JP2022046903A - Therapeutic agent for diffuse gastric cancer - Google Patents

Abstract

To provide molecular targeted drugs effective on diffuse gastric cancer.SOLUTION: Provided is a therapeutic agent for treating diffuse gastric cancer comprising a dual inhibitor of MET and VEGF receptor 2. The invention also provides a therapeutic agent for treating diffuse gastric cancer comprising a VEGF receptor 2 inhibitor for use in a combination therapy with MET inhibitor, and a therapeutic agent for treating diffuse gastric cancer comprising a MET inhibitor for use in a combination therapy with a VEGF receptor 2 inhibitor. Further, the invention provides a method for treating diffuse gastric cancer in a patient by inhibiting MET and VEGF receptor signaling pathway activity, comprising administering one or more agents for inhibiting the activity of the MET and VEGF receptor 2 signaling pathway.SELECTED DRAWING: None

Description

The present invention relates to a therapeutic agent for diffuse gastric cancer, which comprises a double inhibitor of MET and VEGF receptor 2. The present invention includes a therapeutic agent for diffuse gastric cancer containing a VEGF receptor 2 inhibitor for use in combination therapy with a MET inhibitor, and a MET inhibitor for use in combination therapy with a VEGF receptor 2 inhibitor. Regarding therapeutic agents for diffuse gastric cancer. Furthermore, the present invention is a method for treating diffuse gastric cancer in a patient by suppressing the activity of the MET and VEGF receptor signaling pathways, and is one for suppressing the activity of the MET and VEGF receptor signaling pathways. The present invention relates to a method comprising administering the above-mentioned agent to the above-mentioned patient.

Diffuse gastric cancer consists of poorly differentiated adenocarcinoma, signet ring cell carcinoma, etc., and tends to progress faster and have a poorer prognosis than non-diffuse gastric cancer consisting of well-differentiated adenocarcinoma. In particular, scirrhous gastric cancer included in diffuse gastric cancer is characterized by having severe interstitial fibrosis, diffuse infiltration of cancer cells, and often undifferentiated adenocarcinoma cells (Non-Patent Document 1). Scirrhous gastric cancer frequently develops peritoneal carcinomatosis (cancerous peritonitis) consisting of peritoneal dissemination of cancer cells and a large amount of ascites, resulting in an extremely poor prognosis (Non-Patent Documents 1 and 2).

Due to the characteristic of diffuse gastric cancer, which is rapidly growing, most advanced cases are diagnosed in the state of stage IV with cancerous ascites, so treatment by radical surgical resection can hardly be expected (Non-Patent Document 2, Non-Patent Document 2, Non-Patent Document 3). As standard treatment for unresectable advanced recurrent gastric cancer, SP therapy using a combination of S-1 (tegafur / gimeracil / oteracil potassium combination drug) and cisplatin (CDDP) is performed (Non-Patent Document 4), but it is not. The therapeutic effect in diffuse gastric cancer is lower than that in diffuse gastric cancer.

On the other hand, with the advent of anti-HER2 therapy, personalized medicine centered on anti-HER2 therapeutic agents is also drawing attention in the treatment of gastric cancer. Regarding HER2-positive gastric cancer, although the survival-improving effect of trastuzumab was clarified in an international joint study (ToGA), the proportion of HER2-positive gastric cancer in the total gastric cancer is not high (about 10 to 20%). In addition, HER2 is not observed in diffuse gastric cancer. Therefore, for gastric cancer other than HER2-positive gastric cancer, a search for a more useful new molecular-targeted drug that contributes to personalized medicine is being conducted.

Gastric cancer is divided into "early gastric cancer" and "advanced gastric cancer" according to the degree of invasion. 50-60% of advanced gastric cancer in which the cancer invades deeper than the muscularis propria is diffuse gastric cancer, of which around 20% It is said to be scirrhous stomach cancer. However, no effective molecular-targeted drug for advanced gastric cancer is known. For example, an international collaborative study (AVAGAST) was conducted to verify the additional effect of bevacizumab on chemotherapy for advanced and recurrent gastric cancer, but its effectiveness was not confirmed. In addition, the efficacy of panitumumab or cetuximab on chemotherapy for advanced gastric cancer was not confirmed in the study (REAL3 study, EXPAND study).

Treatment methods based on an understanding of the cancer progression mechanism characteristic of diffuse gastric cancer have been studied (Patent Documents 1 and 2). Patent Document 1 discloses a therapeutic agent for diffuse gastric cancer containing a MEK inhibitor and a combination therapy of an mTOR inhibitor and a MEK inhibitor for diffuse gastric cancer. Patent Document 2 describes a pharmaceutical composition for treating diffuse gastric cancer in a patient containing an EGF receptor inhibitor having ErbB1 inhibitory activity and ErbB4 inhibitory activity, as well as the above-mentioned EGF receptor inhibitor and anti-VEGF receptor 2 antibody. And / or a combination therapy with a cMET inhibitor is disclosed.

WO2019 / 230595A1 WO2020 / 100969A1

Zhao L. et al., Cancer Science, December 2013, vol.104, No12, pp.1640-1646. Yasumoto K. et al., Clin Cancer Res 2011; 17: 3619-3630. Yasumoto K. et al., Cancer Res 2006; 66: (4) 2181-2187. Koizumi W. et al., Lancet Oncol. 9 (3): 215-21 (2008).

As mentioned above, treatment methods for diffuse gastric cancer are being developed. To study multiple pathways that contribute to cell proliferation and cancer progression mechanisms, and to develop treatment methods that bring about higher therapeutic effects. Is required. Therefore, it is an object of the present invention to provide a new therapeutic method for diffuse gastric cancer and a therapeutic agent used therein.

As a result of intensive studies to solve the above problems, the present inventor has found that double inhibition of MET and VEGF receptor 2 has a beneficial effect in the treatment of diffuse gastric cancer. The present invention is based on this finding.

According to the present invention, the following inventions are provided.
[1] A therapeutic agent for diffuse gastric cancer, which comprises a double inhibitor of MET and VEGF receptor 2.
[2] Double inhibitors of MET and VEGF receptor 2 are cabozantinib (BMS-907351), cabozantinib malate (XL184), gorbatinib (E7050), MGCD-265, BMS-794833 and Ningentiib (CT-053). , DE-120, CT053PTSA), the therapeutic agent according to [1].
[3] A therapeutic agent for diffuse gastric cancer containing a VEGF receptor 2 inhibitor for use in combination therapy with a MET inhibitor.
[4] A therapeutic agent for diffuse gastric cancer containing a MET inhibitor for use in combination therapy with a VEGF receptor 2 inhibitor.
[5] The therapeutic agent according to [3] or [4], wherein the MET inhibitor and the VEGF receptor 2 inhibitor are co-administered or sequentially administered in combination therapy.
[6] The MET inhibitor is sabolitinib (AZD6094, HMPL-504), crizotinib (PF-02341066), foretinib, PHA-665752, SU11274, SGX-523, BMS-777607, JNJ-38877605, tivantinib (ARQ19). Group consisting of PF-04217903, capmatinib (INCB28060), BMS-754807, AMG-208, MK-2461, AMG-458, NVP-BVU972, norcantharidin, AMG337, S49076, NPS-1034 and Therapinib (LY2801653). The therapeutic agent according to any one of [3] to [5], preferably sabolitinib (AZD6094, HMPL-504).
[7] The therapeutic agent according to any one of claims 3 to 6, wherein the VEGF receptor 2 inhibitor is an anti-VEGFR2 antibody, preferably ramucirumab.
[8] The therapeutic agent according to any one of [1] to [7], wherein the diffuse gastric cancer is scirrhous gastric cancer.
[9] The therapeutic agent according to any one of [1] to [8], wherein the diffuse gastric cancer has cancerous peritonitis.
[10] The therapeutic agent according to any one of [1] to [9], wherein the diffuse gastric cancer is unresectable or has metastasis.
[11] A method for treating diffuse gastric cancer in patients by suppressing the activity of MET and VEGF receptor signaling pathways.
A method comprising administering to the patient one or more agents for suppressing the activity of the MET and VEGF receptor signaling pathways.
[12] Containing administration of a dual inhibitor of MET and VEGF receptor 2 to the patient, or administration of a combination therapy of MET inhibitor and VEGF receptor 2 inhibitor to the patient [11]. The method described.

