer solutions containing DMSO only. Figure 1 shows the TCRP (time- dependent cell response profiling) of c-Met addicted HI 993 cells to the treatment of various heterocyclic compounds. The dose-dependent rapid impedance (corresponding to normalized cell index in the y-axis) increase and TCRP are the same as those observed for H1993 cells when treated with cMet inhibitor control compounds (results not shown). The cell index was normalized against the cell index values at a time point just before compound addition. The calculated effective concentrations EC50 (2 hr post compound treatment) in this specific experiment is 220 nM, 179 nM, 217 nM and 154 nM for compound A424 (19),

compound A418 (20), compound A447 (21) and compound A448 (22), respectively. Figure 2 shows the TCRP (time-dependent cell response profiling) of c-Met addicted GTL16 cells to the treatment of various heterocyclic compounds. The dose-dependent rapid impedance

(corresponding to normalized cell index in the y-axis) increase and TCRP are the same as those observed for GTL16 cells when treated with cMet inhibitor control compounds (results not shown). The calculated effective concentrations EC50 (2 hr post compound treatment) in this specific experiment is 137 nM, 141 nM, 138 nM and 109 nM for compound A424 (19), compound A418 (20), compound A447 (21) and compound A448 (22), respectively. The EC50 calculated based on the short-term TCRP correlates well with the IC₅₀ calculated based other cell based assay (EXAMPLE 17).

[0157] Results for the compounds tested in these assays are presented in Table 1-11 as an average of results obtained. The EC5₀S (in H1993) toward cMet inhibition for selected compounds of this invention are listed in Tables 1-11. Compounds were tested in free base form. Where activity is shown as greater than (>) a particular value, the value is the highest concentration tested.

Compound No. Cyclo A Cyclo B Cyclo C EC50

A348 (13) H 1,5-di-F R=Me 0.5-1.3uM

A351 H 1,4-di-F R=Me >20uM

A352 H H R=Et >20uM

A353 H 3-OMe R=Me 12.0uM

A354 H 1,3,4-tri-F R=Me >20uM

A356 H 1-F R=Et >20uM

A357 1,4-di-F R=Me >20uM

A358 1,4-di-F R=Me >20uM

A359 1,3,4-tri-F R=Me >20uM

A361 2,3,4-tri-F R=Me >20uM

A362 2,3,4-tri-F R=Me >20uM

A363 2,4-di-F R=Me >20uM

A364 2,4-di-F R=Me >20uM

A365 2,4-di-F R=Me >20uM

A366 2,3-di-F R=Me >20uM

A367 2,3-di-F R=Me >20uM

Compound Cyclo A Cyclo B Cyclo C EC50

No.

A368 2-F,4-Me R=Me >20uM

A373 H 1-F R=Me >20uM

A374 H 2-F R=Me >20uM

A375 H 3-Me R=Me >20uM

A376 H 1,3-di-F R=Me >20uM

A377 H l,5-2Me R=Me >20uM

A379 H 1-Cl R=Me >20uM

A382 2-Cl 1,5-di-F R=Me 12.7uM

A383 H 1-F,4-Me R=Me 18.4uM

A385 H 1-F.4-C1 R=Me >20uM

A386 H 1-Me,3-F R=Me 11.7uM

A389 H N=\ "^L R=Me >20uM

A391 H 1-C1.3-F R=Me >20uM

A393 H 1-OMe R=Me >20uM

A395 H 1-N0₂ R=Me >20uM

A396 H 3-Cl R=Me >20uM

Compound Cyclo A Cyclo B Cyclo C EC50

No.

A409 H 1,2,5-tri-F R=Me 0.25-0.55uM

A411 H 1.5-2C1 R=Me 4.4uM

A413 H 1-F.4-C1 R=Me >20uM

A414 H l,4-20Me R=Me >20uM

A415 H 1-F,4-Br R=Me >20uM

A416 H 1-F.5-C1 R=Me 4.6uM

A417 ^¾ r^F 1-F.5-C1 R=Me >20uM

A418 (20) 3-Me 1,5-di-F R=Me 0.16-0.51uM

A419 3-Me H R=Me 5.9uM

A422 1,5-di-F R=Me 12.4uM

A423 1,5-di-F R=Me >20uM

A424 (19) 3-Me 1,2,5-tri-F R=Me 0.13-0.64uM TABLE 4

Compound Cyclo A Cyclo B Cyclo C EC50

No.

