TW201217361A - Method of treating abnormal cell growth - Google Patents

Abstract

The present invention relates to the use of c-Met/HGFR and VEGF inhibitors for treating abnormal cell growth in mammals. In particular, the invention provides methods of treating mammals suffering from cancer. In particular the invention provides methods of treating mammals suffering from metastatic cancer. In particular the invention provides methods of inhibiting metastatic cancer. In particular the invention relates to combination treatment using Sunitinib and crizotinib or PF-04217903.

Description

201217361 VI. Description of the Invention: [Technical Field of the Invention] The present invention relates to the use of c-Met/HGFR and VEGF inhibitors for the treatment of abnormal cell growth in mammals. In particular, the invention provides methods of treating a mammal suffering from cancer. In particular, the invention provides methods of treating a mammal suffering from metastatic cancer. In particular, the invention provides a method of inhibiting metastatic cancer. The present application claims the benefit of U.S. Provisional Application Serial No. 61/387,934, filed on Sep. 29, 2011, the content of which is hereby incorporated by reference in its entirety. [Prior Art] The negative effect of the VEGF (vascular endothelial growth factor) signaling pathway has been shown to play an important role in physiological and pathological angiogenesis. Therefore, the FDA recently approved bevacizumab (an anti-VEGF monoclonal antibody), which has opened a new era in cancer therapy (Ferrara N et al., Angiogenesis as a therapeutic target. Nature 2005; 438 (7070) : 967-74). In addition to blocking ligands such as bevacizumab, several other RTKIs (receptor tyrosine kinase inhibitors) that target the VEGF pathway (such as sunitinib and sorafenib) Approved by the FDA or at various stages of development (Ivy SP et al., An overview of small-molecule inhibitors of VEGFR signaling. Nat Rev Clin Oncol 2009; 6 (10): 569-79). However, despite the efficacy and survival benefits demonstrated in multiple therapeutic settings, most patients will eventually exhibit disease progression. The molecular and cellular mechanisms responsible for this non-reactivity are being extensively studied. From the point of view of the generation of blood vessels 158393.doc 201217361, several studies have shown that tumor cells and stroma (mainly including fibroblasts, outer cells, spinal cells and non-tumor compartments of mesenchymal stem cells) are activated by alternative angiogenic factors. To promote the development of resistance to angiogenic therapies such as FGF-2 (Casogas Ο Ο et al, Drug resistance by evasion of antiangiogenic targeting of VEGF signaling in late-stage Pancreatic islet tumors. Cancer cell 2005;8(4):299-309)), Bv8 (Variegata) (Shojaei F et al, Bv8 regulates myeloid-cell-dependent tumour angiogenesis. Nature 2007; 450 (7171): 825-31)) and PDGF (Fikcher plate growth factor (Fischer C et al, Anti-PIGF inhibits growth of VEGF (R)-inhibitor-resistant tumors without affecting healthy vessels. Cell 2007; 131 (3): 463- 75)).

HGF (hepatocyte growth factor/dispersion factor)/c_Met pathway is known to play an important role in different developmental stages and in tumor formation (You WK et al., The hepatocyte growth factor/c-Met signaling pathway as a therapeutic target to inhibit Angiogenesis. BMB reports 2008; 41 (12): 833-9). When HGF is enriched in mesenchymal cells, c-Met is also highly expressed in epithelial cells. It has also been found that c_Met is expressed in several other cell types including endothelial cells, nerve cells, cumulus cells and ectodermal cells. The expression of c-Met and HGF has been identified in several cancer types (such as bladder cancer, breast cancer, gastric cancer, colon cancer, and kidney cancer) (Peruzzi B et al., arging the c-Met signaling pathway in cancer. Clin 158393. Doc 201217361

Cancer Res 2006; 12(12): 3657-60). It has been suggested that activation of c-Met causes tumor cell proliferation, survival, and invasiveness via signaling pathways (such as PIK3/Akt, Src, STAT3, and Ras/Mek) (Comoglio PM et al., Drug development of MET inhibitors: targeting Oncogene addiction and expedience. Nature reviews 2008; 7(6): 504-16; Maulik G et al, Role of the hepatocyte growth factor receptor, c-Met, in oncogenesis and potential for therapeutic inhibition. Cytokine & growth factor reviews 2002 13(1): 41-59). It has been shown that c-Met activation induces tumor blood and tube formation in vascular structures primarily by inducing endothelial cell proliferation, migration and survival via a paracrine HGF (Birchmeier C et al, Met, metastasis, motility and more. Nat Rev Mol Cell Biol 2003;4(12):915-25) »HGF has also been shown to upregulate VEGF (a key angiogenesis inducer) and down-regulate thrombospondin-1(1)11>01111303卩011(1丨11- 1) (an effective angiogenesis inhibitor) is shown to promote jk tube formation (Zhang YW et al, Hepatocyte growth factor/scatter factor mediates angiogenesis through positive VEGF and negative thrombospondin 1 regulation. PNAS 2003; 100(22): 12718- twenty three). Similar to VEGF, the expression of c-Met and HGF is induced by HIF-Ια (hypoxia-inducible factor), which further promotes c-Met/HGF to promote angiogenesis, cell survival and invasion under adverse microenvironment conditions. Role (Pennacchietti S et al., Hypoxia promotes invasive growth by transcriptional activation of the met protooncogene. Cancer cell 2003; 3(4): 347-61; Wang GL et al., Characterization of 158393.doc 201217361 hypoxia-inducible factor 1 and regulation J-Biol-Chem 1993 268(29): 21513-8) o Consistent with these observations, clinical findings indicate that activation of the c-Met pathway is a poor prognostic factor in cancer patients (Abounader R et al, Scatter factor/hepatocyte growth factor in brain tumor growth and angiogenesis. Neuro-oncology 2005;7(4):436-51; Garcia S et al, Poor prognosis in breast carcinomas correlates with increased expression of targetable CD 146 and c-Met and with proteomic basal-like phenotype. Human pathology 2007; 38(6): 830-41 ; Peghini PL et al., Overexpressi On of epidermal growth factor and hepatocyte growth factor receptors in a proportion of gastrinomas correlates with aggressive growth and lower curability. Clin Cancer Res 2002; 8(7): 2273-85). Recent studies have indicated that HGF is highly expressed in bone marrow CD11b+Grl+ cells isolated from anti-VEGF therapeutically resistant tumors (Shojaei F et al., Tumor refractoriness to anti-VEGF treatment is mediated by CD 1 lb + Grl+ myeloid cells. Nature Biotechnology 2007; 25(8): 911-20). However, a sufficient understanding of the role of HGF in tumor angiogenesis and especially in mediating resistance to VEGF inhibitors remains to be determined. Agents that produce tumor angiogenesis, particularly VEGF inhibitors, have provided benefits to cancer patients. Similar to other anticancer agents, tumor recurrence has become one of the major challenges in patients treated with angiogenesis inhibitors. Recent reports of preclinical ploughs indicate that activation of the angiogenic pathway is one of the mechanisms of alternative 158393.doc 201217361 angiogenesis pathway in anti-VEGF-treated tumors (Bergers G et al., Modes of resistance to anti- Angiogenic therapy. Nat Rev Cancer 2008; 8(8): 592-603). In addition, two recent independent studies have shown that anti-angiogenic therapy may induce tumor progression and metastasis due to increased tumor cell invasiveness (Ebos JM et al., Accelerated metastasis after short-term treatment with a potent inhibitor of tumor angiogenesis. Cell 2009;15(3): 232-9; Paez-Ribes M et al, Antiangiogenic therapy elicits malignant progression of tumors to increased local invasion and distant metastasis. Cancer cell 2009; 15(3): 220-3 1). Collectively, these observations have highlighted the need to optimize the application of such agents in a clinical setting. SUMMARY OF THE INVENTION The embodiments of the invention described below can be combined with any of the other embodiments of the invention described herein, but such other embodiments are inconsistent with the embodiments in which they are combined. In one embodiment, the invention provides a method of treating cancer in a mammal in need thereof, comprising the step of administering a therapeutically effective amount of a VEGF inhibitor and a therapeutically effective amount of a c-Met/HGFR inhibitor. In certain embodiments, the mammal is a human. In certain embodiments, the cancer is selected from the group consisting of colon cancer, non-small cell lung cancer, melanoma, renal cancer, hepatocellular carcinoma, glioblastoma multiforme, and pancreatic neuroendocrine tumor. In certain embodiments, the VEGF inhibitor is selected from the group consisting of sunitinib, SU-14813, PF-337210, bevacizumab, and axi 158393.doc 201217361 axitinib. In certain embodiments, the VEGF inhibitor is sunitinib. In certain embodiments, the VEGF inhibitor is axitinib. In certain embodiments, the VEGF inhibitor is bevacizumab. In certain embodiments, the VEGF inhibitor is SU-14813. In certain embodiments, the VEGF inhibitor is PF-3 3 7210. In certain embodiments, the c-Met/HGFR inhibitor is chromatinib or PF-04217903. In certain embodiments, the c-Met/HGFR inhibitor is klotinib. In certain embodiments, the c-Met/HGFR inhibitor is PF-04217903. In certain embodiments, the VEGF inhibitor is sunitinib and the c-Met/HGFR inhibitor is klotinib. In certain embodiments, the VEGF inhibitor is sunitinib and the c-Met/HGFR inhibitor is PF-04217903. In certain embodiments, the VEGF inhibitor is axitinib and the c-Met/HGFR inhibitor is klotinib. In certain embodiments, the VEGF inhibitor is axitinib and the c-Met/HGFR inhibitor is PF-04217903 » In certain embodiments, the VEGF inhibitor is bevacizumab and c-Met/HGFR inhibition The agent is klotinib. In certain embodiments, the VEGF inhibitor is bevacizumab and the c-Met/HGFR inhibitor is PF-04217903. In certain embodiments, the VEGF inhibitor is SU-14813 and the c-Met/HGFR inhibitor is chromatinib. In certain embodiments, the VEGF inhibitor is SU-14813 and the c-Met/HGFR inhibitor is PF-04217903. In certain embodiments, the VEGF inhibitor is PF-337210 and the c-Met/HGFR inhibitor is 克lotinib. In certain embodiments, the VEGF inhibitor is PF-337210 and the c-Met/HGFR inhibitor is PF-04217903.

