A bicyclic compound as a VEGFR-2 inhibitor and its application

Abstract

The present invention relates to a bicyclic compound as a VEGFR-2 inhibitor and its application. The compound structure is as shown in Formula I, or an optical isomer, isotope derivative or a pharmaceutically acceptable salt of the compound, or a pharmaceutical composition comprising the compound. The compound or pharmaceutical composition is used as a VEGFR-2 inhibitor for treating related diseases, and can be used alone or in combination with other therapeutic substances.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and in particular relates to a bicyclic compound as a VEGFR-2 inhibitor and application thereof. Background Art

[0002] Angiogenesis is also known as the formation of new blood vessels. The growth and metastasis of solid tumors require blood vessels to supply them with a large amount of oxygen and nutrients to meet their continued division and proliferation. It can be said that tumor blood vessels are channels for tumor nutrient delivery and tumor cell escape. Tumor angiogenesis is an extremely important sign of solid tumor growth, infiltration and metastasis. Anti-tumor angiogenesis drugs can degenerate existing tumor blood vessels, cut off the supply of oxygen and other nutrients required for tumor cell growth, inhibit tumor angiogenesis, and continuously inhibit tumor cell growth and metastasis. They have been used to treat many types of solid tumors.

[0003] Angiogenesis is regulated by a variety of growth factors. Among these growth factors, vascular endothelial growth factor (VEGF) is a highly specific vascular endothelial mitogen with a molecular weight of 34-45kDa. It has a strong stimulating effect on the proliferation of endothelial cells and is one of the most active and specific angiogenic factors known to date. The VEGF family mainly includes VEGF-A, -B, -C, -D, -E and placental growth factor (PIGF). Among them, VEGF-A is closely related to angiogenesis. VEGF binds to the high-affinity vascular endothelial growth factor receptor (VEGFR), thereby inducing VEGFR to form dimers, causing conformational changes in its intracellular domain, exposing the binding site for adenosine triphosphate, causing autophosphorylation of the tyrosine residues of VEGFR, and transmitting signals to the downstream, thereby promoting the proliferation of vascular endothelial cells and causing angiogenesis.

[13] . The VEGFR family has five members, of which three major subtypes belong to the tyrosine kinase receptor superfamily (RTKs), namely VEGFR-1, VEGFR-2 and VEGFR-3. VEGFR-2 is the main regulatory receptor that stimulates endothelial cell migration, proliferation, survival and increases vascular permeability. Abnormal upregulation of VEGFR-2 can serve as an important marker for many diseases, especially various malignant tumors. VEGFR-2 inhibitors are a class of small molecule inhibitors that bind to the ATP binding site of the intracellular terminal kinase domain of the VEGFR-2 receptor, block its phosphorylation, inhibit downstream signal transduction, and thus prevent VEGFR-2-dependent angiogenesis.

[0004] Since VEGFR-2 is expressed at a low level in healthy tissue cells, but highly expressed in solid tumors such as lung cancer, liver cancer, kidney cancer, non-small cell lung cancer, thyroid cancer, ovarian cancer, breast cancer, gastric cancer and colorectal cancer, the design and development of VEGFR-2 inhibitors can effectively inhibit tumor angiogenesis and thus exert anti-tumor effects, while having less toxic side effects on healthy tissues. Summary of the invention

[0005] In the first aspect of the present invention, the present invention provides a compound represented by the general structural formula I, an optical isomer, an isotopic derivative or a pharmaceutically acceptable salt thereof,

[0006]

[0007] Wherein, R1 is selected from H, halogen, C 1-3 Alkyl, C 1-3 Alkoxy or C 1-3 Alkylamino;

[0008] R2 is selected from H, halogen, C 1-3 Alkyl, C 1-3 Alkoxy or C 1-3 Alkylthio;

[0009] R3 is selected from H or halogen;

[0010] X is selected from C or N.

[0011] Furthermore, the R1 is preferably H or halogen

[0012] The R2 is preferably H, halogen or C 1-3 alkyl;

[0013] X is C.