A425 1,2,5-tri-F R=Me 18.5uM

A437 3-F 1,5-di-F R=Me 3.4uM

A438 3-Br 1,5-di-F R=Me 2.8uM

A439 3-Cl 1,5-di-F R=Me 2.7uM

A440 3-CF₃ 1,5-di-F R=Me 11.7uM

\ /

A441 1,5-di-F R=Me >20uM

A442 3-OMe 1,5-di-F R=Me lO.luM

A443 3-OH 1,5-di-F R=Me 4.4uM

A444 3-cyclopropyl 1,5-di-F R=Me >20uM

A445 3-Et 1,2,5-tri-F R=Me 2.9uM

A446 3-Et 1,5-di-F R=Me 3.5uM

Compound Cyclo A Cyclo B Cyclo C EC50 No.

2,3-CH==CH-

A452 1,5-di-F R= =Me >20uM

CH==CH

2,3-CH==CH- 1,2,5-tri-

A453 R= =Me >20uM

CH==CH F

A458 3-NH₂,4-Me 1,5-di-F R= =Me >20uM A459 2- NH₂ 1,5-di-F R= =Me >20uM

A460 1,5-di-F R= =Me >20uM

A461 2,3-2Me 1,5-di-F R= =Me >20uM A462 1-NH₂,3-Me 1,5-di-F R= =Me >20uM

1,2,5-tri-

A463 2,3-2Me R= =Me >20uM

F

A464 3- C0₂Me 1,5-di-F R= =Me >20uM

1,2,5-tri-

A465 1- NH₂,3-Me R= =Me 9.5uM

F

A467 2- Me 1,5-di-F R= =Me >20uM

1,2,5-tri-

A468 2- Me R= =Me >20uM

F

A469 4- Me 1,5-di-F R= =Me >20uM A470 3- NHMe 1,5-di-F R= =Me 3.6uM A471 1,5-di-F R= =Me >20uM

A472 1,5-di-F R= =Me 14.9uM

TABLE 6

Compound Cyclo A Cyclo Cyclo EC50

No. B C

A473 3-Ph 1,5-di-F R=Me >20uM A474 3-NHCOCH3 1,5-di-F R=Me 1.5uM

A475 1,5-di-F R=Me 2.6uM

A476 1,5-di-F R= =Me >20uM

A477 3-COOH 1,5-di-F R= =Me >20uM

A479 3-CN 1,5-di-F R= =Me >20uM

A480 3-NHCONH2 1,5-di-F R= =Me 2.4uM

A481 H 1,5-di-F R= =H 8.8uM

A482 3-Me 1,5-di-F R= =H 3.5uM

A483 3-CH2NH2 1,5-di-F R= =Me >20uM

TABLE 7

Compound Cyclo A Cyclo B Cyclo C

No.

A299 CH- H R=H >20uM

A300 H R=H 13.4uM

A301 t>i- 3-Cl R=H >20uM

A303 3-Me R=H >20uM

A304 Ol- 3-Me R=H >20uM

A306 Oh H R=H >20uM

A313 MeO-Q-f- H R=Me >20uM

>20uM

A314 H R=Me

TABLE 8

Compound Cyclo A Cyclo B Cyclo C EC50

No.

A315 3-Br R=Me >20uM

A316 H R=Me >20uM

A377 CM- 3-Br R=Me >20uM

A318 σι- 3-Br R=Me >20uM

A319 C^N- - 1-F R=Me 10.8uM

A320 C^N-I- 1-F R=Me >20uM

A321 C^N- - 2-F R=Me 15.0uM

A322 C^N-I- 3-F R=Me 15.1uM

A323 C^N-I- 2-F R=Me >20uM

A324 H R=Me >20uM

A325 ^ipr !- 3-F R=Me >20uM

A326 3-Cl R=Me 4.4uM

A327 ^ipr -h 3-Cl R=SMe >20uM

'Pr ^s

2- Cl,

A328 C^N-I- R=Me About 1

3- F

A329 2-Cl, 3-F R=Me >20uM

^■Pr ⁵

A330 C^N-I- 1,3-di-F R=Me >20uM

TABLE 9

Compound Cyclo A Cyclo B Cyclo C EC50 No.