In another embodiment, the invention provides a pharmaceutical composition comprising a VEGF 158393.doc 201217361 inhibitor and a c-Met/HGFR inhibitor in a combination that is therapeutically effective in treating cancer in a mammal. In another embodiment, the invention provides a method of treating cancer in a mammal in need thereof, comprising the step of administering a pharmaceutical composition of the invention. In certain embodiments, the cancer is selected from the group consisting of colon cancer, non-small cell lung cancer, melanoma, renal cancer, hepatocellular carcinoma, glioblastoma multiforme, and pancreatic neuroendocrine tumor. In certain embodiments, the VEGF inhibitor is selected from the group consisting of sunitinib, SU-14813, PF-337210, bevacizumab, and axitinib. In certain embodiments, the VEGF inhibitor is sunitinib. In certain embodiments, the VEGF inhibitor is axitinib. In certain embodiments, the VEGF inhibitor is bevacizumab. In certain embodiments, the VEGF inhibitor is SU-14813. In certain embodiments, the VEGF inhibitor is PF-337210. In certain embodiments, the c-Met/HGFR inhibitor is chromatinib or PF-04217903. In certain embodiments, the c-Met/HGFR inhibitor is klotinib. In certain embodiments, the c-Met/HGFR inhibitor is PF-042 17903. In certain embodiments, the VEGF inhibitor is sunitinib and the c-Met/HGFR inhibitor is klotinib. In certain embodiments, the VEGF inhibitor is sunitinib and the c-Met/HGFR inhibitor is PF-04217903. In certain embodiments, the VEGF inhibitor is axitinib and the c-Met/HGFR inhibitor is klotinib. In certain embodiments, the VEGF inhibitor is axitinib and the c-Met/HGFR inhibitor is PF-04217903. In certain embodiments, the VEGF inhibitor is bevacizumab and the c-Met/HGFR inhibitor is klotinib. In certain embodiments, the VEGF inhibitor is bevacizumab and the c-Met/HGFR inhibitor is PF-04217903. In certain embodiments, the VEGF inhibitor is SU-14813 and the c-Met/HGFR inhibitor is 158393.doc 201217361 卓tinib. In certain embodiments, the VEGF inhibitor is SU-14813 and the c-Met/HGFR inhibitor is PF-04217903. In certain embodiments, the VEGF inhibitor is PF-337210 and the c-Met/HGFR inhibitor is Klotinib. In certain embodiments, the VEGF inhibitor is PF-337210 and the c-Met/HGFR inhibitor is PF-04217903. In another embodiment, the invention provides a method of inhibiting the production of blood vessels in a mammal comprising the step of administering a therapeutically effective amount of a VEGF inhibitor and a c-Met/HGFR inhibitor. In certain embodiments, the mammal is a human. In certain embodiments, the cancer is selected from the group consisting of colon cancer, non-small cell lung cancer, melanoma, renal cancer, hepatocellular carcinoma, glioblastoma multiforme, and pancreatic neuroendocrine tumor. In certain embodiments, the VEGF inhibitor is selected from the group consisting of sunitinib, 31-14813, PF-337210, bevacizumab, and axitinib. In certain embodiments, the VEGF inhibitor is sunitinib. In certain embodiments, the VEGF inhibitor is axitinib. In certain embodiments, the VEGF inhibitor is bevacizumab. In certain embodiments, the VEGF inhibitor is SU-14813. In certain embodiments, the VEGF inhibitor is PF-337210. In certain embodiments, the c-Met/HGFR inhibitor is klotinib or PF-04217903. In certain embodiments, the c-Met/HGFR inhibitor is klotinib. In certain embodiments, the c-Met/HGFR inhibitor is PF-04217903. In certain embodiments, the VEGF inhibitor is sunitinib and the c-Met/HGFR inhibitor is chromatinib. In certain embodiments, the VEGF inhibitor is sunitinib and the c-Met/HGFR inhibitor is PF-04217903. In certain embodiments, the VEGF inhibitor is axitinib and the c-Met/HGFR inhibitor is ketamine 158393.doc •10·201217361 尼. In certain embodiments, the VEGF# formulation is axitinib and the c Met/HGFR inhibitor is pF_G42179()3. In certain embodiments, the VEGF inhibition is bevacizumab and the e_Met/HGFR^ formulation is kovoutinib. In certain embodiments, the VEGF inhibitor is bevacizumab and the Met/HGFR inhibitor is pF_G42179()3. In certain embodiments, the VEGF inhibitor is SU_14813 and the cardio-inhibitor is klodinib. In certain embodiments, the VEGF inhibitor is su_i48i3 and the Met/HGFR inhibitor is PF_G42179G3. In certain embodiments, the VEGF inhibitor is 卯_33721〇 and the c_Met/HGFR inhibitor is 克lotinib. In certain embodiments, the VEGF inhibitor is ρρ_3372ι〇 and

The Met/HGFR inhibitor is pf-04217903. In another embodiment, the invention provides a method of inhibiting angiogenesis in a mammal having therapeutic needs, comprising the step of administering a pharmaceutical composition of the invention. In certain embodiments, the mammal is a human. In certain embodiments, the cancer is selected from the group consisting of colon cancer, non-small cell lung cancer 'melanoma, kidney cancer, hepatocellular carcinoma, glioblastoma multiforme, and pancreatic neuroendocrine tumor. In certain embodiments, the VEGF inhibitor is selected from the group consisting of sunitinib, su_14813, pF_337210, bevacizumab, and axitinib. In certain embodiments, the vegf inhibitor is sunitinib. In certain embodiments, the vegf inhibitor is axitinib. In certain embodiments, the VEGF inhibitor is bevacizumab. In certain embodiments, the VEGF inhibitor is SU_14813. In certain embodiments the 'VEGF inhibitor is PF-337210. In certain embodiments, the c_Met/HGFR inhibitor is klotinib or pF_042179〇3. In some embodiments, I58393.doc • 11. 201217361, the c-Met/HGFR inhibitor is klotinib. In certain embodiments, the c-Met/HGFR inhibitor is PF-04217903. In certain embodiments, the VEGF inhibitor is sunitinib and the c-Met/HGFR inhibitor is klotinib. In certain embodiments, the VEGF inhibitor is sunitinib and the c-Met/HGFR inhibitor is PF-04217903. In certain embodiments, the VEGF inhibitor is axitinib and the c-Met/HGFR inhibitor is krodinib. In certain embodiments, the VEGF inhibitor is axitinib and the c-Met/HGFR inhibitor is PF-04217903. In certain embodiments, the VEGF inhibitor is bevacizumab and the c-Met/HGFR inhibitor is krodinib. In certain embodiments, the VEGF inhibitor is bevacizumab and the c-Met/HGFR inhibitor is PF-〇42l7903. In certain embodiments, the VEGF inhibitor is SU-14813 and the c-Met/HGFR inhibitor is 克lotinib. In certain embodiments, the VEGF inhibitor is SU-14813 and the c-Met/HGFR inhibitor is PF-04217903. In certain embodiments, the VEGF inhibitor is PF-33 7210 and the c-Met/HGFR inhibitor is 克lotinib. In certain embodiments, the VEGF inhibitor is PF-337210 and the c-Met/HGFR inhibitor is PF-04217903. In another embodiment, the invention provides a method of treating a metastatic cancer in a mammal in need thereof, comprising the step of administering a therapeutically effective amount of a VEGF inhibitor and a c-Met/HGFR inhibitor. In certain embodiments, the mammal is a human. In certain embodiments, the cancer is selected from the group consisting of colon cancer, non-small cell lung cancer, melanoma, renal cancer, hepatocellular carcinoma, glioblastoma multiforme, and pancreatic neuroendocrine tumor. In certain embodiments, the VEGF inhibitor is selected from the group consisting of: 158393.doc • 12· 201217361 Sunitinib, SU-14813, PF-337210, bevacizumab, and axitinib. In certain embodiments, the VEGF inhibitor is sunitinib. In certain embodiments the 'VEGF inhibitor is axitinib. In certain embodiments, the VEGF inhibitor is bevacizumab. In certain embodiments, the VEGF inhibitor is SU-14813. In certain embodiments, the VEGF inhibitor is PF-337210. In certain embodiments, the C-Met/HGFR inhibitor is krodinib or PF-04217903. In certain embodiments, the C-Met/HGFR inhibitor is klotinib. In certain embodiments, the c-Met/HGFR inhibitor is PF-04217903. In certain embodiments, the VEGF inhibitor is sunitinib and the c-Met/HGFR inhibitor is klotinib. In certain embodiments, the VEGF inhibitor is sunitinib and the c-Met/HGFR inhibitor is PF-04217903. In certain embodiments, the VEGF inhibitor is axitinib and the c-Met/HGFR inhibitor is klotinib. In certain embodiments, the VEGF inhibitor is axitinib and the c-Met/HGFR inhibitor is PF-04217903. In certain embodiments, the VEGF inhibitor is bevacizumab and the c-Met/HGFR inhibitor is chromatinib. In certain embodiments, the VEGF inhibitor is bevacizumab and the c-Met/HGFR inhibitor is PF-04217903. In certain embodiments, the VEGF inhibitor is SU-14813 and the c-Met/HGFR inhibitor is 克lotinib. In certain embodiments, the VEGF inhibitor is SU-14813 and the c-Met/HGFR inhibitor is PF-04217903. In certain embodiments, the VEGF inhibitor is PF-337210 and the c-Met/HGFR inhibitor is 克lotinib. In certain embodiments, the VEGF inhibitor is PF-337210 and the c-Met/HGFR inhibitor is PF-04217903. In another embodiment, the invention provides a method of inhibiting metastasis of a 158393.doc-13-201217361 milk animal having a therapeutic need comprising administering a therapeutically effective amount of a VEGF inhibitor and a c-Met/HGFR inhibitor A step of. In certain embodiments, the transfer occurs in a lymph node. In certain embodiments, the metastasis occurs in the colon. In certain embodiments, the mammal is a human. In certain embodiments, the cancer is selected from the group consisting of colon cancer, non-small cell lung cancer, melanoma, renal cancer, hepatocellular carcinoma, glioblastoma multiforme, and splanchnic neuroendocrine tumor. In certain embodiments, the VEGF inhibitor is selected from the group consisting of sunitinib, SU-14813, PF-3 3 7210, bevacizumab, and axitinib. In certain embodiments, the VEGF inhibitor is sunitinib. In certain embodiments, the VEGF inhibitor is axitinib. In certain embodiments, the VEGF inhibitor is bevacizumab. In certain embodiments, the VEGF inhibitor is SU-14813. In certain embodiments, the VEGF inhibitor is PF-337210. In certain embodiments, the c-Met/HGFR inhibitor is klotinib or PF-042 17903. In certain embodiments, the c-Met/HGFR inhibitor is klotinib. In certain embodiments, the c-Met/HGFR inhibitor is PF-04217903. In certain embodiments, the VEGF inhibitor is sunitinib and the c-Met/HGFR inhibitor is klotinib. In certain embodiments, the VEGF inhibitor is sunitinib and the c-Met/HGFR inhibitor is PF-04217903. In certain embodiments, the VEGF inhibitor is axitinib and the c-Met/HGFR inhibitor is krodinib. In certain embodiments, the VEGF inhibitor is axitinib and the c-Met/HGFR inhibitor is PF-04217903 » In certain embodiments, the VEGF inhibitor is bevacizumab and c-Met/HGFR inhibition The agent is klotinib. In certain embodiments, the VEGF inhibitor is bevacizumab and c-158393.doc -14-201217361

