Indolone derivatives that are replaced by pyrrole, the method of preparation thereafter, the components that make up the same substance and their uses.

Abstract

OCR04PP The present invention involves an indolone derivative which is replaced by pyrrole, a method. The preparation thereto, the components comprising derivatives, and the applications of those. The indolone derivative which is replaced by -pyrrol has a structure which is shown in formula(I) below. The current invention also involves the use of indolone derivatives which are replaced by... Pyrrol is used for therapeutic purposes through tyrosine kinase receptor-mediated therapy, and its components are linked to other treatments. Pharmaceuticals containing compounds with that structure for the treatment of related diseases, such as: tumor (Chemical formula)(I)

Description

The present invention relates to a pyrrole-substituted indolinone derivative or a pharmaceutically acceptable salt thereof, a preparation method thereof, a composition containing the derivative, and an application thereof. More specifically, the present invention relates to a multi-target phenolic compound An amino acid kinase inhibitor pyrrole-substituted indolinone derivative, as well as its pharmaceutical composition and medical application. Cancer is the disease that threatens people's health the most in today's society. Anticancer drugs widely used in the market so far are still some cytotoxic drugs discovered in the last century. These cytotoxic drugs kill a large number of normal cells in the process of treating tumors and bring unbearable side effects to patients, and with the extensive use of such drugs, drug resistance is becoming another difficult problem to overcome. Tumor blood vessel inhibition is a new method developed at the end of last century for the treatment of tumors. Its research basis is based on Folkman's viewpoint that tumor survival, growth and metastasis depend on a large number of new blood vessels (Folkman.J.et.al.N.Engl.J.Med., 1971, 285, 1182-1186.). A large number of clinical findings have found that tumor tissue contains a large number of new blood vessels, and the growth and metastasis of tumor cells require a large number of blood vessels to provide sufficient oxygen and nutrients. Inhibiting the growth of tumor cell blood vessels can "starve" tumor cells to death. At the same time, there are few new blood vessels around normal cells, and the inhibition of new blood vessels has little effect on normal cells. Toxic and other characteristics. Vascular inhibition can be divided into direct inhibition and indirect inhibition. Direct inhibition acts on endothelial cells of blood vessels to inhibit angiogenesis, expansion and nutritional support for tumor cells. The main method currently used is rhythmicization of cytotoxic drugs. Treatment, although rhythmic treatment can reduce the side effects of cytotoxic drugs, it is still difficult to change the damage of drugs to the human body. Indirect inhibition inhibits angiogenesis by inhibiting angiogenesis factors required in the process of angiogenesis (Cao, Y. et. al. Int. J. Biochem. Cell Biol., 2001, 33, 357-369.). The process of angiogenesis includes: vascular endothelial cells are activated under the action of activating factors; endothelial cells secrete proteases to degrade the basement membrane; endothelial cell migration and proliferation; formation of new capillary lumens; recruitment of pericytes to stabilize the newly generated capillaries peripheral structure. Under physiological conditions, there are two types of factors that act on angiogenesis, namely, angiogenesis inhibitory factors and pro-angiogenic factors. Angiogenesis inhibitors can be divided into two categories according to the specificity of their actions. One is angiogenesis inhibitory factors that specifically act on endothelial cells, including angiostatin and endostatin. The other is angiogenesis inhibitory factors that act non-specifically on endothelial cells, including cytokines, tissue metalloproteinase inhibitors, serine protease inhibitors, and tumor suppressor gene products. Pro-angiogenic factors include epidermal growth factor (EGF), endothelial growth factor (VEGF), platelet-derived growth factor (PDGF) and fibroblast growth factor (FGF) etc. (Hanks, S.K., et.al.FASEB, 1995, 9, 576-696). The high expression of different pro-angiogenic factors can be seen in different types of tumors, such as the high expression of EGF in epithelial cell tumors and the high expression of PDGF in glioma. The current strategies for developing anti-cancer drugs targeting tumor neovascularization pathways are mainly to increase angiogenesis inhibitors and reduce pro-angiogenic factors, among which inhibiting the high expression of pro-angiogenic factors, especially acting on the VEGF / VEGFR signaling pathway, is the mainstream of current research. VEGF is a glycoprotein in the human body, which plays an important role in the process of angiogenesis. The human VEGF family includes VEGF-A, VEGF-B, VEGF-C, VEGF-D, VEGF-E, and PLGF. VEGF can selectively act on VEGFR (VEGF receptor). VEGFR is a kind of tyrosine kinase transmembrane protein. After VEGF binds to VEGFR, the conformation of VEGFR changes, leading to dimerization of the receptor, and at the same time making the cell Phosphorylation occurs at the tyrosine site within and activates the downstream transduction pathway (Joukov, V., et. al. EMBO J., 1996, 15, 290-298.). A large number of studies have shown that the VEGF / VEGFR signal transduction pathway is the most important pathway for promoting angiogenesis