New uses for quinoline compounds

Abstract

The compound of formula (I) or a pharma- ceutically acceptable salt, solvate, ester, acid, metabolite or prodrug thereof (Y, A, n, R 1 , R 2 and R 3 (wherein M2 phenotype TAMs are defined herein.) By converting M2 phenotype TAMs to M1 phenotype TAMs in the tumor immune microenvironment, the tumor microenvironment is improved, promoting the phagocytic effect of macrophages on tumor cells, thereby further inhibiting tumor growth. [Formula 1] TIFF2025507088000084.tif37147

Description

[Technical field]

[0001] The present invention relates to the biomedical field, in particular to the use of quinoline compounds in anti-cancer drugs. [Background technology]

[0002] Cancer immunotherapy is the new most promising treatment after surgery, chemoradiotherapy and targeted therapy, and is also the focus and hotspot in the current tumor research field. However, the majority of patients in clinical cases still do not respond to immunotherapy, which has been proven to be inseparable from the immune resistance mediated by the tumor microenvironment (TME). Targeting the TME to alleviate immune suppression can restore and reconstruct the body's normal anti-tumor immune defense ability, and enhance the efficacy of immunotherapy.

[0003] Tumor-associated macrophages (TAMs) are an integral part of the TME and play a key role in tumor initiation, development, invasion, and metastasis. Macrophages can be divided into M1 (classically activated macrophages) and M2 (alternatively activated macrophages) according to their phenotype and secreted cytokines.

[0004] M1 macrophages mainly secrete inflammatory cytokines and chemokines, which enhance antigen presentation, promote T cell proliferation and activation, and participate in inflammatory responses, thereby improving immune killing ability against tumor cells. In contrast, M2 macrophages have poor antigen presentation ability, downregulate immune responses, and can inhibit CD8+T cytotoxicity in the TME, which may promote tumor initiation and development. M1 macrophages can be distinguished by indicators such as membrane protein CD80 / 86 and cytokines IL-1β and IL-6 secreted by M1 macrophages. Meanwhile, M2 macrophages can be distinguished by indicators such as the expression levels of Arg-1 and CD206 / 163.

[0005] Numerous studies have found that TAMs in tumor tissues are predominantly M2, which is a key factor in the formation of immunosuppressive TME. Moreover, some studies have shown that certain stimuli can directionally induce the conversion of M2 TAMs to M1 TAMs, thereby inhibiting tumor growth and enhancing the efficacy of cancer immunotherapy. Therefore, targeting TAMs and promoting the polarization of TAMs to M1, as well as utilizing regulators that can promote the conversion of TAMs from M2 to M1, will provide new ideas for future tumor immunotherapy. This kind of agent can significantly inhibit tumor growth in clinical settings, and some solid tumors may even disappear completely after treatment. Summary of the Invention [Problem to be solved by the invention]

[0006] The object of the present invention is to provide compounds that induce the conversion of M2 TAM to M1 TAM, thereby inhibiting tumor growth and improving the efficacy of cancer immunotherapy, which can be used to treat cancers or tumors in which M2 macrophages predominate in the tumor microenvironment. Furthermore, the object of the present invention is to provide compounds that can inhibit the activity of CSF1R kinase and the associated signaling pathway, thereby activating autoimmunity for the treatment of cancer by inhibiting the phosphorylation of CSF1R. [Means for solving the problem]

[0007] Thus, the present invention provides the use of a compound of formula (I), or a pharma- ceutically acceptable salt, solvate, ester, acid, metabolite or prodrug thereof, in the manufacture of a medicament for treating a cancer or tumor in which M2 macrophages predominate in the tumor microenvironment: [ka] (In the formula, Y is [ka] is selected from A is selected from aryl or 6-membered heterocyclyl; n is an integer selected from 1 to 3; R 1 is hydrogen, halo, C 1~6 Alkyl, C 1~6 Haloalkyl, C 1~6 selected from alkoxy and cyano; R 2 and R 3 each independently represents hydrogen, halo, C 1~6 Alkyl, C 1~6 Haloalkyl, C 1~6 alkoxy or R 2 and R 3 together form a phenyl or a 5-membered heterocyclyl).

[0008] In a preferred embodiment, Y is [ka] When Y is [ka] In another embodiment, A is preferably selected from phenyl, N-morpholinyl, N-piperidyl or N-piperazinyl. 1 is selected from hydrogen, fluoro, chloro, methyl, ethyl, propyl, or cyano; R 2 and R 3 are each independently selected from hydrogen, fluoro, chloro, methyl, ethyl, propyl, trifluoromethyl, difluoromethyl, trifluoroethyl, methoxy, ethoxy, propoxy, or R 2 and R 3 combine to form a phenyl or dioxolane.

[0009] In a preferred embodiment, the present invention provides the use of a compound of formula (Ia), or a pharma- ceutically acceptable salt, solvate, ester, acid, metabolite or prodrug thereof, in the manufacture of a medicament for treating a cancer or tumor in which M2 macrophages predominate in the tumor microenvironment: [ka] (In the formula, R 1 , R 2 and R 3 is defined above).

[0010] More preferably, in the formula, R 1 is selected from hydrogen, fluoro, chloro, and methyl; R 2 and R 3 are each independently selected from hydrogen, fluoro, chloro, methyl, and trifluoromethyl, or R 2 and R 3 taken together form a phenyl.

[0011] In a further preferred embodiment, the present invention provides the use of a compound of formula (Ib), or a pharma- ceutically acceptable salt, solvate, ester, acid, metabolite or prodrug thereof, in the manufacture of a medicament for treating a cancer or tumor in which M2 macrophages predominate in the tumor microenvironment: [ka] (In the formula, A, R 1 , R 2 and R 3 is defined above).

[0012] More preferably, A is selected from phenyl or N-morpholinyl, and R 1 is hydrogen, and R 2 and R 3 is each independently selected from hydrogen, fluoro, chloro, methyl, and trifluoromethyl.

[0013] The present invention relates to any one of the compounds of formula (I), formula (Ia) and formula (Ib), or its pharmaceutically acceptable salt, solvate, ester, acid, metabolite or prodrug, which can convert macrophages in the TME from M2, which promotes tumor growth, to M1, thereby changing the TME and achieving the purpose of treating cancer. Furthermore, the compounds of formula (I), formula (Ia) and formula (Ib) of the present invention, or its pharmaceutically acceptable salt, solvate, ester, acid, metabolite or prodrug, can inhibit the activity of CSF1R kinase and related signaling pathways, thereby achieving the purpose of treating cancer.

[0014] The present invention relates to any one of the compounds of formula (I), formula (Ia) and formula (Ib), or a pharma-ceutically acceptable salt, solvate, ester, acid, metabolite or prodrug thereof, for use as a medicament for treating cancers or tumors in which M2 macrophages predominate in the tumor microenvironment.

[0015] The cancer or tumor is selected from one or more of ovarian cancer, cervical cancer, membranous adenocarcinoma, prostate cancer, bladder cancer, lung cancer, thyroid cancer, breast cancer, pancreatic cancer, kidney cancer, stomach cancer, liver cancer, cervical cancer, endometrial cancer, colorectal cancer, nasopharyngeal cancer, esophageal cancer, bile duct cancer, bone metastasis cancer, papillary thyroid cancer, non-small cell lung cancer, small cell lung cancer, colon cancer, solid tumor, brain tumor, lymphoma, glioma, glioblastoma, melanoma, mesothelioma, glioblastoma, osteosarcoma, gastrointestinal stromal tumor, multiple myeloma, bile duct carcinoma sarcoma, and leukemia. Among these, lymphoma includes non-Hodgkin's lymphoma, B-cell or T-cell lymphoma, and leukemia includes chronic myelogenous leukemia or acute myelogenous leukemia.