The Met/HGFR inhibitor is pF-〇4217903. In certain embodiments, the VEGF inhibitor is SU-14813 and the c-Met/HGFR inhibitor is 克lotinib. In certain embodiments, the VEGF inhibitor is su_14813 and the e-Met/HGFR inhibitor is PF_042179〇3. In certain embodiments, the VEGF inhibitor is 卩? -337210 and the c_Met/HGFR inhibitor is klotinib. In certain embodiments, the VEGF inhibitor is c_

The Met/HGFR inhibitor is pf_〇42 1 7903. [Embodiment] Definition Unless otherwise indicated, the term "abnormal cell growth" as used herein refers to cell growth that is not governed by a normal regulatory mechanism (e.g., loss of contact inhibition). The term "treatment/treatment" as used herein, unless otherwise indicated, means that the condition or condition to which the term applies or the symptoms of a plurality of such conditions or conditions are inhibited or the progression thereof is inhibited. The term "treatment" as used herein, unless otherwise indicated, refers to the therapeutic/treatment effect of "treatment, treatment" as defined immediately above. The term "angiogenesis inhibitor" as used herein includes any substance that inhibits new (ie, angiogenesis). An angiogenesis inhibitor can be: by binding/inhibiting VEGF, inhibiting coffee, inhibiting endothelial cells/cells, cells, migration and survival, inducing endothelial cells, dying, and activating immune system A substance that stimulates the formation of angiogenesis inhibitors, inhibits membrane degradation, and the like. The term "metastasis" as used herein means that a disease (here a cancer or not 158393.doc 201217361::dimer growth) spreads from one organ or part to another non-adjacent "or P knife." The 'transfer used can be a local metastasis, a certain origin of the pot = the cancer cells of the primary tumor gain the ability to penetrate and soak the surrounding hanging tissue in a local area' to form a new tumor (sometimes called a "sub-tumor" As used herein, 'metastasis can also be lymphatic spread, and some of the cancer cells derived from the primary tumor can circulate in the bloodstream after obtaining the ability to penetrate the lymph node wall or lymphatic vessel wall (sometimes called It is a "circulating tumor cell" to other parts of the body and tissues. As used herein, metastasis can also be a blood-borne spread, in which some cancer cells that originate from the primary tumor circulate in the bloodstream after being able to penetrate the vessel wall (sometimes (iv) "circulating tumor cells ") to other parts of the body and tissues. The transfer process can be any process known to involve the proliferation of cancer cells in another part of the body, another organ or another tissue. By way of example, after the tumor cells are from the primary tumor: after the other site is at rest, the cells can then penetrate the blood vessel or wall, continue to expand and eventually form another clinically detectable tumor. When tumor cells metastasize, 'new tumors are called secondary or metastatic tumors, and their cells are like those in the originating tumor. For example, this means that if the breast cancer metastasizes to the lungs, the secondary tumor system consists of abnormal breast cells rather than abnormal lung cells. Thus, the tumor in the lungs is called metastatic breast cancer, not lung cancer. It is known from the metastatic procedures described herein and those known in the art that the sites of metastasis include, but are not limited to, lung, liver, brain, and bone. A "VEGF inhibitor" as used herein includes any substance that inhibits or binds to VEGF or VEGF-R. Unless otherwise indicated, all references to VEGF inhibitors used herein include those pharmaceutically acceptable salts, solvates, hydrates, and complexes thereof, and pharmaceuticals thereof. References to solvates, hydrates and complexes of the salts which are acceptable, including their polymorphs, stereoisomers and isotopically labeled forms. For example, a preferred VEGF inhibitor is sunitinib, which, as used herein, refers to sunitinib malate (ie, Sutent®) and any of the free bases of sunitinib and sunitinib. Other pharmaceutically acceptable salts. A preferred VEGF inhibitor is sunitinib (5_(5-fluoro-2-oxo-oxime, 2·dihydroindole_(3Z)_ylidene)), 4_ represented by the following formula Dimethyl_ih_pyrrole-3-carboxylic acid (2-diethylaminoethyl)-decylamine)··

It is a new oral cancer drug that shows efficacy in a variety of solid tumor types. Sunitinib targets multiple receptor tyrosine kinase inhibitors (including PDGFR, KIT, and VEGFR) as an effective and selective anti-angiogenic agent. Sunitinib or its L_malate is also known as smi248, SU011248, sunitinib malate (usan/wh〇 name) or sutentTM (l. malate). The term "sunitinib" as used herein includes 5-(5-fluoro-2-indolyl_u_dihydro. 弓bundo...subunitylmethyl)·2,4-di-fyl -1H-. Bilo-3-carboxylic acid (2-diethylaminoethyl)-decylamine and its pharmaceutically acceptable salts (including L-malate and sutentTM). </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; No. 2005-0059824 and J.Μ. Manley, MJ Kalman, BG Conway, CC Ball, JL Havens and R. Vaidyanathan, "Early Amidation Approach to 3-[(4-amido)pyrrol-2-yl]-2- Indolinones, J. Org. Chem. 68, 6447-6450 (2003). A preferred formulation of sunitinib and its L-malate is described in U.S. Patent Publication No. 2004-0229930 and PCT Publication No. WO 2004/024127. The preferred dosing regimen is described in U.S. Patent Publication No. 2005-01 82,122 and PCT Publication No. WO 2006/120557. The disclosures of these references are hereby incorporated by reference in their entirety in their entireties in the the the the the the the the the the For example, U.S. Patent Publication No. 2003-02-0264 No. 0 describes the combination of Compound 1 and a cyclooxygenase inhibitor. U.S. Patent Publication No. 2004-0152759 describes the combination of Compound 1 with several agents such as CPT-11 (irinotecan (CamptosarTM)) 'docetaxel and 5-fluoro Urine 0 bite (5-FU). Other references to sunitinib and its uses include U.S. Patent Nos. 7,211,600 and 7,119,209 and (: 1 'Publication No. 0 2006/101692, WO 2003/015608, WO 2003/035009, WO 2004/045523, and WO 2004/075775. The disclosures of these references are The manner in which the full text is incorporated is incorporated herein by reference. Other preferred VEGF inhibitors include, but are not limited to, 158393.doc • 18- 201217361 axitinib (6-[2-(methylamine-mercapto)) Phenylthio]dEPOpyridin-2-yl)vinyl]carbazole, AG-13676) (Pfizer Inc.): Η, ch3 〇%^Ν'

IT, which is a selective VEGF and a PDGF inhibitor, is described in U.S. Patent Nos. 6,534,524, 6,884,890 and 7, 141, 581, and PCT Publication No. WO 2008/122858 and WO 2006/123223, The disclosures of the literature are hereby incorporated by reference in their entirety to the extent of the entire disclosure of SU-14813 (5-[(Z)-(5-fluoro-2-oxo-l,2-dihydro-3H-)吲哚· 3-subunit) fluorenyl]-N-[(2S)-2-hydroxy-3-morpholin-4-ylpropyl]-2,4-dimethyl-1Η-° 3-曱醯amine) (Pfizer Inc.): 〇

It is a potent and selective oral inhibitor of the receptor tyrosine kinase (RTK), which is directly involved in a signaling cascade that triggers tumor growth, progression and survival. SU-14813 is a stimulator of VEGF, PDGFRa and PDGFRP, KIT and FLT3 158393.doc • 19-201217361, and is described in US Patent Nos. 6,653,308 and 7,247,627 and in references cited by sunitinib. The disclosures of these documents are hereby incorporated by reference in their entirety in their entirety in their entirety in the the the the the the the the Phenyl-4-yloxy)benzoxanthran-3-amine (PHzer Inc.):

It is a selective VEGF inhibitor, which is described in U.S. Patent No. 7,381,824 and PCT Publication No. WO 2007/017740, the disclosure of each of which is hereby incorporated by reference. Other VEGF inhibitors include, but are not limited to, Avastin (bevacizumab, Genentech), anti-VEGF monoclonal antibodies, and VEGF inhibitors, which are described, for example, in U.S. Patent Nos. 6,534,524, 5,834,504, 5, 883, 113, 5, 886, 020, 5, 792, 783, 6, 653, 308, and 6, 235, 764 (all references are hereby incorporated by reference in its entirety for all purposes) and WO 99/24440, WO 95/21613 , WO 99/61422, WO 98/50356, WO 99/10349, WO 97/32856, WO 97/22596, WO 98/54093, WO 98/02438, WO 99/16755 and WO 98/02437 (all such documents 158393.doc -20- 201217361 is incorporated herein by reference. The term "c_Met/HGFI^p formulation" as used herein includes any substance which inhibits or binds c-Met or its ligand HGFR. Unless otherwise indicated, all references to e-trapping T/HGFR inhibitors herein include, for pharmaceutically acceptable salts, solvates, hydrates and complexes, and pharmaceutically acceptable salts. References to solvates, hydrates, and complexes, including polymorphs, stereoisomers, and isotopically labeled forms thereof. A particularly preferred c-Met/HGFR inhibitor is klotinib ((R)-3-(H2,6_digas_3_fluorophenyl)ethoxy)_5_(1 ten) represented by the formula Bottom line _4_ base). Than bite _2_amine):