and metastasis in cells. By inhibiting this pathway, the growth and metastasis of endothelial cells can be inhibited, thereby inhibiting the growth of tumors. At present, a number of drugs have been successfully marketed, and more than 30 drugs are in the clinical research stage. More importantly, the recombinant humanized VEGF monoclonal antibody-bevacizumab (trade name Avastin), bevacizumab is the first anti-tumor angiogenesis drug successfully marketed, which can specifically bind to VEGF-A Thereby blocking the VEGF / VEGFR pathway. This drug achieved great success in the initial stage of marketing, but with the prolongation of use time, the problem of drug resistance gradually emerged. Further studies have found that specific inhibition of VEGF-A will cause cells to release a large number of other pro-angiogenic factors, such as PLGF and FGF, and this phenomenon is called an angiogenic rescue response. In order to solve the problem of drug resistance, the development of multi-target inhibitors is a feasible strategy. Sunitinib is such a multi-target anticancer drug. Sunitinib is a multi-target tyrosine kinase inhibitor developed by Pfizer, which can effectively inhibit VEGFR-1, VEGFR-2, VEGFR-3, PDGFR-β, c-Kit, FLT-3 and other receptor tyrosine kinases, by inhibiting these proteins to block the expression of various pro-angiogenic factors in cancerous cells, to achieve the purpose of inhibiting angiogenesis and "starving" cancer cells (Abrams, T.J.et.al.Mol. Cancer Ther., 2003, 2, 1011-1021.). In addition, it also has a specific direct inhibitory effect on tumor cells with c-Kit and FLT-3 mutations. Sunitinib was approved by the FDA in 2006 and is mainly used for the treatment of gastrointestinal stromal tumor and renal cell carcinoma. It is the first anticancer drug approved for two types of indications at the same time. Although sunitinib has obvious anti-tumor effect, it still shows side effects such as fatigue, myelosuppression and fever in clinically used patients, and its tissue accumulation is strong, so it cannot be taken continuously. It is clinically used for 4 weeks and then discontinued. two week program. However, the study found that tumor neovascularization can be restored during drug withdrawal. Therefore, it is necessary to reduce toxic and side effects, optimize druggability, and find safer and more effective drugs by changing the chemical structure. Contents of the invention The purpose of the present invention is to provide a multi-target receptor tyrosine kinase inhibitor with high efficiency and low toxicity. Another object of the present invention is to provide a class of pyrrole-substituted indolinone derivatives capable of inhibiting tumor growth. Another object of the present invention is to provide a pharmaceutical composition comprising the above-mentioned pyrrole-substituted indolinone derivatives. Another object of the present invention is to provide the above-mentioned pyrrole-substituted indolinone derivatives and the use of the pharmaceutical composition containing the derivatives. The present invention provides a pyrrole-substituted indolinone derivative or a pharmaceutically acceptable salt thereof having a structure represented by the following general formula (I): in: m is selected from 0, 1, 2; n is selected from 1, 2, 3; R is selected from hydrogen, C1-C6 straight chain or branched chain alkyl, C3-C7 cycloalkyl, formyl substituted with C1-C6 straight chain or branched chain alkyl, C3-C7 cycloalkyl formyl, tert-butyl An oxycarbonyl group, a substituted carbamoyl group, or a 5-7 membered cyclic carbamoyl group. In the above-mentioned pyrrole-substituted indolinone derivatives, R is preferably selected from hydrogen, C1-C3 straight chain or branched chain alkyl, C4-C6 cycloalkyl, C1-C3 straight chain or branched chain alkyl substituted Formyl, C3-C6 cycloalkylformyl, tert-butyloxycarbonyl, N,N-dimethylformyl, N,N-diethylformyl, N,N-dipropylformyl, pyrrole Alkane-1-formyl, or piperidine-1-formyl. In the above pyrrole-substituted indolone derivatives, R is more preferably selected from hydrogen, methyl, tert-butyloxycarbonyl, N,N-dimethylformyl, or pyrrolidine-1-formyl. In the present invention, the pyrrole-substituted indolinone derivatives having the structure represented by the general formula (I) are preferably selected from the following compounds 1-15: The pharmaceutically acceptable salt of the pyrrole-substituted indolinone derivatives of the present invention is not particularly limited, and may be hydrochloride, fumarate, maleate, citrate, phosphate, sulfate, tartaric acid Salt, methanesulfonate and benzenesulfonate, etc. When hydrochloride is used, high crystallinity and high solubility can be obtained, and hygroscopicity is improved, so hydrochloride is preferably used. In the second aspect of the present invention, there is also provided a method for preparing the pyrrole-substituted indolinone derivatives of the present invention, the method comprising the following steps: (a) make 3 shown in structural formula I, 5-dimethyl-2-pyrrole carboxaldehyde and potassium nitrate concentrated sulfuric acid generation nitration reaction generates the compound shown in structural formula II: Specifically, 3,5-dimethyl-2-pyrrole carboxaldehyde represented by structural formula I was dissolved in concentrated sulfuric acid, the temperature was lowered to minus 10° C., and then potassium nitrate was added to keep the temperature for reaction. After the reaction, add cold water and stir vigorously to obtain compound II by filtration, and recrystallize to obtain pure product. (b) make 3,5-dimethyl-4-nitro-2-pyrrole