[0014] In another aspect of the present invention, the compound of the present invention is selected from:

[0015]

[0016] The information of the compound is as follows:

[0017]

[0018]

[0019]

[0020] In the second aspect of the present invention, the present invention also provides a pharmaceutical composition comprising any compound described in the present invention, its optical isomers, isotopic derivatives or pharmaceutically acceptable salts thereof and one or more pharmaceutically acceptable excipients or carriers.

[0021] Furthermore, the pharmaceutically acceptable carrier may include a filler, a glidant, a granulation binder, a lubricant, a disintegrant, and the like.

[0022] Furthermore, the fillers include but are not limited to starches, sugars, celluloses and inorganic salts.

[0023] In some embodiments, the filler used is microcrystalline cellulose.

[0024] Furthermore, the disintegrant includes but is not limited to sodium carboxymethyl starch, sodium carboxymethyl cellulose, cross-linked sodium carboxymethyl cellulose and cross-linked polyvinylpyrrolidone.

[0025] In the third aspect of the present invention, the present invention also provides the use of the aforementioned compound, its optical isomer, isotope derivative or pharmaceutically acceptable salt or the aforementioned pharmaceutical composition in the preparation of VEGFR-2 inhibitors.

[0026] In the fourth aspect of the present invention, the present invention also provides the use of the aforementioned compound, its optical isomer, isotope derivative or its pharmaceutically acceptable salt or the aforementioned pharmaceutical composition in the preparation of drugs for treating VEGFR-2 kinase-mediated related diseases.

[0027] Furthermore, the VEGFR-2 kinase-mediated related diseases are selected from tumor diseases.

[0028] Furthermore, the tumor disease is lung cancer, liver cancer, kidney cancer, non-small cell lung cancer, thyroid cancer, ovarian cancer, breast cancer, gastric cancer or colorectal cancer.

[0029] Furthermore, the pharmaceutically acceptable salts described in the present invention are bisulfate, hydrochloride, hydrobromide, sulfate, oxalate, lactate, gluconate, tartrate, fumarate, methanesulfonate, ethanesulfonate, benzenesulfonate, acetate, citrate, and p-toluenesulfonate.

[0030] Definition and Description

[0031] Unless otherwise specified, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be considered ambiguous or unclear without a special definition, but should be understood according to the ordinary meaning.

[0032] The term "pharmaceutically acceptable" refers to those compounds, materials, compositions and / or dosage forms which, within the scope of sound medical judgment, are suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0033] The compounds of the present invention may exist in specific stereoisomeric forms. The present invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic mixtures and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the present invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl. All of these isomers and their mixtures are included within the scope of the present invention.

[0034] The present invention also includes all suitable isotopic variants of the compounds of the present invention. In this case, an isotopic variant of a compound of the present invention is understood to be a compound in which at least one atom in the compound of the present invention is replaced by another atom having the same atomic number but an atomic mass different from the atomic mass common or predominantly present in nature. Examples of isotopes that can be introduced into the compounds of the present invention are isotopes of hydrogen, carbon, nitrogen, oxygen, sulfur, fluorine, chlorine, bromine and iodine. Specific isotopic variants of the compounds of the present invention (such as those in which one or more radioactive isotopes are introduced in particular) can be used, for example, to study the mechanism of action or distribution of active substances in the body; because they can be prepared and detected relatively easily, they can be used to detect the presence of radioactive isotopes in the body. 3 H-or 14 C-isotope-labeled compounds are particularly suitable for this.

[0035] Unless otherwise specified, the term “C 1-3 "Alkyl" is used to denote a straight or branched chain saturated hydrocarbon group consisting of 1 to 3 carbon atoms.

[0036] Unless otherwise specified, the term “C 1-3 "Alkoxy" denotes those alkyl groups containing 1 to 3 carbon atoms which are linked to the rest of the molecule via an oxygen atom.

[0037] Unless otherwise specified, the term “C 1-3 The term "alkylamino" refers to 1-3 Alkyl-NH- structure group, where "C 1-3 "Alkyl" is as defined above.

[0038] Unless otherwise specified, the term “C 1-3 "Alkylthio" refers to a C 1-3 Alkyl-S-structured groups, where "C 1-3 "Alkyl" is as defined above.

[0039] Unless otherwise specified, the term "halogen" includes, for example, a fluorine atom, a chlorine atom, a bromine atom and an iodine atom.

