CN109553581B

CN109553581B - Substituted urea compounds, pharmaceutically acceptable salts or solvates thereof, uses thereof, medicaments and pharmaceutical compositions

Info

Publication number: CN109553581B
Application number: CN201910114715.2A
Authority: CN
Inventors: 陆瑞燕
Original assignee: Guangzhou Liushun Biotechnology Co ltd
Current assignee: Guangzhou six Shun biological Polytron Technologies Inc
Priority date: 2018-09-25
Filing date: 2019-02-14
Publication date: 2022-01-25
Anticipated expiration: 2039-02-14
Also published as: CN109553581A

Abstract

The present invention provides a substituted urea compound, a pharmaceutically acceptable salt thereof, or a solvate thereof. The invention also provides application of the substituted urea compound, the pharmaceutically acceptable salt or the solvate thereof as an inhibitor of one of MET, VEGFR, PDGFR and RET or application of the substituted urea compound, the pharmaceutically acceptable salt or the solvate thereof in preparation of a multi-target inhibitor of two or more of MET, VEGFR, PDGFR and RET, and application of the substituted urea compound, the pharmaceutically acceptable salt or the solvate thereof in preparation of a drug for regulating kinase activity or treatingUse in a medicament or pharmaceutical composition for the treatment of a kinase-associated disease. The invention also provides a medicament or a pharmaceutical composition for treating the angiogenesis abnormal diseases. The structure of the substituted urea compound is shown as a formula (I), R₁Selected from methoxy or deuterated methoxy; r₂Selected from methyl, deuterated methyl,

Any one of the structures of (1); r₃Is selected from any one of F, Cl, Br and I.

Description

Substituted urea compounds, pharmaceutically acceptable salts or solvates thereof, uses thereof, medicaments and pharmaceutical compositions

RELATED APPLICATIONS

The priority of the chinese patent application entitled "substituted urea compounds, pharmaceutically acceptable salts thereof, or solvates thereof, pharmaceutical compositions, methods of preparation, and uses thereof," filed on 25/09/2018, having application number 2018111165044, is hereby incorporated by reference in its entirety.

Technical Field

The invention belongs to the technical field of biological medicines, and relates to a substituted urea compound, pharmaceutically acceptable salt or solvate thereof, application thereof, a medicine and a pharmaceutical composition.

Background

Cancer is a series of diseases characterized by uncontrolled proliferation and spread of abnormal cells, and is one of the major diseases seriously harming human health, and according to the report of the world health organization, the number of cancer patients increases by 1000 million every year, about 700 million people die, and by 2020, 2000 million people will be newly added to cancer patients every year. Currently, 160 million people suffer from cancer every year in our country, and at the same time, about 130 million people die from cancer, which is becoming the second killer of people who are second only to cardiovascular diseases.

With the development of molecular biology, the molecular mechanism of action for the development of tumorigenesis has made a great progress, and it has been now demonstrated that various receptor tyrosine kinase families and their ligands play key regulatory roles in tumor angiogenesis, such as MET, VEGFR, BRAF, PDGFR and RET.

RET is a proto-oncogene, full name: RET proto-oncogene, located in the long arm of autosomal 10 (10q11.2), 60kb in length, comprising 21 exons, encodes an 1100 amino acid tyrosine kinase receptor superfamily RET protein, which is expressed in normal neurons, sympathetic and parasympathetic ganglia, thyroid C cells, adrenal medulla cells, urogenital tract cells, testicular germ cells. Activation of RET proteins activates downstream signaling pathways (including RAS, MAPK, ERK, PI3K, AKT, etc.) leading to cell proliferation, migration, and differentiation. The RET gene activating mutation is related to human malignant tumors including multiple endocrine adenomatosis type 2 (MEN 2), Papillary Thyroid Carcinoma (PTC), congenital megacolon and lung adenocarcinoma, etc., and different mutation types thereof can cause different tumor invasion abilities. Preliminary studies indicate that it mediates a unique signal transduction pathway. The 3 mutant types of RET gene are associated with the development of various cancers in humans: 1. papillary thyroid adenocarcinomas have multiple rearrangements of the RET gene with other genes; 2.7 site point mutations exist in multiple endocrine adenoma 2 types, familial hereditary medullary thyroid carcinoma and the like; 3. lung cancer RET gene fusion. In recent years, studies on the influence of mutations in the RET gene on the function of RET proteins have been focused on the structural functions of RET proteins. With the intensive research on RET gene mutation, RET can become a target for precise treatment of various tumors.

Vascular Endothelial Growth Factor Receptors (VEGFR) are a class of tyrosine kinase transmembrane glycoproteins, which are extracellular domains composed of 7 Ig-like domains; a transmembrane domain and intracytoplasmic tyrosine kinase domain, including 3 receptors for VEGFR-1(Flt-1), VEGFR-2(KDR/Flk-1), VEGFR-3 (Flt-4). The extracellular domain of VEGFR is the region that binds VEGF, which undergoes conformational changes upon binding, leading to receptor dimerization, and autophosphorylation of its intracellular tyrosine site, which activates downstream signaling pathways. VEGFR-1 is mainly distributed in vascular endothelial cells, is also expressed in macrophages, monocytes and hematopoietic stem cells, and can be combined with VEGF-A, VEGF-B and PlGF-1. VEGFR-1 has 10-fold higher affinity for VEGF than VEGFR-2, but does not activate signals for endothelial cell proliferation. VEGFR-1 may act as a reverse regulator of VEGF angiogenesis, and VEGFR-1 may also have biological functions such as mediating monocyte migration, recruitment of endothelial progenitor cells, and promoting survival of hematopoietic stem cells. VEGFR-2 is distributed primarily in vascular endothelial cells and hematopoietic stem cells, but is also expressed in some non-endothelial cell types. VEGFR-2 binds to VEGF-A, VEGF-C, VEGF-D, VEGF-E. The effects of VEGF in stimulating endothelial cell proliferation, increasing vascular permeability and neovascularization are primarily achieved by binding to and activating VEGFR-2. VEGFR-3 is expressed mainly in lymphatic endothelial cells, but also in monocytes, macrophages and the like. VEGFR-3 may bind to VEGF-C and VEGF-D. VEGFR-3 is involved in maintaining the survival of lymphatic endothelial cells and promoting their proliferation and migration, and is involved in tumor cell lymph node metastasis.

Numerous studies have shown that VEGFR is overexpressed in many cancer tissues, including liver, lung, colon, ovary, breast, etc., which play a critical role in tumor growth and metastasis, and therefore can control tumor growth by blocking or interfering with VEGFR signaling pathways. The anti-tumor drug targeting VEGFR has great advantages compared with the traditional tumor treatment drug. Under normal physiological conditions, human angiogenesis only plays a role in physiological activities such as wound healing, menstrual cycle and the like, so that the anti-angiogenesis medicine is used for treating tumors, and has small toxic effect on human bodies; the growth and migration of the tumor depend on the generation of a large number of new blood vessels, and the broad-spectrum anti-tumor effect can be achieved by taking VEGFR as a target spot; unlike tumor cells, vascular endothelial cells have genetic stability and are considered as ideal therapeutic targets, and are not prone to develop resistance to anti-angiogenic drugs; moreover, the direct contact between the endothelial cells and the blood makes the medicine more easily reach the vascular endothelial cells.