[0016] The present invention also relates to a method of treating a cancer or tumor in which M2 macrophages predominate in the tumor microenvironment, the method comprising administering to a patient any one of the compounds of formula (I), formula (Ia) and formula (Ib) of the present invention, or a pharma- ceutically acceptable salt, solvate, ester, acid, metabolite or prodrug thereof. [Brief description of the drawings]

[0017] [Figure 1] FIG. 1 shows the effect of compound 2, BLZ945 and PLX3397 on IL-10 secretion after polarization of RAW264.7 cells. [Diagram 2] FIG. 1 shows the effect of compound 2, BLZ945 and PLX3397 on the expression of the molecule CD86 after polarization of RAW264.7 cells. [Diagram 3] FIG. 1 shows the effect of Compound 2 and PLX3397 on mouse body weight after administration to an MC38 cell tumor-bearing mouse model. [Figure 4] FIG. 1 shows the tumor inhibitory effects of Compound 2 and PLX3397 after administration to a MC38 cell tumor-implanted mouse model. [Diagram 5] FIG. 1 shows the effects of Compound 2 and PLX3397 on the proportion of M1 macrophages in the tumor microenvironment after administration to a mouse model with an MC38 cell tumor xenograft. [Figure 6] FIG. 1 shows the effect of Compound 2 and PLX3397 on mouse body weight after administration to a 4T1 cell tumor-implanted mouse model. [Figure 7] FIG. 1 shows the tumor inhibitory effects of Compound 2 and PLX3397 after administration to a 4T1 cell tumor-implanted mouse model. [Figure 8] FIG. 1 shows the effects of compound 2 and PLX3397 on the proportion of M1 macrophages in the tumor microenvironment after administration to a 4T1 cell tumor-implanted mouse model. [Figure 9] FIG. 1 shows the effects of compound 2 and PLX3397 on the proportion of M2 macrophages in the tumor microenvironment after administration to a 4T1 cell tumor-implanted mouse model. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0018] term Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art of the protected subject matter.

[0019] Unless otherwise indicated, conventional methods within the skill of those in the art, such as mass spectrometry, NMR, HPLC, protein chemistry, biochemistry, recombinant DNA technology and pharmacology, are employed in the present invention. Unless specific definitions are provided, the nomenclature associated with chemistry, such as analytical chemistry, synthetic organic chemistry, and medicinal chemistry, described herein, as well as the laboratory procedures and techniques of these chemistries, are known to those of skill in the art. In general, the techniques and procedures described above can be carried out by conventional methods well known in the art and described in the various general and more specialized references referenced and described throughout this specification.

[0020] The term "alkyl" refers to an aliphatic hydrocarbon group, which may be a branched or straight chain alkyl. Depending on the structure, the alkyl may be a monovalent group or a divalent group (i.e., an alkylene group). In the present invention, the alkyl group is preferably an alkyl having 1 to 8 carbon atoms, more preferably a "lower alkyl" having 1 to 6 carbon atoms, and even more preferably an alkyl having 1 to 4 carbon atoms. Typical alkyl groups include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, and the like. It should be understood that the "alkyl" referred to herein encompasses all possible configurations and conformations of alkyl, for example, the "propyl" referred to herein is intended to encompass n-propyl and isopropyl, the "butyl" encompasses n-butyl, isobutyl, and tert-butyl, and the "pentyl" encompasses n-pentyl, isopentyl, neopentyl, tert-pentyl, and pent-3-yl.

[0021] The term "alkoxy" refers to an -O-alkyl group, where alkyl is as defined herein. Typical alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, pentyloxy, hexyloxy, and the like.

[0022] The term "cycloalkyl" refers to a monocyclic or polycyclic radical containing only carbon and hydrogen. Cycloalkyl groups include groups having 3 to 12 ring atoms. Depending on the structure, cycloalkyl groups may be monovalent or divalent (e.g., cycloalkylene groups). In the present invention, cycloalkyl groups are preferably cycloalkyl groups having 3 to 8 carbon atoms, more preferably "lower cycloalkyl" groups having 3 to 6 carbon atoms. Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, and adamantyl.

[0023] The term "aromatic group" refers to a planar ring having a delocalized π-electron system containing 4n+2 π-electrons, where n is an integer. An aromatic ring may be formed by 5, 6, 7, 8, 9, or 10 or more atoms. An aromatic group may be optionally substituted. The term "aromatic group" includes both carbocyclic aryl (e.g., phenyl) and heterocyclic aryl (or "heteroaryl" or "heteroaromatic") groups (e.g., pyridine). The term includes monocyclic or fused-ring polycyclic (i.e., rings that share adjacent pairs of carbon atoms) groups.

[0024] As used herein, the term "aryl" refers to an aromatic ring in which each atom forming the ring is a carbon atom. The aryl ring may be formed by 5, 6, 7, 8, 9, or 10 or more atoms. The aryl group may be optionally substituted. Examples of aryl groups include, but are not limited to, phenyl, naphthalenyl, phenanthrenyl, anthracenyl, fluorenyl, and indenyl. Depending on the structure, the aryl group may be a monovalent group or a divalent group (i.e., an arylene group).

[0025] The term "aryloxy" refers to the group --O-aryl, where aryl is as defined herein.

[0026] The term "heteroaryl" refers to an aryl group that contains one or more ring heteroatoms selected from nitrogen, oxygen, and sulfur. An N-containing "heteroaryl" moiety refers to an aromatic group in which at least one of the skeletal atoms of the ring is a nitrogen atom. Depending on the structure, a heteroaryl group can be a monovalent or divalent group (i.e., a heteroarylene group). Examples of heteroaryl groups include, but are not limited to, pyridinyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, indolyl, benzimidazolyl, benzofuranyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl, isoindolyl, pteridinyl, purinyl, oxadiazolyl, thiadiazolyl, furazanyl, benzofurazanyl, benzothienyl, benzothiazolyl, benzoxazolyl, quinazolinyl, naphthyridinyl, furopyridinyl, and the like.

[0027] As used herein, the term "heteroalkyl" refers to an alkyl group, as defined herein, in which one or more of the skeletal atoms is a heteroatom, such as oxygen, nitrogen, sulfur, silicon, phosphorus, or combinations thereof. The heteroatom(s) may be placed at any interior position of the heteroalkyl group or at the position at which the heteroalkyl group is attached to the remainder of the molecule.

[0028] As used herein, the term "heterocycloalkyl" or "heterocyclyl" refers to a non-aromatic ring in which one or more of the atoms forming the ring is a heteroatom selected from the group consisting of nitrogen, oxygen, and sulfur. The heterocycloalkyl ring may be formed by 3, 4, 5, 6, 7, 8, 9, or 10 or more atoms. The heterocycloalkyl ring may be optionally substituted. Examples of heterocycloalkyls include lactams, lactones, cyclic imides, cyclic thioimides, cyclic carbamates, tetrahydrothiopyrans, 4H-pyrans, tetrahydropyrans, piperidines, 1,3-dioxines, 1,3-dioxanes, 1,4-dioxanes, 1,4-dioxanes, piperazines, 1,3-oxathianes, 1,4-oxathiines, 1,4-oxathianes, tetrahydro-1,4-thiazines, 2H-1,2-oxazines, maleimides, succinimides, barbituric acids, thiobarbituric acids, dioxopiperazines, hydantoins, dihyridines, and the like. These include, but are not limited to, dolauracil, morpholine, trioxane, hexahydro-1,3,5-triazine, tetrahydrothiophene, tetrahydrofuran, pyrroline, pyrrolidine, imidazolidine, pyrrolidone, pyrazoline, pyrazolidine, imidazoline, imidazolidine, 1,3-dioxole, 1,3-dioxolane, 1,3-dithiole, 1,3-dithiolane, isoxazoline, isoxazolidine, oxazoline, oxazolidine, oxazolidinone, thiazoline, thiazolidine, and 1,3-oxathiolane. Depending on the structure, heterocycloalkyl groups can be monovalent or divalent groups (i.e., heterocycloalkylene groups).

[0029] The term "halo" or "halogen" means fluoro, chloro, bromo and iodo.

[0030] The terms "haloalkyl," "haloalkoxy," and "haloheteroalkyl" include alkyl, alkoxy, or heteroalkyl structures in which at least one hydrogen has been replaced with a halogen atom. In certain embodiments in which two or more hydrogen atoms have been replaced with halogen atoms, the halogen atoms are the same or different from one another.

[0031] The term "hydroxy" refers to an --OH group.

[0032] The term "cyano" refers to the group -CN.

[0033] The term "ester group" refers to a chemical moiety of formula -COOR, where R is selected from alkyl, cycloalkyl, aryl, heteroaryl (bonded through a ring carbon), and heterocyclyl (bonded through a ring carbon).

[0034] The term "amino" means -NH 2 Refers to the base.

[0035] The term "aminoacyl" means -CO-NH 2 Refers to the base.

[0036] The term "amido" or "acylamino" refers to the group -NR-CO-R', where R and R' are each independently hydrogen or alkyl.

[0037] The term "optionally" means that the event(s) described below may or may not occur, and includes both the occurrence of the event(s) and the absence of the event(s). The term "optionally substituted" or "substituted" means that the referenced group may be substituted with one or more additional groups individually and independently selected from the group consisting of alkyl, cycloalkyl, aryl, heteroaryl, heterocyclyl, hydroxy, alkoxy, cyano, halo, amido, nitro, haloalkyl, amino, mesyl, alkylcarbonyl, alkoxycarbonyl, heteroarylalkyl, heterocycloalkylalkyl, aminoacyl, amino protecting groups, and the like. Of these, amino protecting groups are preferably selected from the group consisting of pivaloyl, tert-butoxycarbonyl, benzyloxycarbonyl, 9-fluorenylmethoxycarbonyl, benzyl, p-methoxybenzyl, allyloxycarbonyl, trifluoroacetyl, and the like.