[0040] Unless stereochemistry is explicitly indicated in a chemical structure or chemical name, the chemical structure or chemical name shall include all possible stereoisomers, conformers, rotational isomers and tautomers of the compound. For example, a compound containing a chiral carbon atom shall include the (R) enantiomer and the (S) enantiomer and mixtures of enantiomers, including racemic mixtures; a compound containing two chiral carbon atoms shall include all enantiomers and diastereomers (including (R, R), (S, S), (R, S) and (R, S) isomers).

[0041] In all uses of compounds of the formulae described herein, the present invention also includes the use of any or all stereochemical forms, enantiomers, diastereomers, conformers, rotamers, tautomers, solvates, hydrates, polymorphs, crystalline forms, amorphous forms, salts, pharmaceutically acceptable salts, metabolites, and prodrugs of the compounds.

[0042] The pharmaceutical composition of the present invention contains at least one compound according to any embodiment disclosed herein (including pharmaceutically acceptable salts of these compounds) mixed with at least one pharmaceutically acceptable excipient, carrier or diluent. Preferably, the pharmaceutical composition is a sterile composition, or a composition consisting mainly of or only of the above-mentioned compound and one or more pharmaceutically acceptable excipients, carriers and / or diluents. In some embodiments, the pharmaceutical composition comprises at least two pharmaceutically acceptable carriers and / or excipients described herein.

[0043] Certain compounds of the present invention may exist in non-solvated forms as well as solvated forms (i.e., solvates). The compounds of the present invention may also include hydrated forms (i.e., hydrates). Typically, solvates and hydrate forms are identical to non-solvated forms in terms of biological efficacy and are included within the scope of the present invention. The present invention also includes all polymorphs, including crystalline and non-crystalline forms. Typically, for the intended use of the present invention, all physical forms are available and are intended to be included within the scope of the present invention.

[0044] The compounds of the present invention may have systemic and / or local effects. For this purpose, they may be administered in a suitable manner, for example, by oral, parenteral, pulmonary, intranasal, sublingual, lingual, buccal, rectal, transdermal, transconjunctival or otic routes, or as implants or stents.

[0045] Suitable for oral administration are dosage forms which act according to the prior art, release the compound according to the invention quickly and / or modified and contain the compound according to the invention in crystalline and / or non-crystalline and / or dissolved form, such as tablets (uncoated or coated tablets, for example with an enteric coating or with a coating which delays dissolution or an insoluble coating which controls the release of the compound according to the invention), tablets which disintegrate rapidly in the mouth, or films / wafers, films / lyophilisates, capsules (for example hard or soft capsules), dragees, granules, pellets, powders, emulsions, suspensions, aerosols or solutions.

[0046] Parenteral administration can be performed by bypassing an absorption step (e.g., intravenous, intraarterial, intracardiac, intraspinal or intralumbar) or including absorption (e.g., intramuscular, subcutaneous, intradermal, transdermal or intraperitoneal). Suitable dosage forms for parenteral administration include injection and infusion preparations in the form of solutions, suspensions, emulsions, lyophilizates or sterile powders.

[0047] Dosage forms suitable for other routes of administration are, for example, inhalation forms (including powder inhalers, nebulizers), nasal drops, solutions and sprays; tablets, films / wafers or capsules for lingual, sublingual or buccal administration; suppositories, ear or eye preparations, vaginal capsules, aqueous suspensions (lotions, shaking mixtures), lipophilic suspensions, ointments, creams, transdermal therapeutic systems (e.g. patches), milks, pastes, foams, dusting powders, implants or stents.

[0048] As used herein, the term "treatment" refers to the administration of one or more drug substances, particularly compounds of formula (I) and / or pharmaceutically acceptable salts thereof, to an individual suffering from a disease or symptoms of the disease, in order to cure, alleviate, mitigate, alter, cure, improve, ameliorate or affect the disease or symptoms of the disease. As used herein, the term "prevention" refers to the administration of one or more drug substances, particularly compounds of formula (I) and / or pharmaceutically acceptable salts thereof, to an individual with a physique susceptible to the disease, in order to prevent the individual from suffering from the disease. When referring to a chemical reaction, the terms "treating", "contacting" and "reacting" refer to the addition or mixing of two or more reagents under appropriate conditions to produce the indicated and / or desired product. It should be understood that the reaction that produces the indicated and / or desired product may not necessarily result directly from the combination of the two reagents initially added, that is, there may be one or more intermediates generated in the mixture, which ultimately lead to the formation of the indicated and / or desired product.