carboxaldehyde shown in structural formula II and 5-fluoroindolinone shown in structural formula III undergo a condensation reaction under the catalysis of pyrrolidine to generate The compound shown in structural formula IV: Specifically, add 3,5-dimethyl-4-nitro-2-pyrrole carboxaldehyde shown in structural formula II into ethanol, heat up to 50°C, and then add 5-fluoroindolinone shown in structural formula III , insulation reaction. After the reaction, the pure compound IV was obtained by filtration. (c) make the compound shown in structural formula IV and zinc dust generation reduction reaction generate the compound shown in structural formula V: Specifically, the compound represented by the structural formula IV is dissolved in a mixed solution of tetrahydrofuran, water and methanol, the temperature is raised to 50° C., saturated ammonium chloride and zinc powder are added, and the reaction is carried out with heat preservation. After the reaction, the solvent was evaporated to dryness and extracted with ethyl acetate to obtain the pure compound V. (d) make the compound shown in structural formula V and corresponding acid VI generation condensation reaction generate the compound shown in structural formula VII: Specifically, the compound represented by the structural formula V is dissolved in tetrahydrofuran, and a base (DIPEA, DMAP, pyridine, etc.) and a condensing agent (EDCI, DCC, etc.) are added at room temperature to carry out a heat preservation reaction. After the reaction, the solvent was evaporated to dryness to obtain the crude compound represented by structural formula VII, which was washed with water and rinsed with a solvent (ethyl acetate, methanol, etc.) to obtain pure compound VII. According to the present invention, the present invention provides a pharmaceutical composition comprising one or more pyrrole-substituted indolinone derivatives of general formula (I) or a pharmaceutically acceptable salt thereof in a therapeutically effective dose, the composition can be further Including conventional pharmaceutical excipients, such as excipients, sweeteners, etc. The pyrrole substituted indolinone derivatives or pharmaceutically acceptable salts thereof of the present invention have the activity of inhibiting tyrosine kinases and can be used for preparing medicines for treating tumors caused by abnormal expression of tyrosine kinases. That is, the pyrrole-substituted indolinone derivatives of the present invention or pharmaceutically acceptable salts thereof can be used to treat tyrosine kinase-mediated tumors and inhibit the growth of related tumor cells, including administering a therapeutically effective amount of pyrrole The substituted indolinone derivatives or pharmaceutically acceptable salts thereof can also be used to prepare drugs for treating tumors mediated by tyrosine kinases and inhibiting the growth of related tumor cells. The pyrrole-substituted indolinone derivatives prepared by the invention or the pharmaceutically acceptable salts thereof have inhibitory effects on various tyrosine kinases, and whole animal experiments show that the compounds have the effect of inhibiting tumor growth. In particular, the pyrrole-substituted indolinone derivatives or pharmaceutically acceptable salts of the present invention have very low toxic and side effects. The compounds can be used to treat various tumor diseases. The compound of the invention is simple to synthesize, easy to prepare, and has abundant synthetic raw materials. The present invention will be further described below in conjunction with specific examples, but the present invention is not limited thereto. In the following preparation examples, 1H-NMR was measured with Varian Mercury AMX300, 400, and 500 instruments. MS was determined with VG ZAB-HS or VG-7070 and Esquire 3000plus-01005. All solvents were re-distilled before use, and the anhydrous solvents used were obtained by drying according to standard methods. Unless otherwise stated, all reactions were carried out under the protection of argon and tracked by TLC. The post-treatment was washed with saturated brine and dried with anhydrous magnesium sulfate. The purification of the product uses silica gel column chromatography unless otherwise specified. The silica gel used is 200-300 mesh, and GF254 is produced by Qingdao Ocean Chemical Factory or Yantai Yuanbo Silica Gel Company. Preparation Example 1: Preparation of Compound 1 The raw material 3,5-dimethyl-2-pyrrole carboxaldehyde I (5g, 40mmol) was dissolved in 60mL of concentrated sulfuric acid, then the temperature of the system was lowered to -10°C, and potassium nitrate (4.35g , 42mmol), the addition was completed in about 2h. During this process, the temperature was kept at -10°C. After the addition, the stirring was continued at this temperature for about 2 hours. After the reaction was detected by TLC, the solution was added to 1L of ice water and extracted twice with 1L of ethyl acetate. Wash with saturated brine, dry over anhydrous sodium sulfate, filter and evaporate the organic solvent to dryness under reduced pressure to obtain 7 g of crude product, which is added to 10-20 mL of ethyl acetate, stirred vigorously and filtered to obtain 5 g of pure target compound II. Compound II (1.68g, 10mmol) and compound III (1.8g, 12mmol) were added to 50mL absolute ethanol, and tetrahydropyrrole (850mg, 12mmol) was added at room temperature. After the addition, the color of the system turned yellow, and the temperature was raised to 50°C, and the reaction was continued at this temperature for 2h. After the reaction, the system was