[0038] As used herein, the term "tyrosine protein kinase (TPK)" refers to a type of kinase that catalyzes the transfer of the γ-phosphate from adenosine triphosphate (ATP) to tyrosine residues in proteins and can catalyze the phosphorylation of tyrosine residues in a variety of protein substrates, thus having important effects on cell growth, proliferation and differentiation.

[0039] As used herein, the terms "inhibit," "inhibition," or "inhibitor" of a kinase refer to the inhibition of phosphotransferase activity.

[0040] A "metabolite" of a compound disclosed herein is a derivative of the compound that is formed when the compound is metabolized. The term "active metabolite" refers to a biologically active derivative of a compound that is formed when the compound is metabolized. As used herein, the term "metabolized" refers to a summary of processes by which a particular substance is transformed by an organism, including, but not limited to, hydrolysis reactions and reactions catalyzed by enzymes, such as oxidation reactions. Thus, enzymes may cause a particular structural change in a compound. For example, cytochrome P450 catalyzes a variety of oxidation and reduction reactions, while uridine diphosphate glucuronyltransferase catalyzes the transfer of activated glucuronic acid molecules to aromatic alcohols, aliphatic alcohols, carboxylic acids, amines, and free sulfhydryl groups. Further information regarding metabolism can be obtained from The Pharmacological Basis of Therapeutics, 9th Edition, McGraw-Hill (1996). Metabolites of a compound disclosed herein can be identified by either administering the compound to a host and analyzing tissue samples from the host, or by incubating the compound with liver cells in vitro and analyzing the resulting compound. Both methods are well known in the art. In some embodiments, the metabolites of the compound are formed by oxidation processes and correspond to the corresponding hydroxy-containing compounds. In some embodiments, the compound is metabolized to a pharmacologically active metabolite. As used herein, the term "modulate" refers to altering the activity of a target by interacting with the target either directly or indirectly, including, by way of example only, enhancing the activity of the target, inhibiting the activity of the target, limiting the activity of the target, or prolonging the activity of the target.

[0041] As used herein, IC 50 refers to the amount, concentration or dosage of a particular test compound that achieves 50% inhibition of a maximal effect in an assay that measures such effect.

[0042] As used herein, EC 50 refers to the dose, concentration or amount of a test compound that elicits a dose-dependent response at 50% of the maximal expression of a particular response induced, stimulated or enhanced by a particular test compound.

[0043] As used herein, GI 50 refers to the concentration of a drug required to inhibit cell growth by 50%, i.e., the drug concentration at which 50% of the growth of cells (such as cancer cells) is inhibited or controlled.

[0044] The novel kinase inhibitors of the present invention The present invention relates to compounds of formula (I), or a pharma- ceutically acceptable salt, solvate, ester, acid, metabolite or prodrug thereof: [ka] (In the formula, Y is [ka] is selected from A is selected from aryl or 6-membered heterocyclyl; n is an integer selected from 1 to 3; R 1 is hydrogen, halo, C 1~6 Alkyl, C 1~6 Haloalkyl, C 1~6 selected from alkoxy and cyano; R 2 and R 3 each independently represents hydrogen, halo, C 1~6 Alkyl, C 1~6 Haloalkyl, C 1~6 alkoxy or R 2 and R 3 taken together form a phenyl or a 5-membered heterocyclyl.

[0045] In a preferred embodiment, Y is [ka] When Y is [ka] When R is , n is preferably 3. In another embodiment, A is preferably selected from phenyl, N-morpholinyl, N-piperidyl, or N-piperazinyl. More preferably, R 1 is selected from hydrogen, fluoro, chloro, methyl, ethyl, propyl, or cyano; R 2 and R 3 are each independently selected from hydrogen, fluoro, chloro, methyl, ethyl, propyl, trifluoromethyl, difluoromethyl, trifluoroethyl, methoxy, ethoxy, propoxy, or R 2 and R 3 combine to form a phenyl or dioxolane.

[0046] In a preferred embodiment, the present invention provides a compound of formula (Ia), or a pharma- ceutically acceptable salt, solvate, ester, acid, metabolite or prodrug thereof: [ka] (In the formula, R 1 , R 2 and R 3 is defined as above).

[0047] More preferably, in the formula, R 1 is selected from hydrogen, fluoro, chloro, and methyl; R 2 and R 3 are each independently selected from hydrogen, fluoro, chloro, methyl, and trifluoromethyl, or R 2 and R 3 combine to form phenyl.

[0048] In a further preferred embodiment, the present invention relates to a compound of formula (Ib), or a pharma- ceutically acceptable salt, solvate, ester, acid, metabolite or prodrug thereof: [ka] (In the formula, A, R 1 , R 2 and R 3 is defined as above).

[0049] More preferably, A is selected from phenyl or N-morpholinyl, and R 1 is hydrogen, and R 2 and R 3 is each independently selected from hydrogen, fluoro, chloro, methyl, and trifluoromethyl.

[0050] In a preferred embodiment, the present invention relates to a compound in Table 1 below, or a pharma- ceutically acceptable salt, solvate, ester, acid, metabolite or prodrug thereof.

[0051] [Table 1] TIFF2025507088000015.tif249149TIFF2025507088000016.tif103145

[0052] Any combination of the groups described above for the various variables is contemplated herein. It is understood that the substituents and substitution patterns of the compounds presented herein can be selected by one of ordinary skill in the art to provide compounds that are chemically stable and that can be synthesized by techniques known in the art, not just those described herein.

[0053] Pharmaceutically acceptable salts, solvates, esters, acids, pharma- ceutically active metabolites and prodrugs of these compounds are also described herein.

[0054] In additional or further embodiments, the compounds described herein, when administered to an organism in need thereof, are metabolized to produce metabolites which are then used to provide a desired effect, including a desired therapeutic effect.

[0055] The compounds described herein may be formed and / or used as pharma- ceutically acceptable salts. Types of pharma-ceutically acceptable salts include: (1) salts of the free base form of the compound with pharma- ceutically acceptable inorganic acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, metaphosphoric acid, or salts of acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, malic acid, citric acid, succinic acid, maleic acid, tartaric acid, fumaric acid, trifluoroacetic acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, Mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, toluenesulfonic acid, 4-methylbicyclo-[2.2.2]oct-2-ene-1-carboxylic acid, 2-naphthalenesulfonic acid, tert-butylacetic acid, glucoheptonic acid, 4,4'-methylenebis-(3-hydroxy-2-ene-1-carboxylic acid), 3-phenylpropionic acid, trimethylacetic acid, la (2) base addition salts formed when the acidic hydrogen ion present in the parent compound is replaced by a metal ion, such as an alkali metal ion (e.g., lithium, sodium, potassium), alkaline earth metal ion (e.g., magnesium or calcium), or aluminum ion, or when coordinated with an organic or inorganic base. Acceptable organic bases include, but are not limited to, ethanolamine, diethanolamine, triethanolamine, trimethylamine, N-methylglucamine, and the like, and acceptable inorganic bases include aluminum hydroxide, calcium hydroxide, potassium hydroxide, sodium carbonate, sodium hydroxide, and the like.

[0056] The corresponding counter ions of pharma- ceutically acceptable salts can be analyzed and identified using a variety of methods, including, but not limited to, ion exchange chromatography, ion chromatography, capillary electrophoresis, inductively coupled plasma, atomic absorption spectroscopy, mass spectrometry, or any combination thereof.

[0057] The salts are recovered using at least one of the following techniques: filtration, precipitation with a non-solvent followed by filtration, evaporation of the solvent, or, in the case of aqueous solutions, lyophilization.

[0058] Screening and characterization of pharma-ceutically acceptable salts, polymorphs and / or solvates can be accomplished using a variety of techniques, including, but not limited to, thermal analysis, x-ray diffraction, spectroscopy, microscopy, and elemental analysis. The various spectroscopic techniques used include, but are not limited to, Raman, FTIR, UVIS, and NMR (liquid and solid). The various microscopic techniques include, but are not limited to, IR microscopy and Raman microscopy.

[0059] Medical uses of the present invention The compounds of formula (I), formula (Ia) and formula (Ib) of the present invention, or pharma- ceutically acceptable salts, solvates, esters, acids, metabolites or prodrugs thereof, can convert macrophages in the TME from M2, which promotes tumor growth, to M1, thereby changing the TME and achieving the purpose of treating cancer. Furthermore, the compounds of formula (I), formula (Ia) and formula (Ib) of the present invention, or pharma-ceutically acceptable salts, solvates, esters, acids, metabolites or prodrugs thereof, can inhibit the activity of CSF1R kinase and related signal transduction pathways, thereby achieving the purpose of treating cancer.

