Tyrosine kinase inhibitors

BACKGROUND OF THE INVENTION

The present invention relates to compounds which inhibit, regulate and/or modulate tyrosine kinase signal transduction, compositions which contain these compounds, and methods of using them to treat tyrosine kinase-dependent diseases and conditions, such as angiogenesis, cancer, tumor growth, atherosclerosis, age related macular degeneration, diabetic retinopathy, inflammatory diseases, and the like in mammals.

Tyrosine kinases are a class of enzymes that catalyze the transfer of the terminal phosphate of adenosine triphosphate to tyrosine residues in protein substrates. Tyrosine kinases play critical roles in signal transduction for a number of cell functions via substrate phosphorylation. Though the exact mechanism of signal transduction is still unclear, tyrosine kinases have been shown to be important contributing factors in cell proliferation, carcinogenesis and cell differentiation.

Tyrosine kinases can be categorized as receptor type or non-receptor type. Receptor type tyrosine kinases have an extracellular, a transmembrane, and an intracellular portion, while non-receptor type tyrosine kinases are wholly intracellular.

The receptor type tyrosine kinases are comprised of a large number of transmembrane receptors with diverse biological activity. In fact, about twenty different subfamilies of receptor-type tyrosine kinases have been identified. One tyrosine kinase subfamily, designated the HER subfamily, is comprised of EGFR, HER2, HER3, and HER4. Ligands of this subfamily of receptors include epithileal growth factor, TGF-.alpha., amphiregulin, HB-EGF, betacellulin and heregulin. Another subfamily of these receptor-type tyrosine kinases is the insulin subfamily, which includes INS-R, IGF-IR, and IR-R. The PDGF subfamily includes the PDGF-.alpha. and .beta. receptors, CSFIR, c-kit and FLK-II. Then there is the FLK family which is comprised of the kinase insert domain receptor (KDR), fetal liver kinase-1 (FLK-1), fetal liver kinase-4 (FLK-4) and the fins-like tyrosine kinase-1 (flt-1). The PDGF and FLK families are usually considered together due to the similarities of the two groups. For a detailed discussion of the receptor-type tyrosine kinases, see Plowman et al., DN&P 7(6):334-339, 1994, which is hereby incorporated by reference.

The non-receptor type of tyrosine kinases is also comprised of numerous subfamilies, including Src, Frk, Btk, Csk, Abl, Zap70, Fes/Fps, Fak, Jak, Ack, and LIMK. Each of these subfamilies is further sub-divided into varying receptors. For example, the Src subfamily is one of the largest and includes Src, Yes, Fyn, Lyn, Lck, Blk, Hck, Fgr, and Yrk. The Src subfamily of enzymes has been linked to oncogenesis. For a more detailed discussion of the non-receptor type of tyrosine kinases, see Bolen Oncogene, 8:2025-2031 (1993), which is hereby incorporated by reference.

Both receptor type and non-receptor type tyrosine kinases are implicated in cellular signaling pathways leading to numerous pathogenic conditions, including cancer, psoriasis and hyperimmune responses.

Several receptor type tyrosine kinases, and the growth factors that bind thereto, have been suggested to play a role in angiogenesis, although some may promote angiogenesis indirectly (Mustonen and Alitalo, J. Cell Biol. 129:895-898, 1995). One such receptor type tyrosine kinase is fetal liver kinase 1 or FLK-1. The human analog of FLK-1 is the kinase insert domain-containing receptor KDR, which is also known as vascular endothelial cell growth factor receptor 2 or VEGFR-2, since it binds VEGF with high affinity. Finally, the murine version of this receptor has also been called NYK (Oelrichs et al., Oncogene 8(1): 11-15, 1993). VEGF and KDR are a ligand-receptor pair that play an important role in the proliferation of vascular endothelial cells, and the formation and sprouting of blood vessels, termed vasculogenesis and angiogenesis, respectively.

Angiogenesis is characterized by excessive activity of vascular endothelial growth factor (VEGF). VEGF is actually comprised of a family of ligands (Klagsburn and D'Amore, Cytokine &Growth Factor Reviews 7:259-270, 1996). VEGF binds the high affinity membrane-spanning tyrosine kinase receptor KDR and the related fms-like tyrosine kinase-1, also known as Flt-1 or vascular endothelial cell growth factor receptor 1 (VEGFR-1). Cell culture and gene knockout experiments indicate that each receptor contributes to different aspects of angiogenesis. KDR mediates the mitogenic function of VEGF whereas Flt-1 appears to modulate non-mitogenic functions such as those associated with cellular adhesion. Inhibiting KDR thus modulates the level of mitogenic VEGF activity. In fact, tumor growth has been shown to be susceptible to the antiangiogenic effects of VEGF receptor antagonists. (Kim et al., Nature 362, pp. 841-844, 1993).

Solid tumors can therefore be treated by tyrosine kinase inhibitors since these tumors depend on angiogenesis for the formation of the blood vessels necessary to support their growth. These solid tumors include histiocytic lymphoma, cancers of the brain, genitourinary tract, lymphatic system, stomach, larynx and lung, including lung adenocarcinoma and small cell lung cancer. Additional examples include cancers in which overexpression or activation of Raf-activating oncogenes (e.g., K-ras, erb-B) is observed. Such cancers include pancreatic and breast carcinoma. Accordingly, inhibitors of these tyrosine kinases are useful for the prevention and treatment of proliferative diseases dependent on these enzymes.

The angiogenic activity of VEGF is not limited to tumors. VEGF accounts for most of the angiogenic activity produced in or near the retina in diabetic retinopathy. This vascular growth in the retina leads to visual degeneration culminating in blindness. Ocular VEGF mRNA and protein are elevated by conditions such as retinal vein occlusion in primates and decreased pO.sub.2 levels in mice that lead to neovascularization. Intraocular injections of anti-VEGF monoclonal antibodies or VEGF receptor immunofusions inhibit ocular neovascularization in both primate and rodent models. Regardless of the cause of induction of VEGF in human diabetic retinopathy, inhibition of ocular VEGF is useful in treating the disease.

Expression of VEGF is also significantly increased in hypoxic regions of animal and human tumors adjacent to areas of necrosis. VEGF is also upregulated by the expression of the oncogenes ras, raf, src and mutant p53 (all of which are relevant to targeting cancer). Monoclonal anti-VEGF antibodies inhibit the growth of human tumors in nude mice. Although these same tumor cells continue to express VEGF in culture, the antibodies do not diminish their mitotic rate. Thus tumor-derived VEGF does not function as an autocrine mitogenic factor. Therefore, VEGF contributes to tumor growth in vivo by promoting angiogenesis through its paracrine vascular endothelial cell chemotactic and mitogenic activities. These monoclonal antibodies also inhibit the growth of typically less well vascularized human colon cancers in athymic mice and decrease the number of tumors arising from inoculated cells.

Viral expression of a VEGF-binding construct of Flk-1, Flt-1, the mouse KDR receptor homologue, truncated to eliminate the cytoplasmic tyrosine kinase domains but retaining a membrane anchor, virtually abolishes the growth of a transplantable glioblastoma in mice presumably by the dominant negative mechanism of heterodimer formation with membrane spanning endothelial cell VEGF receptors. Embryonic stem cells, which normally grow as solid tumors in nude mice, do not produce detectable tumors if both VEGF alleles are knocked out. Taken together, these data indicate the role of VEGF in the growth of solid tumors. Inhibition of KDR or Flt-1 is implicated in pathological angiogenesis, and these receptors are useful in the treatment of diseases in which angiogenesis is part of the overall pathology, e.g., inflammation, diabetic retinal vascularization, as well as various forms of cancer since tumor growth is known to be dependent on angiogenesis. (Weidner et al., N. Engl. J. Med., 324, pp. 1-8, 1991).

Indolinyl-isoquinolinone compounds which specifically inhibit, regulate and/or modulate the signal transduction of tyrosine kinases have been previously reported, see WO 01/29025, published 26 Apr. 2001. The identification of compounds with improved pharmaceutical properties, however, remains desireable and is an object of this invention.

SUMMARY OF THE INVENTION

The present invention relates to compounds that are capable of inhibiting, modulating and/or regulating signal transduction of both receptor-type and non-receptor type tyrosine kinases. One embodiment of the present invention is illustrated by a compound of Formula I, and the pharmaceutically acceptable salts and stereoisomers thereof: ##STR1##

DETAILED DESCRIPTION OF THE INVENTION

The compounds of this invention are useful in the inhibition of kinases and are illustrated by a compound of Formula I: ##STR2##

or a pharmaceutically acceptable salt or stereoisomer thereof, wherein a is 0 or 1; b is 0 or 1; m is 0, 1, or 2; t is 1 or 2; R.sup.1 and R.sup.5 are independently selected from:

1) H,

2) (C.dbd.O).sub.a O.sub.b C.sub.1 -C.sub.10 alkyl,

3) (C.dbd.O).sub.a O.sub.b aryl,

4) (C.dbd.O).sub.a O.sub.b C.sub.2 -C.sub.10 alkenyl,

5) (C.dbd.O).sub.a O.sub.b C.sub.2 -C.sub.10 alkynyl,

6) CO.sub.2 H,

7) halo,

8) OH,

9) O.sub.b C.sub.1 -C.sub.6 perfluoroalkyl,

10) (C.dbd.O).sub.a NR.sup.7 R.sup.8,

11) CN,

12) (C.dbd.O).sub.a O.sub.b C.sub.3 -C.sub.8 cycloalkyl, and

13) (C.dbd.O).sub.a O.sub.b heterocyclyl, said alkyl, aryl, alkenyl, alkynyl, cycloalkyl, and heterocyclyl is optionally substituted with one or more substituents selected from R.sup.6 ; R.sup.2 and R.sup.3 are independently selected from:

1) H,

2) (C.dbd.O)O.sub.a C.sub.1 -C.sub.6 alkyl,

3) (C.dbd.O)O.sub.a aryl,

4) C.sub.1 -C.sub.6 alkyl,

5) SO.sub.2 R.sup.a, and

6) aryl; R.sup.4a or R.sup.4b is H and the other is selected from:

1) (C.dbd.O).sub.a O.sub.b C.sub.1 -C.sub.10 alkyl,

2) (C.dbd.O).sub.a O.sub.b aryl,

3) (C.dbd.O).sub.a O.sub.b C.sub.2 -C.sub.10 alkenyl,

4) (C.dbd.O).sub.a O.sub.b C.sub.2 -C.sub.10 alkynyl,

5) CO.sub.2 H,

6) halo,

7) OH,

8) O.sub.b C.sub.1 -C.sub.6 perfluoroalkyl,

9) (C.dbd.O).sub.a NR.sup.7 R.sup.8,

10) CN,

11) (C.dbd.O).sub.a O.sub.b C.sub.3 -C.sub.8 cycloalkyl, and

12) (C.dbd.O).sub.a O.sub.b heterocyclyl, said alkyl, aryl, alkenyl, alkynyl, cycloalkyl, and heterocyclyl is optionally substituted with one or more substituents selected from R.sup.6 ; R.sup.6 is:

1) (C.dbd.O).sub.a O.sub.b C.sub.1 -C.sub.10 alkyl,

2) (C.dbd.O).sub.a O.sub.b aryl,

3) C.sub.2 -C.sub.10 alkenyl,

4) C.sub.2 -C.sub.10 alkynyl,

5) (C.dbd.O).sub.a O.sub.b heterocyclyl,

6) CO.sub.2 H,

7) halo,

8) CN,

9) OH,

10) O.sub.b C.sub.1 -C.sub.6 perfluoroalkyl, or

11) O.sub.a (C.dbd.O).sub.b NR.sup.7 R.sup.8,

12) oxo,

13) CHO,

14) (N.dbd.O)R.sup.7 R.sup.8, and

15) (C.dbd.O).sub.a O.sub.b C.sub.3 -C.sub.8 cycloalkyl, said alkyl, aryl, alkenyl, alkynyl, heterocyclyl, and cycloalkyl optionally substituted with one or more substituents selected from R.sup.6a ; R.sup.6a is selected from:

1) (C.dbd.O).sub.r O.sub.s (C.sub.1 -C.sub.10)alkyl, wherein r and s are independently 0 or 1,

2) O.sub.r (C.sub.1 -C.sub.3)perfluoroalkyl, wherein r is 0 or 1,

3) (C.sub.0 -C.sub.6)alkylene-S(O).sub.m R.sup.a, wherein m is 0, 1, or 2,

4) SO.sub.2 N(R.sup.b).sub.2,

5) oxo,

6) OH,

7) halo,

8) CN,

9) (C.sub.2 -C.sub.10)alkenyl,

10) (C.sub.2 -C.sub.10)alkynyl,

11) (C.sub.3 -C.sub.6)cycloalkyl,

12) (C.sub.0 -C.sub.6)alkylene-aryl,

13) (C.sub.0 -C.sub.6)alkylene-heterocyclyl,

14) (C.sub.0 -C.sub.6)alkylene-N(R.sup.b).sub.2,

15) C(O)R.sup.a,

16) (C.sub.0 -C.sub.6)alkylene-CO.sub.2 R.sup.a,

17) C(O)H, and

18) (C.sub.0 -C.sub.6)alkylene-CO.sub.2 H, said alkyl, alkenyl, alkynyl, cycloalkyl, aryl, and heterocyclyl is optionally substituted with up to three substituents selected from R.sup.b, OH, (C.sub.1 -C.sub.6)alkoxy, halogen, CO.sub.2 H, CN, O(C.dbd.O)C.sub.1 -C.sub.6 alkyl, oxo, and N(R.sup.b).sub.2 ; R.sup.7 and R.sup.8 are independently selected from:

1) H,

2) (C.dbd.O)O.sub.b C.sub.1 -C.sub.10 alkyl,

3) (C.dbd.O)O.sub.b C.sub.3 -C.sub.8 cycloalkyl,

4) (C.dbd.O)O.sub.b aryl,

5) (C.dbd.O)O.sub.b heterocyclyl,

6) C.sub.1 -C.sub.10 alkyl,

7) aryl,

8) C.sub.2 -C.sub.10 alkenyl,

9) C.sub.2 -C.sub.10 alkynyl,

10) heterocyclyl,

11) C.sub.3 -C.sub.8 cycloalkyl,

12) SO.sub.2 R.sup.a,

13) (C.dbd.O)N(R.sup.b).sub.2, said alkyl, cycloalkyl, aryl, heterocylyl, alkenyl, and alkynyl is optionally substituted with one or more substituents selected from R.sup.6a, or R.sup.7 and R.sup.8 can be taken together with the nitrogen to which they are attached to form a monocyclic or bicyclic heterocycle with 5-7 members in each ring and optionally containing, in addition to the nitrogen, one or two additional heteroatoms selected from N, O and S, said monocylcic or bicyclic heterocycle optionally substituted with one or more substituents selected from R.sup.6a ; R.sup.a is (C.sub.1 -C.sub.6)alkyl, (C.sub.3 -C.sub.6)cycloalkyl, aryl, or heterocyclyl; and R.sup.b is H, (C.sub.1 -C.sub.6)alkyl, aryl, heterocyclyl, (C.sub.3 -C.sub.6)cycloalkyl, (C.dbd.O)OC.sub.1 -C.sub.6 alkyl, (C.dbd.O)C.sub.1 -C.sub.6 alkyl or S(O).sub.2 R.sup.a.