In addition, Vascular Endothelial Growth Factor (VEGF) is highly expressed during pathological changes in retinal vascular diseases; clinical trials with anti-VEGF therapy have demonstrated that treatment results in a reduction in choroidal neovascular membranes, reduced fluid leakage, and better efficacy of anti-VEGF therapy in the treatment of neovascular age-related macular degeneration, choroidal neovascular membranes of various etiologies, macular edema due to diabetes and venous obstruction, retinopathy of prematurity, and neovascular glaucoma. Therefore, in addition to tumors, VEGFR inhibitors are also useful in the treatment of related angiogenic disorders such as retinal angiogenesis, neovascular glaucoma, and diabetic retinopathy.

Platelet-derived Growth Factor (PDGF) belongs to the vascular endothelial Growth Factor family, and the activity enhancement of PDGF has important significance in cancer and leukemia, is one of important influencing factors in the initial stage of atherosclerosis, and is the strongest mitogenic Factor of hepatic stellate cells. PDGFR is a tyrosine kinase receptor, has protein tyrosine kinase activity, and after being combined with ligand PDGF, signals are initiated and amplified through dephosphorylation of specific tyrosine residues, actin rearrangement is promoted, and physiological effects such as mitogenesis and chemotaxis are exerted. Overexpression of PDGF and PDGFR families is closely related to a series of diseases such as malignant tumors, atherosclerosis, arterial restenosis, fibrosis and the like, and the PDGF signal transduction is reduced mainly by inhibiting PDGFR and blocking tyrosine kinase phosphorylation and downstream signal transduction of PDGFR. In recent years, research shows that the growth of tumors is angiogenesis-dependent, and tumor cells can produce a plurality of angiogenesis promoting factors and play an important role in the generation, development, invasion and metastasis processes of tumors. PDGF expression is closely related to tumor angiogenesis, tumor cells promote angiogenesis by releasing PDGF, PDGF can also up-regulate the expression of Vascular Endothelial Growth Factor (VEGF), and VEGF is also an important angiogenesis promoting Factor and can indirectly mediate angiogenesis. In vivo and in vitro experimental studies show that the overexpression of PDGF is detected in various tumors such as ovarian cancer, renal cancer, lung cancer, brain tumor, prostatic cancer, breast cancer, colorectal cancer and the like. In addition, activation of PDGF signaling pathway can increase Interstitial Fluid Pressure (IFP) of tumor stroma, and IFP increase is commonly existed in solid tumor tissue, thus seriously preventing the effective transportation of antitumor drug to tumor cells and reducing the drug intake of tumor tissue. At present, PDGFR is mainly used as an anti-tumor target, research and development of PDGFR inhibitors are also the current research focus, and a medicament taking PDGFR as the target has the advantages of high selectivity and small toxic and side effects, but also has the defect that only inhibition is realized and tumor cells are difficult to kill completely.

The size of a protein MET encoded by a mesenchyme to epidermal transformation factor (MET) gene is about 170k Da, and is about 190k Da after glycosylation modification, and finally two polypeptide chains connected by disulfide bonds, namely an alpha chain (50k Da) and a beta chain (140k Da) are formed through shearing action. MET belongs to the RON subfamily and is the only known receptor for scattering factor or Hepatocyte Growth Factor (HGF). The MET protein is a heterodimer consisting of a 50kD subunit only and a 145kD B subunit linked by a disulfide bond, and is divided into an extracellular domain and an intracellular domain. The extracellular domain comprises 3 functionally distinct domains: an N-terminal ligand binding domain (SEMA region) covering the entire ol chain and part of the B chain, a cystine-rich region with 4 conserved disulfide bonds, and an immunoglobulin-like domain. The intracellular domain is also composed of 3 regulatory regions: a membrane proximal domain with a Tyrl003 phosphorylation site, a tyrosine kinase catalytic domain with Tyrl234 and Tyrl235 phosphorylation sites, and a c-terminal multifunctional binding domain with Tyrl349 and Tyrl356 binding tyrosine. After HGF is combined with the extracellular domain of Met, MET phosphorylation is induced, various intercellular factors such as GABl (growth factor receptor binding protein-1), GAB2 (growth factor receptor binding protein-2) and the like are recruited in the C-terminal multifunctional region, and molecules such as SHP2, P13K and the like are further attracted to be combined, so that RAS/MAPK, P13K/AKT, JAK/STAT pathways and the like are activated, and the growth, migration, proliferation and survival of cells are regulated. With the progress of research, MET has been demonstrated to be abnormally expressed or gene amplified in various malignant tumors, which is closely related to the prognosis of tumor patients. The MET gene in many primary tumors shows the phenomenon of gene amplification or high expression, including lung cancer, gastric cancer, colorectal cancer, liver cancer and the like. Among lung cancers, 61% of Non-Small cell lung cancers (NSCLC) and 35% of Small Cell Lung Cancers (SCLC) have MET gene amplification phenomenon, and NSCLC patients with high MET gene copy number have relatively poor prognosis. With the research on MET in tumor aspects being deepened and expanded in recent years, MET gradually becomes an important target of antitumor therapy, particularly, an inhibitor aiming at HGF/MET targeted therapy has good antitumor effect, and the development of MET target inhibitor has great market value.

Target-based therapies are considered to be the direction of development for future cancer treatments. The marketed drugs sorafenib (dojimei) and Lenvatinib (Lenvatinib, E7080) are both multi-kinase inhibitors targeting VEGF receptors and are multi-target kinase inhibitors, but the tumor inhibition effect of the drugs is to be improved.

Disclosure of Invention

In view of the above, it is desirable to provide a substituted urea compound having more prominent tumor-inhibiting effect, a pharmaceutically acceptable salt thereof or a solvate thereof, an application thereof, a medicament thereof, and a pharmaceutical composition.

The invention provides a substituted urea compound, pharmaceutically acceptable salt or solvate thereof, which has a structure shown in a formula (I):

wherein:

R₁selected from methoxy or deuterated methoxy;

R₂selected from methyl, deuterated methyl,

Any one of the structures of (1);

R₃is selected from any one of F, Cl, Br and I.

In one embodiment, R₃Selected from F or Cl.

In one embodiment, the pharmaceutically acceptable salt is a basic salt of an organic or inorganic acid.

In one embodiment, the solvate is a hydrate.

The invention also provides application of the substituted urea compound, the pharmaceutically acceptable salt thereof or the solvate thereof in preparing an inhibitor of one of MET, VEGFR, PDGFR and RET, or in preparing a multi-target inhibitor of two or more of MET, VEGFR, PDGFR and RET.

The invention also provides the use of the substituted urea compounds, pharmaceutically acceptable salts thereof or solvates thereof in the preparation of a medicament or pharmaceutical composition for modulating the activity of a kinase, or treating a disease associated with a kinase, the kinase comprising one or more of MET, VEGFR, PDGFR and RET.

