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WO2014071824A1 - 4-quinazolinamine heterocyclic compound and use thereof - Google Patents

4-quinazolinamine heterocyclic compound and use thereof Download PDF

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Publication number
WO2014071824A1
WO2014071824A1 PCT/CN2013/086514 CN2013086514W WO2014071824A1 WO 2014071824 A1 WO2014071824 A1 WO 2014071824A1 CN 2013086514 W CN2013086514 W CN 2013086514W WO 2014071824 A1 WO2014071824 A1 WO 2014071824A1
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cxm
fluoro
benzyl
ylamino
oxazol
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PCT/CN2013/086514
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French (fr)
Chinese (zh)
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唐田
于琳
冯汉林
严启新
王兵
王志国
朱丹
陈红
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深圳海王药业有限公司
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Publication of WO2014071824A1 publication Critical patent/WO2014071824A1/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/14—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing three or more hetero rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links

Definitions

  • the present invention relates to a therapeutically active 4-aminoquinazoline heterocyclic compound, a process for the preparation thereof, a pharmaceutical composition containing these compounds, and the use of said compound or pharmaceutical composition for the preparation of an antitumor drug.
  • Cancer/malignant tumors are one of the major threats to human health. Most tumors are related to environmental factors, and more than 5 million people worldwide die from cancer every year. Although there are some treatments, such as surgery, radiotherapy, chemotherapy, etc., the patient can be cured, but the cure rate is not high. At present, the use of chemical drugs for prevention and treatment is one of the most effective methods for subduing tumors.
  • EGFR epidermal growth factor receptor
  • EGFR-TK inhibitors both as a promising antitumor drug, exhibit significant antitumor effects, either alone or in combination with assimilation.
  • Anti-tumor drugs such as Gefitinib, Erlotinib, Lapatinib, etc., which have been marketed, are the first generation of EGFR-TK inhibitors.
  • Clinical practice has found that tumor patients are less sensitive to first-generation EGFR-TK inhibitors, and approximately 10% of North American patients and approximately 20% of Asian patients respond to them.
  • second-generation EGFR inhibitors have entered clinical trials.
  • One advantage of second-generation EGFR inhibitors is that they are irreversibly covalently bound to the target, so that the drug acts on the entire pathway of the epidermal growth factor signal transduction pathway and increases the blocking efficiency of the drug. This irreversible combination kills tumor cells that are resistant to first-generation EGFR inhibitors.
  • Another advantage is multi-targeting. Tumor cell signaling is diverse, and second-generation EGFR inhibitors, in addition to blocking the EGFR signaling pathway, block the ErbB family of signal transduction pathways such as HER-2. The results of the second-generation EGFR inhibitors in clinical stages I and II are attractive, especially BIBW2992, which is undergoing Phase II clinical trials.
  • WO0250043, US20050085495, US20090306378, and the like describe compounds similar to BIBW2992, which have important pharmacological properties, particularly inhibitory effects on signal transmission mediated by tyrosine kinases and through epidermal growth factor receptors ( The signal transmission mediated by EGF-R) has an inhibitory effect. Therefore, these compounds are particularly useful for the treatment of neoplastic diseases, lung and respiratory diseases, and gastrointestinal, bile duct and gallbladder diseases.
  • the present inventors designed and synthesized a novel 4-aminoquinazoline compound based on the development of a multi-target tyrosine kinase inhibitor, which inhibits tumor angiogenesis, irreversibly inhibits EGFR and related HER-2, and inhibits cell mitosis. Various routes are thus used to treat tumor diseases. Summary of the invention
  • Another object of the present invention is to provide a pharmaceutical composition
  • a pharmaceutical composition comprising the above compound and a pharmaceutically acceptable carrier.
  • Still another object of the present invention is to provide use of the above compound or pharmaceutical composition for the preparation of an antitumor drug, preferably a disease mediated by a protein tyrosine kinase, such as breast cancer, non-small cell lung cancer, ovarian cancer , stomach cancer, pancreatic cancer, etc.
  • an antitumor drug preferably a disease mediated by a protein tyrosine kinase, such as breast cancer, non-small cell lung cancer, ovarian cancer , stomach cancer, pancreatic cancer, etc.
  • R 5 represents hydrazine, hydrazine, hydrazine-dimethylaminomethyl, hydrazine, hydrazine-diethylaminomethyl or hydrazine, fluorenyl-dipropylaminomethyl;
  • R 3 represents hydrogen or a C 1-4 yard group.
  • Ri, R 2 independently represents the following structure:
  • X represents 0 or S
  • n represents an integer from 0 to 4, such as 0, 1, 2, 3 or 4
  • R 4 represents hydrogen, CM thiol, C M methoxy, 3-tetrahydrofuranyl, morpholinyl, 4-tetrahydropyranyl, 2-pyrrolyl, oxazolinyl, thiazolyl or thienyl; or
  • Ri, R 2 independently represents the following structure:
  • R 5 represents hydrazine, hydrazine, hydrazine-dimethylaminomethyl, hydrazine, hydrazine-diethylaminomethyl or hydrazine, hydrazine-dipropylaminomethyl; preferred compounds of the invention are selected from the group consisting of:
  • the compounds of the present invention may be in the form of tautomers, stereoisomers or pharmaceutically acceptable salts thereof.
  • Halogen means fluoro, chloro, bromo and iodo, preferably fluoro, chloro or bromo.
  • C M thiol means methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl and tert-butyl; preferably methyl, ethyl, propyl, isopropyl or butyl More preferably a methyl group.
  • C M methoxy methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy and tert-butoxy; preferably methoxy, Ethoxy, propoxy, isopropoxy or butoxy; more preferably methoxy.
  • the "ester group” nail acrylate group, acetate group, propionate group, butyrate group or the like is preferably an acetate group.
  • “Sulfolamido” is a sulfonamide group, an ethylsulfonylamino group, a propylsulfonylamino group, an isopropylsulfonylamino group or the like; preferably a methanesulfonylamino group.
  • the "phenyl group” may be a phenyl group substituted with a mercapto group, an anthracenyloxy group, a nonyloxymethyl group, an ester group, a sulfonate group, a hydroxyl group or the like, preferably a decyloxy group, a decyloxymethyl group, an ester group, or a hydroxyl group. Substituted phenyl.
  • “Furanyl” includes 2-furyl and 3-furyl.
  • “Tetrahydrofuranyl” includes 2-tetrahydrofuranyl and 3-tetrahydrofuranyl. Also included are furyl and tetrahydrofuranyl groups substituted with groups such as decyl, decyloxy, decyloxymethyl, ester, sulfonate, hydroxy, and the like.
  • “Pyryl” includes 2-pyranyl, 3-pyranyl and 4-pyranyl.
  • “Tetrahydropyranyl” includes 2-tetrahydropyranyl, 3-tetrahydropyranyl and 4-tetrahydropyranyl. Also included are pyranyl and tetrahydropyranyl groups substituted with a group such as a mercapto group, a decyloxy group, a decyloxymethyl group, an ester group, a sulfonate group, a hydroxyl group or the like.
  • Thiophenyl includes 2-thienyl and 3-thienyl. Also included are thienyl groups substituted with mercapto, decyloxy, decyloxymethyl, ester, sulfonate, hydroxy, and the like.
  • Steps include cis and trans isomers, enantiomers and conformational isomers.
  • “Pharmaceutically acceptable salt” refers to a relatively non-toxic acid addition salt of a compound of formula (I) of the present invention with an organic acid or inorganic. These salts can be prepared in situ during the final separation and purification of the compound, or by reacting the compound in its free base form with a suitable inorganic or organic acid.
  • the inorganic or organic acid is selected from the group consisting of hydrobromic acid, hydrochloric acid, sulfuric acid, sulfurous acid, acetic acid, oxalic acid, valeric acid, oleic acid, palmitic acid, stearic acid, lauric acid, boric acid, benzoic acid, lactic acid, phosphoric acid, toluene.
  • hydrobromic acid hydrochloric acid
  • sulfuric acid sulfurous acid
  • acetic acid oxalic acid
  • valeric acid oxalic acid
  • valeric acid oleic acid
  • palmitic acid stearic acid
  • lauric acid boric acid
  • benzoic acid lactic acid, phosphoric acid, toluene.
  • citric acid maleic acid, fumaric acid, succinic acid, tartaric acid, benzoic acid, methanesulfonic acid, gluconic acid, lactobionic acid and lauryl sulfonic acid.
  • a pharmaceutical composition comprising a compound of the invention and a pharmaceutically acceptable carrier.
  • the compounds of the present invention may be administered alone or in combination with each other or in combination with other pharmaceutically acceptable active compounds.
  • the compound can be administered to a human or animal in need thereof by oral, rectal, parenteral, or topical administration in the form of a pharmaceutical composition.
  • compositions are usually prepared by mixing the active compound with one or more pharmaceutically acceptable carriers according to conventional methods.
  • the pharmaceutically acceptable carrier refers to a conventional inert carrier in the pharmaceutical art.
  • solid dosage forms for oral administration include capsules, tablets, pills, powders, granules and the like.
  • the pharmaceutical composition may comprise one or more of the following carriers: diluents such as starch, lactose, sucrose, glucose, mannitol or silicic acid; binders such as hydroxymethylcellulose, alginate , gelatin, polyvinylpyrrolidone, sucrose and gum arabic; dispersing agents such as polyethylene glycol, polyvinylpyrrolidone; humectants such as glycerin; disintegrating agents such as agar, calcium carbonate, potato starch or tapioca Starch, alginic acid, certain complex silicates and sodium carbonate; slow solvents such as paraffin; absorption accelerators, such as quaternary amine compounds; wetting agents, such as cetyl alcohol and glyceryl monostearate; adsorbents, such as kaolin Lubricants, such as tal
  • Liquid dosage forms for oral administration include emulsions, solutions, suspensions, dragees or elixirs.
  • the pharmaceutical composition may comprise one or more of the following carriers: diluents such as water or other solvents, solubilizers and emulsifiers such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1, 3-butanediol, dimethylformamide and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances; suspending agents, such as ethoxylated iso-eighteen Sterols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar.
  • adjuvants such as wetting agents, emulsifying agents, suspending agents, sweetening agents, flavoring agents and perfumes may be included.
  • Dosage forms for parenteral administration include intravenous, subcutaneous, intramuscular injection preparations.
  • the pharmaceutical compositions may comprise physiologically acceptable sterile aqueous or nonaqueous solutions, dispersions, suspensions A liquid or emulsion, and a sterile powder for reconstitution into a sterile injectable solution or dispersion.
  • Suitable aqueous and nonaqueous vehicles, diluents, and solvents include water, ethanol, polyols, and suitable mixtures thereof.
  • Dosage forms for topical administration include ointments, powders, propellants, drops, and inhalants.
  • the active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants which may be required if necessary.
  • a compound or a pharmaceutical composition of the present invention for the preparation of an antitumor drug, preferably a disease mediated by a protein tyrosine kinase, such as breast cancer, non-small cell lung cancer , ovarian cancer, stomach cancer, pancreatic cancer, etc.
  • an antitumor drug preferably a disease mediated by a protein tyrosine kinase, such as breast cancer, non-small cell lung cancer , ovarian cancer, stomach cancer, pancreatic cancer, etc.
  • the compound 12 (472.1 g, 1 mol) was dissolved in methanol (5 L), concentrated aqueous ammonia (0.5 L) was added dropwise, and the mixture was reacted at room temperature for 5 h, the reaction liquid was filtered, and the filtrate was concentrated. The concentrate was combined with the filter cake and recrystallized from methanol.
  • the preparation method was similar to that of Example 13, except that sodium methoxide was replaced with sodium ethoxide.
  • the preparation method was similar to that of Examples 4 and 5, except that the starting material 4-chloro-6-nitro-quinazolin-7-yl-acetate was replaced with 4-chloro-7-nitro-quinazoline. -6-yl-acetate.
  • the title product was obtained from the title compound C.M.
  • the preparation method was similar to that of Example 7, except that 4-chloro-7-nitro-quinazolin-6-yl-acetate was used instead of 4-chloro-6-nitro-quinazolin-7-yl-B.
  • the acid ester serves as a starting material.
  • the title product was obtained as the title compound: EtOAc (EtOAc): m.
  • the preparation method was similar to that in Example 8, except that 4-chloro-7-nitro-quinazolin-6-yl-acetate was used instead of 4-chloro-6-nitro-quinazolin-7-yl-B.
  • the acid ester serves as a starting material.
  • the title product was the title compound C.M.s.
  • the preparation method was similar to that of Example 9, except that 4-chloro-7-nitro-quinazolin-6-yl-acetate was used instead of 4-chloro-6-nitro-quinazolin-7-yl-B.
  • the acid ester serves as a starting material.
  • the preparation method was similar to that in Example 10 except that 4-chloro-7-nitro-quinazolin-6-yl-acetate was used instead of 4-chloro-6-nitro-quinazolin-7-yl- Acetate is used as a starting material.
  • the preparation method was similar to that in Example 15, except that 4-chloro-7-nitro-quinazolin-6-yl-acetate was used instead of 4-chloro-6-nitro-quinazolin-7-yl-B.
  • the acid ester serves as a starting material.
  • the title product was obtained as the title compound C.M.s.
  • the preparation method was similar to that in Example 16, except that 4-chloro-7-nitro-quinazolin-6-yl-acetate was used instead of 4-chloro-6-nitro-quinazolin-7-yl-B.
  • the acid ester serves as a starting material.
  • TK-biotin substrate Cat. 62TKOPEC, Cisbio
  • ATP Cat. PV3227, Invitrogen
  • Enzyme FGFRl (Cat. PV3146, Invitrogen), PDGFR beta (Cat. P3082, Invitrogen) , EGFR (Cat. PV3872, Invitrogen), KDR (Cat. K2643, Sigma), HER2 (Cat. PV3366, Invitrogen); Streptavidin-XL665 (Cat.
  • 62TKOPEC Cisbio
  • TK antibody europium cryptate Cat.62TKOPEC, Cisbio
  • HTRF kinEASE TK kit Cat.62TKOPEC, Cisbio
  • ENVISION detector Perkinelmer (Cat. 2104-0010, PerkinElmer).
  • kinase reagents Five kinase reagents, FGFR1, PDGF, KDR, EGFR and HER2, were formulated as shown in Table 1.
  • Enzyme reaction time 25 min 20 min 20 min 30 min 30 min
  • the concentration of Streptavidin-XL665 is 58.13 ⁇ 125 ⁇ 25 ⁇ 20.6 ⁇ 37.5 ⁇ in the total reaction.
  • TK Antibody-Cryptate concentration final concentration (20 ⁇ ) 1 100 dilution 1: 100 dilution 1: 100 dilution 1: 100 dilution 1: 100 dilution 1: 100 dilution l xFGFRl
  • Enzymatic Buffer: lmL 1 x Kinase Buffer contains 200 ⁇ L 5 x Enzyme buffer, 5 ⁇ 1M MgCl 2 , ⁇ 1M DTT, 794 dd3 ⁇ 40.
  • 1 xPDGFR Enzymatic Buffer lmL 1 x Kinase Buffer contains 200 ⁇ L 5> ⁇ Enzyme buffer, 5 ⁇ 1M MgCl 2 , ⁇ 1M DTT, 20 ⁇ SEB, 1 1M MnCl 2 , 773 ⁇ ddH 2 0.
  • Lx KDR Enzymatic Buffer contains 200 ⁇ L 5 x Enzyme buffer, 5 ⁇ 1M MgCl 2 , ⁇ 1M DTT, 1 ⁇ 1M MnCl 2 , 793 ⁇ dd3 ⁇ 40.
  • Lx EGFR Enzymatic Buffer contains 200 ⁇ L 5> ⁇ Enzyme buffer, 5 ⁇ 1M MgCl 2 , ⁇ 1M DTT, 1 ⁇ 1M MnCl 2 , 793 ⁇ dd3 ⁇ 40.
  • l xHER2 Enzymatic Buffer contains 200 ⁇ L 5 x Enzyme buffer, 5 1M MgCl 2 , ⁇ 1M DTT, 3.2 ⁇ SEB, 1 1M MnCl 2 , 789.8 ddH 2 0.
  • Substrate-TK and ATP were diluted to 5 times the reaction concentration with 1 x Kinase Buffer.
  • 5xEnzyme working solution The concentration of the reagents used in the five kinase concentration optimization screening is shown in Table 1. Five enzymes of 5 enzymes were prepared using l xkinase buffer.
  • TK-Antibody-cryptate working solution Dilute TK- Antibody-Cryptate 100 times as a working solution with Detection Buffer.
  • the enzyme was added to the room temperature after the enzyme was added to the room temperature.
  • a) First prepare a 2.5% DMSO solution using the IX kinase buffers of the five enzymes configured (the DMSO concentration will affect the reaction, control the final concentration of DMSO to 1%), and then use the five enzymes corresponding to 2.5.
  • the test compound was diluted with % DMSO solution, and the final concentration of each analyte was 2 ⁇ and 0.2 ⁇ .
  • Streptavidin-XL665 solution add the diluted TK antibody europium cryptate test solution immediately after mixing.
  • the plate was mixed and reacted at room temperature for 1 h, and the fluorescence signal (320 nm stimulation, 665 nm, 615 nm emission) was detected by an ENVISION detector.
  • the inhibition rate of each well was calculated from the fully active wells and the background signal wells, and the duplicate wells were averaged, and the half-inhibitory activity (IC50) of each test compound was fitted using a professional paint analysis software PRISM 5.0.
  • Emissivity (ER) 665 nm transmit signal / 615 nm transmit signal
  • Inhibition rate (ER earn - ER sample) / (ER positive - ER negative) * 100%
  • the experimental sample flow chart is as follows:
  • Results and analysis Tables 2 - 6 show the inhibition rates of the FGFRl, PDGFR, KDR, EGFR and HER2 enzymes of the compounds of the invention at concentrations of 2 ⁇ and 0.2 ⁇ , respectively.
  • the compound of the invention CXM-30, 33, 02, 05, 12, 09, 24; positive control BIBW-2992, lapatinib: purchased from Hangzhou Rongda Pharmaceutical Chemical Co., Ltd.; non-small cell cancer cell NCI-H322M, Prostate cancer cell line DU145: purchased from American Type Culture Collection ATCC; liver cancer cell Hep3B, breast cancer cell MDA-MB-231, ovarian cancer cell SK-OV-3, hypopharyngeal carcinoma human pharyngeal carcinoma cell FaDu, malignant collagen Cell tumor cell U-87MG: purchased from Shanghai Institute of Cell Biology, Chinese Academy of Sciences; CCK-8 test kit: purchased from Cat# CK04-13, Dojindo.
  • Each cell line was cultured with the corresponding complete medium below, and the cells were grown in a incubator at 37 ° C, 100% relative humidity, 5 % 0 2 , and cells in logarithmic growth phase were selected for experimental testing.
  • experiment a) Collect logarithmic growth phase cells, count, resuspend the cells with complete medium, adjust the cell concentration to the appropriate concentration (determined according to the cell density optimization test results), inoculate 96-well plates, and add ⁇ cell suspension to each well. The cells were incubated for 24 h at 37 ° C, 100% relative humidity, 5% C0 2 incubator.
  • test compound Dilute the test compound to 5 ⁇ & 0.5 ⁇ with medium, and add the cells to 25 ⁇ l/well to a final concentration of 1 ⁇ to 0.1 ⁇ .
  • Tumor cell growth inhibition rate % [( 0 8 8) / ( (;- ): ⁇ 100%
  • the compound of the present invention and the positive control were separately detected by the CCK-8 test kit for different cell lines (non-small cell cancer cell NCI-H322M, liver cancer cell Hep3B, breast cancer cell MDA-MB-231, prostate cancer).
  • non-small cell cancer cell NCI-H322M liver cancer cell Hep3B
  • breast cancer cell MDA-MB-231 prostate cancer.
  • the final concentration of the compound diluted with DMSO was from ⁇ to 0 ⁇ , including 10 ⁇ , 2 ⁇ , 0 ⁇ 4 ⁇ , 0 ⁇ 08 ⁇ , 0 ⁇ 016 ⁇ , 0 ⁇ 0032 ⁇ , 0 ⁇ 00064 ⁇ , 0.000128 ⁇ , and 0 ⁇ . 3.
  • Reagents Compounds of the invention: CXM-02 CXM-30 CXM-33 CXM- 12 CXM-09 CXM-24; Positive control: Lapatinib (product of Hangzhou Rongda Pharmaceutical Chemical Co., Ltd.), BIBW -2992 (Hangzhou Rongda Pharmaceutical Chemical Co., Ltd. Product Company);
  • the concentration of the test substance in the rat blood plasma was determined by liquid chromatography-mass spectrometry-mass spectrometry (LC/MS/MS). Based on the obtained blood drug concentration data, the pharmacokinetic processing software WinNonlin 5.2 non-compartment model was used to calculate the relevant pharmacokinetic parameters.

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Abstract

A compound of (I), preparation methods therefor, and the use of the compound for preparing antineoplastic agents.

Description

4-氨基喹唑啉杂环化合物及其用途  4-aminoquinazoline heterocyclic compound and use thereof
技术领域 Technical field
本发明涉及有治疗活性的 4-氨基喹唑啉杂环化合物及其制备方法，含有这 些化合物的药物组合物，以及所述化合物或药物组合物在制备抗肿瘤药物中的 用途。 技术背景  The present invention relates to a therapeutically active 4-aminoquinazoline heterocyclic compound, a process for the preparation thereof, a pharmaceutical composition containing these compounds, and the use of said compound or pharmaceutical composition for the preparation of an antitumor drug. technical background
癌症 /恶性肿瘤是人类健康的主要威胁之一， 大多数肿瘤都与外界环境因 素相关， 全世界每年有 500万人以上死于癌症。 虽然现在已有一些治疗办法， 如外科手术、 放疗、 化疗等可使患者治愈， 但治愈率不高。 目前， 使用化学药 物预防及治疗是制服肿瘤的最有效方法之一。  Cancer/malignant tumors are one of the major threats to human health. Most tumors are related to environmental factors, and more than 5 million people worldwide die from cancer every year. Although there are some treatments, such as surgery, radiotherapy, chemotherapy, etc., the patient can be cured, but the cure rate is not high. At present, the use of chemical drugs for prevention and treatment is one of the most effective methods for subduing tumors.
表皮生长因子受体 （EGFR) 信号传导通路在肿瘤细胞的生成、 增殖和生 存方面起着重要的作用。 因此， EGFR—直是抗肿瘤药物的一个重要的靶标。 目前为止， 体内外的试验和临床研究显示， 作为一种有前景的抗肿瘤药物， EGFR-TK抑制剂无论是单独应用还是同化放疗联合都表现出显著的抗肿瘤作 用。 已经上市的抗肿瘤药物如吉非替尼 （Gefitinib)、 埃罗替尼 （Erlotinib)、 拉帕替尼 （Lapatinib) 等是第一代 EGFR-TK抑制剂。 临床实践中发现， 肿瘤 患者对第一代 EGFR-TK抑制剂的敏感性较低，约 10%的北美患者和约 20%的 亚洲患者对它们有响应。 目前， 第二代 EGFR抑制剂已进入临床试验阶段。第 二代 EGFR抑制剂的一个优点是它们与靶标不可逆地共价结合，这样，药物就 能作用到表皮生长因子信号转导通路的整个环节， 并提高药物的阻断效率。这 种不可逆性结合能够杀死那些对第一代 EGFR抑制剂耐药的肿瘤细胞。另一个 优点是多靶性。肿瘤细胞的信号传导具有多样性，第二代 EGFR抑制剂除了阻 止 EGFR信号转导通路以外，还可以阻止像 HER-2这样的 ErbB家族的信号转 导通路。第二代 EGFR抑制剂在临床 I和 II期中的试验结果是有吸引力的，特 别是正在 II期临床试验的 BIBW2992。  The epidermal growth factor receptor (EGFR) signaling pathway plays an important role in the generation, proliferation and survival of tumor cells. Therefore, EGFR is an important target for anti-tumor drugs. To date, in vitro and in vivo trials and clinical studies have shown that EGFR-TK inhibitors, both as a promising antitumor drug, exhibit significant antitumor effects, either alone or in combination with assimilation. Anti-tumor drugs such as Gefitinib, Erlotinib, Lapatinib, etc., which have been marketed, are the first generation of EGFR-TK inhibitors. Clinical practice has found that tumor patients are less sensitive to first-generation EGFR-TK inhibitors, and approximately 10% of North American patients and approximately 20% of Asian patients respond to them. Currently, second-generation EGFR inhibitors have entered clinical trials. One advantage of second-generation EGFR inhibitors is that they are irreversibly covalently bound to the target, so that the drug acts on the entire pathway of the epidermal growth factor signal transduction pathway and increases the blocking efficiency of the drug. This irreversible combination kills tumor cells that are resistant to first-generation EGFR inhibitors. Another advantage is multi-targeting. Tumor cell signaling is diverse, and second-generation EGFR inhibitors, in addition to blocking the EGFR signaling pathway, block the ErbB family of signal transduction pathways such as HER-2. The results of the second-generation EGFR inhibitors in clinical stages I and II are attractive, especially BIBW2992, which is undergoing Phase II clinical trials.
随着作用机制的进一步阐明， 将会有更多的此类药物进入临床， 为肿瘤的 治疗提供更为广阔的发展空间。 例如， WO0250043、 US20050085495、 US20090306378等专利申请描述了类似 BIBW2992 的化合物， 它们具有重要 药理学性质，尤其是对通过酪氨酸激酶所介导的讯号传递具有抑制效果以及对 通过表皮生长因子受体（EGF-R)所介导的讯号传递具有抑制效果。 因此， 这 些化合物特别适用于治疗肿瘤疾病、肺部与呼吸道疾病以及胃肠道、胆管与胆 囊疾病。 本发明人基于多靶点酪氨酸激酶抑制剂的开发， 设计并合成新的 4-氨基喹 唑啉化合物， 其通过抑制肿瘤血管生成， 不可逆地抑制 EGFR和相关 HER-2 以及抑制细胞有丝分裂的各种途径， 从而用于治疗肿瘤疾病。 发明内容 With the further clarification of the mechanism of action, more such drugs will enter the clinic, providing a broader space for the development of tumors. For example, WO0250043, US20050085495, US20090306378, and the like, describe compounds similar to BIBW2992, which have important pharmacological properties, particularly inhibitory effects on signal transmission mediated by tyrosine kinases and through epidermal growth factor receptors ( The signal transmission mediated by EGF-R) has an inhibitory effect. Therefore, these compounds are particularly useful for the treatment of neoplastic diseases, lung and respiratory diseases, and gastrointestinal, bile duct and gallbladder diseases. The present inventors designed and synthesized a novel 4-aminoquinazoline compound based on the development of a multi-target tyrosine kinase inhibitor, which inhibits tumor angiogenesis, irreversibly inhibits EGFR and related HER-2, and inhibits cell mitosis. Various routes are thus used to treat tumor diseases. Summary of the invention
本发明的一个目的是提供新的 4-氨基喹唑啉杂环化合物。  It is an object of the present invention to provide novel 4-aminoquinazoline heterocyclic compounds.