[0049] The compounds defined in the present invention or their pharmaceutically acceptable salts, or pharmaceutically acceptable compositions containing them, are effective inhibitors of VEGFR-2. The compounds of the present invention are expected to be potentially useful agents in treating diseases or medical conditions mediated by VEGFR-2 kinase-related pathways.

[0050] The compounds according to the invention are preferably suitable for the treatment and / or prevention of diseases mediated by VEGFR-2 kinase-related pathways.

[0051] Beneficial Effects

[0052] The present invention provides a new class of organic small molecule compounds containing a bicyclic structure, which can exhibit significant inhibitory activity on VEGFR-2 kinase and show the potential for treating VEGFR-2 kinase-mediated related diseases, such as cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0053] Figure 1 The synthetic route of the intermediate and the four compounds in Example 2

[0054] Figure 2 The dose-inhibition rate curve of the four compounds in Example 2 on VEGFR-2 DETAILED DESCRIPTION

[0055] The present application is described in detail below by way of examples, but it is not intended that there is any adverse limitation to the present application. The present application has been described in detail herein, and its specific embodiments are also disclosed therein. It will be apparent to those skilled in the art that various changes and modifications can be made to the specific embodiments of the present application without departing from the spirit and scope of the present application.

[0056] The raw materials and reagents used in the following examples are all commercially available analytically pure or chemically pure drugs, and the nuclear magnetic resonance hydrogen spectra of the compounds are measured using Bruker ARX-400. Figure 1 , intermediates and synthetic routes of Examples 1-4.

[0057] Example 1 Preparation of key intermediates

[0058] 1. Preparation of intermediate mz-1

[0059] 4-Chloro-6,7-dimethoxyquinoline (5.000 g, 22.36 mmol), 4-nitrophenol (6.220 g, 44.71 mmol) and DIPEA (7.79 mL, 44.71 mmol) were added to 50 mL of chlorobenzene and refluxed for 24 hours. The reaction was complete when detected by TLC. The solvent was evaporated and water was added. The mixture was stirred for 0.5 h and filtered to obtain 6.990 g of a yellow solid, namely, intermediate mz-1, with a yield of 96%. 1HNMR (400MHz, DMSO) δ8.61 (d, J = 5.1Hz, 1H), 8.35 (s, 1H), 8.33 (s, 1H), 7.45 (s, 1H), 7. 45(s,1H),7.42(s,1H),7.37(s,1H),6.87(d,J=5.1Hz,1H),3.96(s,3H),3.88(s,3H).

[0060] 2. Preparation of intermediate Dz-1

[0061] 4-Chloro-6,7-dimethoxyquinoline (5.000 g, 22.36 mmol), 2-fluoro-4-nitrophenol (7.020 g, 44.71 mmol) and DIPEA (7.79 mL, 44.71 mmol) were added to 50 mL of chlorobenzene and refluxed for 24 hours. The reaction was complete when detected by TLC. The solvent was evaporated, water was added, and the mixture was stirred for 0.5 h. 7.390 g of yellow solid was obtained by suction filtration, i.e., intermediate Dz-1, with a yield of 96%. 1 HNMR(400MHz,DMSO)δ8.57(d,J=5.1Hz,1H),8.46(dd,J=10.4,2.7Hz,1H),8.20(d,J=8.9Hz ,1H),7.62(t,J=8.6Hz,1H),7.45(s,2H),6.78(d,J=5.1Hz,1H),3.96(s,3H),3.92(s,3H).