A331 1,3-di-F R= :Me >20uM

^■Pr ⁶

Et

A332 1,3-di-F R= =Me >20uM

E.- -

A333 ^ipr -h 1,5-di-F R= =Me >20uM

Et

A334 1,5-di-F R= =Me >20uM

Ε,·Ν- A335 0-4- 1,5-di-F R= =Me 11.9uM A336 0.4- 1,4-di-F R= =Me 13.1uM

A338 Q^N- - H R= =Me >20uM A341 0-4- 3-Me R= =Me >20uM A342 0-4- 2-CF₃ R= =Me >20uM A343 0-4- 3-CF3 R= =Me >20uM A344 0-4- 3-OMe R= =Me 9.4uM

A350 0-4- H R= =Et >20uM

TABLE 10

Compound Cyclo A Cyclo B Cyclo C EC50 No.

A370 Oh 2,3-di-F R=Me 12.1uM

A378 Oh l,5-2Me R=Me >20uM

A380 1-2C1 R=Me 15.6uM

A381 1,5-di-F R=Me >20uM

A384 CN-I- 1-F.4-C1 R=Me >20uM

A387 CN-I- 3-Br R=Me >20uM

A390 Oh 1-C1.3-F R=Me >20uM

A392 Oh 1-OMe R=Me 15.1uM

A394 CN-I- 2-N0₂ R=Me >20uM

A397 Oi- 3-Cl >20uM

V-

/

A398 0-4- 3-Cl >20uM

A404 1,5-di-F >20uM

V-

A405 Oi- 1,5-di-F R=Me >20uM TABLE 11

Compound Cyclo A Cyclo B Cyclo C EC50

Example 14 Bioactivity of heterocyclic compounds on cMet-non- addicted cell lines as

determined using real-time cell electronic sensing (RT-CES) system

[0158] To examine whether the heterocyclic compounds produce any bioactivity effects on cMet-non-addicted cell lines and to verify that the effect of these compounds on cMet- addicted cells was specific, A549 cells were treated with various heterocyclic compounds described above. A549 were seeded into wells of 96 well E-plate devices (Roche) with initial seeding density of 5,000 cells per well and were pre-incubated in incubator under standard cell culture condition for about 24 hours. The heterocyclic compounds at different concentration in DMSO were added into wells following the incubation period. Negative control wells were treated with buffer solutions containing DMSO only. Figure 3 shows the TCRP of cMet-non- addicted A549 cells to the treatment of various heterocyclic compounds. No obvious effects were observed for A549 cells when treated with these compounds (compound A348 (13), compound A424 (19), compound A418 (20), compound A447 (21) and compound A448 (22)). This suggests that their effect on cMet addicted cells was specific to cMet inhibition.

Example 15 Inhibitory effect of heterocyclic compounds on HGF-mediated migration of

A431 cells as determined using real-time cell electronic sensing (RT-CES) system

[0159] To validate and confirm the cMet-inhibitory effects of various heterocyclic compounds described above, HGF-mediated migration of A431 cells was performed in the presence of different heterocyclic compounds. The technical details, instrument, devices of cell migration/invasion assays with real-time cell electronic sensing technology can be found in United States patent number 7,459,303. Briefly, A431 cells were starved for 6 hr and then incubated with compounds for 1 hr before seeding to fibronectin precoated CIM device (Roche). HGF at 50 ng/ml was used to mediate migration of A431 from upper chamber to lower chamber. The time course of the migration was monitored.

[0160] Table 12 below shows IC₅₀ values derived from the impedance response profile of HGF-mediated migration of A431 cells in the presence of different compounds described in the present invention. Figure 4 shows that these heterocyclic compounds inhibited HGF-mediated A431 migration in a dose-dependent manner. The positive control compound, PF02341066 (HGFi) inhibited A431 migration in a similar dose-dependent manner. These results support the hypothesis that these compounds are cMet (HGFR) inhibitors. The IC₅₀ values were derived from the dose responses at 10, 20 and 30 hrs after migration experiments started.