directly filtered, and the filter cake was washed with a small amount of ethanol and ethyl acetate to obtain 2.7 g of pure target compound IV. 1H NMR (400MHz, DMSO-d6) δ11.14(s, 1H), 7.88(dd, J=9.2, 2.4Hz, 1H), 7.82(s, 1H), 7.05-6.97(m, 1H), 6.88( dd, J=8.5, 4.5Hz, 1H), 2.64(s, 3H), 2.58(s, 3H). Take compound IV (900mg, 3mmol) in a 500mL two-necked flask, and then add 200mL tetrahydrofuran, 100mL methanol, 60mL water, and 60mL saturated ammonium chloride solution into it. After the addition, the temperature was raised to 50°C, then zinc powder (1.8g, 30mmol) was added under stirring, and the reaction was continued for 2h under this condition after the zinc powder was added, during which the system became clear and then cloudy. After becoming turbid, the LC-MS detection reaction was completely completed. After the reaction was complete, the solution was evaporated to dryness, and saturated sodium carbonate solution was added to make the system alkaline, and extracted twice with 2L ethyl acetate. The ethyl acetate layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic solvent was evaporated under reduced pressure to obtain the target compound V (800 mg). Dissolve compound V (270mg, 1mmol) in tetrahydrofuran (20mL), add Boc-protected 4-piperidinecarboxylic acid (270mg, 1.2mmol), EDCI (220mg, 1.1mmol), DIPEA (260mg, 2mmol) and Catalytic amount of DMAP. After the addition, the reaction was continued at room temperature for about 8 hours, and the reaction was detected by TLC. After the reaction, the tetrahydrofuran solution was evaporated to dryness, a large amount of ethyl acetate and water were added to separate the layers, and the crude product of the target compound was obtained by filtration. The crude product was washed with methanol to obtain the pure product 1. 1H NMR (400MHz, DMSO-d6) δ13.59(s, 1H), 10.83(s, 1H), 9.12(s, 1H), 7.71(dd, J=9.5 , 2.6Hz, 1H), 7.66(s, 1H), 6.93-6.86(m, 1H), 6.86-6.81(m, 1H), 3.99-3.95(m, 2H), 3.10-2.94(m, 1H), 2.79-2.75 (m, 2H), 2.17 (s, 3H), 2.15 (s, 3H), 1.82-1.78 (m, 2H), 1.55-1.45 (m, 2H), 1.41 (s, 9H). Preparation Example 2: Preparation of Compound 2 Take compound 1 (480mg, 1mmol) and add it into 10mL tetrahydrofuran, add 10mL trifluoroacetic acid at room temperature, then raise the temperature to 50°C and continue the reaction for about 2h, and the reaction is complete as detected by LC-MS. After the reaction, most of the solution was evaporated to dryness, and the rest was neutralized with saturated sodium carbonate solution and filtered to obtain the crude product. The crude product was washed with ethyl acetate and methanol to obtain pure product 2. 1H NMR (400MHz, DMSO-d6) δ13.60(s, 1H), 10.85(s, 1H), 9.26(s, 1H), 7.71(dd, J=9.5, 2.4Hz, 1H), 7.67(s, 1H), 6.94-6.86(m, 1H), 6.87-6.83(m, 1H), 3.34(d, J=12.3Hz, 2H), 3.06-2.89 (m, 2H), 2.74-2.59 (m, 1H), 2.18 (s, 3H), 2.16 (s, 3H), 2.05-1.94 (m, 2H), 1.89-1.74 (m, 2H). Preparation Example 3: Preparation of Compound 3 Take compound 2 (383mg, 1mmol) and add it to 20mL tetrahydrofuran, add DIPEA (260mg, 2mmol) and dimethylcarbamoyl chloride (214mg, 2mmol) at room temperature, continue to react for about 12h after the addition, and TLC detects that the reaction is almost complete . After the reaction, the solvent was evaporated to dryness and the solid was rinsed with 20 mL of ethyl acetate and 10 mL of methanol to obtain the pure product of the target compound 3. 1H NMR (400 MHz, DMSO-d6) δ13.58 (s, 1H), 10.82 (s, 1H), 9.10(s, 1H), 7.70(dd, J=9.5, 2.5Hz, 1H), 7.66(s, 1H), 6.95-6.86(m, 1H), 6.85-6.77(m, 1H), 3.87-3.70( m, 1H), 3.63-3.54(m, 2H), 2.86-2.58(m, 8H), 2.18(s, 3H), 2.15(s, 3H), 1.85-1.76(m, 2H), 1.71-1.55( m, 2H). Preparation Example 4: Preparation of Compound 4 The experimental operation is as the synthesis of method 1, and the Boc-protected piperidinecarboxylic acid is replaced with Boc-protected proline to obtain the target compound 4. 1H NMR (400MHz, DMSO) δ13.59 (s, 0.6H), 13.58 (s, 0.4H ), 10.84(s, 1H), 9.20(s, 0.6H), 9.13(s, 0.4H), 7.70(dd, J=9.5, 2.5Hz, 1H), 7.66(d, J=3.0Hz, 1H) , 6.89(dd, J=12.5, 5.5Hz, 1H), 6.83(dd, J=8.4, 4.7Hz, 1H), 4.38-4.15(m, 1H), 3.52-3.41(m, 1H), 3.35-3.28 (m, 1H), 2.35-2.22(m, 1H), 2.21(s, 2H), 2.18(s, 3H), 2.16(s, 1H), 1.98-1.79(m, 3H), 1.43(s, 3H) ), 1.39 (s, 6H) (Boc substituent cannot rotate freely to generate isomerism). Preparation Example 5: Preparation of Compound 5 Take Compound 2 (383mg, 1mmol) and add it to 20mL of a mixed solvent of tetrahydrofuran and methanol (1:1), add formaldehyde aqueous solution (500mg, 5mmol) and sodium cyanoborohydride (120mg, 2mmol) at room temperature, after the addition is complete Continue to react for 12h, TLC detects the reaction process, evaporates the solvent after the reaction is completed, and obtains the target compound 5 by column chromatography. 1H NMR (400MHz, DMSO-d6) δ13.58 (s, 1H), 10.83 (s, 1H), 9.05 (s, 1H), 7.70(dd, J=9.5, 2.4Hz, 1H), 7.66(s, 1H), 6.93-6.86(m, 1H), 6.85-6.80(m, 1H), 2.87-2.77(m , 2H), 2.36-2.21 (m, 1H), 2.17 (s, 3H), 2.16 (s, 3H), 2.15 (s, 3H), 1.94-1.59 (m, 6H). Preparation Example 6: Preparation of Compound 6 The synthesis method of compound 6 is the same as that of 2. Substitution of 1 with 4 gave target compound 6. 1H NMR (400MHz, DMSO-d6) δ 13.60(s, 1H), 10.83(s, 1H), 9.23(s, 1H), 7.71(dd, J=9.3, 2.4 Hz, 1H), 7.67(s, 1H), 6.93-6.87(m, 1H), 6.83(dd, J=8.4, 4.6Hz, 1H), 3.70(dd, J=8.7, 5.5Hz, 1H), 2.91 (t, J=6.6Hz, 1H), 2.18(s, 2H), 2.16(s, 2H), 2.10-1.98(m, 1H), 1.85-1.74(m, 1H), 1.72-1.63(m, 2H ). Preparation Example 7: Preparation of Compound 7 The synthesis method of compound 7 is the same as that of 1. Substituting Boc-protected 4-piperidinecarboxylic acid with Boc-protected 2-piperidinecarboxylic acid gave target compound 7. 