FIG. 1 shows survival curves for the seven treatment and control groups tested in vivo. It is a comparison of the combination therapy "HER1 / 4-TKI + anti-VEGFR2 antibody" and "MET-TKI + anti-VEGFR2 antibody". The vertical axis is the survival rate, and the horizontal axis shows the number of days. The survival curve was prepared by the Kaplan-Meier method and a log rank test was performed. ^* P <0.05 is a comparison with the control group. FIG. 2 shows survival curves for the seven treatment and control groups tested in vivo. Comparison of multiple inhibitors (cabozantinib, gorbatinib) and combination therapy "MET inhibitor + VEGFR inhibitor". The vertical axis is the survival rate, and the horizontal axis shows the number of days. The survival curve was prepared by the Kaplan-Meier method and a log rank test was performed. ^* P <0.05 is a comparison with the control group.

The description of the present invention described below may be based on typical embodiments and specific examples, but the present invention is not limited to such embodiments. In this specification, the numerical range represented by using "-" means a range including the numerical values before and after "-" as the lower limit value and the upper limit value.

Diffuse gastric cancer is one of the most malignant intractable gastric cancers. Gastric cancer is classified as type 1 (ulcer type), type 2 (ulcer localized type), type 3 (ulcer infiltration type) to type 4 (diffuse infiltration type) according to the classification method (Ballman classification) when the morphology is viewed with the naked eye. ). In some cases, the type 5 cannot be classified into types 1 to 4. As used herein, "diffuse gastric cancer" refers to type 3 (ulcer infiltration type) and type 4 (diffuse infiltration type), and scirrhous gastric cancer refers to type 4 (diffuse infiltration type). Unless otherwise specified in the present specification, "diffuse gastric cancer" includes scirrhous gastric cancer. The stage of gastric cancer will be described later.

MET (also referred to as c-MET, cMet) is a transmembrane receptor tyrosine kinase also known as hepatocyte growth factor receptor (HGFR). MET includes gastric cancer, lung cancer, colon cancer, breast cancer, bladder cancer, head and neck cancer, ovarian cancer, prostate cancer, thyroid cancer, pancreatic cancer, and cancer of the central nervous system (CNS). Often amplified, mutated, or overexpressed in many types of cancer. MET is produced as a single-stranded precursor, which is cleaved at the furin cleavage site into α and β subunits, both linked by disulfide bonds (Nat Rev Mol Cell Biol 4, 915). -925 (2003)). Activation of MET by hepatocyte growth factor (HGF), a ligand for MET, is an excessive cellular process including proliferation, motility, invasiveness, metastasis, epithelial-mesenchymal transition, angiogenesis / wound healing, and tissue regeneration. (Christensen et al., Cancer Lett 225: 1-26, 2005; Peters and Adjei, Nat Rev Clin Oncol 9: 314-26, 2012).

MET amplification is a molecular abnormality that occurs in gastric cancer, and MET signaling is essential for the survival of MET amplification-positive gastric cancer cells, but MET amplification is rare (2%) in advanced gastric cancer (non-patent literature). 1). On the other hand, HGF is overexpressed in most gastric cancers, suggesting the involvement of HGF in the progression of gastric cancer. Malignant ascites in patients with cancerous peritonitis contains high levels of HGF, and paraclinic HGF-induced activation of MET-mediated signaling pathways plays an important role in the pathogenesis of cancerous peritonitis in scirrhous gastric cancer. It has been reported to fulfill (Non-Patent Document 1).

The MET inhibitor showed a high antitumor effect in a mouse cancerous peritonitis model using MET gene-amplified gastric cancer and scirrhous gastric cancer cells (Non-Patent Document 1). However, several clinical trials with TKI (tyrosine kinase inhibitor) and antibodies targeting the MET receptor for the entire gastric cancer have been conducted, and sufficient clinical usefulness has been confirmed in all of the trials. Not. Here, MET gene-amplified gastric cancer, which accounts for 2% of all gastric cancers, may be difficult to be classified as diffuse gastric cancer macroscopically, but has the characteristics of diffuse gastric cancer from the viewpoint of the molecular mechanism of progressive development. (Nakajima M et al., Cancer 1999; 85: 1894-1902).

VEGF receptor 2 (also referred to herein as VEGFR2) means vascular epidermal growth factor receptor (VEGFR) 2. VEGFR is a transmembrane receptor tyrosine kinase. It is characterized by an extracellular region with seven immunoglobulin-like regions and an intracellular tyrosine kinase region. So far, VEGFR2 (KDR / Flk1), as well as VEGFR1 (Flt1) and VEGFR3 (Flt4) have been reported. VEGFR2 is expressed on vascular endothelial cells and plays a central role in angiogenesis, angiogenesis and pathogenic angiogenesis. In its involvement in malignant tumors, the binding of VEGF-A to VEGFR2 is Ras / Raf / MAPK. Leads to endothelial cell proliferation via activation of the / ERK and PI3K / Akt / mTOR pathways (Nat Rev Clin Oncol 2009, 6 (9): 507-518). Anti-VEGF receptor 2 antibody lambsilumab given weekly paclitaxel In combination with the method, it is recommended for secondary chemotherapy for unresectable advanced / recurrent gastric cancer (gastric cancer treatment guidelines for doctors, revised January 2018, 5th edition).

The present invention provides a method of treating diffuse gastric cancer in a patient by suppressing the activity of MET and VEGF receptor signaling pathways and therapeutic agents used therein.

The present inventor has found that in the search for a therapeutic method for diffuse gastric cancer, double inhibition of the signal transduction pathways of MET and VEGF receptor 2 has a beneficial effect on the treatment of diffuse gastric cancer. .. In the examples described below, inhibition of the signal transduction pathway of MET and VEGF receptor 2 is EGF receptor and VEGF receptor 2 using an EGF receptor inhibitor (HER1 / 4-TKI) and an anti-VEGF receptor 2 antibody. It was revealed that it has a higher therapeutic effect than the inhibition of the signal transduction pathway of. In addition, double inhibition of MET and VEGF receptor 2 (combination therapy and double inhibitor) was compared to treatment with MET inhibitor alone or VEGF receptor 2 inhibitor alone in mouse treatment experiments. , It showed a very excellent effect of suppressing ascites production and prolonging the prognosis, while side effects such as decreased activity and weight loss were hardly observed. It was known that MET and VEGF receptor 2 are deeply involved in cancer growth and progression, respectively, but by blocking the signaling pathways of MET and VEGF receptor 2, diffuse gastric cancer It is surprising that it has a beneficial effect on treatment. As mentioned above, MET amplification is rare (2%) in advanced gastric cancer, and several clinical trials with TKI (tyrosine kinase inhibitor) and antibodies targeting the MET receptor in the entire gastric cancer have been conducted. However, sufficient clinical usefulness has not been confirmed in any of the studies. Furthermore, Non-Patent Document 2 reports that EGFR and its ligands, particularly amphiregulin and HB-EGF, are important for the development of peritoneal carcinomatosis (cancerous peritonitis) in gastric cancer patients. Since the therapeutic effect of EGF receptor inhibitors on patients with diffuse gastric cancer is expected, it inhibits the signal transduction pathways of MET and VEGF receptor 2 rather than the signal transduction pathways of EGF receptor and VEGF receptor 2. It was unexpected that a higher therapeutic effect would be obtained.