Table 12

Example 16 Inhibitory effect of heterocyclic compounds on HGF-mediated migration of

HUVECs as determined using real-time cell electronic sensing (RT-CES) system

[0161] To further validate and confirm the cMet-inhibitory effects of various heterocyclic compounds described above, HGF-mediated migration of HUVECs (human umbilical vein endothelial cells) was performed in the presence of a series of concentrations of different heterocyclic compounds. The technical details, instrument, devices of cell migration/invasion assays with real-time cell electronic sensing technology can be found in United States patent number 7,459,303. Briefly, HUVECs were starved for 6 hr and then incubated with compounds for 1 hr before seeding to fibronectin precoated CIM device (Roche). HGF at 50 ng/ml was used to mediate migration of HUVECs from upper chamber to lower chamber. The time course of the migration was monitored. [0162] Table 13 below lists IC50 values derived from the impedance response profile of HGF-mediated migration of HUVECs in the presence of different compounds described in the present invention. Figure 5 shows that these heterocyclic compounds inhibited HGF-mediated HUVECs migration in a dose-dependent manner. The positive control compound, PF02341066 (HGFi) inhibited HUVECs migration in a similar dose-dependent manner. These results further support the notion that these compounds are cMet (HGFR) inhibitors. The IC₅₀ values were derived from the dose responses at 6 hrs and 12 hrs after migration experiments started.

Table 13

Example 17 Heterocyclic compounds show inhibitory effect on cMet phosphorylation both in vivo and in vitro

[0163] To further validate and confirm the cMet-inhibitory effects of various heterocyclic compounds described above, we employed other approaches (not RT-CES based assays). We first tested if compound 13 inhibited cMet phosphorylation in A549 cells stimulated by HGF. Briefly, A549 cells were first treated with compounds in serum free medium for 2 hours, then stimulated with 100 ng/mL HGF for 30 min. Cells were lysed and the lysates were used for detection of the total phospho-cMet using an ELISA kit (R&D system, DYC2480-2). The results (Figure 6a) show that compound A348 (13) inhibited cMet phosphorylation in a dose dependent manner. The calculated IC₅₀ is 332.5 nM. In comparison, the calculated IC₅₀ for the positive control (HGFi, PF02341066) is 2.25 nM.

[0164] We also evaluated the inhibitory effect of the heterocyclic compounds on constitutively active cMet in GTL16 (one of cell lines used in RT-CES assay) using Western analysis. Figure 6b shows that cMet phosphorylation at Y1234/1235 was inhibited by compound 13 treatment in a dose-dependent fashion. The intensities of the phospho-cMet bands were quantified and used for IC₅₀ calculation. The calculated IC₅₀ for compound A348 (13) is 563 nM, similar to that derived from ELISA assay, as well as migration assay (Table 12, compound A348 (13)). In addition, we evaluated the potency of compound A447 (21), compound A418 (20) and compound A424 (19) in GTL16 using Western analyses. The calculated IC₅₀ is 38.8 nM, 74.7 nM and 36.2 nM for compound A 447 (21), compound A418 (20) and compound A424 (19) and, respectively. Furthermore, we tested the activities of the heterocyclic compounds on the enzymatic activities of purified cMet protein (experiments were performed at Reaction Biology Corporation). At 10 μΜ ATP concentration, the IC₅₀ is 494.9 nM, 10.8 nM, 4.01 nM and 1.21 nM for compound A348 (13), compound A447 (21), compound A418 (20) and compound A424 (19), respectively. The potency ranking agrees well with that derived from our RT-CES based assay. Taken together, there is strong evidence that the heterocyclic compounds described herein are cMet inhibitors.

EXAMPLE 18 Heterocyclic compounds show inhibitory effect on other kinase targets in vitro

[0165] KINOMEscan is the industry's most comprehensive high-throughput system for screening compounds against large number of human kinases (Fabian et al. (2005) Nat.

Biotechnol. 23, 329; Karaman et al. (2008) Nat. Biotechnol. 26, 127). To evaluate the activities of these heterocyclic compounds on other kinase targets, we have performed KINOMEscan on all 442 available kinases at Ambit Biosciences. Table 14 lists the 42 kinases that were inhibited by cpd20 (10 μΜ) by more than 65%.