[0060] Thus, the compounds of formula (I), formula (Ia) and formula (Ib) of the present invention, or pharma- ceutically acceptable salts, solvates, esters, acids, metabolites or prodrugs thereof, can be used to treat cancers or tumors in which M2 macrophages predominate in the tumor microenvironment.

[0061] The cancer or tumor is selected from one or more of ovarian cancer, cervical cancer, membranous adenocarcinoma, prostate cancer, bladder cancer, lung cancer, thyroid cancer, breast cancer, pancreatic cancer, kidney cancer, stomach cancer, liver cancer, cervical cancer, endometrial cancer, colorectal cancer, nasopharyngeal cancer, esophageal cancer, bile duct cancer, bone metastasis cancer, papillary thyroid cancer, non-small cell lung cancer, small cell lung cancer, colon cancer, solid tumor, brain tumor, lymphoma, glioma, glioblastoma, melanoma, mesothelioma, glioblastoma, osteosarcoma, gastrointestinal stromal tumor, multiple myeloma, bile duct carcinoma sarcoma, and leukemia. Among these, lymphoma includes non-Hodgkin's lymphoma, B-cell or T-cell lymphoma, and leukemia includes chronic myelogenous leukemia or acute myelogenous leukemia.

[0062] In an embodiment of the present invention, a pharmaceutical agent comprising a compound of the present invention can be administered to a patient by at least one of injection, oral administration, inhalation, rectal administration, and transdermal administration. When treating a patient according to the present invention, the amount of a given agent will depend on many factors, such as the particular administration regimen, the type of disease or condition and its severity, the identity of the subject or host requiring treatment (e.g., body weight), and the dose administered can be routinely determined by methods known in the art depending on the particular circumstances surrounding the case, including, for example, the particular agent administered, the route of administration, the condition being treated, and the subject or host being treated. In general, doses used for adult treatment will usually take the range of 0.02 mg to 5000 mg per day, for example, about 1 mg to 1500 mg per day. The desired dose may conveniently be presented as a single dose or as divided doses administered simultaneously (or closely spaced) or at suitable intervals, for example, as two, three, four or more divided doses per day. Those skilled in the art will recognize that even though the above dosage ranges are given, the specific effective amount may be appropriately adjusted depending on the patient's condition and the physician's judgment.

[0063] Preparation of compounds Compounds of formula (I) can be synthesized using standard synthetic techniques known to those skilled in the art, or using methods known in the art in combination with the methods described herein. Furthermore, the solvents, temperatures and other reaction conditions presented herein may vary according to those skilled in the art. As a further guide, the following synthetic methods may also be used.

[0064] The reactions can be used sequentially to provide the compounds described herein, or can be used to synthesize fragments that are subsequently coupled by methods described herein and / or known in the art.

[0065] In certain embodiments, methods for preparing the tyrosine kinase inhibitor compounds described herein and methods for using them are provided herein. In certain embodiments, the compounds described herein can be synthesized using the following synthetic schemes. The compounds can be synthesized using methods similar to those described below by using appropriate alternative starting materials.

[0066] The starting materials used in the synthesis of the compounds described herein may be synthesized or obtained from commercial sources. The compounds described herein, and other related compounds with different substituents, can be synthesized using techniques and materials known to those skilled in the art. The general methods for preparing the compounds disclosed herein can be derived from reactions known in the art, and the reactions can be modified by using appropriate reagents and conditions to introduce various moieties into the molecules presented herein, as recognized by those skilled in the art.

[0067] The reaction products can be isolated and purified, if desired, using conventional techniques, including but not limited to filtration, distillation, crystallization, chromatography, etc. Such products can be characterized using conventional means, including physical constants and spectral data.

[0068] Non-limiting examples of synthetic schemes for preparing compounds of formula (I) are shown in the synthetic routes below. EXAMPLES

[0069] Example 1: N-(4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-2-(4-methyl-3-(trifluoromethyl)phenyl)acetamide [ka] 4-((6,7-Dimethoxyquinolin-4-yl)oxy)phenylamine (A2): 4-Hydroxyphenylamine (2.0 g) was added to a round bottom flask, followed by dimethylsulfoxide (30 ml), and then sodium hydride (807 mg) was added dropwise under ice bath conditions. The reaction was stirred at room temperature for 30 minutes. Then, 4-chloro-6,7-dimethoxyquinoline (4.1 g) was added and the reaction was allowed to react at 90° C. overnight. After completion of the reaction, the system was added with water, filtered, and the solid was washed with water and a small amount of methanol. The brown solid obtained was used directly in the next step.

[0070] N-(4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-2-(4-methyl-3-(trifluoromethyl)phenyl)acetamide (1): 4-((6,7-dimethoxyquinolin-4-yl)oxy)phenylamine (1 g) was added to a round bottom flask, followed by N,N-dimethylformamide (10 ml), HATU (1.92 g), 4-methyl-3-trifluoromethylphenylacetic acid (1.10 g) and N,N-diisopropylethylamine (0.87 g). The reaction was stirred at room temperature overnight. After completion of the reaction, water was added to the system, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate. The organic phase was filtered, the solvent was removed from the filtrate under reduced pressure, and the residue was purified by pressurized silica gel column chromatography to give compound 1. LC / MS: M+H 497.1.

[0071] Example 2: N-(4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-2-(4-chloro-3-(trifluoromethyl)phenyl)-acetamide [ka] The synthesis of the compound of Example 2 was completed by using a procedure similar to that of Example 1. MS (ESI) m / z (M+1)+: 517.11.

[0072] Example 3: N-(4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-2-phenylacetamide [ka] The synthesis of the compound of Example 3 was completed by using a procedure similar to that of Example 1. MS (ESI) m / z (M+1)+: 415.16.

[0073] Example 4: N-(4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-3-(piperidin-1-yl)propanamide [ka] The synthesis of the compound of Example 4 was completed by using a procedure similar to that of Example 1. MS (ESI) m / z (M+1)+: 436.22.

[0074] Example 5: N-(4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-2-(4-chloro-3-fluorophenyl)acetamide [ka] The synthesis of the compound of Example 5 was completed by using a procedure similar to that of Example 1. MS (ESI) m / z (M+1)+: 467.11.

[0075] Example 6: N-(2-chloro-4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-2-(3-(trifluoromethyl)phenyl)acetamide [ka] The synthesis of the compound of Example 6 was completed by using a procedure similar to that of Example 1. MS (ESI) m / z (M+1)+: 517.11.

[0076] Example 7: N-(2-chloro-4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-2-(4-(trifluoromethyl)phenyl)acetamide [ka] The synthesis of the compound of Example 7 was completed by using a procedure similar to that of Example 1. MS (ESI) m / z (M+1)+: 517.11.

[0077] Example 8: N-(4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-2-(3-(trifluoromethyl)phenyl)acetamide [ka] The synthesis of the compound of Example 8 was completed by using a procedure similar to that of Example 1. MS (ESI) m / z (M+1)+: 483.15.

[0078] Example 9: N-(4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-2-(4-(trifluoromethyl)phenyl)acetamide [ka] The synthesis of the compound of Example 9 was completed by using a procedure similar to that of Example 1. MS (ESI) m / z (M+1)+: 483.15.

[0079] Example 10: N-(4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-2-(3,4-dimethoxyphenyl)acetamide [ka] The synthesis of the compound of Example 10 was completed by using a procedure similar to that of Example 1. MS (ESI) m / z (M+1)+: 475.18.

[0080] Example 11: N-(4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)2-(3,4-dimethylphenyl)acetamide [ka] The synthesis of the compound of Example 11 was completed by using a procedure similar to that of Example 1. MS (ESI) m / z (M+1)+: 443.19.

[0081] Example 12: N-(4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-2-(3,4-difluorophenyl)acetamide [ka] The synthesis of the compound of Example 12 was completed by using a procedure similar to that of Example 1. MS (ESI) m / z (M+1)+: 451.14.

[0082] Example 13: N-(4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-2-(4-fluoro-3-(trifluoromethyl)phenyl)acetamide [ka] The synthesis of the compound of Example 13 was completed by using a procedure similar to that of Example 1. MS (ESI) m / z (M+1)+: 501.14.

[0083] Example 14: N-(4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-2-(2,6-difluorophenyl)acetamide [ka] The synthesis of the compound of Example 14 was completed by using a procedure similar to that of Example 1. MS (ESI) m / z (M+1)+: 451.14.

[0084] Example 15: N-(4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-2-(2,4-difluorophenyl)acetamide [ka] The synthesis of the compound of Example 15 was completed by using a procedure similar to that of Example 1. MS (ESI) m / z (M+1)+: 451.14.