As used herein, the therapeutic effect is cure or remission of the disease being treated, and / or alleviation or amelioration of one or more of the symptoms associated with the disease, even though the patient may still be suffering from the disease. Means. In addition to the effects of reducing and ameliorating disease-related symptoms, it also has the effect of slowing the rate of exacerbations of such symptoms. Therapeutic effect also includes stopping or delaying the progression of the disease, whether or not improvement in symptoms is achieved.

Ｔ／Ｎ／ＭおよびＳｔａｇｅの定義は、『胃癌取扱い規約第１５版』（『ＵＩＣＣ－ＴＮＭ分類第８版』）による。 The stage of gastric cancer is comprehensively judged by the degree of cancer invasion and the presence or absence of lymph node / distant metastasis. The TNM classification integrates the evaluation of three factors: the degree of cancer invasion (T factor), the presence or absence of lymph node metastasis and its number (N factor), and metastasis to distant organs (distant metastasis, M factor). (Table 1, Source: Gastric Cancer Treatment Guidelines for Doctors, Revised January 2018, 5th Edition).

The definitions of T / N / M and Stage are based on the "Stomach Cancer Handling Regulations 15th Edition"("UICC-TNM Classification 8th Edition").

The stage of diffuse gastric cancer to which the treatment method of the present invention is targeted is not particularly limited, and for example, stages I, IIA, IIB, III, IVB and IVA can be treated. Stage I is early gastric cancer and stages II, III and IV are advanced gastric cancer. The preferred treatment target is advanced diffuse gastric cancer. In particular, the method of the present invention provides an excellent therapeutic method for advanced diffuseness of stage IV. The therapeutic method of the present invention is also effective for diffuse gastric cancer having cancerous peritonitis (distant metastasis). Gastric cancer with cancerous peritonitis (distant metastasis) is an unresectable, advanced, recurrent gastric cancer that is classified as stage IV. In addition, the therapeutic method of the present invention is effective for both primary gastric cancer and metastatic gastric cancer.

(Therapeutic agent 1)
The present invention provides a therapeutic agent for diffuse gastric cancer, comprising a double inhibitor of MET and VEGF receptor 2.

Double inhibitors of MET and VEGF receptor 2 generally reduce or prevent the activity of MET and VEGF receptor 2 simultaneously or doubly. Appropriate MET and VEGF receptor 2 dual inhibitors can be identified by assaying for inhibitory effects on MET and VEGF receptor 2 expression and / or activity. Inhibitors reduce expression or activity by at least a detectable amount in an appropriate assay. Inhibitors can, for example, reduce expression or activity by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or at least 90%. Any suitable dual inhibitor of MET and VEGF receptor 2 may have tyrosine kinase inhibitory activity and / or inhibit signal transduction. The double inhibitor of MET and VEGF receptor 2 may be a multiple inhibitor having other inhibitory activity in addition to MET and VEGF receptor 2.

In one embodiment, the dual inhibitors of MET and VEGF receptor 2 are cabozantinib (BMS-907351), gorbatinib (E7050), MGCD-265, BMS-794833 and Ningettiib (CT-053, DE-120, CT053PTSA). It is selected from the group consisting of. The double inhibitor can include pharmaceutically acceptable salts of these listed, for example, for cabozantinib, cabozantinib malate (XL184).

Cabozantinib (BMS-907351) is N- (4- (6,7-dimethoxyquinoline-4-yloxy) phenyl) -N'-(4-fluorophenyl) cyclopropane-1,1-dicarboxamide (CAS Registry Number). : 849217-68-1). Cabozantinib malate (XL184) is N- {4-[(6,7-dimethoxyquinoline-4-yl) oxy] phenyl} -N'-(4-fluorophenyl) cyclopropane-1,1- Dicarboxamide mono- (2S) -malate (CAS Registry Number: 1140909-48-3). Golbatinib (E7050) is 1,1-cyclopropanedicarboxamide, N- [2-fluoro-4-[[2-[[[4- (4-methyl-1-piperazinyl) -1-piperidinyl] carbonyl] amino]. ] -4-Pyridinyl] Oxy] Phenyl] -N'-(4-Fluorophenyl)-(CAS Registry Number: 928037-13-2). MGCD-265 is N-[[[3-fluoro-4-[[2- (1-methyl-1H-imidazol-4-yl) thieno [3,2-b] pyridin-7-yl] oxy] phenyl. ] Amino] thioxomethyl] -benzeneacetamide (CAS Registry Number: 875337-44-3). BMS-794833 is N- (4- (2-amino-3-chloropyridine-4-yloxy) -3-fluorophenyl) -5- (4-fluorophenyl) -4-oxo-1,4-dihydropyridine- 3-Carboxamide (CAS Registry Number: 1174046-72-0). Ningettiib (CT-053, DE-120, CT053PTSA) is 1H-pyrazole-4-carboxamide, N- [3-fluoro-4-[[7- (2-hydroxy-2-methylpropoxy) -4-quinolinyl]]. Oxy] phenyl] -2,3-dihydro-1,5-dimethyl-3-oxo-2-phenyl- (CAS Registry Number: 1394820-69-9).

Cabozantinib, exemplified as a double inhibitor of MET and VEGF receptor 2, is a small molecule compound having an inhibitory effect on kinases such as VEGF receptor 2, MET, AXL (Cancer Res, 2011, 71 (14), 4758. -4768). Cabozantinibrinate is approved in the United States for the treatment of patients with advanced metastatic medullary thyroid cancer (MTC), and in Japan for patients with unresectable or metastatic renal cell carcinoma. Approved as a therapeutic agent.

Golbatinib (E7050), exemplified as a double inhibitor of MET and VEGF receptor 2, is a tyrosine kinase inhibitor of VEGF receptor 2 and MET, and treatment with gorbatinib has been shown to prolong the survival of cancer-bearing mice. (Cancer Sci, 2010, 101 (1), 210-215).

MGCD-265, exemplified as a dual inhibitor of MET and VEGF receptor 2, is a tyrosine kinase inhibitor of VEGF receptor 2 and MET (Bioorg Med Chem Lett. 2008 May 1; 18 (9): 2793-8). ). BMS-794833, exemplified as a dual inhibitor of MET and VEGF receptor 2, is a tyrosine kinase inhibitor of VEGF receptor 2 and MET (WO2009 / 094417). Ningentiib (CT-053, DE-120, CT053PTSA), exemplified as a double inhibitor of MET and VEGF receptor 2, is a small molecule compound having an inhibitory effect on kinases such as VEGF receptor 2, MET, AXL (CT-053, DE-120, CT053PTSA). Cancer Res 2014; 74 (19 Suppl): Abstract nr 1755. Doi: 10.1158 / 1538-7445. AM2014-1755).

The therapeutic agent of the present invention can be administered according to known clinical achievements. The double inhibitor of MET and VEGF receptor 2 may be administered to an adult (body weight 60 kg) in an amount of 100 μg to 10 g, 500 μg to 10 g, or 1 mg to 1 g per day. When the double inhibitor of MET and VEGF receptor 2 of the present invention is cabozantinib malate, the dose as cabozantinib is 20-60 mg per day, preferably 40-60 mg per day, and more. It is preferably 60 mg per day.