A second embodiment is a compound as described above, wherein t is 1; R.sup.1 and R.sup.5 are independently selected from:

1) H,

2) (C.dbd.O).sub.a O.sub.b C.sub.1 -C.sub.6 alkyl,

3) (C.dbd.O).sub.a O.sub.b aryl,

4) (C.dbd.O).sub.a O.sub.b C.sub.2 -C.sub.6 alkenyl,

5) (C.dbd.O).sub.a O.sub.b C.sub.2 -C.sub.6 alkynyl,

6) CO.sub.2 H,

7) halo,

8) OH,

9) O.sub.b C.sub.1 -C.sub.3 perfluoroalkyl,

10) (C.dbd.O).sub.a NR.sup.7 R.sup.8,

11) CN,

12) (C.dbd.O).sub.a O.sub.b C.sub.3 -C.sub.6 cycloalkyl, and

13) (C.dbd.O).sub.a O.sub.b heterocyclyl, said alkyl, aryl, alkenyl, alkynyl, cycloalkyl, and heterocyclyl is optionally substituted with one or more substituents selected from R.sup.6 ; R.sup.4a or R.sup.4b is H and the other is selected from:

1) (C.dbd.O).sub.a O.sub.b C.sub.1 -C.sub.6 alkyl,

2) (C.dbd.O).sub.a O.sub.b aryl,

3) (C.dbd.O).sub.a O.sub.b C.sub.2 -C.sub.6 alkenyl,

4) (C.dbd.O).sub.a O.sub.b C.sub.2 -C.sub.6 alkynyl,

5) CO.sub.2 H,

6) halo,

7) OH,

8) O.sub.b C.sub.1 -C.sub.3 perfluoroalkyl,

9) (C.dbd.O).sub.a NR.sup.7 R.sup.8,

10) CN,

11) (C.dbd.O).sub.a O.sub.b C.sub.3 -C.sub.6 cycloalkyl, and

12) (C.dbd.O).sub.a O.sub.b heterocyclyl, said alkyl, aryl, alkenyl, alkynyl, cycloalkyl, and heterocyclyl is optionally substituted with one or more substituents selected from R.sup.6 ; R.sup.6 is:

1) (C.dbd.O).sub.a O.sub.b C.sub.1 -C.sub.6 alkyl,

2) (C.dbd.O).sub.a O.sub.b aryl,

3) C.sub.2 -C.sub.6 alkenyl,

4) C.sub.2 -C.sub.6 alkynyl,

5) (C.dbd.O).sub.a O.sub.b heterocyclyl,

6) CO.sub.2 H,

7) halo,

8) CN,

9) OH,

10) O.sub.b C.sub.1 -C.sub.3 perfluoroalkyl, or

11) O.sub.a (C.dbd.O).sub.b NR.sup.7 R.sup.8,

12) oxo,

13) CHO,

14) (N.dbd.O)R.sup.7 R.sup.8, and

15) (C.dbd.O).sub.a O.sub.b C.sub.3 -C.sub.6 cycloalkyl, said alkyl, aryl, alkenyl, alkynyl, heterocyclyl, and cycloalkyl is optionally substituted with one or more substituents selected from R.sup.6a ; R.sup.6a is selected from:

1) (C.dbd.O).sub.r O.sub.s (C.sub.1 -C.sub.6)alkyl, wherein r and s are independently 0 or 1,

2) O.sub.r (C.sub.1 -C.sub.3)perfluoroalkyl, wherein r is 0 or 1,

3) (C.sub.0 -C.sub.6)alkylene-S(O).sub.m R.sup.a, wherein m is 0, 1, or 2,

4) SO.sub.2 N(R.sup.b).sub.2,

5) oxo,

6) OH,

7) halo,

8) CN,

9) (C.sub.2 -C.sub.6)alkenyl,

10) (C.sub.2 -C.sub.6)alkynyl,

11) (C.sub.3 -C.sub.6)cycloalkyl,

12) (C.sub.0 -C.sub.6)alkylene-aryl,

13) (C.sub.0 -C.sub.6)alkylene-heterocyclyl,

14) (C.sub.0 -C.sub.6)alkylene-N(R.sup.b).sub.2,

15) C(O)R.sup.a,

16) (C.sub.0 -C.sub.6)alkylene-CO.sub.2 R.sup.a,

17) C(O)H, and

18) (C.sub.0 -C.sub.6)alkylene-CO.sub.2 H, said alkyl, alkenyl, alkynyl, cycloalkyl, aryl, and heterocyclyl is optionally substituted with up to three substituents selected from R.sup.b, OH, (C.sub.1 -C.sub.6)alkoxy, halogen, CO.sub.2 H, CN, O(C.dbd.O)C.sub.1 -C.sub.6 alkyl, oxo, and N(R.sup.b).sub.2 ; and R.sup.7 and R.sup.8 are independently selected from:

1) H,

2) (C.dbd.O)O.sub.b C.sub.1 -C.sub.6 alkyl,

3) (C.dbd.O)O.sub.b C.sub.3 -C.sub.6 cycloalkyl,

4) (C.dbd.O)O.sub.b aryl,

5) (C.dbd.O)O.sub.b heterocyclyl,

6) C.sub.1 -C.sub.6 alkyl,

7) aryl,

8) C.sub.2 -C.sub.6 alkenyl,

9) C.sub.2 -C.sub.6 alkynyl,

10) heterocyclyl,

11) C.sub.3 -C.sub.6 cycloalkyl,

12) SO.sub.2 R.sup.a,

13) (C.dbd.O)N(R.sup.b).sub.2, said alkyl, cycloalkyl, aryl, heterocylyl, alkenyl, and alkynyl is optionally substituted with one or more substituents selected from R.sup.6a, or R.sup.7 and R.sup.8 can be taken together with the nitrogen to which they are attached to form a monocyclic or bicyclic heterocycle with 5-7 members in each ring and optionally containing, in addition to the nitrogen, one or two additional heteroatoms selected from N, O and S, said monocylcic or bicyclic heterocycle optionally substituted with one or more substituents selected from R.sup.6a.

A third embodiment of the present invention is illustrated by a compound as described above, wherein R.sup.2 and R.sup.3 are H; and R.sup.5 is H or F.

And yet another embodiment is a compound as described above, wherein R.sup.1 is H.

A further embodiment is a compound as described above, wherein R.sup.4a or R.sup.4b is H and the other is C.sub.1 -C.sub.6 alkyleneNR.sup.7 R.sup.8, said alkylene optionally substituted with oxo.

A preferred ambodiment is a compound selected from: 3-{6-[(4-methylpiperazin-1-yl)carbonyl]-1H-indol-2-yl}quinolin-2(1H)-one; N-methyl-2-(2-oxo-1,2-dihydroquinolin-3-yl)-N-pyrrolidin-3-yl-1H-indole-6-c arboxamide; 3-{4-[(4-methylpiperazin-1-yl)carbonyl]-1H-indol-2-yl}quinolin-2(1H)-one; N-[2-(dimethylamino)ethyl]-2-(2-oxo-1,2-dihydroquinolin-3-yl)-1H-indole-4-c arboxamide; and N-methyl-2-(2-oxo-1,2-dihydroquinolin-3-yl)-N-pyrrolidin-3-yl-1H-indole-4-c arboxamide; or a pharmaceutically acceptable salt or stereoisomer thereof.

Another preferred ambodiment is a compound selected from: 3-(6-{[4-(methylsulfonyl)piperazin-1-yl]methyl}-1H-indol-2-yl)quinolin-2(1H )-one; 4-[2-(2-Oxo-1,2-dihydro-quinolin-3-yl)-1H-indol-6-ylmethyl]-piperazine-1-ca rboxylic acid methylamide; 3-{4-[2-(2-Oxo-1,2-dihydro-quinolin-3-yl)-1H-indol-6-ylmethyl]-piperazin-1- yl}-butyric acid; 3-[4-(4-Methanesulfonyl-piperazin-1-ylmethyl)-1H-indol-2-yl]-1H-quinolin-2- one; and 3-{4-[2-(2-Oxo-1,2-dihydro-quinolin-3-yl)-1H-indol-4-ylmethyl]-piperazin-1- yl}-butyric acid, or a pharmaceutically acceptable salt or stereoisomer thereof.

And yet another preferred ambodiment is a compound selected from: 3-[3-fluoro-6-(4-methanesulfonyl-piperazin-1-ylmethyl)-1H-indol-2-yl]-1H-qu inolin-2-one; 4-[2-(3-fluoro-2-oxo-1,2-dihydro-quinolin-3-yl)-1H-indol-6-ylmethyl]-pipera zine-1-carboxylic acid methylamide; 3-{4-[3-fluoro-2-(2-oxo-1,2-dihydro-quinolin-3-yl)-1H-indol-6-ylmethyl]-pip erazin-1-yl}-butyric acid; 3-[3-fluoro-4-(4-methanesulfonyl-piperazin-1-ylmethyl)-1H-indol-2-yl]-1H-qu inolin-2-one; and 3-{4-[3-fluoro-2-(2-oxo-1,2-dihydro-quinolin-3-yl)-1H-indol-4-ylmethyl]-pip erazin-1-yl}-butyric acid; or a pharmaceutically acceptable salt or stereoisomer thereof.

Also included within the scope of the present invention is a pharmaceutical composition which is comprised of a compound of Formula I as described above and a pharmaceutically acceptable carrier. The invention is also contemplated to encompass a pharmaceutical composition which is comprised of a pharmaceutically acceptable carrier and any of the compounds specifically disclosed in the present application. These and other aspects of the invention will be apparent from the teachings contained herein.

Utilities

The instantly disclosed compounds are inhibitors of tyrosine kinase and are therefore useful to treat or prevent tyrosine kinase-dependent diseases or conditions in mammals.

"Tyrosine kinase-dependent diseases or conditions" refers to pathologic conditions that depend on the activity of one or more tyrosine kinases. Tyrosine kinases either directly or indirectly participate in the signal transduction pathways of a variety of cellular activities including proliferation, adhesion and migration, and differentiation. Diseases associated with tyrosine kinase activities include the proliferation of tumor cells, the pathologic neovascularization that supports solid tumor growth, ocular neovascularization (diabetic retinopathy, age-related macular degeneration, and the like) and inflammation (psoriasis, rheumatoid arthritis, and the like). In treating such conditions with the instantly claimed compounds, the required therapeutic amount will vary according to the specific disease and is readily ascertainable by those skilled in the art. Although both treatment and prevention are contemplated by the scope of the invention, the treatment of these conditions is the preferred use.

The present invention encompasses a method of treating or preventing cancer in a mammal in need of such treatment which is comprised of administering to said mammal a therapeutically effective amount of a compound of Formula I. Preferred cancers for treatment are selected from cancers of the brain, genitourinary tract, lymphatic system, stomach, larynx and lung. Another set of preferred forms of cancer are histiocytic lymphoma, lung adenocarcinoma, small cell lung cancers, pancreatic cancer, glioblastomas and breast carcinoma. The utility of angiogenesis inhibitors in the treatment of cancer is known in the literature, see J. Rak et al. Cancer Research, 55:4575-4580, 1995, for example. The role of angiogenesis in cancer has been shown in numerous types of cancer and tissues: breast carcinoma (G. Gasparini and A. L. harris, J. Clin. Oncol., 1995, 13:765-782; M. Toi et al., Japan, J. Cancer Res., 1994, 85:1045-1049); bladder carcinomas (A. J. Dickinson et al., Br. J. Urol., 1994, 74:762-766); colon carcinomas (L. M. Ellis et al., Surgery, 1996, 120(5):871-878); and oral cavity tumors (J. K. Williams et al., Am. J. Surg., 1994, 168:373-380).

Tumors which have undergone neovascularization show an increased potential for metastasis. VEGF released from cancer cells enhances metastasis possibly by increasing extravasation at points of adhesion to vascular endothelium. (A. Amirkhosravi et al., Platelets, 10:285-292 (1999).) In fact, angiogenesis is essential for tumor growth and metastasis. (S. P. Gunningham, et al., Can. Research, 61: 3206-3211 (2001).) The angiogenesis inhibitors disclosed in the present application are therefore also useful to prevent or decrease tumor cell metastasis. Such a use is also contemplated to be within the scope of the present invention.

Further included within the scope of the invention is a method of treating or preventing a disease in which angiogenesis is implicated, which is comprised of administering to a mammal in need of such treatment a therapeutically effective amount of a compound of Formula I. Ocular neovascular diseases are an example of conditions where much of the resulting tissue damage can be attributed to aberrant infiltration of blood vessels in the eye (see WO 00/30651, published 2 Jun. 2000). The undesireable infiltration can be triggered by ischemic retinopathy, such as that resulting from diabetic retinopathy, retinopathy of prematurity, retinal vein occlusions, etc., or by degenerative diseases, such as the choroidal neovascularization obeserved in age-related macular degeneration. Inhibiting the growth of blood vessels by administration of the present compounds would therefore prevent the infiltration of blood vessels and prevent or treat diseases where angiogenesis is implicated, such as ocular diseases like retinal vascularization, diabetic retinopathy, age-related macular degeneration, and the like.

Also included within the scope of the present invention is a method of treating or preventing inflammatory diseases which comprises administering to a mammal in need of such treatment a therapeutically effective amount of a compound of Formual I. Examples of such inflammatory diseases are rheumatoid arthritis, psoriasis, contact dermatitis, delayed hypersensitivity reactions, and the like. (A. Giatromanolaki et al., J. Pathol. 2001; 194:101-108.) For the role of VEGF in skin angiogenesis, see Michael Detmar, J. Dermatological Sci., 24 Suppl. 1, S78-S84 (2000).

Also included within the scope of the present invention is a method of treating or preventing bone associated pathologies selected from osteosarcoma, osteoarthritis, and rickets, also known as oncogenic osteomalacia. (Hasegawa et al., Skeletal Radiol., 28, pp. 41-45, 1999; Gerber et al., Nature Medicine, Vol. 5, No. 6, pp. 623-628, June 1999.) And since VEGF directly promotes osteoclastic bone resorption through KDR/Flk-1 expressed in mature osteoclasts (FEBS Let. 473:161-164 (2000); Endocrinology, 141:1667 (2000)), the instant compounds are also useful to treat and prevent conditions related to bone resorption, such as osteoporosis and Paget's disease.

A method of treating or preventing preeclampsia is also within the which comprises administering a therapeutically effective amount of a compound of Formula I. Studies have shown that the action of VEGF on the Flt-1 receptor is pivotal in the pathogenesis of preeclampsia (Laboratory Investigation 79:1101-1111 (September 1999).) Vessels of pregnant women incubated with VEGF exhibit a reduction in endothelium-dependent relaxation similar to that induced by plasma from women with preeclampsia. In the presence of an anti-Flt-1 receptor antibody, however, neither VEGF or plasma from women with preeclampsia reduced the endothelium-dependent relaxation. Therefore the claimed compounds serve to treat preeclampsia via their action on the tyrosine kinase domain of the Flt-1 receptor.

Also within the scope of the invention is a method of reducing or preventing tissue damage following a cerebral ischemic event which comprises administering a therapeutically effective amount of a compound of Formula I. The claimed compounds can also be used to reduce or prevent tissue damage which occurs after cerebral ischemic events, such as stroke, by reducing cerebral edema, tissue damage, and reperfusion injury following ischemia. (Drug News Perspect 11:265-270 (1998); J. Clin. Invest. 104:1613-1620 (1999); Nature Med 7:222-227 (2001)).

The instant compounds can also be used to prevent or treat tissue damage during bacterial meningitis, such as tuberculous meningitis. Matsuyama et al., J. Neurol. Sci. 186:75-79 (2001)). The instant invention therefore encompasses a method of treating or preventing tissue damage due to bacterial meningitis which comprises administering a therapeutically effective amount of a compound of Formula 1. Studies have shown that VEGF is secreted by inflammatory cells during bacterial meningitis and that VEGF contributes to blood-brain barrier disruption. (van der Flier et al., J. Infectious Diseases, 183:149-153 (2001)). The claimed compounds can inhibit VEGF-induced vascular permeability and therefore serve to prevent or treat blood-brain barrier disruption associated with bacterial meningitis.

The present invention further encompasses a method to treat or prevent endometrioses comprised of administering a therapeutically effective amount of a compound of Formula I. An increase in VEGF expression and angiogenesis is associated with the progression of endometriosis (Stephen K. Smith, Trends in Endocrinology & Metabolism, Vol. 12, No. 4, May/June 2001). Inhibition of VEGF by the current compounds would therefore inhibit angiogenesis and treat endometriosis.

The compounds of this invention may be administered to mammals, preferably humans, either alone or, preferably, in combination with pharmaceutically acceptable carriers or diluents, optionally with known adjuvants, such as alum, in a pharmaceutical composition, according to standard pharmaceutical practice. The compounds can be administered orally or parenterally, including the intravenous, intramuscular, intraperitoneal, subcutaneous, rectal and topical routes of administration.

For oral use of a chemotherapeutic compound according to this invention, the selected compound may be administered, for example, in the form of tablets or capsules, or as an aqueous solution or suspension. In the case of tablets for oral use, carriers which are commonly used include lactose and corn starch, and lubricating agents, such as magnesium stearate, are commonly added. For oral administration in capsule form, useful diluents include lactose and dried corn starch. When aqueous suspensions are required for oral use, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening and/or flavoring agents may be added. For intramuscular, intraperitoneal, subcutaneous and intravenous use, sterile solutions of the active ingredient are usually prepared, and the pH of the solutions should be suitably adjusted and buffered. For intravenous use, the total concentration of solutes should be controlled in order to render the preparation isotonic.

The instant compounds are also useful in combination with known anti-cancer agents. Such known anti-cancer agents include the following: estrogen receptor modulators, androgen receptor modulators, retinoid receptor modulators, cytotoxic agents, antiproliferative agents, prenyl-protein transferase inhibitors, HMG-CoA reductase inhibitors, HIV protease inhibitors, reverse transcriptase inhibitors, and other angiogenesis inhibitors. The instant compounds are particularly useful when coadminsitered with radiation therapy. The synergistic effects of inhibiting VEGF in combination with radiation therapy have been described in the art (see WO 00/61186).