本发明的另一个目的是提供含有上述化合物以及药学上可接受的载体的 药物组合物。  Another object of the present invention is to provide a pharmaceutical composition comprising the above compound and a pharmaceutically acceptable carrier.
本发明的再一个目的是提供上述化合物或药物组合物在制备抗肿瘤药物 中的用途， 所述肿瘤优选为由蛋白络氨酸激酶介导的疾病， 例如乳腺癌、 非小 细胞肺癌、 卵巢癌、 胃癌、 胰腺癌等。  Still another object of the present invention is to provide use of the above compound or pharmaceutical composition for the preparation of an antitumor drug, preferably a disease mediated by a protein tyrosine kinase, such as breast cancer, non-small cell lung cancer, ovarian cancer , stomach cancer, pancreatic cancer, etc.
根据本发明的一 面， 提供式 （I) 所示的化合物：  According to one aspect of the invention, a compound of formula (I) is provided:
Figure imgf000004_0001
Figure imgf000004_0001
其中， 和 独立地表示氢、 卤素、 CM垸基、 卤素取代的 CM垸基、 CM垸氧基、 卤素取代的 CM垸氧基、 腈基、 酯基、 硝基、 氨基、 酰胺基、 磺 酰胺基、 苯基或 CH3S02CH2CH2NHCH2-Ar; 其中 Ar表示苯基、 呋喃基、 噻吩 基、 吡咯基和噻唑基， 并且 Ar可被一个或两个选自卤素、 d_4垸基和 CM垸 氧基的基团取代； 或者 Wherein, and independently represent hydrogen, halogen, C M fluorenyl, halogen substituted CM fluorenyl, CM methoxy, halogen substituted CM methoxy, cyano, ester, nitro, amino, amide, sulfonate Amido, phenyl or CH 3 S0 2 CH 2 CH 2 NHCH 2 -Ar ; wherein Ar represents phenyl, furyl, thienyl, pyrrolyl and thiazolyl, and Ar may be selected from one or two selected from halogen, d_ Substituted with a 4 -mercapto group and a CM methoxy group; or
Ri, R2独立地表示： Ri, R 2 stands independently:
X-(CH2)n-R4 X-(CH 2 ) n -R 4
其中 X表示 N、 0或 S; n表示 0到 6的整数； R4表示氢、 CM垸基、 卤 素取代的 (^_4垸基、 CM垸氧基、 卤素取代的 CM垸氧基、 呋喃基、 四氢呋喃 基、 吗啉基、 吡喃基、 四氢吡喃基、 吡咯基、 吡啶基、 噁唑啉基、 噻唑基或噻 吩基； 或者 Wherein X represents N, 0 or S; n represents an integer from 0 to 6; R 4 represents hydrogen, C M fluorenyl, halogen substituted (^_4 fluorenyl, CM methoxy, halogen-substituted CM methoxy, furan) , tetrahydrofuranyl, morpholinyl, pyranyl, tetrahydropyranyl, pyrrolyl, pyridyl, oxazolinyl, thiazolyl or thienyl;
Ri、 R2独立地表示：
Figure imgf000004_0002
Ri, R 2 stand independently:
Figure imgf000004_0002
其中 R5表示 Η、 Ν, Ν-二甲基胺基甲基、 Ν, Ν-二乙基胺基甲基或 Ν, Ν-二 丙基胺基甲基； R3表示氢或 C1-4院基。 Wherein R 5 represents hydrazine, hydrazine, hydrazine-dimethylaminomethyl, hydrazine, hydrazine-diethylaminomethyl or hydrazine, fluorenyl-dipropylaminomethyl; R 3 represents hydrogen or a C 1-4 yard group.
在优选的实施方式中， 式 （I) 所示的化合 h 其中：  In a preferred embodiment, the compound h represented by formula (I) wherein:
1^和 独立地表示氢、 CM垸氧基、 硝基. 氨基、 酰胺基； 或者  1^ and independently represent hydrogen, CM methoxy, nitro. amino, amide; or
Ri, R2独立地表示以下结构： Ri, R 2 independently represents the following structure:
X-(CH2)n-R4 X-(CH 2 ) n -R 4
其中 X表示 0或 S; n表示 0到 4的整数， 例如 0、 1、 2、 3或 4; R4表 示氢、 CM垸基、 CM垸氧基、 3-四氢呋喃基、 吗啉基、 4-四氢吡喃基、 2-吡咯 基、 噁唑啉基、 噻唑基或噻吩基； 或者， Wherein X represents 0 or S; n represents an integer from 0 to 4, such as 0, 1, 2, 3 or 4; R 4 represents hydrogen, CM thiol, C M methoxy, 3-tetrahydrofuranyl, morpholinyl, 4-tetrahydropyranyl, 2-pyrrolyl, oxazolinyl, thiazolyl or thienyl; or
Ri, R2独立地表示以下结构：
Figure imgf000005_0001
Ri, R 2 independently represents the following structure:
Figure imgf000005_0001
其中 R5表示 Η、 Ν,Ν-二甲基胺基甲基、 Ν,Ν-二乙基胺基甲基或 Ν,Ν-二 丙基胺基甲基； 本发明优选的化合物选自： Wherein R 5 represents hydrazine, hydrazine, hydrazine-dimethylaminomethyl, hydrazine, hydrazine-diethylaminomethyl or hydrazine, hydrazine-dipropylaminomethyl; preferred compounds of the invention are selected from the group consisting of:
N-{4-[l-(3-氟-苄基） 吲唑 -5-基胺基] -喹唑啉 -6-基 丙烯酰胺 N-{4-[l-(3-Fluoro-benzyl)carbazole-5-ylamino]-quinazoline-6-yl acrylamide
(CXM-01); (CXM-01);
( - {4-[1-(3-氟-苄基) 吲唑 -5-基胺基 ]-7-甲氧基-喹唑啉 -6-基}-4- (二 甲基氨基:)丁 -2-烯酰胺 (CXM-02);  (-{4-[1-(3-Fluoro-benzyl)oxazol-5-ylamino]-7-methoxy-quinazolin-6-yl}-4-(dimethylamino:) But-2-enamide (CXM-02);
( - {4-[1-(3-氟-苄基) 吲唑 -5-基胺基 ]-7-乙氧基-喹唑啉 -6-基}-4- (二 甲基胺基) -丁 -2-烯酰胺 (CXM-03);  (-{4-[1-(3-Fluoro-benzyl)oxazol-5-ylamino]-7-ethoxy-quinazolin-6-yl}-4-(dimethylamino) -but-2-enamide (CXM-03);
7-(2-甲氧基乙氧基) -N-[l-(3-氟-苄基) 吲唑 -5-基] -6-硝基-喹唑啉 -4-胺 (CXM-04);  7-(2-methoxyethoxy)-N-[l-(3-fluoro-benzyl)oxazol-5-yl]-6-nitro-quinazolin-4-amine (CXM-04 );
( -Ν-{4-[1-(3-氟-苄基) 吲唑 -5-基胺基 ]-7-(3-甲氧基乙氧基) -喹唑啉 -6- 基}—4- (二甲基胺基) -丁 -2-烯酰胺 （CXM-05); (- Ν - {4- [1- (3- fluoro-benzyl) - amino-indazol-5-yl] -7- (3-methoxy-ethoxy) - quinazolin-6-yl} - 4-(Dimethylamino)-but-2-enamide (CXM-05);
{4-[1-(3-氟-苄基) 吲唑 -5-基胺基 ]-7-(2-甲氧乙氧基) -喹唑啉 -6-基 丙烯酰胺 (CXM-06);  {4-[1-(3-Fluoro-benzyl)oxazol-5-ylamino]-7-(2-methoxyethoxy)-quinazolin-6-ylacrylamide (CXM-06) ;
N-{4-[l-(3-氟-苄基) 吲唑 -5-基胺基 ]-7-[3-(4-吗啉代)丙氧基] -喹唑啉 -6- 基} -丙烯酰胺 （CXM-07);  N-{4-[1-(3-Fluoro-benzyl)oxazol-5-ylamino]-7-[3-(4-morpholino)propoxy]-quinazoline-6-yl } - acrylamide (CXM-07);
( - {4-[1-(3-氟-苄基) -1H-吲唑 -5-基胺基 ]-7-[3-(4-吗啉代)丙氧基] -喹唑 啉 -6-基}-4- (二甲基胺基) -丁 -2-烯酰胺 （CXM-08); ( -{4-[1-(3-Fluoro-benzyl)-1H-indazol-5-ylamino]-7-[3-(4-morpholino)propoxy]-quinazoline- 6-yl}-4-(dimethylamino)-but-2-enamide (CXM-08);
( )-N-{7-[0¾-四氢呋喃 -3-基氧基 ]-4-[1-(3-氟-苄基) 吲唑 -5-基胺基] -喹 唑啉 -6-基}-4-二甲基胺基-丁 -2-烯酰胺 (CXM-09);  ( )-N-{7-[03⁄4-tetrahydrofuran-3-yloxy]-4-[1-(3-fluoro-benzyl)oxazol-5-ylamino]-quinazolin-6-yl }-4-dimethylamino-but-2-enamide (CXM-09);
( - {4-[1-(3-氟-苄基) 吲唑 -5-基胺基 ]-7- (腈基) -喹唑啉 -6-基}-4-二甲 基胺基-丁 -2-烯酰胺 (CXM-10); ( -N-{7- [(四氢 -2H-吡喃 -4-基) -甲硫基] -4-[l-(3-氟-苄基) 吲唑 -5-基胺 基] -喹唑啉 -6-基}-4-二甲基胺基-丁 -2-烯酰胺 (CXM-11); ( -{4-[1-(3-Fluoro-benzyl)oxazol-5-ylamino]-7-(nitrile)-quinazolin-6-yl}-4-dimethylamino- But-2-enamide (CXM-10); (-N-{7-[(tetrahydro-2H-pyran-4-yl)-methylthio]-4-[l-(3-fluoro-benzyl)oxazol-5-ylamino] - Quinazoline- 6 -yl}-4-dimethylamino-but-2-enamide (CXM-11);
- {4-[1-(3-氟-苄基) 吲唑 -5-基胺基] -喹唑啉 -6-基}-4-二甲基胺基- 丁 -2-烯酰胺 (CXM-12); - {4-[1-(3-Fluoro-benzyl)oxazol-5-ylamino]-quinazolin-6-yl}-4-dimethylamino-but-2-enamide (CXM -12);
{4-[1-(3-氟-苄基：) 吲唑 -5-基胺基 ]-7-甲氧基-喹唑啉 -6-基} -丙烯酰胺 {4-[1-(3-Fluoro-benzyl:)carbazole-5-ylamino]-7-methoxy-quinazoline-6-yl}-acrylamide
(CXM-13); (CXM-13);
N-{4-[l-(3-氟-苄基) 吲唑 -5-基胺基 ]-7-乙氧基-喹唑啉 -6-基 丙烯酰胺 (CXM-14);  N-{4-[1-(3-Fluoro-benzyl)carbazole-5-ylamino]-7-ethoxy-quinazoline-6-yl acrylamide (CXM-14);
N-{7-[0¾-四氢呋喃 -3-基氧基 ]-4-[1-(3-氟-苄基) 吲唑 -5-基胺基] -喹唑啉 -6-基} -丙烯酰胺 (CXM-15);  N-{7-[03⁄4-tetrahydrofuran-3-yloxy]-4-[1-(3-fluoro-benzyl)oxazol-5-ylamino]-quinazolin-6-yl}-propene Amide (CXM-15);
N-{4-[l-(3-氟-苄基) 吲唑 -5-基胺基 ]-7-腈基-喹唑啉 -6-基 丙烯酰胺 (CXM-16);  N-{4-[l-(3-fluoro-benzyl)carbazole-5-ylamino]-7-cyano-quinazoline-6-yl acrylamide (CXM-16);
N-{7- [(四氢 -2H-吡喃 -4-基) -甲硫基] -4-[1-(3-氟-苄基) 吲唑 -5-基胺基] - 喹唑啉 -6-基 丙烯酰胺 (CXM-17);  N-{7-[(tetrahydro-2H-pyran-4-yl)-methylthio]-4-[1-(3-fluoro-benzyl)oxazol-5-ylamino]-quinazoline Porphyrin-6-yl acrylamide (CXM-17);
N-{4-[l-(3-氟-苄基） 吲唑 -5-基胺基] -喹唑啉 -7-基 丙烯酰胺 N-{4-[l-(3-Fluoro-benzyl)carbazole-5-ylamino]-quinazoline-7-yl acrylamide
(CXM-18); (CXM-18);
( - {4-[1-(3-氟-苄基) 吲唑 -5-基胺基 ]-6-甲氧基-喹唑啉 -7-基}-4-二 甲基胺基-丁 -2-烯酰胺 (CXM-19);  (-{4-[1-(3-Fluoro-benzyl)oxazol-5-ylamino]-6-methoxy-quinazolin-7-yl}-4-dimethylamino-butyl -2-enamide (CXM-19);
( - {4-[1-(3-氟-苄基) 吲唑 -5-基胺基 ]-6-乙氧基-喹唑啉 -7-基}-4-二 甲基胺基-丁 -2-烯酰胺 (CXM-20);  (-{4-[1-(3-Fluoro-benzyl)oxazol-5-ylamino]-6-ethoxy-quinazolin-7-yl}-4-dimethylamino-butyl -2-enamide (CXM-20);
( - {4-[1-(3-氟-苄基) 吲唑 -5-基胺基 ]-6-(2-甲氧基乙氧基) -喹唑啉 -7- 基}-4-二甲基胺基-丁 -2-烯酰胺 (CXM-21);  (-{4-[1-(3-Fluoro-benzyl)oxazol-5-ylamino]-6-(2-methoxyethoxy)-quinazolin-7-yl}-4- Dimethylamino-but-2-enamide (CXM-21);
N-{4-[l-(3-氟-苄基) 吲唑 -5-基胺基 ]-6-(2-甲氧基乙氧基) -喹唑啉 -7- 基 丙烯酰胺 （CXM-22);  N-{4-[1-(3-Fluoro-benzyl)oxazol-5-ylamino]-6-(2-methoxyethoxy)-quinazolin-7-yl acrylamide (CXM -twenty two);
W-{4-[l-(3-氟-苄基) 吲唑 -5-基胺基 ]-6-[3-(4-吗啉代)丙氧基] -喹唑啉 -7- 基 丙烯酰胺 （CXM-23);  W-{4-[l-(3-Fluoro-benzyl)oxazol-5-ylamino]-6-[3-(4-morpholino)propoxy]-quinazolin-7-yl Acrylamide (CXM-23);
( )-W-{4-[l-(3-氟-苄基) 吲唑 -5-基胺基 ]-6-[3-(4-吗啉代)丙氧基] -喹唑 啉 -7-基}-4-二甲基胺基-丁 -2-烯酰胺 （CXM-24) ( ) -W-{4-[l-(3-Fluoro-benzyl)oxazol-5-ylamino]-6-[3-(4-morpholino)propoxy]-quinazoline- 7 -yl}-4-dimethylamino-but-2-enamide (CXM-24)
( )-N-{6-[( -四氢呋喃 -3-基氧基 ]-4-[1-(3-氟-苄基) 吲唑 -5-基胺基] -喹 唑啉 -7-基}-4-二甲基胺基-丁 -2-烯酰胺 （CXM-25);  ( )-N-{6-[(-tetrahydrofuran-3-yloxy]-4-[1-(3-fluoro-benzyl)oxazol-5-ylamino]-quinazolin-7-yl }-4-dimethylamino-but-2-enamide (CXM-25);
( - {4-[1-(3-氟-苄基) 吲唑 -5-基胺基 ]-6-腈基-喹唑啉 -7-基}-4-二甲 基胺基-丁 -2-烯酰胺 （CXM-26); ( -{4-[1-(3-Fluoro-benzyl)oxazol-5-ylamino]-6-cyano-quinazolin-7-yl}-4-dimethylamino-butyl- 2-enamide (CXM-26);
( -N-{6- [(四氢 -2H-吡喃 -4-基) -甲硫基] -4-[1-(3-氟-苄基) 吲唑 -5-基胺 基] -喹唑啉 -7-基}-4-二甲基胺基-丁 -2-烯酰胺 （CXM-27); (-N-{6-[(tetrahydro-2H-pyran-4-yl)-methylthio]-4-[1-(3-fluoro-benzyl)oxazol-5-ylamino] - Quinazoline- 7 -yl}-4-dimethylamino-but-2-enamide (CXM-27);
( - {4-[1-(3-氟-苄基) 吲唑 -5-基胺基] -喹唑啉 -7-基}-4-二甲基胺基- 丁 -2-烯酰胺 （CXM-28); (-{4-[1-(3-Fluoro-benzyl)oxazol-5-ylamino]-quinazolin-7-yl}-4-dimethylamino- But-2-enamide (CXM-28);
N-{4-[l-(3-氟 -苄基 吲唑 -5-基胺基 ]-6-甲氧基-喹唑啉 -7-基 丙烯酰胺 (CXM-29);  N-{4-[1-(3-Fluoro-benzylcarbazole-5-ylamino]-6-methoxy-quinazolin-7-yl acrylamide (CXM-29);
N-{4-[l-(3-氟-苄基) 吲唑 -5-基胺基 ]-6-乙氧基-喹唑啉 -7-基 丙烯酰胺 (CXM-30);  N-{4-[1-(3-Fluoro-benzyl)carbazole-5-ylamino]-6-ethoxy-quinazolin-7-yl acrylamide (CXM-30);
N-{6-[( -四氢呋喃 -3-基氧基 ]-4-[1-(3-氟-苄基) 吲唑 -5-基胺基] -喹唑啉 -7-基} -丙烯酰胺 （CXM-31);  N-{6-[(-tetrahydrofuran-3-yloxy]-4-[1-(3-fluoro-benzyl)oxazol-5-ylamino]-quinazolin-7-yl}-propene Amide (CXM-31);
N-{4-[l-(3-氟 -苄基 )-1^-吲唑 -5-基胺基 ]-6-腈基-喹唑啉 -7-基 丙烯酰胺 (CXM-32);  N-{4-[1-(3-Fluoro-benzyl)-1^-oxazol-5-ylamino]-6-cyano-quinazolin-7-yl acrylamide (CXM-32);
N-{6- [(四氢 -2H-吡喃 -4-基) -甲硫基] -4-[1-(3-氟-苄基) 吲唑 -5-基胺基] - 喹唑啉 -7-基} -丙烯酰胺 （CXM-33)。  N-{6-[(tetrahydro-2H-pyran-4-yl)-methylthio]-4-[1-(3-fluoro-benzyl)oxazol-5-ylamino]-quinazoline Porphyrin-7-yl}-acrylamide (CXM-33).
本发明所述的化合物可以是互变异构体、立体异构体或其药学上可接受的 盐的形式。  The compounds of the present invention may be in the form of tautomers, stereoisomers or pharmaceutically acceptable salts thereof.
在本发明中， 除非另外具体说明， 下面的术语具有下述含义。  In the present invention, the following terms have the following meanings unless specifically stated otherwise.
"卤素"指氟、 氯、 溴和碘， 优选氟、 氯或溴。  "Halogen" means fluoro, chloro, bromo and iodo, preferably fluoro, chloro or bromo.
"CM垸基"指甲基、 乙基、 丙基、 异丙基、 丁基、 异丁基、 仲丁基和叔丁 基； 优选甲基、 乙基、 丙基、 异丙基或丁基； 更优选甲基。 "C M thiol" means methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl and tert-butyl; preferably methyl, ethyl, propyl, isopropyl or butyl More preferably a methyl group.
"CM垸氧基 "指甲氧基、 乙氧基、 正丙氧基、 异丙氧基、 正丁氧基、 异丁 氧基、 仲丁氧基和叔丁氧基； 优选甲氧基、 乙氧基、 丙氧基、 异丙氧基或丁氧 基； 更优选甲氧基。 "C M methoxy" methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy and tert-butoxy; preferably methoxy, Ethoxy, propoxy, isopropoxy or butoxy; more preferably methoxy.
"酯基"指甲酸酯基、 乙酸酯基、 丙酸酯基、 丁酸酯基等， 优选乙酸酯基。 The "ester group" nail acrylate group, acetate group, propionate group, butyrate group or the like is preferably an acetate group.
"酰胺基"指甲酰胺基、 乙酰胺基、 丙酰胺基等； 优选 α、 β-不饱和丙酰胺。"Amido" nail amide group, acetamide group, propionamide group, etc.; α, β-unsaturated propionamide is preferred.
"磺酰胺基"指甲磺酰胺基、 乙基磺酰胺基、 丙基磺酰胺基、 异丙基磺酰胺 基等； 优选甲磺酰胺基。 "Sulfolamido" is a sulfonamide group, an ethylsulfonylamino group, a propylsulfonylamino group, an isopropylsulfonylamino group or the like; preferably a methanesulfonylamino group.
"苯基 "可以是被垸基、 垸氧基、 垸氧基甲基、 酯基、 磺酸酯基、 羟基等取 代的苯基， 优选垸氧基、 垸氧基甲基、 酯基、 羟基取代的苯基。  The "phenyl group" may be a phenyl group substituted with a mercapto group, an anthracenyloxy group, a nonyloxymethyl group, an ester group, a sulfonate group, a hydroxyl group or the like, preferably a decyloxy group, a decyloxymethyl group, an ester group, or a hydroxyl group. Substituted phenyl.
"呋喃基"包括 2-呋喃基和 3-呋喃基。 "四氢呋喃基 "包括 2-四氢呋喃基和 3-四氢呋喃基。 还包括被垸基、 垸氧基、 垸氧基甲基、 酯基、 磺酸酯基、 羟基 等基团取代的呋喃基和四氢呋喃基。  "Furanyl" includes 2-furyl and 3-furyl. "Tetrahydrofuranyl" includes 2-tetrahydrofuranyl and 3-tetrahydrofuranyl. Also included are furyl and tetrahydrofuranyl groups substituted with groups such as decyl, decyloxy, decyloxymethyl, ester, sulfonate, hydroxy, and the like.
"吡喃基"包括 2-吡喃基、 3-吡喃基和 4-吡喃基。 "四氢吡喃基 "包括 2-四 氢吡喃基、 3-四氢吡喃基和 4-四氢吡喃基。 还包括被垸基、 垸氧基、 垸氧基甲 基、 酯基、 磺酸酯基、 羟基等基团取代的吡喃基和四氢吡喃基。  "Pyryl" includes 2-pyranyl, 3-pyranyl and 4-pyranyl. "Tetrahydropyranyl" includes 2-tetrahydropyranyl, 3-tetrahydropyranyl and 4-tetrahydropyranyl. Also included are pyranyl and tetrahydropyranyl groups substituted with a group such as a mercapto group, a decyloxy group, a decyloxymethyl group, an ester group, a sulfonate group, a hydroxyl group or the like.
"噻吩基"包括 2-噻吩基和 3-噻吩基。还包括被垸基、垸氧基、垸氧基甲基、 酯基、 磺酸酯基、 羟基等取代的噻吩基。  "Thiophenyl" includes 2-thienyl and 3-thienyl. Also included are thienyl groups substituted with mercapto, decyloxy, decyloxymethyl, ester, sulfonate, hydroxy, and the like.
"立体异构体 "包括顺反异构体、 对映异构体和构象异构体。 "药学上可接受的盐 "是指相对无毒的本发明式 （I ) 化合物与有机酸或无 机形成的酸加成盐。这些盐可在最后分离和提纯化合物过程中原位制备， 或者 通过使化合物以其游离碱形式与适宜的无机或有机酸反应而制成。所述无机酸 或有机酸选自氢溴酸、 盐酸、 硫酸、 亚硫酸、 乙酸、 草酸、 戊酸、 油酸、 棕榈 酸、 硬脂酸、 月桂酸、 硼酸、 苯甲酸、 乳酸、 磷酸、 甲苯甲酸、 柠檬酸、 马来 酸、 富马酸、 琥珀酸、 酒石酸、 苯甲酸、 甲磺酸、 葡萄糖酸、 乳糖酸和月桂基 磺酸。 "Stereoisomers" include cis and trans isomers, enantiomers and conformational isomers. "Pharmaceutically acceptable salt" refers to a relatively non-toxic acid addition salt of a compound of formula (I) of the present invention with an organic acid or inorganic. These salts can be prepared in situ during the final separation and purification of the compound, or by reacting the compound in its free base form with a suitable inorganic or organic acid. The inorganic or organic acid is selected from the group consisting of hydrobromic acid, hydrochloric acid, sulfuric acid, sulfurous acid, acetic acid, oxalic acid, valeric acid, oleic acid, palmitic acid, stearic acid, lauric acid, boric acid, benzoic acid, lactic acid, phosphoric acid, toluene. Formic acid, citric acid, maleic acid, fumaric acid, succinic acid, tartaric acid, benzoic acid, methanesulfonic acid, gluconic acid, lactobionic acid and lauryl sulfonic acid.
根据本发明的另一方面，提供一种含有本发明的化合物以及药学上可接受 的载体的药物组合物。  According to another aspect of the invention, there is provided a pharmaceutical composition comprising a compound of the invention and a pharmaceutically acceptable carrier.
本发明的化合物可以单独或者相互结合给药，也可以与其他药学上可接受 的活性化合物联合给药。 所述化合物可以药物组合物的形式通过口服、 直肠、 肠胃外、 局部给药施予有需要的人或动物。  The compounds of the present invention may be administered alone or in combination with each other or in combination with other pharmaceutically acceptable active compounds. The compound can be administered to a human or animal in need thereof by oral, rectal, parenteral, or topical administration in the form of a pharmaceutical composition.