The present invention also provides a medicament or pharmaceutical composition for treating an angiogenesis abnormality disease, comprising the substituted urea compound according to the present invention, a pharmaceutically acceptable salt thereof or a solvate thereof and a physiologically acceptable carrier.

In one embodiment, the angiogenesis abnormality disease includes at least one of cancer, retinal angiogenesis, neovascular glaucoma, inflammatory disease, and diabetic retinopathy.

In one embodiment, the cancer comprises breast cancer, respiratory tract cancer, brain cancer, reproductive organ cancer, digestive tract cancer, urinary tract cancer, eye cancer, liver cancer, skin cancer, head and/or neck cancer, lymphoma, sarcoma, leukemia, thyroid cancer, parathyroid cancer, and/or their distal metastases.

In one embodiment, the cancer is one or more of colorectal cancer, esophageal cancer, gastric cancer, and liver cancer.

The substituted urea compound, the pharmaceutically acceptable salt thereof or the solvate thereof provided by the invention is connected with a substituted urea structure with cyclopropyl and substituted or unsubstituted phenyl through a quinazoline structure which is substituted by dimethoxy, or methoxy and alkoxy containing morpholine heterocycle respectively, and the substituted urea compound can effectively inhibit the enzymatic activity of one or more of MET, VEGFR, PDGFR and RET through experimental verification, and has excellent tumor inhibition effect. The substituted urea compound, the pharmaceutically acceptable salt thereof or the solvate thereof can be used as a kinase inhibitor to prepare a medicament or a pharmaceutical composition for treating diseases related to abnormal angiogenesis.

In particular, due to the heterogeneity of tumors and the complexity of cancer treatment, small molecule drugs with a single target point can only be effective against a small part of tumors, are easy to generate drug resistance, and have poor treatment effect. The multi-target inhibitor can improve the curative effect to a certain extent and prolong the drug resistance generation time, and the multi-target therapy is considered to be the development direction of future cancer treatment. The substituted urea compound, the pharmaceutically acceptable salt thereof or the solvate thereof provided by the invention can be used as a multi-target inhibitor of MET, VEGFR, PDGFR and RET, and has wide application prospect in multi-target therapy of cancer.

Drawings

FIG. 1 is a graph of the concentration-effect relationship of terfenadine on the hERG potassium channel;

FIG. 2 is a graph of the concentration-effect relationship of compound I-1 to the hERG potassium channel for an example of the present invention;

FIG. 3 is a graph of the concentration-effect relationship of compound I-2 on the hERG potassium channel for an example of the present invention;

FIG. 4 is a graph showing the results of the inhibition of tumor volume of KYSE-410 esophageal cancer mice by Compound I-2 according to the example of the present invention;

FIG. 5 is a graph showing the results of body weight changes during administration of Compound I-2 of the present invention to KYSE-410 esophageal cancer mice;

FIG. 6 shows the results of the inhibition of tumor volume in HT-29 colorectal cancer mice by Compound I-2 according to the example of the present invention;

FIG. 7 is a graph showing the results of the inhibition of compound I-2 to the tumor volume of BGC-823 gastric cancer mice;

FIG. 8 is a graph showing the results of the inhibition of tumor volume of SMMC-7721 hepatoma mice by Compound I-2 of the present invention.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention will be described in further detail with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

The embodiment of the invention provides a substituted urea compound, which has the following structural formula:

wherein:

R₁selected from methoxy or deuterated methoxy;

R₂selected from methyl, deuterated methyl,

Any one of the structures of (1);

R₃is selected from any one of F, Cl, Br and I.

The substituted urea compounds described in the embodiments of the present invention include reactive group a and reactive group B. The active group A has a quinazoline ring structure substituted by dimethoxy or methoxy and alkoxy containing morpholine heterocycle respectively. The active group B has a cyclopropyl group and a substituted or unsubstituted phenyl substituted urea structure. The active group A and the active group B are combined through oxygen linkage, and the substituted urea compound has excellent tumor inhibition effect, particularly is used as a multi-target inhibitor for regulating kinase activity, is used for regulating cell activity such as proliferation, differentiation, programmed cell death, migration and chemotaxis, and more particularly can effectively inhibit the enzyme activity of MET, VEGFR, PDGFR and RET, thereby effectively treating diseases related to abnormal angiogenesis, particularly abnormal proliferation diseases accompanied with the angiogenesis.

The present embodiment also provides a compound which undergoes metabolism such as oxidation, reduction, hydrolysis, conjugation, etc. in vivo to exhibit the activity of the substituted urea compound, or a compound which undergoes metabolism such as oxidation, reduction, hydrolysis, etc. in vivo to produce the substituted urea compound, such as a pharmaceutically acceptable salt thereof, and a solvate of the compound, which is also included in the present invention, preferably a hydrate of the compound.

Preferably, the pharmaceutically acceptable salt is a basic salt of an organic acid or an inorganic acid; more preferably a hydrochloride, hydrobromide, hydroiodide, sulphate, phosphate, acetate, trifluoroacetate, thiocyanate, maleate, hydroxymaleate, glutarate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, benzoate, salicylate, phenylacetate, cinnamate, lactate, malonate, pivalate, succinate, fumarate, malate, mandelate, tartrate, gallate, gluconate, laurate, palmitate, pectate, picrate, citrate or a combination thereof.

The substituted urea compound, R₃Is selected from any one of F, Cl, Br and I. In one embodiment, R is₃Preferably F or Cl, and the substituted urea compound may be selected from any one of the following structural formulas:

the substituted urea compounds described in the embodiments of the present invention may be prepared by various methods, and the embodiments of the present invention provide a method for preparing the substituted urea compounds with a high yield, including the steps of:

(1) reacting the compounds (Ia) and (Ib) to obtain a compound (Ic);

(2) reacting the compound (Ic) and the compound (Id) to obtain a compound (Ie);

(3) reacting the compound (Ie) with the compound (If) to obtain the substituted urea compound;

wherein R is₁Selected from methoxy or deuterated methoxy;

R₂selected from methyl, deuterated methyl,

Any one of the structures of (1);

R₃is selected from any one of F, Cl, Br and I.

Compound (Ia) and compound (Ib) are pharmaceutical intermediates, and can be obtained by purchase. Amidating the compound (Ia) and the compound (Ib) in the step (1) under the condition of basic catalysis to obtain a compound (Ic). The used basic catalyst can be organic base or inorganic base, preferably, the basic catalyst is at least one of sodium bicarbonate, sodium carbonate, potassium carbonate and triethylamine; the solvent can be at least one selected from tetrahydrofuran and water mixed solvent, ethanol and water mixed solvent, acetone and water mixed solvent and dichloromethane, wherein in the mixed solvent, the volume ratio of the organic solvent to the water is 1: 10 to 10: 1; the reaction temperature is-10 ℃ to 50 ℃.

Amidation of compound (Ic) and compound (Id) in step (2) under basic catalytic conditions to give compound (Ie). Compound (Id) is cyclopropylamine, and is commercially available. The used basic catalyst can be organic base or inorganic base, preferably, the basic catalyst is at least one of sodium bicarbonate, sodium carbonate, potassium carbonate and triethylamine; the solvent used is preferably at least one of tetrahydrofuran, dichloromethane and acetonitrile; the reaction temperature is 10-70 ℃.