1H NMR (400MHz, DMSO-d6) δ13.60(s, 1H), 10.84(s, 1H), 9.16( s, 1H), 7.71(dd, J=9.5, 2.5Hz, 1H), 7.67(s, 1H), 6.94-6.87(m, 1H), 6.85-6.81(m, 1H), 4.79-4.63(m, 1H), 3.87-3.75(m, 1H), 3.30-3.09(m, 1H), 2.19(s, 3H), 2.17(s, 3H), 1.81-1.59(m, 3H), 1.41(s, 9H) , 1.44-1.22 (m, 3H). Preparation Example 8: Preparation of Compound 8 The synthesis method of compound 8 is the same as that of 3. Substitution of 2 with 4 gave target compound 8. 1H NMR (400MHz, DMSO-d6) δ 13.59(s, 1H), 10.84(s, 1H), 8.99(s, 2H), 7.74-7.68(m, 1H), 7.66(s, 1H), 6.93-6.86(m, 1H), 6.83(dd, J=8.4, 4.6Hz, 1H), 4.39(t, J=7.4Hz, 1H), 3.60-3.44(m, 1H) , 3.43-3.37(m, 1H), 2.80(s, 6H), 2.28-2.20(m, 1H), 2.16(s, 3H), 2.14(s, 3H), 1.97-1.88(m, 1H), 1.87 -1.70 (m, 2H). Preparation Example 9: Preparation of Compound 9 The synthesis method of compound 8 is the same as that of 3. Substitution of 2 with 4 gave target compound 9. 1H NMR (400MHz, DMSO-d6) δ 13.59(s, 1H), 10.83(s, 1H), 8.97(s, 1H), 7.70(dd, J=9.6, 2.5 Hz, 1H), 7.66(s, 1H), 6.92-6.86(m, 1H), 6.84-6.81(m, 1H), 3.30-3.25(m, 1H), 2.99(d, J=13.2Hz, 1H) , 2.60(t, J=11.3Hz, 1H), 2.18(s, 2H), 2.16(s, 2H), 1.91-1.73(m, 2H), 1.56-1.33(m, 4H). Preparation Example 10: Preparation of Compound 10 The synthesis method of compound 8 is the same as that of 3. Substitution of 2 with 4 gave target compound 10. 1H NMR (400MHz, DMSO-d6) δ 13.59(s, 1H), 10.83(s, 1H), 9.20(s, 1H), 7.71(dd, J=9.4, 2.5 Hz, 1H), 7.67(s, 1H), 6.89(dd, J=13.8, 6.7Hz, 1H), 6.86-6.82(m, 1H), 4.13-4.02(m, 1H), 3.89(d, J= 13.1Hz, 1H), 2.94-2.72(m, 2H), 2.48-2.41(m, 1H), 2.18(s, 3H), 2.16(s, 3H), 2.01-1.94(m, 1H), 1.73-1.57 (m, 2H), 1.42 (s, 9H), 1.39-1.24 (m, 1H). Preparation Example 11: Preparation of Compound 11 Take compound 2 (383mg, 1mmol) and add it to 20mL tetrahydrofuran, add DIPEA (260mg, 2mmol) and phenyl p-nitrochloroformate (240mg, 1.2mmol) at room temperature, continue to react for about 12h after the addition, and detect the reaction by TLC almost. After the reaction, tetrahydropyrrole (142mg, 2mmol) and excess DIPEA (260mg, 2mmol) were added, and the reaction was continued for more than 12h after the addition, and the reaction was detected by TLC. After the reaction, the solvent was evaporated to dryness and the solid was rinsed with 20 mL of ethyl acetate and 10 mL of methanol to obtain the pure product 11 of the target compound. 1H NMR (400 MHz, DMSO-d6) δ13.58 (s, 1H), 10.82 (s, 1H), 9.09 (s, 1H), 7.70(dd, J=9.4, 2.5Hz, 1H), 7.66(s, 1H), 6.93-6.86(m, 1H), 6.83(dd, J=8.5, 4.6Hz, 1H), 3.70(d, J=13.5Hz, 2H), 3.27(t, J=6.4Hz, 4H), 2.73(t, J=11.6Hz, 1H), 2.18(s, 3H), 2.15(s, 3H), 1.86-1.71 (m, 6H), 1.67-1.54 (m, 2H). Preparation Example 12: Preparation of Compound 12 The synthesis method of compound 12 is the same as 2. Substitution of 1 with 10 gave target compound 12. 1H NMR (400MHz, DMSO-d6) δ 13.59(s, 1H), 10.90(s, 1H), 9.17(s, OH), 7.69(dd, J=9.4, 2.4 Hz, 1H), 7.65(s, 1H), 6.94-6.82(m, 2H), 3.67-3.52(m, 2H), 3.15-2.56(m, 3H), 2.45(d, J=10.0Hz, 2H) , 2.17(s, 3H), 2.15(s, 3H), 1.89(d, J=9.2Hz, 1H), 1.61(d, J=10.4Hz, 2H), 1.42(s, 1H). Preparation Example 13: Preparation of Compound 13 The synthesis method of compound 13 is the same as that of 3. Substitution of 2 with 12 gave target compound 13. 1H NMR (400MHz, DMSO-d6) δ 13.59(s, 1H), 10.83(s, 1H), 9.18(s, 1H), 7.71(dd, J=9.4, 2.4 Hz, 1H), 7.66(s, 1H), 6.89(dd, J=9.2, 2.4Hz, 1H), 6.86-6.80(m, 1H), 3.72-3.59(m, 1H), 3.51(d, J= 13.2Hz, 1H), 2.90-2.81(m, 1H), 2.78-2.68(m, 7H), 2.64-2.55(m, 1H), 2.18(s, 3H), 2.15(s, 3H), 1.97(d , J=14.9Hz, 1H), 1.73-1.57(m, 2H), 1.53-1.38(m, 1H). Preparation Example 14: Preparation of Compound 14 The synthesis method of compound 14 is the same as that of 11. Substituting 2 with 12 gave target compound 14. 1H NMR (400MHz, DMSO-d6) δ 13.59(s, 1H), 10.83(s, 1H), 9.18(s, 1H), 7.71(dd, J=9.4, 2.5 Hz, 1H), 7.66(s, 1H), 6.93-6.86(m, 1H), 6.83(dd, J=8.4, 4.8Hz, 1H), 3.75(d, J=12.7Hz, 1H), 3.61(d , J=12.4Hz, 1H), 3.28(s, 4H), 2.90-2.81(m, 1H), 2.73(t, J=11.4Hz, 1H), 2.60-2.50(m, 1H), 2.06-1.91( m, 1H), 1.76 (s, 4H), 1.74-1.59 (m, 2H), 1.55-1.39 (m, 1H). Preparation Example 15: Preparation of Compound 15 The synthesis method of this step is the same as that of compound 6, and the target compound is obtained by replacing the L-form N-Bn proline with the D-form N-Bn proline. 1H NMR (400MHz, DMSO) δ13.60(s, 1H), 10.83(s, 1H), 9.23(s, 1H), 7.71(dd, J=9.3, 2.4Hz, 1H), 7.67(s, 1H) , 6.93-6.87(m, 1H), 6.83(dd, J=8.4, 4.6Hz, 1H), 3.70(dd, J=8.7, 5.5Hz, 1H), 2.91(t, J=6.6Hz, 1H), 2.18 (s, 2H), 2.16 (s, 2H), 2.10-1.98 (m, 1H), 1.85-1.74 (m, 1H), 1.72-1.63 (m, 2H). Preparation Example 16: Preparation of Compound 6 Hydrochloride Take 0.5 mL of saturated ethanol hydrogen chloride solution, dilute it ten times with absolute ethanol, add compound 6 (368 mg, 1 mmol), stir for 5 to 10 minutes, concentrate the reaction solution under reduced pressure, and wash with a small amount of methanol to obtain the compound 6 hydrochloride. The hydrochloride salts of all other compounds can be prepared by reacting the corresponding compounds with dilute hydrochloric acid ethanol solution in this way. The above preparation examples of pyrrole-substituted indolinone derivatives are for reference, and other such derivatives can also be prepared by referring to the above-mentioned methods. In addition, the applicant synthesized the following comparative compounds 1-3 by a method similar to the above or other known methods in the art. Comparative compound 1 is the same as compound 2, the difference is that the piperidinyl