所述药物组合物通常根据常规方法将活性化合物与一种或多种药学上可 接受的载体混合， 制成合适的剂型。所述药学上可接受的载体指药学领域常规 的惰性载体。  The pharmaceutical compositions are usually prepared by mixing the active compound with one or more pharmaceutically acceptable carriers according to conventional methods. The pharmaceutically acceptable carrier refers to a conventional inert carrier in the pharmaceutical art.
例如， 用于口服的固体剂型包括胶囊剂、 片剂、 丸剂、 散剂和颗粒剂等。 除了活性化合物外，药物组合物可包含以下一种或多种载体：稀释剂，如淀粉、 乳糖、 蔗糖、 葡萄糖、 甘露醇或硅酸； 粘合剂， 如羟甲基纤维素、 藻酸盐、 明 胶、 聚乙烯基吡咯垸酮、 蔗糖和阿拉伯胶； 分散剂， 如聚乙二醇、 聚乙烯吡咯 垸酮； 保湿剂， 例如甘油； 崩解剂， 如琼脂、碳酸钙、 马铃薯淀粉或木薯淀粉、 藻酸、 某些复合硅酸盐和碳酸钠； 缓溶剂， 如石蜡； 吸收加速剂， 如季胺化合 物； 润湿剂， 如鲸蜡醇和单硬脂酸甘油酯； 吸附剂， 如高岭土； 润滑剂， 例如 滑石粉、 硬脂酸钙、 硬脂酸镁、 固体聚乙二醇、 十二垸基硫酸钠； 其他辅剂， 如缓冲剂、着色剂、芳香剂、甜味剂等。此外， 可包含包衣材料制备包衣制剂。 可包含聚合物质和蜡类物质等制备缓释制剂，使活性化合物以延迟的方式在消 化道内的某一部分中释放。  For example, solid dosage forms for oral administration include capsules, tablets, pills, powders, granules and the like. In addition to the active compound, the pharmaceutical composition may comprise one or more of the following carriers: diluents such as starch, lactose, sucrose, glucose, mannitol or silicic acid; binders such as hydroxymethylcellulose, alginate , gelatin, polyvinylpyrrolidone, sucrose and gum arabic; dispersing agents such as polyethylene glycol, polyvinylpyrrolidone; humectants such as glycerin; disintegrating agents such as agar, calcium carbonate, potato starch or tapioca Starch, alginic acid, certain complex silicates and sodium carbonate; slow solvents such as paraffin; absorption accelerators, such as quaternary amine compounds; wetting agents, such as cetyl alcohol and glyceryl monostearate; adsorbents, such as kaolin Lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium dodecyl sulfate; other adjuvants such as buffers, colorants, fragrances, sweeteners, and the like. Further, a coating material may be included to prepare a coating preparation. A sustained release preparation may be prepared by including a polymeric substance and a waxy substance to release the active compound in a certain portion of the digestive tract in a delayed manner.
用于口服给药的液体剂型包括乳液、 溶液、 悬浮液、 糖桨或酊剂。 除了活 性化合物外，药物组合物可包含以下一种或多种载体：稀释剂如水或其它溶剂、 增溶剂和乳化剂， 如乙醇、 异丙醇、 碳酸乙酯、 乙酸乙酯、 丙二醇、 1,3-丁二 醇、 二甲基甲酰胺以及油， 特别是棉籽油、 花生油、 玉米胚油、 橄榄油、 蓖麻 油和芝麻油或这些物质的混合物； 助悬剂， 如乙氧基化异十八垸醇、 聚氧乙烯 山梨醇和脱水山梨醇酯、 微晶纤维素、 甲醇铝和琼脂。 此外可包含辅剂， 如润 湿剂、 乳化剂、 悬浮剂、 甜味剂、 矫味剂和香料。  Liquid dosage forms for oral administration include emulsions, solutions, suspensions, dragees or elixirs. In addition to the active compound, the pharmaceutical composition may comprise one or more of the following carriers: diluents such as water or other solvents, solubilizers and emulsifiers such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1, 3-butanediol, dimethylformamide and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances; suspending agents, such as ethoxylated iso-eighteen Sterols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar. In addition, adjuvants such as wetting agents, emulsifying agents, suspending agents, sweetening agents, flavoring agents and perfumes may be included.
用于肠胃外给药的剂型包括静脉内、 皮下、肌内注射制剂。 除了活性化合 物外， 药物组合物可包含生理上可接受的无菌含水或无水溶液、 分散液、 悬浮 液或乳液， 和用于重新溶解成无菌的可注射溶液或分散液的无菌粉末。适宜的 含水和非水载体、 稀释剂、 溶剂包括水、 乙醇、 多元醇及其适宜的混合物。 Dosage forms for parenteral administration include intravenous, subcutaneous, intramuscular injection preparations. In addition to the active compound, the pharmaceutical compositions may comprise physiologically acceptable sterile aqueous or nonaqueous solutions, dispersions, suspensions A liquid or emulsion, and a sterile powder for reconstitution into a sterile injectable solution or dispersion. Suitable aqueous and nonaqueous vehicles, diluents, and solvents include water, ethanol, polyols, and suitable mixtures thereof.
用于局部给药的剂型包括软膏剂、 散剂、 喷射剂、 滴剂和吸入剂。 活性成 分在无菌条件下与生理上可接受的载体及任何防腐剂、缓冲剂， 或必要时可能 需要的推进剂一起混合。  Dosage forms for topical administration include ointments, powders, propellants, drops, and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants which may be required if necessary.
根据本发明的另一方面，提供本发明的化合物或药物组合物在制备抗肿瘤 药物中的应用， 所述肿瘤优选为由蛋白络氨酸激酶介导的疾病， 例如乳腺癌、 非小细胞肺癌、 卵巢癌、 胃癌、 胰腺癌等。 具体实施方式  According to another aspect of the present invention, there is provided a use of a compound or a pharmaceutical composition of the present invention for the preparation of an antitumor drug, preferably a disease mediated by a protein tyrosine kinase, such as breast cancer, non-small cell lung cancer , ovarian cancer, stomach cancer, pancreatic cancer, etc. Detailed ways
通过以下实施例对本发明作进一步详细说明，但不应理解为限定本发明的 范围。  The invention is further illustrated by the following examples, which are not to be construed as limiting the scope of the invention.
实施例 1 制备 N-{4-[l-(3-氟 -苄基 )-1^-吲唑 -5-基胺基] -喹唑啉 -6-基+丙烯酰胺 (化合物 CXM-01) Example 1 Preparation of N-{4-[1-(3-fluoro-benzyl)-1^-oxazol-5-ylamino]-quinazolin-6-yl+acrylamide (Compound CXM-01)
Figure imgf000009_0001
Figure imgf000009_0001
将 5-硝基吲唑 (163.1g, lmol)、 碳酸铯 (358.4g, l. lmol)溶于 DMF(0.5L)， 于 70-80°C反应 2h， 然后加入 3-氟苄基溴 (225.6g, 1.2mol)继续反应， 直至薄层层 析检测反应完全。 将反应液冷却至室温， 过滤， 并用 DMF(0.3L)洗涤滤饼。 滤 液在 15-20°C时加入水 (0.13-0.15L), 放置 4h， 过滤， 滤饼依次用 DMF/水混合 物 (0.2L, v/v 2: 1)、 水 (0.2L)和冷的 CAN (硝酸铈铵 )/水混合物 (0.15L, v/v 3: 1)洗 涤， 在低于 45。C的温度下干燥， 得 5-硝基 -1-(3-氟苄基) - Π引唑结晶 (化合物 1)， 收率 56% 1H-NMR(400 MHz, CDC13) δ 5.62 (s, 2Η)， 6.85 (d, 1Η, J = 9.4 Hz), 6.93 (m, 2H), 7.28 (m, 1H), 7.40 (d, 1H, / = 9.2 Hz), 8.21 (dd, 1H, / = 10 Hz and 2 Hz), 8.24 (s, 1H), 8.70 (d, lH, / = 2 Hz); MS: 272(M+H)+; HPLC保留时间： 6.95 min (YMC ODS-A 3μηι, 4.6x50mm柱， lOmin梯度， 2.5 ml/min)。 5-Nitrocarbazole (163.1 g, 1 mol), cesium carbonate (358.4 g, 1.1 mol) was dissolved in DMF (0.5 L), reacted at 70-80 ° C for 2 h, then 3-fluorobenzyl bromide ( The reaction was continued at 225.6 g, 1.2 mol) until the reaction was complete by thin layer chromatography. The reaction solution was cooled to room temperature, filtered, and the filter cake was washed with DMF (0.3L). The filtrate was added with water (0.13-0.15 L) at 15-20 ° C, placed for 4 h, filtered, and the filter cake was sequentially treated with DMF/water mixture (0.2 L, v/v 2: 1), water (0.2 L) and cold. CAN (ammonium nitrate) / water mixture (0.15L, v/v 3: 1) wash Polyester, at less than 45. Drying at a temperature of C gives 5-nitro-1-(3-fluorobenzyl)- oxazole crystals (Compound 1) in a yield of 56% 1 H-NMR (400 MHz, CDC1 3 ) δ 5.62 (s , 2Η), 6.85 (d, 1Η, J = 9.4 Hz), 6.93 (m, 2H), 7.28 (m, 1H), 7.40 (d, 1H, / = 9.2 Hz), 8.21 (dd, 1H, / = 10 Hz and 2 Hz), 8.24 (s, 1H), 8.70 (d, lH, / = 2 Hz); MS: 272 (M+H) + ; HPLC retention time: 6.95 min (YMC ODS-A 3μηι, 4.6 X50 mm column, lOmin gradient, 2.5 ml/min).
在氢化反应釜中加入化合物 l(271.2g, 111101)，^^(0.81^，5%钯碳（2.7 化 合物 1的 0.1wt% ) 作为催化剂， 于 30-35°C、 15psi反应 2h。 反应液冷却至室 温， 过滤， 用 THF(O. IL)洗涤滤饼。 将滤饼加入 THF(0.5L)中搅拌， 过滤。 合 并滤液， 常压蒸馏至所需体积 (0.55L), 并在 55-60°C 下在 lh 以内加入庚垸 (1.5L), 得到桨状物。 将桨状物在 1.5h内冷却至室温， 放置 lh， 过滤， 滤饼用 THF/庚垸 (1L， v/v 1 : 4)洗涤， 40°C鼓风干燥， 得 1-(3-氟-苄基) 吲唑 -5-基 胺 (化合物 2)， 收率 80%。 mp: 130°C。 HPLC保留时间： 9.05min。  In the hydrogenation reactor, compound l (271.2 g, 111101), ^(0.81, 5% palladium on carbon (2.7 wt% of compound 1) was used as a catalyst, and reacted at 30-35 ° C, 15 psi for 2 h. The mixture was cooled to room temperature, filtered, and the filter cake was washed with THF (EtOAc). The filter cake was stirred in THF (0.5 L) and filtered. The filtrate was combined and distilled to the desired volume (0.55 L) at 55- Ginger (1.5 L) was added at a temperature of 60 ° C to obtain a paddle. The paddle was cooled to room temperature in 1.5 h, allowed to stand for 1 h, filtered, and the filter cake was THF / heptane (1 L, v/v 1 : 4) Washing, drying at 40 ° C to obtain 1-(3-fluoro-benzyl)oxazol-5-ylamine (Compound 2), yield 80%. mp: 130 ° C. HPLC retention time : 9.05min.
将 6-硝基喹唑啉酮 (191. lg, lmol)和三氯氧磷 (0.15L)于 105-110。C回流反应 3h， 反应液缓慢倒入冰水 (0.9L)中析出鳞片状固体， 过滤， 干燥， 得 4-氯 -6-硝 基喹唑啉 (化合物 3)，收率 80%。 ^-NMR OO MHz, CDC13): δ 9.20 (s, 2Η), 8.72 (dd, 1H, J = 2.57Hz, 9.16 Hz), 8.25 (d, 1H, J = 9.16 Hz). 6-Nitroquinazolinone (191. lg, 1 mol) and phosphorus oxychloride (0.15 L) were at 105-110. C was refluxed for 3 h, and the reaction mixture was poured into ice water (0.9 L) to precipitate a flaky solid, which was filtered and dried to give 4-chloro-6-nitroquinazoline (Compound 3) in a yield of 80%. ^-NMR OO MHz, CDC1 3 ): δ 9.20 (s, 2Η), 8.72 (dd, 1H, J = 2.57 Hz, 9.16 Hz), 8.25 (d, 1H, J = 9.16 Hz).
将化合物 2(241.2g, lmol)和化合物 3(209.6g, lmol)溶于异丙醇 (1.5L),回流 反应 3h， 析出黄色固体， 过滤， 真空干燥， 得黄色固体 N-[l-(3-氟 -苄基 )-1^- 吲唑 -5-基] -6-硝基喹唑啉 -4-胺 (；化合物 4)， 收率 73%。 ^-NMRWOO MHz, CDC13): δ 10.08 (s, br, 1H), 8.76-8.70 (m, 2H), 8.36 (s, 1H), 8.27-8.21 (m, 1H), 8.14-8.05 (m, 1H), 7.35-7.32 (m, 1H), 7.12-7.03 (m, 1H), 7.05-6.96 (m, 1H), 6.83-6.80 (m, 1H), 6.78-6.68 (m, 2H),6.54-6.51 (m, 1H), 4.99 (s, 2H). The compound 2 (241.2 g, 1 mol) and the compound 3 (209.6 g, 1 mol) were dissolved in isopropyl alcohol (1.5 L), and refluxed for 3 h to precipitate a yellow solid which was filtered and dried in vacuo to give a yellow solid N-[l-( 3-Fluoro-benzyl)-1^-carbazol-5-yl]-6-nitroquinazolin-4-amine (Compound 4), yield 73%. ^-NMRWOO MHz, CDC1 3 ): δ 10.08 (s, br, 1H), 8.76-8.70 (m, 2H), 8.36 (s, 1H), 8.27-8.21 (m, 1H), 8.14-8.05 (m, 1H), 7.35-7.32 (m, 1H), 7.12-7.03 (m, 1H), 7.05-6.96 (m, 1H), 6.83-6.80 (m, 1H), 6.78-6.68 (m, 2H), 6.54 6.51 (m, 1H), 4.99 (s, 2H).
将化合物 4(414.4g, lmol)，铁粉 (279.2g, 5mol)加入冰醋酸 (0.05L)和甲醇 (1L) 中， 回流反应 4h， 反应液过滤除去铁粉， 滤液加入乙酸乙酯 (0.5L), 然后依次 用适量碳酸氢钠溶液和水洗涤， 分离有机相， 无水硫酸镁干燥， 浓缩， 得黄色 固体 ^-[1-(3-氟-苄基) 吲唑 -5-基] -喹唑啉 -4,6-二胺 (化合物 5)， 收率 64%。 1H-NMR(400 MHz, DMSO): δ 9.43 (s, 1Η), 8.32 (s, 1H), 8.14-8.10 (m, 1H), 7.76-7.71 (m, 1H), 7.35-7.32 (m, 1H), 7.13-7.10 (m, 1H), 7.08-6.99 (m, 3H), 6.83-6.70 (m, 3H), 6.54-6.44 (m, 1H), 5.19 (s, 2H). Compound 4 (414.4 g, 1 mol), iron powder (279.2 g, 5 mol) was added to glacial acetic acid (0.05 L) and methanol (1 L), and refluxed for 4 h. The reaction mixture was filtered to remove iron powder. L), and then washed with an appropriate amount of sodium bicarbonate solution and water, and the organic phase is separated, dried over anhydrous magnesium sulfate, and evaporated to give a yellow solid (-[1-(3-fluoro-benzyl) oxazol-5-yl] - quinazoline-4,6-diamine (compound 5), yield 64%. 1 H-NMR (400 MHz, DMSO): δ 9.43 (s, 1 Η), 8.32 (s, 1H), 8.14-8.10 (m, 1H), 7.76-7.71 (m, 1H), 7.35-7.32 (m, (H, 3H)
在 0-5。C温度下向化合物 5(384.4g, lmol)中加入三乙胺 (151.8g, 1.5mol)， 丙烯酰氯 (90.5g, lmol), 四氢呋喃 (1.3L), 缓慢升至室温， 反应 3h， 反应液用 乙酸乙酯 (1.5L)萃取， 分离有机相， 无水硫酸镁干燥， 浓缩， 浓缩物经硅胶柱 层析分离 (流动相： 乙酸乙酯与石油醚：)，得淡黄色固体 N-{4-[l-(3-氟 -苄基 吲唑 -5-基胺基]-喹唑啉 -6-基- } -丙烯酰胺 (标题化合物 CXM-01), 收率 70%。 1H-NMR(400 MHz, CDC13): δ 8.38 (s, 1Η), 8.22-8.14 (m, 2H), 8.14-8.10 (m, 1H), 7.99-7.94 (m, 1H), 7.35-7.32 (m, 1H), 7.12-7.05 (m, 2H), 6.83-6.77 (m, 3H), 6.54-6.52 (m, 1H), 6.48-6.36 (d, J =5.9 Hz, 2H), 5.71-5.67 (t, J =5.9 Hz, 2H), 6.5.09 (s, 2H). At 0-5. To the compound 5 (384.4 g, 1 mol), triethylamine (151.8 g, 1.5 mol), acryloyl chloride (90.5 g, 1 mol), tetrahydrofuran (1.3 L), slowly warmed to room temperature for 3 h, reaction solution Extracted with ethyl acetate (1.5 L), EtOAc (EtOAc mjjjHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHH 4- [l- (3- fluoro - benzyl-indazol-5-yl amino] - quinazolin-6-yl -} - acrylamide (title compound CXM-01), yield 70% 1 H- NMR (400 MHz, CDC1 3 ): δ 8.38 (s, 1 Η), 8.22-8.14 (m, 2H), 8.14-8.10 (m, 1H), 7.99-7.94 (m, 1H), 7.35-7.32 (m, 1H), 7.12-7.05 (m, 2H), 6.83-6.77 (m, 3H), 6.54-6.52 (m, 1H), 6.48-6.36 (d , J = 5.9 Hz, 2H), 5.71-5.67 (t, J = 5.9 Hz, 2H), 6.5.09 (s, 2H).
实施例 2 制备^) -N-{4-[l-(3-氟-苄基：) 吲唑 -5-基胺基 ]-7-甲氧基-喹唑啉 -6- 基- }-4 Example 2 Preparation of ^)-N-{4-[1-(3-Fluoro-benzyl:)oxazol-5-ylamino]-7-methoxy-quinazolin-6-yl- }- 4
Figure imgf000011_0001
Figure imgf000011_0001
将 2-氨基 -4-氯-苯甲酸 (171.6g, lmol)溶于甲酰胺 (972.8g, 21.6mol)， 回流反 应 5h， 反应液过滤， 干燥滤饼， 得 7-氯喹唑酮 (化合物 6)。  2-Amino-4-chloro-benzoic acid (171.6 g, 1 mol) was dissolved in formamide (972.8 g, 21.6 mol), refluxed for 5 h, the reaction mixture was filtered, and the filter cake was dried to give 7-chloroquinazolidone (Compound 6) ).
将化合物 6(180.6g, lmol)在冰浴下缓慢加入混酸 (0.4L,含有浓硫酸和发烟 硝酸， v/v 1 : 1)中，加热至 90°C反应 4h。将澄清的反应液缓慢倒入冰水 (3L)中， 析出浅黄色沉淀， 过滤， 滤饼用热冰醋酸 (2L)重结晶， 得 6-硝基 -7-氯喹唑酮晶 体 (化合物 7)。  Compound 6 (180.6 g, 1 mol) was slowly added to the mixed acid (0.4 L, containing concentrated sulfuric acid and fuming nitric acid, v/v 1 : 1) in an ice bath, and heated to 90 ° C for 4 h. The clarified reaction solution was slowly poured into ice water (3 L) to precipitate a pale yellow precipitate, which was filtered, and the filter cake was recrystallized from hot glacial acetic acid (2 L) to give crystals of 6-nitro-7-chloroquinazolidone (Compound 7) .
将化合物 7(225.6g, lmol)与三氯氧磷 (0.375L)回流反应 4h， 反应液倒入冰 水中， 过滤， 干燥滤饼， 得 6-硝基 -4,7-二氯喹唑啉 (化合物 8)。  Compound 7 (225.6 g, 1 mol) and phosphorus oxychloride (0.375 L) were refluxed for 4 h, and the reaction solution was poured into ice water, filtered, and the filter cake was dried to give 6-nitro-4,7-dichloroquinazoline ( Compound 8).
将化合物 8(170.1g, 0.7mol)溶于异丙醇 (0.75L)中， 加入 1-(3-氟-苄基) 口引挫 -5-基胺 (化合物 2， 168.8g， 0.7mol) , 回流反应 4h， 过滤， 真空干燥滤饼， 得 W-[l-(3-氟-苄基) 吲唑 -5-基] -7-氯 -6-硝基-喹唑啉 -4-胺 (化合物 9)。  Compound 8 (170.1 g, 0.7 mol) was dissolved in isopropanol (0.75 L), and 1-(3-fluoro-benzyl) was added to finch-5-ylamine (Compound 2, 168.8 g, 0.7 mol) , refluxing for 4 h, filtration, vacuum drying of the filter cake to give W-[l-(3-fluoro-benzyl)carbazol-5-yl]-7-chloro-6-nitro-quinazolin-4-amine (Compound 9).
将化合物 9(448.8g， lmol)溶于无水 THF(2.5L)中， 加入甲醇钠 (63.74g， 1.18mol) , 回流反应 18h。 反应液加入冰醋酸 (70.86g， 1.18mol) , 水洗， 然后用 乙酸乙酯 (0.5L)萃取 3次， 有机相减压浓缩， 干燥， 得 N-[l -(3-氟-苄基) 吲 唑 -5-基 ]-7-甲氧基 -6-硝基-喹唑啉 -4-胺 (；化合物 10)。 Compound 9 (448.8 g, 1 mol) was dissolved in dry THF (2.5 L) and sodium methoxide (63.74 g, 1.18 mol), reflux reaction for 18 h. The reaction solution was added with glacial acetic acid (70.86 g, 1.18 mol), washed with water, and then extracted three times with ethyl acetate (0.5 L). The organic phase was concentrated under reduced pressure and dried to give N-[l-(3-fluoro-benzyl). Oxazol-5-yl]- 7 -methoxy-6-nitro-quinazolin-4-amine (Compound 10).
将化合物 10(444.4g, lmol)、铁粉 (279.2g, 5mol)、冰醋酸 (0.25L)、甲醇 (2.5L) 混合， 回流反应 3h， 反应液过滤除去铁粉， 滤液用乙酸乙酯 (0.5L)稀释， 依次 用饱和碳酸氢钠溶液 (1L)和水 (0.2L)洗涤， 有机相用无水硫酸镁干燥， 浓缩， 得浅黄色固体化合物 11， 收率 73%。  Compound 10 (444.4 g, 1 mol), iron powder (279.2 g, 5 mol), glacial acetic acid (0.25 L), and methanol (2.5 L) were mixed, and refluxed for 3 hours. The reaction liquid was filtered to remove iron powder, and the filtrate was evaporated. The mixture was diluted with EtOAc (3 mL).
将化合物 ll(414.4g, lmol)溶于无水 THF(2.5L)中，于 0°C依次滴加二异丙 基乙基胺 (0.15L)和 3-N,N-二甲氨基甲基丙烯酰氯 (147.6g, lmol) ,于 0-5°C反应 3h。 向反应液加入乙酸乙酯-水 (v/v， 4: 1)的混合物 (1L)， 水层用乙酸乙酯 (0.5L) 洗涤 3 次， 合并有机相， 无水硫酸镁干燥， 浓缩， 硅胶柱层析分离， 得固体 ( - {4-[1-(3-氟-苄基) 吲唑 -5-基胺基 ]-7-甲氧基-喹唑啉 -6-基 -}-4- (二甲基 氨基）丁 -2-烯酰胺 (；标题化合物 CXM-02) , 收率 67%。 ^-NMR^OO MHz , CDC13) : δ 8.36 (s, 1Η), 8.20-8.14 (m, 1H), 8.12-8.10 (m, 1H), 7.50-7.46 (m, 1H), 7.36-7.33 (m, 1H), 7.10-7.05 (m, 2H), 6.83-6.77 (m, 3H), 6.75-6.73 (d, J =2.5 Hz, 1H), 6.54-6.52 (d, / =2.5 Hz, 1H), 6.38-6.36 (m, 1H), 4.99-4.94 (m, 2H), 3.83 (s, 3H), 3.03 (m, 2H), 2.27-2.18 (m, 6H). Compound ll (414.4 g, 1 mol) was dissolved in anhydrous THF (2.5 L), and diisopropylethylamine (0.15 L) and 3-N,N-dimethylaminomethyl were added dropwise at 0 °C. Acryloyl chloride (147.6 g, 1 mol) was reacted at 0-5 ° C for 3 h. A mixture of ethyl acetate-water (v/v, 4:1) (1 L) was evaporated. Silica gel column chromatography gave solid (-{4-[1-(3-fluoro-benzyl)oxazol-5-ylamino]-7-methoxy-quinazolin-6-yl-}- 4-(Dimethylamino)but-2-enamide (title compound CXM-02), yield 67%. ^-NMR^OO MHz , CDC1 3 ) : δ 8.36 (s, 1 Η), 8.20-8.14 (m, 1H), 8.12-8.10 (m, 1H), 7.50-7.46 (m, 1H), 7.36-7.33 (m, 1H), 7.10-7.05 (m, 2H), 6.83-6.77 (m, 3H) , 6.75-6.73 (d, J = 2.5 Hz, 1H), 6.54-6.52 (d, / =2.5 Hz, 1H), 6.38-6.36 (m, 1H), 4.99-4.94 (m, 2H), 3.83 (s , 3H), 3.03 (m, 2H), 2.27-2.18 (m, 6H).