Whether or not the above-mentioned double inhibitor of MET and VEGF receptor 2 (or the MET inhibitor or VEGF receptor 2 inhibitor described later) has an anticancer effect on diffuse gastric cancer is determined by in vitro cell growth inhibition. It can be confirmed by a test or a therapeutic effect evaluation test in a diffuse gastric cancer model. In an in vitro cell proliferation inhibition test, for example, a gastric cancer cell line is cultured with a double inhibitor of MET and VEGF receptor 2 (or a MET inhibitor or VEGF receptor 2 inhibitor described later) to grow cancer cells. Can be assessed to see if it is inhibited. In addition, a gastric cancer cell line cultured with cytokines and growth factors to proliferate cancer cells is further combined with a double inhibitor of MET and VEGF receptor 2 (or a MET inhibitor or VEGF receptor 2 inhibitor described later). It can be cultured to assess whether the growth of cancer cells is inhibited. As the cytokines and growth factors, for example, types that have been confirmed to be present in high concentrations in cancerous ascites of patients with diffuse gastric cancer as compared with non-cancerous ascites can be used. HB-EGF (heparin binding-epidermal growth factor-like growth), an amphiregulin with a concentration 2 to 5 times higher than that of non-cancerous abdominal water, is found in cancerous abdominal water associated with diffuse gastric cancer patients. It has been reported that factor), HGF (hepatocyte growth factor) and the like are present (Non-Patent Documents 1 and 2).

In the therapeutic effect evaluation test in a diffuse gastric cancer model, a double inhibitor of MET and VEGF receptor 2 (or a MET inhibitor or VEGF receptor 2 inhibitor described later) was applied to animals such as mice transplanted with gastric cancer cells. It can be carried out by administration and evaluation of the therapeutic effect. Evaluation can be performed by dissecting euthanized mice and observing the number and size of abdominal tumors, the amount of ascites, and the condition of nodules in the peritoneum. Assessments may be performed on a regular basis with general condition observations as well as body weight and food intake measurements. In addition, ascites retention can be evaluated by observing the appearance of the abdomen, palpation, and the like. The therapeutic effect may be evaluated by administering a double inhibitor of MET and VEGF receptor 2 (or a MET inhibitor or VEGF receptor 2 inhibitor described later) at the same time as transplantation for about 5 days. After confirming the growth of the tumor, a double inhibitor of MET and VEGF receptor 2 (or a MET inhibitor or VEGF receptor 2 inhibitor described later) may be administered and evaluated. As the model animal, rodents such as rats, primates such as rabbits, mini pigs, and cynomolgus monkeys can be used in addition to mice. In addition to the xenograft model using human cancer cells, an allogeneic transplantation model, an orthotopic transplantation model, a chemically carcinogenic model, and a genetically modified animal model can be used. When using the cancer cell transplantation model, it may be ectopic transplantation or ectopic transplantation.

Examples of gastric cancer cell lines include NUGC4, OCUM-1, MKN45, MKN7, MKN28, TMK-1, NKPS, KatoIII, NUGC3, GCIY, H-111-TC, SH-10-TC, KKLS, GCR1 and the like. Can be done. Gastric cancer cell lines are available from the Human Science Promotion Foundation Research Resource Bank (Osaka, Japan) and the like.

Gastric cancer cell lines MKN45, NUGC4, GCIY, GCR1, MKN7 and OCUM-1 were cultured for 72 hours with 10-200 ng / mL HGF (hepatocyte proliferation factor) and found in the diffuse gastric cancer cell lines NUGC4 and GCIY at baseline. It has been reported that cell proliferation was confirmed to be more than twice the proliferation, but cell proliferation was not confirmed in the non-diffuse gastric cancer cell lines MKN45, GCR1, MKN7 and OCUM-1 (Non-Patent Document 1). When the diffuse gastric cancer cell line NUGC4 was cultured with 100 ng / mL amphiregulin for 72 hours, cell proliferation was confirmed to be about twice that from baseline proliferation, and it was cultured with 1 ng / mL HB-EGF for 72 hours. As a result, it has also been reported that a slight induction of cell proliferation was confirmed (Non-Patent Document 2). In addition, baseline growth refers to the growth of cancer cell lines cultured in the absence of cytokines, growth factors, and the like.

In addition, the NUGC4 cell line is rapidly disseminated intraperitoneally in nude mice by intraperitoneal injection to form bloody ascites (Nakashio T et al., Int J Cancer 1997; 70: 612-8).

HGF has been reported to be overexpressed in many gastric cancers and an increase in serum HGF is associated with poor prognosis (Amemiya et al., Oncology 2002; 63: 286-96, Park et. al., APMIS 2000; 108: 195-200). It has been clarified that HGF is highly produced from cancer stromal fibroblasts that are specifically abundant in diffuse gastric cancer (Non-Patent Document 1). These cancer stroma-derived HGFs cause a strong cell proliferation effect in addition to the hypermotility of diffuse gastric cancer cells via MET, which is the only receptor (Non-Patent Document 1). Furthermore, these paraclin-induced HGFs induce the production of ampphiregulin (Non-Patent Document 2), which is one of the EGFR ligands having remarkable cell proliferation activity like HGF, in a larger amount than diffuse gastric cancer cells (Non-patent). Document 1). HGF-induced and autocrine amphiregulin proliferates diffuse gastric cancer cells by an autocrine mechanism.

On the other hand, in non-diffuse gastric cancer, the induction of HGF production from cancer stromal fibroblasts, which is seen in the cancer stromal of diffuse gastric cancer, is not observed. In addition, non-diffuse gastric cancer cells do not have cell proliferation-inducing effects stimulated by HGF and ampphiregulin, which is a major difference from diffuse gastric cancer cells (Non-Patent Document 1). In diffuse gastric cancer, activation of cancer interstitial-induced paraclinic HGF via the MET receptor (HGF-MET axis pathway) is considered to play an important role (Non-Patent Document 1), and MET. It has been reported that gastric cancer cells with gene amplification are also associated with the development of scirrhous gastric cancer and diffuse gastric cancer (Nakajima M et al., Cancer 1999; 85: 1894-1902).

The interaction of tumor cells with stromal cells plays an important role in tumor development and progression, and stromal fibroblasts are often known to be associated with cancer progression (Kalluri R et al. , Nat Rev Cancer 2006; 6: 392-401). Scirrhous gastric cancer is characterized by rapid diffuse infiltration and proliferation of cancer cells, as well as a high degree of interstitial fibrosis.

(Therapeutic agent 2)
The present invention provides a therapeutic agent for diffuse gastric cancer containing a VEGF receptor 2 inhibitor for use in combination therapy with a MET inhibitor. Furthermore, the present invention provides a therapeutic agent for diffuse gastric cancer containing a MET inhibitor for use in combination therapy with a VEGF receptor 2 inhibitor.

Examples of the MET inhibitor include sabolitinib (AZD6094, HMPL-504), crizotinib (PF-02341066), foretinib, PHA-665752, SU11274, SGX-523, BMS-777607, JNJ-38877605, tivantinib (ARQ197). 04217903, capmatinib (INCB28060), BMS-754807, AMG-208, MK-2461, AMG-458, NVP-BVU972, norcantharidin, AMG337, S49076, NPS-1034 and Merestinib (LY2801653). Inhibitors may include pharmaceutically acceptable salts of these listed. In one embodiment, the MET inhibitor is sabolitinib (AZD6094, HMPL-504).