"Estrogen receptor modulators" refers to compounds which interfere or inhibit the binding of estrogen to the receptor, regardless of mechanism. Examples of estrogen receptor modulators include, but are not limited to, tamoxifen, raloxifene, idoxifene, LY353381, LY117081, toremifene, fulvestrant, 4-[7-(2,2-dimethyl-1-oxopropoxy-4-methyl-2-[4-[2-(1-piperidinyl)ethoxy]phe nyl]-2H-1-benzopyran-3-yl]-phenyl-2,2-dimethylpropanoate, 4,4'-dihydroxybenzophenone-2,4-dinitrophenyl-hydrazone, and SH646.

"Androgen receptor modulators" refers to compounds which interfere or inhibit the binding of androgens to the receptor, regardless of mechanism. Examples of androgen receptor modulators include finasteride and other 5.alpha.-reductase inhibitors, nilutamide, flutamide, bicalutamide, liarozole, and abiraterone acetate.

"Retinoid receptor modulators" refers to compounds which interfere or inhibit the binding of retinoids to the receptor, regardless of mechanism. Examples of such retinoid receptor modulators include bexarotene, tretinoin, 13-cis-retinoic acid, 9-cis-retinoic acid, .alpha.-difluoromethylornithine, ILX23-7553, trans-N-(4'-hydroxyphenyl)retinamide, and N-4-carboxyphenyl retinamide.

"Cytotoxic agents" refer to compounds which cause cell death primarily by interfering directly with the cell's functioning or inhibit or interfere, with cell myosis, including alkylating agents, tumor necrosis factors, intercalators, microtubulin inhibitors, and topoisomerase inhibitors.

Examples of cytotoxic agents include, but are not limited to, tirapazimine, sertenef, cachectin, ifosfamide, tasonermin, lonidamine, carboplatin, altretamine, prednimustine, dibromodulcitol, ranimustine, fotemustine, nedaplatin, oxaliplatin, temozolomide, heptaplatin, estramustine, improsulfan tosilate, trofosfamide, nimustine, dibrospidium chloride, pumitepa, lobaplatin, satraplatin, profiromycin, cisplatin, irofulven, dexifosfamide, cis-aminedichloro(2-methyl-pyridine)platinum, benzylguanine, glufosfamide, GPX100, (trans, trans, trans)-bis-mu-(hexane-1,6-diamine)-mu-[diamine-platinum(II)]bis[diamine(ch loro)platinum (II)]tetrachloride, diarizidinylspermine, arsenic trioxide, 1-(11-dodecylamino-10-hydroxyundecyl)-3,7-dimethylxanthin, zorubicin, idarubicin, daunorubicin, bisantrene, mitoxantrone, pirarubicin, pinafide, valrubicin, amrubicin, antineoplaston, 3'-deamino-3'-morpholino-13-deoxo-10-hydroxycarminomycin, annamycin, galarubicin, elinafide, MN10755, and 4-demethoxy-3-deamino-3-aziridinyl-4-methylsulphonyl-daunorubicin (see WO 00/50032).

Examples of microtubulin inhibitors include paclitaxel, vindesine sulfate, 3',4'-didehydro-4'-deoxy-8'-norvincaleukoblastine, docetaxol, rhizoxin, dolastatin, mivobulin isethionate, auristatin, cemadotin, RPR109881, BMS184476, vinflunine, cryptophycin, 2,3,4,5,6-pentafluoro-N-(3-fluoro-4-methoxyphenyl) benzene sulfonamide, anhydrovinblastine, N,N-dimethyl-L-valyl-L-valyl-N-methyl-L-valyl-L-prolyl-L-proline-t-butylam ide, TDX258, and BMS188797.

Some examples of topoisomerase inhibitors are topotecan, hycaptamine, irinotecan, rubitecan, 6-ethoxypropionyl-3',4'-O-exo-benzylidene-chartreusin, 9-methoxy-N,N-dimethyl-5-nitropyrazolo[3,4,5-kl]acridine-2-(6H) propanamine, 1-amino-9-ethyl-5-fluoro-2,3-dihydro-9-hydroxy-4-methyl-1H, 12H-benzo[de]pyrano[3',4': b,7]indolizino[1,2b]quinoline-10,13(9H,15H)dione, lurtotecan, 7-[2-(N-isopropylamino)ethyl]-(20S)camptothecin, BNP1350, BNPI1100, BN80915, BN80942, etoposide phosphate, teniposide, sobuzoxane, 2'-dimethylamino-2'-deoxy-etoposide, GL331, N-[2-(dimethylamino)ethyl]-9-hydroxy-5,6-dimethyl-6H-pyrido[4,3-b]carbazol e-1-carboxamide, asulacrine, (5a, 5aB, 8aa,9b)-9-[2-[N-[2-(dimethylamino)ethyl]-N-methylamino]ethyl]-5-[4-hydroxy -3,5-dimethoxyphenyl]-5,5a,6,8,8a,9-hexohydrofuro(3',4': 6,7)naphtho(2,3-d)-1,3-dioxol-6-one, 2,3-(methylenedioxy)-5-methyl-7-hydroxy-8-methoxybenzo[c]-phenanthridinium , 6,9-bis[(2-aminoethyl)amino]benzo[g]isoguinoline-5,10-dione, 5-(3-aminopropylamino)-7,10-dihydroxy-2-(2-hydroxyethylaminomethyl)-6H-pyr azolo[4,5,1-de]acridin-6-one, N-[1-[2(diethylamino)ethylamino]-7-methoxy-9-oxo-9H-thioxanthen-4-ylmethyl ]formamide, N-(2-(dimethylamino)ethyl)acridine-4-carboxamide, 6-[[2-(dimethylamino)ethyl]amino]-3-hydroxy-7H-indeno[2,1-c]quinolin-7-one , and dimesna.

"Antiproliferative agents" includes antisense RNA and DNA oligonucleotides such as G3139, ODN698, RVASKRAS, GEM231, and INX3001, and antimetabolites such as enocitabine, carmofur, tegafur, pentostatin, doxifluridine, trimetrexate, fludarabine, capecitabine, galocitabine, cytarabine ocfosfate, fosteabine sodium hydrate, raltitrexed, paltitrexid, ernitefur, tiazofurin, decitabine, nolatrexed, pemetrexed, nelzarabine, 2'-deoxy-2'-methylidenecytidine, 2'-fluoromethylene-2'-deoxycytidine, N-[5-(2,3-dihydro-benzofuryl)sulfonyl]-N'-(3,4-dichlorophenyl) urea, N6-[4-deoxy-4-[N2-[2(E),4(E)-tetradecadienoyl]glycylamino]-L-glycero-B-L-m anno-heptopyranosyl]adenine, aplidine, ecteinascidin, troxacitabine, 4-[2-amino-4-oxo-4,6,7,8-tetrahydro-3H-pyrimidino[5,4-b][1,4]thiazin-6-yl- (S)-ethyl]-2,5-thienoyl-L-glutamic acid, aminopterin, 5-flurouracil, alanosine, 11-acetyl-8-(carbamoyloxymethyl)-4-formyl-6-methoxy-14-oxa-1,11-diazatetra cyclo(7.4.1.0.0)-tetradeca-2,4,6-trien-9-yl acetic acid ester, swainsonine, lometrexol, dexrazoxane, methioninase, 2'-cyano-2'-deoxy-N4-palmitoyl-1-B-D-arabino furanosyl cytosine, and 3-aminopyridine-2-carboxaldehyde thiosemicarbazone. "Antiproliferative agents" also includes monoclonal antibodies to growth factors, other than those listed under "angiogenesis inhibitors", such as trastuzumab, and tumor suppressor genes, such as p53, which can be delivered via recombinant virus-mediated gene transfer (see U.S. Pat. No. 6,069,134, for example).

"HMG-CoA reductase inhibitors" refers to inhibitors of 3-hydroxy-3-methylglutaryl-CoA reductase. Compounds which have inhibitory activity for HMG-CoA reductase can be readily identified by using assays well-known in the art. For example, see the assays described or cited in U.S. Pat. No. 4,231,938 at col. 6, and WO 84/02131 at pp. 30-33. The terms "HMG-CoA reductase inhibitor" and "inhibitor of HMG-CoA reductase" have the same meaning when used herein.

Examples of HMG-CoA reductase inhibitors that may be used include but are not limited to lovastatin (MEVACOR.RTM.; see U.S. Pat. Nos. 4,231,938, 4,294,926 and 4,319,039), simvastatin (ZOCOR.RTM.; see U.S. Pat. Nos. 4,444,784, 4,820,850 and 4,916,239), pravastatin (PRAVACHOL.RTM.; see U.S. Pat. Nos. 4,346,227, 4,537,859, 4,410,629, 5,030,447 and 5,180,589), fluvastatin (LESCOL.RTM.; see U.S. Pat. Nos. 5,354,772, 4,911,165, 4,929,437, 5,189,164, 5,118,853, 5,290,946 and 5,356,896), atorvastatin (LIPITOR.RTM.; see U.S. Pat. Nos. 5,273,995, 4,681,893, 5,489,691 and 5,342,952) and cerivastatin (also known as rivastatin and BAYCHOL.RTM.; see U.S. Pat. No. 5,177,080). The structural formulas of these and additional HMG-CoA reductase inhibitors that may be used in the instant methods are described at page 87 of M. Yalpani, "Cholesterol Lowering Drugs", Chemistry & Industry, pp. 85-89 (5 Feb. 1996) and U.S. Pat. Nos. 4,782,084 and 4,885,314. The term HMG-CoA reductase inhibitor as used herein includes all pharmaceutically acceptable lactone and open-acid forms (i.e., where the lactone ring is opened to form the free acid) as well as salt and ester forms of compounds which have HMG-CoA reductase inhibitory activity, and therefor the use of such salts, esters, open-acid and lactone forms is included within the scope of this invention. An illustration of the lactone portion and its corresponding open-acid form is shown below as structures I and II. ##STR3##

In HMG-CoA reductase inhibitors where an open-acid form can exist, salt and ester forms may preferably be formed from the open-acid, and all such forms are included within the meaning of the term "HMG-CoA reductase inhibitor" as used herein. Preferably, the HMG-CoA reductase inhibitor is selected from lovastatin and simvastatin, and most preferably simvastatin. Herein, the term "pharmaceutically acceptable salts" with respect to the HMG-CoA reductase inhibitor shall mean non-toxic salts of the compounds employed in this invention which are generally prepared by reacting the free acid with a suitable organic or inorganic base, particularly those formed from cations such as sodium, potassium, aluminum, calcium, lithium, magnesium, zinc and tetramethylammonium, as well as those salts formed from amines such as ammonia, ethylenediamine, N-methylglucamine, lysine, arginine, ornithine, choline, N,N'-dibenzylethylenediamine, chloroprocaine, diethanolamine, procaine, N-benzylphenethylamine, 1-p-chlorobenzyl-2-pyrrolidine-1'-yl-methylbenzimidazole, diethylamine, piperazine, and tris(hydroxymethyl)aminomethane. Further examples of salt forms of HMG-CoA reductase inhibitors may include, but are not limited to, acetate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, calcium edetate, camsylate, carbonate, chloride, clavulanate, citrate, dihydrochloride, edetate, edisylate, estolate, esylate, fumarate, gluceptate, gluconate, glutamate, glycollylarsanilate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, hydroxynapthoate, iodide, isothionate, lactate, lactobionate, laurate, malate, maleate, mandelate, mesylate, methylsulfate, mucate, napsylate, nitrate, oleate, oxalate, pamaote, palmitate, panthothenate, phosphate/diphosphate, polygalacturonate, salicylate, stearate, subacetate, succinate, tannate, tartrate, teoclate, tosylate, triethiodide, and valerate.

Ester derivatives of the described HMG-CoA reductase inhibitor compounds may act as prodrugs which, when absorbed into the bloodstream of a warm-blooded animal, may cleave in such a manner as to release the drug form and permit the drug to afford improved therapeutic efficacy.

"Prenyl-protein transferase inhibitor" refers to a compound which inhibits any one or any combination of the prenyl-protein transferase enzymes, including farnesyl-protein transferase (FPTase), geranylgeranyl-protein transferase type I (GGPTase-I), and geranylgeranyl-protein transferase type-II (GGPTase-II, also called Rab GGPTase). Examples of prenyl-protein transferase inhibiting compounds include (.+-.)-6-[amino(4-chlorophenyl)(1-methyl-1H-imidazol-5-yl)methyl]-4-(3-chl orophenyl)-1-methyl-2(1H)-quinolinone, (-)-6-[amino(4-chlorophenyl)(1-methyl-1H-imidazol-5-yl)methyl]-4-(3-chloro phenyl)-1-methyl-2(1H)-quinolinone, (+)-6-[amino(4-chlorophenyl)(1-methyl-1H-imidazol-5-yl) methyl]-4-(3-chlorophenyl)-1-methyl-2(1H)-quinolinone, 5(S)-n-butyl-1-(2,3-dimethylphenyl)-4-[1-(4-cyanobenzyl)-5-imidazolylmethy l]-2-piperazinone, (S)-1-(3-chlorophenyl)-4-[1-(4-cyanobenzyl)-5-imidazolylmethyl]-5-[2-(etha nesulfonyl) methyl)-2-piperazinone, 5(S)-n-Butyl-1-(2-methylphenyl)-4-[1-(4-cyanobenzyl)-5-imidazolylmethyl]-2 -piperazinone, 1-(3-chlorophenyl)-4-[1-(4-cyanobenzyl)-2-methyl-5-imidazolylmethyl]-2-pip erazinone, 1-(2,2-diphenylethyl)-3-[N-(1-(4-cyanobenzyl)-1H-imidazol-5-ylethyl)carbam oyl]piperidine, 4-{5-[4-hydroxymethyl-4-(4-chloropyridin-2-ylmethyl)-piperidine-1-ylmethyl ]-2-methylimidazol-1-ylmethyl}benzonitrile, 4-{5-[4-hydroxymethyl-4-(3-chlorobenzyl)-piperidine-1-ylmethyl]-2-methylim idazol-1-ylmethyl}benzonitrile, 4-{3-[4-(2-oxo-2H-pyridin-1-yl)benzyl]-3H-imidazol-4-ylmethyl}benzonitrile , 4-{3-[4-(5-chloro-2-oxo-2H-[1,2']bipyridin-5'-ylmethyl]-3H-imidazol-ylmeth yl}benzonitrile, 4-{3-[4-(2-oxo-2H-[1,2']bipyridin-5'-ylmethyl]-3H-imidazol-4-ylmethyl}benz onitrile, 4-[3-(2-oxo-1-phenyl-1,2-dihydropyridin-4-ylmethyl)-3H-imidazol-4-ylmethyl }benzonitrile, 18,19-dihydro-19-oxo-5H,17H-6,10: 12,16-dimetheno-1H-imidazo[4,3-c][1,11,4]dioxaazacyclo-nonadecine-9-carbon itrile, (.+-.)-19,20-dihydro-19-oxo-5H-18,21-ethano-12,14-etheno-6,10-metheno-22H- benzo[d]imidazo[4,3-k][1,6,9,12]oxatriaza-cyclooctadecine-9-carbonitrile, 19,20-dihydro-19-oxo-5H,17H-18,21-ethano-6,10: 12,16-dimetheno-22H-imidazo[3,4-h][1,8,11,14]oxatriazacycloeicosine-9-carb onitrile, and (.+-.)-19,20-dihydro-3-methyl-19-oxo-5H-18,21-ethano-12,14-etheno-6,10-met heno-22H-benzo[d]imidazo[4,3-k][1,6,9,12]oxa-triazacyclooctadecine-9-carbon itrile.

Other examples of prenyl-protein transferase inhibitors can be found in the following publications and patents: WO 96/30343, WO 97/18813, WO 97/21701, WO 97/23478, WO 97/38665, WO 98/28980, WO 98/29119, WO 95/32987, U.S. Pat. No. 5,420,245, U.S. Pat. No. 5,523,430, U.S. Pat. No. 5,532,359, U.S. Pat. No. 5,510,510, U.S. Pat. No. 5,589,485, U.S. Pat. No. 5,602,098, European Patent Publ. 0 618 221, European Patent Publ. 0 675 112, European Patent Publ. 0 604 181, European Patent Publ. 0 696 593, WO 94/19357, WO 95/08542, WO 95/11917, WO 95/12612, WO 95/12572, WO 95/10514, U.S. Pat. No. 5,661,152, WO 95/10515, WO 95/10516, WO 95/24612, WO 95/34535, WO 95/25086, WO 96/05529, WO 96/06138, WO 96/06193, WO 96/16443, WO 96/21701, WO 96/21456, WO 96/22278, WO 96/24611, WO 96/24612, WO 96/05168, WO 96/05169, WO 96/00736, U.S. Pat. No. 5,571,792, WO 96/17861, WO 96/33159, WO 96/34850, WO 96/34851, WO 96/30017, WO 96/30018, WO 96/30362, WO 96/30363, WO 96/31111, WO 96/31477, WO 96/31478, WO 96/31501, WO 97/00252, WO 97/03047, WO 97/03050, WO 97/04785, WO 97/02920, WO 97/17070, WO 97/23478, WO 97/26246, WO 97/30053, WO 97/44350, WO 98/02436, and U.S. Pat. No. 5,532,359. For an example of the role of a prenyl-protein transferase inhibitor on angiogenesis see European J. of Cancer, Vol. 35, No. 9, pp. 1394-1401 (1999).