[0062] 3. Preparation of intermediate mz-2

[0063] Add mz-1 (8.50 g, 26.05 mmol) and reduced iron powder (6.50 g, 169.32 mmol) to 150 mL of 90% ethanol water, then add 4 mL of glacial acetic acid, reflux for 4 hours, and after TLC detection, filter while hot after the reaction is complete. Decolorize the filtrate with activated carbon, evaporate to dryness, add water and stir, and filter to obtain 2.800 g of brown solid, i.e., intermediate mz-2, with a yield of 36%. 1 HNMR(400MHz,DMSO)δ8.43(d,J=5.4Hz,1H),7.50(s,1H),7.36(s,1H),6.92(d,J=8 .7Hz,2H),6.66(d,J=8.7Hz,2H),6.37(d,J=5.3Hz,1H),5.17(s,2H),3.93(s,6H).

[0064] 4. Preparation of intermediate Dz-2

[0065] Dz-1 (8.200 g, 23.82 mmol) and reduced iron powder (8.650 g, 154.81 mmol) were added to 150 mL of 90% ethanol water, followed by 4 mL of glacial acetic acid. The mixture was refluxed for 4 hours. After the reaction was completed by TLC, it was filtered while hot (with diatomaceous earth pad). The filtrate was decolorized with activated carbon, evaporated to dryness, and stirred with water. 5.430 g of brown solid was obtained by filtration, i.e., intermediate Dz-2, with a yield of 72%. 1 H NMR (400MHz, DMSO) δ8.45(d,J=5.2Hz,1H),7.50(s,1H),7.38(s,1H),7.07(t,J=9.0Hz,1H),6.55 (dt,J=13.1,1.7Hz,1H),6.46(d,J=8.7Hz,1H),6.39(d,J=5.2Hz,1H),5.49(s,2H),3.94(s,6H).

[0066] 5. Preparation of intermediate mz-3

[0067] mz-2 (2.800 g, 9.45 mmol), 2-chloronicotinic acid 3-methyl carboxylate (2.570 g, 24.28 mmol) and cesium carbonate (6.850 g, 21.01 mmol) were added to an ethanol-toluene mixed solvent (60 mL + 30 mL), followed by palladium acetate (0.212 g, 0.944 mmol) and PdCl2 (dppf)-dcm (0.772 g, 0.94 5mmol), N2 was replaced three times, and the reaction was carried out at 110°C for 3 hours. After the reaction was completed by TLC, it was cooled, water was added, and dichloromethane was added for extraction. After filtration to remove insoluble matter, the organic layer was separated, and the aqueous layer was extracted twice with dichloromethane. The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness to obtain a black oil. Anhydrous ether was added and stirred to solidify, and 1.810g of brown solid was obtained by filtration, i.e., intermediate mz-3, with a yield of 44%. 1 H NMR (400MHz, DMSO) δ10.21(s,1H),8.50–8.42(m,2H),8.29(dd,J=7.7,2.3Hz,1H),7.86(dd,J=8.9,2.4Hz,2H),7.53(s,1H),7.40(s ,1H),7.25(d,J=6.6Hz,2H),6.92(t,J=5.1Hz,1H),6.48(d,J=5.5Hz,1H),4.38(q,J=7.2Hz,2H),3.94(s,6H),1.37(t,J=7.2Hz,3H).

[0068] 6. Preparation of intermediate Dz-3

[0069] Dz-2 (1.100 g, 3.50 mmol), methyl 2-chloronicotinic acid 3-formate (1.540 g, 8.99 mmol) and cesium carbonate (2.55 g, 7.84 mmol) were added to an ethanol-toluene mixed solvent (20 mL + 10 mL), followed by palladium acetate (0.078 g, 0.35 mmol) and PdCl2 (dppf) -dcm (0.286 g, 0.35 mmol). After N2 substitution three times, the reaction was carried out at 110 ° C for 3 hours. After the reaction was completed by TLC, it was cooled, water was added, and dichloromethane was added for extraction. After filtering to remove the insoluble matter, the organic layer was separated, and the water layer was extracted twice with dichloromethane. The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, evaporated to dryness, and a black oil was obtained. Anhydrous ether was added and stirred to solidify, and 0.700 g of brown solid was obtained by suction filtration, i.e., intermediate Dz-3, with a yield of 45%. 1 HNMR (400MHz, DMSO) δ10.32(s,1H),8.51–8.47(m,2H),8.32(dd,J=7.8,2.0Hz,1H),8.19(dd,J=13.5,2.5Hz,1H),7.53(d,J=10.0Hz,2H) ,7.45–7.40(m,2H),6.99(dd,J=7.8,4.8Hz,1H),6.47(dd,J=5.3,1.0Hz,1H),4.39(q,J=7.1Hz,2H),3.95(s,6H),1.37(t,J=7.1Hz,3H).