Savorytinib (AZD6094, HMPL-504) is 1H-1,2,3-triazolo [4,5-b] pyrazine, 1-[(1S) -1-imidazole [1,2-a] pyridine-6-ylethyl ] -6- (1-Methyl-1H-pyrazole-4-yl)-(CAS Registry Number: 1313725-88-0). Crizotinib (PF-02341066) is 3-((R) -1- (2,6-dichloro-3-fluorophenyl) ethoxy) -5- (1- (piperidine-4-yl) -1H-pyrazole-4). -Il) Pyridine-2-amine (CAS Registry Number: 877399-52-5). Foretinib is N- (3-fluoro-4-((6-methoxy-7- (3-morpholinopropoxy) quinoline-4-yl) -oxy) phenyl) -N- (4-fluorophenyl) cyclopropane-1. , 1-Dicarboxamide (CAS Registry Number: 849217-64-7). PHA-665752 is (R, Z) -5- (2,6-dichlorobenzylsulfonyl) -3-((3,5-dimethyl-4- (2- (pyrrolidin-1-ylmethyl) pyrrolidine-1-carbonyl). ) -1H-Pyrrole-2-yl) Methylene) Indoline-2-one (CAS Registry Number: 477575-56-7). SU11274 is (Z) -N- (3-chlorophenyl) -3-((3,5-dimethyl-4- (1-methylpiperazin-4-carbonyl) -1H-pyrrole-2-yl) methylene) -N. -Methyl-2-oxoindoline-5-sulfonamide (CAS Registry Number: 658084-23-2). SGX-523 is a 6- (6- (1-methyl-1H-pyrazol-4-yl)-[1,2,4] triazolo [4,3-b] pyridazine-3-ylthio) quinoline (CAS Registry Number). : 1022150-57-7).

BMS-777607 is N- (4- (2-amino-3-chloropyridin-4-yloxy) -3-fluorophenyl) -4-ethoxy-1- (4-fluorophenyl) -2-oxo-1, 2-Dihydropyridine-3-carboxamide (CAS Registry Number: 1025720-94-8). JNJ-38877605 is a 6- (difluoro (6- (1-methyl-1H-pyrazol-4-yl)-[1,2,4] triazolo [4,3-b] pyridazine-3-yl) methyl) quinoline. (CAS registry number: 943540-75-8). Tivantinib (ARQ197) is (3R, 4R) -3- (2,3-dihydro-1H-pyrrolidine [3,2,1-ij] quinoline-6-yl) -4- (1H-indole-3-yl). ) Pyrrolidine-2,5-dione (CAS Registry Number: 905854-02-6). PF-04217903 is 2- (4-(3- (quinoline-6-ylmethyl) -3H- [1,2,3] triazolo [4,5-b] pyrazine-5-yl) -1H-pyrazole-1. -Il) Ethanol (CAS Registry Number: 956905-27-4). Capmatinib (INCB28060) is 2-fluoro-N-methyl-4- (7- (quinoline-6-ylmethyl) imidazole [1,2-b] [1,2,4] triazine-2-yl) benzamide (CAS). Registration number: 1029712-80-8). BMS-754807 is a combination of (S) -1- (4- (5-cyclopropyl-1H-pyrazole-3-ylamino) pyrrolo [1,2-f] [1,2,4] triazine-2-yl)-. N- (6-fluoropyridin-3-yl) -2-methylpyrrolidine-2-carboxamide (CAS Registry Number: 1001350-96-4).

AMG-208 is a 7-methoxy-4-((6-phenyl- [1,2,4] triazolo [4,3-b] pyridazine-3-yl) methoxy) quinoline (CAS Registry Number: 1002304-34-). 8). MK-2461 is N-((2R) -1,4-dioxane-2-ylmethyl) -N-methyl-N'-[3-(1-methyl-1H-pyrazole-4-yl) -5-oxo. -5H-benzo [4,5] cycloheptane [1,2-b] pyridin-7-yl] sulfamide (CAS Registry Number: 917879-39-1). AMG-458 is 1- (2-hydroxy-2-methylpropyl) -N- (5- (7-methoxyquinoline-4-yloxy) pyridin-2-yl) -5-methyl-3-oxo-2- Phenyl-2,3-dihydro-1H-pyrazole-4-carboxamide (CAS Registry Number: 913376-83-7). NVP-BVU972 is a 6-((6- (1-methyl-1H-pyrazol-4-yl) imidazole [1,2-b] pyridazine-3-yl) methyl) quinoline (CAS Registry Number: 1185763-69-). 2). Norcantharidin is exo-3,6-epoxyhexahydrophthalic anhydride (CAS Registry Number: 29745-04-8).

AMG337 is 1,6-naphthylidine-5 (6H) -on, 6-[(1R) -1- [8-fluoro-6- (1-methyl-1H-pyrazole-4-yl) -1,2, 4-Triazolo [4,3-a] Pyridine-3-yl] Ethyl] -3- (2-Methoxyethoxy)-(CAS Registry Number: 1173699-31-4). S49076 contains 2,4-thiazolidinedione, 3-[[2,3-dihydro-3-[[4- (4-morpholinylmethyl) -1H-pyrrole-2-yl] methylene] -2-oxo- 1H-Indole-5-yl] Methyl]-(CAS Registry Number: 1265965-22-7). NPS-1034 is N- (3-fluoro-4- (3-phenyl-1H-pyrolo [2,3-b] pyridin-4-yloxy) phenyl) -2- (4-fluorophenyl) -1,5. -Dimethyl-3-oxo-2,3-dihydro-1H-pyrazole-4-carboxamide (CAS Registry Number: 1221713-92-3). Mrestinib (LY2801653) is 3-pyridinecarboxamide, N- [3-fluoro-4-[[1-methyl-6- (1H-pyrazole-4-yl) -1H-indazole-5-yl] oxy] phenyl]. It is -1- (4-fluorophenyl) -1,2-dihydro-6-methyl-2-oxo- (CAS Registry Number: 1206799-15-6).

The VEGF receptor 2 inhibitor may be an anti-VEGF receptor 2 antibody, and any antibody that binds to VEGF receptor 2 can be used. In one embodiment, the anti-VEGF receptor 2 antibody is ramucirumab. Ramucirumab is marketed as Cyramza® and is also known as IMC-1121B or CAS Registry Number 947687-13-0. Ramucirumab inhibits the binding of VEGF to the vascular endothelial growth factor receptor (VEGFR2) and inhibits angiogenesis in tumor tissue. The anti-VEGF receptor 2 antibody DC101 is a rat monoclonal antibody against mouse VEGF receptor 2 that can be used experimentally as a substitute for anti-VEGF receptor 2 antibody, preferably ramucirumab, in mice (Witte L., et al Cancer Metastasis). Rev., 17, 155-161, 1998).

In combination therapy, two or more agents (specifically, the MET inhibitor and the VEGF receptor 2 inhibitor) can be administered simultaneously or sequentially. When two or more drugs are administered simultaneously or sequentially, all the drugs may be administered by the same route or by different routes.

As used herein, co-administration means that two or more agents are administered at intervals of minutes to seconds or less. For example, the two drugs are administered at intervals of about 15 minutes to 1 minute or less. As used herein, sequential administration means administration at intervals of minutes, hours, days, weeks. For example, two drugs are available for 15 minutes or more, 30 minutes or more, 60 minutes or more, or 1 day, 2 days, 3 days, 4 days, 5 days, 6 days or 7 days, or 2 weeks, 3 weeks or 4 Administered over a week.

In combination therapy, the MET inhibitor can be co-administered or sequentially administered with the VEGF receptor 2 inhibitor. In combination therapy, the VEGF receptor 2 inhibitor can be co-administered or sequentially administered with the MET inhibitor.

In one embodiment, the MET inhibitor contained in the therapeutic agent of the present invention is saboritinib, which can be used in combination therapy with a VEGF receptor 2 inhibitor. In another embodiment, the VEGF receptor 2 inhibitor contained in the therapeutic agent of the present invention is ramucirumab, which can be used in combination therapy with a MET inhibitor.