It is a new oral cancer drug that shows efficacy in a variety of solid tumor types. Klotinib targets the protein tyrosine kinase, including c-Met/HGFM ALK. Ke (4) Ni is also known as pF G234i () 66. As used herein, the term "kelotinib" includes (r)_Hi_(2,6-dichloro- 3 disordered phenyl)ethoxylated bite 4_yl>IHh4^)d(tetra)_2-amine and its medicine A salt that is acceptable for learning. 158393.doc -21 - 201217361 Klotinib, its synthetic and specific polymorphic systems are described in U.S. Patent No. 7,230,098, U.S. Patent Publication Nos. 2006-0046991 and 2008-0293769. The disclosures of these references are hereby incorporated by reference in their entirety. C-Met/HGFR inhibitors suitable for use in the practice of the present invention are described in, for example, WO 2007/0265272, WO 2009/068955, WO 2006/086484, WO 2005/030140, WO 2008/051808, U.S. Patent 4,923,986, U.S. Patent 7, All of these documents are incorporated herein by reference in their entirety. Another particularly preferred c-Met/HGFR inhibitor is PF-04217903 (2-(4-(3-(quinoline-6-ylindenyl)-3Η-[1,2,3) represented by the formula Triazolium [4,5-b]pyrazin-5-yl)-1Η-pyrazol-1-yl)ethanol):

It is a novel oral cancer drug. Klotinib targets the protein tyrosine kinases, including c-Met/HGFR and ALK. The term "PF-0421 7903" as used herein includes (R)-3-(1-(2,6-dioxa-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-) -1 - pyrazol-4-yl)pyridin-2-amine and pharmaceutically acceptable salts thereof, including its phosphate PF-04217903, its synthetic and specific polymorphic systems are described in U.S. Patent No. 7,732,604 No. 12/745,411 (corresponding to 158 393. doc -22-201217361 PCT Publication No. WO 2009/068955), the disclosure of each of which is hereby incorporated by reference. Other c-Met/HGFR inhibitors suitable for use in the practice of the invention include, but are not limited to, AMEP (Bioalliance), EMD-1204831 (Merck KgaA/EMD Serono) 'INCB-028060 (Incyte/Novartis) 'ARQ197 (ArQule) ' AMG102 (Amgen) and RG-3638 (Roche/Genentech), and such inhibitors are described in WO 2007/0265272, WO 2009/068955, WO 2006/086484 &gt; W02005/030140 'WO 2008/051808, US Patent 4,923,986, U.S. Patent No. 7,713,969, U.S. Patent No. 7,579,473, the entireties of each of The term "pharmaceutically acceptable salts" includes acid addition salts and base salts (including di-salts). Suitable acid addition salts are formed from acids which form non-toxic salts. Examples include acetate, aspartic acid. Salt, benzoate, besylate, bicarbonate/carbonate, hydrogen sulfate/sulfate, borate, camphorsulfonate, citrate, ethanedisulfonate, ethanesulfonate, A Acid salt, fumarate, glucoheptonate, gluconate, glucuronate, hexafluorophosphate 'sea benzoate, hydrochloride/chloride, hydrobromide/telluride , hydroiodide/iodide, isethionate, lactate, malate, maleate, malonate, methanesulfonate, sulfonate, naphthate, 2-naphthalene Sulfonate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate/hydrogen phosphate/dihydrogen phosphate, gluconate, stearic acid Salts, succinates, tartrates, benzenesulfonates, and trifluoroacetates. Suitable base salts are formed by the formation of non-toxic salts. Examples include aluminum salts, Arginine, benzathine penicillin 158393.doc -23- 201217361 salt, calcium salt, choline salt, diethylamine salt, glycolamine salt, glycinate, ammonium salt, magnesium salt, glucosamine Salt, alcohol amine salt, potassium salt, sodium salt, tromethamine salt and zinc salt. For a review of suitable pharmaceutically acceptable salts, see Stahl and Wermuth, "Handbook of Pharmaceutical Salts.

Properties,Selection,and Use"(Wiley-VCH, Weinheim

Germany, 2002), the disclosure of which is hereby incorporated by reference in its entirety, in its entirety, the pharmaceutically acceptable salt of the compound of the present invention can be readily prepared by mixing the compound solution with the desired acid or base, where appropriate. . The salt may be precipitated from the solution and collected by filtration or may be recovered by evaporation of the solvent. The degree of ionization in the salt can change from fully ionized to almost non-ionized. The compounds of the invention may exist in unsolvated as well as solvated forms. The term "solvate" is used herein to describe a molecular complex comprising a compound of the invention and one or more pharmaceutically acceptable solvent molecules (e.g., ethanol). When the solvent is water, the term "hydrate" is used. The pharmaceutically acceptable solvates of the present invention include hydrates and solvates wherein the solvent for crystallization can be isotopically substituted (e.g., 〇2〇, dr acetone, d DMSO) 〇6-position, such as 2h, respectively. 3h The present invention also encompasses compounds labeled with isotopes which, in addition to or having a plurality of atoms having atomic substitutions other than the atomic mass or mass atomic mass or mass number normally found in nature, are inhibited by VEGF as described herein. The agent h_Met inhibitor is the same. Examples of isotopes which may be incorporated into the compounds of the invention include gas, charcoal, nitrogen, oxygen, dish, sulfur, gas and gas. 14 Ό C, 15N, 158393.doc -24· 201217361 p p, s, 18f and 36ci. The compound of the present invention containing the above isotope and/or the other isotope of the other atom and the pharmaceutically acceptable salt of the compound belong to the present invention. Certain labeled isotopic compounds of the invention, for example, and the likes of radioisotopes t such as 4 and 4c, are suitable for drug and/or matrix tissue distribution assays. Deuterated isotope (ie 3H) and carbon 14 isotope (ie 14c) are particularly preferred for their ease of preparation and detectability. In addition, they are replaced by heavier isotopes of H (ie 2H) due to greater metabolism. Stability may provide certain therapeutic advantages, such as increased in vivo half-life or reduced dosage requirements, and thus may be preferred in some circumstances. Compounds labeled with an isotope can generally be prepared by substituting a reagent described for the unlabeled compound with a readily available labeled isotope reagent for the non-isotopically labeled reagent. Also included within the scope of the invention are complexes such as scorpions, drug-host inclusion complexes in which the drug and the host are present in stoichiometric or non-stoichiometric amounts in comparison to the solvates mentioned above. φ includes a pharmaceutical complex containing two or more organic and/or inorganic components which may be in stoichiometric or non-stoichiometric amounts. The resulting complex can be ionized, partially separated or not ionized. For a review of such complexes, see Haleblian^J Pharm Sci, 64 (8), 1269-1288 (1975 #-8^), the disclosure of which is incorporated herein in its entirety by reference. DETAILED DESCRIPTION Unless otherwise indicated, all references herein to c-met/hgfr inhibitors or VEGF inhibitors include pharmaceutically acceptable salts, solvates, hydrates and complexes thereof, and pharmaceutically acceptable thereto. References to solvates, hydrates and complexes of the salts of I58393.doc -25- 201217361, including polymorphs, stereoisomers and isotopically labeled forms thereof, are acceptable. In the study of the present invention, we tested the efficacy of sunitinib (RTKI targeting VEGF, PDGFR-β, CSF-1R 'c-Kit and Fit3) in a preclinical model and identified resistance/sensitivity to therapy. The tumor. Analysis of protein lysates in resistant or sensitive tumors revealed more HGF in the former. The combination treatment with a highly selective c-Met inhibitor (PF-04217903 and Klotinib (PF-02341066)) and sunitinib has an additive effect in inhibiting tumor growth compared to either single agent. These results indicate the functional role of the HGF/c-Met axis in sunitinib resistant tumors. We studied the pattern of resistance to sunitinib. Sunitinib was clinically approved for metastatic RCC (renal cell carcinoma) and imatinib-resistant GIST (gastrointestinal stromal tumor) and in several The test is used for a variety of tumor types (including lung cancer, colon cancer and breast cancer) (Smith JK et al, Emerging roles of targeted small molecule protein-tyrosine kinase inhibitors in cancer therapy. Oncology research 2004*14 (4- 5): 175- 225 ; van der Veldt AA et al, Sunitinib for treatment of advanced renal cell cancer: primary tumor response. Clin

Cancer Res 2008; 14(8): 2431-6). The efficacy of sunitinib in murine cell lines in the current study and comparison with recent reports (Shojaei F et al, Nature biotechnology 2007; Shojaei F et al, PNAS 2009) indicate blocking receptors in inhibiting tumor growth The signaled signaling is as effective as blocking the ligand. Future studies may determine whether there are unique signs of blocking the VEGF pathway in comparison to inhibitory ligands in preclinical models. 158393.doc •26· 201217361 The reason why we focus our research on the c-Met pathway is that several studies have shown that this pathway plays an important role in development, tumor formation, and angiogenesis. There are at least three different tumor orders. Mechanisms involved in the activation of the c_Met pathway: i) ligand-mediated activation via binding to HGF; ii) amplification of the c-Met locus; and 丨丨丨^在卜]^"receptor kinase domain In the role of dogs (Smith JK et al, Oncology research 2004; van der Veldt AA et al, Clin Cancer Res 2008). It is worth noting that 'because no cell line shows different concentrations in vitro or in vivo The c-Met inhibitor has a significant response, so our data indicate that the c-Met system is activated by binding to HGF in the study of the present invention. Therefore, in our model, a large concentration of HGF is required to activate c_Met in the tumor. Pathway. The lack of HGF expression in tumor or endothelial cells indicates that the matrix acts as a source of HGF in resistant tumors. In vitro data indicate that although both tumor and endothelial cells are exposed to HGF', only the latter can proliferate. Thus, the hypothesis that angiogenesis is the main target of HGF in these tumors is provided. Although the concentration in the tumor is large, HGF does not appear to be released in the clearing. The evaluation of the spinal cord population further confirms the direct effect of HGF on angiogenesis. Because c-Met inhibition and HGF up-regulation in sensitive tumors do not affect the kinetics of spinal cord cells in peripheral blood or tumors. In order to test whether sunitinib treatment affects metastasis in preclinical models, we in situ To study the metastatic potential of H460-GFP and C〇1〇205-GFP tumors. GFP transduction allowed tumor growth in the viscera of live animals. Five people supervised and performed all procedures in Anti-Cancer Inc, Anti- Cancer