Examples of HIV protease inhibitors include amprenavir, abacavir, CGP-73547, CGP-61755, DMP-450, indinavir, nelfinavir, tipranavir, ritonavir, saquinavir, ABT-378, AG 1776, and BMS-232,632. Examples of reverse transcriptase inhibitors include delaviridine, efavirenz, GS-840, HB Y097, lamivudine, nevirapine, AZT, 3TC, ddC, and ddI.

"Angiogenesis inhibitors" refers to compounds that inhibit the formation of new blood vessels, regardless of mechanism. Examples of angiogenesis inhibitors include, but are not limited to, tyrosine kinase inhibitors, such as inhibitors of the tyrosine kinase receptors Flt-1 (VEGFR1) and Flk-1/KDR (VEGFR20), inhibitors of epidermal-derived, fibroblast-derived, or platelet derived growth factors, MMP (matrix metalloprotease) inhibitors, integrin blockers, interferon-.alpha., interleukin-12, pentosan polysulfate, cyclooxygenase inhibitors, including nonsteroidal anti-inflammatories (NSAIDs) like aspirin and ibuprofen as well as selective cyclooxygenase-2 inhibitors like celecoxib and rofecoxib (PNAS, Vol. 89, p. 7384 (1992); JNCI, Vol. 69, p. 475 (1982); Arch. Opthalmol., Vol. 108, p. 573 (1990); Anat. Rec., Vol. 238, p. 68 (1994); FEBS Letters, Vol. 372, p. 83 (1995); Clin, Orthop. Vol. 313, p. 76 (1995); J. Mol. Endocrinol., Vol. 16, p. 107 (1996); Jpn. J. Pharmacol., Vol. 75, p. 105 (1997); Cancer Res., Vol. 57, p. 1625 (1997); Cell, Vol. 93, p. 705 (1998); Intl. J. Mol. Med., Vol. 2, p. 715 (1998); J. Biol. Chem., Vol. 274, p. 9116 (1999)), carboxyamidotriazole, combretastatin A-4, squalamine, 6-O-chloroacetyl-carbonyl)-fumagillol, thalidomide, angiostatin, troponin-1, angiotensin II antagonists (see Fernandez et al., J. Lab. Clin. Med. 105:141-145 (1985)), and antibodies to VEGF (see, Nature Biotechnology, Vol. 17, pp. 963-968 (October 1999); Kim et al., Nature, 362, 841-844 (1993); WO 00/44777; and WO 00/61186).

As described above, the combinations with NSAID's are directed to the use of NSAID's which are potent COX-2 inhibiting agents. For purposes of this specification an NSAID is potent if it possess an IC.sub.50 for the inhibition of COX-2 of 1 .mu.M or less as measured by cell or microsomal assays.

The invention also encompasses combinations with NSAID's which are selective COX-2 inhibitors. For purposes of this specification NSAID's which are selective inhibitors of COX-2 are defined as those which possess a specificity for inhibiting COX-2 over COX-1 of at least 100 fold as measured by the ratio of IC.sub.50 for COX-2 over IC.sub.50 for COX-1 evaluated by cell or microsomal assays. Such compounds include, but are not limited to those disclosed in U.S. Pat. No. 5,474,995, issued Dec. 12, 1995, U.S. Pat. No. 5,861,419, issued Jan. 19, 1999, U.S. Pat. No. 6,001,843, issued Dec. 14, 1999, U.S. Pat. No. 6,020,343, issued Feb. 1, 2000, U.S. Pat. No. 5,409,944, issued Apr. 25, 1995, U.S. Pat. No. 5,436,265, issued Jul. 25, 1995, U.S. Pat. No. 5,536,752, issued Jul. 16, 1996, U.S. Pat. No. 5,550,142, issued Aug. 27, 1996, U.S. Pat. No. 5,604,260, issued Feb. 18, 1997, U.S. Pat. No. 5,698,584, issued Dec. 16, 1997, U.S. Pat. No. 5,710,140, issued Jan. 20, 1998, WO 94/15932, published Jul. 21, 1994, U.S. Pat. No. 5,344,991, issued Jun. 6, 1994, U.S. Pat. No. 5,134,142, issued Jul. 28, 1992, U.S. Pat. No. 5,380,738, issued Jan. 10, 1995, U.S. Pat. No. 5,393,790, issued Feb. 20, 1995, U.S. Pat. No. 5,466,823, issued Nov. 14, 1995, U.S. Pat. No. 5,633,272, issued May 27, 1997, and U.S. Pat. No. 5,932,598, issued Aug. 3, 1999, all of which are hereby incorporated by reference.

Other examples of specific inhibitors of COX-2 include the following: 3-(3-fluorophenyl)-4-(4-(methylsulfonyl)phenyl)-2-(5H)-furanone; 3-(3,4-difluorophenyl)-4-(4-(methylsulfonyl)phenyl)-2-(5H)-furanone; 3-(3,4-dichlorophenyl)-4-(4-(methylsulfonyl)phenyl)-2-(5H)-furanone; 3-phenyl-4-(4-(methylsulfonyl)phenyl)-2-(5H)-furanone; 5,5-dimethyl-3-(3-fluorophenyl)-4-(methylsulfonyl)phenyl)-2-(5H)-furanone; 3-(4-methylsulfonyl)phenyl-2-phenyl-5-trifluoromethylpyridine; 2-(3-chlorophenyl)-3-(4-methylsulfonyl)phenyl-5-trifluoromethyl-pyridine; 2-(4-chlorophenyl)-3-(4-methylsulfonyl)phenyl-5-trifluoromethyl-pyridine; 2-(4-fluorophenyl)-3-(4-methylsulfonyl)phenyl-5-trifluoromethyl-pyridine; 3-(4-methylsulfonyl)phenyl-2-(3-pyridinyl)-5-trifluoromethylpyridine; 5-methyl-3-(4-methylsulfonyl)phenyl-2-phenylpyridine; 2-(4-chlorophenyl)-5-methyl-3-(4-methylsulfonyl)phenylpyridine; 5-methyl-3-(4-methylsulfonyl)phenyl-2-(3-pyridinyl)pyridine; 5-chloro-2-(4-chlorophenyl)-3-(4-methylsulfonyl)phenylpyridine; 5-chloro-3-(4-methylsulfonyl)phenyl-2-(2-pyridinyl)pyridine; 5-chloro-3-(4-methylsulfonyl)phenyl-2-(3-pyridinyl)pyridine; 5-chloro-3-(4-methylsulfonyl)phenyl-2-(4-pyridinyl)pyridine; 5-chloro-3-(4-methylsulfonyl)phenyl-2-(2-methyl-5-pyridinyl)pyridine; 2-(4-chlorophenyl)-3-(4-methylsulfonyl)phenylpyridinyl-5-carboxylic acid methyl ester; 2-(4-chlorophenyl)-3-(4-methylsulfonyl)phenylpyridinyl-5-carboxylic acid; 5-cyano-2-(4-chlorophenyl)-3-(4-methylsulfonyl)phenylpyridine; 5-chloro-3-(4-methylsulfonyl)phenyl-2-(3-pyridyl)pyridine hydromethanesulfonate; 5-chloro-3-(4-methylsulfonyl)phenyl-2-(3-pyridyl)pyridine hydrochloride; 5-chloro-3-(4-methylsulfonyl)phenyl-2-(2-methyl-5-pyridinyl)pyridine hydrochloride; 5-chloro-3-(4-methylsulfonyl)phenyl-2-(2-ethyl-5-pyridinyl)pyridine; 5-chloro-3-(4-methylsulfonyl)phenyl-2-(2-ethyl-5-pyridinyl)pyridine hydromethanesulfonate; 3-(3,4-difluorophenoxy)-5,5-dimethyl-4-(4-(methylsulfonyl)phenyl)-5H-furan- 2-one; 3-(3-fluorophenoxy)-5,5-dimethyl-4-(4-(methylsulfonyl)phenyl)-5H-furan-2-on e; 3-(3,5-difluorophenoxy)-5,5-dimethyl-4-(methylsulfonyl)phenyl)-5H-furan-2-o ne; 3-phenoxy-5,5-dimethyl-4-(4-(methylsulfonyl)phenyl)-5H-furan-2-one; 3-(2,4-difluorophenoxy)-5,5-dimethyl-4-(4-(methylsulfonyl)phenyl)-5H-furan- 2-one; 3-(4-chlorophenoxy)-5,5-dimethyl-4-(4-(methylsulfonyl)phenyl)-5H-furan-2-on e; 3-(3,4-dichlorophenoxy)-5,5-dimethyl-4-(methylsulfonyl)phenyl)-5H-furan-2-o ne; 3-(4-fluorophenoxy)-5,5-dimethyl-4-(4-(methylsulfonyl)phenyl)-5H-furan-2-on e; 3-(4-fluorophenylthio)-5,5-dimethyl-4-(4-(methylsulfonyl)phenyl)-5H-furan-2 -one; 3-(3,5-difluorophenylthio)-5,5-dimethyl-4-(4-(methylsulfonyl)phenyl)-5H-fur an-2-one; 3-phenylthio-5,5-dimethyl-4-(4-(methylsulfonyl)phenyl)-5H-furan-2-one; 3-(N-phenylamino)-5,5-dimethyl-4-(4-(methylsulfonyl)phenyl)-5H-furan-2-one; 3-N-methyl-N-phenylamino)-5,5-dimethyl-4-(4-(methylsulfonyl)phenyl)-5H-fura n-2-one; 3-cyclohexyloxy-5,5-dimethyl-4-(4-(methylsulfonyl)phenyl)-5H-furan-2-one; 3-phenylthio-4-(4-(methylsulfonyl)phenyl)-5H-furan-2-one; 3-benzyl-5,5-dimethyl-4-(4-(methylsulfonyl)phenyl)-5H-furan-2-one; 3-(3,4-difluorophenylhydroxymethyl)-5,5-dimethyl-4-(4-(methylsulfonyl)pheny l)-5H-furan-2-one; 3-(3,4-difluorobenzoyl)-5,5-dimethyl-4-(4-(methylsulfonyl)phenyl)-5H-furan- 2-one; 3-benzoyl-5,5-dimethyl-4-(4-(methylsulfonyl)phenyl)-5H-furan-2-one; 4-(4-(methylsulfonyl)phenyl)-3-phenoxy-1-oxaspiro[4.4]non-3-en-2-one; 4-(4-(methylsulfonyl)phenyl)-3-phenylthio-1-oxaspiro[4.4]non-3-en-2-one; 4-(2-oxo-3-phenylthio-1-oxa-spiro[4,4]non-3-en-4-yl)benzenesulfonamide; 3-(4-fluorobenzyl)-5,5-dimethyl-4-(4-(methylsulfonyl)phenyl)-5H-furan-2-one ; 3-(3,4-difluorophenoxy)-5-methoxy-5-methyl-4-(4-(methylsulfonyl)phenyl)-5H- furan-2-one; 3-(5-chloro-2-pyridyloxy)-5,5-dimethyl-4-(4-(methylsulfonyl)phenyl)-5H-fura n-2-one; 3-(2-pyridyloxy)-5,5-dimethyl-4-(4-(methylsulfonyl)phenyl)-5H-furan-2-one; 3-(6-methyl-2-pyridyloxy)-5,5-dimethyl-4-(4-(methylsulfonyl)phenyl)-5H-fura n-2-one; 3-(3-isoquinolinoxy)-5,5-dimethyl-4-(4-(methylsulfonyl)phenyl)-5H-furan-2-o ne; 3-(4-(methylsulfonyl)phenyl)-2-phenoxycyclopent-2-enone; 3-(4-(methylsulfonyl)phenyl)-2-(3,4-difluorophenoxy)cyclopent-2-enone; 5,5-dimethyl-4-(4-methylsulfonylphenyl)-3-(5-bromopyridin-2-yloxy)-5H-furan -2-one; 5,5-dimethyl-4-(4-methylsulfonylphenyl)-3-(2-propoxy)-5H-furan-2-one; 2-(3,4-difluorophenoxy)-3-(4-methylsulfonylphenyl)-cyclopent-2-enone; 3-(5-benzothiophenyloxy)-5,5-dimethyl-4-(4-(methylsulfonyl)phenyl)-5H-furan -2-one; 5,5-dimethyl-4-(4-methylsulfonyl-phenyl)-3-(pyridyl-4-oxy)-5H-furan-2-one; 5,5-methyl-4-(4-methylsulfonyl-phenyl)-3-(pyridyl-3-oxy)-5H-furan-2-one; 3-(2-methyl-5-pyridyloxy)-5,5-dimethyl-4-(4-(methylsulfonyl)phenyl)-5H-fura n-2-one; 3-(2-fluoro-4-fluoromethyl)phenoxy-4-(4-methylsulfonyl)phenyl)-5,5-dimethyl -5H-furan-2-one; 3-(5-chloro-2-pyridylthio)-5,5-dimethyl-4-(4-methylsulfonyl)phenyl-5H-furan -2-one; 2-(3,5-difluorophenoxy)-3-(4-methylsulfonylphenyl)-cyclopent-2-enone; 3-(2-pyrimidinoxy)-5,5-dimethyl-4-(4-methylsulfonyl)phenyl-5H-furan-2-one; 3-(3-methyl-2-pyridyloxy)-5,5-dimethyl-4-(4-methylsulfonyl)phenyl-5H-furan- 2-one; 3-(3-chloro-5-pyridyloxy)-5,5-dimethyl-4-(4-(methylsulfonyl)phenyl)-5H-fura n-2-one; 3-(3-(1,2,5-thiadiazolyl)oxy)-4-(4-(methylsulfonyl)phenyl)-5,5-dimethyl-5H- furan-2-one; 3-(5-isoquinohnoxy)-5,5-dimethyl-4-(4-methylsulfonyl)phenyl-5H-furan-2-one; 3-(6-amino-2-pyridyloxy)-5,5-dimethyl-4-(4-(methylsulfonyl)phenyl)-5H-furan -2-one; 3-(3-chloro-4-fluoro)phenoxy-4-(methylsulfonyl)phenyl)-5,5-dimethyl-5H-fura n-2-one; 3-(6-quinolinoxy)-5,5-dimethyl-4-(4-(methylsulfonyl)phenyl)-5H-furan-2-one; 3-(5-nitro-2-pyridyloxy)-5,5-dimethyl-4-(4-methylsulfonyl)phenyl-5H-furan-2 -one; 3-(2-thiazolylthio)-5,5-dimethyl-4-(4-(methylsulfonyl)phenyl)-5H-furan-2-on e; 3-(3-chloro-5-pyridyloxy)-5,5-dimethyl-4-(4-(methylsulfonyl)phenyl)-5H-fura n-2-one; 5,5-dimethyl-4-(4-methylsulfonylphenyl)-3-(2-propoxy)-5H-furan-2-one; 3-(3-trifluoromethyl)phenoxy-4-(4-methylsulfonyl)phenyl)-5,5-dimethyl-5H-fu ran-2-one; 5,5-dimethyl-(4-(4-methylsulfonyl)phenyl)-3-(piperidine-1-carbonyl)-5-H-fur an-2-one; 5,5-dimethyl-3-(2-Butoxy)-4-(4-methylsulfonylphenyl)-5H-furan-2-one; 5,5-dimethyl-4-(4-methylsulfonylphenyl)-3-(3-pentoxy)-5H-furan-2-one; 2-(5-chloro-2-pyridyloxy)-3-(4-methylsulfonyl)phenylcyclopent-2-enone; 3-(4-methyl-2-pyridyloxy)-5,5-dimethyl-4-(4-methylsulfonyl)phenyl-5H-furan- 2-one; (5R)-3-(3,4-difluorophenoxy)-5-ethyl-5-methyl-4-(4-methylsulfonyl)phenyl-5H -furan-2-one; (5R)-3-(4-chlorophenoxy)-5-ethyl-5-methyl-4-(4-methylsulfonyl)phenyl-5H-fur an-2-one; 3-(2-methyl-3-pyridyloxy)-5,5-dimethyl-4-(4-methylsulfonyl)phenyl-5H-furan- 2-one; 3-(4-methyl-5-nitro-2-pyridyloxy)-5,5-dimethyl-4-(4-methylsulfonyl)phenyl-5 H-furan-2-one; 3-(5-chloro-4-methyl-2-pyridyloxy)-5,5-dimethyl-4-(4-methylsulfonyl)phenyl- 5H-furan-2-one; 3-(5-fluoro-4-methyl-2-pyridyloxy)-5,5-dimethyl-4-(4-methylsulfonyl)phenyl- 5H-furan-2-one; 3-(3-chloro-2-pyridyloxy)-5,5-dimethyl-4-(4-methylsulfonyl)phenyl-5H-furan- 2-one; 3-(4-fluorophenoxy)-5-methyl-4-(4-methylsulfonyl)phenyl-5-propyl-5H-furan-2 -one; 3-(N,N-diethylamino)-5,5-dimethyl-4-(4-(methylsulfonyl)phenyl)-5H-furan-2-o ne; 5,5-dimethyl-4-(4-methylsulfonyl-phenyl)-3-(3,5-dichloro-2-pyridyloxy)-5H-f uran-2-one; (5R)-3-(4-bromophenoxy)-5-ethyl-5-methyl-4-(4-methylsulfonyl)phenyl-5H-fura n-2-one; (5R)-3-(4-methoxyphenoxy)-5-ethyl-5-methyl-4-(4-methylsulfonyl)phenyl-5H-fu ran-2-one; (5R)-3-(5-chloro-2-pyridyloxy)-5-methyl-4-(4-methylsulfonyl)phenyl-5-(2,2,2 -trifluoroethyl)-5H-furan-2-one; 3-(5-chloro-2-pyridyloxy)-5-methyl-4-(4-methylsulfonyl)phenyl-5-propyl-5H-f uran-2-one; 3-(1-cyclopropyl-ethoxy)-5,5-dimethyl-4-(4-methylsulfonyl)phenyl)-5H-furan- 2-one; 5-methyl-4-(4-(methylsulfonyl)phenyl)-3-(2-(propoxy)-5-(2-trifluoroethyl)-5 H-furan-2-one; 5(R)-5-ethyl-5-methyl-4-(4-(methylsulfonyl)phenyl)-3-(2-propoxy)-5H-furan-2 -one; 5,5-dimethyl-3-(2,2-dimethylpropyloxy)-4-(4-(methylsulfonyl)phenyl)-5H-fura n-2-one; 5R)-3-(1-cyclopropyl-ethoxy)-5-ethyl-5-methyl-4-(4-(methylsulfonyl)phenyl-5 H-furan-2-one; 5(S)-5-ethyl-5-methyl-4-(4-(methylsulfonyl)phenyl-3-(2-propoxy)-5H-furan-2- one; 3-(1-cyclopropylethoxy)-5,5-dimethyl-4-(4-(methylsulfonyl)phenyl)-5H-furan- 2-one; 3-(1-cyclopropylethoxy)-5,5-dimethyl-4-(4-(methylsulfonyl)phenyl)-5H-furan- 2-one; 5,5-dimethyl-3-(isobutoxy)-4-(4-(methylsulfonyl)phenyl)-5H-furan-2-one; 3-(4-bromophenoxy)-5,5-dimethyl-4-(4-(methylsulfonyl)phenyl)-5H-furan-2-one ; 3-(2-quinolinoxy)-5,5-dimethyl-4-(4-(methylsulfonyl)phenyl)-5H-furan-2-one; 3-(2-chloro-5-pyridyloxy)-5,5-dimethyl-4-(4-(methylsulfonyl)phenyl)-5H-fura n-2-one; 3-(6-benzothiazolyloxy)-5,5-dimethyl-4-(4-(methylsulfonyl)phenyl)-5H-furan- 2-one; 3-(6-chloro-2-pyridyloxy)-5,5-dimethyl-4-(4-(methylsulfonyl)phenyl)-5H-fura n-2-one; 3-(4-quinazolyloxy)-5,5-dimethyl-4-(4-(methylsulfonyl)phenyl)-5H-furan-2-on e; (5R)-3-(5-fluoro-2-pyridyloxy)-5-ethyl-5-methyl-4-(4-methylsulfonyl)phenyl- 5H-furan-2-one; (5R)-3-(4-fluorophenoxy)-5-ethyl-5-methyl-4-(4-methylsulfonyl)phenyl-5H-fur an-2-one; (5R)-3-(5-fluoro-2-pyridyloxy)-5-methyl-4-(4-methylsulfonyl)phenyl-5-(2,2,2 -trifluoroethyl)-5H-furan-2-one; 3-(1-isoquinolinyloxy)-5,5-dimethyl-4-(methylsulfonyl)phenyl-5H-furan-2-one ; (5R)-3-(4-fluorophenoxy)-5-methyl-4-(4-methylsulfonyl)phenyl-5-(2,2,2-trifl uoroethyl)-5H-furan-2-one; 3-(3-fluoro-2-pyridyloxy)-5,5-dimethyl-4-(4-methylsulfonyl)phenyl-5H-furan- 2-one; (5R)-3-(3,4-difluorophenoxy)-5-methyl-4-(4-methylsulfonyl)phenyl-5-(2,2,2-t rifluoroethyl)-5H-furan-2-one; (5R)-3-(5-chloro-2-pyridyloxy)-5-ethyl-5-methyl-4-(4-methylsulfonyl)phenyl- 5H-furan-2-one; 3-(3,4-difluorophenoxy)-5-methyl-5-trifluoromethyl-4-(4-methylsulfonyl)phen yl-5H-furan-2-one; 3-(3,4-fluorophenoxy)-5-methyl-4-(4-(methylsulfonyl)phenyl)-5-propyl-5H-fur an-2-one; 3-cyclobutyloxy-5,5-dimethyl-4-(4-methylsulfonylphenyl-5H-furan-2-one; 3-(1-indanyloxy)-5,5-dimethyl-4-(4-(methylsulfonyl)phenyl)-5H-furan-2-one; 3-(2-indanyloxy)-5,5-dimethyl-4-(4-methylsulfonyl)phenyl)-5H-furan-2-one; 3-cyclopentyloxy-5,5-dimethyl-4-(4-methylsulfonylphenyl)5H-furan-2-one; 3-(3,3-dimethylcyclopentyloxy)-5,5-dimethyl-4-(4-methylsulfonyl-phenyl)-5H- furan-2-one; 3-isopropoxy-5-methyl-4-(4-methylsulfonylphenyl)-5-propyl-5H-furan-2-one; 3-(2-methoxy-5-pyridyloxy)-5,5-dimethyl-4-(4-methylsulfonyl)phenyl-5H-furan -2-one; 3-(5-methyl-2-pyridyloxy)-5,5-dimethyl-4-(4-methylsulfonyl)phenyl-5H-furan- 2-one; (5RS)-3-(3,4-difluorophenoxy)-5-methyl-4-(4-methylsulfonyl)phenyl-5-(2,2,2- trifluoroethyl)-5H-furan-2-one; 3-(3-chloro-4-methoxyphenoxy)-5,5-dimethyl-4-(4-methylsulfonyl)phenyl-5H-fu ran-2-one; (5R)-3-(3-chloro-4-methoxyphenoxy)-5-ethyl-5-methyl-4-(4-methylsulfonyl)phe nyl-5H-furan-2-one; (5R)-3-(4-chlorophenoxy)-5-trifluoroethyl-5-methyl-4-(4-methylsulfonyl)phen yl-5H-furan-2-one; (5R)-3-(4-bromophenoxy)-5-trifluoroethyl-5-methyl-4-(4-methylsulfonyl)pheny l-5H-furan-2-one; 5-cyclopropylmethyl-3-(3,4-difluorophenoxy)-5-methyl-(4-methylsulfonyl)phen yl-5H-furan-2-one; (5R)-3-(3-fluorophenoxy)-5-ethyl-5-methyl-4-(4-methylsulfonyl)phenyl-5H-fur an-2-one; (5R)-3-(4-chloro-3-fluorophenoxy)-5-ethyl-5-methyl-4-(4-methylsulfonyl)phen yl-5H-furan-2-one; (5R)-3-phenoxy-5-ethyl-5-methyl-4-(4-methylsulfonyl)phenyl-5H-furan-2-one; (5R)-3-(4-chloro-3-methylphenoxy)-5-ethyl-5-methyl-4-(4-methylsulfonyl)phen yl-5H-furan-2-one; 3-(4-chloro-3-methylphenoxy)-5-5-dimethyl-4-(4-methylsulfonyl)phenyl-5H-fur an-2-one; (5R)-3-(5-bromo-2-pyridyloxy)-4-(4-methylsulfonylphenyl)-5-methyl-5-(2,2,2- trifluoroethyl)-5H-furan-2-one; (5R)-3-(5-bromo-2-pyridyloxy)-4-(4-methylsulfonylphenyl)-5-ethyl-5-methyl-5 H-furan-2-one; 3-(5-chloro-6-methyl-2-pyridyloxy)-5,5-dimethyl-4-(4-methylsulfonyl)phenyl- 5H-furan-2-one; 3-(5-cyclopropyl-2-pyridyloxy)-5,5-dimethyl-4-(4-methylsulfonyl)phenyl-5H-f uran-2-one; 3-(1-cyclopropylethoxy)-4-(4-methylsulfonyl)phenyl-5H-furan-2-one; and 3-(cyclopropylmethoxy)-4-(4-methylsulfonyl)phenyl-5H-furan-2-one;