实施例 3 制备^) -N- {4-[l-(3-氟-苄基:) 吲唑 -5-基胺基 ]-7-乙氧基-喹唑啉 -6- 基}—4- (二甲基胺基) - -2-烯酰胺 (化合物 CXM-03) Example 3 Preparation of ^) -N- {4-[l-(3-fluoro-benzyl:)oxazol-5-ylamino]-7-ethoxy-quinazolin-6-yl}-4 - (Dimethylamino)-2-enamide (Compound CXM-03)
Figure imgf000012_0001
Figure imgf000012_0001
制备方法类似实施例 2， 不同的是将甲醇钠替换为乙醇钠。 终产物为标题 化合物 CXM-03。 1H-NMR(400 MHz, CDC13) : δ 8.37 (s, 1H), 8.21-8.13 (m, 1H), 8.11-8.08 (m, 1H), 7.51-7.48 (m, 1H), 7.37-7.32 (m, 1H), 7.11-7.06 (m, 2H), 6.82-6.75 (m, 3H), 6.70-6.68 (d, J =2.5 Hz, 1H), 6.50-6.48 (d, J =2.5 Hz, 1H), 6.40-6.35 (m, 1H), 5.03-4.96 (m, 2H), 3.85 (s, 3H), 3.03 (m, 2H), 2.27-2.18 (m, 6H), 1.33-1.30 (m, 3H). The preparation method was similar to that of Example 2, except that sodium methoxide was replaced with sodium ethoxide. The final product was the title compound CXM-03. 1 H-NMR (400 MHz, CDC1 3 ) : δ 8.37 (s, 1H), 8.21-8.13 (m, 1H), 8.11-8.08 (m, 1H), 7.51-7.48 (m, 1H), 7.37-7.32 (m, 1H), 7.11-7.06 (m, 2H), 6.82-6.75 (m, 3H), 6.70-6.68 (d, J = 2.5 Hz, 1H), 6.50-6.48 (d, J = 2.5 Hz, 1H ), 6.40-6.35 (m, 1H), 5.03-4.96 (m, 2H), 3.85 (s, 3H), 3.03 (m, 2H), 2.27-2.18 (m, 6H), 1.33-1.30 (m, 3H) ).
实施例 4 制备 7-(2-甲氧基乙氧基) -N-[l-(3-氟-苄基) 吲唑 -5-基] -6-硝基-喹 唑啉 -4-胺 （化合物 CXM-04 ) Example 4 Preparation of 7-(2-methoxyethoxy)-N-[l-(3-fluoro-benzyl)oxazol-5-yl]-6-nitro-quinazolin-4-amine (Compound CXM-04)
Figure imgf000013_0001
Figure imgf000013_0001
4-氯 -6-硝基-喹唑啉 -7-基-乙酸酯 (267g, lmol)和 1-(3-氟-苄基) 吲唑 -5-胺 (241. lg, lmol)溶于异丙醇 (1L)中， 回流反应 3h。 反应液过滤， 滤饼用乙酸乙酯 (0.3L)洗涤， 45°C真空干燥过夜， 得浅黄色固体 4-[1-(3-氟-苄基) 吲唑 -5- 基胺基] - 6-硝基-喹唑啉 -7-基-乙酸酯 (化合物 12)，收率
Figure imgf000013_0002
4-Chloro-6-nitro-quinazolin-7-yl-acetate (267 g, 1 mol) and 1-(3-fluoro-benzyl)oxazol-5-amine (241. lg, 1 mol) The reaction was refluxed for 3 h in isopropanol (1 L). The reaction mixture was filtered, and EtOAc EtOAc (EtOAc) 6-nitro-quinazolin-7-yl-acetate (compound 12), yield
Figure imgf000013_0002
CDC13): δ 8.74 (s, 1H), 8.36-8.32 (m, 1H), 8.14-8.08 (m, 2H), 7.35-7.30 (m, 1H), 7.12-7.08 (m, 2H), 6.83-6.77 (m, 3H), 6.54-6.49 (m, 1H), 5.03-4.99 (m, 2H), 2.08 (m, 3H). CDC1 3 ): δ 8.74 (s, 1H), 8.36-8.32 (m, 1H), 8.14-8.08 (m, 2H), 7.35-7.30 (m, 1H), 7.12-7.08 (m, 2H), 6.83- 6.77 (m, 3H), 6.54-6.49 (m, 1H), 5.03-4.99 (m, 2H), 2.08 (m, 3H).
将化合物 12(472.1g, lmol)溶于甲醇 (5L)中， 滴加浓氨水 (0.5L), 室温反应 5h， 反应液过滤， 滤液浓缩， 浓缩物与滤饼合并， 用甲醇重结晶， 得黄色固体 4-[1-(3-氟-苄基：) 吲唑 -5-基胺基 ]-6-硝基-喹唑啉 -7-羟基 (化合物 13)， 收率 95% 1H-NMR(400MHz, CDC13): δ 8.60 (s, 1H), 8.36-8.32 (m, 1H), 8.14-8.10 (m, 1H), 7.76-7.72 (m, 1H), 7.35-7.32 (m, 1H), 7.12-7.05 (m, 2H), 6.83-6.77 (m, 3H), 6.54-6.50 (m, 1H), 5.05-5.00 (m, 2H). The compound 12 (472.1 g, 1 mol) was dissolved in methanol (5 L), concentrated aqueous ammonia (0.5 L) was added dropwise, and the mixture was reacted at room temperature for 5 h, the reaction liquid was filtered, and the filtrate was concentrated. The concentrate was combined with the filter cake and recrystallized from methanol. Yellow solid 4-[1-(3-fluoro-benzyl:)oxazol-5-ylamino]-6-nitro-quinazolin-7-hydroxy (compound 13), yield 95% 1 H- NMR (400MHz, CDC1 3 ): δ 8.60 (s, 1H), 8.36-8.32 (m, 1H), 8.14-8.10 (m, 1H), 7.76-7.72 (m, 1H), 7.35-7.32 (m, 1H ), 7.12-7.05 (m, 2H), 6.83-6.77 (m, 3H), 6.54-6.50 (m, 1H), 5.05-5.00 (m, 2H).
将化合物 13(430.1g, lmol)和 2-甲氧基溴乙垸 (185.9, 1.5mol)溶于 DMF(15L) 中， 加入四丁基碘化铵 (36.9g, 0.1mol)和碳酸钾 (138.2g, lmol), 60°C反应过夜， 趁热过滤，滤液减压蒸去溶剂，剩余物用甲醇重结晶，得标题化合物 CXM-04, 收率 71% 1H-NMR(400 MHz, CDC13): δ 8.66 (s, 1Η), 8.36-8.33 (m, 1H), 8.14-8. ll(m, 1H), 7.78-7.75 (m, 2H), 7.37-7.33 (m, 1H), 7.11-7.07 (m, 2H), 7.06-7.01 (m, 1H), 6.85-6.75 (m, 3H), 6.56-6.51 (m, 1H), 5.07-5.02 (m, 2H), 4.11-4.07 (m, 2H), 3.79-3.73 (m, 2H), 3.24 (s, 3H). Compound 13 (430.1 g, 1 mol) and 2-methoxybromide (185.9, 1.5 mol) were dissolved in DMF (15 L), and tetrabutylammonium iodide (36.9 g, 0.1 mol) and potassium carbonate ( The title compound CXM-04, Yield 71% 1 H-NMR (400 MHz, CDC1), mp. 3 ): δ 8.66 (s, 1Η), 8.36-8.33 (m, 1H), 8.14-8. ll(m, 1H), 7.78-7.75 (m, 2H), 7.37-7.33 (m, 1H), 7.11 -7.07 (m, 2H), 7.06-7.01 (m, 1H), 6.85-6.75 (m, 3H), 6.56-6.51 (m, 1H), 5.07-5.02 (m, 2H), 4.11-4.07 (m, 2H), 3.79-3.73 (m, 2H), 3.24 (s, 3H).
实施例 5 制备^) -Ν-{4-[1-(3-氟-苄基) 吲唑 -5-基胺基 ]-7-(3-甲氧基乙氧基) - 喹唑啉 -6-基}-4- (二甲基胺基) -丁 -2-烯酰胺 (化合物 CXM-05) Example 5 Preparation of )-{-{4-[1-(3-Fluoro-benzyl)oxazol-5-ylamino]-7-(3-methoxyethoxy)-quinazoline- 6 -yl}-4-(dimethylamino)-but-2-enamide (compound CXM-05)
Figure imgf000014_0001
Figure imgf000014_0001
制备方法类似实施例 2， 不同的是将化合物 10替换为化合物 CXM-04。 终产物为标题化合物 CXM-05。 1H-NMR(400 MHz, CDC13) : δ 8.37 (s, IH), 8.15-8.12 (m, IH), 7.50-7.45 (m, IH), 7.36-7.32 (m, IH), 7.12-7.05 (m, 2H), 6.83-6.77 (m, 3H), 6.75-6.72 (d, J =2.5 Hz, IH), 6.54-6.51 (m, IH), 6.38-6.34 (d, J =2.5 Hz, IH), 4.99-4.96 (m, 2H), 4.10-4.06 (m, 2H), 3.78-3.74 (m, 2H), 3.24 (s, 3H), 3.03-3.00 (d, / =3.8 Hz, 2H), 2.28 (s, 6H). The preparation method was similar to that of Example 2, except that Compound 10 was replaced with Compound CXM-04. The final product was the title compound CXM-05. 1 H-NMR (400 MHz, CDC1 3 ) : δ 8.37 (s, IH), 8.15-8.12 (m, IH), 7.50-7.45 (m, IH), 7.36-7.32 (m, IH), 7.12-7.05 (m, 2H), 6.83-6.77 (m, 3H), 6.75-6.72 (d, J = 2.5 Hz, IH), 6.54-6.51 (m, IH), 6.38-6.34 (d, J = 2.5 Hz, IH ), 4.99-4.96 (m, 2H), 4.10-4.06 (m, 2H), 3.78-3.74 (m, 2H), 3.24 (s, 3H), 3.03-3.00 (d, / =3.8 Hz, 2H), 2.28 (s, 6H).
实施例 6 制备 N-{4-[ l-(3-氟-苄基) -IH-吲唑 -5-基胺基 ]-7-(2-甲氧乙氧基) -喹唑 啉 -6-基} -丙烯酰胺 (化合 CXM-06) Example 6 Preparation of N-{4-[1-(3-fluoro-benzyl)-IH-indazol-5-ylamino]-7-(2-methoxyethoxy)-quinazoline-6 -yl}-acrylamide (combination CXM-06)
Figure imgf000014_0002
Figure imgf000014_0002
制备方法类似实施例 5， 不同的是将 3-N,N-二甲氨基甲基丙烯酰氯替换为 丙烯酰氯。终产物为标题化合物 CXM-06。 1H-NMR(400 MHz, CDC13) : δ 8.36 (s, IH), 8.14-8.11 (m, 2H), 7.51-7.46 (m, IH), 7.35-7.32 (m, IH), 7.12-7.05 (m, 2H), 6.84-6.78 (m, 3H), 6.54-6.51 (m, IH), 6.48-6.44 (q, / =10.1 , 16.8 Hz, IH), 6.17-6.14 (dd, / =2.1, 7.1 Hz, 2H), 5.71-5.68 (dd, / =2.1 , 7.1 Hz, 2H), 4.99-4.96 (m 2H), 4.11-4.06 (m, 2H), 3.79-3.74 (m, 2H), 3.24 (s, 3H). The preparation method was similar to that of Example 5, except that 3-N,N-dimethylaminomethacryloyl chloride was replaced with acryloyl chloride. The final product was the title compound CXM-06. 1 H-NMR (400 MHz, CDC1 3 ) : δ 8.36 (s, IH), 8.14-8.11 (m, 2H), 7.51-7.46 (m, IH), 7.35-7.32 (m, IH), 7.12-7.05 (m, 2H), 6.84-6.78 (m, 3H), 6.54-6.51 (m, IH), 6.48-6.44 (q, /=10.1, 16.8 Hz, IH), 6.17-6.14 (dd, / =2.1, 7.1 Hz, 2H), 5.71-5.68 (dd, / =2.1, 7.1 Hz, 2H), 4.99-4.96 (m 2H), 4.11-4.06 (m, 2H), 3.79-3.74 (m, 2H), 3.24 ( s, 3H).
实施例 7 制备 N-{4-[ l-(3-氟-苄基) -IH-吲唑 -5-基胺基 ]-7-[3-(4-吗啉代)丙氧基] - 喹唑啉 -6-基} -丙烯酰胺 (化合物 CXM-07) Example 7 Preparation of N-{4-[1-(3-fluoro-benzyl)-IH-indazol-5-ylamino]-7-[3-(4-morpholino)propoxy]- Quinazoline-6-yl}-acrylamide (compound CXM-07)
Figure imgf000015_0001
Figure imgf000015_0001
制备方法类似实施例 6， 不同的是将 2-甲氧基溴乙垸替换为 3-(N-吗啉) -1- 氯丙垸。终产物为标题化合物 CXM-07。 1H-NMR(400 MHz, CDC13): δ 8.34 (s, IH), 8.15-8.12 (m, 2H), 7.50-7.46 (m, IH), 7.36-7.32 (m, IH), 7.10-7.05 (m, 2H), 6.83-6.77 (m, 3H), 6.53-6.50 (m, IH), 6.46-6.42 (q, J =10.1, 16.8 Hz, IH), 6.15-6.12 (dd, / =2.1, 7.1 Hz, 2H), 5.70-5.66 (dd, / =2.1 , 7.1 Hz, 2H), 5.00-4.98 (m 2H), 3.94-3.90 (m, 2H), 3.67-3.64 (m, 4H), 2.37-2.35 (m, 6H), 1.81-1.75 (m, 2H). 实施例 8 制备 ( -N- {4-[ l-(3-氟-苄基) -IH-吲唑 -5-基胺基 ]-7-[3-(4-吗啉代)丙氧 基] -喹唑啉 -6-基}-4-二甲基胺基) -丁 -2-烯酰胺 (化合物 CXM-08) The preparation method was similar to that of Example 6, except that 2-methoxybromoacetam was replaced by 3-(N-morpholine)-1-chloroproponium. The final product was the title compound CXM-07. 1 H-NMR (400 MHz, CDC1 3 ): δ 8.34 (s, IH), 8.15-8.12 (m, 2H), 7.50-7.46 (m, IH), 7.36-7.32 (m, IH), 7.10-7.05 (m, 2H), 6.83-6.77 (m, 3H), 6.53-6.50 (m, IH), 6.46-6.42 (q, J = 10.1, 16.8 Hz, IH), 6.15-6.12 (dd, / =2.1, 7.1 Hz, 2H), 5.70-5.66 (dd, / =2.1, 7.1 Hz, 2H), 5.00-4.98 (m 2H), 3.94-3.90 (m, 2H), 3.67-3.64 (m, 4H), 2.37- 2.35 (m, 6H), 1.81-1.75 (m, 2H). Example 8 Preparation of (-N-{4-[ l-(3-fluoro-benzyl)-IH-indazol-5-ylamino] -7-[3-(4-morpholino)propoxy]-quinazoline- 6 -yl}-4-dimethylamino)-but-2-enamide (Compound CXM-08)
Figure imgf000015_0002
Figure imgf000015_0002
制备方法类似实施例 7， 不同的是将丙烯酰氯替换为 3-N,N-二甲氨基甲基 丙烯酰氯。终产物为标题化合物 CXM-08。 1H-NMR(400 MHz, CDC13) : δ 8.36 (s, IH), 8.14-8.10 (m, 2H), 7.51-7.48 (m, IH), 7.35-7.31 (m, IH), 7.12-7.08 (m, 2H), 6.84-6.78 (m, 3H), 6.54-6.50 (m, IH), 6.40-6.37 (d, J =2.5 Hz, IH), 6.29-6.26 (d, J =2.5 Hz, IH), 4.99-4.96 (m, 2H), 3.67-3.63 (m, 4H), 3.94-3.90(m, 2H), 3.03-2.99 (m, 2H), 2.37-2.32 (m, 6H), 2.27 (s, 6H). The preparation method was similar to that of Example 7, except that acryloyl chloride was replaced with 3-N,N-dimethylaminomethacryloyl chloride. The final product was the title compound CXM-08. 1 H-NMR (400 MHz, CDC1 3 ) : δ 8.36 (s, IH), 8.14-8.10 (m, 2H), 7.51-7.48 (m, IH), 7.35-7.31 (m, IH), 7.12-7.08 (m, 2H), 6.84-6.78 (m, 3H), 6.54-6.50 (m, IH), 6.40-6.37 (d, J =2.5 Hz, IH), 6.29-6.26 (d, J =2.5 Hz, IH ), 4.99-4.96 (m, 2H), 3.67-3.63 (m, 4H), 3.94-3.90 (m, 2H), 3.03-2.99 (m, 2H), 2.37-2.32 (m, 6H), 2.27 (s , 6H).
实施例 9 制备^) -N-{7-[( -四氢呋喃 -3-基氧基 ]-4-[ l-(3-氟-苄基) -IH-吲唑 -5-基 胺基] -喹唑啉 -6-基}-4-二甲基胺基-丁 -2-烯酰胺 (化合物 CXM-09) Example 9 Preparation of ^)-N-{7-[(-tetrahydrofuran-3-yloxy]-4-[ l-(3-fluoro-benzyl)-IH-indazol-5-ylamino] - Quinazoline-6-yl}-4-dimethylamino-but-2-enamide (Compound CXM-09)
Figure imgf000016_0001
Figure imgf000016_0001
15 CXM-09  15 CXM-09
冰浴下将化合物 13(430.1g, lmol)溶解于二氯甲垸 (2L)中， 加入 ( -四氢呋 喃基 -3-醇 (105.7g, 1.2mol)， 三苯基磷 (280g, lmol) , 偶氮二羧酸二乙酯 (261.2g, 1.5mol) , 室温搅拌 10h， 反应液过滤， 用二氯甲垸 (1.0L)萃取 3次， 有机相减 压蒸去溶剂， 减压干燥， 得 7-[(R)-四氢呋喃 -3-基氧基] -N-[ l-(3-氟 -苄基 )-1^-吲 唑 -5-基 ]-6-硝基-喹唑啉 -4-胺 (；化合物 14)。 Compound 13 (430.1 g, 1 mol) was dissolved in dichloromethane (2 L) under ice-cooling, (-tetrahydrofuranyl-3-ol (105.7 g, 1.2 mol), triphenylphosphine (280 g, 1 mol), Diethyl azodicarboxylate (261.2g, 1.5mol), stirred at room temperature for 10h, the reaction mixture was filtered, extracted with dichloromethane (1.0L) three times, the organic phase was evaporated under reduced pressure and dried under reduced pressure. 7-[(R)-Tetrahydrofuran-3-yloxy]-N-[ l-(3-fluoro-benzyl)-1^-indazol-5-yl]- 6 -nitro-quinazoline- 4-amine (; compound 14).
参考实施例 2， 化合物 14 经硝基还原反应得 7-[0¾-四氢呋喃 -3-基氧 基] -Λ 4-[ 1-(3-氟-苄基) 吲唑 -5-基] -喹唑啉 -4,6-二胺 (化合物 15)， 再与 3-N,N- 二甲胺基甲基丙烯酰氯进行酰化反应， 得标题化合物 CXM-09。 1H-NMR(400 MHz, CDC13) : δ 8.37 (s, IH), 8.15-8.11 (m, 2H), 7.52-7.47 (m, IH), 7.36-7.32 (m, IH), 7.12-7.05 (m, 2H), 6.83-6.78 (m, 3H), 6.75-6.70 (ά, J =2.5 Hz, IH), 6.54-6.50 (m, IH), 6.41-6.38 (d, J =2.5 Hz, IH), 5.00-4.97 (m, 2H), 4.26-4.22 (m, IH), 4.05-4.01 (m, 2H), 3.80-3.70 (m, 2H), 3.03-2.99 (m, 2H), 2.36-2.34 (m, IH), 2.27 (s, 6H), 2.11-2.09 (m, IH). Reference Example 2, Compound 14 was subjected to nitro reduction to give 7-[03⁄4-tetrahydrofuran-3-yloxy]-indole 4-[1-(3-fluoro-benzyl)oxazol-5-yl]-quin The oxazolidine-4,6-diamine (Compound 15) was acylated with 3-N,N-dimethylaminomethyl methacrylate chloride to give the title compound CXM-09. 1 H-NMR (400 MHz, CDC1 3 ) : δ 8.37 (s, IH), 8.15-8.11 (m, 2H), 7.52-7.47 (m, IH), 7.36-7.32 (m, IH), 7.12-7.05 (m, 2H), 6.83-6.78 (m, 3H), 6.75-6.70 (ά, J =2.5 Hz, IH), 6.54-6.50 (m, IH), 6.41-6.38 (d, J =2.5 Hz, IH ), 5.00-4.97 (m, 2H), 4.26-4.22 (m, IH), 4.05-4.01 (m, 2H), 3.80-3.70 (m, 2H), 3.03-2.99 (m, 2H), 2.36-2.34 (m, IH), 2.27 (s, 6H), 2.11-2.09 (m, IH).
实施例 10 制备^) -N- {4-[l-(3-氟-苄基) - IH-吲唑 -5-基胺基 ]-7- (腈基) -喹唑啉 -6- 基}-4-二甲基胺基-丁 -2-烯酰胺 （化合物 CXM- 10) Example 10 Preparation of ^)-N-{4-[1-(3-Fluoro-benzyl)-IH-indazol-5-ylamino]-7-(nitrile)-quinazoline-6-yl }-4-Dimethylamino-but-2-enamide (Compound CXM-10)
Figure imgf000017_0001
Figure imgf000017_0001
17 CXM-10  17 CXM-10
化合物 13(430.1g, lmol)、氰化亚铜 (89.6g, lmol)加到 DMF(2L)中， 搅拌回 流反应 20h，反应液用硅藻土过滤，滤液中加入 2N盐酸 (0.2L) ,用二氯甲垸 (0.5L) 萃取 3 次， 分离有机相， 减压蒸去溶剂， 减压干燥， 得 4-[1-(3-氟 -节基 )-1^- 吲唑 -5-基胺基 ]-6-硝基-喹唑啉 -7-乙腈 (化合物 16)。  Compound 13 (430.1 g, 1 mol), cuprous cyanide (89.6 g, 1 mol) was added to DMF (2 L), and the mixture was stirred and refluxed for 20 h. The reaction mixture was filtered over Celite, and 2N hydrochloric acid (0.2 L) was added to the filtrate. It was extracted three times with dichloromethane (0.5 L), and the organic phase was separated. The solvent was evaporated under reduced pressure and dried under reduced pressure to give 4-[1-(3-fluoro-pyryl)-1^-carbazole-5- Amino]-6-nitro-quinazolin-7-acetonitrile (compound 16).
参考实施例 2， 化合物 16经过硝基还原反应， 得到 4-[1-(3-氟 -苄基 吲唑 -5-基胺基 ]-6-氨基-喹唑啉 -7-乙腈 (化合物 17)，再与 3-N,N-二甲氨基甲基丙 烯酰氯进行酰化反应， 得标题化合物 CXM-10。 ^-NMRWOO MHz, CDC13) : δ 8.40-8.36 (m, 2Η), 8.24-8.20 (m, IH), 8.16-8.14 (m, IH), 7.37-7.33 (m, IH), 7.12-7.05 (m, 2H), 6.83-6.77 (m, 3H), 6.74-6.70 (d, / =2.5 Hz, IH), 6.53-6.50 (m, IH), 6.40-6.37 (d, J =2.5 Hz, IH), 4.99-4.96 (m, 2H), 3.03-2.99 (m, 2H), 2.27 (s, 6H). Referring to Example 2, Compound 16 was subjected to a nitro reduction reaction to give 4-[1-(3-fluoro-benzyloxazol-5-ylamino)-6-amino-quinazolin-7-acetonitrile (Compound 17 And acylation with 3-N,N-dimethylaminomethacryloyl chloride to give the title compound CXM-10. ^-NMRWOO MHz, CDC1 3 : δ 8.40-8.36 (m, 2 Η), 8.24- 8.20 (m, IH), 8.16-8.14 (m, IH), 7.37-7.33 (m, IH), 7.12-7.05 (m, 2H), 6.83-6.77 (m, 3H), 6.74-6.70 (d, / =2.5 Hz, IH), 6.53-6.50 (m, IH), 6.40-6.37 (d, J = 2.5 Hz, IH), 4.99-4.96 (m, 2H), 3.03-2.99 (m, 2H), 2.27 ( s, 6H).
实施例 11 制备^) -N- {7- [(四氢 -2H-吡喃 -4-基) -甲硫基] -4-[l-(3-氟-苄基) -IH-吲 唑 -5-基胺基] -喹唑啉 -6-基}-4-二甲基胺基-丁 -2-烯酰胺 (化合物 CXM-11) Example 11 Preparation of ^)-N-{7-[(tetrahydro-2H-pyran-4-yl)-methylthio]-4-[l-(3-fluoro-benzyl)-IH-carbazole -5-ylamino]-quinazolin-6-yl}-4-dimethylamino-but-2-enamide (compound CXM-11)
Figure imgf000018_0001
Figure imgf000018_0001
4- (溴甲基) -四氢 -2H-吡喃 (177.9g, lmol)与硫脲 (190g, 2.5mol)回流反应过 夜， 反应液减压浓缩得固体浓缩物， 加入到 30%NaOH水溶液中 2.37L )回流反 应 2h， 然后调节 pH至 3。 混合溶液用乙酸乙酯 (0.5L)萃取 3次， 分离有机相， 无水硫酸钠干燥，浓缩，得淡黄色油状物 4- (硫甲基)四氢 -2H-吡喃 (化合物 18)。  4-(Bromomethyl)-tetrahydro-2H-pyran (177.9 g, 1 mol) was reacted with thiourea (190 g, 2.5 mol) under reflux overnight. The reaction mixture was concentrated under reduced pressure to give a solid concentrate. Medium 2.37 L) reflux reaction for 2 h, then adjust the pH to 3. The mixed solution was extracted with EtOAc (EtOAc) (EtOAc) (EtOAc)
冰盐浴下将钠氢 (96g, 4mol)缓慢加入到化合物 13(430.1g, lmol)的四氢呋 喃 (2L)溶液中， 室温搅拌 0.5h， 然后加入化合物 18(396.2g, 3mol), 室温搅拌 4h。 反应液加入纯化水萃灭， 然后用乙酸乙酯 (0.5L)萃取 3次， 分离有机相， 减压蒸去溶剂，减压干燥，得黄色固体 7- [(四氢 -2H-吡喃 -4-基) -甲硫基] -N-[l-(3- 氟-苄基：) 吲唑 -5-基] -6-硝基-喹唑啉 -4-胺 (化合物 19)。  Sodium hydrogen (96 g, 4 mol) was slowly added to a solution of the compound 13 (430.1 g, 1 mol) in tetrahydrofuran (2 L), and stirred at room temperature for 0.5 h, then compound 18 (396.2 g, 3 mol) was added and stirred at room temperature for 4 h . The reaction mixture was extracted with EtOAc (EtOAc) (EtOAc m. 4-yl)-methylthio]-N-[l-(3-fluoro-benzyl:)oxazol-5-yl]-6-nitro-quinazolin-4-amine (Compound 19).