Inc. is a leading company in the field of tumor biology and preclinical model imaging. I58393.doc •27- 201217361 — ο Therefore, these studies have shown unexpected results, ie C-Met inhibitors can potentially serve as any One of the treatment options for tumor types is partially added to VEGF inhibitors, where sutent treatment triggers activation of the c-Met pathway via the following mechanisms: i) up-regulation of HGF and/or ii) overexpression of c-Met receptor And/or iii) c-Met receptor activation. The cancers to which the present invention can be used for a specific application include, but are not limited to, colon cancer, non-small cell lung cancer, melanoma, renal cancer, hepatocellular carcinoma, glioblastoma multiforme, and pancreatic neuroendocrine tumor. Administration Compounds suitable for use in the present invention can be administered in any manner that provides benefits to the patient. Different combinations of c-Met/HGFR inhibitors and VEGF inhibitors may require changes in the route of administration and dosage regimen. Different cancer types may also require changes in the route of administration and dosage regimen. The present invention encompasses all changes in administration based on any combination of factors, such as the type of cancer being treated, the c-Met/HGFR inhibitor administered, the VEGF inhibitor administered, and the administration thereof. A preferred dosing regimen for c-Met/HGFR inhibitors, a preferred dosing regimen for the VEGF inhibitor administered, or the needs of the patient being treated. Compounds suitable for use in the present invention can be administered by continuous administration. Continuous administration can include administration of a VEGF inhibitor followed by administration of a c-Met/HGFR inhibitor. Continuous administration can include administration of a c-Met/HGFR inhibitor followed by administration of a VEGF inhibitor. Continuous administration can be based on a preferred dosing regimen based on a c-Met/HGFR inhibitor or a VEGF inhibitor. Continuous administration can be readily adjusted based on any one or more of the following factors: type of cancer treated 158393.doc -28-201217361, c_Met/HGFR inhibitor administered, formulation administered v, administered Preferred administration of c_Met/HGFR inhibitor: preferred dosing regimen for administration of VE_formation or need of patient/, a compound suitable for use in the present invention can be c-Met by reversing the inhibitor /HGFR inhibitors are administered in conjunction with administration. Concomitant administration can be in the form of a single pharmaceutical composition comprising at least one c_Met/HGFR inhibitor in combination with at least one VEGF inhibitor, or at least one c-Met/HGFR inhibitor and at least one VEGF inhibitor. Administration is carried out by administration of different administration routes, pharmaceutical compositions or different administration vehicles. Whether administered by a mono-f pharmaceutical composition, continuous administration or concomitant administration: two or more compounds suitable for the present invention or metabolites thereof may be simultaneously present in a patient. Further, the compound or its metabolite which is suitable for use in the present invention may be absent from the patient, whether by a single pharmaceutical composition, continuous administration or concomitant administration (4). Routes of administration may include, but are not limited to, any of the following: oral administration, parenteral administration, topical administration, inhalation/intranasal administration, rectal/vaginal administration, administration via the eye, or familiarity with the technique. Any other known route of administration &amp; The route of administration of the c_Met/HGFR inhibitor and VEGF inhibitor suitable for use in the present invention may be the same or different. Oral Administration Compounds suitable for use in the present invention can be administered orally. Oral administration can include sputum in order for the compound to enter the gastrointestinal tract, or buccal or sublingual administration can be used to thereby directly enter the bloodstream from the mouth. Formulations suitable for oral administration include solid formulations, such as lozenges, capsules containing 158393.doc -29-201217361 microparticles, liquids or powders, buccal ingots (including fillings containing liquids), chewables, Multiparticulates and Nanoparticles, Gels, Solid Solutions, Liposomes, Bismuth (including Mucoadhesives), Oval Suppositories, Sprays, and Liquid Formulations 0 Liquid formulations include suspensions, solutions, syrups and elixirs. The formulations may be used as a filler in soft or hard capsules' and generally include a pharmaceutically acceptable carrier (eg water, ethanol, polyethylene glycol, propylene glycol, decyl cellulose or a suitable oil) And one or more emulsifiers and/or suspending agents. Liquid formulations can also be prepared by reconstituting, for example, solids from a sachet. The compounds of the present invention are also useful in rapidly dissolving, rapidly disintegrating dosage forms such as those described in the Expert Opinion in Therapeutic Patents of Liang and Chen, 11 (6), 981-986 (2001), the disclosure of which is incorporated herein by reference. This article is incorporated herein by reference. For lozenge dosage forms, the dosage may comprise from i% by weight to 80% by weight of the dosage form, more typically from 5% to 6% by weight of the dosage form, depending on the dosage. In addition to the drug, the tablet usually also contains a disintegrant. Examples of the disintegrant include sodium starch glycolate, sodium carboxymethyl cellulose, calcium carboxymethyl cellulose, croscarmellose sodium, crosslinked polyvinylpyrrolidone, polyvinylpyrrolidone, methyl fiber. , microcrystalline cellulose, hydroxypropyl cellulose substituted by low carbon yard, temple powder, pregelatinized starch and sodium alginate. In general, the disintegrant will constitute from 1% to 25% by weight of the dosage form, preferably from 5% to 2% by weight. Binders are generally used to impart cohesive properties to the formulation of the tablet. Suitable binders include microcrystalline cellulose, gelatin, sugar, polyethylene glycol, natural and synthetic rubber, 158393.doc -30- 201217361 polyvinylpyrrolidone, pregelatinized starch, hydroxypropyl cellulose and hydroxypropyl Methyl cellulose. Tablets may also contain diluents such as lactose (monohydrate, spray dried monohydrate, anhydrate, and the like), mannitol, xylitol, dextrose, sucrose, sorbitol, microcrystals Cellulose, starch and dihydrogen phosphate dihydrate. Tablets may also optionally include surfactants (such as sodium lauryl sulfate and polysorbate 80) and slip agents (such as ceria and talc). When present, the amount of surfactant is typically from 2% to 5% by weight of the tablet, and the slip agent is typically from 0.2% to 1% by weight of the tablet. Tablets also generally contain a lubricant such as magnesium stearate, calcium stearate, stearic acid, sodium stearyl fumarate, and a mixture of magnesium stearate and sodium lauryl sulfate. The lubricant is usually from 25% by weight to 3% by weight, preferably from 0.5% by weight to 3% by weight of the tablet. /. The amount exists. Other S-known ingredients include antioxidants, colorants, flavoring agents, preservatives, and taste masking agents. Exemplary lozenges contain up to about 80% by weight of the drug, about 10% by weight. /〇 to about 90% by weight of the binder, from about 5% by weight to about 85% by weight of the diluent, from about 2 parts by weight to about 1 weight. /. The disintegrant and about 〇 25 weight ° /. To about 1% by weight of lubricant. The tablet blend can be compressed directly or by a roller to form a tablet. Alternatively, the tablet blend or blend portion may be wet granulated, dry granulated or melt granulated, melt condensed or extruded prior to tableting. The final formulation may include one or more layers and may be coated or uncoated; or encapsulated. Formulations of lozenges are described in detail in H. Lieberman and L. Lachman 158393.doc • 31 - 201217361, "Pharmaceutical Dosage Forms: Tablets, Volume 1," Marcel Dekker, NY, NY, 1980 (ISBN 0-8247- The disclosure of this document is incorporated herein by reference in its entirety. Solid formulations for oral administration can be formulated to provide immediate and/or improved release formulations. Modified release formulations include delayed release formulations, sustained release formulations, pulsed release formulations, controlled release formulations, targeted release formulations, and stylized release formulations. Suitable modified release formulations are described in U.S. Patent No. 6,1,6,864. Details of other suitable release techniques such as high energy dispersions and osmotic and coated granules can be found in Verma et al, Pharmaceutical Technology On-line, 25(2), i-14 (2001). The use of chewing ingots to achieve controlled release is described in W〇 00/35298. The disclosures of these references are hereby incorporated by reference in their entirety. Parenteral administration No needle injection Carbonation But for the direct administration of the compounds of the invention to blood, muscle or internal organs: Suitable methods for parenteral administration include intravenous administration, intra-arterial administration, intraperitoneal administration, intralesional administration, intraventricular administration, intraurethral administration, intrathoracic administration, intra-medication, intramuscular administration, and subcutaneous administration. Suitable devices for parenteral administration include needle (including microneedle) injectors and infusion techniques. Parenteral formulations are typically aqueous solutions which may contain a formulation such as a complex and a buffer (preferably buffered to a positive value of 3 to 9) but for some applications, it may be more suitably formulated as Sterile non-aqueous solution or form to be used in conjunction with, for example, sterile, pyrogen-free water (iv) (iv) 158393.doc -32-201217361. For example, preparation of parenteral formulations under sterile conditions by lyophilization can be readily accomplished using standard pharmaceutical techniques well known to those skilled in the art. The solubility of the compounds of the invention for preparing parenteral solutions can be enhanced by the use of suitable formulation techniques, such as incorporation of dissolution enhancers. Formulations for parenteral administration can be formulated as immediate and/or improved release formulations. Modified release formulations include delayed release formulations, sustained release formulations, pulsed release formulations, controlled release formulations, targeted release formulations, and stylized release formulations. Thus, the compounds of the present invention can be formulated as solid, semi-solid or shake-reducing liquids for administration as a phytopharmaceutical reservoir providing improved release of the active compound. Examples of such formulations include drug-coated vascular stents and? (31^ eight microspheres. Topical administration may also be carried out by topical administration of a compound of the invention to the skin or mucous membranes or transdermal administration. Typical formulations for this purpose include gels, hydrogels, lotions , solutions, creams, ointments, powders, dressings, foams, films, skin patches, bridges, implants, latitude, fibers, end bands and microemulsions. Typical carriers include alcohols, water, mineral oil, liquid stone soil, white rock, glycerin, polyethylene glycol, and propylene glycol. Infiltration enhancers can be incorporated, see, for example, Finnin and Μ% (10)^