[0070] 7. Preparation of intermediate mz-4

[0071] mz-3 (0.600 g, 1.35 mmol) was added to 6 mL of tetrahydrofuran, and 6 mL of LiOH (0.064 g, 2.69 mmol) aqueous solution was added under stirring. The reaction was allowed to react overnight at room temperature. After TLC detection, the tetrahydrofuran was concentrated, and a small amount of water was added. The pH was adjusted to 5-6 with dilute hydrochloric acid solution. After stirring for 0.5 hours, 0.360 g of off-white solid was obtained by suction filtration, i.e., intermediate mz-4, with a yield of 64%. 1 H NMR (400MHz, DMSO) δ10.60(s,1H),8.77(d,J=6.7Hz,1H),8.43(s,1H),8.29(d,J=7.7Hz,1H),7.97(s,1H),7.95(s, 1H),7.73(s,1H),7.66(s,1H),7.38(s,1H),7.36(s,1H),6.94(d,J=6.9Hz,1H),6.83(d,J=6.5Hz,1H),4.03(s,6H).

[0072] 8. Preparation of intermediate Dz-4

[0073] Dz-3 (0.700 g, 1.51 mmol) was added to 6 mL of tetrahydrofuran, and 6 mL of LiOH (0.072 g, 3.02 mmol) aqueous solution was added under stirring. The mixture was allowed to react overnight at room temperature. After the reaction was completed as detected by TLC, the tetrahydrofuran was concentrated, and a small amount of water was added. The pH value was adjusted to 5-6 with dilute hydrochloric acid solution. After stirring for 0.5 hour, 0.657 g of off-white solid was obtained by suction filtration, i.e., intermediate Dz-4, with a yield of 77%. 1 H NMR (400MHz, DMSO) δ10.70(s,1H),8.50(d,J=5.3Hz,1H),8.47(dd,J=4.7,2.1Hz,1H),8.30(dd,J=7.7,2.1Hz,1H),8.21(dd,J=13.6 ,2.4Hz,1H),7.56(s,1H),7.53–7.49(m,1H),7.43(d,J=9.0Hz,2H),6.97(dd,J=7.7,4.9Hz,1H),6.49(d,J=5.2Hz,1H),3.96(s,6H).

[0074] Example 2: Preparation of 4 compounds

[0075] 1. Preparation of m-1

[0076] mz-4 (0.100 g, 0.2 mmol), HATU (0.182 g, 0.4 mmol) and DIPEA (0.125 mL, 0.6 mmol) were added to 3 mL of DMF. After stirring for 5 minutes, 1-bicyclo[1,1,1]pentylamine hydrochloride (0.029 g, 0.3 mmol) was added. The mixture was reacted at room temperature for 5 hours. After TLC detection showed that the reaction was complete, water was added. The mixture was stirred for 0.5 hours and then filtered to obtain the crude final product. The light yellow product was separated by preparative thin layer plate, i.e., Example 1. 1 H NMR(400MHz,Chloroform-d)δ10.62(s,1H),8.49(d,J=5.3Hz,1H),8.34(dd,J=4.8,1.7Hz,1H),7.86–7.77(m,2H),7.70(dd,J=7.8,1.7Hz,1H),7.61(s,1 H),7.43(s,1H),7.21–7.11(m,2H),6.72(dd,J=7.7,4.8Hz,1H),6.67(s,1H) ,6.53(d,J=5.3Hz,1H),4.08(s,3H),4.07(s,3H),2.58(s,1H),2.23(s,6H).