Saboritinib can be orally administered to adults at 100 mg to 1200 mg, preferably 300 mg to 1200 mg, and more preferably 300 mg to 600 mg per day in combination with the anti-VEGF receptor 2 antibody, 1 to 3 times per day. Can be divided into two doses. In combination with a MET inhibitor, ramucirumab can be infused once every two weeks for adults, and 8 mg / kg (body weight) of ramucirumab once for about 60 minutes.

The pharmaceutically acceptable salt of the compound contained in the therapeutic agent of the present invention includes an acid addition salt and a base addition salt (including a disalt) thereof. Suitable acid addition salts are formed from acids that form non-toxic salts. For example, acetate, asparagate, benzoate, besilate, bicarbonate / carbonate, bicarbonate, camsilate, citrate, edicylate, esylate, fumarate, gluceptate. , Gluconate, glucuronic acid, hibenate, hydrochloride / chloride, hydrobromide / bromide, hydroiodide / iodide, hydrogen phosphate, D and L-lactate, malate , Maleate, malonate, mesylate, methylsulfate, 2-naptylate, nicotinate, nitrate, orotate, palmitate, phosphate, saccharate, stearate, succinate Acids, D- and L-tartrates, and tosylate salts. Suitable base salts are formed from bases that form non-toxic salts. Examples include: aluminum, ammonium, arginine, benzathine, calcium, choline, diethylamine, diolamine, glycine, lysine, magnesium, meglumine, olamine, potassium, sodium, tromethamine and zinc salts.

The therapeutic agent provided by the present invention is prescribed in an optimal dosage form according to the route of administration. The therapeutic agent may be a solid preparation such as a tablet, a granule, a fine granule, a powder or a capsule, or an oral preparation such as a jelly or syrup, and an injection, a suppository, an ointment, etc. It may be a preparation for parenteral administration such as a suppository.

The therapeutic agents provided by the present invention may contain pharmaceutically acceptable additives. Examples of the pharmaceutically acceptable additive include tonicity agents, pH adjusters, buffers, stabilizers, freeze-protectors, antibiotics and the like. Specific examples thereof include water, ethanol, sodium chloride, glucose, albumin and the like. Further, as another example of a pharmaceutically acceptable additive, an excipient, a binder, a lubricant and the like can be exemplified. Examples of the excipient include lactose, cornstarch, sucrose, glucose, sorbitol, crystalline cellulose, silicon dioxide and the like. Examples of the binder include polyvinyl alcohol, ethyl cellulose, methyl cellulose, gum arabic, hydroxypropyl cellulose, hydroxypropyl methyl cellulose and the like. Examples of the lubricant include magnesium stearate, talc, silica and the like.

The production of the therapeutic agent of the present invention is preferably carried out under conditions conforming to the manufacturing control and quality control rules for pharmaceutical products and quasi-drugs (good manufacturing practice, GMP).

The therapeutic agent of the present invention can be used to treat diffuse gastric cancer in a patient. In another aspect, the therapeutic agent of the present invention can be used to treat scirrhous gastric cancer in a patient. If the patient with diffuse gastric cancer has developed cancerous peritonitis, it can be used to suppress the exacerbation of the symptoms of cancerous peritonitis, or to relieve or cure the symptoms. In addition, the therapeutic agent of the present invention can be used to prevent the onset of cancerous peritonitis when the patient with the above-mentioned diffuse gastric cancer does not develop cancerous peritonitis. As used herein, cancerous peritonitis refers to a condition in which cancer has spread to the peritoneum, and cancer cells are disseminated into the abdominal cavity to form a large number of nodules of cancer cells on the peritoneum (peritoneal dissemination). ). Cancerous peritonitis is often accompanied by malignant ascites with increased peritoneal dissemination. Cancerous peritonitis is also called peritoneal peritonitis. The therapeutic agent of the present invention can be used for treating diffuse gastric cancer having cancerous peritonitis. The therapeutic agents of the present invention can be used to treat diffuse gastric cancer with unresectable or metastatic disease. The therapeutic agent of the present invention can be used to suppress ascites production in patients with diffuse gastric cancer. The therapeutic agent of the present invention can be used to prevent ascites production when a patient with diffuse gastric cancer is not in a state of ascites retention.

(Method of treatment)
The present invention is a method of treating diffuse gastric cancer in a patient by suppressing the activity of MET and VEGF receptor signaling pathways.
Provided are methods comprising administering to the patient one or more agents for suppressing the activity of the MET and VEGF receptor signaling pathways.

In one embodiment, the method of treating diffuse gastric cancer in a patient by inhibiting the activity of the MET and VEGF receptor signaling pathways of the invention comprises applying a double inhibitor of MET and VEGF receptor 2 to the patient. It can be administered or by administering to the above-mentioned patients a combination therapy of a MET inhibitor and a VEGF receptor 2 inhibitor.

According to the treatment method of the present invention, the survival time is statistically significantly extended as compared with the control group (vehicle control). By the therapeutic method of the present invention, EGF is used with an EGF receptor inhibitor (HER1 / 4-TKI) and an anti-VEGF receptor 2 antibody by doubly inhibiting the signal transduction pathway of MET and VEGF receptor 2. Survival is prolonged rather than blocking the signaling pathways of the receptor and VEGF receptor 2.

In the examples described below, the treatment method of the present invention has a remarkable survival at least during the treatment period (drug administration period) and the first half of the drug holiday as compared with the monotherapy of each of the MET inhibitor and the anti-VEGFR2 antibody. It showed an improvement in the rate. When the data shown in FIGS. 1 and 2 are collectively analyzed, the survival rate of the sabolitinib group (MET inhibitor) was 21% (3 out of 14 animals survived) at the end of the treatment period (7 weeks after gastric cancer cell transplantation). ), And the survival rate of the anti-VEGFR2 antibody group is 42% (6 out of 14 animals survive), but the survival rate of the combination therapy of the MET inhibitor and the anti-VEGFR2 antibody is 100% (12 out of 12 animals). Survival), and the combination therapy of MET inhibitor and anti-VEGFR2 antibody showed higher effect than the addition of each monotherapy. On the other hand, in the combination therapy of HER1 / 4-TKI (AZ5104 or osimertinib) and anti-VEGFR2 antibody, the survival rates are 67% and 50% (AZ5104 or osimertinib) at the end of the treatment period (7 weeks after gastric cancer cell transplantation). It is a combination therapy with anti-VEGFR2 antibody), and no difference from anti-VEGFR2 antibody monotherapy (50%) was observed (Fig. 1).

In the examples described below, mice transplanted with diffuse gastric cancer cells were used as a control group in a treatment group that inhibited MET and VEGF receptor 2 (golvatinib high-dose group, cabozantinib group, savorytinib and anti-VEGFR2 antibody combination group). On the other hand, a good life-prolonging effect was observed (Fig. 2). It was suggested that suppression of the activity of MET and VEGF receptor signaling pathways would result in a therapeutic effect on diffuse gastric cancer in patients. Even with a dual inhibitor of MET and VEGF receptor 2 (eg, gorbatinib, cabozantinib), a combination therapy of a MET inhibitor (eg, sabolitinib) and a VEGF receptor 2 inhibitor (eg, anti-VEGFR2 antibody) is used. However, a good life-prolonging effect was observed.

In the examples described below, in mice in which diffuse gastric cancer cells are transplanted intraperitoneally and exhibit ascites retention, by doubly inhibiting the signaling pathway of MET and VEGF receptor 2, 3 to 10 days after the start of dosing. It became clear that the ascites disappeared. Ascites disappearance was most prominent in the group treated with cabozantinib and disappeared 3-4 days after drug administration. No effect of suppressing ascites production was observed with the MET inhibitor alone, and even if the dose was increased, the rate of ascites increase tended to be slower than that of the control group. Mice transplanted with diffuse gastric cancer cells into the abdominal cavity have increased ascites retention and die within 1 to 2 weeks from a tense state if not treated, so suppression of ascites production and disappearance of ascites prolong the survival period. It is suggested that he is deeply involved in. With the combination therapy of MET and VEGF receptor 2 double inhibitor and VEGF receptor 2 inhibitor (eg, anti-VEGFR2 antibody), ascites that had disappeared during drug administration maintained its state without re-reservation. ..