Pharm Sci, 88 (1〇) 955-QSS; 958 (1st month of 1999). Other methods of local administration include delivery by electroporation, iontophoresis, ultrasonic drug penetration therapy, iontophoresis, and microneedle or needle-free (eg, p〇wderjectTM, called (10)TM, etc.) injections. . The disclosures of these references are incorporated herein by reference in their entirety by reference. Formulations for topical administration can be formulated as immediate and/or improved release formulations. Modified release formulations include delayed release formulations, sustained release formulations, pulsed release formulations, controlled release formulations, dry release formulations, and stylized release formulations. Inhalation/intranasal administration The compounds of the invention may also be inhaled in dry powder form, usually in the form of a dry powder (alone, as a mixture 'for example, dry blended with lactose; or as a mixed component granule, for example mixed with a phospholipid such as phospholipid choline) Or as an aerosol spray with or without the use of a suitable propellant (such as hydrazine, hydrazine, hydrazine, 2_tetrafluoroethane or 1,1,1,2,3,3,3-heptafluoropropane) A pressure vessel, a pump, a nebulizer, an atomizer (preferably an atomization or a nebulizer that utilizes electrohydrodynamics to produce a fine mist) is administered intranasally or by inhalation. For intranasal use, t's powder may include bioadhesives, such as polyglucosamine or cyclodextrin. M. Barbill water....... 虱Vatomizer) 匕4匕 (nebuhzer) contains this A solution or suspension of the inventive compound, which comprises, for example, ethanol, an aqueous solution of ethanol or a dispersion, solubilization or elongation of the active substance*, a replacement brake, a propellant as a solvent, and optionally a surfactant, such as a detachment agent.尺山 4 sugar alcohol trioleate, oleic acid or oligomeric lactic acid. Prior to use in dry powder or suspension quenching, the drug product is micronized to a size suitable for insufflation (usually less than 5 microns). This can be achieved by any suitable comminution method, such as spiral jet milling, fluidized bed jet milling, 158393.doc 201217361 grinding, supercritical fluid processing to form nanoparticles, high pressure homogenization or spray drying. Capsules for use in an inhaler or insufflator (for example, made of gelatin or HPMC), foaming agents and cartridges may be formulated to contain a compound of the invention, a suitable powder base such as lactose or starch, and A powder mixture of potency modifiers such as /- leucine, mannitol or magnesium stearate. Lactose may be anhydrous or in the form of a monohydrate, preferably the latter. Other suitable excipients include dextran, glucose, maltose, sorbitol, xylitol, fructose, sucrose, and trehalose. Suitable solution formulations for nebulizers that use electrohydrodynamics to produce a fine mist may contain from 1 Kg to 20 mg of the compound of the invention per actuation, and the actuation volume may be from 丨μ1 to 丨〇〇μ1 Variety. Typical formulations include the compounds of the invention, propylene glycol, sterile water, ethanol, and sodium chloride. Alternative solvents for alternative propylene glycol include glycerin and polyethylene glycol. Suitable perfumes (such as menthol and evomentol Oevomentho) or sweeteners such as saccharin or sodium saccharin may be added to such formulations of the invention for inhaled/intranasal administration. Formulations for inhaled/intranasal administration can be formulated for immediate and/or improved release formulations using, for example, poly(DL-lactic acid-ethanol I) (PGLA). Modified release formulations include delayed release formulations, sustained release formulations, pulsed release formulations, controlled release formulations' targeted release formulations, and stylized release formulations. In the case of dry powder inhalers and aerosols, the dosage unit is determined by means of a metered amount of wide gate. A unit according to the present invention is typically formulated to administer 158393.doc -35.201217361 with a metered dose or "puff" containing the desired amount of the compound of the invention. The total dose per dose can be administered in a single dose, or more usually in divided doses within one day. Rectal/Vaginal Administration The compounds of the invention may be administered rectally or vaginally, for example, in the form of a suppository, pessary or enemas. Cocoa butter is a typical suppository base, but various alternatives can be used as appropriate. Formulations for rectal/vaginal administration may be formulated as immediate and/or modified release formulations. Modified release formulations include delayed release formulations, sustained release formulations, pulsed release formulations, controlled release formulations, dry release formulations, and stylized release formulations. Ophthalmic administration The compounds of the present invention may also be administered directly to the eye or ear in the form of micronized suspensions or drops of solution in isotonic, pH adjusted sterile physiological saline. Other formulations suitable for ocular and otic administration include ointments, biodegradable (eg, absorbable gel sponges, collagen), non-biodegradable (eg, polyoxygenated) implants, wafers, Contact lenses and microparticles or vesicular systems such as vesicles or liposomes. The t compound (such as cross-linked polyacrylic acid, polyethylene glycol, vitreous acid, cellulose polymer (such as hydroxypropyl decyl cellulose, hydroxyethyl cellulose or sulfhydryl cellulose) or heteropolysaccharide The polymer (eg, gellan gUm) is incorporated with a preservative such as benzylammonium sulfate. These formulations can also be delivered by iontophoresis. Formulations for trans- or per-ear administration can be formulated for immediate and/or improved release formulation 158393.doc -36· 201217361. Modified release formulations include delayed release formulations, sustained release formulations, pulsed release formulations, controlled release formulations, light release formulations or stylized release formulations. Other Techniques In order to be used in any of the above modes of administration, the compounds of the present invention may be combined with soluble macromolecular entities such as cyclodextrins and suitable derivatives thereof or polymers containing polyethylene glycol to improve their solubility, Dissolution rate, taste masking, bioavailability and/or stability. For example, the discovery of a drug-cyclodextrin complex is generally applicable to most dosage forms and routes of administration. Both inclusion complexes and non-inclusion complexes can be used. As an alternative to direct mismatch with the drug, the cyclodextrin can be used as an auxiliary additive, i.e., as a carrier, diluent or solubilizer. The most commonly used for these purposes are alpha-cyclodextrin, beta-cyclodextrin and gamma-cyclodextrin, examples of which can be found in

The disclosures of these references are hereby incorporated by reference in their entirety in their entireties in the the the the the the the the the the the the the the the the the Dosage The amount of active compound administered will depend on the subject being treated, the severity of the condition or condition, the rate of administration, the nature of the compound, and the judgment of the designated physician. However, the effective dose is usually in the range of from about 0.001 mg to about _ beer per kg body weight per day in a single or divided dose, preferably from about 1 mg/kg/day to about 35 mg/kg/ day. For a human of 7 kilograms, the effective dose will be from about 0'07 mg to about 7 mg per day, preferably from about 0.7 mg to about 2500 mg per day. In some cases, agents below the lower limit of the above I58393.doc •37-201217361 range are not sufficient to cause any harmful side effects, while in other cases, the amount is usually more such that it can be used even more The kit 1 of each part of the large agent is divided into several small doses for reading throughout the day. "If the specific compound or condition is treated with the fine compound of the active compound, it is desirable to administer σ, so it is desirable to describe the two or more pharmaceutical compositions, &gt; Suitable for the combination of a combination of ', ', ', ', and σ. Therefore, the present invention: 'and includes two or more separate pharmaceutical compositions (at least one of which has 3 Compounds of the invention) and components for the independent storage of such compositions, such as cereals, separate bottles or divided collar bags. Examples of such kits are well known for use in the preparation of capsules, capsules and the like. The present invention is particularly suitable for use in, for example, oral and parenteral administration of different dosage forms, for the application of separate compositions at different dosage intervals, or to separate compositions for mutual drip. To aid compliance, the kit usually includes a dosing instructions' and may have a memory aid. Example General Procedures Tumor Implantation and Treatment Nude mice are purchased from Jackson (ME, USA) or Charles Rivers Laboratories (MA, USA) and According to Pfizer IACUC (institutional animals) Feeding by the guidelines provided by the Institutional Animal Care and Use Committee. All tumor cell lines in the current study were obtained from ATCC (American Tissue Culture Center; Manassas, VA) and cultured in supplemented with amylinamide (2 mM) and fetal bovine serum (FBS; 10%) in RPMI 1640 158393.doc -38- 201217361 (invitrogen, CA). At the time of implantation, tumor cells (1 x 10 6 cells per mouse) were resuspended In 100 μΐ medium and 100 μΐ matrigel growth factor reduced (BD BioSciences, CA), and subcutaneously implanted in one of the flanks. Once daily, oral administration was used at 80 mg. /kg sunitinib or PF-04217903 (45 mg/kg) or a combination of two compounds to treat tumor-bearing mice. Tumor volume is used as (Zou HY et al, An orally available small-molecule inhibitor of c -Met, PF-2341066, exhibits cytoreductive antitumor efficacy through antiproliferative and antiangiogenic mechanisms. Cancer research 2007; 67(9): 4408-17) The caliper measurement method described is evaluated. HUVEC (human umbilical vein endothelial cells) and C166 cells were purchased from Lonza Inc. (Lonza, CA) and ATCC, respectively. In an in vitro assay, HUVECs were grown in EBM2 medium supplemented with a growth factor mixture supplied by a supplier (Lonza Inc, Lonza CA), and C166 was grown in DMEM supplemented with FBS (10%) (invitrogen) , CA). General Procedure 2: Histology The formalin-fixed tumor was embedded in paraffin and sectioned using a cryostat (4 μιη). Tumor sections were stained with mouse anti-CD3 1 antibody (BD BioSciences, CA) using DAB immunostaining protocol in the laboratory. To quantify the vascular surface area (VSA), tumor images at 2 〇 x magnification were obtained from each slide and each treatment. Next, the region of CD31+ cells was highlighted and the pixels were quantified using Image-Pro software (MediaCybernetics, Bethesda, Maryland). Vascular density is obtained by dividing CD3 1-pixel by 158393.doc -39- 201217361

Images of all regions of the pixel to calculate β General Procedure 3: ELISA The tumor samples were snap frozen in liquid nitrogen and kept at _7〇〇c. Total protein was extracted from each sample and quantified using BCA protein assay (Pierce, IL). The content of HGF and total Met in tumors and serum is measured by the ELISA kit (R&amp;d System, CA) using the protocol provided by the manufacturer. A similar method is applied to the measurement of conditioned medium or tumor lysate. HGF content. General Procedure 4: Flow Cytometry

Single cell suspensions were obtained from tumors by mechanical disruption followed by lysis of red blood cells using RBC lysis buffer (eB ioscience, CA). Peripheral blood mononuclear cells (PBMNC) were also obtained by lysing red blood cells using RBC lysis buffer (eBioscience, CA). In; 4. Tumor cells and PBMNC were stained with anti-mouse CDUb_APC (BD Biosciences, CA) and anti-mouse Grl-PE (BD Biosciences, CA) for 30 minutes, followed by washing with PBS-FBS (3〇/〇). Times. Analysis of stained samples in a FACS Calibur instrument (BE) Bioscience, CA) using ceu Quest (BD)