[0085] Example 16: N-(4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-2-(3-fluoro-4-methylphenyl)acetamide [ka] The synthesis of the compound of Example 16 was completed by using a procedure similar to that of Example 1. MS (ESI) m / z (M+1)+: 447.17.

[0086] Example 17: N-(4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-2-(3,4-dichlorophenyl)acetamide [ka] The synthesis of the compound of Example 17 was completed by using a procedure similar to that of Example 1. MS (ESI) m / z (M+1)+: 483.08.

[0087] Example 18: N-(4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-2-(benzo[d][1,3]dioxol-5-yl)acetamide [ka] The synthesis of the compound of Example 18 was completed by using a procedure similar to that of Example 1. MS (ESI) m / z (M+1)+: 459.15.

[0088] Example 19: N-(4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-2-(naphth-2-yl)acetamide [ka] The synthesis of the compound of Example 19 was completed by using a procedure similar to that of Example 1. MS (ESI) m / z (M+1)+: 465.18.

[0089] Example 20: N-(4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-2-(3-fluorophenyl)acetamide [ka] The synthesis of the compound of Example 20 was completed by using a procedure similar to that of Example 1. MS (ESI) m / z (M+1)+: 433.15.

[0090] Example 21: N-(4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-2-(4-methylpiperazin-1-yl)acetamide [ka] The synthesis of the compound of Example 21 was completed by using a procedure similar to that of Example 1. MS (ESI) m / z (M+1)+: 437.21.

[0091] Example 22: N-(4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-3-(4-methylpiperazin-1-yl)propanamide [ka] The synthesis of the compound of Example 22 was completed by using a procedure similar to that of Example 1. MS (ESI) m / z (M+1)+: 451.23.

[0092] Example 23: N-(2-chloro-4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-2-(4-chloro-3-(trifluoromethyl)phenyl)acetamide [ka] The synthesis of the compound of Example 23 was completed by using a procedure similar to that of Example 1. MS (ESI) m / z (M+1)+: 551.07.

[0093] Example 24: N-(2-fluoro-4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-2-(4-chloro-3-(trifluoromethyl)phenyl)acetamide [ka] The synthesis of the compound of Example 24 was completed by using a procedure similar to that of Example 1. MS (ESI) m / z (M+1)+: 535.10.

[0094] Example 25: N-(3-fluoro-4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-2-(4-chloro-3-(trifluoromethyl)phenyl)acetamide [ka] The synthesis of the compound of Example 25 was completed by using a procedure similar to that of Example 1. MS (ESI) m / z (M+1)+: 535.10.

[0095] Example 26: N-(2-methyl-4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-2-(4-chloro-3-(trifluoromethyl)phenyl)acetamide [ka] The synthesis of the compound of Example 26 was completed by using a procedure similar to that of Example 1. MS (ESI) m / z (M+1)+: 531.12.

[0096] Example 27: N-(3-methyl-4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-2-(4-chloro-3-(trifluoromethyl)phenyl)acetamide [ka] The synthesis of the compound of Example 27 was completed by using a procedure similar to that of Example 1. MS (ESI) m / z (M+1)+: 531.12.

[0097] Example 28: 1-(4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-3-(3-morpholinopropyl)urea [ka] Phenyl(4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)carbamate (G1): 4-((6,7-dimethoxyquinolin-4-yl)oxy)phenylamine (1 g) was dissolved in dichloromethane (30 ml), followed by the addition of pyridine (0.4 g) and dropwise addition of phenyl chloroformate (0.8 g) under ice bath conditions. After the addition, stirring was continued for 3 hours. After the reaction was completed, water was added to the reaction system and extracted with dichloromethane. The organic phase was dried over anhydrous sodium sulfate and filtered. Dichloromethane was removed under reduced pressure and the residue was purified by pressurized silica gel column chromatography to obtain compound G1. MS: (M+1) 417.14.

[0098] 1-(4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-3-(3-morpholinopropyl)urea (42): In a round bottom flask, phenyl(4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)carbamate (50 mg), N-(3-aminopropyl)morpholine (26 mg), N,N-diisopropylethylamine (23 mg), and tetrahydrofuran (3 ml) were added. The reaction mixture was reacted at 80° C. overnight. After completion of the reaction, the reaction was concentrated and the residue was purified by pressurized silica gel column chromatography to give compound 28. MS: (M+1) 467.22.

[0099] Example 29: 1-(4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-3-(3-(4-methylpiperazin-1-yl)propyl)urea [ka] The synthesis of the compound of Example 29 was completed by using a procedure similar to that of Example 28. MS (ESI) m / z (M+1)+: 480.26.

[0100] Example 30: 1-(4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-3-(3-phenylpropyl)urea [ka] The synthesis of example 30 compound was completed by using a procedure similar to that of example 28. MS (ESI) m / z (M+1)+: 458.20.

[0101] Example 31: 1-(4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-3-(3-(3-fluorophenyl)propyl)urea [ka] The synthesis of the compound of Example 31 was completed by using a procedure similar to that of Example 28. MS (ESI) m / z (M+1)+: 476.19.

[0102] Example 32: 1-(4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-3-(3-(4-fluorophenyl)propyl)urea [ka] The synthesis of the compound of Example 32 was completed by using a procedure similar to that of Example 28. MS (ESI) m / z (M+1)+: 476.19.

[0103] Example 33: 1-(4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-3-(3-(3-(trifluoromethyl)phenyl)propyl)urea [ka] The synthesis of the compound of Example 33 was completed by using a procedure similar to that of Example 28. MS (ESI) m / z (M+1)+: 526.19.

[0104] Example 34: (1-(4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-3-(3-(3-(methyl)phenyl)propyl)urea [ka] The synthesis of example 34 was completed by using a procedure similar to that of example 28. MS (ESI) m / z (M+1)+: 472.22.

[0105] Example 35: 1-(4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-3-(3-(4-(trifluoromethyl)phenyl)propyl)urea [ka] The synthesis of the compound of Example 35 was completed by using a procedure similar to that of Example 28. MS (ESI) m / z (M+1)+: 526.19.

[0106] Example 36: N-(3-trifluoromethyl-4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-2-(4-chloro-3-(trifluoromethyl)phenyl)acetamide [ka] The synthesis of the compound of Example 36 was completed by using a procedure similar to that of Example 1. MS (ESI) m / z (M+1)+: 585.10.

[0107] Example 37: N-(3-methoxy-4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-2-(4-chloro-3-(trifluoromethyl)phenyl)acetamide [ka] The synthesis of the compound of Example 37 was completed by using a procedure similar to that of Example 1. MS (ESI) m / z (M+1)+: 547.12.

[0108] Example 38: N-(3-chloro-4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-2-(4-chloro-3-(trifluoromethyl)phenyl)acetamide [ka] The synthesis of the compound of Example 38 was completed by using a procedure similar to that of Example 1. MS (ESI) m / z (M+1)+: 551.08.

[0109] Example 39: N-(3-cyano-4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-2-(4-chloro-3-(trifluoromethyl)phenyl)acetamide [ka] The synthesis of the compound of Example 39 was completed by using a procedure similar to that of Example 1. MS (ESI) m / z (M+1)+: 542.11.

[0110] Example 40: N-(4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-2-(2-(trifluoromethyl)phenyl)acetamide [ka] The synthesis of example 40 compound was completed by using a procedure similar to that of example 1. MS (ESI) m / z (M+1)+: 483.15.

[0111] Comparative Example 1: N-(6-((6,7-dimethoxyquinolin-4-yl)oxy)pyridin-3-yl)-2-phenylacetamide [ka] 6,7-Dimethoxy-4-((5-nitropyridin-2-yl)oxy)quinoline (D2): 4-Hydroxy-6,7-dimethoxyquinoline (0.5 g) was added to a round bottom flask, followed by acetonitrile (10 ml), and then cesium carbonate (0.87 g). The mixture was stirred at room temperature for 5 minutes. Then, 2-chloro-5-nitropyridine (0.42 g) was added and stirred at room temperature overnight. After completion of the reaction, it was filtered, the filtrate was concentrated, and the residue was purified by pressurized silica gel column chromatography to obtain compound D2.

[0112] 6-((6,7-Dimethoxyquinolin-4-yl)oxy)pyridin-3-amine (D3): 6,7-Dimethoxy-4-((5-nitropyridin-2-yl)oxy)quinoline (0.42 g) was added to a round bottom flask, followed by methanol (10 ml) and palladium on carbon (0.11 g). The reaction was carried out overnight under hydrogen atmosphere. After completion of the reaction, it was filtered, the filtrate was concentrated and the residue was purified by pressurized silica gel column chromatography to give compound D3. LC / MS: M+H 298.11.