or a pharmaceutically acceptable salt or stereoisomer thereof.

Inhibitors of COX-2 that are particularly useful in the instant method of treatment are: 3-phenyl-4-(4-(methylsulfonyl)phenyl)-2-(5H)-furanone; and ##STR4## 5-chloro-3-(4-methylsulfonyl)phenyl-2-(2-methyl-5-pyridinyl)pyridine; ##STR5##

or a pharmaceutically acceptable salt thereof.

General and specific synthetic procedures for the preparation of the COX-2 inhibitor compounds described above are found in U.S. Pat. No. 5,474,995, issued Dec. 12, 1995, U.S. Pat. No. 5,861,419, issued Jan. 19, 1999, and U.S. Pat. No. 6,001,843, issued Dec. 14, 1999, all of which are herein incorporated by reference.

Compounds that have been described as specific inhibitors of COX-2 and are therefore useful in the present invention include, but are not limited to, the following: ##STR6##

or a pharmaceutically acceptable salt thereof.

Compounds which are described as specific inhibitors of COX-2 and are therefore useful in the present invention, and methods of synthesis thereof, can be found in the following patents, pending applications and publications, which are herein incorporated by reference: WO 94/15932, published Jul. 21, 1994, U.S. Pat. No. 5,344,991, issued Jun. 6, 1994, U.S. Pat. No. 5,134,142, issued Jul. 28, 1992, U.S. Pat. No. 5,380,738, issued Jan. 10, 1995, U.S. Pat. No. 5,393,790, issued Feb. 20, 1995, U.S. Pat. No. 5,466,823, issued Nov. 14, 1995, U.S. Pat. No. 5,633,272, issued May 27, 1997, and U.S. Pat. No. 5,932,598, issued Aug. 3, 1999.

Compounds which are specific inhibitors of COX-2 and are therefore useful in the present invention, and methods of synthesis thereof, can be found in the following patents, pending applications and publications, which are herein incorporated by reference: U.S. Pat. No. 5,474,995, issued Dec. 12, 1995, U.S. Pat. No. 5,861,419, issued Jan. 19, 1999, U.S. Pat. No. 6,001,843, issued Dec. 14, 1999, U.S. Pat. No. 6,020,343, issued Feb. 1, 2000, U.S. Pat. No. 5,409,944, issued Apr. 25, 1995, U.S. Pat. No. 5,436,265, issued Jul. 25, 1995, U.S. Pat. No. 5,536,752, issued Jul. 16, 1996, U.S. Pat. No. 5,550,142, issued Aug. 27, 1996, U.S. Pat. No. 5,604,260, issued Feb. 18, 1997, U.S. Pat. No. 5,698,584, issued Dec. 16, 1997, and U.S. Pat. No. 5,710,140, issued Jan. 20, 1998.

Other examples of angiogenesis inhibitors include, but are not limited to, endostation, ukrain, ranpirnase, IM862, 5-methoxy-4-[2-methyl-3-(3-methyl-2-butenyl)oxiranyl]-1-oxaspiro[2,5]oct-6 -yl(chloroacetyl)carbamate, acetyldinanaline, 5-amino-1-[[3,5-dichloro-4-(4-chlorobenzoyl)phenyl]methyl]-1H-1,2,3-triazo le-4-carboxamide, CM101, squalamine, combretastatin, RPI4610, NX31838, sulfated mannopentaose phosphate, 7,7-(carbonyl-bis[imino-N-methyl-4,2-pyrrolocarbonyl-imino[N-methyl-4,2-py rrole]-carbonylimino]-bis-(1,3-naphthalene disulfonate), and 3-[(2,4-dimethylpyrrol-5-yl)methylene]-2-indolinone (SU5416).

As used above, "integrin blockers" refers to compounds which selectively antagonize, inhibit or counteract binding of a physiological ligand to the .alpha..sub.v.beta..sub.3 integrin, to compounds which selectively antagonize, inhibit or counteract binding of a physiological ligand to the .alpha.v.beta.5 integrin, to compounds which antagonize, inhibit or counteract binding of a physiological ligand to both the .alpha..sub.v.beta..sub.3 integrin and the .alpha..sub.v.beta..sub.5 integrin, and to compounds which antagonize, inhibit or counteract the activity of the particular integrin(s) expressed on capillary endothelial cells. The term also refers to antagonists of the .alpha..sub.v.beta..sub.6, .alpha..sub.v.beta..sub.8, .alpha..sub.1.beta..sub.1, .alpha..sub.2.beta..sub.1, .alpha..sub.5.beta..sub.1, .alpha..sub.6.beta..sub.1 and .alpha..sub.6.beta..sub.4 integrins. The term also refers to antagonists of any combination of .alpha..sub.v.beta..sub.3, .alpha..sub.v.beta..sub.5, .alpha..sub.v.beta..sub.6, .alpha..sub.v.mu..sub.8, .alpha..sub.1.beta..sub.1, .alpha..sub.2.beta..sub.1, .alpha..sub.5.beta..sub.1, .alpha..sub.6.beta..sub.1 and .alpha..sub.6.beta..sub.4 integrins.

Some specific examples of tyrosine kinase inhibitors include N-(trifluoromethylphenyl)-5-methylisoxazol-4-carboxamide, 3-[(2,4-dimethylpyrrol-5-yl)methylidenyl)indolin-2-one, 17-(allylamino)-17-demethoxygeldanamycin, 4-(3-chloro-4-fluorophenylamino)-7-methoxy-6-[3-(4-morpholinyl)propoxyl]qu inazoline, N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine, BIBX1382, 2,3,9,10,11,12-hexahydro-10-(hydroxymethyl)-10-hydroxy-9-methyl-9,12-epoxy -1H-diindolo[1,2,3-fg:3',2',1'-kl]pyrrolo[3,4-i][1,6]benzodiazocin-1-one, SH268, genistein, STI571, CEP2563, 4-(3-chlorophenylamino)-5,6-dimethyl-7H-pyrrolo[2,3-d]pyrimidinemethane sulfonate, 4-(3-bromo-4-hydroxyphenyl)amino-6,7-dimethoxyquinazoline, 4-(4'-hydroxyphenyl)amino-6,7-dimethoxyquinazoline, SU6668, STI571A, N-4-chlorophenyl-4-(4-pyridylmethyl)-1-phthalazinamine, and EMD121974.

The instant compounds are also useful, alone or in combination with platelet fibrinogen receptor (GP IIb/IIIa) antagonists, such as tirofiban, to inhibit metastasis of cancerous cells. Tumor cells can activate platelets largely via thrombin generation. This activation is associated with the release of VEGF. The release of VEGF enhances metastasis by increasing extravasation at points of adhesion to vascular endothelium (Amirkhosravi, Platelets 10, 285-292, 1999). Therefore, the present compounds can serve to inhibit metastasis, alone or in combination with GP IIb/IIIa) antagonists. Examples of other fibrinogen receptor antagonists include abciximab, eptifibatide, sibrafiban, lamifiban, lotrafiban, cromofiban, and CT50352.