The compound (If) is a pharmaceutical intermediate, and can be obtained by purchase. In the step (3), the compound (Ie) and the compound (If) are etherified under the alkaline catalysis condition to obtain the substituted urea compound, the used catalyst is strong alkali, preferably, the alkaline catalyst is at least one of sodium hydroxide, potassium sodium hydroxide and sodium hydride; the solvent is preferably N, N-dimethylformamide or dimethyl sulfoxide, and the reaction temperature is 30-130 ℃.

The embodiment of the invention also provides application of the substituted urea compound, the pharmaceutically acceptable salt thereof or the solvate thereof in preparing an inhibitor of one of MET, VEGFR, PDGFR and RET, or in preparing a multi-target inhibitor of two or more of MET, VEGFR, PDGFR and PET.

The VEGF includes one or more of VEGFR1, VEGFR2, and VEGFR 3. The PDGFR comprises pdgfra and/or pdgfrp.

The embodiment of the invention also provides application of the substituted urea compound, the pharmaceutically acceptable salt thereof, the solvate thereof or one or more inhibitors of MET, VEGFR, BRAF, PDGFR and/or RET thereof in preparing a medicine or a pharmaceutical composition for regulating kinase activity or treating kinase related diseases. The pharmaceutical composition comprises a drug effect component consisting of the substituted urea compound or the pharmaceutically acceptable salt of the compound or the solvate of the compound and a physiologically acceptable carrier.

The pharmaceutical composition may further comprise other pharmaceutically active agents.

Such other pharmaceutically active agents include, but are not limited to, at least one of the following list: PD-1, PD-L1, lenalidomide, aldesleukin, interferon, amrubicin, arsenic trioxide, 5-azacytidine, capecitabine, carboplatin, custard, simon interleukin, daunorubicin, chlorambucil, cisplatin, cladribine, clodronic acid, cyclophosphamide, cytarabine, floxuridine, fluconazole, fludarabine, 5-fluorodeoxyuridine monophosphate, 5-fluorouracil, gemcitabine, gemtuzumab ozogamicin, imatinib mesylate, idarubicin, ifosfamide, interferon alpha, interferon-alpha 2, interferon alpha-2A, interferon alpha-2B, interferon alpha-n 1, interferon alpha-n 3, interferon beta, interferon gamma-1 a, interleukin-2, intron A, Iressa, Doxol, Doxorubicin, Fluocil, Doxorubicin, Fluocib, Doxorubicin, Fluocil, Doxorubicin, Fluocil, Doxorubicin, Fluben, Lutrazone, Fluben, Lutraben, Fluben, Lumbia, Lutraben, Lumbia, and so, Lutrazone, Ludoxine, Lumbia, and so, Ludoxine, Lumbia, Ludoxine, Lutrazone, Lumbia, Ludoxine, Lutrazone, Ludoxine, and so, Ludoxine, Lutrazone, Ludoxine, Irinotecan, doxorubicin citrate liposome, temozolomide.

The angiogenesis abnormal diseases comprise cancer, neovascular glaucoma, retinal angiogenesis, diabetic retinopathy and inflammatory diseases. The inflammatory diseases include osteoarthritis, rheumatoid arthritis, psoriasis, and delayed hypersensitivity.

Preferably, the abnormal angiogenesis disease is cancer. The cancer includes breast cancer, respiratory tract cancer, brain cancer, cancer of the reproductive organs, cancer of the digestive tract, cancer of the urinary tract, cancer of the eye, liver cancer, skin cancer, cancer of the head and/or neck, lymphoma, sarcoma, leukemia, thyroid cancer, parathyroid cancer and/or their distant metastases. More preferably, the cancer is one or more of colorectal cancer, esophageal cancer and gastric cancer.

Preferably, the breast cancer is invasive ductal carcinoma, invasive lobular carcinoma, ductal carcinoma in situ, or lobular carcinoma in situ; the respiratory cancer is small cell lung cancer, non-small cell lung cancer, bronchial adenoma, or pleuropulmonary blastoma; the brain cancer is brain stem tumor, sub-ocular glioma, cerebellar astrocytoma, brain astrocytoma, medulloblastoma, ependymoma, neuroectodermal or pineal tumor. The genital tumor is prostate cancer, testicular cancer, endometrial cancer, cervical cancer, ovarian cancer, vaginal cancer, vulvar cancer, or uterine sarcoma; the digestive tract cancer is anal cancer, colon cancer, colorectal cancer, esophageal cancer, gallbladder cancer, gastric cancer, pancreatic cancer, rectal cancer, small intestine cancer or salivary gland cancer; the cancer of the urethra is bladder cancer, penile cancer, kidney cancer, renal pelvis cancer, ureter cancer, or cancer of the urethra; the eye cancer is intraocular melanoma or retinoblastoma; the liver cancer is hepatocellular carcinoma, hepatocellular carcinoma with or without fibroplasia change, cholangiocarcinoma or mixed hepatocellular cholangiocarcinoma; the skin cancer is squamous cell carcinoma, Kaposi's sarcoma, malignant melanoma, Mercker's cell skin cancer, or non-melanoma skin cancer; the head and neck cancer is laryngeal cancer, hypopharyngeal cancer, nasopharyngeal cancer, oropharyngeal cancer, lip cancer, or oral cancer; the lymphoma is AIDS-related lymphoma, non-Hodgkin's lymphoma, cutaneous T-cell lymphoma, Hodgkin's disease, or central nervous system lymphoma; the sarcoma is soft tissue sarcoma, osteosarcoma, malignant fibrous histiocytoma, lymphosarcoma, or rhabdomyosarcoma; the leukemia is acute myeloid leukemia, acute lymphocytic leukemia, chronic myelogenous leukemia or hairy cell leukemia.

In general, the use of a cytotoxic inhibitor and/or a cell proliferation inhibitor and/or tumor immunotherapy and/or gene therapy in combination with a substituted urea compound, a pharmaceutically acceptable salt thereof, a solvate thereof, or a composition having the same of the embodiments of the present invention can achieve at least one of the following objectives:

better efficacy in reducing tumor growth or even eliminating tumors than either administration alone;

reducing the amount of monotherapy active agent administered;

are more readily tolerated by patients than monotherapy, have fewer adverse drug complications than therapy with the active agents alone and certain other combination treatments;

is capable of treating a wide variety of different cancer types in mammals, particularly humans;

higher response rates in the treated patients;

longer survival times are achieved in the treated patients compared to standard chemotherapeutic methods;

longer time is required for tumor development;

compared to the antagonistic examples produced by the combination of known anticancer active agents, the efficacy and tolerability results are at least as good as when the active agents are administered alone.