[0113] N-(6-((6,7-dimethoxyquinolin-4-yl)oxy)pyridin-3-yl)-2-phenylacetamide (Comparative Compound 1): 6-((6,7-dimethoxyquinolin-4-yl)oxy)pyridin-3-amine (50 mg) was added to a round bottom flask, followed by N,N-dimethylformamide (2 ml), HATU (96 mg), phenylacetic acid (34 mg), and N,N-diisopropylethylamine (66 mg). The reaction was stirred at room temperature overnight. After completion of the reaction, water was added to the system, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate. The organic phase was filtered, the solvent was removed from the filtrate under reduced pressure, and the residue was purified by pressurized silica gel column chromatography to obtain Comparative Compound 1. LC / MS: M+H 416.16.

[0114] Comparative Example 2: N-(5-chloro-6-((6,7-dimethoxyquinolin-4-yl)oxy)pyridin-3-yl)-2-(3-(trifluoromethyl)phenyl)acetamide [ka] The synthesis of the compound of Comparative Example 2 was completed by using a similar procedure as that of Comparative Example 1. MS (ESI) m / z (M+1)+: 518.10.

[0115] Comparative Example 3: N-(3-(trifluoromethyl)benzyl)-4-((6,7-dimethoxyquinolin-4-yl)oxy)benzamide [ka] Methyl 4-((6,7-dimethoxyquinolin-4-yl)oxy)benzoate (E2): In a round bottom flask, 4-chloro-6,7-dimethoxyquinoline (1.0 g) and methyl 4-hydroxybenzoate (0.68 g) were added, and the mixture was stirred at 145° C. overnight. After completion of the reaction, the mixture was added with saturated sodium bicarbonate solution, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated, and the residue was purified by pressurized silica gel column chromatography to obtain compound E2.

[0116] 4-((6,7-dimethoxyquinolin-4-yl)oxy)benzoic acid (E3): To a round bottom flask was added methyl 4-((6,7-dimethoxyquinolin-4-yl)oxy)benzoate (0.5 g), methanol / tetrahydrofuran (5:1, 10 ml), and lithium hydroxide (0.18 g). The reaction was carried out at room temperature overnight. After completion of the reaction, it was concentrated and the pH was adjusted to about 5 with concentrated hydrochloric acid. It was filtered and the filter cake was washed with water to obtain compound E3. LC / MS: M+H 326.10.

[0117] N-(3-(trifluoromethyl)benzyl)-4-((6,7-dimethoxyquinolin-4-yl)oxy)benzamide (Comparative Compound 3): 4-((6,7-dimethoxyquinolin-4-yl)oxy)benzoic acid (20 mg) was added to a round bottom flask, followed by N,N-dimethylformamide (2 ml), 3-trifluoromethylbenzylamine (13 mg), EDCI (17 mg), HOBT (12 mg), and triethylamine (12 mg). The reaction was stirred at room temperature overnight. After completion of the reaction, water was added to the system, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate. The organic phase was filtered, and the solvent was removed from the filtrate under reduced pressure. The residue was purified by pressurized silica gel column chromatography to obtain Comparative Compound 3. LC / MS: M+H 483.15.

[0118] Comparative Example 4: N-(4-((6,7-dimethoxyquinazolin-4-yl)oxy)phenyl)-2-(3-methoxyphenyl)acetamide [ka] The synthesis of the compound of Comparative Example 4 was completed by using a procedure similar to that of Example 1. MS (ESI) m / z (M+1)+: 446.17.

[0119] Example 41: Effect on cancer cell proliferation The inhibitory effect of the compounds presented herein on the proliferation of cancer cells was further evaluated by testing the effect of the compounds of the present invention on the growth of cancer cells.

[0120] In this example, various concentrations (0.000508μM, 0.00152μM, 0.00457μM, 0.0137μM, 0.0411μM, 0.123μM, 0.370μM, 1.11μM, 3.33μM, 10μM in DMSO) of the compounds of the present invention and comparative compounds 1-4 were added to TEL-CSF1R-BaF3 (constructed in our laboratory) and M-NSF-60 (purchased from COBIOER BIOSCIENCES CO., LTD (Nanjing, China), M-CSF (CSF-1) mediated mouse myeloblast cell line, when this cell line is used to evaluate the inhibitory effect of the compounds on the proliferation of cancer cells, 50ng / mL of M-CSF was added to the cell culture medium) and incubated for 72 hours. The incubated cells were detected by CCK-8 (purchased from MedChem Express, China) cell viability detection kit (CCK-8 can be reduced to a highly water-soluble yellow formazan product by dehydrogenase in viable cells, and the amount of the formazan product generated is proportional to the number of viable cells). The number of viable cells was quantified by a microplate reader, and the GI of each compound was measured. 50 was calculated (the results are shown in Table 2).

[0121] The TEL-CSF1R-BaF3 cell line (a cell line stably expressing CSF1R kinase) was constructed as follows: the sequence of the human CSF1R kinase region was amplified by PCR, inserted into the MSCV-Puro vector (Clontech) together with the N-terminal TEL fragment, and stably introduced into mouse BaF3 cells (purchased from ATCC, USA) by the retroviral method. IL-3 growth factor was then removed, and finally, a cell line dependent on the CSF1R-introduced protein was obtained.

[0122] The results in Table 2 show that the compounds of the present invention have a strong inhibitory effect on CSF1R kinase activity. Comparative Example Compounds 1 to 3 did not show an inhibitory effect on the proliferation of TEL-CSF1R-BaF3 expressing CSF1R kinase and M-NSF-60, an M-CSF-mediated mouse myeloblast cell line. Comparative Example Compound 4 showed a certain inhibitory activity on the proliferation of TEL-CSF1R-BaF3 expressing CSF1R kinase and M-NSF-60, an M-CSF-mediated mouse myeloblast cell line, but the activity was lower than that of the compounds of the present invention. The compounds of the present invention showed a strong inhibitory effect on the proliferation of TEL-CSF1R-BaF3 expressing CSF1R kinase and M-NSF-60, an M-CSF-mediated mouse myeloblast cell line. Because CSF1R kinase is a key protein that controls the polarization of macrophages in the tumor microenvironment, inhibiting the activity of CSF1R kinase can reduce the conversion of macrophages to M2 macrophages, which promote cancer development, and increase the conversion of macrophages to M1 macrophages, which inhibit cancer development.

[0123] [Table 2]

[0124] Example 42: Cell transformation experiments RAW264.7 cells (purchased from ATCC, USA) were pipetted up and down, placed in a centrifuge tube, and centrifuged at 800 rpm for 4 minutes. The cell supernatant was discarded, and 0.5 × 10 cells were collected. 5 The cells were plated at a density of 100 cells / ml in 6-well transparent plates and cultured in 1640 culture medium containing 10% FBS (1640 culture medium was purchased from Corning, USA; FBS was purchased from Excel, Australia) at 37°C and 5% CO 2The cells were cultured in an incubator for 12 hours. Then, the 6-well transparent plate was taken out, and 20ng / ml of interleukin-4 or interleukin-13 factors (both purchased from Sino Biological (Beijing, China)) were added to 1640 culture medium (purchased from Corning (USA)), and then these were added together to the cell wells of the 6-well transparent plate, and the cells were cultured in the culture medium for 24 hours. The cell morphology was checked, and when the cells gradually changed from round to spindle shape, the secretion of interleukin-10 in the supernatant of the culture medium was detected according to the instructions using a mouse IL-10 ELISA kit (purchased from Wuhan Saipei Biotechnology (China)), and the increase in the concentration of secretion of interleukin-10 suggested that the RAW264.7 cells were induced into M2 macrophages. After successful cell induction, they were treated with serially diluted randomly selected compound 2 of the present invention, control compounds BLZ945 and PLX3397 (purchased from MedChem Express (China)) for 72 h, after which the cells and supernatants were collected for qPCR and ELISA experiments, respectively.

[0125] qPCR extraction of cellular RNA In this experiment, an RNA extraction kit (purchased from Tiangen Biochemical Technology (China), Catalog No.: DP451) was used. RNA extraction was performed according to the instructions, and the concentration of RNA in the samples was detected using a NanoDrop nucleic acid concentration meter, and the RNA solution was diluted to the same concentration.

[0126] Real-time fluorescent quantitative PCR experiments In this experiment, we used the TaKaRa One Step TB Green™ PrimeScript™ PLUS RT-PCR Kit (purchased from TaKaRa Bio, catalog number: RR096A), which contains the following components:

[0127] [Table 3]

[0128] The procedure was as follows: 1. Take out the PCR primers and place them on ice. The primer sequences used were as follows:

[0129] [Table 4]

[0130] 2. The RT-PCR reaction solution was prepared on ice as follows.