Biosciences, CA) or FCS Express (De Novo Inc, CA) software analysis of the incidence of CD11 b+Grl + cells in tumors and jk fluids. Similarly, measuring c-Met expression on the cell surface will grow in vitro. Cell lines and endothelial cells to bind? Anti-mouse 0-^161; (68丨〇3(^611(^,08) staining. Exclude dead cells from the analysis'. The cells were analyzed with 7-AAD (7-amine) on a Calibur instrument. Base-actinomycin D; BD Biosciences, CA) staining. General procedure 5: in vitro assay 158393.doc -40- 201217361 will include B16F1, Tib6, EL4, LLC and endothelial cell lines, HUVEC and C166 per well 104 cells were seeded in tissue culture-treated 24-well plates. Cells were grown in standard medium as described above. Sunitinib, PF-04217903 at different concentrations (2 μηι, 0.2 μηι, and 0.02 μηη) Cells were treated for 4 days in combination with the two compounds. The efficacy of the compounds was measured by counting cells in a Coulter Counter (Coulter Counter Machine, BD Biosciences, CA). A similar method was used, using three different concentrations (10 ng/ml; 100 ng/ml; and 200 ng/ml) of each ligand to assess the role of HGF or VEGF in cell proliferation. Example 1: Identification of cell lines resistant or sensitive to sunitinib treatment. Nitinib resistance

Murine cell lines including B16F1, Tib6, EL4 and LLC were implanted into nude mice (n=6) using General Procedure 1. Cells (1 x 106 per mouse) were seeded in 100 μΐ medium supplemented with 100 μΐ low growth factor base gum. After the implantation, the sputum was started with sunitinib (80 mg/kg) or vehicle. The data represents one of two independent studies. Similar to recent reports using anti-VEGF monoclonal antibodies (Shojaei F et al, Tumor refractoriness to anti-VEGF treatment is mediated by CD1 lb+Grl+ myeloid cells. Nature biotechnology 2007; 25(8): 911-20), B16F1 and Tib6 Tumors were sensitive to sunitinib and EL4 and LLC tumors were found to be resistant to the therapy. As illustrated in Figure 1, B16F1 (Figure 1A) and Tib6 (Figure 1B) tumors were sensitive to sunitinib, while EL4 (Figure 1C) and LLC (Figure 1D) tumors showed treatment for treatment shortly after the start of therapy. Resistance. These results are previously reported with the efficacy of the anti- 158393.doc -41 - 201217361 VEGF mAb in the same cell line (Sh〇jaei F et al.

Human, Nature biotechnology 2007; Shojaei F et al, PNAS 2009), this indicates a similar trend in tumor reactivity in the comparison of anti-VEGF and VEGF pathway small molecule inhibitors. Comparison of sunitinib and axitinib resistance Using general procedure 1, EL4 or LLC tumors were implanted into nude mice (n=10) as described. Treatment with vehicle, sunitinib (8 〇 mg/kg SID) or axitinib (35 mg/kg BID) was started one day after the implantation. The tumor growth inhibition of 'EL4 or LLC' is similar between sunitinib and axitinib. Thus, inhibition of the PDGF-R, CSF-1R and C-Kit pathways does not provide any advantage in tumor growth inhibition compared to inhibition of VEGF alone. In addition to the VEGF pathway, sunitinib also targets several other signaling pathways, such as PDGF-RB (platelet-derived growth factor receptor), CSF_ 1R (community stimulating factor #c_kit (5_[5 discovered by Sun L et al. _ Said_2_Sideoxy-1,2-dihydroindole-(3Z)-ylidene]yl-2,4-dimethyl-1H_pyrrole_3_decanoic acid (2-diethylamine) Base ethyl) guanamine, a novel tyrosine kinase inhibitor targeting vascular endothelium and platelet-derived growth factor receptor tyrosine kinase,

Journal of Med Chem 2003;46(7):1116-9). In order to understand whether the inhibition of sunitinib on these other pathways plays a role in the response to therapy, we compared sunitinib with axitinib (a selective VEGF inhibitor (Hu-) in resistant tumors. Lowe DD et al, N〇nclinical antiangiogenesis and antitumor activities of axitinib (AG-013736), an oral, potent, and selective inhibitor of vascular endothelial growth factor receptor tyrosine kinases 1, 2, 3. 158393.doc •42- 201217361

Clin Cancer Res 2008; 14(22): 7272-83)) (Figure 1E and Figure IF). Since no tumor responded to treatment, sunitinib and axitinib showed no statistically discernible effects on tumor growth in EL4 (Figure 1E) and LLC (Figure 1F). Therefore, these tumors appear to be resistant to other targets of sunitinib, such as PDGF-RB, CSF-1R and c-kit. Example 2: HGF behaves differently in resistant tumors and mainly targets endothelial cells in serum and tumors in tumors treated with sunitinib and in tumors using general procedure 1, B16F1, Tib6, EL4 and LLC cells were seeded in nude mice (n=6) and treated with sunitinib or vehicle as described. At the end of the analysis, total protein was extracted from each sample and quantified using General Procedure 3. ELISA data showed that the serum HGF concentration gradually decreased from mice without tumors to all tumor-bearing mice, indicating that HGF is highly concentrated in the tumor and is not released into the blood circulation (Fig. 2a). &lt;&apos;&apos;&apos;&apos;s resistant tumors have significantly higher HGF levels in tumors than in sensitive tumors, especially in sunitinib-treated mice (p&lt;〇5) (Fig. 2B). These data indicate that larger concentrations of HGF may be associated with resistance to sunitinib. Identification of the source of HGF in tumors. Resistant and sensitive cell lines as well as endothelial cells (HUvec &amp; ci66) and sputum (positive control for HGF expression) were inoculated into tissue culture at 5 χ 〇 5 cells per well. Dispose of each hole of the 6-well plate. The cells were incubated for 72 hours under normoxia (2% 〇2) and anoxic conditions (1% (6) incubator t. Conditioned medium was removed from each well and cells were harvested and stored in thieves. Use 158393.doc •43· 201217361 General procedure 3 performed HGF ELISA. NGF was not enriched in tumors and endothelial cells regardless of oxygen concentration, indicating that the stromal compartment may be the main source of HGF in the tumor mass (Fig. 2C) and human fibroblast strain (Mrc5). (Jiang WG et al., Reduction of stromal fibroblast-induced mammary tumor growth, by retroviral ribozyme transgenes to hepatocyte growth factor/scatter factor and its receptor, c-MET. Clin Cancer Res 2003;9(11):4274-81)- Therefore, NIH3T3 cells were highly enriched in HGF in both protein lysates and conditioned media. HGF mainly used endothelial cells to study the flow of c-Met on the cell surface of tumor cells and endothelial cells (HUVEC and C166) according to the general procedure 4'. Cellular measurement. Flow cytometric analysis indicated that tumor cells exhibited minimal amounts of c_Met on the cell surface, while c-Met showed more performance in endothelial cells (p&lt;0 〇5) (Fig. 2D) Example 3: HGF involves tumor/endothelial cell proliferation According to the general procedure 5, tumor cells (B16F1, Tib6, EL4 and LLC) and endothelial cells (HUVEC and C166) were seeded at a density of 104 per well. Tissue cultured 24-well plates were grown in common medium supplemented with 1% FBS. Cells were treated with three different concentrations (10 ng/ml; 1 ng/ml and 200 ng/ml) of HGF and VEGF. After 4 days of incubation, cells were counted using a Coulter counter. Both VEGF and HGF did not affect cell proliferation in tumor cells (Fig. 3 A). Conversely, both growth factors promoted proliferation of endothelial cells (Fig. 3B). : Combination therapy in sensitive or resistant tumors (sunitinib plus pF_158393.doc • 44 - 201217361 04217903) Efficacy according to the general procedure 1 'B16F1, Tib6, EL4 and LLC cells (1 X per mouse) 106 cells were implanted into nude mice (n=6). Treatment was started one day after implantation, and tumor volume was measured twice a week. As illustrated in Figures 4A and 4B, PF-04217903 monotherapy was performed. Does not inhibit tumor growth in any sensitive tumor, indicating tumors in these Lack of activation c_ Met path. Alternatively, the lack of PF-04217903 monotherapy response due to the presence of VEGF may also be indicative of the role of the VEGF pathway in tumor growth. In addition, there was no significant difference in tumor growth between combination therapy in B16F1 (Fig. 4A) and Tib6 (Fig. 4B) sensitive tumors compared with sunitinib monotherapy. This indicates a lack of c_Met pathway in these tumors. . However, 'combination therapy significantly inhibited the growth of two resistant tumors (EL4'. Fig. 4C' and LLC; Fig. 4D) compared to either single regimen (ρ&lt;〇·〇5). Example 5: Quantification of blood vessels in resistant or sensitive tumors The vascular structures of both sensitive and resistant tumors were analyzed according to General Procedure 2 (Fig. 5A). The CD3 1 stained image is selected from each part (four images in each part and a total of twenty-four images), and the vascular density is calculated by dividing the signal intensity of the CD31 + pixels by the total area of the image. . In sensitive tumors, sunitinib significantly reduced vascular density (p &lt; 〇 〇 5) as measured by cD31 positive areas compared to vehicle treated tumors. However, compared with single-agent and combination regimens, sunitinib did not affect the degree of gold tube. Inhibition of e_Met by PF-G4217903 did not significantly alter the sensitivity or anti-tumor of the IT structure. The progress of VEGF in promoting tumor growth and angiogenesis was observed. In resistant tumors, 'sunitinib inhibits the vascular system compared to vehicle-treated tumors. In addition, in resistant tumors, combination therapy significantly reduces vascular surface area compared to sunitinib alone. Endothelial cell selectivity According to the general procedure 5, tumor cells (B16F1, Tib6, EL4 and LLC; Fig. 5B) and endothelial cells (HUVEC and C166; Fig. 5C) were seeded in a 12-well tissue culture plate at a density of 104 per well. And grown in a standard medium as described. The cells were incubated in a cell incubator for 4 days with sunitinib, PF-0421 7903 or a combination of the two compounds at three different concentrations (including 2 μηη, 〇2 μηι and 〇〇2 added to each well). Counted in a Coulter counter as described. Sunitinib and the combination significantly inhibited endothelial cell proliferation at pharmacologically relevant concentrations (Fig. 5B), whereas tumor cells were neither affected by the agent nor affected by the combination (Fig. 5C). Example 6: Combination of sunitinib and PF_02341〇66 inhibits in situ tumor metastasis in H46〇nsclc (non-small cell lung cancer) Cells are implanted in situ in the lungs of nude mice. Briefly, tumor lines The tumor was collected from the subcutaneous implant at the log phase by subcutaneously implanting H460-GFP cells (lxl〇6 cells per mouse) into nude mice and maintained in RPMI-1640 medium (Sigma_Alrdrich). The necrotic tumor was removed from the medium and the living tissue was cut into 2 〇1112 to 25 cm2 slices. The recipient mice were anesthetized with isoflurane and the surgical area was disinfected with iodine and alcohol. Chest wall, followed by the left lung And the intercostal incision between the fourth ribs. For each mouse, two identical H460-GFP tumor fragments were implanted into the lung surface. The thoracic cavity was closed after implantation and was made by thoracentesis 158393.doc •46- 201217361 Lung re-expansion. All surgical procedures were performed in a HEPA-filtered laminar flow shield using a magnifying microscope (01yinpus). The mice were allowed to recover from the surgical intervention for 72 hours before the start of treatment. Group (n=l 0), starting treatment with vehicle, sunitinib, PF-02341066 and combination (combination of sunitinib and pf-02341066). The mice were administered 23 days after implantation. Euthanasia, and tumors were collected for histochemical analysis. On the last day of the study, open green fluorescent probe imaging (GFP images) was used to obtain animal images to assess primary tumor growth and to other organs. Transfer. As illustrated in Figure 6, the GFP image (Figure 6A) and the final tumor weight (Figure 6B) do not reveal a significant difference between the vehicle and any single agent or combination therapy. Or a combination regimen does not completely inhibit primary tumor growth, suggesting that angiogenic mediators other than VEGF and HGF may be involved in H460 tumor growth. Analysis of metastases in the lung or lymph nodes indicates that sunitinib does not inhibit tumor metastasis However, PF-02341066 or combination therapy reduced the occurrence of metastasis (Fig. 6C). Example 7: Combination of sunitinib and PF_〇2341066 inhibited the metastasis of c〇LO205 colorectal tumors in situ tumors C〇1〇2 〇5-GFP cells (5×丨〇6 cells per mouse) were subcutaneously implanted into nude mice to produce tumor strains. Mice were euthanized when tumor growth reached log phase. Tumor slabs were isolated and maintained in RPMI 164 〇 medium (Sigma-Aldrich) to remove necrotic tumors and maintain living tissue for intra-embryo implantation. The recipient's surgical area was disinfected with iodine and alcohol, and the mice were anesthetized with a mixture of Ketamine, Acepromazine and xylazine 158393.doc •47·201217361 (Xylazine). In the cecal implant, the left middle abdomen of the nude mouse was cut open to about 1 cm and the colonic serosa was removed to allow implantation of two tumors (2 cm2 to 2.5 cm2 each). In order to close the abdominal cavity, the abdominal muscles and skin are sutured separately using silk sutures. All procedures were performed under a dissecting microscope in a laminar flow shield. Mice that were allowed to be implanted in situ at the austhesia recovered consciousness for 72 hours from the surgical intervention and started treatment when the tumor reached 75 mm3 to 1 〇〇 mm3. The mice were divided into 4 groups and administered with vehicle, sunitinib, and ? 17_〇2341〇66 and a therapeutic compound of sunitinib in combination with PF-02341066. GFP images of the treated animals were obtained weekly and the study was terminated 4 weeks after tumor implantation. Tumor size was measured by calculating the vertical minor size (W, width) and major size (L, length) using a FluorVivo imaging system (indec Bi〇systems). Analysis of tumor-bearing mouse GFP images indicated no significant difference between sunitinib, vehicle and PF-02341066 (Fig. 7A). However, the combinedly treated mice showed a significant decrease or decrease in tumor volume and tumor weight (Fig. 7B). In addition, analysis of metastasis to lymph nodes and other areas in the colon showed that both sunitinib and pF_02341066 did not inhibit metastasis, whereas combination therapy completely blocked metastasis since no mice showed metastatic lesions to other organs (Fig. 7C). . [Simple description of the map]