Table 14

[0166] To confirm these results using activity based assay, we also tested the effects of compound A418 (20), compound A424 (19) and compound A447 (21) on the kinase activities of Aurora kinases, Jak2 and ROS l . At 10 uM concentration, all these kinase are effectively inhibited. Table 15 lists the % enzyme activities of these kinases in the presence of 10 μΜ compound A418 (20), compound A424 (19) and compound A447 (21) at a given ATP concentration.

Table 15

EXAMPLE 19 Heterocyclic compounds show inhibitory effect on RQS 1

phosphorylation in vivo

[0167] ROSl transcripts have been detected in several glioblastoma cell lines, e.g., U-118 MG, U-138 MG and SW-1088, but not in A-172 and A-382 (Fabian et al. (1987) Proc. Nati. Acad. Sci. USA 84, 9270-9274). We have tested the effect of these heterocyclic compounds on ROSl phosphorylation in these glioblastoma cell lines. The results from Western analyses confirm that ROSl was expressed in U138 MG, not in A-172. In addition, the Western results show that at 10 μΜ concentration, compound A348 (13), compound A418 (20), compound A424 (19) and compound A447 (21) inhibited ROSl phosphorylation after 1 hour compound treatment (Figure 7).

Claims

CLAIMS:

1.

Formula la Formula lb where in a ring indicates the ring is an aromatic or heteroaromatic ring;

W¹, W² W³ _, W⁴ are each independently absent, N, NH, NR¹, O, S, CH, or CR²,

provided at least one of W¹, W² W³ _, and W⁴ is CH or CR²,

not more than one of them is absent,

and not more than one of them is O or S;

R' is Ci-8 alkyl, C₂-8 alkenyl, C₂-8 alkynyl, aryl, or heteroaryl, wherein each alkyl, alkenyl, alkynyl, aryl and heteroaryl is optionally substituted;

Z is S, SO, S0₂, SO₂NH, SO₂NR³, NHSO₂, NR¹, CR¹]*², NR¹, or O;

Y is (CHR⁴)_n wherein n = 1-5 or a C3-C7 carbocyclic or aromatic divalent moiety;

A is a bond (absent), C(=0), NH, NR⁵, C(0)NH, C(0)NR⁶, C(S)NH, C(S)NR⁶, NHC(O), NR⁶C(0), NHSO₂, NR⁶S0₂, SO₂NH, SO₂NR⁶, or OP(=0)(OR⁷);

Het is substituted or unsubstituted Ci_-8 alkyl, C₂-8 alkenyl, C₂-8 alkynyl, aryl, fused aryl, heteroaryl, fused heterocycle, a saturated or unsaturated carbocyclic ring or a heterocyclic ring, or a group of the formula -(CH₂)_q-Q where q is 1-4 and Q is an aryl, heteroaryl, cycloalkyl or heterocycle and Q is optionally substituted,

where each Het is optionally substituted and may contain a heteroatom selected from N, O and S in place of a ring or chain carbon atom of Het;

each of R¹, R², R³, R⁴, R⁵, R⁶, and R⁷ is independently selected from H, OH, Halo, NHR, NRR, OR, SR, COOR, C(=0)R, CN, CF₃, OCF₃, N0₂, OC(0)R, S0₃R, PO3R2,

CR(COOR)₂;

each R is independently selected from H, halo, OR^x, SR^X, C0₂R^x, C(0)NR^x ₂, C(=0) R^x, CN, CF₃, OCF3, N0₂, NR^X2, OCOR^x, S0₃R^x, P0₃R^x ₂, and CH(COOR^x)₂;

where each R^x is H or C1-C4 alkyl or C1-C4 haloalkyl,

or R can be substituted or unsubstituted Ci_-8 alkyl, C₂-8 alkenyl, C₂-8 alkynyl, aryl, fused aryl, heteroaryl, fused heterocycle, a C3-8 carbocyclic ring or a C₃-8 heterocyclic ring, saturated or unsaturated, wherein each Ci_-8 alkyl, C3_g cyclic alkyl, C2-8 alkenyl, C2-8 alkynyl can optionally contain a heteroatom selected from N, O and S in place of a carbon atom; p is 0-3;

each R¹⁰ is H or acyl;

and two R, R , R , R , R , R , R , or R on the same or adjacent atoms can optionally be linked together to form a 3-8 membered ring that can contain up to two heteroatoms selected from N, O and S as ring members and which is optionally substituted;

or a pharmaceutically acceptable salt thereof.