In the therapeutic method of the present invention using a double inhibitor of MET and VEGF receptor 2 or a combination therapy of MET inhibitor and VEGF receptor 2 inhibitor, the therapeutic agent is orally administered, gastrically, by any suitable means. It can be administered intranasally, intranasally, intralesional, or topically. The therapeutic agent of the present invention may be administered by parenteral injection, which includes intramuscular, intravenous, arterial, intraperitoneal, or subcutaneous injection. The dosing resume is appropriately planned depending on whether the dosing is short-term or long-term. Based on the knowledge of the healthcare professional, the optimal and clinically appropriate route of administration, administration method, dose and administration schedule for treatment are determined.

The therapeutic method of the present invention is used in patients in which the expression level of MET and / or VEGF receptor 2 in a patient-derived gastric cancer cell sample is statistically similar to or higher than a predetermined control level. It can be carried out against. The control level may be the average value of MET and / or VEGF receptor 2 expression levels in multiple specimens with diffuse gastric cancer that is sensitive to MET and / or VEGF receptor 2 inhibitors.

The therapeutic method of the present invention can be performed on a patient in which MET and / or VEGF receptor 2 is phosphorylated or not phosphorylated in a patient-derived gastric cancer cell sample. In one embodiment, the therapeutic method of the invention can be selectively performed on a patient to whom MET and / or VEGF receptor 2 is phosphorylated in a patient-derived gastric cancer cell sample. The above-mentioned double inhibitor of MET and VEGF receptor 2, or combination therapy of MET inhibitor and VEGF receptor 2 inhibitor has inhibitory activity on phosphorylated MET and / or VEGF receptor 2. May be.

The therapeutic method of the present invention may be applied to a patient as a single therapy with a double inhibitor of MET and VEGF receptor 2, or in addition to the double inhibitor of MET and VEGF receptor 2, conventional surgery or It may be accompanied by radiation therapy, chemotherapy, antibody drugs, immunotherapy. Such chemotherapeutic or antibody agents can be administered simultaneously, sequentially or separately, with treatment with a double inhibitor of MET and VEGF receptor 2. The same applies to the combination therapy of the MET inhibitor and the VEGF receptor 2 inhibitor.

In the therapeutic method of the present invention, the chemotherapy used in combination with the double inhibitor of MET and VEGF receptor 2 includes oxaliplatin, capecitabine, fluorouracil, irinotecan, cisplatin, docetaxel, paclitaxel, tegafur, doxiflulysin, and tegafur / uracil. It can be selected from anticancer agents such as agents and anticancer agents containing tegafur, gimeracil, and oteracil potassium. The same applies to the combination therapy of the MET inhibitor and the VEGF receptor 2 inhibitor.

In the therapeutic method of the present invention, the antibody drugs used in combination with the double inhibitor of MET and VEGF receptor 2 include anti-PD-1 antibody nivolumab, anti-PD-1 antibody pembrolizumab, anti-PD-L1 antibody avelumab, and anti-HER2. It can be selected from, but is not limited to, anti-cancer agents such as the antibody pembrolizumab. The same applies to the combination therapy of the MET inhibitor and the VEGF receptor 2 inhibitor.

The embodiments of the present invention described below are for illustration purposes only and do not limit the technical scope of the present invention. The technical scope of the invention is limited only by the description of the claims. Modifications of the present invention, for example, addition, deletion and replacement of the constituent elements of the present invention may be made on condition that the gist of the present invention is not deviated.

In vivo test AZ5104, osimertinib (AZD9291), sabolitinib (AZD6094), METTKI (MET tyrosine kinase inhibitor, PF-04217903), cabozantinib (BMS-907351) and gorbatinib (E7050) were obtained from Selleck Chemicals. The anti-VEGFR2 antibody is an anti-mouse VEGF receptor 2 antibody DC101 (GeneTex).

Diffuse gastric cancer cells ( ^NUGC4 cells) were adjusted to 200 μL of phosphate buffered saline (PBS) per animal in Balb / c nu / nu mice (6 weeks old) and transplanted by intraperitoneal administration. On the 21st day after transplantation, ascites retention was confirmed and administration of the drug was started.

A. Comparison of combination therapy "HER1 / 4-TKI + anti-VEGFR2 antibody" and "MET-TKI + anti-VEGFR2 antibody" The treatment was performed in the following groups, and 6 mice were used in each treatment group.
(1) AZ5104 group single dose 10 mg / Kg (body weight) once daily administration (2) ossimertinib group single dose 10 mg / Kg (body weight) once daily administration (3) sabolitinib group single dose 2. 5 mg / Kg (body weight) once daily administration (4) Anti-VEGFR2 antibody group Single dose 40 mg / Kg (body weight) twice weekly (5) AZ5104 and anti-VEGFR2 antibody combination group AZ5104 single dose 10 mg / Combination of Kg (body weight) once daily and anti-VEGFR2 antibody single dose 40 mg / Kg (body weight) twice weekly (6) Combination group of osimertinib and anti-VEGFR2 antibody Oshimeltinib single dose 10 mg / Kg ( Combination of once-daily administration of anti-VEGFR2 antibody 40 mg / Kg (body weight) twice weekly (7) Combination group of saboritinib and anti-VEGFR2 antibody Savorytinib single dose 2.5 mg / Kg (body weight) Body weight) and anti-VEGFR2 antibody single dose 40 mg / Kg (body weight) twice weekly combination AZ5104, osimertinib, savorytinib were administered by the oral route, and anti-VEGFR2 antibody was administered intraperitoneally.
Only the solution without drug dissolution was administered once a week to the control group (vehicle control). Administration was continued for 4 weeks (treatment period), followed by a 4-week drug holiday observation period.

Results: Ascites of mice 21 days after gastric cancer cell transplantation was in a mild to moderate retention state. In the control group, all mice died within 1 to 2 weeks (within 5 weeks after gastric cancer cell transplantation) after the ascites increased and became tense.

Seven weeks after gastric cancer cell transplantation, 2 animals (33%) in the AZ5104 group, 0 animals (0%) in the osimertinib group, 1 animal (17%) in the savorytinib group, and 3 animals (50%) in the anti-VEGFR2 antibody group. 4 (67%) survived in the AZ5104 plus anti-VEGFR2 antibody group, 3 (50%) in the ossimertinib plus anti-VEGFR2 antibody combination group, and 6 (100%) survived in the savorytinib plus anti-VEGFR2 antibody combination group. Was there. Furthermore, in the combination group of sabolitinib and anti-VEGFR2 antibody, all mice (100%) survived until 8 weeks after gastric cancer cell transplantation.

FIG. 1 shows a survival curve. The survival curve was created by the Kaplan-Meier method, and the difference between the two groups of survival curves was tested by the log rank test. As a result of performing a log rank test against the control group for survival rate, all the combination groups tested (that is, the combination group of AZ5104 and anti-VEGFR2 antibody, the combination group of osimertinib and anti-VEGFR2 antibody, and the combination group of sabolitinib and anti-VEGFR2 antibody). There was a significant difference in the group) (P <0.05).