Example 1

The synthetic route for compound I-11- (2-chloro-4- ((6, 7-dimethoxyquinazolin-4-yl) oxy) phenyl) -3-cyclopropylurea is as follows:

the synthesis method comprises the following steps:

(1) 4-amino-2-chloro-phenol (12.7g,0.1mol) and sodium hydrogencarbonate (16.8g, 0.2mol) were added to tetrahydrofuran (100ml) and water (100ml), and stirred at room temperature to be in a uniform suspension state (solution A). Phenyl chloroformate (18.8g, 0.12mol) was dissolved in tetrahydrofuran (50mL) to form a solution (solution B). And (3) dropwise adding the solution B into the solution A, stirring at room temperature for reaction after the dropwise adding is finished, tracking the reaction by TLC, analyzing and confirming the completion of the reaction, and then carrying out post-treatment. The post-treatment comprises the following steps: and (2) standing and separating the reaction solution, extracting the water phase by using 100mL ethyl acetate, washing the organic phase by using saturated saline solution for 3 times, adding anhydrous sodium sulfate for drying, carrying out suction filtration, carrying out vacuum concentration on the filtrate to obtain a solid, adding ethyl acetate (50mL), heating, refluxing and dissolving the solid, adding petroleum ether (100mL), cooling to room temperature, stirring and crystallizing for 5 hours, carrying out suction filtration on the separated crystal, and drying to obtain 15.3g of a compound (I-1c), wherein the yield is 62%.

(2) Compound (I-1c) (12.4g, 0.05mol), cyclopropylamine (5.7g, 0.1mol) and triethylamine (5g, 0.05mol) were added to acetonitrile (150mL), and the reaction was stirred with heating at 50 ℃ and followed by TLC, and after completion of the reaction was confirmed by analysis, the reaction was worked up. The post-treatment comprises the following steps: cooling to room temperature, filtering, washing a filter cake with a small amount of acetonitrile, and vacuum-drying the obtained solid at 45 ℃ to obtain 9.03g of the compound (I-1e), wherein the yield is 86%.

(3) Compound (I-1e) (5.88g, 0.028mol), 4-chloro-6, 7-dimethoxyquinazoline (4.49g, 0.02mol) and sodium hydroxide (0.96g, 0.24mol) were added to DMF (50mL), and the reaction was stirred with heating at 50 ℃ and followed by TLC, and after completion of the reaction was confirmed by analysis, post-treatment was carried out. The post-treatment comprises the following steps: 100mL of water was added to the reaction solution, crystallization was carried out at room temperature with stirring, suction filtration was carried out, and the filter cake was washed with purified water 2 times. The obtained solid was slurried in ethanol (250mL), filtered under suction, and dried under vacuum to give 7.13g of compound (I-1) in 86% yield.

The nuclear magnetic data of the obtained compound (I-1) were: 1H NMR (400MHz, DMSO). delta. (ppm): 8.61(s,1H),8.25(d,1H),8.00(s,1H),7.59(s,1H),7.54(d,1H),7.43(s,1H),7.30(dd,1H),7.22(d,1H),4.11-3.96(m,6H),2.69-2.60(m,1H),0.72(q,2H),0.56-0.40(m, 2H); ESI-MS (m/z): 415.2[ M +1 ].

Example 2

The synthetic route for compound I-21- (2-fluoro-4- ((6, 7-dimethoxyquinazolin-4-yl) oxy) phenyl) -3-cyclopropylurea is as follows:

the synthesis method can refer to example 1, the equivalent ratio and the reaction conditions in step (1) and step (2) are not changed, the equivalent of the reactants in step (3) is not changed, and the reaction conditions are heating and stirring reaction at 40 ℃. The post-treatment comprises the following steps: 100mL of water was added to the reaction solution, crystallization was carried out at room temperature with stirring, suction filtration was carried out, and the filter cake was washed with purified water 2 times. The obtained solid was slurried in ethanol (200mL), filtered, and vacuum dried to give 6.05g of compound (I-2) in 76% yield.

The nuclear magnetic data of the obtained compound (I-2) were: 1H NMR (400MHz, DMSO). delta. (ppm): 8.62(s,1H),8.26(d,1H),8.03(s,1H),7.62(s,1H),7.55(d,1H),7.43(s,1H),7.32(d,1H),7.20(d,1H),4.12-3.96(m,6H),2.70-2.60(m,1H),0.73(q,2H),0.57-0.41(m, 2H); ESI-MS (M/z):399[ M +1 ].

Example 3

The synthetic route for compound I-31- (2-chloro-4- ((6-methoxy-7- (3-morpholinopropoxy) quinazolin-4-yl) oxy) phenyl) -3-cyclopropylurea is as follows:

the synthesis method can refer to example 1, the equivalent ratio and the reaction conditions in step (1) and step (2) are not changed, the equivalent of the reactants in step (3) is not changed, and the reaction conditions are heating and stirring reaction at 50 ℃. The post-treatment comprises the following steps: the reaction solution was concentrated under reduced pressure to remove about 40mL of DMF solvent, 50mL of water was added to the reaction solution, followed by crystallization at room temperature with stirring, suction filtration, and washing of the filter cake with purified water 2 times. The resulting solid was recrystallized from ethanol (100mL), filtered under suction, and dried under vacuum at 45 ℃ to give 4.86g of Compound (I-3) in 46% yield.

The nuclear magnetic data of the compound (I-3) obtained were:¹H NMR(400MHz,DMSO)δ(ppm)：8.61(s,1H),8.25(d,1H),8.00(s,1H),7.59(s,1H),7.54(d,1H),7.43(s,1H),7.30(d,2.5Hz,1H),7.22(d,1H),4.31(t,2H),4.08(s,3H),3.71-3.77(m,4H),2.60-2.65(m,2H),2.44-2.57(m,5H),2.07-2.18(m,2H),0.72(q,J＝6.5Hz,2H),0.56–0.40(m,2H)；ESI-MS(m/z):528.2[M+1]。

example 4

The synthetic route for compound I-41- (2-fluoro-4- ((6-methoxy-7- (3-morpholinopropoxy) quinazolin-4-yl) oxy) phenyl) -3-cyclopropylurea is as follows:

the synthesis method can refer to example 1, the equivalent ratio and the reaction conditions in step (1) and step (2) are not changed, the equivalent of the reactants in step (3) is not changed, and the reaction conditions are heating and stirring reaction at 70 ℃. The post-treatment comprises the following steps: the reaction solution was concentrated under reduced pressure to remove about 40mL of DMF solvent, 50mL of water was added to the reaction solution, followed by crystallization at room temperature with stirring, suction filtration, and washing of the filter cake with purified water 2 times. The obtained solid was recrystallized from ethanol (150mL), filtered under suction, and vacuum-dried at 45 ℃ to give 7.36g of compound (I-4) with a yield of 72%.

The nuclear magnetic data of the obtained compound (I-4) were:¹H NMR(400MHz,DMSO)δ(ppm)：8.56(s,1H),8.12-8.19(m,2H),7.54(d,1H),7.39(d,1H),7.30-7.33(m,1H),7.07-7.09(m,1H),6.80(m,1H),4.25(t,2H),3.99(s,3H),3.64-3.70(m,4H),2.53-2.58(m,2H),2.39-2.50(m,5H),2.05-2.16(m,2H),0.65(m,2H),0.42(m,2H)；ESI-MS(m/z):512.2[M+1]。

example 5

The synthetic route for compound I-51- (2-chloro-4- ((6-methoxy-7- (2-morpholinoethoxy) quinazolin-4-yl) oxy) phenyl) -3-cyclopropylurea is as follows:

the synthesis method and the post-treatment were carried out in accordance with example 3 to obtain compound (I-5) with a yield of 46%.