[0131] [Table 5]

[0132] 3. After preparing the RT-PCR reaction reagents, they were mixed and the mixture was dispensed into 8-strip tubes. 2 μL of RNA sample was added to the tube, the bottom of the tube was tapped to mix the solution, and the tube was centrifuged in a centrifuge for a while.

[0133] 4. The fluorescent quantitative PCR device was turned on. After the device's self-check was completed, the sample was placed in the device and PCR reaction was performed according to the following procedure. The first step was reverse transcription reaction: 42°C for 5 minutes, 95°C for 10 seconds, 1 cycle. The second step was PCR amplification: 95°C for 5 seconds, 60°C for 20 seconds, 40 cycles.

[0134] 5. When the experiment was completed, the samples were removed and the device was turned off. Data was analyzed using Roche's analysis software LightCycler (trademark) 96 SW 1.1, and the analyzed data was imported into Graphpad Prism 7.0 and plotted. The results are shown in Figures 1 and 2.

[0135] The experiment showed that after the cells were induced to become M2 macrophages, the level of IL-10 secreted by M2 macrophages was much higher than that secreted by M1 macrophages, indicating that IL-10 was mainly secreted by M2 cells and less by M1 macrophages. As shown in Figure 1, after the M2 macrophages were treated with compound 2 of the present invention, the secretion level of IL-10 in M2 macrophages decreased in a dose-dependent manner with increasing drug concentration, indicating that M2 cells were gradually decreased among macrophages, and the effect of 1 μM compound 2 was better than that of the positive control PLX3397 and BLZ945 at the same dose. In addition, CD86 was a marker of M1 macrophages. As shown in Figure 2, after the cells were treated with compound 2, the RNA level of CD86 was found to increase in a dose-dependent manner with increasing drug concentration, indicating that the proportion of M1 cells in macrophages was gradually increased. The effect of 1 μM compound 2 was better than that of the positive control PLX3397 and BLZ945 at the same dose. CSF1R kinase is a key protein that controls the polarization of macrophages in tumor microenvironment, so inhibiting the activity of CSF1R kinase can reduce the conversion of macrophages to M2 macrophages that promote cancer development and increase the polarization of M1 macrophages that inhibit cancer development. This example demonstrates that the compounds involved in the present invention can reduce the conversion of macrophages to M2 macrophages and promote the conversion of macrophages to M1 macrophages, thereby inhibiting tumor growth and enhancing cancer immunotherapy.

[0136] Example 43: In vivo efficacy assay In this example, the experimental effects of Compound 2 and the control compound PLX3397 (purchased from MedChem Express (China)) in MC38 and 4T1 mouse models were tested, respectively.

[0137] The experimental procedure was as follows: (1) C57BL / 6J male mice used as MC38 transplanted tumor models and Balb / c male mice (6 weeks old) used as 4T1 transplanted tumor models were purchased from GemPharmatech Co., Ltd (Jiangsu). All the above mice were kept in an SPF laboratory, and drinking water and pads were sterilized by autoclaving. All operations related to mice were performed under aseptic conditions.

[0138] (2) On day 0, approximately 1 × 10 6 MC38 cells (purchased from ATCC) were injected subcutaneously into the left dorsal region of each C57BL / 6J mouse, giving approximately 1 × 10 6 4T1 cells (purchased from ATCC) were subcutaneously injected into the left dorsal region of each Balb / c mouse.

[0139] (3) For the MC38 mouse model, starting from day 6, the corresponding mice were orally administered methylcellulose (HKI) (purchased from China National Pharmaceutical Group Corporation) solvent once a day (5 mice), compound 2 at a dose of 50 mg per kg of mouse body weight once a day (5 mice), and PLX3397 at a dose of 50 mg per kg of mouse body weight once a day (5 mice). For the 4T1 mouse model, starting from day 6, the corresponding mice were orally administered 10% HS-15 (purchased from BASF (Germany)) solvent (5 mice), compound 2 at a dose of 50 mg per kg of mouse body weight once a day (5 mice), and PLX3397 at a dose of 50 mg per kg of mouse body weight once a day (5 mice).

[0140] (4) Starting from day 6, the length / width of the subcutaneous tumor was measured daily with a caliper and the weight of the mouse was recorded daily to investigate the effects of compound 2 and the control compound PLX3397 on the body weight of the mouse in different model mice. The results are shown in Figures 3 and 6.

[0141] (5) The growth trends of subcutaneous tumors in different mouse models were statistically analyzed. Tumor volumes were calculated using the following method: length × width × width / 2 mm3 The results are shown in Figures 4 and 7.

[0142] (6) Isolation of tumor-infiltrating lymphocytes: First, the tumor tissue was finely chopped (1 mm 3 The cells were then sieved through a 200 mesh sieve, collected in a 50 ml centrifuge tube, and centrifuged at 1500 rpm for 10 min. The supernatant was then discarded, and 10 ml of DMEM medium (10% FBS) (purchased from GIBCO, USA) was added to the centrifuge tube to stop the digestion. The centrifuge tube was placed in a centrifuge and centrifuged at 1500 rpm for another 10 min. The supernatant was discarded. At the same time, Percoll separation solution (purchased from Millipore, USA) was prepared, and 90% Percoll separation working solution was prepared using 10x PBS. To a 15 ml centrifuge tube, 3 ml to 4 ml of 70% Percoll separation solution was added. The cells were resuspended in 4 ml of 40% Percoll separation solution, and the cell mixture was added to the 70% Percoll separation solution. The centrifuge tube was placed in a centrifuge and subjected to density gradient centrifugation at 25 °C and 1260 g for 30 minutes (speed up 5, speed down 0). After centrifugation, the centrifuge tube was placed in a centrifuge tube rack, the middle white blood cell layer was aspirated with a pipette and added to 10 ml of PBS, and they were mixed well before being placed in a centrifuge and centrifuged at 1500 rpm. After 10 minutes, the cell supernatant was discarded, the cells were resuspended in 5 ml of PBS, centrifuged at 1500 rpm for another 10 minutes, and the washing process was repeated once. The obtained cells were white blood cells in the tumor tissue. The cells were placed on ice for later use.

[0143] (7) Antibody labeling for flow cytometry: Cells were labeled with three types of antibodies, first to label cell surface markers and then to label intracellular factors. Cell surface staining: 1 × 10 6The cells were harvested and centrifuged at 3500 rpm for 5 min, and the cell supernatant was discarded. 1× ​​PBS (50 μL) (purchased from Sangon Biotech Co., Ltd (China)) was added to resuspend the cells. 1 μL of CD86 antibody and 1 μL of F4 / 80 antibody (both CD86 and F4 / 80 antibodies were purchased from BD Pharmingen (USA)) were added to the centrifuge tube, and the tube was incubated at 4°C for 30 min, after which the tube was centrifuged at 3500 rpm for 5 min, and the supernatant was discarded. The cells were washed twice with 1× staining buffer, and the cells were resuspended in 100 μL of PBS for later use. Intracellular factor staining: The cells were treated with permeabilization / fixation solution. First, 250 μl of permeabilization / fixation solution (catalog number 554714, purchased from BD Biosciences (USA)) was added to the cell suspension, and the mixture was incubated on ice for 20 min. Next, 1 ml of 1x BD Perm / Wash buffer was added to resuspend the cells, and the resultant was centrifuged in a centrifuge at 3500 rpm for 5 minutes. The cell supernatant was discarded and the procedure was repeated. Then, 50 μl of 1x BD Perm / Wash buffer was added to the centrifuge tube to resuspend the cells, and an appropriate amount of 1 μl CD206 antibody (both purchased from BD Pharmingen (USA)) was added to the cell suspension, and the mixture was incubated at 4°C for 30 minutes. The procedure was repeated, and finally the cells were resuspended in PBS and detected by the machine. The data was processed using Flowjo and imported into Graphpad Prism software for plotting. The results are shown in Figure 5, Figure 8 and Figure 9.

[0144] The transplanted tumor models of C57BL / 6J mice with MC38 cells and Balb / c mice with 4T1 cells are commonly used models of tumor immunity. After administration, the drug activates the mouse's own tumor immunity, and the tumor growth is inhibited by the mouse's own immunity. The experimental results are shown in Figures 3 and 4, as well as Figures 6 and 7. Compound 2 and the control compound PLX3397 showed good inhibition effect on mouse tumors without obvious toxicity in the tumor immunity model, MC38 and 4T1 mouse tumor model. With the increase in the number of days of administration, the inhibition effect of compound 2 on mouse tumors became more and more significant, and was superior to the effect of the control compound PLX3397.