Combination therapy with the MET inhibitor and anti-VEGFR2 antibody was observed to significantly improve survival, at least during the treatment period and the first half of the drug holiday, compared to the respective monotherapy. At the end of the treatment period (7 weeks after gastric cancer cell transplantation), the survival rate of the savorytinib group (MET inhibitor) was 17% and that of the anti-VEGFR2 antibody group was 50%, but that of the MET inhibitor and anti-VEGFR2. The survival rate for combination therapy with antibodies is 100%. On the other hand, in the combination therapy of HER1 / 4-TKI (AZ5104 or osimertinib) and anti-VEGFR2 antibody, the survival rates are 67% and 50% (AZ5104 or osimertinib) at the end of the treatment period (7 weeks after gastric cancer cell transplantation). It is a combination therapy with anti-VEGFR2 antibody), and no difference from anti-VEGFR2 antibody monotherapy (50%) was observed.

B. Comparison of multiple inhibitors (cabozantinib, gorbatinib) and combination therapy "MET inhibitor + VEGFR inhibitor" The treatment was performed in the following groups, and 6 mice were used in each treatment group.
(1) Sabolitinib group A single dose of 2.5 mg / Kg (body weight) was administered once a day.
(2) PF-04217903 group A single dose of 24 mg / Kg (body weight) was administered once a day.
(3) Anti-VEGFR2 antibody group A single dose of 40 mg / Kg (body weight) was administered twice a week.
(4) Golbatinib low-dose group A single dose of 25 mg / Kg (body weight) was administered once a day.
(5) Golbatinib high-dose group A single dose of 50 mg / Kg (body weight) was administered once a day.
(6) Cabozantinib group A single dose of 30 mg / Kg (body weight) was administered once a day.
(7) Combination group of sabolitinib and anti-VEGFR2 antibody: A combination of a single dose of sabolitinib 2.5 mg / Kg (body weight) once daily and an anti-VEGFR2 antibody single dose 40 mg / Kg (body weight) twice a week. ,
Savorytinib, PF-04217903, gorbatinib and cabozantinib were administered by the oral route, and anti-VEGFR2 antibody was administered intraperitoneally.
Only the solution without drug dissolution was administered once a week to the control group (vehicle control). Administration was continued for 4 weeks, followed by a 4-week washout observation period.
Ascites retention was performed by visual observation and palpation of the abdomen. The accumulation of ascites was confirmed by the swelling of the abdomen and the vibration during palpation. In addition, it was judged that ascites had disappeared because the abdomen was flattened and there was no vibration during palpation.

Results: Ascites of mice 21 days after gastric cancer cell transplantation was in a mild to moderate retention state.
Control group: All mice died within 1 to 2 weeks (within 5 weeks after gastric cancer cell transplantation) after the ascites increased and became tense.
Sabolitinib group: No ascites-suppressing effect was observed by drug administration, and the patient died within 1 to 2 weeks after becoming tense after increasing the dose.
Anti-VEGFR2 antibody group: Ascites gradually decreased from 7 to 10 days after the start of drug administration (treatment), and then remained in a reduced state for about 1 week. did. After that, he became tense over a few weeks and died.
Combination group of sabolitinib and anti-VEGFR2 antibody: Ascites disappeared on the 10th day from the start of drug administration (treatment), and the disappearance state was maintained for 2 weeks thereafter. Ascites re-accumulation was confirmed 7 to 3 days before the end of the treatment period, and then slowly became tense and died over 3 to 4 weeks.
Cabozantinib group: Ascites disappeared 3 to 4 days after the start of drug administration (treatment), and ascites retention appeared from 3 to 4 days after the end of drug administration (after the end of the treatment period). After that, within 1 to 2 weeks, the ascites increased and he died in a tense state.
High-dose group of gorbatinib: Ascites disappeared on the 7th day after the start of drug administration (treatment), and ascites re-retention appeared on the 10th day after the end of the drug administration (after the end of the treatment period). Died from.

Four (67%) mice in the gorbatinib high-dose group, two (34%) in the cabozantinib group, and one (17%) in the savorytinib plus anti-VEGFR2 antibody group survived 11 weeks after gastric cancer cell transplantation. )Met. Seven weeks after gastric cancer cell transplantation, these groups consisted of 6 animals (100%) in the gorbatinib high-dose group, 5 animals (83%) in the cabozantinib group, and 6 animals (100%) in the combination group of sabolitinib and anti-VEGFR2 antibody. Was alive.
Seven weeks after gastric cancer cell transplantation, 4 (67%) in the gorbatinib low dose group, 4 (50%) in the PF-0427903 group, 3 (38%) in the anti-VEGFR2 antibody group, and 2 in the sabolitinib group ( 25%) were alive, but all mice in these groups died 11 weeks before gastric cancer cell transplantation.

FIG. 2 shows the survival curve. The survival curve was created by the Kaplan-Meier method, and the difference between the two groups of survival curves was tested by the log rank test. As a result of log rank test for the control group, there was a significant difference in survival rate between the high dose group of gorbatinib, the low dose group of gorbatinib, the cabozantinib group, the combination group of sabolitinib and anti-VEGFR2 antibody, and the anti-VEGFR2 antibody group ( P <0.05).

Mice transplanted with diffuse gastric cancer cells had a good life extension compared to the control group in the treatment group that inhibited MET and VEGF receptor 2 (golvatinib high dose group, cabozantinib group, savorytinib and anti-VEGFR2 antibody combination group). The effect was seen. It is suggested that suppression of the activity of MET and VEGF receptor signaling pathways results in a therapeutic effect on diffuse gastric cancer in patients.

Claims (12)

A therapeutic agent for diffuse gastric cancer, comprising a double inhibitor of MET and VEGF receptor 2. Double inhibitors of MET and VEGF receptor 2 are cabozantinib (BMS-907351), cabozantinib malate (XL184), gorbatinib (E7050), MGCD-265, BMS-794833 and Nintinib (CT-053, DE-). The therapeutic agent according to claim 1, which is selected from the group consisting of 120, CT053PTSA). A therapeutic agent for diffuse gastric cancer containing a VEGF receptor 2 inhibitor for use in combination therapy with a MET inhibitor. A therapeutic agent for diffuse gastric cancer containing a MET inhibitor for use in combination therapy with a VEGF receptor 2 inhibitor. The therapeutic agent according to claim 3 or 4, wherein in the combination therapy, the MET inhibitor and the VEGF receptor 2 inhibitor are co-administered or sequentially administered. The MET inhibitors include sabolitinib (AZD6094, HMPL-504), crizotinib (PF-02341066), foretinib, PHA-665752, SU11274, SGX-523, BMS-777607, JNJ-388777605, tivantinib (ARQ197), F. , Capmatinib (INCB28060), BMS-754807, AMG-208, MK-2461, AMG-458, NVP-BVU972, Norcantharidin, AMG337, S49076, NPS-1034 and Merestinib (LY2801653). The therapeutic agent according to any one of claims 3 to 5, which is sabolitinib (AZD6094, HMPL-504). The therapeutic agent according to any one of claims 3 to 6, wherein the VEGF receptor 2 inhibitor is an anti-VEGFR2 antibody, preferably ramucirumab. The therapeutic agent according to any one of claims 1 to 7, wherein the diffuse gastric cancer is scirrhous gastric cancer. The therapeutic agent according to any one of claims 1 to 8, wherein the diffuse gastric cancer has cancerous peritonitis. The therapeutic agent according to any one of claims 1 to 9, wherein the diffuse gastric cancer has unresectable or metastatic disease. A method of treating diffuse gastric cancer in patients by suppressing the activity of MET and VEGF receptor signaling pathways.
A method comprising administering to said patient one or more agents for suppressing the activity of the MET and VEGF receptor signaling pathways. 11. The method of claim 11, comprising administering to the patient a dual inhibitor of MET and VEGF receptor 2 or a combination therapy of a MET inhibitor and a VEGF receptor 2 inhibitor to the patient. ..

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