参考实施例 2， 化合物 19经硝基还原反应， 得 7- [(四氢 -2H-吡喃 -4-基) -甲 硫基] -Λ^Ι-Ρ-氟-苄基) 吲唑 -5-基]-喹唑啉 -4,6-二胺 (化合物 20)， 再与 3_W-二甲氨基甲基丙烯酰氯进行酰化反应， 得标题化合物 CXM-11。 Referring to Example 2, compound 19 was subjected to a nitro reduction reaction to give 7-[(tetrahydro-2H-pyran-4-yl)-methylthio]-indole-oxime-fluoro-benzyl)carbazole- 5-Amino]-quinazoline-4,6-diamine (Compound 20), acylation with 3_W-dimethylaminomethacryloyl chloride to give the title compound CXM-11.
1H-NMR(400 MHz, CDC13) : δ 8.38 (s, 1H), 8.14-8.12 (m, 2H), 7.91-7.87 (m, 1H), 7.35-7.32 (m, 1H), 7.13-7.05 (m, 2H), 6.83-6.77 (m, 3H), 6.74-6.70 (d, J =2.5 Hz, 1H), 6.53-6.50 (m, 1H), 6.40-6.36 (d, J =2.5 Hz, 1H), 4.99-4.96 (m, 2H), 3.65-3.62 (m, 2H), 3.55-3.53 (m, 2H), 3.05-3.03 (m, 2H), 2.73-2.70 (m, 2H), 2.26 (s, 6H), 1.87-1.85 (m, 1H), 1.68-1.64 (m, 2H), 1.43-1.40 (m, 2H). 1 H-NMR (400 MHz, CDC1 3 ) : δ 8.38 (s, 1H), 8.14-8.12 (m, 2H), 7.91-7.87 (m, 1H), 7.35-7.32 (m, 1H), 7.13-7.05 (m, 2H), 6.83-6.77 (m, 3H), 6.74-6.70 (d, J = 2.5 Hz, 1H), 6.53-6.50 (m, 1H), 6.40-6.36 (d, J = 2.5 Hz, 1H ), 4.99-4.96 (m, 2H), 3.65-3.62 (m, 2H), 3.55-3.53 (m, 2H), 3.05-3.03 (m, 2H), 2.73-2.70 (m, 2H), 2.26 (s , 6H), 1.87-1.85 (m, 1H), 1.68-1.64 (m, 2H), 1.43-1.40 (m, 2H).
实施例 12 制备^) -N-{4-[l-(3-氟-苄基) 吲唑 -5-基胺基] -喹唑啉 -6-基 -}-4-二 甲基胺基-丁 -2-烯酰胺 (化合物 CXM-12) Example 12 Preparation of ^)-N-{4-[1-(3-Fluoro-benzyl)oxazol-5-ylamino]-quinazolin-6-yl-}-4-dimethylamino -but-2-enamide (compound CXM-12)
制备方法类似实施例 1， 不同的是将丙烯酰氯替换为 3-N,N-二甲氨基甲基 丙烯酰氯。终产物为标题化合物 CXM-12。 1H-NMR(400 MHz, CDC13) : δ 8.36 (s, IH), 8.22-8.21 (m, 2H), 8.14-8.10 (m, IH), 7.99-7.96 (m, IH), 7.35-7.32 (m, IH), 7.12-7.05 (m, 2H), 6.84-6.78 (m, 3H), 6.75-6.70 (ά, J =2.5 Hz, IH), 6.38-6.34 (d, / =2.5 Hz, IH), 5.01-4.99 (m, 2H), 3.05-3.03 (m, 2H), 2.27 (s, 6H). The preparation method was similar to that of Example 1, except that acryloyl chloride was replaced with 3-N,N-dimethylaminomethacryloyl chloride. The final product was the title compound CXM-12. 1 H-NMR (400 MHz, CDC1 3 ) : δ 8.36 (s, IH), 8.22-8.21 (m, 2H), 8.14-8.10 (m, IH), 7.99-7.96 (m, IH), 7.35-7.32 (m, IH), 7.12-7.05 (m, 2H), 6.84-6.78 (m, 3H), 6.75-6.70 (ά, J =2.5 Hz, IH), 6.38-6.34 (d, / =2.5 Hz, IH ), 5.01-4.99 (m, 2H), 3.05-3.03 (m, 2H), 2.27 (s, 6H).
实施例 13 制备 N- {4-[ l-(3-氟-苄基：) -IH-吲唑 -5-基胺基 ]-7-甲氧基-喹唑啉 -6-基 -} -丙烯酰胺 (化合物 CXM-1 Example 13 Preparation of N-{4-[1-(3-fluoro-benzyl:)-IH-indazol-5-ylamino]-7-methoxy-quinazolin-6-yl-} Acrylamide (compound CXM-1
Figure imgf000019_0001
Figure imgf000019_0001
制备方法类似实施例 2， 不同的是将 3-N,N-二甲氨基甲基丙烯酰氯替换为 丙烯酰氯。 终产物为标题化合物 CXM-13。 ^-NMR^OO MHz , CDC13) : δ 8.38-8.35 (s, IH), 8.14-8.11 (m, 2H), 7.51-7.46 (m, IH), 7.35-7.32 (m, IH), 7.11-7.04 (m, 2H), 6.84-6.78 (m, 3H), 6.54-6.51 (m, IH), 6.48-6.45 (dd, J =2.5 Hz, 8.9 Hz, IH), 6.17-6.14 (d, / = 5.8 Hz, IH), 5.71-5.68 (t, / = 5.8 Hz, IH), 4.99-4.96 (m, 2H), 3.73 (s, 3H). The preparation method was similar to that of Example 2, except that 3-N,N-dimethylaminomethacryloyl chloride was replaced with acryloyl chloride. The final product was the title compound CXM-13. ^-NMR^OO MHz , CDC1 3 ) : δ 8.38-8.35 (s, IH), 8.14-8.11 (m, 2H), 7.51-7.46 (m, IH), 7.35-7.32 (m, IH), 7.11- 7.04 (m, 2H), 6.84-6.78 (m, 3H), 6.54-6.51 (m, IH), 6.48-6.45 (dd, J =2.5 Hz, 8.9 Hz, IH), 6.17-6.14 (d, / = 5.8 Hz, IH), 5.71-5.68 (t, / = 5.8 Hz, IH), 4.99-4.96 (m, 2H), 3.73 (s, 3H).
实施例 14 制备 N- {4-[ l-(3-氟-苄基：) -IH-吲唑 -5-基胺基 ]-7-乙氧基-喹唑啉 -6-基 - 丙烯酰胺 (化合物 CXM-14) Example 14 Preparation of N-{4-[1-(3-fluoro-benzyl:)-IH-indazol-5-ylamino]-7-ethoxy-quinazolin-6-yl-acrylamide (Compound CXM-14)
Figure imgf000020_0001
Figure imgf000020_0001
制备方法类似实施例 13， 不同的是将甲醇钠替换为乙醇钠。 终产物为标 题化合物 CXM- M H-NMRWOO MHz, CDC13): 5 8.38 (s, IH), 8.14-8.11 (m, 2H), 7.50-7.48 (m, IH), 7.37-7.32 (m, IH), 7.12-7.05 (m, 2H), 6.83-6.77 (m, 3H), 6.54-6.50 (m, IH), 6.50-6.48 (dd, J =2.5 Hz, 9.0 Hz, IH), 6.18-6.15 (d, J = 5.8 Hz, IH), 5.70-5.68 (t, / = 5.8 Hz, IH), 5.00-4.96 (m, 2H), 3.98-3.94 (m, 2H), 1.33 (t, J = 7.1 Hz, 3H). The preparation method was similar to that of Example 13, except that sodium methoxide was replaced with sodium ethoxide. The final product is the title compound CXM-M H-NMRWOO MHz, CDC1 3 ): 5 8.38 (s, IH), 8.14-8.11 (m, 2H), 7.50-7.48 (m, IH), 7.37-7.32 (m, IH ), 7.12-7.05 (m, 2H), 6.83-6.77 (m, 3H), 6.54-6.50 (m, IH), 6.50-6.48 (dd, J = 2.5 Hz, 9.0 Hz, IH), 6.18-6.15 ( d, J = 5.8 Hz, IH), 5.70-5.68 (t, / = 5.8 Hz, IH), 5.00-4.96 (m, 2H), 3.98-3.94 (m, 2H), 1.33 (t, J = 7.1 Hz , 3H).
实施例 15 制备 N-{7-[0¾-四氢呋喃 -3-基氧基 ]-4-[l-(3-氟-苄基) -IH-吲唑 -5-基 胺基] -喹唑啉 -6-基- } Example 15 Preparation of N-{7-[03⁄4-tetrahydrofuran-3-yloxy]-4-[l-(3-fluoro-benzyl)-IH-indazol-5-ylamino]-quinazoline -6-based- }
Figure imgf000020_0002
Figure imgf000020_0002
制备方法类似实施例 9， 不同的是将 3-N,N-二甲氨基甲基丙烯酰氯替换为 丙烯酰氯。终产物为标题化合物 CXM-15。 1H-NMR(400 MHz, CDC13): δ 8.37 (s, IH), 8.15-8.11 (m, 2H), 7.52-7.47 (m, IH), 7.36-7.32 (m, IH), 7.11-7.05 (m, 2H), 6.83-6.78 (m, 3H), 6.53-6.50 (m, IH), 6.48-6.44 (dd, J =2.5 Hz, 9.0 Hz, IH), 6.17-6.14 (d, J =5.8 Hz, IH), 5.71-5.68 (t, J = 5.8 Hz, IH), 5.00-4.97 (m, 2H), 4.26-4.22 (m, IH), 4.05-4.01 (m, 2H), 3.78-3.70 (m, 2H), 2.37-2.34 (m, IH), 2.11-2.08 (m, IH). The preparation method was similar to that of Example 9, except that 3-N,N-dimethylaminomethacryloyl chloride was replaced with acryloyl chloride. The final product was the title compound CXM-15. 1 H-NMR (400 MHz, CDC1 3 ): δ 8.37 (s, IH), 8.15-8.11 (m, 2H), 7.52-7.47 (m, IH), 7.36-7.32 (m, IH), 7.11-7.05 (m, 2H), 6.83-6.78 (m, 3H), 6.53-6.50 (m, IH), 6.48-6.44 (dd, J = 2.5 Hz, 9.0 Hz, IH), 6.17-6.14 (d, J = 5.8 Hz, IH), 5.71-5.68 (t, J = 5.8 Hz, IH), 5.00-4.97 (m, 2H), 4.26-4.22 (m, IH), 4.05-4.01 (m, 2H), 3.78-3.70 ( m, 2H), 2.37-2.34 (m, IH), 2.11-2.08 (m, IH).
实施例 16 制备 N-{4-[l-(3-氟-苄基) -IH-吲唑 -5-基胺基 ]-7-腈基-喹唑啉 -6-基 丙烯酰胺 (化合物 CXM-16) Example 16 Preparation of N-{4-[1-(3-fluoro-benzyl)-IH-indazol-5-ylamino]-7-cyano-quinazoline-6-yl acrylamide (Compound CXM -16)
Figure imgf000021_0001
Figure imgf000021_0001
制备方法类似实施例 10， 不同的是将 3-N,N-二甲氨基甲基丙烯酰氯替换 为丙烯酰氯。 终产物为标题化合物 CXM-16。 1H-NMR(400 MHz, CDC13) : δ 8.41-8.35 (m, 2H), 8.24-8.21 (m, IH), 8.14-8.12 (m, IH), 7.35-7.31 (m, IH), 7.10-7.05 (m, 2H), 6.83-6.77 (m, 3H), 6.54-6.50 (m, IH), 6.47-6.43 (dd, J =2.5 Hz, 9.0 Hz, IH), 6.17-6.13 (d, J =5.8 Hz, IH), 5.71-5.68 (t, / = 5.8 Hz, IH), 4.99-4.96 (m, 2H). The preparation was similar to Example 10 except that 3-N,N-dimethylaminomethacryloyl chloride was replaced with acryloyl chloride. The final product was the title compound CXM-16. 1 H-NMR (400 MHz, CDC1 3 ) : δ 8.41-8.35 (m, 2H), 8.24-8.21 (m, IH), 8.14-8.12 (m, IH), 7.35-7.31 (m, IH), 7.10 -7.05 (m, 2H), 6.83-6.77 (m, 3H), 6.54-6.50 (m, IH), 6.47-6.43 (dd, J =2.5 Hz, 9.0 Hz, IH), 6.17-6.13 (d, J =5.8 Hz, IH), 5.71-5.68 (t, / = 5.8 Hz, IH), 4.99-4.96 (m, 2H).
实施例 17 制备 N-{7- [(四氢 -2H-吡喃 -4-基) -甲硫基] -4-[l-(3-氟-苄基) -IH-吲唑 -5-基胺基] -喹唑啉 -6- - } -丙烯酰胺 (化合物 CXM-17) Example 17 Preparation of N-{7-[(tetrahydro-2H-pyran-4-yl)-methylthio]-4-[l-(3-fluoro-benzyl)-IH-indazole-5- Amino]]quinazoline-6--}-acrylamide (compound CXM-17)
Figure imgf000021_0002
Figure imgf000021_0002
制备方法类似实施例 11， 不同的是将 3-N,N-二甲氨基甲基丙烯酰氯替换 为丙烯酰氯。终产物为标题化合物 CXM-17。 1H-NMR(400 MHz, CDC13): δ 8.37 (s, IH), 8.15-8.11 (m, 2H), 7.91-7.88 (m, IH), 7.36-7.32 (m, IH), 7.12-7.05 (m, 2H), 6.84-6.76 (m, 3H), 6.54-6.50 (m, IH), 6.48-6.43 (dd, J =2.5 Hz, 9.0 Hz, IH), 6.18-6.14 (d, J =5.8 Hz, IH), 5.70-5.67 (t, J = 5.8 Hz, IH), 5.00-4.96 (m, 2H)， 3.66-3.62 (m, 2H), 3.56-3.53 (m, 2H), 2.74-2.70 (m, 2H), 1.88-1.85 (m, IH), 1.68-1.65 (m, 2H), 1.44-1.40 (m, 2H). The preparation method was similar to that of Example 11, except that 3-N,N-dimethylaminomethacryloyl chloride was replaced with acryloyl chloride. The final product was the title compound CXM-17. 1 H-NMR (400 MHz, CDC1 3 ): δ 8.37 (s, IH), 8.15-8.11 (m, 2H), 7.91-7.88 (m, IH), 7.36-7.32 (m, IH), 7.12-7.05 (m, 2H), 6.84-6.76 (m, 3H), 6.54-6.50 (m, IH), 6.48-6.43 (dd, J = 2.5 Hz, 9.0 Hz, IH), 6.18-6.14 (d, J = 5.8 Hz, IH), 5.70-5.67 (t, J = 5.8 Hz, IH), 5.00-4.96 (m, 2H), 3.66-3.62 (m, 2H), 3.56-3.53 (m, 2H), 2.74-2.70 ( m, 2H), 1.88-1.85 (m, IH), 1.68-1.65 (m, 2H), 1.44-1.40 (m, 2H).
实施例 18 制备 N-{4-[l-(3-氟-苄基) -IH-吲唑 -5-基胺基] -喹唑啉 -7-基+丙烯酰 胺 (化合物 CXM-18) Example 18 Preparation of N-{4-[1-(3-fluoro-benzyl)-IH-indazol-5-ylamino]-quinazolin-7-yl+acrylamide (Compound CXM-18)
Figure imgf000022_0001
Figure imgf000022_0001
制备方法类似实施例 1，不同的是以 7-硝基喹唑啉酮代替 6-硝基喹唑啉酮。 终产物为标题化合物 CXM-18。 质谱 (ESI+): m/z=438, 439 [M+H] +。 The preparation method was similar to that of Example 1, except that 7-nitroquinazolinone was replaced by 7-nitroquinazolinone. The final product was the title compound CXM-18. Mass Spectrum (ESI + ): m/z = 438, 437 [M+H] + .
实施例 19 制备 (£)-N- {4-[l-(3-氟-苄基) 吲唑 -5-基胺基 ]-6-甲氧基-喹唑啉 -7- 基- }—4-二甲基胺基-丁 -2-烯酰胺化合物 CXM- 19) Example 19 Preparation of (£)-N- {4-[l-(3-fluoro-benzyl)oxazol-5-ylamino]-6-methoxy-quinazolin-7-yl- }- 4-dimethylamino-but-2-enamide compound CXM- 19)
Figure imgf000022_0002
Figure imgf000022_0002
制备方法类似实施例 2， 不同的是以 2-氨基 -5-氯苯甲酸代替 2-氨基 -4-氯苯 甲酸作为起始物料。 终产物为标题化合物 CXM- 19。 质谱 (ESI+): m/z=525, 526 [M+H The preparation method was similar to that of Example 2 except that 2-amino-5-chlorobenzoic acid was used instead of 2-amino-4-chlorobenzoic acid as a starting material. The final product was the title compound CXM-19. Mass Spectrum (ESI + ): m/z = 525, 526 [M+H
实施例 20 制备 (£)-N- {4-[l-(3-氟-苄基) 吲唑 -5-基胺基 ]-6-乙氧基-喹唑啉 -7- 基- }—4-二甲基胺基-丁 -2-烯酰胺 (化合物 CXM-20 Example 20 Preparation of (£)-N- {4-[l-(3-fluoro-benzyl)oxazol-5-ylamino]-6-ethoxy-quinazolin-7-yl- }- 4-dimethylamino-but-2-enamide (compound CXM-20
Figure imgf000022_0003
Figure imgf000022_0003
制备方法类似实施例 19， 不同的是将甲醇钠替换为乙醇钠。 终产物为标 题化合物 CXM-20。 质谱 (ESI+): m/z=539, 540 [M+H] +。 The preparation method was similar to that of Example 19, except that sodium methoxide was replaced with sodium ethoxide. The final product was the title compound CXM-20. Mass Spectrum (ESI + ): m/z = 539, 540 [M+H] + .
实施例 21 制备 (£)-N- {4-[ l-(3-氟-苄基) 吲唑 -5-基胺基 ]-6-(2-甲氧基乙氧 Example 21 Preparation of (£)-N- {4-[ l-(3-fluoro-benzyl)oxazol-5-ylamino]-6-(2-methoxyethoxy
Figure imgf000023_0001
Figure imgf000023_0001
制备方法类似实施例 4和 5， 不同的是将起始原料 4-氯 -6-硝基-喹唑啉 -7- 基-乙酸酯替换为 4-氯 -7-硝基-喹唑啉 -6-基-乙酸酯。 终产物为标题化合物 CXM-2K 质谱 (ESI+): m/z=569, 570 [M+H] +。  The preparation method was similar to that of Examples 4 and 5, except that the starting material 4-chloro-6-nitro-quinazolin-7-yl-acetate was replaced with 4-chloro-7-nitro-quinazoline. -6-yl-acetate. The title product was obtained from the title compound C.M.
实施例 22 制备 N-{4-[l-(3-氟 -苄基 吲唑 -5-基胺基 ]-6-(2-甲氧基乙氧基) -喹 唑啉 -7-基- } -丙烯酰 （化合物 CXM-22) Example 22 Preparation of N-{4-[l-(3-fluoro-benzyloxazol-5-ylamino]-6-(2-methoxyethoxy)-quinazolin-7-yl- } -Acryloyl (Compound CXM-22)
Figure imgf000023_0002
Figure imgf000023_0002
制备方法类似实施例 21， 不同的是将 3-N,N-二甲氨基甲基丙烯酰氯替换 为丙烯酰氯。终产物为标题化合物 CXM-22。质谱 (ESI+): m/z=569, 570[M+H] +。 实施例 23 制备 N-{4-[l-(3-氟-苄基) 吲唑 -5-基胺基 ]-6-[3-(4-吗啉代)丙氧 基]—喹唑啉—7—基- } -丙 The preparation was similar to Example 21 except that 3-N,N-dimethylaminomethacryloyl chloride was replaced with acryloyl chloride. The final product was the title compound CXM-22. Mass Spectrum (ESI + ): m/z = 569, 570 [M+H] + . Example 23 Preparation of N-{4-[1-(3-fluoro-benzyl)oxazol-5-ylamino]-6-[3-(4-morpholino)propoxy]-quinazoline —7—基- } -C
Figure imgf000023_0003
Figure imgf000023_0003
制备方法类似实施例 7，不同的是用 4-氯 -7-硝基-喹唑啉 -6-基 -乙酸酯代替 4-氯 -6-硝基-喹唑啉 -7-基-乙酸酯作为起始原料。 终产物为标题化合物 CXM-23 o 质谱 (ESI+): m/z=581, 582 [M+H] +。  The preparation method was similar to that of Example 7, except that 4-chloro-7-nitro-quinazolin-6-yl-acetate was used instead of 4-chloro-6-nitro-quinazolin-7-yl-B. The acid ester serves as a starting material. The title product was obtained as the title compound: EtOAc (EtOAc): m.
实施例 24 制备^) -N-{4-[l-(3-氟-苄基：) 吲唑 -5-基胺基 ]-6-[3-(4-吗啉代)丙 ^基]—喹唑啉 -7-基 }—4-二甲基胺基-丁 -2-烯酰胺 （化合物 CXM-24) Example 24 Preparation of ^)-N-{4-[1-(3-Fluoro-benzyl:)oxazol-5-ylamino]-6-[3-(4-morpholino)propene ^基]-quinazolin-7-yl}-4-dimethylamino-but-2-enamide (compound CXM-24)
Figure imgf000024_0001
Figure imgf000024_0001
制备方法类似实施例 8，不同的是用 4-氯 -7-硝基-喹唑啉 -6-基 -乙酸酯代替 4-氯 -6-硝基-喹唑啉 -7-基-乙酸酯作为起始原料。 终产物为标题化合物 CXM-24o 质谱 (ESI+): m/z=638, 639 [M+H]+。  The preparation method was similar to that in Example 8, except that 4-chloro-7-nitro-quinazolin-6-yl-acetate was used instead of 4-chloro-6-nitro-quinazolin-7-yl-B. The acid ester serves as a starting material. The title product was the title compound C.M.s.
实施例 25 制备^) -N-{6-[0¾-四氢呋喃 -3-基氧基 ]-4-[1-(3-氟-苄基) 吲唑 -5- 基胺基] -喹唑啉 -7-基- -4-二甲基胺基-丁 -2-烯酰胺 （化合物 CXM-25) Example 25 Preparation of ^)-N-{6-[03⁄4-tetrahydrofuran-3-yloxy]-4-[1-(3-fluoro-benzyl)oxazol-5-ylamino]-quinazoline -7-yl--4-dimethylamino-but-2-enamide (compound CXM-25)
Figure imgf000024_0002
Figure imgf000024_0002
制备方法类似实施例 9，不同的是用 4-氯 -7-硝基-喹唑啉 -6-基 -乙酸酯代替 4-氯 -6-硝基-喹唑啉 -7-基-乙酸酯作为起始原料。 终产物为标题化合物 CXM-25 o 质谱 (ESI+): m/z=581, 582 [M+H]+。  The preparation method was similar to that of Example 9, except that 4-chloro-7-nitro-quinazolin-6-yl-acetate was used instead of 4-chloro-6-nitro-quinazolin-7-yl-B. The acid ester serves as a starting material. The title product was obtained from the title compound C.M.s. (ESI+): m/z = 581, 582 [M+H]+.
实施例 26 制备^) -N-{4-[ l-(3-氟-苄基:) 吲唑 -5-基胺基 ]-6-腈基-喹唑啉 -7-基 -} -4-二甲基胺 -丁 -2-烯酰胺 （化合物 CXM-26) Example 26 Preparation of ^) -N-{4-[ l-(3-fluoro-benzyl:)oxazol-5-ylamino]-6-cyano-quinazolin-7-yl-} -4 -Dimethylamine-but-2-enamide (Compound CXM-26)
Figure imgf000024_0003
Figure imgf000024_0003
制备方法类似实施例 10， 不同的是用 4-氯 -7-硝基-喹唑啉 -6-基-乙酸酯 ^ 替 4-氯 -6-硝基-喹唑啉 -7-基-乙酸酯作为起始原料。 终产物为标题化合 CXM-26o 质谱 (ESI+): m/z=581, 582 [M+H]+。 The preparation method was similar to that in Example 10 except that 4-chloro-7-nitro-quinazolin-6-yl-acetate was used instead of 4-chloro-6-nitro-quinazolin-7-yl- Acetate is used as a starting material. The final product is the title combination CXM-26o mass spectrum (ESI+): m/z = 581, 582 [M+H]+.
实施例 27制备 ( )-N- {6-「(四氢 -2H-吡喃 -4-基) -甲硫基 4-「 3-氟 -苄基 1H-吲 唑 -5-基胺基]-喹唑啉 -7-基 -4-二甲基胺基-丁 -2-烯酰胺 （化合物 CXM-27) Example 27 Preparation of ( )-N-{6-"(tetrahydro-2H-pyran-4-yl)-methylthio 4-"3-fluoro-benzyl 1H-indazol-5-ylamino] -quinazolin-7-yl-4-dimethylamino-but-2-enamide (compound CXM-27)
Figure imgf000025_0001
Figure imgf000025_0001
制备方法类似实施例 11， 不同的是用 4-氯 -7-硝基-喹唑啉 -6-基-乙酸酯代 替 4-氯 -6-硝基-喹唑啉 -7-基-乙酸酯作为起始原料。 终产物为标题化合物 CXM-27。 质谱 (ESI+): m/z=625, 626 [M+H]+。  The preparation method was similar to that in Example 11, except that 4-chloro-7-nitro-quinazolin-6-yl-acetate was used instead of 4-chloro-6-nitro-quinazolin-7-yl-B. The acid ester serves as a starting material. The final product was the title compound CXM-27. Mass Spectrum (ESI+): m/z = 625, 626 [M+H]+.