[0145] Furthermore, as shown in Figures 5, 8 and 9, by detecting the proportion of macrophages in the tumor microenvironment, it was found that the proportion of M1 macrophages around the tumor in the group administered with compound 2 was much higher than that in the vehicle group, and the proportion of M1 macrophages in the tumor microenvironment in the group administered with compound 2 at the same dose was also higher than that in the PLX3397 group. This indicates that the compounds of the present invention can induce the conversion of M2 TAM to M1 TAM to inhibit tumor growth, and therefore can be used to treat cancers or tumors in which M2 macrophages are predominant in the tumor microenvironment. [Industrial Applicability]

[0146] The present invention provides a novel use of quinoline inhibitor compounds that can polarize macrophages from M2 to M1 by inhibiting CSF1R kinase activity, thereby achieving the therapeutic effect of cancer or tumor. Therefore, the present invention is suitable for industrial application.

[0147] Although the present invention is described in detail in this specification, the present invention is not limited thereto. Those skilled in the art can make modifications according to the principle of the present invention. Therefore, any modifications made according to the principle of the present invention should be understood to be within the protection scope of the present invention.

Claims

1. 2. A compound of formula (I), or a pharma- ceutically acceptable salt, solvate, ester, acid, metabolite or prodrug thereof, for use in treating a cancer or tumor in which M2 macrophages predominate in the tumor microenvironment: 【Chemistry 1】 (Wherein, Y is 【Chemistry 2】 is selected from A is selected from aryl or 6-membered heterocyclyl; n is an integer selected from 1 to 3; R １ is hydrogen, halo, C １～６ Alkyl, C １～６ Haloalkyl, C １～６ selected from alkoxy and cyano; R ２ and R ３ Each independently represents hydrogen, halo, C １～６ Alkyl, C １～６ Haloalkyl, C １～６ alkoxy or R ２ and R ３ together form phenyl or a 5-membered heterocyclyl).

2. 2. The compound of formula (I) for use according to claim 1, or a pharma- ceutically acceptable salt, solvate, ester, acid, metabolite or prodrug thereof, wherein A is selected from phenyl, N-morpholinyl, N-piperidyl or N-piperazinyl.

3. Y, 【Chemistry 3】 When n is 1 and Y is 【Chemistry 4】 3. A compound of formula (I) for use according to claim 1 or 2, wherein, when

4. R １ is selected from hydrogen, fluoro, chloro, methyl, ethyl, propyl, or cyano; R ２ and R ３ are each independently selected from hydrogen, fluoro, chloro, methyl, ethyl, propyl, trifluoromethyl, difluoromethyl, trifluoroethyl, methoxy, ethoxy, propoxy, or R ２ and R ３ or a pharma- ceutically acceptable salt, solvate, ester, acid, metabolite or prodrug thereof, for use according to any one of claims 1 to 3, wherein

5. The compound of formula (I) may be a compound of formula (Ia): 【Chemistry 5】 (In the formula, R １ is selected from hydrogen, fluoro, chloro, and methyl; R ２ and R ３ are each independently selected from hydrogen, fluoro, chloro, methyl, and trifluoromethyl, or R ２ and R ３ (a) taken together to form phenyl; or a pharma- ceutically acceptable salt, solvate, ester, acid, metabolite or prodrug thereof.

6. The compound of formula (I) may be a compound of formula (Ib): 【Chemistry 6】 wherein A is selected from phenyl or N-morpholinyl; R １ is hydrogen, R ２ and R ３ or a pharma- ceutically acceptable salt, solvate, ester, acid, metabolite or prodrug thereof, for use according to any one of claims 1 to 4, wherein each is independently selected from hydrogen, fluoro, chloro, methyl, and trifluoromethyl.

7. The compound of formula (I) may be the compound: 【Table 6】 7. A compound of formula (I) for use according to any one of claims 1 to 6, or a pharma- ceutically acceptable salt, solvate, ester, acid, metabolite or prodrug thereof, selected from:

8. 8. The compound of formula (I) for use according to any one of claims 1 to 7, or a pharmaceutically acceptable salt, solvate, ester, acid, metabolite or prodrug thereof, for use according to any one of claims 1 to 7, wherein the cancer or tumor is selected from one or more of ovarian cancer, cervical cancer, membranous adenocarcinoma, prostate cancer, bladder cancer, lung cancer, thyroid cancer, breast cancer, pancreatic cancer, kidney cancer, gastric cancer, liver cancer, cervical cancer, endometrial cancer, colorectal cancer, nasopharyngeal cancer, esophageal cancer, bile duct cancer, bone metastasis cancer, papillary thyroid cancer, non-small cell lung cancer, small cell lung cancer, colon cancer, solid tumors, brain tumors, lymphoma, glioma, glioblastoma, melanoma, mesothelioma, glioblastoma, osteosarcoma, gastrointestinal stromal tumor, multiple myeloma, bile duct carcinosarcoma, and leukemia.

9. 9. The compound of formula (I), or a pharma- ceutically acceptable salt, solvate, ester, acid, metabolite or prodrug thereof, for use according to claim 8, wherein the lymphoma is selected from non-Hodgkin's lymphoma, B-cell or T-cell lymphoma.

10. 9. The compound of formula (I), or a pharma- ceutically acceptable salt, solvate, ester, acid, metabolite or prodrug thereof, for use according to claim 8, wherein the leukemia is selected from chronic myeloid leukemia or acute myeloid leukemia.

11. 1. A method of treating a cancer or tumor in which M2 macrophages predominate in the tumor microenvironment, comprising administering to a patient a therapeutically effective amount of a compound of formula (I), or a pharma- ceutically acceptable salt, solvate, ester, acid, metabolite, or prodrug thereof: 【Chemistry 7】 (Wherein, Y is 【Chemistry 8】 is selected from A is selected from aryl or 6-membered heterocyclyl; n is an integer selected from 1 to 3; R １ is hydrogen, halo, C １～６ Alkyl, C １～６ Haloalkyl, C １～６ selected from alkoxy and cyano; R ２ and R ３ Each independently represents hydrogen, halo, C １～６ Alkyl, C １～６ Haloalkyl, C １～６ alkoxy or R ２ and R ３ together form a phenyl or a 5-membered heterocyclyl. The method of claim 1, further comprising administering

12. The method of claim 11, wherein A is selected from phenyl, N-morpholinyl, N-piperidyl, or N-piperazinyl.

13. Y, 【Chemistry 9】 When n is 1 and Y is 【Chemistry 10】 13. The method of claim 11 or 12, wherein n is 3 when

14. R １ is selected from hydrogen, fluoro, chloro, methyl, ethyl, propyl, or cyano; R ２ and R ３ are each independently selected from hydrogen, fluoro, chloro, methyl, ethyl, propyl, trifluoromethyl, difluoromethyl, trifluoroethyl, methoxy, ethoxy, propoxy, or R ２ and R ３ The method of any one of claims 11 to 13, wherein combine to form a phenyl or dioxolane.

15. The compound of formula (I) may be a compound of formula (Ia): 【Chemistry 11】 (In the formula, R １ is selected from hydrogen, fluoro, chloro, and methyl; R ２ and R ３ are each independently selected from hydrogen, fluoro, chloro, methyl, and trifluoromethyl, or R ２ and R ３ The method according to any one of claims 11 to 14, wherein said aryl is selected from the group consisting of:

16. The compound of formula (I) may be a compound of formula (Ib): 【Chemistry 12】 wherein A is selected from phenyl or N-morpholinyl; R １ is hydrogen, R ２ and R ３ The method of any one of claims 11 to 14, wherein each is independently selected from hydrogen, fluoro, chloro, methyl, and trifluoromethyl.

17. The compound of formula (I) may be the compound: 【Table 7】 The method according to any one of claims 11 to 16, wherein the method is selected from the group consisting of

18. 18. The method of any one of claims 11 to 17, wherein the cancer or tumor is selected from one or more of ovarian cancer, cervical cancer, membranous adenocarcinoma, prostate cancer, bladder cancer, lung cancer, thyroid cancer, breast cancer, pancreatic cancer, kidney cancer, gastric cancer, liver cancer, cervical cancer, endometrial cancer, colorectal cancer, nasopharyngeal cancer, esophageal cancer, bile duct cancer, bone metastasis cancer, papillary thyroid cancer, non-small cell lung cancer, small cell lung cancer, colon cancer, solid tumors, brain tumors, lymphomas, gliomas, glioblastomas, melanomas, mesothelioma, glioblastomas, osteosarcomas, gastrointestinal stromal tumors, multiple myeloma, bile duct carcinosarcoma, and leukemia.

19. 19. The method of claim 18, wherein the lymphoma is selected from non-Hodgkin's lymphoma, B-cell or T-cell lymphoma.

20. 19. The method of claim 18, wherein the leukemia is selected from chronic myeloid leukemia or acute myeloid leukemia.

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