group on the far right is connected to the carbonyl group by N atom Comparative compound 2 is the same as compound 6, the difference is that the pyrrolidinyl group on the far right is connected to the carbonyl group through N atom. Comparative compound 3 is the same as compound 2, except that the piperidinyl group on the far right is connected to the carbonyl group through a methylene group. Example The present invention will be further described below in conjunction with specific examples, but these examples should not be construed as limiting the present invention. Test Example 1: Determination of KDR Tyrosine Kinase Biochemical Activity in Vitro The in vitro inhibitory activity of compounds on KDR (VEGF receptor) tyrosine kinase was determined by HTRF (homogeneous time-resolved fluorescence) method. Add the mixture of kinase buffer, test compound or sunitinib, substrate and ATP solution to a final volume of 10 μL to a 384-well plate, and incubate at room temperature for an appropriate time; add 10 μL SA-XL665 and TK antibody to each well, and incubate at room temperature for 1 h , read with Synergy2. The results showed that the compounds of the above examples had significant inhibitory activity on KDR at concentrations of 0.1 μM and 1 μM. The activities of compounds 3, 6, 8, 9, 11, 13, etc. were comparable to those of sunitinib. The in vitro inhibitory activity of table 1 embodiment compound to KDR IC50(nM) 2 290 3 78 5 141 6 83 8 66 9 89 11 74 12 142 13 77 14 143 15 65 Sunitinib 62 Staurosporine 8.14 Staurosporine (staurosporine): positive control compound Test Example 2: Determination of Cytotoxicity to HUVEC and VEGF-induced Proliferation Activity of HUVEC Cells in Vitro Detection of VEGF-induced proliferation inhibitory activity of human umbilical vein endothelial cells (HUVEC): Human umbilical vein endothelial cells (HUVEC) were cultured in F-12K containing 10% FBS, 18u / mL heparin and 30 μg / mL ECGS. 4-8 generations of HUVEC. Cells were digested with trypsin, resuspended in culture medium (1×105 / mL), 100 μL per well was added to a 96-well plate, and adhered overnight. Replace the F-12K medium containing 5% FBS and cultivate for 24 hours. Add test compound or sunitinib or control 5% FBS-F-12K culture solution and incubate for 30min. Add 0.1% FBS-F-12K culture solution with a final concentration of 30ng / mL VEGF165 or vehicle (DMSO), and induce culture for 72 hours. Aspirate the culture medium, add 120 μL MTS detection solution to each well, incubate at 37°C, and read OD490. The 5% FBS-F-12K culture solution treatment group was used as the negative control, and the OD value of the negative control group was subtracted from the OD value of the VEGF165 stimulation group to obtain the VEGF-stimulated growth value, and the inhibition rate was calculated, and the dose-effect curve was prepared with GraphPad Prism software and Calculate the median effect concentration (EC50). Cytotoxicity test: the above HUVEC cells were cultured in F-12K medium containing 10% fetal bovine serum (FBS), 100U / ml penicillin, 100μg / mL streptomycin, 30ug / mL ECGS, 18u / mL heparin, trypsin Digest HUVEC in the logarithmic growth phase, adjust the cell density to the required concentration with F-12K complete medium containing 5% FBS, inoculate 150 μL of cells in a 96-well plate, 3000 / well, add 5% FBS after 24 hours 50 μL of the 4-fold concentration of the test compound diluted in complete culture medium was used as a control with the same volume of DMSO dilution. After the cells were cultured for 72 hours, 20 μL MTS and 1 μl PMS were added to each well. Measure OD490 after 1-2 hours, and use OD650 value as a reference. GraphPad Prism software was used to make dose-response curves and calculate the half cytotoxic concentration (CC50). Calculate the therapeutic index (Therapeutic Index, TI) of the test compound on human umbilical vein endothelial cells (HUVEC), TI=CC50 / EC50. The results show that the compounds of the above examples can significantly inhibit the proliferation of HUVEC cells stimulated by VEGF, but the activity is weaker than that of sunitinib. However, the cytotoxicity of some compounds (compounds 2, 3, 5, 6, 8, 11, 13, 14, 15) on HUVEC was significantly lower than that of sunitinib. The therapeutic index (TI) of compounds 2, 3, 5, 6, 8, 11, 14, and 15 is about 2-3 times that of sunitinib, showing a larger therapeutic window. In comparative compounds 1 and 2, since the rightmost nitrogen-containing heterocyclic group is connected to the carbonyl group through a heteroatom, its therapeutic index (TI) is roughly equivalent to that of sunitinib, which is significantly lower than that of the compound of the present invention. In comparative compound 3, since the nitrogen-containing heterocyclic group on the far right is connected to the carbonyl through a methylene group, its therapeutic index (TI) is also roughly equivalent to that of sunitinib, which is significantly lower than that of the compound of the present invention Table 2. Cytotoxicity, VEGF-induced in vitro proliferation activity and therapeutic index of some compounds on HUVEC Compound EC50 (NM) CC50 (NM) Ti = CC50 / EC50 2 16.15> 20000> 1238 3 13.74> 20000> 1456 5 18.43> 20000> 1085 6 13.35 17566.77 1316 8 11.35> 1762 11 13.93> 143666666666666666666666666666666 14 13.04 >20000 >1534 15 14.21 17732.12 1247 Sunitinib 7.73 4144.09 536 Comparative Compound 1 13.23 5689.26 430 Comparative Compound 2 12.31 6982.25 567 Comparative Compound 3 14.64 8054. Test Example 3: Determination of Inhibitory Proliferation Activity of Human MV-4-11 Tumor Cell Line Human acute leukemia cell