实施例 28 制备^) -N- {4-[l-(3-氟-苄基) 吲唑 -5-基胺基] -喹唑啉 -7-基 }-4-二 甲基胺基-丁 -2-烯酰胺 （化合物 CXM-28) Example 28 Preparation of ^)-N-{4-[l-(3-Fluoro-benzyl)oxazol-5-ylamino]-quinazolin-7-yl}-4-dimethylamino- But-2-enamide (compound CXM-28)
Figure imgf000025_0002
Figure imgf000025_0002
制备方法类似实施例 12， 不同的是用 7-硝基喹啉酮代替 6-硝基-喹啉酮作 为起始原料。终产物为标题化合物 CXM-28。质谱 (ESI+): m/z=495, 496 [M+H]+。 实施例 29 制备 N- {4-[l-(3-氟-苄基：) 吲唑 -5-基胺基 ]-6-甲氧基-喹唑啉 -7- 基} -丙烯酰胺 (化合 CXM-29) The preparation method was similar to that of Example 12 except that 7-nitroquinolinone was used instead of 6-nitro-quinolinone as a starting material. The final product was the title compound CXM-28. Mass Spectrum (ESI + ): m/z =495, 495 [M+H] + . Example 29 Preparation of N-{4-[1-(3-fluoro-benzyl:)oxazol-5-ylamino]-6-methoxy-quinazolin-7-yl}-acrylamide (combination) CXM-29)
Figure imgf000025_0003
Figure imgf000025_0003
制备方法类似实施例 13， 不同的是用 2-氨基 -5-氯 -苯甲酸代替 2-氨基 -4- 氯-苯甲酸作为起始原料。终产物为标题化合物 CXM-29。质谱 (ESI+): m/z=468, 469 [M+H]+。 The preparation method was similar to that in Example 13, except that 2-amino-5-chloro-benzoic acid was used instead of 2-amino-4-. Chloro-benzoic acid was used as a starting material. The final product was the title compound CXM-29. Mass Spectrum (ESI + ): m/z = 468, 469 [M+H] + .
实施例 30 制备 N-{4-[l-(3-氟-苄基：) 吲唑 -5-基胺基 ]-6-乙氧基-喹唑啉 -7- 基} -丙烯酰胺 (化合物 -30) Example 30 Preparation of N-{4-[1-(3-fluoro-benzyl:)oxazol-5-ylamino]-6-ethoxy-quinazolin-7-yl}-acrylamide (compound) -30)
Figure imgf000026_0001
Figure imgf000026_0001
制备方法类似实施例 14， 不同的是用 2-氨基 -5-氯 -苯甲酸代替 2-氨基 -4- 氯-苯甲酸作为起始原料。终产物为标题化合物 CXM-30。质谱 (ESI+): m/z=482, 483 [M+H]+。 The preparation method was similar to that of Example 14, except that 2-amino-5-chloro-benzoic acid was used instead of 2-amino-4-chloro-benzoic acid as a starting material. The final product was the title compound CXM-30. Mass Spectrum (ESI + ): m/z = 482, 483 [M+H] + .
^Ml L制备 ΛΜ6-[0¾-四氢呋喃 -3-基氧基 ]-4-[1-(3-氟-苄基) 吲唑 -5-基胺 基]-喹唑啉 -7-基} -丙烯酰胺 （化合物 CXM-31)  ^Ml L Preparation of ΛΜ6-[03⁄4-tetrahydrofuran-3-yloxy]-4-[1-(3-fluoro-benzyl)oxazol-5-ylamino]-quinazolin-7-yl} Acrylamide (Compound CXM-31)
Figure imgf000026_0002
Figure imgf000026_0002
制备方法类似实施例 15， 不同的是用 4-氯 -7-硝基-喹唑啉 -6-基-乙酸酯代 替 4-氯 -6-硝基-喹唑啉 -7-基-乙酸酯作为起始原料。 终产物为标题化合物 CXM-31 o 质谱 (ESI+): m/z=524, 525 [M+H]+。  The preparation method was similar to that in Example 15, except that 4-chloro-7-nitro-quinazolin-6-yl-acetate was used instead of 4-chloro-6-nitro-quinazolin-7-yl-B. The acid ester serves as a starting material. The title product was obtained as the title compound C.M.s.
实施例 32 制备 N-{4-[l-(3-氟-苄基) 吲唑 -5-基胺基 ]-6-腈基-喹唑啉 -7-基 丙烯酰胺 (；化合物 CXM- 2) Example 32 Preparation of N-{4-[1-(3-fluoro-benzyl)oxazol-5-ylamino]-6-cyano-quinazolin-7-ylacrylamide (Compound CXM-2 )
Figure imgf000026_0003
制备方法类似实施例 16， 不同的是用 4-氯 -7-硝基-喹唑啉 -6-基-乙酸酯代 替 4-氯 -6-硝基-喹唑啉 -7-基-乙酸酯作为起始原料。 终产物为标题化合物 CXM-32o 质谱 (ESI+): m/z=463, 464 [M+H]+。
Figure imgf000026_0003
The preparation method was similar to that in Example 16, except that 4-chloro-7-nitro-quinazolin-6-yl-acetate was used instead of 4-chloro-6-nitro-quinazolin-7-yl-B. The acid ester serves as a starting material. The title product was obtained from the title compound CXM-32o (ESI+): m/z = 463, 464 [M+H]+.
实施例 33 制备 N-{6- [(四氢 -2H-吡喃 -4基) -甲硫基] -4-[1-(3-氟-苄基) 吲唑 -5-基胺基]-喹唑啉 -7-基} - Example 33 Preparation of N-{6-[(tetrahydro-2H-pyran-4-yl)-methylthio]-4-[1-(3-fluoro-benzyl)oxazol-5-ylamino] -quinazoline-7-yl} -
Figure imgf000027_0001
Figure imgf000027_0001
操作步骤同实施例 17， 不同的是用 4-氯 -7-硝基-喹唑啉 -6-基 -乙酸酯代替 4-氯 -6-硝基-喹唑啉 -7-基-乙酸酯作为起始原料。 终产物为标题化合物 CXM-33 o 质谱 (ESI+): m/z=568, 569 [M+H]+。  The procedure was the same as in Example 17, except that 4-chloro-7-nitro-quinazolin-6-yl-acetate was used instead of 4-chloro-6-nitro-quinazolin-7-yl-B. The acid ester serves as a starting material. The title product was obtained from the title compound: mp.
实施例 34 本发明的化合物对不同酶的抑制活性测试 Example 34 Test of Inhibitory Activity of Compounds of the Invention on Different Enzymes
1、 实验材料：  1. Experimental materials:
本发明的化合物 CXM01~CXM33 ; TK-biotin 底物 （Cat. 62TKOPEC, Cisbio); ATP(Cat. PV3227, Invitrogen); 酶： FGFRl (Cat. PV3146, Invitrogen), PDGFR beta (Cat. P3082, Invitrogen), EGFR(Cat. PV3872, Invitrogen), KDR(Cat. K2643, Sigma), HER2(Cat. PV3366, Invitrogen)； Streptavidin-XL665 (Cat. 62TKOPEC, Cisbio); TK antibody europium cryptate (Cat.62TKOPEC, Cisbio); HTRF kinEASE TK kit (Cat.62TKOPEC, Cisbio)； ENVISION 检测仪： Perkinelmer (Cat. 2104-0010, PerkinElmer)。 Compounds of the invention CXM01~CXM33 ; TK-biotin substrate (Cat. 62TKOPEC, Cisbio); ATP (Cat. PV3227, Invitrogen); Enzyme: FGFRl (Cat. PV3146, Invitrogen), PDGFR beta (Cat. P3082, Invitrogen) , EGFR (Cat. PV3872, Invitrogen), KDR (Cat. K2643, Sigma), HER2 (Cat. PV3366, Invitrogen); Streptavidin-XL665 (Cat. 62TKOPEC, Cisbio); TK antibody europium cryptate (Cat.62TKOPEC, Cisbio) HTRF kinEASE TK kit (Cat.62TKOPEC, Cisbio); ENVISION detector: Perkinelmer (Cat. 2104-0010, PerkinElmer).
2、 实验方法：  2. Experimental method:
应用 HTRF kinEASE TK kit检测本发明 33个化合物对 FGFRl、 PDGFR, Using the HTRF kinEASE TK kit to detect 33 compounds of the invention against FGFR1, PDGFR,
KDR、 EGFR和 HER2酶的抑制活性。 Inhibitory activity of KDR, EGFR and HER2 enzymes.
1 ) 试剂配制  1) Reagent preparation
按表 1所示配制 FGFRl、 PDGF、 KDR、 EGFR和 HER2五种激酶试剂。  Five kinase reagents, FGFR1, PDGF, KDR, EGFR and HER2, were formulated as shown in Table 1.
表 1 五种激酶的反应体系各组分及浓度表  Table 1 Five Kinase Reaction System Components and Concentration Table
反应试剂 FGFRl PDGFR β KDR EGFR HER2 酶浓度 1 ng/well 0.2ng/well 2 ng/well 0.2ng/well 2 ng/well 在酶反应步骤  Reagents FGFRl PDGFR β KDR EGFR HER2 Enzyme concentration 1 ng/well 0.2 ng/well 2 ng/well 0.2 ng/well 2 ng/well in the enzyme reaction step
ATP 浓度 10 μΜ 30 μΜ 5 μΜ 920 nM 50 μΜ 的终浓度 (10  ATP concentration 10 μΜ 30 μΜ 5 μΜ 920 nM 50 μΜ final concentration (10
TK-biotin substrate 465 ηΜ 1 μΜ 200 nM 330 nM 300 nM μ¾  TK-biotin substrate 465 ηΜ 1 μΜ 200 nM 330 nM 300 nM μ3⁄4
酶反应时间 25 min 20 min 20 min 30 min 30 min Streptavidin-XL665浓度 在总反应中的 58.13 ηΜ 125 ηΜ 25 ηΜ 20.6 ηΜ 37.5 ηΜEnzyme reaction time 25 min 20 min 20 min 30 min 30 min The concentration of Streptavidin-XL665 is 58.13 ηΜ 125 ηΜ 25 ηΜ 20.6 ηΜ 37.5 ηΜ in the total reaction.
TK Antibody-Cryptate浓度 终浓度 (20 μί) 1 : 100 稀释 1: 100 稀释 1: 100 稀释 1 : 100 稀释 1 : 100 稀释 l xFGFRl Enzymatic Buffer: lmL 1 x Kinase Buffer 中含有 200 μL 5 x Enzyme buffer, 5 μΐ 1M MgCl2, ΙμΙ 1M DTT, 794 dd¾0. TK Antibody-Cryptate concentration final concentration (20 μί) 1 : 100 dilution 1: 100 dilution 1: 100 dilution 1: 100 dilution 1: 100 dilution l xFGFRl Enzymatic Buffer: lmL 1 x Kinase Buffer contains 200 μL 5 x Enzyme buffer, 5 μΐ 1M MgCl 2 , ΙμΙ 1M DTT, 794 dd3⁄40.
1 xPDGFR Enzymatic Buffer: lmL 1 x Kinase Buffer 中含有 200 μL 5><Enzyme buffer, 5 μΐ 1M MgCl2, ΙμΙ 1M DTT, 20 μΐ SEB, 1 1M MnCl2, 773 μΐ ddH20. 1 xPDGFR Enzymatic Buffer: lmL 1 x Kinase Buffer contains 200 μL 5><Enzyme buffer, 5 μΐ 1M MgCl 2 , ΙμΙ 1M DTT, 20 μΐ SEB, 1 1M MnCl 2 , 773 μΐ ddH 2 0.
l x KDR Enzymatic Buffer: lmL 1 x Kinase Buffer 中含有 200 μL 5 x Enzyme buffer, 5 μΐ 1M MgCl2, ΙμΙ 1M DTT, 1 μΐ 1M MnCl2, 793 μΐ dd¾0. Lx KDR Enzymatic Buffer: lmL 1 x Kinase Buffer contains 200 μL 5 x Enzyme buffer, 5 μΐ 1M MgCl 2 , ΙμΙ 1M DTT, 1 μΐ 1M MnCl 2 , 793 μΐ dd3⁄40.
l x EGFR Enzymatic Buffer: lmL 1 x Kinase Buffer 中含有 200 μL 5><Enzyme buffer, 5 μΐ 1M MgCl2, ΙμΙ 1M DTT, 1 μΐ 1M MnCl2, 793 μΐ dd¾0. Lx EGFR Enzymatic Buffer: lmL 1 x Kinase Buffer contains 200 μL 5><Enzyme buffer, 5 μΐ 1M MgCl 2 , ΙμΙ 1M DTT, 1 μΐ 1M MnCl 2 , 793 μΐ dd3⁄40.
l xHER2 Enzymatic Buffer: lmL 1 x Kinase Buffer 中含有 200 μL 5 x Enzyme buffer, 5 1M MgCl2, ΙμΙ 1M DTT, 3.2 μΐ SEB, 1 1M MnCl2, 789.8 ddH20. l xHER2 Enzymatic Buffer: lmL 1 x Kinase Buffer contains 200 μL 5 x Enzyme buffer, 5 1M MgCl 2 , ΙμΙ 1M DTT, 3.2 μΐ SEB, 1 1M MnCl 2 , 789.8 ddH 2 0.
5 Substrate- TK和 ATP工作液  5 Substrate- TK and ATP working fluids
Substrate-TK和 ATP的具体浓度见表 1。 The specific concentrations of Substrate-TK and ATP are shown in Table 1.
用 1 x Kinase Buffer稀释 Substrate-TK和 ATP至反应浓度的 5倍。 Substrate-TK and ATP were diluted to 5 times the reaction concentration with 1 x Kinase Buffer.
5xEnzyme 工作液： 五种激酶浓度优化筛选时所用试剂的浓度见表 1。 用 l xkinase buffer分别配制 5种酶的 5χ酶工作液。  5xEnzyme working solution: The concentration of the reagents used in the five kinase concentration optimization screening is shown in Table 1. Five enzymes of 5 enzymes were prepared using l xkinase buffer.
4xStreptavidin -XL665 工作液： Sa-XL665在反应中的浓度见表 1。用 Detection Buffer配制 4χ Streptavidin -XL665 工作液。  4x Streptavidin - XL665 working solution: Sa-XL665 concentration in the reaction is shown in Table 1. Prepare 4χ Streptavidin -XL665 working solution with Detection Buffer.
4x TK-Antibody-cryptate工作液： 用 Detection Buffer将 TK- Antibody-Cryptate 稀释 100倍作为工作液。 4x TK-Antibody-cryptate working solution: Dilute TK- Antibody-Cryptate 100 times as a working solution with Detection Buffer.
所有试剂按照上述方法配好后， 除酶外， 平衡到室温以后， 开始加样。 a) 首先使用配置好的五种酶的 IX kinase buffer分别配制 2.5%的 DMSO 溶液（DMSO浓度过高会对反应产生影响， 控制 DMSO的终浓度为 1% ) , 然 后用五种酶对应的 2.5%的 DMSO溶液稀释待测化合物， 每个待测物的终浓度 为 2 μΜ和 0.2 μΜ。  After all the reagents were prepared as described above, the enzyme was added to the room temperature after the enzyme was added to the room temperature. a) First prepare a 2.5% DMSO solution using the IX kinase buffers of the five enzymes configured (the DMSO concentration will affect the reaction, control the final concentration of DMSO to 1%), and then use the five enzymes corresponding to 2.5. The test compound was diluted with % DMSO solution, and the final concentration of each analyte was 2 μΜ and 0.2 μΜ.
除对照孔外， 向所有反应孔中加入 4μ1的稀释好的待测化合物溶液， 向对 照孔中加入 4μ1先前配制的五种酶对应的 2.5%的 DMSO溶液。  In addition to the control wells, 4 μl of the diluted test compound solution was added to all wells, and 4 μl of the previously prepared five enzymes corresponding to 2.5% of DMSO solution were added to the control wells.
b ) 向所有反应孔中加入 2μ1 先前配制好的五种酶对应底物浓度的 TK-biotin substrate溶液 （五种酶筛选时底物的用量见表 1 )。  b) Add 2 μl of the previously prepared NK-biotin substrate solution to the substrate at the substrate concentration (see Table 1 for the amount of substrate for the five enzymes).
c) 向除阴性孔外的所有反应孔中加入 2μ1先前配制好的对应浓度的五种 酶溶液（五种酶的用量见表 1 )， 阴性孔用 2μ1五种酶对应 IXkinase buffer补足 体积。 用封板膜封板， 混匀后室温孵育 lOmin, 让化合物和酶充分作用结合。 d)向所有反应孔中加入 2μ1五种酶对应浓度的 ΑΤΡ溶液来启动激酶反应， 其中 PDGFR β和 KDR酶反应时间为 20min， FGFR1酶反应时间为 25min， EGFR和 HER2的酶反应时间为 30min。五种酶筛选时对应的 ATP浓度和反应 时间见表 1。 c) Add 2μ1 of the previously prepared five enzyme solutions to all the wells except the negative wells (see Table 1 for the amount of the five enzymes), and the negative wells with 2μ1 of the five enzymes for the IXkinase buffer. Volume. The plate was sealed with a sealing plate, and the mixture was incubated at room temperature for 10 min to allow the compound and the enzyme to fully bind. d) The kinase reaction was initiated by adding 2 μl of five enzymes corresponding to the concentration of ruthenium solution to all wells, wherein the PDGFR β and KDR enzyme reaction time was 20 min, the FGFR1 enzyme reaction time was 25 min, and the EGFR and HER 2 enzymatic reaction time was 30 min. The corresponding ATP concentration and reaction time for the five enzyme screenings are shown in Table 1.
e ) 在激酶反应结束前 5min 开始配制五种酶检测液。 使用试剂盒中的 detection buffer 分别配制五种酶对应浓度 的 Streptavidin-XL665 和 TK antibody europium cryptate(l : 100) 检测液。 五种酶筛选时对应的检测试剂浓度 见表 1。  e) Prepare five enzyme assays 5 minutes before the end of the kinase reaction. Five concentrations of Streptavidin-XL665 and TK antibody europium cryptate (l: 100) were prepared using the detection buffer in the kit. The corresponding detection reagent concentrations for the five enzyme screenings are shown in Table 1.
f) 待激酶反应结束后， 向所有反应孔中加入 5μ1 五种酶对应稀释的 f) After the end of the kinase reaction, add 5μ1 to all wells and dilute the corresponding enzymes.
Streptavidin-XL665 溶液， 混匀后立即加入稀释好的 TK antibody europium cryptate检测液。 Streptavidin-XL665 solution, add the diluted TK antibody europium cryptate test solution immediately after mixing.
g)封板混匀， 室温反应 lh后， 用 ENVISION检测仪检测荧光信号（320 nm刺激， 665 nm， 615 nm发射）。 通过全活性孔和背景信号孔计算出每个孔 的抑制率， 复孔取平均值， 同时用专业的画图分析软件 PRISM 5.0对每个待 测化合物进行半数抑制活性 （IC50) 的拟合。  g) The plate was mixed and reacted at room temperature for 1 h, and the fluorescence signal (320 nm stimulation, 665 nm, 615 nm emission) was detected by an ENVISION detector. The inhibition rate of each well was calculated from the fully active wells and the background signal wells, and the duplicate wells were averaged, and the half-inhibitory activity (IC50) of each test compound was fitted using a professional paint analysis software PRISM 5.0.
发射率 （ER)= 665 nm发射信号 / 615 nm发射信号  Emissivity (ER) = 665 nm transmit signal / 615 nm transmit signal
抑制率 = (ER賺 -ER样品) /(ER阳性 -ER阴性) *100%  Inhibition rate = (ER earn - ER sample) / (ER positive - ER negative) * 100%
实验加样流程图如下: The experimental sample flow chart is as follows:
Figure imgf000029_0001
Figure imgf000029_0001
3、 结果与分析 表 2-表 6分别显示本发明的化合物在 2μΜ和 0.2μΜ浓度时对 FGFR1、 PDGFR、 KDR、 EGFR和 HER2酶的抑制率。 3. Results and analysis Tables 2 - 6 show the inhibition rates of the FGFRl, PDGFR, KDR, EGFR and HER2 enzymes of the compounds of the invention at concentrations of 2 μΜ and 0.2 μ, respectively.