[0077] 2. Preparation of m-2

[0078] mz-4 (0.100 g, 0.2 mmol), HATU (0.182 g, 0.4 mmol) and DIPEA (0.125 mL, 0.6 mmol) were added to 3 mL DMF. After stirring for 5 minutes, 3-fluorobicyclo[1,1,1]pentan-1-amine hydrochloride (0.049 g, 0.3 mmol) was added. The mixture was reacted at room temperature for 5 hours. After TLC detection showed that the reaction was complete, water was added. The mixture was stirred for 0.5 hours and then filtered to obtain the crude final product. The light yellow final product was separated by preparative thin layer plate, i.e., Example 2. 1 HNMR (400MHz, CDCl3) δ10.58(s,1H),8.44(d,J=5.3Hz,1H),8.31(dd,J=4.8,1.7Hz,1H),7.79(d,J=1.9Hz,1H),7.77(d,J=4.8Hz,2H),7.58(s,1H) ,7.39(s,1H),7.29(s,1H),7.13(d,J=8.8Hz,2H),6.67(dd,J=7.7,4.8Hz ,1H),6.50(d,J=5.3Hz,1H),4.03(d,J=8.1Hz,6H),2.50(d,J=2.1Hz,6H).

[0079] 3. Preparation of D-1

[0080] Dz-4 (0.100 g, 0.2 mmol), HATU (0.175 g, 0.4 mmol) and DIPEA (0.12 mL, 0.6 mmol) were added to 3 mL DMF. After stirring for 5 minutes, 1-bicyclo[1,1,1]pentylamine hydrochloride (0.041 g, 0.3 mmol) was added. The mixture was reacted at room temperature for 5 hours. After TLC detection showed that the reaction was complete, water was added. The mixture was stirred for 0.5 hours and then filtered to obtain the crude final product. The light yellow final product was separated by preparative thin layer plate, i.e., Example 3. 1H NMR (400MHz, CDCl3) δ10.81(s,1H),8.47(d,J=5.3Hz,1H),8.34(d,J=4.6Hz,1H),8. 05(dt,J=13.2,1.9Hz,1H),7.73(dt,J=7.8,1.6Hz,1H),7.61(s,1H),7.40(s,1H),7 .35(dd,J=8.8,2.4Hz,1H),7.17(t,J=8.8Hz,1H),6.88(s,1H),6.72(dd,J=7.8,4.8 Hz,1H),6.46(d,J=5.3Hz,1H),4.06(s,3H),4.03(s,3H),2.54(s,1H),2.20(s,6H).

[0081] 4. Preparation of D-2

[0082] Dz-4 (0.100 g, 0.2 mmol), HATU (0.175 g, 0.4 mmol) and DIPEA (0.12 mL, 0.6 mmol) were added to 3 mL DMF. After stirring for 5 minutes, 3-fluorobicyclo[1,1,1]pentan-1-amine hydrochloride (0.047 g, 0.3 mmol) was added. The mixture was reacted at room temperature for 5 hours. After TLC detection showed that the reaction was complete, water was added. The mixture was stirred for 0.5 hours and then filtered to obtain the crude final product. The light yellow final product was separated by preparative thin layer plate, i.e., Example 4. 1 HNMR (400MHz, CDCl3) δ10.74(s,1H),8.46(d,J=5.3Hz,1H),8.36(dd,J=5.0,1.7H z,1H),8.06(dd,J=13.0,2.5Hz,1H),7.77(dd,J=7.8,1.8Hz,1H),7.61(s,1H),7.3 9(s,1H),7.38–7.33(m,1H),7.18(t,J=8.8Hz,1H),7.02(s,1H),6.73(dd,J=7.8,4 .7Hz, 1H), 6.46 (d, J = 5.2Hz, 1H), 4.07 (s, 3H), 4.03 (s, 3H), 2.51 (d, J = 2.0Hz, 6H).

[0083] The structural formulas of the four compounds obtained in Example 2 are shown in Table 2.

[0084] Table 2 Chemical names and structural formulas of the four compounds

[0085]

[0086] Example 3 VEGFR-2 inhibitory activity test

[0087] In order to verify whether the prepared 4 compounds have significant inhibitory effects on VEGFR-2, the VEGFR-2 inhibitory activity of the 4 compounds was tested using Lanthascreen assay.

[0088] Experimental Materials:

[0089]

[0090] Experimental steps:

[0091] (1) Prepare kinase buffer, including: 50 mM HEPES (pH 7.5), 10 mM MgCl2, 4 mM DTT, 0.01% Tween-20 and 0.01% BSA.