2. The compound of claim 1, wherein W⁴ is CR² and W³ is a bond.

3. The compound of any one of the preceding claims, wherein W¹ is NR¹.

4. The compound of any of the preceding claims, wherein R¹⁰ is H.

5. The compound of any of the preceding claims, wherein Z is S, SO, SO2, SO2NH, SO2NR³, NHSO2, NR¹, CR^² NR¹, or O;

6. The compound of any one of the preceding claims, wherein Y is (0¾)ι^.

7. The compound of any of the preceding claims, wherein A is a bond or C(=0).

8. The compound of any one of the preceding claims, Het is a 5 or 6 membered aromatic or heteroaromatic ring.

9. The compound of any one of the preceding claims, wherein Het is phenyl or pyridyl and is optionally substituted with up to three R groups.

10. The compound of any one of the preceding claims, wherein W⁴ is CR², wherein R² is H, CI -4 alkyl, or CI -4 haloalkyl.

11. The compound of any of the preceding claims, wherein R' is optionally substituted phenyl.

12. The compound of any one of the preceding claims, wherein W² is N. A com ound of Formula Ila and Formula lib

Ila lib

wherein:

W¹ is NR¹, O or S;

W² is N or CR²;

Ar is a 5-6 membered aryl or heteroaryl ring that can be substituted or unsubstituted; and

R², R¹⁰, Z, Y, A, and Het are as defined in claim 1 ;

or a pharmaceutically acceptable salt thereof.

14. The compound of claim 13, wherein R¹⁰ is H.

15. The compound of claim 13 or 14, wherein W¹ is NR¹.

16. The compound of any one of the claims 13-15, wherein W¹ is O or S.

17. The compound of any one of the claims 13-16, wherein W² is N or CR².

18. The compound of any one of the claims 13-17, wherein Z is S, SO, SO2, SO2NH, SO2NR³, NHSO2, NR¹, CR^² NR¹, or O;

19. The compound of any one of the claims 13-18, wherein Y is (CH^i^.

20. The compound of any one of the claims 13-19, wherein A is a bond or C(=0).

21. The compound of any one of the claims 13-21, Het is a 5 or 6 membered aromatic or heteroaromatic ring.

22. The compound of any one of the claims 13-21, wherein Het is phenyl or pyridyl and is optionally substituted with up to three R groups.

23. The compound of any one of the claims 13-22, wherein R² is H, Ci_₄ alkyl, or Ci_ 4 haloalkyl.

24. The compound of any one of the claims 13-21, wherein Ar is optionally substituted phenyl.

25. The compound of claim 13,

whe

wherein the wavy line across one non-cyclic bond indicates the bond that connects each Het group to A; where in a ring indicates the ring is an aromatic or heteroaromatic ring;

each W and W is independently a bond, C, CH, CH₂, CHR¹², CR¹², N, NH, NR¹², S, O, C=0, or C=S,

where each R is independently selected from H, halo, OR^x, SR^X, C0₂R^x, C(0)NR^x ₂, C(=0) R^x, CN, CF₃, OCF₃, N0₂, NR^X2, OCOR^x, S0₃R^x, P0₃R^x ₂, and CH(COOR^x)₂, where each R^x is H or C1-C4 alkyl or C1-C4 haloalkyl,

or R can be substituted or unsubstituted Ci_g alkyl, C₂_g alkenyl, C₂_g alkynyl;

and each R¹² is H or C1-C4 alkyl or C1-C4 haloalkyl,

or a pharmaceutically acceptable salt thereof.

26. The compound of claim 25, wherein R is selected from H, halo, OR^x, S(0)_tR^x, C0₂R^x, C(0)NR^X2, C(=0) R^x, CN, CF₃, OCF₃, N0₂, NR^X ₂, OCOR^x, S0₃R^x,

P0₃R^x ₂, CH(COOR^x)₂,

where each R^x is H or C1-C4 alkyl or C1-C4 haloalkyl, and each t is 0-2.