MV-4-11 is a Flt-3 mutant cell line. The in vitro antiproliferative activity of the compound against MV-4-11 was determined by the MTS method: cells in the logarithmic phase of growth were digested with trypsin, counted, an appropriate amount of cells were resuspended in culture medium, and 150 μL per well was added to a 96-well plate. After overnight incubation. Add 50 μL of 4-fold serially diluted test compound or control culture solution to each well, and incubate for 72 hours. Aspirate the culture solution, add 120 μL MTS detection solution (100 μL fresh medium and 20 μL MTS solution) to each well, incubate at 37°C, and read the OD490 value. Graphpad Prism5 software was used to analyze and process the data to obtain IC50. The results showed that the compounds 1-15 of the above examples all showed significant anti-proliferation activity against MV-4-11, and the activity of some compounds was equivalent to or stronger than that of sunitinib (see the table below). FLT-3 (FMS-like tyrosine kinase 3) is a type III receptor tyrosine kinase widely distributed in systemic, immune and nervous systems. FLT-3 gene mutation and overexpression will lead to tumorigenesis. The specific anti-proliferative activity of compounds 1-15 on MV-4-11 also suggested that the compounds of the examples were FLT-3 inhibitors similar to sunitinib. Table 3. The inhibitory effect of some compounds on the proliferation of human MV-4-11 cell lines in vitro Compound IC50(nM) Maximum inhibition rate(%) 2 4.70 92.6 3 10.67 92.1 5 1.68 95.0 6 11.58 92.4 9 5.34 94.3 12 7.87 93.1 13 6.41 90.2 15 12.34 92.5 494 Sunitinib 9 3. Test Example 4: Inhibition of MV-4-11 Nude Mice Transplanted Tumors in Vivo MV-4-11 cells were cultured and expanded in vitro, the cells in the logarithmic growth phase were collected, resuspended in serum-free EMEM medium, and the cell suspension was injected subcutaneously into the axilla of the anterior right limb of male Balb / c nude mice with a syringe. Regularly observe the growth of the animals and transplanted tumors; when the tumor volume grows to about 100-300mm3, select animals with moderate tumor size and divide them into random groups, with 6 animals in each group. Give blank vehicle (0.5% CMC) or the suspension of above-mentioned embodiment compound 6 or Sunitinib respectively, intragastric administration, dosage is 80mg / kg, once a day, administration cycle 3 weeks; During administration, monitor The diameter of the tumor, the weight of the animal, and the living conditions of the animal were observed; the experiment was terminated after 3 weeks of administration, and the animal was sacrificed with CO2 and dissected. The calculation formula of tumor volume (TV) is: TV=1 / 2×a×b2, wherein, a represents the long diameter of the tumor; b represents the short diameter of the tumor. The results showed that after intragastric administration for 21 days, the tumor in the solvent control group increased to nearly 6 times the initial volume, while the transplanted tumor in the compound 6 treatment group completely disappeared, and the compound 6 had no significant effect on the body weight of the animals. Although sunitinib also showed obvious anti-tumor effect, most of the animal tumors disappeared, but the animal body weight decreased significantly, and the toxicity was obvious. Table 4. The inhibitory effect of compound 6 on MV-4-11 nude mouse xenograft tumor **: P<0.01, compared with solvent control From MV-4-11 nude mice transplanted tumor experimental results, it can be found that compound 6 of the present invention has a good inhibitory effect on MV-4-11 transplanted tumor, and the 80mg / kg dose can make the tumor disappear completely, while the effect on body weight The effect of sunitinib was small, but the body weight of the animals was significantly decreased by sunitinib, and the toxicity was obvious. The results show that this series of compounds have anti-tumor effects comparable to sunitinib, but with less toxicity, larger therapeutic window, and better development value.

Claims

------15 / 12 / 2017------(OCR)Page 1 of 6 Claim 1. Indolone derivatives which are replaced by -pyrrole with a structure which is shown in the general formula (I) below, compounds 4, 6, 7, 8, 9, 15, or pharmaceutically acceptable salts of those: (chemical formula) where m is chosen from 0, 1 and 2; n is chosen from 1, 2 and 3; and R is chosen from hydrogen, C1-C6 linear or branched alkyl, C3-C7 cycloalkyl, formyl which are replaced by C1-C6 linear or branched alkyl, C3-C7 cycloalkylformyl, t-butoxycarbonyl, carbamoyl which are replaced, or cyclic carbamoyl with members -5 to -7, (chemical formula)(chemical formula)Page 2 of 6(chemical formula)Page 3 of 62.Indolone derivatives that are substituted for pyrrole pursuant to claim 1, where R is chosen from hydrogen, C1-C3 linear or branched alkyl, C4-C5 cycloalkyl, formils that are substituted for C1-C3 linear or branched alkyl, C3-C6 cycloalkyl formil, t-butoxycarbonyl, N,N-dimethylcarbamoyl, N,N-chiethylcarbamoyl, N,N-dipropylcarbamoyl, pyrolidin-1-ylformyl, or piperidin-1-ylformyl3. Indolone derivatives that are substituted for pyrrole pursuant to claim 1, where the child R is chosen from hydrogen, methyl, t-butoxycarbonyl, N,N-dimethylcarbamoyl, or pyrolidin-1-ylformyl4. An indolone derivative which is replaced by pyrrole pursuant to claim 1, where an indolone derivative which is replaced by pyrrole with a structure which is shown in general formula(I) is selected from compounds 1 to 3, 5, 10 to 14 below: (chemical formula) page 4 of 6 pages (chemical formula) page 5 of 6 pages or 14 chemical formulas.