Combinations with compounds other than anti-cancer compounds are also encompassed to treat conditions other than cancer. For example, combinations of the instantly claimed compounds with PPAR-.gamma. (i.e., PPAR-gamma) agonists are useful in the treatment of diabetic retinopathy. PPAR-.gamma. is the nuclear peroxisome proliferator-activated receptor .gamma.. The expression of PPAR-.gamma. on endothelial cells and its involvement in angiogenesis in corneal and choroidal experimental systems has been reported in the literature (see J. Cardiovasc. Pharmacol. 1998; 31:909-913; J. Biol. Chem. 1999; 274:9116-9121; Invest. Ophthalmol Vis. Sci. 2000; 41:2309-2317). More recently, PPAR-.gamma. agonists have been shown to inhibit the angiogenic response to VEGF in vitro; both troglitazone and rosiglitazone maleate inhibit the development of retinal neovascularization in mice. (Arch. Ophthamol. 2001; 119:709-717). Examples of PPAR-.gamma. aganoists and PPAR-.gamma./.alpha. aganoists include, but are not limited to, thiazolidinediones (such as DRF2725, CS-011, troglitazone, rosiglitazone, and pioglitazone), fenofibrate, gemfibrozil, clofibrate, GW2570, SB219994, AR-H039242, JTT-501, MCC-555, GW2331, GW409544, NN2344, KRP297, NP0110, DRF4158, NN622, GI262570, PNU182716, DRF552926, 2-[(5,7-dipropyl-3-trifluoromethyl-1,2-benzisoxazol-6-yl)oxy]-2-methylprop ionic acid (disclosed in U.S. Ser. No. 09/782,856), and 2(R)-7-(3-(2-chloro-4-(4-fluorophenoxy) phenoxy)propoxy)-2-ethylchromane-2-carboxylic acid (disclosed in U.S. Ser. Nos. 60/235,708 and 60/244,697). Thus, a method of treating or preventing diabetic retinopathy which comprises administering a therapeutically effective amount of a claimed compound in combination with a PPAR-.gamma. agonist is also within the scope of the present invention.

If formulated as a fixed dose, such combination products employ the compounds of this invention within the dosage range described below and the other pharmaceutically active agent(s) within its approved dosage range. Compounds of the instant invention may alternatively be used sequentially with known pharmaceutically acceptable agent(s) when a combination formulation is inappropriate.

The term "administration" and variants thereof (e.g., "administering" a compound) in reference to a compound of the invention means introducing the compound or a prodrug of the compound into the system of the animal in need of treatment. When a compound of the invention or prodrug thereof is provided in combination with one or more other active agents (e.g., a cytotoxic agent, etc.), "administration" and its variants are each understood to include concurrent and sequential introduction of the compound or prodrug thereof and other agents.

As used herein, the term "composition" is intended to encompass a product comprising the specified ingredients in the specified amounts, as well as any product which results, directly or indirectly, from combination of the specified ingredients in the specified amounts.

The term "therapeutically effective amount" as used herein means that amount of active compound or pharmaceutical agent that elicits the biological or medicinal response in a tissue, system, animal or human that is being sought by a researcher, veterinarian, medical doctor or other clinician.

The term "treating cancer" or "treatment of cancer" refers to administration to a mammal afflicted with a cancerous condition and refers to an effect that alleviates the cancerous condition by killing the cancerous cells, but also to an effect that results in the inhibition of growth and/or metastasis of the cancer.

The present invention also encompasses a pharmaceutical composition useful in the treatment of cancer, comprising the administration of a therapeutically effective amount of the compounds of this invention, with or without pharmaceutically acceptable carriers or diluents. Suitable compositions of this invention include aqueous solutions comprising compounds of this invention and pharmacologically acceptable carriers, e.g., saline, at a pH level, e.g., 7.4. The solutions may be introduced into a patient's bloodstream by local bolus injection.

When a compound according to this invention is administered into a human subject, the daily dosage will normally be determined by the prescribing physician with the dosage generally varying according to the age, weight, and response of the individual patient, as well as the severity of the patient's symptoms.

In one exemplary application, a suitable amount of compound is administered to a mammal undergoing treatment for cancer. Administration occurs in an amount between about 0.1 mg/kg of body weight to about 60 mg/kg of body weight per day, preferably of between 0.5 mg/kg of body weight to about 40 mg/kg of body weight per day.

The scope of the invetion therefore encompasses the use of the instantly claimed compounds in combination with a second compound selected from:

1) an estrogen receptor modulator,

2) an androgen receptor modulator,

3) retinoid receptor modulator,

4) a cytotoxic agent,

5) an antiproliferative agent,

6) a prenyl-protein transferase inhibitor,

7) an HMG-CoA reductase inhibitor,

8) an HIV protease inhibitor,

9) a reverse transcriptase inhibitor, and

10) another angiogenesis inhibitor.

Preferred angiogenesis inhibitors to be useed as the second compound are a tyrosine kinase inhibitor, an inhibitor of epidermal-derived growth factor, an inhibitor of fibroblast-derived growth factor, an inhibitor of platelet derived growth factor, an MMP (matrix metalloprotease) inhibitor, an integrin blocker, interferon-.alpha., interleukin-12, pentosan polysulfate, a cyclooxygenase inhibitor, carboxyamidotriazole, combretastatin A-4, squalamine, 6-O-chloroacetyl-carbonyl)-fumagillol, thalidomide, angiostatin, troponin-1, or an antibody to VEGF. Preferred estrogen receptor modulators are tamoxifen and raloxifene.

Also included in the scope of the claims is a method of treating cancer which comprises administering a therapeutically effective amount of a compound of Formula I in combination with radiation therapy and/or in combination with a compound selected from:

1) an estrogen receptor modulator,

2) an androgen receptor modulator,

3) retinoid receptor modulator,

4) a cytotoxic agent,

5) an antiproliferative agent,

6) a prenyl-protein transferase inhibitor,

7) an HMG-CoA reductase inhibitor,

8) an HIV protease inhibitor,

9) a reverse transcriptase inhibitor, and

10) another angiogenesis inhibitor.

And yet another embodiment of the invention is a method of treating cancer which comprises administering a therapeutically effective amount of a compound of Formual I in combination with paclitaxel or trastuzumab.

The invention further encompasses a method of treating or preventing cancer which comprises administering a therapeutically effective amount of a compound of Formula I in combination with a COX-2 inhibitor.

These and other aspects of the invention will be apparent from the teachings contained herein.

Definitions

The compounds of the present invention may have asymmetric centers, chiral axes, and chiral planes (as described in: E. L. Eliel and S. H. Wilen, Stereochemistry of Carbon Compounds, John Wiley & Sons, New York, 1994, pages 1119-1190), and occur as racemates, racemic mixtures, and as individual diastereomers, with all possible isomers and mixtures thereof, including optical isomers, being included in the present invention. In addition, the compounds disclosed herein may exist as tautomers and both tautomeric forms are intended to be encompassed by the scope of the invention, even though only one tautomeric structure is depicted. For example, any claim to compound A below is understood to include tautomeric structure B, and vice versa, as well as mixtures thereof. ##STR7##

When any variable (e.g. R.sup.4, R.sup.6, R.sup.6a, etc.) occurs more than one time in any constituent, its definition on each occurrence is independent at every other occurrence. Also, combinations of substituents and variables are permissible only if such combinations result in stable compounds. Lines drawn into the ring systems from substituents indicate that the indicated bond may be attached to any of the substitutable ring atoms. If the ring system is polycyclic, it is intended that the bond be attached to any of the suitable carbon atoms on the proximal ring only.

It is understood that substituents and substitution patterns on the compounds of the instant invention can be selected by one of ordinary skill in the art to provide compounds that are chemically stable and that can be readily synthesized by techniques known in the art, as well as those methods set forth below, from readily available starting materials. If a substituent is itself substituted with more than one group, it is understood that these multiple groups may be on the same carbon or on different carbons, so long as a stable structure results. The phrase "optionally substituted with one or more substituents" should be taken to be equivalent to the phrase "optionally substituted with at least one substituent" and in such cases the preferred embodiment will have from zero to three substituents.

As used herein, "alkyl" is intended to include both branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms. For example, C.sub.1 -C.sub.10, as in "C.sub.1 -C.sub.10 alkyl" is defined to include groups having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbons in a linear or branched arrangement. For example, "C.sub.1 -C.sub.10 alkyl" specifically includes methyl, ethyl, n-propyl, i-propyl, n-butyl, t-butyl, i-butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, and so on. The term "cycloalkyl" means a monocyclic saturated aliphatic hydrocarbon group having the specified number of carbon atoms. For example, "cycloalkyl" inlcudes cyclopropyl, methyl-cyclopropyl, 2,2-dimethyl-cyclobutyl, 2-ethyl-cyclopentyl, cyclohexyl, and so on.

"Alkoxy" represents either a cyclic or non-cyclic alkyl group of indicated number of carbon atoms attached through an oxygen bridge. "Alkoxy" therefore encompasses the definitions of alkyl and cycloalkyl above.

If no number of carbon atoms is specified, the term "alkenyl" refers to a non-aromatic hydrocarbon radical, straight, branched or cyclic, containing from 2 to 10 carbon atoms and at least one carbon to carbon double bond. Preferably one carbon to carbon double bond is present, and up to four non-aromatic carbon-carbon double bonds may be present. Thus, "C.sub.2 -C.sub.6 alkenyl" means an alkenyl radical having from 2 to 6 carbon atoms. Alkenyl groups include ethenyl, propenyl, butenyl, 2-methylbutenyl and cyclohexenyl. The straight, branched or cyclic portion of the alkenyl group may contain double bonds and may be substituted if a substituted alkenyl group is indicated.

The term "alkynyl" refers to a hydrocarbon radical straight, branched or cyclic, containing from 2 to 10 carbon atoms and at least one carbon to carbon triple bond. Up to three carbon-carbon triple bonds may be present. Thus, "C.sub.2 -C.sub.6 alkynyl" means an alkynyl radical having from 2 to 6 carbon atoms. Alkynyl groups include ethynyl, propynyl, butynyl, 3-methylbutynyl and so on. The straight, branched or cyclic portion of the alkynyl group may contain triple bonds and may be substituted if a substituted alkynyl group is indicated.

In certain instances, substituents may be defined with a range of carbons that includes zero, such as (C.sub.0 -C.sub.6)alkylene-aryl. If aryl is taken to be phenyl, this definition would include phenyl itself as well as --CH.sub.2 Ph, --CH.sub.2 CH.sub.2 Ph, CH(CH.sub.3)CH.sub.2 CH(CH.sub.3)Ph, and so on.

As used herein, "aryl" is intended to mean any stable monocyclic or bicyclic carbon ring of up to 7 atoms in each ring, wherein at least one ring is aromatic. Examples of such aryl elements include phenyl, naphthyl, tetrahydronaphthyl, indanyl, biphenyl, phenanthryl, anthryl or acenaphthyl. In cases where the aryl substituent is bicyclic and one ring is non-aromatic, it is understood that attachment is via the aromatic ring.

The term heteroaryl, as used herein, represents a stable monocyclic or bicyclic ring of up to 7 atoms in each ring, wherein at least one ring is aromatic and contains from 1 to 4 heteroatoms selected from the group consisting of O, N and S. Heteroaryl groups within the scope of this definition include but are not limited to: acridinyl, carbazolyl, cinnolinyl, quinoxalinyl, pyrrazolyl, indolyl, benzotriazolyl, furanyl, thienyl, benzothienyl, benzofuranyl, quinolinyl, isoquinolinyl, oxazolyl, isoxazolyl, indolyl, pyrazinyl, pyridazinyl, pyridinyl, pyriridinyl, pyrrolyl, tetrahydroquinoline. As with the definition of heterocycle below, "heteroaryl" is also understood to include the N-oxide derivative of any nitrogen-containing heteroaryl. In cases where the heteroaryl substituent is bicyclic and one ring is non-aromatic or contains no heteroatoms, it is understood that attachment is via the aromatic ring or via the heteroatom containing ring, respectively.

As appreciated by those of skill in the art, "halo" or "halogen" as used herein is intended to include chloro, fluoro, bromo and iodo. The term "heterocycle" or "heterocyclyl" as used herein is intended to mean a 5- to 10-membered aromatic or nonaromatic heterocycle containing from 1 to 4 heteroatoms selected from the group consisting of O, N and S, and includes bicyclic groups. "Heterocyclyl" therefore includes the above mentioned heteroaryls, as well as dihydro and tetrathydro analogs thereof. Further examples of "heterocyclyl" include, but are not limited to the following: benzoimidazolyl, benzofuranyl, benzofurazanyl, benzopyrazolyl, benzotriazolyl, benzothiophenyl, benzoxazolyl, carbazolyl, carbolinyl, cinnolinyl, furanyl, imidazolyl, indolinyl, indolyl, indolazinyl, indazolyl, isobenzofuranyl, isoindolyl, isoquinolyl, isothiazolyl, isoxazolyl, naphthpyridinyl, oxadiazolyl, oxazolyl, oxazoline, isoxazoline, oxetanyl, pyranyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridopyridinyl, pyridazinyl, pyridyl, pyrimidyl, pyrrolyl, quinazolinyl, quinolyl, quinoxalinyl, tetrahydropyranyl, tetrazolyl, tetrazolopyridyl, thiadiazolyl, thiazolyl, thienyl, triazolyl, azetidinyl, aziridinyl, 1,4-dioxanyl, hexahydroazepinyl, piperazinyl, piperidinyl, pyrrolidinyl, morpholinyl, thiomorpholinyl, dihydrobenzoimidazolyl, dihydrobenzofuranyl, dihydrobenzothiophenyl, dihydrobenzoxazolyl, dihydrofuranyl, dihydroimidazolyl, dihydroindolyl, dihydroisooxazolyl, dihydroisothiazolyl, dihydrooxadiazolyl, dihydrooxazolyl, dihydropyrazinyl, dihydropyrazolyl, dihydropyridinyl, dihydropyrimidinyl, dihydropyrrolyl, dihydroquinolinyl, dihydrotetrazolyl, dihydrothiadiazolyl, dihydrothiazolyl, dihydrothienyl, dihydrotriazolyl, dihydroazetidinyl, methylenedioxybenzoyl, tetrahydrofuranyl, and tetrahydrothienyl, and N-oxides thereof. Attachment of a heterocyclyl substituent can occur via a carbon atom or via a heteroatom.

The alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl and heterocyclyl substituents may be unsubstituted or unsubstituted, unless specifically defined otherwise. For example, a (C.sub.1 -C.sub.6)alkyl may be substituted with one, two or three substituents selected from OH, oxo, halogen, alkoxy, dialkylamino, or heterocyclyl, such as morpholinyl, piperidinyl, and so on. In this case, if one substituent is oxo and the other is OH, the following are included in the definition: --C.dbd.O)CH.sub.2 CH(OH)CH.sub.3, --(C.dbd.O)OH, --CH.sub.2 (OH)CH.sub.2 CH(O), and so on.

In certain instances, R.sup.7 and R.sup.8 are defined such that they can be taken together with the nitrogen to which they are attached to form a monocyclic or bicyclic heterocycle with 5-7 members in each ring and optionally containing, in addition to the nitrogen, one or two additional heteroatoms selected from N, O and S, said heterocycle optionally substituted with one or more substituents selected from R.sup.6a. Examples of the heterocycles that can thus be formed include, but are not limited to the following, keeping in mind that the heterocycle is optionally substituted with one or more substituents chosen from R.sup.6a : ##STR8##

Preferably R.sup.1 is H.

Also preferred is the definition of R.sup.2 and R.sup.3 as H.

Preferably R.sup.5 is H or F.

Preferably heterocyclyl is is selected from 2-azepinone, benzimidazolyl, 2-diazapinone, imidazolyl, 2-imidazolidinone, indolyl, isoquinolinyl, morpholinyl, piperidyl, piperazinyl, pyridyl, pyrrolidinyl, 2-piperidinone, 2-pyrimidinone, 2-pyrollidinone, quinolinyl, tetrahydrofuryl, tetrahydroisoquinolinyl, and thienyl.

Preferably R.sup.4a or R.sup.4b is H and the other is C.sub.1 -C.sub.6 alkyleneNR.sup.7 R.sup.8, said alkylene optionally substituted with oxo.

Preferably R.sup.7 and R.sup.8 are defined such that they are be taken together with the nitrogen to which they are attached to form a monocyclic 5-7 membered heterocycle and optionally containing, in addition to the nitrogen, one or two additional heteroatoms selected from N, O and S, and said heterocycle optionally substituted with one or more substituents selected from R.sup.6a.

The pharmaceutically acceptable salts of the compounds of this invention can be synthesized from the compounds of this invention which contain a basic or acidic moiety by conventional chemical methods. Generally, the salts of the basic compounds are prepared either by ion exchange chromatography or by reacting the free base with stoichiometric amounts or with an excess of the desired salt-forming inorganic or organic acid in a suitable solvent or various combinations of solvents. The pharmaceutically acceptable salts of the compounds of this invention include the conventional non-toxic salts of the compounds of this invention as formed, e.g., from non-toxic inorganic or organic acids. For example, such conventional non-toxic salts include those derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like: and the salts prepared from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, sulfanilic, 2-acetoxy-benzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isethionic, trifluoroacetic and the like. Similarly, the salts of the acidic compounds are formed by reactions with the appropriate inorganic or organic base.