27. The compound of claim 25, wherein R is Ci_g alkyl, C3_g cyclic alkyl, C2-8 alkenyl, C2-8 alkynyl, an aryl, heteroaryl, a carbocyclic ring or a heterocyclic ring, each of which may contain a heteroatom selected from N, O and S in place of a carbon atom of the alkyl, alkenyl, alkynyl group.

28. A com ound of Formula Ilia and Formula Illb:

Ilia Illb

wherein:

W¹ is NR¹, O or S;

Z is S, SO, S0₂, SO2NH, SO2NR³, NHSO2, NR¹, C^R² NR¹,

Y is -CH₂- or -CH2-CH2- or -CH(Me)-;

A is a bond or C(=0);

R¹⁰ is H;

Ph is an optionally substituted phenyl group;

R, R¹, R², R³, and Het are as defined in claim 1 ;

or a pharmaceutically acceptable salt thereof.

29. The compound of claim 28,

where

wherein the wavy line across one non-cyclic bond indicates the bond that connects each Het group to A; where in a ring indicates the ring is an aromatic or heteroaromatic ring; each W and W is independently a bond, C, CH, CH₂, CHR , CR , N, NH, NR , S, O, C=0, or C=S,

where each R is independently selected from H, halo, OR^x, SR^X, C0₂R^x, C(0)NR^x ₂, C(=0) R^x, CN, CF₃, OCF₃, N0₂, NR^X2, OCOR^x, S0₃R^x, P0₃R^x ₂, and CH(COOR^x)₂, where each R^x is H or C1-C4 alkyl or C1-C4 haloalkyl,

or R can be substituted or unsubstituted Ci_-8 alkyl, C2-8 alkenyl, C2-8 alkynyl;

and each R¹² is H or C1-C4 alkyl or C1-C4 haloalkyl,

or a pharmaceutically acceptable salt thereof.

30. The compound of claim 29, wherein Het is phenyl or pyridyl and is optionally substituted with up to three groups selected from C1-C4 alkyl, C1-C4 haloalkyl, halo, OR^x, SR^X, C0₂R^x, C(0)NR^X2, C(=0) R^x, CN, CF₃, OCF₃, N0₂, NR^X ₂, OCOR^x, S0₃R^x, P0₃R^x ₂, and CH(COOR^x)₂, where each R^x is H or C1-C4 alkyl or C1-C4 haloalkyl.

A c

wherein:

Het is as defined in claim 1 ;

n is 1-4;

R¹, R² and R³ is independently selected from H, OH, Halo, NHR', NR'R', OR', SR', COOR', C(=0)R', CN, CF₃, OCF₃, N0₂, OC(0)R', S0₃R', P0₃R'₂,

CR'(COOR')₂; each R' is independently selected from H, halo, OR^x, SR^X, C0₂R^x, C(0)NR^X2, C(=0) R^x, CN, CF₃, OCF₃, N0₂, NR^X ₂, OCOR^x, S0₃R^x, P0₃R^x ₂, and

CH(COOR^x)₂;

where each R^x is H or C1-C4 alkyl or C1-C4 haloalkyl;

or a pharmaceutically acceptable salt thereof.

32. The compound of claim 31 ,

where

wherein the wavy line across one non-cyclic bond indicates the bond that connects each Het group to A; where in a ring indicates the ring is an aromatic or heteroaromatic ring;

each W and W is independently a bond, C, CH, CH₂, CHR¹², CR¹², N, NH, NR¹², S, O, C=0, or C=S,

where each R is independently selected from H, halo, OR^x, SR^X, C0₂R^x, C(0)NR^x ₂, C(=0) R^x, CN, CF₃, OCF₃, N0₂, NR^X ₂, OCOR^x, S0₃R^x, P0₃R^x ₂, and CH(COOR^x)₂, where each R^x is H or C1-C4 alkyl or C1-C4 haloalkyl,

or R can be substituted or unsubstituted Ci_-8 alkyl, C₂_8 alkenyl, C₂_8 alkynyl;

and each R¹² is H or C1-C4 alkyl or C1-C4 haloalkyl,

or a pharmaceutically acceptable salt thereof.