The nuclear magnetic data of the obtained compound (I-5) were:¹H NMR(400MHz,DMSO)δ(ppm)：8.60(s,1H),8.24(d,1H),8.01(s,1H),7.61(s,1H),7.55(d,1H),7.42(s,1H),7.31(m,1H),7.21(d,1H),4.25-4.32(m,2H),4.11-3.96(m,6H),3.66-3.73(m,4H),2.85-2.94(m,2H),2.69-2.54(m,5H),0.72(q,2H),0.56–0.40(m,2H)；ESI-MS(m/z):514.2[M+1]。

example 6

The synthetic route for compound I-61- (2-fluoro-4- ((6-methoxy-7- (2-morpholinopropoxy) quinazolin-4-yl) oxy) phenyl) -3-cyclopropylurea is as follows:

the synthesis method and the post-treatment were carried out in accordance with example 4 to obtain compound (I-6) with a yield of 72%.

The nuclear magnetic data of the obtained compound (I-6) were:¹H NMR(400MHz,DMSO)δ(ppm)：8.57(s,1H),8.14-8.19(m,2H),7.56(d,1H),7.41(d,1H),7.31-7.35(m,1H),7.08-7.10(m,1H),6.82(m,1H),4.28-4.35(m,2H),3.97-3.99(m,6H),3.69-3.77(m,4H),2.88-2.98(m,2H),2.73–2.57(m,5H),0.65(m,2H),0.42(m,2H)；ESI-MS(m/z):498.2[M+1]。

experimental example 1 in vitro inhibition of enzyme Activity of substituted Urea Compounds I-1 to I-6

Z' -LYTE from Thermo Fisher scientific was used^TMThe inhibitory effects of the substituted urea compounds I-1 to I-6 on the activity of various enzymes were examined. The kinases used in the assay were all commercially available and the assay was performed according to Z' -LYTE^TMThe method is carried out by a conventional operation method.

The reagent and the preparation condition are as follows:

1. 1 Xkinase buffer A (50mM HEPES (pH7.5), 0.01% BRIJ-35,10mM MgCl₂,4mM MnCl₂,1mM EGTA,2mM DTT)

1 Xkinase buffer B (50mM HEPES (pH7.5), 0.01% BRIJ-35,10mM MgCl₂,1mM EGTA)

In the kinase reaction, the ATP solution, substrate, enzyme and compound are diluted with 1 Xkinase buffer A or 1 Xkinase buffer B.

2. Working solution of test compound

Accurately weighing the compounds obtained in the examples 1-6, adding a dimethyl sulfoxide (DMSO) solvent to form a mother solution, and then preparing the solution of the compounds I-1 to I-6 to be detected to the required concentration by using a kinase buffer solution;

3. 4 x ATP (adenosine triphosphate) working solution

ATP was prepared to 4-fold the final concentration of the reaction using 1 Xkinase buffer B.

4、2×Z’-LYTE^TMPeptide substrate/enzyme working solutions

VEGFR1 enzyme/peptide substrate mix:

peptide substrates Tyr 04 and VEGFR1 enzymes were formulated to 2-fold the final concentration of the reaction with 1 × kinase buffer a. 10 μ L of VEGFR1 enzyme/peptide substrate mixture for the final kinase reaction included 2.64ng-12.5ng VEGFR1 and 2 μ M Tyr 04, 50mM HEPES (pH7.5), 0.01% BRIJ-35,10mM MgCl₂,2mM MnCl₂,1mM EGTA,1mM DTT。

VEGFR3 enzyme/peptide substrate mix:

peptide substrates Tyr 04 and VEGFR3 enzymes were formulated to 2-fold the final concentration of the reaction with 1 × kinase buffer a. 10 μ L of VEGFR3 enzyme/peptide substrate mixture for the final kinase reaction included 2ng-20ng VEGFR3, 2 μ M Tyr 04, and 50mM HEPES (pH7.5), 0.01% BRIJ-35,10mM MgCl₂，2mM MnCl₂,1mM EGTA,1mM DTT。

VEGFR2 enzyme/peptide substrate mix:

peptide substrates Tyr 01 and VEGFR2 enzymes were formulated with 1 × kinase buffer B to 2-fold the final concentration of the reaction. 10 μ L of VEGFR2 enzyme/peptide substrate mixture for the final kinase reaction included 0.75ng-15ng VEGFR2, 2 μ M Tyr 01 and 50mM HEPES (pH7.5), 0.01% BRIJ-35,10mM MgCl₂,1mM EGTA。

PDGFR α enzyme/peptide substrate mixture:

peptide substrates Tyr 04 and PDGFR α enzymes were formulated with 1 × kinase buffer a to 2-fold the final concentration of the reaction. mu.L of PDGFR alpha enzyme/peptide substrate mixture at the final kinase reaction contains 1.54ng-22.6ng PDGFR alpha, 2. mu.M Tyr 04, 50mM HEPES (pH7.5), 0.01% BRIJ-35,10mM MgCl₂,2mM MnCl₂，1mM EGTA,1mM DTT。

PDGFR β enzyme/peptide substrate mixture:

peptide substrates Tyr 04 and PDGFR β enzyme were formulated with 1 × kinase buffer a to 2-fold the final concentration of the reaction. Final kinase reaction 1mu.L of PDGFR beta enzyme/peptide substrate mixture comprising 7ng-50ng PDGFR beta, 2. mu.M Tyr 04, 50mM HEPES (pH7.5), 0.01% BRIJ-35,10mM MgCl₂，2mM MnCl₂,1mM EGTA,1mM DTT。

RET enzyme/peptide substrate mixture:

peptide substrates Tyr02 and RET enzymes were formulated to 2-fold the final concentration of the reaction with 1 × kinase buffer B. Final kinase reaction 10. mu.L of RET enzyme/peptide substrate mixture comprised 0.49ng-3.64ng RET, 2. mu.M Tyr02 and 50mM HEPES (pH7.5), 0.01% BRIJ-35,10mM MgCl₂,1mM EGTA。

MET enzyme/peptide substrate mixture:

peptide substrates Tyr 06 and MET enzymes were formulated to 2-fold the final concentration of the reaction with 1 × kinase buffer B. The METase/peptide substrate mixture for the final kinase reaction included 0.49ng-7.84ng MET, 2. mu.M Tyr 06 and 50mM HEPES (pH7.5), 0.01% BRIJ-35,10mM MgCl₂,1mM EGTA。

Wherein HEPES is N- (2-hydroxyethyl) piperazine-N' -2-ethanesulfonic acid, BRIJ-35 is dodecyl polyethylene glycol ether, EGTA is ethylene glycol bis (2-aminoethyl ether) tetraacetic acid, Tyr is tyrosine, Ser is serine, DTT is dithiothreitol, MEK1 is mitogen-activated protein kinase-1, ERK2 is extracellular regulated protein kinase-2, and Thr is threonine.