The compounds of this invention may be prepared by employing reactions as shown in the following schemes, in addition to other standard manipulations that are known in the literature or exemplified in the experimental procedures. These schemes, therefore, are not limited by the compounds listed or by any particular substituents employed for illustrative purposes. Substituent numbering as shown in the schemes does not necessarily correlate to that used in the claims.

Schemes

As shown in Scheme A, the quinoline reagent A-2 can be synthesized by the general procedures taught in Marsais, F; Godard, A.; Queguiner, G. J. Heterocyclic Chem. 1989, 26, 1589-1594). Derivatives with varying substitution can be made by modifying this procedure and use of standard synthetic protocols known in the art. Also shown in Scheme 1 is the preparation of the indole intermediate A-5.

Scheme B illustrates one possible protocol for the coupling of the indole and quinolone intermediates to produce the desired compounds. Scheme C illustrates one possible synthetic route to the synthesis of a representative compound of the present invention, 3-(5-methoxy-1H-pyrrolo[2,3-c]pyridin-2-yl)-1H-quinolin-2-one, C-6. The starting chloro-compounds can be prepared according to the method taught by D. J. Pokoiny and W. W. Paudler in J. Org. Chem. 1972, 37, 3101. The selective fluorination of the indole substituent using 1-(chloromethyl)-4-fluoro-1,4-diazoniabicyclo[2.2.2]octane ditetrafluoroborate (Selectfluor.TM.) is exemplified in Scheme E. ##STR9## ##STR10## ##STR11## ##STR12## ##STR13## ##STR14##

Assays

The compounds of the instant invention described in the Examples were tested by the assays described below and were found to have kinase inhibitory activity. Other assays are known in the literature and could be readily performed by those of skill in the art. (see, for example, Dhanabal et al., Cancer Res. 59:189-197; Xin et al., J. Biol. Chem. 274:9116-9121; Sheu et al., Anticancer Res. 18:4435-4441; Ausprunk et al., Dev. Biol. 38:237-248; Gimbrone et al., J. Natl. Cancer Inst. 52:413-427; Nicosia et al., In Vitro 18:538-549.)

I. VEGF Receptor Kinase Assay

VEGF receptor kinase activity is measured by incorporation of radio-labeled phosphate into polyglutamic acid, tyrosine, 4:1 (pEY) substrate. The phosphorylated pEY product is trapped onto a filter membrane and the incorporation of radio-labeled phosphate quantified by scintillation counting.

Materials

VEGF Receptor Kinase

The intracellular tyrosine kinase domains of human KDR (Terman, B. I. et al. Oncogene (1991) vol. 6, pp. 1677-1683.) and Flt-1 (Shibuya, M. et al. Oncogene (1990) vol. 5, pp. 519-524) were cloned as glutathione S-transferase (GST) gene fusion proteins. This was accomplished by cloning the cytoplasmic domain of the KDR kinase as an in frame fusion at the carboxy terminus of the GST gene. Soluble recombinant GST-kinase domain fusion proteins were expressed in Spodoptera frugiperda (Sf21) insect cells (Invitrogen) using a baculovirus expression vector (pAcG2T, Pharmingen).

The other materials used and their compositions were as follows: Lysis buffer: 50 mM Tris pH 7.4, 0.5 M NaCl, 5 mM DTT, 1 mM EDTA, 0.5% triton X-100, 10% glycerol, 10 mg/mL of each leupeptin, pepstatin and aprotinin and 1 mM phenylmethylsulfonyl fluoride (all Sigma). Wash buffer: 50 mM Tris pH 7.4, 0.5 M NaCl, 5 mM DTT, 1 mM EDTA, 0.05% triton X-100, 10% glycerol, 10 mg/mL of each leupeptin, pepstatin and aprotinin and 1 mM phenylmethylsulfonyl fluoride. Dialysis buffer: 50 mM Tris pH 7.4, 0.5 M NaCl, 5 mM DTT, 1 mM EDTA, 0.05% triton X-100, 50% glycerol, 10 mg/mL of each leupeptin, pepstatin and aprotinin and 1 mM phenylmethylsuflonyl fluoride. 10.times. reaction buffer: 200 mM Tris, pH 7.4, 1.0 M NaCl, 50 mM MnCl.sub.2, 10 mM DTT and 5 mg/mL bovine serum albumin (Sigma). Enzyme dilution buffer: 50 mM Tris, pH 7.4, 0.1 M NaCl, 1 mM DTT, 10% glycerol, 100 mg/mL BSA. 10.times. Substrate: 750 .mu.g/mL poly (glutamic acid, tyrosine; 4:1) (Sigma). Stop solution: 30% trichloroacetic acid, 0.2 M sodium pyrophosphate (both Fisher). Wash solution: 15% trichloroacetic acid, 0.2 M sodium pyrophosphate. Filter plates: Millipore #MAFC NOB, GF/C glass fiber 96 well plate.

Method

A. Protein Purification

1. Sf21 cells were infected with recombinant virus at a multiplicity of infection of 5 virus particles/cell and grown at 27.degree. C. for 48 hours.

2. All steps were performed at 4.degree. C. Infected cells were harvested by centrifugation at 1000.times.g and lysed at 4.degree. C. for 30 minutes with 1/10 volume of lysis buffer followed by centrifugation at 100,000.times.g for 1 hour. The supernatant was then passed over a glutathione Sepharose column (Pharmacia) equilibrated in lysis buffer and washed with 5 volumes of the same buffer followed by 5 volumes of wash buffer. Recombinant GST-KDR protein was eluted with wash buffer/10 mM reduced glutathione (Sigma) and dialyzed against dialysis buffer.

B. VEGF Receptor Kinase Assay

1. Add 5 .mu.l of inhibitor or control to the assay in 50% DMSO.

2. Add 35 .mu.l of reaction mix containing 5 .mu.l of 10.times. reaction buffer, 5 .mu.l 25 mM ATP/10 .mu.Ci [.sup.33 P]ATP (Amersham), and 5 .mu.l 10.times. substrate.

3. Start the reaction by the addition of 10 .mu.l of KDR (25 nM) in enzyme dilution buffer.

4. Mix and incubate at room temperature for 15 minutes.

5. Stop by the addition of 50 .mu.l stop solution.

6. Incubate for 15 minutes at 4.degree. C.

7. Transfer a 90 .mu.l aliquot to filter plate.

8. Aspirate and wash 3 times with wash solution.

9. Add 30 .mu.l of scintillation cocktail, seal plate and count in a Wallac Microbeta scintillation counter.

II. Human Umbilical Vein Endothelial Cell Mitogenesis Assay

Human umbilical vein endothelial cells (HUVECs) in culture proliferate in response to VEGF treatment and can be used as an assay system to quantify the effects of KDR kinase inhibitors on VEGF stimulation. In the assay described, quiescent HUVEC monolayers are treated with vehicle or test compound 2 hours prior to addition of VEGF or basic fibroblast growth factor (bFGF). The mitogenic response to VEGF or bFGF is determined by measuring the incorporation of [.sup.3 H]thymidine into cellular DNA.

Materials HUVECs: HUVECs frozen as primary culture isolates are obtained from Clonetics Corp. Cells are maintained in Endothelial Growth Medium (EGM; Clonetics) and are used for mitogenic assays described in passages 3-7 below. Culture Plates: NUNCLON 96-well polystyrene tissue culture plates (NUNC #167008). Assay Medium: Dulbecco's modification of Eagle's medium containing 1 g/mL glucose (low-glucose DMEM; Mediatech) plus 10% (v/v) fetal bovine serum (Clonetics). Test Compounds: Working stocks of test compounds are diluted serially in 100% dimethylsulfoxide (DMSO) to 400-fold greater than their desired final concentrations. Final dilutions to 1.times. concentration are made directly into Assay Medium immediately prior to addition to cells. 10.times. Growth Factors: Solutions of human VEGF.sub.165 (500 ng/mL; R&D Systems) and bFGF (10 ng/mL; R&D Systems) are prepared in Assay Medium. 10.times. [.sup.3 H]Thymidine: [Methyl-.sup.3 H]thymidine (20 Ci/mmol; Dupont-NEN) is diluted to 80 .mu.Ci/mL in low-glucose DMEM. Cell Wash Medium: Hank's balanced salt solution (Mediatech) containing 1 mg/mL bovine serum albumin (Boehringer-Mannheim). Cell Lysis Solution: 1 N NaOH, 2% (w/v) Na.sub.2 CO.sub.3.

Method

1. HUVEC monolayers maintained in EGM are harvested by trypsinization and plated at a density of 4000 cells per 100 .mu.L Assay Medium per well in 96-well plates. Cells are growth-arrested for 24 hours at 37.degree. C. in a humidified atmosphere containing 5% CO.sub.2.

2. Growth-arrest medium is replaced by 100 .mu.L Assay Medium containing either vehicle (0.25% [v/v] DMSO) or the desired final concentration of test compound. All determinations are performed in triplicate. Cells are then incubated at 37.degree. C. with 5% CO.sub.2 for 2 hours to allow test compounds to enter cells.

3. After the 2-hour pretreatment period, cells are stimulated by addition of 10 .mu.L/well of either Assay Medium, 10.times.VEGF solution or 10.times.bFGF solution. Cells are then incubated at 37.degree. C. and 5% CO.sub.2.

4. After 24 hours in the presence of growth factors, 10.times. [.sup.3 H]thymidine (10 .mu.L/well) is added.

5. Three days after addition of [.sup.3 H]thymidine, medium is removed by aspiration, and cells are washed twice with Cell Wash Medium (400 .mu.L/well followed by 200 .mu.L/well). The washed, adherent cells are then solubilized by addition of Cell Lysis Solution (100 .mu.L/well) and warming to 37.degree. C. for 30 minutes. Cell lysates are transferred to 7-mL glass scintillation vials containing 150 .mu.L of water. Scintillation cocktail (5 mL/vial) is added, and cell-associated radioactivity is determined by liquid scintillation spectroscopy.

Based upon the foregoing assays the compounds of Formula I are inhibitors of VEGF and thus are useful for the inhibition of angiogenesis, such as in the treatment of ocular disease, e.g., diabetic retinopathy and in the treatment of cancers, e.g., solid tumors. The instant compounds inhibit VEGF-stimulated mitogenesis of human vascular endothelial cells in culture with IC.sub.50 values between 0.01-5.0 .mu.M. These compounds may also show selectivity over related tyrosine kinases (e.g., FGFR1 and the Src family; for relationship between Src kinases and VEGFR kinases, see Eliceiri et al., Molecular Cell, Vol. 4, pp. 915-924, December 1999).

III. FLT-1 Kinase Assay

Flt-1 was expressed as a GST fusion to the Flt-1 kinase domain and was expressed in baculovirus/insect cells. The following protocol was employed to assay compounds for Flt-1 kinase inhibitory activity: 1. Inhibitors were diluted to account for the final dilution in the assay, 1:20. 2. The appropriate amount of reaction mix was prepared at room temperature:

10.times. Buffer (20 mM Tris pH 7.4/0.1 M NaCl/1 mM DTT final)

0.1M MnCl.sub.2 (5 mM final)

pEY substrate (75 .mu.g/mL)

ATP/[.sup.33 P]ATP (2.5 .mu.M/1 .mu.Ci final)

BSA (500 .mu.g/mL final). 3. 5 .mu.L of the diluted inhibitor was added to the reaction mix. (Final volume of 5 .mu.L in 50% DMSO). To the positive control wells, blank DMSO (50%) was added. 4. 35 .mu.L of the reaction mix was added to each well of a 96 well plate. 5. Enzyme was diluted into enzyme dilution buffer (kept at 4.degree. C.). 6. 10 .mu.L of the diluted enzyme was added to each well and mix (5 nM final). To the negative control wells, 10 .mu.L 0.5 M EDTA was added per well instead (final 100 mM). 7. Incubation was then carried out at room temperature for 30 minutes. 8. Stopped by the addition of an equal volume (50 .mu.L) of 30% TCA/0.1M Na pyrophosphate. 9. Incubation was then carried out for 15 minutes to allow precipitation. 10. Transfered to Millipore filter plate. 11. Washed 3.times. with 15% TCA/0.1M Na pyrophosphate (125 .mu.L per wash). 12. Allowed to dry under vacuum for 2-3 minutes. 13. Dryed in hood for .about.20 minutes. 14. Assembled Wallac Millipore adapter and added 50 .mu.L of scintillant to each well and counted.

Indolinyl-isoquinolinone compounds which specifically inhibit, regulate and/or modulate the signal transduction of tyrosine kinases have been previously reported, see WO 01/29025, published 26 Apr. 2001, herein incorporated by reference. The present compounds, however, exhibit enhanced pharmaceutical properties, such as pharmacokinetics and side effect profiles

EXAMPLES

Examples provided are intended to assist in a further understanding of the invention. Particular materials employed, species and conditions are intended to be illustrative of the invention and not limiting of the reasonable scope thereof. ##STR15##

2-Chloro-3-iodo-quinoline (1-2)

A suspension of 3-(2-chloro)-quinolineboronic acid (1--1, 5.05 g, 24.3 mmol, 1 equiv, prepared by the method of Marsais, F; Godard, A.; Queguiner, G. J. Heterocyclic Chem. 1989, 26, 1589-1594) and N-iodosuccinimide (5.48 g, 24.4 mmol, 1.00 equiv) in acetonitrile (300 mL) was stirred at 23.degree. C. in the dark for 20 hours. The reaction mixture was concentrated to dryness, and the resulting yellow solid was partitioned between saturated aqueous sodium bicarbonate solution and dichloromethane. The organic layer was washed with water, then dried over magnesium sulfate and concentrated to give 2-chloro-3-iodo-quinoline as a pale yellow solid. .sup.1 H NMR (400 MHz, CDCl.sub.3) .delta. 8.67 (s, 1H), 7.99 (br d, 1H, J=8.4 Hz), 7.75 (br t, 1H, J=7.7 Hz), 7.72 (br d, 1H, J=7.8 Hz), 7.57 (br t, 1H, J=7.6 Hz).

3-Iodo-1H-quinolin-2-one (1-3)

The 2-chloro-3-iodoquinoline (1-2, 30.0 g) was weighed into a 250 mL flask and suspended in of 50% aqueous acetic acid (125 mL). The mixture was heated to 100.degree. C. and allowed to reflux for 16 hours to completion by TLC analysis of the crude reaction mixture. The mixture was allowed to cool to ambient temperature followed by dilution with 200 mL of water. The desired product was isolated by vacuum filtration followed by washing with water (50 mL). The water and traces of acetic acid were removed under vacuum for 5 hours to afford the desired quinolinone as a tan powder (1-3); .sup.1 H NMR (500 MHz, CDCl.sub.3) .delta. 12.13 (br s, 1H), 8.71 (s, 1H), 7.65 (d, 1H, J=7.5 Hz), 7.54 (m, 1H), 7.31 (d, 1H, J=8.0 Hz), 7.20 (m, 1H).

1-tert-butyl 6-methyl 1H-indole-1,6-dicarboxylate (1-5)

4-Dimethylaminopyridine (290 mg, 2.37 mmol, 0.10 equiv) was added to a solution of methyl 1H-indole-6-carboxylate (1-4, 4.16 g, 23.7 mmol), di-tert-butyldicarbonate (5.7 g, 26 mmol, 1.1 equiv) at 0.degree. C., and the resulting mixture was allowed to warm to room temperature over 1.5 hours. The mixture was partitioned between CH.sub.2 Cl.sub.2 (2.times.100 mL) and water (120 mL). The combined organics layers were dried over sodium sulfate and concentrated to afford 1-tert-butyl 6-methyl 1H-indole-1,6-dicarboxylate (1-5) as a colorless oil. .sup.1 H NMR (300 MHz, CDCl.sub.3) .delta. 8.88 (s, 1H), 7.94 (dd, 1H, J=8.2, 1.8 Hz), 7.75 (br d, 1H, J=3.7 Hz), 7.60 (d, 1H, J=8.2 Hz), 6.62 (dd, 1H, J=3.7, 1.5 Hz), 3.95 (s, 3H), 1.71 (s, 9H).