5. An indolone derivative which is replaced by pyrrole or a pharmaceutically acceptable salt of those pursuant to any one of claims 1 to 4, where the salt is a hydrochloride.6.Pharmaceutical components, comprising: therapeutic amounts of one or more indolone derivatives which are replaced by pyrrole or pharmaceutically acceptable salts of them under any of the claims 1 through 5, and as an optional auxiliary; 7. The use of indolone derivatives which are replaced by pyrrole or pharmaceutically acceptable salts of them under any of the claims 1 through 5 or the pharmaceutical component under claim 6 in the production of tyrosine kinase inhibitors; 8. The use of indolone derivatives which are replaced by pyrrole or pharmaceutically acceptable salts of them under any of the claims 1 through 5 or the pharmaceutical component under claim 6 in medicines for the treatment and / or prevention of diseases associated with tyrosine kinase receptors in mammals; 9.The use of incolon derivatives replaced by pyrrole or their pharmaceutically acceptable salts under any of the claims 1 through 5 or the pharmaceutical component under claim 6 in medicines for the treatment or as an adjunct and / or prevention of (i) tumors mediated by tyrosine kinase receptors or (ii) tumor cell proliferation and migration driven by tyrosine kinase receptors, in mammals.

10. The use under claim 8 or 9, where the mammal is human.

11. The method of the treatment and / or prevention of tyrosine kinase-associated tumors in mammals comprising the administration of an effective therapeutic dose of incolon derivatives replaced by pyrrole or their pharmaceutically acceptable salts under any of the claims 1 through 5 or the pharmaceutical component under claim 6.12.Methods of treatment or complementary and / or prevention of (i) tumors mediated by tyrosine kinase receptors or (ii) tumor cell proliferation and 6-6-6 migration driven by tyrosine kinase receptors, in mammals, which are comprised of the administration of an effective therapeutic dose of an ondolone derivative substituted for pyrrole or a pharmaceutically acceptable salt of such as any of Claims 1 through 5 or a pharmaceutical component as per Claims 613. Methods as per Claims 11 or Claims 12, where the mammal is human------------1.Indolone derivatives which are replaced by -pyrrole with the structure which is shown in the general formula(I) below, compounds 4, 6, 7, 8, 9, 15, or pharmaceutically acceptable salts of such:(chemical formula)(I) where m is chosen from 0, 1 and 2; n is chosen from 1, 2 and 3; and R is chosen from hydrogen, C1-C6 linear or branched alkyl, C3-C7 cycloalkyl, formyl which is replaced by C1-C6 linear or branched alkyl, C3-C7 cycloalkylformyl, t-butoxycarbonyl, carbamoyl which is replaced, or cyclic carbamoyls with members -5 to -7,4 (chemical formula), 6 (chemical formula), 7 (chemical formula), 8 (chemical formula), 9 (chemical formula), 15 (chemical formula).

2. Indolone derivatives which are replaced by -pyrrol pursuant to claim 1, where R is chosen from hydrogen, C1-C3 linear alkyl or branched C4-C6 cycloalkyl, formyl which is replaced by C1-C3 linear alkyl or branched, C3-C6 cycloalkylformyl, t-butoxycarbonyl, N,N-dimethylcarbamoyl, N,N-diethylcarbamoyl, N,N-dipropylcarbamoyl, pyrroldin-1-ylformyl, or piperidin-1-ylformyl. 3.Indolone derivatives that are replaced by pyrrole pursuant to claim 1, where R is chosen from hydrogen, methyl, t-butoxycarbonyl, N,N-dimethylcarbamoyl, or pyrolidin-1-ylformyl4. Indolone derivatives that are replaced by pyrrole pursuant to claim 1, where the indolone derivatives that are replaced by pyrrole with the structures shown in general formula(I) are chosen from compounds 1 to 3, 5, 10 to 14 below: 1(chemical formula)2(chemical formula)3(chemical formula)5(chemical formula)10(chemical formula)11(chemical formula)12(chemical formula)13(chemical formula) 1. Chemical formula (chemical formula) or 14 (chemical formula) 5. Indolone derivatives which are replaced by -pyrrole or any pharmaceutically acceptable salts of those under any of the claims 1 through 4, where the remainder are hydrochlorides.

6. Pharmaceutical composition, which includes: therapeutic amount of one or more indolone derivatives which are replaced by -pyrrole or any pharmaceutically acceptable salts of those under any of the claims 1 through 5, and, as an option, an auxiliary. 7.The use of indolone derivatives substituted for pyrrole or their pharmaceutically acceptable salts under any of the claims 1 through 5 or the pharmaceutical components under claim 6 in the manufacture of tyrosine kinase inhibitors.

8. The use of indolone derivatives substituted for pyrrole or their pharmaceutically acceptable salts under any of the claims 1 through 5 or the pharmaceutical components under claim 6 in medicines for the treatment and / or prophylactic use associated with tyrosine kinase receptors in animals.

9. Use of indolone derivatives substituted for -pyrrole or pharmaceutically acceptable salts of those under any of the claims 1 through 5 or the pharmaceutical components under claim 6 in medicines for the treatment or as an adjunct and / or prevention of (i) tumors mediated by -tyrosine kinase receptors or (ii) tumor cell proliferation and migration driven by tyrosine kinase receptors, in mammals.

10. Use under claim 8 or 9, where the mammal is human;