(di) Z' -LYTE^TMThe screening protocol and assay conditions were:

adding a compound to be detected diluted by a buffer solution, an enzyme/peptide substrate mixed solution and ATP (adenosine triphosphate) into a 384-well plate, shaking the plate for 30 seconds, and culturing the plate for 1 hour at room temperature to perform a kinase reaction;

adding a fluorescence enhancer into each hole, and incubating for 1 hour at room temperature;

respectively reading the fluorescence intensity data of 445nm and 520nm of each hole by using a fluorescence analyzer, and processing the data to obtain the inhibition rate IC of the substituted urea compound on the activities of VEGFR1, VEGFR2, VEGFR3, PDGFR alpha, PDGFR beta, RET and MET₅₀The value is obtained.

Specifically, the data processing method comprises the following steps: the data read by the fluorescence analyzer are collated, and the Ratio of the fluorescence intensity at 445nm and 520nm of each well (Ratio 445/520) is calculated according to a formula) And the relative inhibition of each well. The inhibition ratio IC₅₀Values are the relative inhibition measured after dilution of the concentration of the active sample containing the compound, calculated by Xlfit software mapping.

The results of the experiment are shown in table 1:

TABLE 1 inhibition of enzyme Activity by Compounds I-1 to I-6

From the above experimental results, it can be seen that the half inhibitory concentrations IC of the substituted urea compounds I-1 to I-6 provided in the examples of the present invention against VEGFR1, VEGFR2, VEGFR3, PDGFR α, PDGFR β, RET and MET₅₀The values are all in nanomolar level, which shows that the substituted urea compounds I-1 to I-6 have stronger binding effect on each target spot of VEGFR1, VEGFR2, VEGFR3, PDGFR alpha, PDGFR beta, RET and MET, and can effectively inhibit the enzyme activity at extremely low concentration.

EXAMPLE 2 inhibition of hERG (Potassium channel) by substituted Urea Compounds I-1 and I-2

The experimental steps are as follows:

1. cell preparation

HEK-293 cells (from military medical sciences) were washed with PBS (phosphate buffered saline), digested with Tryple (pancreatic substitute) solution, resuspended in DMEM medium (Dulbecco's modified eagle medium) and stored in a centrifuge tube until needed. Before the patch clamp records, the cells are dripped into a perfusion groove on the glass bottom or a culture dish of 35mm, so that the cells have certain density and are in a single separation state.

2. Electrophysiological test

Recording hERG current by adopting a whole-cell patch clamp technology, adding the prepared cell suspension into a culture dish, placing the culture dish on an inverted microscope objective table, and perfusing with extracellular fluid after the cells adhere to the wall;

the glass microelectrode is drawn by a microelectrode drawing instrument in two steps, and the water inlet resistance value of the glass microelectrode is 2-5 MOmega after filling the liquid in the electrode; after establishing a whole-cell recording mode, keeping the clamp potential at-80 mV, giving a depolarization voltage to +60mV with the time course of 850ms, then repolarizing to-50 mV with the time course of 1275ms, and leading out the hERG tail current. The pulse program was repeated every 15 seconds throughout the entire experiment;

after the current is stabilized, extracellular continuous perfusion administration from low concentration to high concentration is adopted. Starting from low concentration, the perfusion is continued until the drug effect is stable, and then the perfusion of the next concentration is performed. The substituted urea compounds I-1 and I-2 prepared in examples 1 and 2 of the present invention were used as test samples, respectively, the test samples were dissolved in DMSO, diluted with extracellular fluid to the desired concentration, and tested for blocking effect of 5 test samples of different concentrations and 4 positive controls of concentration on hERG tail current, the positive control being Terfenadine (Terfenadine), which has toxic and side effects on heart.

(II) data acquisition and analysis

Stimulation issuing and signal acquisition are carried out through PatchMadter software; the patch clamp amplifier amplifies the signal and the filtering is 10 KHz. Further data analysis and curve fitting were performed using FitMaster, EXCEL and SPASS 21.0 et al. Data are presented as mean ± standard deviation. In data processing, the peak value of the tail current and its baseline are corrected when the blocking effect on hERG is judged. The effect of each compound at different concentrations is expressed as the inhibition of the wake. Inhibition rate is 100 × (tail current peak before dosing-tail current peak after dosing)/tail current peak before dosing. Taking the concentration of the test substance as a horizontal axis and the current inhibition rate of the normalization treatment as a vertical axis, making a concentration-effect curve, and fitting by a Hill equation to obtain the semi-inhibitory concentration IC of the test substance₅₀Numerical values.

(III) results of the experiment

The experimental results are shown in FIGS. 1-3 and Table 2, and the IC of the substituted urea compounds I-1 and I-2 provided in the examples of the present invention on hERG current₅₀Both values were greater than 30. mu.M and showed no significant inhibition of the hERG channel, indicating that compounds I-1 and I-2 provided in the examples of the invention are not cardiotoxic.

TABLE 2 IC of Compounds I-1, I-2 on hERG Current₅₀Value of

Compound or positive control	IC₅₀(μM)	Completion sample size (N)
			I-1	>30	2
I-2	>30	2
			Terfenadine (Positive control)	0.042	3

An animal experiment is adopted to detect the in-vivo anti-tumor activity of the substituted urea compound I-2, and the anti-tumor activity is compared with the cabozantinib and the lenvatinib, wherein the tumors are esophageal cancer, colorectal cancer tumor, gastric cancer and liver cancer.

EXAMPLE 3 anti-esophageal cancer tumor Activity of substituted Urea Compound I-2 in vivo

1. Establishment of tumor model

KYSE-410 esophageal cancer cells (from military medical academy of sciences) were treated with 10% fetal calf serum-containing high-glucose DMEM at 37 deg.C and 5% CO₂Performing conventional culture in an incubator, after the cells are propagated for three generations in vitro, digesting and collecting the cells when the cells grow to more than 80% of fusion rate and reach the required amount, and suspending the cells with matrigel at a ratio of 1: 1. Will be about 2X 10⁶Gastric cancer cells, esophageal cancer cells and colorectal cancer cells were injected into the left axilla of each nude mouse, respectively.

2. Grouping and administration of laboratory animals

When the tumor grows to 100mm³～300mm³Thereafter, animals were randomly grouped into groups of 6 animals each, and fed with different administration forms, respectively:

(1) model group: gavage 0.5% sodium carboxymethylcellulose solvent daily;

(2) compound I-2 test group 1: gavage 3mg/kg (mouse body weight) of compound I-2 solution daily;

(3) compound I-2 test group 2: gavage 5mg/kg (mouse body weight) of compound I-2 solution daily;

(4) compound I-2 test group 3: gavage 10mg/kg (mouse body weight) of compound I-2 solution per day;

(5) compound I-2 test group 4: gavage 20mg/kg (mouse body weight) of compound I-2 solution per day;

(6) positive control group 1: gavagant solution 10mg/kg (mouse body weight) per day;

(7) positive control group 2: gavagant solution at 30mg/kg (mouse body weight) was gavaged daily.