1-tert-butyl 6-methyl 2-(trimethylstannyl)-1H-indole-1,6-dicarboxylate (1-6)

A solution of lithium diisopropylamide (0.50 M, 72.6 mL, 36.2 mmol, 1.6 equiv) was added via cannula to a solution of 1-tert-butyl 6-methyl 1H-indole-1,6-dicarboxylate (1-5, 6.23 g, 22.6 mmol) in THF (100 mL) at -78.degree. C. The resulting mixture was stirred for 2 hours at -78.degree. C. before a solution of trimethyltin chloride in THF (2.3 M, 19.7 ml, 45.3 mmol, 2.00 equiv) was added. The resulting solution was warmed to 0.degree. C. for 15 minutes, then partitioned between EtOAc (2.times.150 mL) and aqueous half-saturated NH.sub.4 Cl solution (200 mL). The combined organics layers were dried over sodium sulfate and concentrated. The residue was purified by flash column chromatography (100% hexanes initially, grading to 20% EtOAc in hexanes) to afford 1-tert-butyl 6-methyl 2-(trimethylstannyl)-1H-indole-1,6-dicarboxylate (1-6) as an orange oil. .sup.1 H NMR (300 MHz, CDCl.sub.3) .delta. 8.74 (s, 1H), 7.88 (dd, 1H, J=8.2, 1.8 Hz), 7.53 (d, 1H, J=8.2 Hz), 6.77 (br s, 1H), 3.95 (s, 3H), 1.74 (s, 9H), 0.33 (s, 9H).

3-{6-[(4-methylpiperazin-1-yl)carbonyl]-1H-indol-2-yl}quinolin-2(1H)-one (1-7)

A solution of 1-tert-butyl 6-methyl 2-(trimethylstannyl)-1H-indole-1,6-dicarboxylate (1-6, 3.56 g, 8.12 mmol, 2.00 equiv) in dioxane (15 ml) was added in two equal portions over 2 hours to a solution of 3-iodo-1H-quinolin-2-one (5--5, 1.10 g, 4.06 mmol, 1 equiv), copper iodide (77 mg, 0.41 mmol, 0.10 equiv), and Pd(PPh.sub.3).sub.4 (234 mg, 0.202 mmol, 0.050 equiv) at 90.degree. C. The reaction mixture was then stirred for 24 hours, then partitioned between EtOAc (3.times.150 mL) and a 3:1 mixture of aqueous saturated NaHCO.sub.3 solution and brine (300 mL). The combined organic layers were washed with brine (250 mL), dried over sodium sulfate and concentrated. A solution of the residue in t-BuOH (50 mL) was treated with NaOH (1N, 4.06 mL, 4.06 mmol, 1 equiv) and heated to 50.degree. C. for 24 hours. The solvent was removed in vacuo, and residue was partitioned between ether (50 mL) and aqueous NaHCO.sub.3 solution (50 mL). The aqueous layer was acidified to pH 6 with aqueous 1N HCl solution, then extracted with EtOAc (3.times.130 mL). The combined organic layers were dried over Na.sub.2 SO.sub.4 concentrated. A solution of the residue (primarily 2-(2-oxo-1,2-dihydroquinolin-3-yl)-1H-indole-6-carboxylic acid, 30 mg), N-methylpiperazine (10 .mu.L, 0.12 mmol, 1.2 equiv), EDC (23 mg, 0.12 mmol, 1.2 equiv), HOAt (16 mg, 0.12 mmol, 1.2 equiv), and triethylamine (30 .mu.L, 0.25 mmol, 2.5 equiv) in DMF (10 mL) was stirred for 24 hours at ambient temperature. The reaction mixture was partitioned between EtOAc (3.times.60 mL) and water (100 mL), and the combined organic layers were dried over Na.sub.2 SO.sub.4, filtered and concentrated. The residual brown oil was purified via reverse phase LC (H.sub.2 O/CH.sub.3 CN gradient with 0.1% TFA present) to provide 3-{6-[(4-methylpiperazin-1-yl)carbonyl]-1H-indol-2-yl}quinolin-2(1H)-one (1-7). .sup.1 H NMR (300 MHz, DMSO) .delta. 12.25 (s, 1H), 11.86 (s, 1H), 8.63 (s, 1H), 7.77 (d, 1H, J=7.9 Hz), 7.69 (s, 1H), 7.64 (d, 1H, J=8.2 Hz), 7.56 (t, 1H, J=7.6 Hz), 7.38 (m, 2H), 7.27 (t, 1H, J=7.3 Hz), 7.13 (d, 1H, J=8.5 Hz), 4.30 (br s, 2H), 3.35 (br s, 4H), 3.17 (br s, 2H), 2.84 (br s, 3H).

The compounds in Table 1 below were prepared by simple modifications of the above procedure.

TABLE 1 1-8 ##STR16## N-methyl-2-(2-oxo-1,2- dihydroquinolin-3-yl)-N- pyrrolidin-3-yl-1H-indole- 6-carboxamide 1-9 ##STR17## 3-{4-[(4-methylpiperazin-1-yl) carbonyl]-1H-indol-2-yl}quinolin- 2(1H)-one 1-10 ##STR18## N-[2-(dimethylamino)ethyl]-2-(2- oxo-1,2-dihydroquinolin-3-yl)-1H- indole-4-carboxamide 1-11 ##STR19## N-methyl-2-(2-oxo-1,2- dihydroquinolin-3-yl)-N- pyrrolidin-3-yl-1H-indole-4- carboxamide

##STR20##

6-({[tert-butyl(dimethyl)silyl]oxy}methyl)-2-(2-oxo-1,2-dihydroquinolin-3-y l)-1H-indole-1-carboxylic acid tert-butyl ester (2-1)

3-Iodoquinolinone (800 mg, 2.95 mmol), the indole-boronic acid (622 mg, 0.5 equiv., 1.48 mmol), and lithium chloride (375 mg, 3 equiv., 8.85 mmol) were placed in a 25 mL flask equipped with a condenser. The mixture was diluted with 15 mL of dioxane and then degassed. The Pd(PPh.sub.3).sub.4 was added followed by Na.sub.2 CO.sub.3 solution (2 M, 5 mL). The reaction was degassed again and N.sub.2 was bubbled into the reaction mixture. The reaction was stirred in an atmosphere of nitrogen at 80.degree. C. The reaction was initially checked after stirring for 2 hours. Additional boronic acid (0.2 equiv.) was added every 30 minutes until most of the iodoqinolinone was consumed. The reaction mixture was then diluted with ethyl acetate (100 mL) and water (10 mL). The two layers were separated and the aqueous layer was extracted with ethyl acetate (2.times.50 mL). The combined organic phases were washed with brine, dried with Na.sub.2 SO.sub.4 and condensed under the reduced pressure. The crude product was chromatographed through flash silica gel with ethyl acetate/hexane (1:4) as eluent to give 2-1 as a yellow oil; .sup.1 H NMR (500 MHz, CDCl.sub.3) .delta. 12.52 (s, 1H), 8.08 (s, 1H), 7.89 (s, 1H), 7.44 (d, 1H, J=7.1 Hz), 7.40 (d, 1H, J=8.1 Hz), 7.32 (dd, 1H, J=1.2, 8.3 Hz), 7.23 (d, 1H, J=7.8 Hz), 7.13 (d, 1H, J=7.8 Hz)7.07 (t, 1H, J=7.1 Hz), 6.52 (s, 1H), 4.76 (s, 2H), 1.34 (s, 9H).

6-(carboxaldehyde)-2-(2-oxo-1,2-dihydroquinolin-3-yl)-1H-indole-1-carboxyli c acid tert-butyl ester (2--2)

6-({[tert-butyl(dimethyl)silyl]oxy}methyl)-2-(2-oxo-1,2-dihydroquinolin-3-y l)-1H-indole-1-carboxylic acid tert-butyl ester, 2-1, (1.35 g, 2.67 mmol) was added to a polypropylene container and dissolved in 150 mL of tetrahydrofuran. The solution was cooled to 0.degree. C. HF-pyridine was added, and the reaction stirred at 0.degree. C. for 20 minutes. An additional 2 mL of HF-pyridine was added, and the reaction was left to stir at room temperature for 1 hour. The reaction mixture was added to a well stirred solution of saturated NaHCO.sub.3 and ethyl acetate. The two layers were separated, and the aqueous layer was extracted with ethyl acetate. The combined organic extracts were dried with Na.sub.2 SO.sub.4, filtered and condensed to near dryness. The yellow solid was collected by vacuum filtration. The product was triturated with ethyl acetate twice, then was oxidized directly to the aldehyde. The crude alcohol (2.67 mmol) was suspended in 25 mL of CH.sub.2 Cl.sub.2. Manganese(IV) oxide (0.521 g, 6.0 mmol) was added, and the reaction mixture was refluxed for one hour. An additional 3 equivalents of Mn(IV)O.sub.2 was then added every hour until the starting alcohol was no longer visible on TLC (thin layer chromatography). The reaction was cooled to room temperature and filtered through a fritted funnel containing celite. The celite was washed several times with MeOH. The mother liquor was concentrated in vacuo. The yellow solid was triturated twice with methanol (10 mL), ethyl acetate (30 mL), and hexane (20 mL). The aldehyde, 2--2, was collected as a yellow solid. .sup.1 H NMR (500 MHz, DMSO-d.sub.6) .delta. 12.06 (s, 1H), 10.05 (s, 1H), 8.64 (s, 1H), 8.17 (s, 1H), 7.85 (m, 2H), 7.78 (d, 1H, J=7.9 Hz), 7.56 (m, 1H), 7.34 (d, 1H, J=8.2 Hz), 7.21 (t, 1H, J=8.2 Hz), 6.75 (s, 1H), 1.34 (s, 9H)

6-{[4-(methylsulfonyl)piperazin-1-yl]methyl}-2-(2-oxo-1,2-dihydroquinolin-3 -yl)-1H-indole-1-carboxylate (2-3)

2--2 (200 mg, 0.52 mmol) and amine (845 mg, 10 equiv., 5.15 mmol) were dissolved in 40 mL of DCE (dichloroethane). Acetic acid (2.0 mL) was added followed by sodium triacetoxyborohydride (327 mg, 3 equiv., 1.54 mmol). The hydride was added slowly over a period of 3 hours at room temperature. The reaction was left to stir for an additional 0.5 hours. The DCE was concentrated in vacuum to half its volume, diluted with with ethyl acetate and partitioned with saturated NaHCO.sub.3. The combined ethyl acetate layers were washed with water, brine, dried with Na.sub.2 SO.sub.4 and concentrated in vacuum to give the crude product, 2-3. The product was dissolved in warm DMF (2 mL) and solidified with MeOH (20 mL). The solid was collected by filtration and dried under the reduced pressure. .sup.1 H NMR (500 MHz, DMSO-d.sub.6) .delta. 12.06 (s, 1H), 8.13 (s, 1H), 8.09 (s, 1H), 7.73 (d, 1H, J=7.8 Hz), 7.57 (d, 1H, J=7.81 Hz), 7.52 (t, 1H, J=7.6 Hz), 7.34 (d, 1H, J=8.3 Hz), 7.21 (d, 2H, J=7.8 Hz), 6.75 (s, 1H), 3.62 (s, 2H), 3.16 (m, 4H), 2.87 (s, 3H), 2.48 (m, 4H), 1.35 (s, 9H).

3-(6-{[4-(methylsulfonyl)piperazin-1-yl]methyl}-1H-indol-2-yl)quinolin-2(1H )-one (2-4)

Boc-indole 2-3 (120 mg, 0.22 mmol) was slurried in 3 mL of CH.sub.2 Cl.sub.2, H.sub.2 O (10 .mu.L) and dimethylsulfide (10 .mu.L). TFA (3 mL) was added and the reaction was observed to become homogenous. The reaction was then stirred for 1.5 hours. The excess TFA/CH.sub.2 Cl.sub.2 was removed under reduced pressure. The residue was dissolved in DMSO and purified via reverse phase HPLC (95%-5% H.sub.2 O/AcCN). The pure fractions were combined in a separatory funnel and the pH adjusted to 11 by the addition of 0.5N NaOH. The mixture was then extracted with CH.sub.2 Cl.sub.2 /nBuOH (3:1, 2.times.100 mL). The organic layer was washed with brine, dried with Na.sub.2 SO.sub.4 and concentrated to give 2-4 as a yellow solid. .sup.1 H NMR (500 MHz, DMSO-d.sub.6) .delta. 12.06 (s, 1H), 11.43 (s, 1H), 8.52 (s, 1H), 7.73 (d, 1H, J=7.8 Hz), 7.51 (d, 2H, J=7.3 Hz), 7.44 (bs, 1H), 7.37 (d, 1H, J=8.1 Hz), 7.31 (bs, 1H), 7.24 (t, 1H, J=7.3 Hz), 6.98 (d, 1H, J=7.6 Hz), 3.62 (s, 2H), 3.12 (m, 4H), 2.87 (s, 3H), 2.48 (m, 4H).

The compounds in Table 2 below were prepared by simple modifications of the procedure above:

TABLE 2 2-5 ##STR21## 4-[2-(2-Oxo-1,2-dihydro- quinolin-3-yl)-1H-indol-6- ylmethyl]-piperazine-1- carboxylic acid methylamide 2-6 ##STR22## 3-{4-[2-(2-Oxo-1,2-dihydro- quinolin-3-yl)-1H-indol-6-yl methyl]-piperazin-1-yl}1-butyric acid 2-7 ##STR23## 3-[4-(4-Methanesulfonyl- piperazin-1-ylmethyl)-1H-indol- 2-yl]-1H-quinolin-2-one 2-8 ##STR24## 3-{4-[2-(2-Oxo-1,2-dihydro- quinolin-3-yl)-1H-indol-4- ylmethyl]-piperazin-1-yl}- butyric acid

##STR25##

3-[3-Fluoro-5-(4-methanesulfonyl-piperazin-1-ylmethyl)-1H-indol-2-yl]-1H-qu inolin-2-one 3--3

3-(5-{[4-(methylsulfonyl)piperazin-1-yl]methyl}-1H-indol-2-yl)quinolin-2(1H )-one (3-1, 60 mg, 1.37 mmol) was dissolved in 5 mL of anhydrous dimethylsolfoxide and diluted with 5 mL of anhydrous acetonitrile. The solution was cooled to 0.degree. C., and the fluorinating reagent, 1-(chloromethyl)-4-fluoro-1,4-diazoniabicyclo[2.2.2]octane ditetrafluoroborate, 3-2 (0.25 equiv., 12 mg, 0.03 mmol) was added. After addition, the ice bath was removed, and the reaction stirred at room temperature for 30 minutes. 3-2 was continually added in this fashion until half of the starting material was consumed. At which point, the reaction was diluted with ethyl acetate (150 mL) and partitioned with water (25 mL). The two layers were separated, and the organic layer was washed several times with water. The organic layer was condensed under vacuum. The crude residue was purified by reverse phase HPLC (5-95% ACN/H.sub.2 O-40 minutes). The pure fractions were combined and volatiles were removed under reduced pressure. The resulting solution was partitioned with ethyl acetate (200 mL) and washed with 1 N NaOH (15 mL) and brine. The organic layer was dried with Na.sub.2 SO.sub.4, filtered and condensed under vacuum. The resulting pure product was isolated as yellow solid; .sup.1 H NMR (500 MHz, DMSO-d.sub.6) .delta. 12.3 (s, 1H), 11.3 (s, 1H), 8.47 (s, 1H), 7.83 (d, 1H, J=7.8 Hz), 7.58-7.52 (m, 2H), 7.46 (s, 1H), 7.39 (d, 1H, J=8.3 Hz), 7.26 (t, 1H, J=7.8 Hz), 7.13 (d, 1H, J=8.5 Hz), 3.60 (s, H), 3.11 (m, 4H), 2.87 (s, 3H), 2.48 (m, 4H).

The following compounds may be made by following the protocol above by substituting the appropriate indolinyl isoquinolinone for 3-1.

TABLE 3 3-4 ##STR26## 3-[3-Fluoro-6-(4-methanesulfonyl- piperazin-1-ylmethyl)-1H-indol-2-yl]- 1H-quinolin-2-one 3-5 ##STR27## 4-[2-(3-Fluoro-2-oxo-1,2- dihydroquinolin-3-yl)-1H-indol-6- ylmethyl]-piperazine-1-carboxylic acid methylamide 3-6 ##STR28## 3-{4-[3-Fluoro-2-(2-oxo-1,2-dihydro- quinolin-3-yl)-1H-indol-6-ylmethyl]- piperazin-1-yl}-butyric acid 3-7 ##STR29## 3-[3-Fluoro-4-(4-methanesulfonyl- piperazin-1-ylmethyl)-1H-indol-2-yl]- 1H-quinolin-2-one 3-8 ##STR30## 3-{4-[3-Fluoro-2-(2-oxo-1,2-dihydro- quinolin-3-yl)-1H-indol-4-ylmethyl]- piperazin-1-yl}-butyric acid