表 2本发明的化合物在 2 μΜ和 0.2 μΜ浓度时对 FGFR1的抑制率  Table 2 Inhibition rate of FGFR1 at concentrations of 2 μΜ and 0.2 μΜ of the compounds of the present invention
浓度 2 μΜ 浓度 0.2 μΜ  Concentration 2 μΜ concentration 0.2 μΜ
化合物编号  Compound number
%抑制率 SD %抑制率 SD % inhibition rate SD % inhibition rate SD
CXM-01 37.76 9.62 6.56 7.81CXM-01 37.76 9.62 6.56 7.81
CXM-02 97.12 2.38 65.76 7.58CXM-02 97.12 2.38 65.76 7.58
CXM-03 41.03 0.63 63.00 2.96CXM-03 41.03 0.63 63.00 2.96
CXM-04 50.52 1.37 90.88 0.11CXM-04 50.52 1.37 90.88 0.11
CXM-05 67.79 13.95 82.06 9.41CXM-05 67.79 13.95 82.06 9.41
CXM-06 -5.60 4.41 -7.04 1.92CXM-06 -5.60 4.41 -7.04 1.92
CXM-07 27.76 0.00 -1.76 0.79CXM-07 27.76 0.00 -1.76 0.79
CXM-08 -17.84 1.13 3.20 2.15CXM-08 -17.84 1.13 3.20 2.15
CXM-09 72.65 8.98 95.96 0.32CXM-09 72.65 8.98 95.96 0.32
CXM-10 8.24 11.99 69.12 16.40CXM-10 8.24 11.99 69.12 16.40
CXM-11 4.00 10.07 1.76 3.28CXM-11 4.00 10.07 1.76 3.28
CXM-12 44.92 9.09 76.83 1.16CXM-12 44.92 9.09 76.83 1.16
CXM-13 45.96 13.95 48.88 7.50CXM-13 45.96 13.95 48.88 7.50
CXM-14 33.41 9.72 62.18 7.08CXM-14 33.41 9.72 62.18 7.08
CXM-15 63.23 12.16 76.68 7.50CXM-15 63.23 12.16 76.68 7.50
CXM-16 -39.84 7.81 7.28 7.92CXM-16 -39.84 7.81 7.28 7.92
CXM-17 11.51 7.72 53.66 2.22CXM-17 11.51 7.72 53.66 2.22
CXM-18 50.52 1.59 73.99 0.32CXM-18 50.52 1.59 73.99 0.32
CXM-19 -16.24 4.07 1.20 14.71CXM-19 -16.24 4.07 1.20 14.71
CXM-20 50.37 2.64 85.80 8.98CXM-20 50.37 2.64 85.80 8.98
CXM-21 -6.40 13.24 61.81 2.49CXM-21 -6.40 13.24 61.81 2.49
CXM-22 50.37 2.43 83.86 3.91CXM-22 50.37 2.43 83.86 3.91
CXM-23 32.88 1.80 71.60 13.21CXM-23 32.88 1.80 71.60 13.21
CXM-24 90.21 0.21 85.95 6.24CXM-24 90.21 0.21 85.95 6.24
CXM-25 2.32 5.07 49.10 2.33CXM-25 2.32 5.07 49.10 2.33
CXM-26 55.16 2.43 68.76 2.22CXM-26 55.16 2.43 68.76 2.22
CXM-27 50.07 3.91 80.57 0.11CXM-27 50.07 3.91 80.57 0.11
CXM-28 43.50 11.73 48.21 0.42CXM-28 43.50 11.73 48.21 0.42
CXM-29 22.05 4.23 58.67 14.16 CXM-30 76.01 1.48 91.93 4.33CXM-29 22.05 4.23 58.67 14.16 CXM-30 76.01 1.48 91.93 4.33
CXM-31 49.48 8.35 73.02 0.63CXM-31 49.48 8.35 73.02 0.63
CXM-32 37.97 13.85 65.02 13.42CXM-32 37.97 13.85 65.02 13.42
CXM-33 90.21 0.21 85.95 6.24 阿帕替尼 40.58 0.63 64.35 11.42CXM-33 90.21 0.21 85.95 6.24 Apatinib 40.58 0.63 64.35 11.42
Staurosporine 100.15 1.80 98.88 1.27 从表 2的结果可以看出， 化合物 CXM-02、 CXM-04、 CXM-05、 CXM-09、 CXM-15、 CXM-18、 CXM-20、 CXM-22、 CXM-26、 CXM-27、 CXM-30、 CXM-31、 CXM-33在浓度 2 μΜ和 0.2 μΜ都表现了比阳性对照阿帕替尼较好或相当的活 性。 表 3本发明的化合物在 2 μΜ和 0.2 μΜ浓度时对 PDGFR beta的抑制率 浓度 2 μΜ 浓度 0.2 μΜ Staurosporine 100.15 1.80 98.88 1.27 As can be seen from the results in Table 2, the compounds CXM-02, CXM-04, CXM-05, CXM-09, CXM-15, CXM-18, CXM-20, CXM-22, CXM-26 CXM-27, CXM-30, CXM-31, and CXM-33 showed better or comparable activity than the positive control apatinib at concentrations of 2 μΜ and 0.2 μΜ. Table 3 Inhibition of PDGFR beta at concentrations of 2 μΜ and 0.2 μΜ of the compounds of the present invention Concentration 2 μΜ Concentration 0.2 μΜ
化合物编号  Compound number
%抑制率 SD %抑制率 SD  % inhibition rate SD % inhibition rate SD
CXM-01 10.78 7.30 18.06 2.55 CXM-01 10.78 7.30 18.06 2.55
CXM-02 34.26 2.85 14.71 8.03CXM-02 34.26 2.85 14.71 8.03
CXM-03 33.78 10.19 34.30 0.73CXM-03 33.78 10.19 34.30 0.73
CXM-04 20.53 0.00 11.45 3.37CXM-04 20.53 0.00 11.45 3.37
CXM-05 22.33 6.19 13.26 5.92CXM-05 22.33 6.19 13.26 5.92
CXM-06 -7.17 11.82 19.92 13.06CXM-06 -7.17 11.82 19.92 13.06
CXM-07 -11.97 7.66 10.27 5.84CXM-07 -11.97 7.66 10.27 5.84
CXM-08 -5.16 13.50 5.73 15.03CXM-08 -5.16 13.50 5.73 15.03
CXM-09 35.65 7.55 15.70 0.64CXM-09 35.65 7.55 15.70 0.64
CXM-10 2.68 3.58 11.51 7.74CXM-10 2.68 3.58 11.51 7.74
CXM-11 13.36 0.15 15.12 2.92CXM-11 13.36 0.15 15.12 2.92
CXM-12 26.90 2.82 13.19 6.55CXM-12 26.90 2.82 13.19 6.55
CXM-13 21.75 1.05 9.67 3.94CXM-13 21.75 1.05 9.67 3.94
CXM-14 9.60 6.31 10.66 0.61CXM-14 9.60 6.31 10.66 0.61
CXM-15 17.95 7.83 -1.99 8.01CXM-15 17.95 7.83 -1.99 8.01
CXM-16 -0.52 14.67 13.42 1.97CXM-16 -0.52 14.67 13.42 1.97
CXM-17 15.06 1.40 10.90 7.10CXM-17 15.06 1.40 10.90 7.10
CXM-18 32.11 1.82 21.62 4.64CXM-18 32.11 1.82 21.62 4.64
CXM-19 10.99 0.29 21.52 6.13CXM-19 10.99 0.29 21.52 6.13
CXM-20 46.27 1 19.63 13.83 CXM-21 6.97 1.75 24.15 1.68CXM-20 46.27 1 19.63 13.83 CXM-21 6.97 1.75 24.15 1.68
CXM-22 27.67 0.46 13.71 5.28CXM-22 27.67 0.46 13.71 5.28
CXM-23 29.43 5.96 5.08 10.95CXM-23 29.43 5.96 5.08 10.95
CXM-24 47.23 10.28 29.47 10.10CXM-24 47.23 10.28 29.47 10.10
CXM-25 9.73 0.35 17.47 6.75CXM-25 9.73 0.35 17.47 6.75
CXM-26 13.88 0.09 6.01 2.98CXM-26 13.88 0.09 6.01 2.98
CXM-27 20.07 7.80 -5.64 5.61CXM-27 20.07 7.80 -5.64 5.61
CXM-28 15.18 9.81 3.59 13.23CXM-28 15.18 9.81 3.59 13.23
CXM-29 5.33 9.90 -0.12 2.19CXM-29 5.33 9.90 -0.12 2.19
CXM-30 32.88 1.64 24.90 4.19CXM-30 32.88 1.64 24.90 4.19
CXM-31 35.38 2.28 16.79 14.90CXM-31 35.38 2.28 16.79 14.90
CXM-32 48.88 0.35 16.17 5.08CXM-32 48.88 0.35 16.17 5.08
CXM-33 47.23 10.28 29.47 10.10 阿帕替尼 39.45 4.91 35.71 3.82CXM-33 47.23 10.28 29.47 10.10 Apatinib 39.45 4.91 35.71 3.82
Staurosporine 99.87 0.27 100.32 0.18 从表 3的结果可以看出，化合物 CXM-03、 CXM-24、 CXM-33在浓度 2 μΜ2 μΜ都表现出与阳性对照阿帕替尼相当的活性。 表 4 本发明的化合物在 2 μΜ和 0.2 μΜ浓度时对 KDR的抑制率 Staurosporine 99.87 0.27 100.32 0.18 As can be seen from the results in Table 3, the compounds CXM-03, CXM-24, and CXM-33 exhibited activity comparable to the positive control apatinib at a concentration of 2 μΜ2 μΜ. Table 4 Inhibition rate of KDR in the compounds of the present invention at concentrations of 2 μΜ and 0.2 μΜ
浓度 2 μΜ 浓度 0.2 μΜ  Concentration 2 μΜ concentration 0.2 μΜ
化合物编号  Compound number
%抑制率 SD %抑制率 SD % inhibition rate SD % inhibition rate SD
CXM-01 70.47 1.63 27.94 11.44CXM-01 70.47 1.63 27.94 11.44
CXM-02 62.09 4.05 22.21 5.51CXM-02 62.09 4.05 22.21 5.51
CXM-03 44.39 10.62 43.15 11.96CXM-03 44.39 10.62 43.15 11.96
CXM-04 27.48 4.23 65.45 4.74CXM-04 27.48 4.23 65.45 4.74
CXM-05 44.31 4.95 60.13 1.13CXM-05 44.31 4.95 60.13 1.13
CXM-06 21.69 0.54 7.92 3.21CXM-06 21.69 0.54 7.92 3.21
CXM-07 -4.92 3.33 -12.71 0.06CXM-07 -4.92 3.33 -12.71 0.06
CXM-08 18.78 2.48 2.61 11.80CXM-08 18.78 2.48 2.61 11.80
CXM-09 68.00 2.16 65.67 6.08CXM-09 68.00 2.16 65.67 6.08
CXM-10 31.19 2.60 18.44 2.36CXM-10 31.19 2.60 18.44 2.36
CXM-11 25.07 6.29 26.10 0.36CXM-11 25.07 6.29 26.10 0.36
CXM-12 36.52 6.49 26.46 1.13CXM-12 36.52 6.49 26.46 1.13
CXM-13 30.25 12.68 18.37 1.65 CXM-14 33.38 1.24 19.39 2.47CXM-13 30.25 12.68 18.37 1.65 CXM-14 33.38 1.24 19.39 2.47
CXM-15 38.34 6.60 41.25 1.86CXM-15 38.34 6.60 41.25 1.86
CXM-16 11.08 16.40 -4.96 6.29CXM-16 11.08 16.40 -4.96 6.29
CXM-17 27.33 5.26 2.48 2.68CXM-17 27.33 5.26 2.48 2.68
CXM-18 23.91 11.13 13.99 2.27CXM-18 23.91 11.13 13.99 2.27
CXM-19 43.00 0.00 25.72 1.63CXM-19 43.00 0.00 25.72 1.63
CXM-20 40.74 14.74 36.73 3.50CXM-20 40.74 14.74 36.73 3.50
CXM-21 12.45 4.30 31.66 10.05CXM-21 12.45 4.30 31.66 10.05
CXM-22 31.27 4.64 68.15 6.08CXM-22 31.27 4.64 68.15 6.08
CXM-23 61.95 1 29.81 14.33CXM-23 61.95 1 29.81 14.33
CXM-24 93.29 2.47 50.00 3.09CXM-24 93.29 2.47 50.00 3.09
CXM-25 37.97 12.06 34.55 0.62CXM-25 37.97 12.06 34.55 0.62
CXM-26 6.27 2.47 23.10 13.71CXM-26 6.27 2.47 23.10 13.71
CXM-27 3.50 3.71 18.37 0.21CXM-27 3.50 3.71 18.37 0.21
CXM-28 14.36 8.76 16.84 14.12CXM-28 14.36 8.76 16.84 14.12
CXM-29 8.45 9.28 -2.33 0.41CXM-29 8.45 9.28 -2.33 0.41
CXM-30 76.09 7.22 65.38 5.26CXM-30 76.09 7.22 65.38 5.26
CXM-31 63.41 1 47.74 2.16CXM-31 63.41 1 47.74 2.16
CXM-32 51.68 0.72 57.65 8.14CXM-32 51.68 0.72 57.65 8.14
CXM-33 34.10 6.35 32.69 1.94 阿帕替尼 49.20 4.43 21.94 12.27CXM-33 34.10 6.35 32.69 1.94 Apatinib 49.20 4.43 21.94 12.27
Staurosporine 100.15 0.21 99.42 0.21 从表 4的结果可以看出， 化合物 CXM-01、 CXM-02、 CXM-09、 CXM-23、 CXM-24, CXM-30, CXM-31、 CXM-32在浓度 2 μΜ和 0.2 μΜ都表现了比阳 性对照阿帕替尼较好或相当的活性。 表 5 本发明的化合物在 2 μΜ和 0.2 μΜ浓度时对 EGFR抑制率 Staurosporine 100.15 0.21 99.42 0.21 As can be seen from the results in Table 4, the compounds CXM-01, CXM-02, CXM-09, CXM-23, CXM-24, CXM-30, CXM-31, CXM-32 at a concentration of 2 μΜ Both 0.2 μΜ showed better or comparable activity than the positive control apatinib. Table 5 EGFR inhibition rate of compounds of the invention at concentrations of 2 μΜ and 0.2 μΜ
浓度 2 μΜ 浓度 0.2 μΜ  Concentration 2 μΜ concentration 0.2 μΜ
化合物编号  Compound number
%抑制率 SD %抑制率 SD  % inhibition rate SD % inhibition rate SD
CXM-01 100.40 0.21 99.64 0.43 CXM-01 100.40 0.21 99.64 0.43
CXM-02 99.94 0.34 99.70 0.52CXM-02 99.94 0.34 99.70 0.52
CXM-03 100.32 0.17 100.09 0.50CXM-03 100.32 0.17 100.09 0.50
CXM-04 99.74 0.33 101.02 0.50CXM-04 99.74 0.33 101.02 0.50
CXM-05 100.15 0.25 100.15 0.08 CXM-06 100.12 0.26 99.88 0.09CXM-05 100.15 0.25 100.15 0.08 CXM-06 100.12 0.26 99.88 0.09
CXM-07 99.91 0.56 99.67 0.21CXM-07 99.91 0.56 99.67 0.21
CXM-08 100.58 0.47 100.00 0.09CXM-08 100.58 0.47 100.00 0.09
CXM-09 98.63 1.07 99.91 1.07CXM-09 98.63 1.07 99.91 1.07
CXM-10 100.30 0.43 99.06 0.21CXM-10 100.30 0.43 99.06 0.21
CXM-11 100.40 0.04 100.06 0.69CXM-11 100.40 0.04 100.06 0.69
CXM-12 101.02 0.17 101.67 0.74CXM-12 101.02 0.17 101.67 0.74
CXM-13 102.31 0.66 99.62 0.17CXM-13 102.31 0.66 99.62 0.17
CXM-14 101.26 0.00 94.30 2.40CXM-14 101.26 0.00 94.30 2.40
CXM-15 100.20 0.50 100.96 0.25CXM-15 100.20 0.50 100.96 0.25
CXM-16 99.73 0.21 99.87 0.17CXM-16 99.73 0.21 99.87 0.17
CXM-17 101.20 0.08 78.93 5.95CXM-17 101.20 0.08 78.93 5.95
CXM-18 99.68 0.58 100.79 0.17CXM-18 99.68 0.58 100.79 0.17
CXM-19 99.67 0.21 63.31 2.54CXM-19 99.67 0.21 63.31 2.54
CXM-20 100.03 0.08 100.44 0.99CXM-20 100.03 0.08 100.44 0.99
CXM-21 100.21 0.13 96.60 2.49CXM-21 100.21 0.13 96.60 2.49
CXM-22 99.97 0.99 100.73 0.41CXM-22 99.97 0.99 100.73 0.41
CXM-23 101.37 0.17 99.62 0.66CXM-23 101.37 0.17 99.62 0.66
CXM-24 100.96 0.41 100.44 0.83CXM-24 100.96 0.41 100.44 0.83
CXM-25 101.90 0.08 96.05 0.41CXM-25 101.90 0.08 96.05 0.41
CXM-26 101.55 0.91 97.17 0.17CXM-26 101.55 0.91 97.17 0.17
CXM-27 101.67 0.25 88.28 3.14CXM-27 101.67 0.25 88.28 3.14
CXM-28 101.32 0.25 88.28 1.49CXM-28 101.32 0.25 88.28 1.49
CXM-29 100.03 0.58 71.74 1.90CXM-29 100.03 0.58 71.74 1.90
CXM-30 100.09 0.83 99.15 0.50CXM-30 100.09 0.83 99.15 0.50
CXM-31 99.97 0.83 100.85 1.07CXM-31 99.97 0.83 100.85 1.07
CXM-32 100.20 0.17 99.85 0.99CXM-32 100.20 0.17 99.85 0.99
CXM-33 99.48 0.21 99.42 0.39 阿帕替尼 100.91 0.66 100.20 0.83CXM-33 99.48 0.21 99.42 0.39 Apatinib 100.91 0.66 100.20 0.83
Staurosporine 99.15 0.17 90.67 1.59 从表 5的结果可以看出， 化合物 CXM-01、 CXM-02、 CXM-09, CXM-23、 CXM-24, CXM-30, CXM-31、 CXM-32在浓度 2 μΜ和 0.2 μΜ都表现了比阳 性对照阿帕替尼较好或相当的活性。 表 6 本发明的化合物在 2 μΜ和 0.2 μΜ浓度时对 HER2的抑制率 浓度 2 μΜ 浓度 0.2 μΜ 化合物编号 Staurosporine 99.15 0.17 90.67 1.59 As can be seen from the results in Table 5, the compounds CXM-01, CXM-02, CXM-09, CXM-23, CXM-24, CXM-30, CXM-31, CXM-32 at a concentration of 2 μΜ Both 0.2 μΜ showed better or comparable activity than the positive control apatinib. Table 6 Inhibition rate of HER2 at a concentration of 2 μΜ and 0.2 μΜ of the compound of the present invention 2 μΜ concentration 0.2 μΜ Compound number
%抑制率 SD %抑制率 SD % inhibition rate SD % inhibition rate SD
CXM-01 99.77 0.22 92.64 0.76CXM-01 99.77 0.22 92.64 0.76
CXM-02 98.47 0.25 95.93 0.64CXM-02 98.47 0.25 95.93 0.64
CXM-03 100.06 0.07 95.29 1.82CXM-03 100.06 0.07 95.29 1.82
CXM-04 88.37 3.51 49.98 0.78CXM-04 88.37 3.51 49.98 0.78
CXM-05 99.23 0.10 84.12 1.28CXM-05 99.23 0.10 84.12 1.28
CXM-06 99.55 0.25 71.60 2.63CXM-06 99.55 0.25 71.60 2.63
CXM-07 99.50 0.22 73.27 0.61CXM-07 99.50 0.22 73.27 0.61
CXM-08 99.86 0.10 74.42 0.12CXM-08 99.86 0.10 74.42 0.12
CXM-09 99.99 0.24 97.89 0.64CXM-09 99.99 0.24 97.89 0.64
CXM-10 99.34 0.30 79.58 1.52CXM-10 99.34 0.30 79.58 1.52
CXM-11 99.48 0.15 72.80 0.34CXM-11 99.48 0.15 72.80 0.34
CXM-12 100.52 0.24 99.61 0.03CXM-12 100.52 0.24 99.61 0.03
CXM-13 99.83 0.20 66.39 6.45CXM-13 99.83 0.20 66.39 6.45
CXM-14 96.22 1.92 38.47 1.45CXM-14 96.22 1.92 38.47 1.45
CXM-15 100.04 0.57 95.48 0.81CXM-15 100.04 0.57 95.48 0.81
CXM-16 97.08 0.54 37.06 7.21CXM-16 97.08 0.54 37.06 7.21
CXM-17 98.20 0.07 35.01 6.68CXM-17 98.20 0.07 35.01 6.68
CXM-18 99.94 0.17 95.24 1.08CXM-18 99.94 0.17 95.24 1.08
CXM-19 77.20 0.81 40.12 0.32CXM-19 77.20 0.81 40.12 0.32
CXM-20 99.73 0.47 92.86 1.01CXM-20 99.73 0.47 92.86 1.01
CXM-21 99.30 0.20 62.97 0.76CXM-21 99.30 0.20 62.97 0.76
CXM-22 99.66 0.37 84.51 3.37CXM-22 99.66 0.37 84.51 3.37
CXM-23 99.42 0.30 39.26 17.41CXM-23 99.42 0.30 39.26 17.41
CXM-24 100.23 0.03 99.61 0.30CXM-24 100.23 0.03 99.61 0.30
CXM-25 96.58 2.97 30.62 10.46CXM-25 96.58 2.97 30.62 10.46
CXM-26 96.94 1.32 33.70 15.15CXM-26 96.94 1.32 33.70 15.15
CXM-27 95.55 0.17 23.35 1.38CXM-27 95.55 0.17 23.35 1.38
CXM-28 97.92 1.35 24.13 0.00CXM-28 97.92 1.35 24.13 0.00
CXM-29 94.98 3.48 15.21 5.53CXM-29 94.98 3.48 15.21 5.53
CXM-30 92.21 0.69 78.33 5.80CXM-30 92.21 0.69 78.33 5.80
CXM-31 99.66 0.64 72.67 4.72 CXM-32 99.61 0.03 80.47 1.72CXM-31 99.66 0.64 72.67 4.72 CXM-32 99.61 0.03 80.47 1.72
CXM-33 100.02 0.20 97.89 0.51 阿帕替尼 99.85 0.03 97.39 0.34CXM-33 100.02 0.20 97.89 0.51 apatinib 99.85 0.03 97.39 0.34
Staurosporine 89.55 0.07 54.73 8.63 从表 6的结果可以看出， 化合物 CXM-01、 CXM-02、 CXM-03、 CXM-09、 CXM-12、 CXM-15、 CXM-18、 CXM-20、 CXM-24在浓度 2 μΜ和 0.2 μΜ都 表现了比阳性对照阿帕替尼较好或相当的活性。 Staurosporine 89.55 0.07 54.73 8.63 As can be seen from the results in Table 6, the compounds CXM-01, CXM-02, CXM-03, CXM-09, CXM-12, CXM-15, CXM-18, CXM-20, CXM-24 Both the 2 μΜ and 0.2 μΜ concentrations showed better or comparable activity than the positive control apatinib.
实施例 35 测试本发明化合物对不同肿瘤细胞株的 IC5Q值 Example 35 Testing IC 5Q values of compounds of the invention against different tumor cell lines
1、 实验材料 1. Experimental materials
本发明的化合物 CXM-30、 33、 02、 05、 12、 09、 24；阳性对照 BIBW-2992, 拉帕替尼： 购自杭州荣大医药化工有限公司； 非小细胞癌细胞 NCI-H322M、 前列腺癌细胞 DU145: 购自美国菌种保藏中心 ATCC; 肝癌细胞 Hep3B、 乳 腺癌细胞 MDA-MB-231、 卵巢癌细胞 SK-OV-3、 下咽癌人咽鳞癌细胞 FaDu、 恶性胶质母细胞瘤细胞 U-87MG: 购自中科院上海细胞生物研究所； CCK-8 检测试剂盒： 购自 Cat# CK04-13， Dojindo公司。  The compound of the invention CXM-30, 33, 02, 05, 12, 09, 24; positive control BIBW-2992, lapatinib: purchased from Hangzhou Rongda Pharmaceutical Chemical Co., Ltd.; non-small cell cancer cell NCI-H322M, Prostate cancer cell line DU145: purchased from American Type Culture Collection ATCC; liver cancer cell Hep3B, breast cancer cell MDA-MB-231, ovarian cancer cell SK-OV-3, hypopharyngeal carcinoma human pharyngeal carcinoma cell FaDu, malignant collagen Cell tumor cell U-87MG: purchased from Shanghai Institute of Cell Biology, Chinese Academy of Sciences; CCK-8 test kit: purchased from Cat# CK04-13, Dojindo.
2、 实验方法： 2. Experimental method:
1 ) 细胞培养  1) Cell culture
各细胞株用以下相应的完全培养基培养， 细胞置于在 37 °C， 100 %相对 湿度， 5 % 02培养箱中生长， 选择对数生长期的细胞进行实验测试。 Each cell line was cultured with the corresponding complete medium below, and the cells were grown in a incubator at 37 ° C, 100% relative humidity, 5 % 0 2 , and cells in logarithmic growth phase were selected for experimental testing.
培养基的配制如下表 7所示。  The preparation of the medium is shown in Table 7 below.
表 7  Table 7
Figure imgf000036_0001
Figure imgf000036_0001
a) 收集对数生长期细胞， 计数， 用完全培养基重新悬浮细胞；  a) collect logarithmic growth phase cells, count, and resuspend the cells with complete medium;
b)调整细胞浓度至合适浓度， 接种 96孔板， 每孔接种 100 μΐ细胞悬液; c) 细胞在 37 °C， 100 %相对湿度， 5 % C02培养箱中孵育 24h。 b) Adjust the cell concentration to the appropriate concentration, inoculate 96-well plates, inoculate 100 μΐ cell suspension per well; c) Incubate the cells for 24 h at 37 °C, 100% relative humidity, 5 % C0 2 incubator.
2) IC5。实验 a) 收集对数生长期细胞， 计数， 用完全培养基重新悬浮细胞， 调整细胞 浓度至合适浓度 （依照细胞密度优化试验结果确定）， 接种 96孔板， 每孔加 ΙΟΟμΙ细胞悬液。 细胞在 37°C， 100 %相对湿度， 5% C02培养箱中孵育 24h。 2) IC 5 . experiment a) Collect logarithmic growth phase cells, count, resuspend the cells with complete medium, adjust the cell concentration to the appropriate concentration (determined according to the cell density optimization test results), inoculate 96-well plates, and add ΙΟΟμΙ cell suspension to each well. The cells were incubated for 24 h at 37 ° C, 100% relative humidity, 5% C0 2 incubator.
b)用培养基将待测化合物稀释至 5μΜ &0.5μΜ， 按 25μ1/孔加入细胞， 化 合物终浓度为 1 μΜ至 0.1 μΜ。  b) Dilute the test compound to 5 μΜ & 0.5 μΜ with medium, and add the cells to 25 μl/well to a final concentration of 1 μΜ to 0.1 μΜ.
c) 细胞置于 37 °C， 100 %相对湿度， 5% C02培养箱中孵育 72h。 c) The cells were incubated for 72 h at 37 ° C, 100% relative humidity, 5% C0 2 incubator.
d)吸弃培养基，加入含 10% CCK-8的完全培养基置于 37 °C培养箱中孵 育 2-4h。  d) Aspirate the medium and add to the complete medium containing 10% CCK-8 and incubate in a 37 °C incubator for 2-4 h.
e) 轻轻震荡后在 SpectraMax M5 Microplate Reader上测定 450 nm波长 处的吸光度， 以 650 nm 处吸光度 (OAM乍为参比， 计算抑制率。  e) The absorbance at 450 nm was measured on a SpectraMax M5 Microplate Reader with gentle shaking, and the absorbance at 650 nm (OAM乍 was used as a reference to calculate the inhibition rate.
3 ) 数据计算  3) Data calculation
肿瘤细胞生长抑制率％ = [( 0八8)/( (；- )：^100%  Tumor cell growth inhibition rate % = [( 0 8 8) / ( (;- ): ^ 100%
As: 样品的吸光度 (细胞 +CCK-8+待测化合物）  As: absorbance of the sample (cell + CCK-8 + test compound)
Ac: 阴性对照的吸光度 （细胞 +CCK-8+DMSO)  Ac: Absorbance of the negative control (cell + CCK-8 + DMSO)
Ab: 阳性对照的吸光度 （培养基 +CCK-8+DMSO)  Ab: absorbance of positive control (medium + CCK-8 + DMSO)
按照说明书的方法， 用 CCK-8检测试剂盒检测本发明的化合物及阳性对 照分别对不同细胞株 （非小细胞癌细胞 NCI-H322M、 肝癌细胞 Hep3B、 乳 腺癌细胞 MDA-MB-231、前列腺癌细胞 DU145、卵巢癌细胞 SK-OV-3、下咽 癌人咽鳞癌细胞 FaDu、恶性胶质母细胞瘤细胞 U-87MG )的体外细胞毒性 IC5。 值。用 DMSO稀释化合物作用终浓度为从 ΙΟμΜ至 0 μΜ， 包括 10μΜ、 2μΜ、 0·4μΜ、 0·08μΜ、 0·016μΜ、 0·0032μΜ、 0·00064μΜ、 0.000128μΜ和 0μΜ。 3、 结果与分析 According to the method of the specification, the compound of the present invention and the positive control were separately detected by the CCK-8 test kit for different cell lines (non-small cell cancer cell NCI-H322M, liver cancer cell Hep3B, breast cancer cell MDA-MB-231, prostate cancer). In vitro cytotoxicity IC 5 of cell DU145, ovarian cancer cell SK-OV-3, hypopharyngeal carcinoma human pharyngeal carcinoma cell line FaDu, glioblastoma cell line U-87MG. value. The final concentration of the compound diluted with DMSO was from ΙΟμΜ to 0 μΜ, including 10 μΜ, 2 μΜ, 0·4 μΜ, 0·08 μΜ, 0·016 μΜ, 0·0032 μΜ, 0·00064 μΜ, 0.000128 μΜ, and 0 μΜ. 3. Results and analysis
实验结果如表 7所示。  The experimental results are shown in Table 7.
表 7 化合物对不同肿瘤细胞株的体外细胞毒性 IC5Q值 Table 7 In vitro cytotoxicity IC 5Q values of compounds against different tumor cell lines
Figure imgf000037_0001
Hep3B 0.5328 0.6676 0.2798 0.7636 0.2004 0.2438 1.083 1.014 2.728
Figure imgf000037_0001
Hep3B 0.5328 0.6676 0.2798 0.7636 0.2004 0.2438 1.083 1.014 2.728
Fadu 0.2169 0.1166 0.1176 0.1969 0.04038 0.02517 0.0227 0.06814 0.5656 从表 7可以看出， 化合物 CXM-02 CXM-12 CXM-24表现出比阳性对 照药 BIBW-2992、 拉帕替尼较高的生物活性； 化合物 CXM-30 CXM-09 CXM-33 表现出比阳性对照 BIBW-2992、 拉帕替尼相当的生物活性； 化合物 CXM-05表现出比阳性对照 BIBW-2992、 拉帕替尼略低的生物活性。 Fadu 0.2169 0.1166 0.1176 0.1969 0.04038 0.02517 0.0227 0.06814 0.5656 As can be seen from Table 7, the compound CXM-02 CXM-12 CXM-24 showed higher biological activity than the positive control drug BIBW-2992, lapatinib; Compound CXM- 30 CXM-09 CXM-33 exhibited a biological activity comparable to the positive control BIBW-2992, lapatinib; Compound CXM-05 exhibited a lower biological activity than the positive control BIBW-2992, lapatinib.