Figure 1 - Identification of cell lines resistant or sensitive to sunitinib treatment. B16F1 (Fig. 1Α) and Tib6 (Fig. 1Β) tumors were sensitive to sunitinib and found that EL4 (Fig. 1C) and LLC (Fig. 1D) tumors were resistant to the therapy. (Fig. 1 and Fig. 1F) inhibition of PDGF-R, CSF-1R and c-Kit pathways did not provide any advantage in tumor growth inhibition compared to inhibition of VEGF alone 158393.doc-48-201217361. Sunitinib is completely similar to acitretin in tumor growth inhibition in EL4 (Fig. 1E) or LLC (Fig. 1F). Figure 2 - Endothelial cells (rather than tumor cells) are primarily targeted by the HGF/c_Met axis. HGF measurements in serum (Figure 2A) and tumor (Figure 2B) in vehicle treated and sunitinib-treated tumors. Bars represent the mean concentration of HGF in tumors treated with sunitinib or vehicle. (Fig. 2C) HGF is mainly expressed in the matrix compartment in the tumor mass. (Fig. 2D) c_Met is mainly expressed in endothelial cells. The image is a representative histogram comparing the e_Met performance (red column) with the isotype control (black column) for each column. When c_Met is rarely expressed in tumor cells, it is highly enriched in endothelial cells. Figure 3. Proliferation of endothelial cells (rather than tumor cells) is induced by hgjt. Tumor cells (B16F1, Tib6, EL4, LLC; Fig. 3A) and endothelial cells (HUVEC and C166; Fig. 3B) were seeded in tissue culture treated 24-well plates at a density of 1〇4 per well, and grown in Supplemented with 10/〇FBS in normal medium. Cells were treated with three different concentrations (10 ng/ml; 100 ng/ml &amp; 2 〇 () ng/ml) of HGF and VEGF. After 4 days of incubation, cells were counted using a c〇u® counter. Asterisks indicate significant differences in HGF-treated or VEGF-treated cells compared to PBS. Figure 4: The combination of sunitinib and PF-04217903 has an additive effect compared to sunitinib monotherapy. The efficacy of combination therapy (sunitinib plus PF-04217903) in sensitive or resistant tumors. B16F1 (Fig. 4A), Tib6 (Fig. 4B), EL4 (Fig. 4C), and LLC (Fig. 4D) cells (lxi〇6 cells per mouse) were implanted into nude mice (n=6). Treatment was initiated one day after implantation and 156393.doc •49·201217361 The tumor volume was measured twice a week. Each figure represents an SEM. The asterisk indicates when the difference is different (p&lt;〇.〇5). Represents the mean tumor volume, and the difference between the nitnitibin and the combination therapy. Figure 5. The inhibition of angiogenesis is one of the mechanisms by which combination therapy affects tumor growth. Quantification of blood vessels in rot in resistant or sensitive tumors (Fig. 5A). The image of CD31 staining &amp; is selected from each part (each Qiu knife (Valley P divided into four images and a total of twenty-four images), and the vascular density system 鹑', 曰 by CD31 + pixel signal The intensity is divided by the total area of the image. Bridge you I _ τ _ know' The uneven blood vessel density iSEM. The asterisk indicates significant when comparing sunitinib 盥 Newman', '··&' Differences (p &lt; 〇〇 5) (Fig. 5B and Fig. 5C) 〇 Endothelial cells (rather than tumor cells) were sensitive to the combination treatment with sunitinib and PF 04217903. The data represent the comparison of two independent research stars. There was a significant difference between sunitinib alone and vehicle, and ♦ a significant difference in comparing the effect of combination therapy with the number of cells of sunitinib or 04217903 at the corresponding concentration. Figure 6: Single agent and combination therapy The effectiveness of the H460 lung orthotopic tumor in the growth of primary and metastatic tumors. GFP image (Fig. 6A) and final tumor weight (Fig. 6B) were not in vehicle and any single agent. Or a significant difference between the combination treatments. Analysis of the metastasis in the bark (Fig. 6C) showed that sunitinib did not inhibit tumor metastasis, whereas klotinib or combination therapy reduced metastasis. Figure 7: Single agent and combination therapy in C〇1〇205 colon in situ The effectiveness of tumors in the growth of primary and metastatic tumors. The GFP image of tumor-bearing mice (Fig. 7A) indicates no significant difference between sunitinib, vehicle and klotinib, but indicates combination The difference between treatment and sunitinib and klotinib was 158393.doc -50- 201217361. The mice treated by combination showed a significant decrease or decrease in primary tumor volume and primary tumor weight (Fig. 7B). Analysis of lymph nodes and other regional metastases in the colon (Fig. 7C) showed that neither sunitinib nor PF-02341066 inhibited metastasis, whereas combination therapy completely blocked metastasis. 158393.doc -51 -

Claims (1)

201217361 VII. Scope of Application: 1. The use of a VEGF inhibitor and a C-Met/HGFR inhibitor for the preparation of a medicament for treating cancer in a mammal in need thereof, wherein the medicament comprises a therapeutically effective amount The VEGF inhibitor and the therapeutically effective amount of the c-Met/HGFR inhibitor. * 2. The use of claim 1, wherein the mammal is a human. 3. The use of claim 1, wherein the cancer is selected from the group consisting of colon cancer, non-small cell lung cancer, melanoma, renal cancer, hepatocellular carcinoma, glioblastoma multiforme, and pancreatic neuroendocrine Tumor. The use according to any one of claims 1 to 3, wherein the VEGF inhibitor is selected from the group consisting of sunitinib, SU-14813, PF-337210, bevacizumab (bevacizumab) ) and axitinib (axitinib). 5. The use of any one of claims 1 to 3, wherein the VEGF inhibitor is sunitinib. 6. The use of any one of claims 1 to 3, wherein the VEGF inhibitor is axitinib. The use of any one of claims 1 to 3, wherein the VEGF inhibitor is *bevacizumab. The use of any one of claims 1 to 3, wherein the VEGF inhibitor is SU-14813. The use of any one of claims 1 to 3, wherein the VEGF inhibitor is PF-337210. The use according to any one of claims 1 to 3, wherein the C-Met/HGFR inhibition 158393.doc 201217361 agent is crizotinib or PF-04217903. 11. 12. 13. 14. 15. 16. The use of any of claims 1 to 3, wherein the 〇 _ _ / / 11 (} 1 ^ inhibitor is klotinib. If the request items 1 to 3 The use of any of the VEgf inhibitors is sunitinib and the c_Met/HGFR inhibitor is klotinib. The use of any one of claims 1 to 3 wherein the VEGF inhibitor is Axi The C-Met/HGFR inhibitor is ketotinib. The use of any one of claims 1 to 3 wherein the VegF inhibitor is bevacizumab and the c-Met/HGFR inhibitor is Klebsi The use of any one of claims 1 to 3, wherein the VEGF inhibitor is SU_14813 and the c-Met/HGFR inhibitor is klotinib. The use of any one of claims 1 to 3, Wherein the VEGF inhibitor is PF-33721 0 and the c-Met/HGFR inhibitor is klotinib. 158393.doc