The mice were weighed once every same time, the body weight, tumor volume of the mice were recorded, and the Relative Tumor Volume (RTV) was calculated. Wherein, the RTV calculation formula is that RTV is equal to V_t/V₀In which V is_tIs a representation of the tumor volume at day t after administration, V₀Is the tumor volume on the day of administration.

The experimental results are shown in figures 4 and 5, and it can be seen from figure 4 that the inhibition effect of the compound I-2 on esophageal cancer is stronger than that of the positive drug lenvatinib (10mg/kg and 30mg/kg) in large dose from low dose (3mg/kg) to high dose (20mg/kg), which indicates that the anti-esophageal cancer tumor effect of the compound I-2 is better than that of lenvatinib; FIG. 5 shows that the body weight of the mice in the experimental group administered with Compound I-2 remained stable, while the body weight of the mice in the experimental group administered with lenvatinib decreased significantly, indicating that Compound I-2 is more safe and has less side effects than lenvatinib.

Experimental example 4 anti-colorectal cancer tumor Activity of substituted Urea Compound I-2 in vivo

1. Establishment of tumor model

HT-29 colorectal cancer cells (from military medical academy of sciences) were treated with 10% fetal bovine serum in high-glucose DMEM at 37 deg.C and 5% CO₂Performing conventional culture in an incubator, after the cells are propagated for three generations in vitro, digesting and collecting the cells when the cells grow to more than 80% of fusion rate and reach the required amount, and suspending the cells with matrigel at a ratio of 1: 1. Will be about 2X 10⁶Gastric cancer cells, esophageal cancer cells and colorectal cancer cells were injected into the left axilla of each nude mouse, respectively.

2. Grouping and administration of laboratory animals

(1) model group: gavage 0.5% sodium carboxymethylcellulose solvent daily;

(2) compound I-2 test group 1: gavage 2mg/kg (mouse body weight) of compound I-2 solution daily;

(5) positive control group: gavagant solution 10mg/kg (mouse body weight) was gavagant daily.

The experimental results are shown in FIG. 6, and it can be seen from FIG. 6 that the inhibitory effect of compound I-2 on colorectal cancer is stronger than that of the positive drug lenvatinib (10mg/kg) in large dose from low dose (2mg/kg) to high dose (10mg/kg), which indicates that the anti-colorectal cancer tumor effect of compound I-2 is better than that of lenvatinib.

Experimental example 5 anti-gastric cancer tumor Activity of substituted Urea Compound I-2 in vivo

1. Establishment of tumor model

BGC-823 gastric cancer cells (from military medical academy of sciences) were cultured in 10% fetal bovine serum-containing high-glucose DMEM at 37 deg.C and 5% CO₂Performing conventional culture in an incubator, after the cells are propagated for three generations in vitro, digesting and collecting the cells when the cells grow to more than 80% of fusion rate and reach the required amount, and suspending the cells with matrigel at a ratio of 1: 1. Will be about 2X 10⁶Gastric cancer cells, esophageal cancer cells and colorectal cancer cells were injected into the left axilla of each nude mouse, respectively.

2. Grouping and administration of laboratory animals

(1) model group: gavage 0.5% sodium carboxymethylcellulose solvent daily;

(5) positive control group: gavage 30mg/kg (mouse body weight) of cabozantinib solution daily.

The experimental results are shown in FIG. 7, and it can be seen from FIG. 7 that the gastric cancer inhibitory effect of the compound I-2 is stronger than that of the positive drug cabozantinib (30mg/kg) in large dose from low dose (3mg/kg) to high dose (10mg/kg), which indicates that the anticancer effect of the compound I-2 is better than that of cabozantinib.

Example 6 anti-hepatoma tumor Activity of substituted Urea Compound I-2 in vivo

1. Establishment of tumor model

The SMMC-7721 liver cancer cell (from Jun)Institute of medical sciences) with 10% fetal bovine serum in high-glucose DMEM at 37 ℃ with 5% CO₂Performing conventional culture in an incubator, after the cells are propagated for three generations in vitro, digesting and collecting the cells when the cells grow to more than 80% of fusion rate and reach the required amount, and suspending the cells with matrigel at a ratio of 1: 1. Will be about 2X 10⁶Gastric cancer cells, esophageal cancer cells and colorectal cancer cells were injected into the left axilla of each nude mouse, respectively.

2. Grouping and administration of laboratory animals

(1) model group: gavage 0.5% sodium carboxymethylcellulose solvent daily;

(2) compound I-2 test group 1: gavage 5mg/kg (mouse body weight) of compound I-2 solution daily;

(3) compound I-2 test group 2: gavage 10mg/kg (mouse body weight) of compound I-2 solution per day;

(4) positive control group: gavagant solution 10mg/kg (mouse body weight) was gavagant daily.

The experimental results are shown in FIG. 8, and it can be seen from FIG. 8 that the compound I-2 has strong inhibitory effect on gastric cancer from low dose (3mg/kg) to high dose (10mg/kg), and the compound I-2 has stronger in vivo anti-hepatoma tumor effect than the positive drug lenvatinib at high dose of 10 mg/kg.

The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.

The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the present invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.

Claims

1. A substituted urea compound, a pharmaceutically acceptable salt thereof, having the structure shown in formula (I):

wherein:

R₁selected from methoxy;

R₂is selected from methyl;

R₃is selected from any one of F, Cl, Br and I.

2. The substituted urea compound, pharmaceutically acceptable salt thereof, according to claim 1, wherein R is₃Selected from F or Cl.

3. The pharmaceutically acceptable salt of a substituted urea compound according to claim 1 wherein the pharmaceutically acceptable salt is a basic salt of an organic or inorganic acid.

4. Use of a substituted urea compound according to any one of claims 1 to 3, a pharmaceutically acceptable salt thereof, for the preparation of an inhibitor of one of MET, PDGFR and RET, or for the preparation of a multi-target inhibitor of two or more of MET, VEGFR, PDGFR and RET.

5. Use of a substituted urea compound according to any one of claims 1 to 3, a pharmaceutically acceptable salt thereof, for the manufacture of a medicament or pharmaceutical composition for modulating kinase activity or treating a kinase-associated disease, wherein the kinase is two or more of MET, VEGFR, PDGFR and RET.

6. A medicament or pharmaceutical composition for the treatment of an angiogenic disorder comprising a substituted urea compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3 and a physiologically acceptable carrier.

7. The medicament or pharmaceutical composition of claim 6, wherein the abnormal angiogenesis disease is at least one of cancer, retinal angiogenesis, neovascular glaucoma, inflammatory diseases, and diabetic retinopathy.

8. The medicament or pharmaceutical composition according to claim 7, wherein said cancer is breast cancer, respiratory cancer, brain cancer, cancer of the reproductive organs, cancer of the digestive tract, cancer of the urinary tract, eye cancer, liver cancer, skin cancer, cancer of the head and/or neck, lymphoma, sarcoma, leukemia, thyroid cancer, parathyroid cancer and/or their distant metastases.

9. The medicament or pharmaceutical composition of claim 7, wherein the cancer is one or more of colorectal cancer, esophageal cancer, gastric cancer, and liver cancer.