实施例 36 本发明化合物在大鼠体内的药代动力学行为研究 Example 36 Pharmacokinetic Behavior of the Compounds of the Invention in Rats
1、 实验材料：  1. Experimental materials:
动物： 清洁级 SD大鼠 96只， 雌性， 体重 >200克；  Animals: 96 SD rats, female, weighing >200 g;
试剂：本发明的化合物： CXM-02 CXM-30 CXM-33 CXM- 12 CXM-09 CXM-24; 阳性对照品： 拉帕替尼 （杭州荣大医药化工有限公司产品）、 BIBW -2992 (杭州荣大医药化工有限公司产品公司产品）；  Reagents: Compounds of the invention: CXM-02 CXM-30 CXM-33 CXM- 12 CXM-09 CXM-24; Positive control: Lapatinib (product of Hangzhou Rongda Pharmaceutical Chemical Co., Ltd.), BIBW -2992 (Hangzhou Rongda Pharmaceutical Chemical Co., Ltd. Product Company);
仪器： 液相色谱仪 （安捷伦公司 AgilentllOO); 质谱仪 （API4000, 美国 应用生物系统公司）， 电喷雾离子源 （ESI), 串联四极杆质量分析器。 数据处 理系统为 Analyst软件 （美国应用生物系统公司， 软件版本号 1.5.1 )。  Instruments: Liquid Chromatograph (AgilentllOO); Mass Spectrometer (API4000, Applied Biosystems, USA), Electrospray Ionization (ESI), Tandem Quadrupole Mass Analyzer. The data processing system is Analyst Software (Applied Biosystems, USA, software version number 1.5.1).
2、 实验方法：  2. Experimental method:
将大鼠 48只， 随机分为 8组， 每组 6只， 分别按照 2mg/kg体重经单次 静脉注射施予本发明的 6个化合物和 2个阳性对照样品。同时，将大鼠 48只， 随机分为 8组， 每组 6只， 分别按照 5mg/kg体重经单次口服施予本发明的 6 个化合物和 2个阳性对照品。在给药前， 给药后 5 min 15 min 30 min 1 h 2 h 4 h 6 h 8 h和 24h， 每只动物分别经眼眶静脉 （异氟垸麻醉后） 采血 约 0.2 mL， 肝素钠抗凝。 血样置于冰上， 并于 30min之内离心分离血桨 （离 心条件： 8000r/min 6min 4°C)。 收集的血桨分析前存放于 -80°C  Forty-eight rats were randomly divided into 8 groups, 6 in each group, and 6 compounds and 2 positive control samples of the present invention were administered by single intravenous injection at 2 mg/kg body weight. At the same time, 48 rats were randomly divided into 8 groups, 6 in each group, and 6 compounds of the present invention and 2 positive controls were orally administered in a single oral dose of 5 mg/kg body weight. Before administration, 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, 8 h, and 24 h after administration, each animal was subjected to blood collection of about 0.2 mL via the orbital vein (after anesthesia with isoflurane). Heparin sodium anticoagulation . The blood sample was placed on ice and the blood paddle was centrifuged within 30 min (centrifugal condition: 8000 r/min 6 min 4 ° C). Store the collected blood plasma before storage at -80 ° C
采用液相色谱 -质谱 -质谱 （LC/MS/MS )联用法测定大鼠血桨中受试物的 浓度。根据所得的血药浓度数据采用药动学处理软件 WinNonlin 5.2非房室模 型计算相关药代动力学参数。  The concentration of the test substance in the rat blood plasma was determined by liquid chromatography-mass spectrometry-mass spectrometry (LC/MS/MS). Based on the obtained blood drug concentration data, the pharmacokinetic processing software WinNonlin 5.2 non-compartment model was used to calculate the relevant pharmacokinetic parameters.
3、 结果与分析 3. Results and analysis
实验结果如下表 8和表 9所示：  The experimental results are shown in Table 8 and Table 9 below:
表 8 大鼠单次静脉给予本发明的化合物及 2个阳性对照品的主要药动学参数  Table 8 Main pharmacokinetic parameters of a single intravenous administration of a compound of the invention and two positive controls in rats
Figure imgf000038_0001
33.33 39.55 22.69 25.22 7.81 54.84 52.06 ±291.56
Figure imgf000038_0001
33.33 39.55 22.69 25.22 7.81 54.84 52.06 ±291.56
1.87±0. 16.47士 5.48士 4.47士 8.99士 3.78士 3.45士 1.34士 ti/2z (hr) 1.87±0. 16.47 士 5.48士 4.47士 8.99士 3.78士 3.45士 1.34士 ti/2z (hr)
16 12.03 0.95 3.03 9.95 1.88 1.45 0.48 16 12.03 0.95 3.03 9.95 1.88 1.45 0.48
20.10士 23.07士 39.44士 35.81士 68.50士 22.48士 14.09士 1.90士20.10 士 23.07士 39.44士 35.81士 68.50士 22.48士 14.09士 1.90士
Vz(L/kg) V z (L/kg)
6.19 4.36 7.29 12.40 34.99 8.07 4.05 0.66 6.19 4.36 7.29 12.40 34.99 8.07 4.05 0.66
7.41士 1.24士 5.04士 6.35士 8.15士 4.34士 2.94士 0.99士7.41 士 1.24士 5.04士 6.35士 8.15士 4.34士 2.94士 0.99士
Clz(L/hr/kg) Cl z (L/hr/kg)
1.84 0.56 1.00 1.79 4.05 0.68 0.39 0.02 1.84 0.56 1.00 1.79 4.05 0.68 0.39 0.02
1206.49 2013.341206.49 2013.34
255.66士 251.21士 301.60士 147.35士 423.33士 635.33士 255.66 士 251.21 301.60 士 147.35 423.33 635.33 士
AUC ( g-hr/L) 士 士  AUC ( g-hr/L)
57.04 30.99 102.04 22.29 86.24 114.00  57.04 30.99 102.04 22.29 86.24 114.00
256.64 37.97 表 9 大鼠单次口服给予本发明的化合物及阳性对照品的主要药动学参数  256.64 37.97 Table 9 Main pharmacokinetic parameters of a single oral administration of a compound of the invention and a positive control in rats
Figure imgf000039_0001
Figure imgf000039_0001
从表 8和表 9的结果可以看出， 化合物 CXM-33、 CXM-02、 CXM-24表 现出比阳性对照药 BIBW-2992、 拉帕替尼较好的口服生物利用度； 化合物 CXM-09、 CXM-30, CXM-12表现出与阳性对照药 BIBW-2992、 拉帕替尼相 近或略低的口服生物利用度。 6个化合物单次静脉的主要药代动力学参数超过 了仪器测量的上限， 因此静脉给药的方式不适用该类化合物的制剂方式。  From the results of Tables 8 and 9, it can be seen that the compounds CXM-33, CXM-02, CXM-24 showed better oral bioavailability than the positive control drug BIBW-2992, lapatinib; Compound CXM-09 CXM-30, CXM-12 showed similar or slightly lower oral bioavailability than the positive control drug BIBW-2992, lapatinib. The primary pharmacokinetic parameters of the single vein of the six compounds exceeded the upper limit of the instrumental measurement, so the manner of intravenous administration did not apply to the formulation of such compounds.

Claims

权利要求书  Claim
Figure imgf000040_0001
Figure imgf000040_0001
其中， 和 独立地表示氢、 卤素、 CM垸基、 卤素取代的 CM垸基、Wherein, and independently represent hydrogen, halogen, C M fluorenyl, halogen-substituted CM thiol,
CM垸氧基、 卤素取代的 d_4垸氧基、 腈基、 酯基、 硝基、 氨基、 酰胺基、 磺 酰胺基、 苯基或 CH3S02CH2CH2NHCH2-Ar; 其中 Ar表示苯基、 呋喃基、 噻吩 基、 吡咯基和噻唑基， 并且 Ar可被一个或两个选自卤素、 d_4垸基和 CM垸 氧基的基团取代； 或者 C M methoxy, halogen-substituted d 4 methoxy, cyano, ester, nitro, amino, amide, sulfonamide, phenyl or CH 3 S0 2 CH 2 CH 2 NHCH 2 -Ar ; Ar represents a phenyl group, a furyl group, a thienyl group, a pyrrolyl group and a thiazolyl group, and Ar may be substituted by one or two groups selected from the group consisting of halogen, d- 4 fluorenyl and CM methoxy; or
R R2独立地表示： RR 2 stands independently:
X-(CH2)n-R4 X-(CH 2 ) n -R 4
其中 X表示 N、 0或 S; n表示 0到 6的整数； R4表示氢、 CM垸基、 卤 素取代的 (^_4垸基、 （^_4垸氧基、 卤素取代的 (^_4垸氧基、 呋喃基、 四氢呋喃 基、 吗啉基、 吡喃基、 四氢吡喃基、 吡咯基、 吡啶基、 噁唑啉基、 噻唑基或噻 吩基； 或者 Wherein X represents N, 0 or S; n represents an integer from 0 to 6; R 4 represents hydrogen, C M fluorenyl, halogen substituted (^_ 4 fluorenyl, (^_ 4 fluorenyl, halogen substituted (^) _ 4 methoxy, furyl, tetrahydrofuranyl, morpholinyl, pyranyl, tetrahydropyranyl, pyrrolyl, pyridyl, oxazolinyl, thiazolyl or thienyl;
Ri, R2独立地表示：
Figure imgf000040_0002
Ri, R 2 stands independently:
Figure imgf000040_0002
其中 R5表示 Η、 Ν, Ν-二甲基胺基甲基、 Ν, Ν-二乙基胺基甲基或 Ν, Ν-二 丙基胺基甲基； Wherein R 5 represents hydrazine, hydrazine, hydrazine-dimethylaminomethyl, hydrazine, hydrazine-diethylaminomethyl or hydrazine, fluorenyl-dipropylaminomethyl;
R3表示氢或 C1-4院基。 R 3 represents hydrogen or a C 1-4 yard group.
2、 如权利要求 1所述的化合物， 其中，  2. The compound according to claim 1, wherein
和 独立地表示氢、 CM垸氧基、 硝基、 氨基、 酰胺基； 或者 And independently represent hydrogen, C M methoxy, nitro, amino, amide; or
Ri, R2独立地表示： Ri, R 2 stands independently:
X-(CH2)n-R4 X-(CH 2 ) n -R 4
其中 X表示 0或 S; n表示 0到 4的整数； 表示氢、 CM垸基、 CM垸 氧基、 3-四氢呋喃基、 吗啉基、 4-四氢吡喃基、 2-吡咯基、 噁唑啉基、 噻唑基 或噻吩基； 或者， Ri, R2独立地表示：
Figure imgf000041_0001
Wherein X represents 0 or S; n represents an integer from 0 to 4; represents hydrogen, CM thiol, C M methoxy, 3-tetrahydrofuranyl, morpholinyl, 4-tetrahydropyranyl, 2-pyrrolyl, Oxazolinyl, thiazolyl or thienyl; or Ri, R 2 stands independently:
Figure imgf000041_0001
其中 R5表示 H、 N, N-二甲基胺基甲基、 Ν, Ν-二乙基胺基甲基或 Ν, Ν-二 丙基胺基甲基； Wherein R 5 represents H, N, N-dimethylaminomethyl, hydrazine, hydrazine-diethylaminomethyl or hydrazine, fluorenyl-dipropylaminomethyl;
R3表示氢。 R 3 represents hydrogen.
3、 如权利要求 1所述的化合物， 其中所述化合物选自：  3. A compound according to claim 1 wherein the compound is selected from the group consisting of:
N-{4-[l-(3-氟-苄基） 吲唑 -5-基胺基] -喹唑啉 -6-基 丙烯酰胺 (CXM-01);  N-{4-[l-(3-Fluoro-benzyl)carbazole-5-ylamino]-quinazoline-6-yl acrylamide (CXM-01);
( - {4-[1-(3-氟-苄基) 吲唑 -5-基胺基 ]-7-甲氧基-喹唑啉 -6-基}-4- (二 甲基氨基)丁 -2-烯酰胺 （CXM-02);  (-{4-[1-(3-Fluoro-benzyl)oxazol-5-ylamino]-7-methoxy-quinazolin-6-yl}-4-(dimethylamino) butyl -2-enamide (CXM-02);
( - {4-[1-(3-氟-苄基) 吲唑 -5-基胺基 ]-7-乙氧基-喹唑啉 -6-基}-4- (二 甲基胺基) -丁 -2-烯酰胺 （CXM-03);  (-{4-[1-(3-Fluoro-benzyl)oxazol-5-ylamino]-7-ethoxy-quinazolin-6-yl}-4-(dimethylamino) -but-2-enamide (CXM-03);
7-(2-甲氧基乙氧基) -N-[l-(3-氟-苄基) 吲唑 -5-基] -6-硝基-喹唑啉 -4-胺 (CXM-04);  7-(2-methoxyethoxy)-N-[l-(3-fluoro-benzyl)oxazol-5-yl]-6-nitro-quinazolin-4-amine (CXM-04 );
( -Ν-{4-[1-(3-氟-苄基) 吲唑 -5-基胺基 ]-7-(3-甲氧基乙氧基) -喹唑啉 -6- 基}-4- (二甲基胺基) -丁 -2-烯酰胺 （CXM-05); (- Ν - {4- [1- (3- fluoro-benzyl) - amino-indazol-5-yl] -7- (3-methoxy-ethoxy) - quinazolin-6-yl} - 4-(Dimethylamino)-but-2-enamide (CXM-05);
N-{4-[l-(3-氟-苄基) 吲唑 -5-基胺基 ]-7-(2-甲氧乙氧基) -喹唑啉 -6-基 丙烯酰胺 （CXM-06);  N-{4-[1-(3-Fluoro-benzyl)oxazol-5-ylamino]-7-(2-methoxyethoxy)-quinazolin-6-ylacrylamide (CXM- 06);
N-{4-[l-(3-氟-苄基) 吲唑 -5-基胺基 ]-7-[3-(4-吗啉代)丙氧基] -喹唑啉 -6- 基} -丙烯酰胺 （CXM-07 );  N-{4-[1-(3-Fluoro-benzyl)oxazol-5-ylamino]-7-[3-(4-morpholino)propoxy]-quinazoline-6-yl } - acrylamide (CXM-07);
( - {4-[1-(3-氟-苄基) -1H-吲唑 -5-基胺基 ]-7-[3-(4-吗啉代)丙氧基] -喹唑 啉 -6-基}-4- (二甲基胺基) -丁 -2-烯酰胺 （CXM-08); ( -{4-[1-(3-Fluoro-benzyl)-1H-indazol-5-ylamino]-7-[3-(4-morpholino)propoxy]-quinazoline- 6-yl}-4-(dimethylamino)-but-2-enamide (CXM-08);
( )-N-{7-[0¾-四氢呋喃 -3-基氧基 ]-4-[1-(3-氟-苄基) 吲唑 -5-基胺基] -喹 唑啉 -6-基}-4-二甲基胺基-丁 -2-烯酰胺 （CXM-09);  ( )-N-{7-[03⁄4-tetrahydrofuran-3-yloxy]-4-[1-(3-fluoro-benzyl)oxazol-5-ylamino]-quinazolin-6-yl }-4-dimethylamino-but-2-enamide (CXM-09);
( )-W-{4-[l-(3-氟-苄基) 吲唑 -5-基胺基 ]-7- (腈基) -喹唑啉 -6-基}-4-二甲 基胺基-丁 -2-烯酰胺 （CXM-10);  ( )-W-{4-[l-(3-Fluoro-benzyl)oxazol-5-ylamino]-7-(nitrile)-quinazolin-6-yl}-4-dimethyl Amino-but-2-enamide (CXM-10);
( -N-{7- [(四氢 -2H-吡喃 -4-基) -甲硫基] -4-[1-(3-氟-苄基) 吲唑 -5-基胺 基] -喹唑啉 -6-基}-4-二甲基胺基-丁 -2-烯酰胺 （CXM-11);(-N-{7-[(tetrahydro-2H-pyran-4-yl)-methylthio]-4-[1-(3-fluoro-benzyl)oxazol-5-ylamino] - Quinazoline- 6 -yl}-4-dimethylamino-but-2-enamide (CXM-11);
- {4-[1-(3-氟-苄基) 吲唑 -5-基胺基] -喹唑啉 -6-基}-4-二甲基胺基- 丁 -2-烯酰胺 （CXM-12);  - {4-[1-(3-Fluoro-benzyl)oxazol-5-ylamino]-quinazolin-6-yl}-4-dimethylamino-but-2-enamide (CXM -12);
N-{4-[l-(3-氟 -苄基 )-1^-吲唑 -5-基胺基 ]-7-甲氧基-喹唑啉 -6-基 丙烯酰胺 (CXM-13);  N-{4-[1-(3-Fluoro-benzyl)-1^-oxazol-5-ylamino]-7-methoxy-quinazolin-6-yl acrylamide (CXM-13) ;
N-{4-[l-(3-氟-苄基) 吲唑 -5-基胺基 ]-7-乙氧基-喹唑啉 -6-基 丙烯酰胺 (CXM-14); N-{7-[( -四氢呋喃 -3-基氧基 ]-4-[l-(3-氟-苄基) 吲唑 -5-基胺基] -喹唑啉 -6-基} -丙烯酰胺 （CXM-15); N-{4-[l-(3-Fluoro-benzyl)oxazol-5-ylamino]-7-ethoxy-quinazolin-6-yl acrylamide (CXM-14); N-{7-[(-tetrahydrofuran-3-yloxy]-4-[l-(3-fluoro-benzyl)oxazol-5-ylamino]-quinazolin-6-yl}-propene Amide (CXM-15);
N-{4-[l-(3-氟-苄基) 吲唑 -5-基胺基 ]-7-腈基-喹唑啉 -6-基 丙烯酰胺 (CXM-16);  N-{4-[l-(3-fluoro-benzyl)carbazole-5-ylamino]-7-cyano-quinazoline-6-yl acrylamide (CXM-16);
N-{7- [(四氢 -2H-吡喃 -4-基) -甲硫基] -4-[1-(3-氟-苄基) 吲唑 -5-基胺基] - 喹唑啉 -6-基 丙烯酰胺 (CXM-17);  N-{7-[(tetrahydro-2H-pyran-4-yl)-methylthio]-4-[1-(3-fluoro-benzyl)oxazol-5-ylamino]-quinazoline Porphyrin-6-yl acrylamide (CXM-17);
N-{4-[l-(3-氟-苄基） 吲唑 -5-基胺基] -喹唑啉 -7-基 丙烯酰胺 (CXM-18);  N-{4-[l-(3-fluoro-benzyl)carbazole-5-ylamino]-quinazoline-7-yl acrylamide (CXM-18);
( - {4-[1-(3-氟-苄基) 吲唑 -5-基胺基 ]-6-甲氧基-喹唑啉 -7-基}-4-二 甲基胺基-丁 -2-烯酰胺 （CXM-19);  (-{4-[1-(3-Fluoro-benzyl)oxazol-5-ylamino]-6-methoxy-quinazolin-7-yl}-4-dimethylamino-butyl -2-enamide (CXM-19);
( - {4-[1-(3-氟-苄基) 吲唑 -5-基胺基 ]-6-乙氧基-喹唑啉 -7-基}-4-二 甲基胺基-丁 -2-烯酰胺 （CXM-20);  (-{4-[1-(3-Fluoro-benzyl)oxazol-5-ylamino]-6-ethoxy-quinazolin-7-yl}-4-dimethylamino-butyl -2-enamide (CXM-20);
( -^-{4-[1-(3-氟-苄基) 吲唑 -5-基胺基 ]-6-(2-甲氧基乙氧基) -喹唑啉 -7- 基}-4-二甲基胺基-丁 -2-烯酰胺 （CXM-21); (-^-{4-[1-(3-Fluoro-benzyl)oxazol-5-ylamino]-6-(2-methoxyethoxy)-quinazolin-7-yl}- 4-dimethylamino-but-2-enamide (CXM-21);
{4-[1-(3-氟-苄基) 吲唑 -5-基胺基 ]-6-(2-甲氧基乙氧基) -喹唑啉 -7- 基 丙烯酰胺 （CXM-22);  {4-[1-(3-Fluoro-benzyl)oxazol-5-ylamino]-6-(2-methoxyethoxy)-quinazolin-7-yl acrylamide (CXM-22 );
N-{4-[l-(3-氟-苄基) 吲唑 -5-基胺基 ]-6-[3-(4-吗啉代)丙氧基] -喹唑啉 -7- 基 丙烯酰胺 （CXM-23);  N-{4-[l-(3-Fluoro-benzyl)oxazol-5-ylamino]-6-[3-(4-morpholino)propoxy]-quinazolin-7-yl Acrylamide (CXM-23);
( - {4-[1-(3-氟-苄基) 吲唑 -5-基胺基 ]-6-[3-(4-吗啉代)丙氧基] -喹唑 啉 -7-基}-4-二甲基胺基-丁 -2-烯酰胺 （CXM-24); ( -{4-[1-(3-Fluoro-benzyl)oxazol-5-ylamino]-6-[3-(4-morpholino)propoxy]-quinazolin-7-yl }-4-dimethylamino-but-2-enamide (CXM-24);
( )-N-{6-[0¾-四氢呋喃 -3-基氧基 ]-4-[1-(3-氟-苄基) 吲唑 -5-基胺基] -喹 唑啉 -7-基}-4-二甲基胺基-丁 -2-烯酰胺 （CXM-25);  ( )-N-{6-[03⁄4-tetrahydrofuran-3-yloxy]-4-[1-(3-fluoro-benzyl)oxazol-5-ylamino]-quinazolin-7-yl }-4-dimethylamino-but-2-enamide (CXM-25);
( - {4-[1-(3-氟-苄基) 吲唑 -5-基胺基 ]-6-腈基-喹唑啉 -7-基}-4-二甲 基胺基-丁 -2-烯酰胺 （CXM-26); ( -{4-[1-(3-Fluoro-benzyl)oxazol-5-ylamino]-6-cyano-quinazolin-7-yl}-4-dimethylamino-butyl- 2-enamide (CXM-26);
( )-W-{6- [(四氢 -2H-吡喃 -4-基) -甲硫基] -4-[1-(3-氟-苄基) 吲唑 -5-基胺 基] -喹唑啉 -7-基}-4-二甲基胺基-丁 -2-烯酰胺 （CXM-27);( )-W-{6-[(tetrahydro-2H-pyran-4-yl)-methylthio]-4-[1-(3-fluoro-benzyl)oxazol-5-ylamino] -quinazoline- 7 -yl}-4-dimethylamino-but-2-enamide (CXM-27);
- {4-[1-(3-氟-苄基) 吲唑 -5-基胺基] -喹唑啉 -7-基}-4-二甲基胺基- 丁 -2-烯酰胺 （CXM-28);  - {4-[1-(3-Fluoro-benzyl)oxazol-5-ylamino]-quinazolin-7-yl}-4-dimethylamino-but-2-enamide (CXM -28);
N-{4-[l-(3-氟 -苄基 )-1^-吲唑 -5-基胺基 ]-6-甲氧基-喹唑啉 -7-基 丙烯酰胺 (CXM-29);  N-{4-[1-(3-Fluoro-benzyl)-1^-indazol-5-ylamino]-6-methoxy-quinazolin-7-yl acrylamide (CXM-29) ;
N-{4-[l-(3-氟-苄基) 吲唑 -5-基胺基 ]-6-乙氧基-喹唑啉 -7-基 丙烯酰胺 (CXM-30);  N-{4-[1-(3-Fluoro-benzyl)carbazole-5-ylamino]-6-ethoxy-quinazolin-7-yl acrylamide (CXM-30);
N-{6-[0¾-四氢呋喃 -3-基氧基 ]-4-[1-(3-氟-苄基) 吲唑 -5-基胺基] -喹唑啉 -7-基} -丙烯酰胺 （CXM-31); N-{6-[03⁄4-tetrahydrofuran-3-yloxy]-4-[1-(3-fluoro-benzyl)oxazol-5-ylamino]-quinazolin-7-yl}-propene Amide (CXM-31);
{4-[1-(3-氟-苄基) 吲唑 -5-基胺基 ]-6-腈基-喹唑啉 -7-基 丙烯酰胺 (CXM-32); {4-[1-(3-Fluoro-benzyl)oxazol-5-ylamino]-6-cyano-quinazoline-7-yl acrylamide (CXM-32);
N-{6- [(四氢 -2H-吡喃 -4-基) -甲硫基] -4-[l-(3-氟-苄基) 吲唑 -5-基胺基] - 喹唑啉 -7-基} -丙烯酰胺 （CXM-33)。  N-{6-[(tetrahydro-2H-pyran-4-yl)-methylthio]-4-[l-(3-fluoro-benzyl)oxazol-5-ylamino]-quinazoline Porphyrin-7-yl}-acrylamide (CXM-33).
4、 如权利要求 1所述的化合物， 其中所述化合物为互变异构体、 立体异构体 或其药学上可接受的盐的形式。  4. A compound according to claim 1 wherein the compound is in the form of a tautomer, a stereoisomer or a pharmaceutically acceptable salt thereof.
5、 如权利要求 4所述的化合物， 其中所述药学上可接受的盐为所述化合物与 酸形成的酸加成盐， 所述酸选自氢溴酸、 盐酸、 硫酸、 亚硫酸、 乙酸、 草酸、 戊酸、 油酸、 棕榈酸、 硬脂酸、 月桂酸、 硼酸、 苯甲酸、 乳酸、 磷酸、 甲苯甲 酸、 柠檬酸、 马来酸、 富马酸、 琥珀酸、 酒石酸、 苯甲酸、 甲磺酸、 葡萄糖酸、 乳糖酸和月桂基磺酸。  The compound according to claim 4, wherein the pharmaceutically acceptable salt is an acid addition salt of the compound with an acid selected from the group consisting of hydrobromic acid, hydrochloric acid, sulfuric acid, sulfurous acid, acetic acid. , oxalic acid, valeric acid, oleic acid, palmitic acid, stearic acid, lauric acid, boric acid, benzoic acid, lactic acid, phosphoric acid, toluic acid, citric acid, maleic acid, fumaric acid, succinic acid, tartaric acid, benzoic acid, Methanesulfonic acid, gluconic acid, lactobionic acid and lauryl sulfonic acid.
6、 一种药物组合物， 包含权利要求 1至 4任一项所述的化合物以及药学上可 接受的载体。  A pharmaceutical composition comprising the compound according to any one of claims 1 to 4 and a pharmaceutically acceptable carrier.
7、 权利要求 1至 4任一项所述的化合物在制备药物中的用途， 所述药物用于 治疗由蛋白络氨酸激酶介导的疾病。  7. Use of a compound according to any one of claims 1 to 4 for the manufacture of a medicament for the treatment of a disease mediated by a protein tyrosine kinase.
8、 权利要求 7所述的用途， 其中所述疾病为乳腺癌、 非小细胞肺癌、 卵巢癌、 胃癌或胰腺癌。  8. The use of claim 7, wherein the disease is breast cancer, non-small cell lung cancer, ovarian cancer, gastric cancer or pancreatic cancer.
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