33. The compound of claim 32, wherein Het is phenyl or pyridyl and is optionally substituted with up to three groups selected from C1-C4 alkyl, C1-C4 haloalkyl, halo, OR^x, SR^X, C0₂R^x, C(0)NR^X2, C(=0) R^x, CN, CF₃, OCF₃, N0₂, NR^X ₂, OCOR^x, S0₃R^x, P0₃R^x ₂, and CH(COOR^x)₂, where each R^x is H or C1-C4 alkyl or C1-C4 haloalkyl.

34. A compound of Formula Va and Formula

Formula Va Formula Vb wherein:

Z is S, SO, S0₂, Ci^R² NR^JR², or O;

A is a bond, C=0, CONH or NHCO;

Het is as defined for Formula I;

n is 1-4;

R¹, R² and R³ is independently selected from H, OH, Halo, NHR', NR'R',

OR', SR', COOR', C(=0)R', CN, CF₃, OCF₃, N0₂, OC(0)R', S0₃R', P0₃R'₂,

CR'(COOR')₂;

each R' is independently selected from H, halo, OR^x, SR^X, C0₂R^x,

C(0)NR^X2, C(=0) R^x, CN, CF₃, OCF₃, N0₂, NR^X ₂, OCOR^x, S0₃R^x, P0₃R^x ₂, and

CH(COOR^x)₂;

where each R^x is H or C1-C4 alkyl or C1-C4 haloalkyl;

or a pharmaceutically acceptable salt thereof.

35. The compound of claim 34,

where

wherein the wavy line across one non-cyclic bond indicates the bond that connects each Het group to A; where in a ring indicates the ring is an aromatic or heteroaromatic ring;

each W and W is independently a bond, C, CH, CH₂, CHR¹², CR¹², N, NH, NR¹², S, O, C=0, or C=S,

where each R is independently selected from H, halo, OR^x, SR^X, C0₂R^x, C(0)NR^x ₂, C(=0) R^x, CN, CF₃, OCF₃, N0₂, NR^X2, OCOR^x, S0₃R^x, P0₃R^x ₂, and CH(COOR^x)₂, where each R^x is H or C1-C4 alkyl or C1-C4 haloalkyl,

or R can be substituted or unsubstituted Ci_-8 alkyl, C2-8 alkenyl, C2-8 alkynyl;

and each R¹² is H or C1-C4 alkyl or C1-C4 haloalkyl,

or a pharmaceutically acceptable salt thereof.

36. The compound of claim 35, wherein Het is phenyl or pyridyl and is optionally substituted with up to three groups selected from C1-C4 alkyl, C1-C4 haloalkyl, halo, OR^x, SR^X, C0₂R^x, C(0)NR^X2, C(=0) R^x, CN, CF₃, OCF₃, N0₂, NR^X ₂, OCOR^x, S0₃R^x, P0₃R^x ₂, and CH(COOR^x)₂, where each R^x is H or C1-C4 alkyl or C1-C4 haloalkyl.

37. A pharmaceutical composition comprising a compound of any one of the claims 1-36 admixed with at least one pharmaceutically acceptable carrier or excipient.

38. The pharmaceutical composition of claim 37, which comprises at least one sterile pharmaceutically acceptable carrier or excipient.

39. The pharmaceutical composition of claim 37, which comprises at least two pharmaceutically acceptable carriers and/or excipients.

40. A compound according to any one of claims 1-36 for use in therapy.

41. The compound of claim 40, wherein the use in therapy is a use to treat cancer.

42. The compound of claim 41, wherein the cancer is selected from leukemia, lymphoma, lung cancer, colon cancer, CNS cancer, melanoma, ovarian cancer, renal cancer, prostate cancer, breast cancer, head and neck cancers, and pancreatic cancer.

43. A method to treat cancer, which comprises administering to a subject in need thereof an effective amount of a compound according to any one of claims 1-36.

44. The method of claim 43, wherein the cancer is selected from leukemia, lymphoma, lung cancer, colon cancer, CNS cancer, melanoma, ovarian cancer, renal cancer, prostate cancer, breast cancer, head and neck cancers, and pancreatic cancer.

45. Use of a compound according to any one of claims 1-36 for the manufacture of a medicament.

46. The use of claim 45, wherein the cancer is selected from leukemia, lymphoma, lung cancer, colon cancer, CNS cancer, melanoma, ovarian cancer, renal cancer, prostate cancer, breast cancer, head and neck cancers, and pancreatic cancer.