[0092] (2) Preparation of compound working solutions: Dissolve the four compounds in 100% DMSO and dilute to 100 times the maximum concentration required in the reaction, i.e., 400 μM. Then, further dilute them 4-fold to 9 concentrations using 100% DMSO, for a total of 10 concentrations, i.e., 400 μM, 100 μM, 25 μM, 6.25 μM, 1.5625 μM, 0.3906 μM, 0.0976 μM, 0.0244 μM, 0.0061 μM, and 0.0015 μM.

[0093] (3) Preparation of VEGFR-2 kinase solution: VEGFR-2 kinase was prepared into a kinase solution with a concentration of 0.1 nM using a kinase buffer.

[0094] (4) Preparation of substrate solution: Fluorescein-Poly GT and ATP were prepared into a substrate solution with a concentration of 0.04 uM Fluorescein-Poly GT and 200 uM ATP using kinase buffer.

[0095] (5) Preparation of detection solution: Use detection antibody buffer to prepare the concentration to 2 nM Tb-PY20Antibody and 40mM EDTA detection solution.

[0096] (6) Kinase reaction: 200 nL of the four compounds (10 concentrations) were transferred to a 384-well test plate in triplicate. Then, 10 μL of VEGFR-2 kinase solution was transferred to the 384-well test plate. A control group without compound (with DMSO instead) and a control group without enzyme (with buffer instead) were set up separately. After incubation at room temperature for 10 minutes, 10 μL of substrate solution was transferred to the 384-well test plate. The reaction started, and at this time, the final concentrations of the example compounds were 4000 nM, 1000 nM, 250 nM, 62.50 nM, 15.63 nM, 3.906 nM, 0.977 nM, 0.2441 nM, 0.0610 nM, and 0.01526 nM. After incubation at room temperature for 30 minutes, 20 μL of detection solution was added to terminate the reaction, centrifuged for 30 seconds to mix, and incubated at room temperature for 60 minutes.

[0097] (7) Data reading

[0098] Fluorescence intensity (RFU) was collected on Envision at an excitation wavelength of 340 nm and emission wavelengths of 520 nm and 495 nm.

[0099] (8) Data processing

[0100] Calculate the ratio of RFU 520nm / RFU 495nm. Convert the ratio to percent inhibition rate (%), inhibition rate (%) = (RFU 520nm / RFU 495nm) max -RFU compound ) / (RFU max -RFU min )*100. min ” represents the ratio of the no enzyme control group, “RFU max " indicates the ratio of the control group without compound addition. According to the inhibition rate, Graphpadprism software was used for curve fitting to calculate the IC of each example compound. 50 value.

[0101] Table 3 Inhibitory activity of four compounds on VEGFR-2

[0102] Example <![CDATA[IC 50 (nM)]]> m-1 24.75 m-2 22.51 D-1 20.50 D-2 27.56

[0103] in conclusion

[0104] From Table 3 and Figure 2 It can be seen that the four compounds all showed outstanding inhibitory activity against VEGFR-2, IC 50 The values ​​were all below 30 nM.

[0105] In summary, the example compounds disclosed in the present invention can show significant inhibitory activity against VEGFR-2 and have the potential to treat VEGFR-2-mediated related diseases.

[0106] It should be understood that the present invention is not limited to the specific embodiments of the invention described above, as variations may be made thereto and still fall within the scope of the appended claims.

Claims

1. A compound represented by the general structural formula I, its optical isomers, isotopic derivatives or pharmaceutically acceptable salts thereof, in, R1 is selected from H, halogen, C 1-3 Alkyl, C 1-3 Alkoxy or C 1-3 Alkylamino; R2 is selected from H, halogen, C 1-3 Alkyl, C 1-3 Alkoxy or C 1-3 Alkylthio; R3 is selected from H or halogen; X is selected from C or N.

2. The compound of formula I as claimed in claim 1, its optical isomers, isotopic derivatives or pharmaceutically acceptable salts thereof, wherein R1 is selected from H or halogen.

3. The compound of formula I according to claim 1, its optical isomers, isotopic derivatives or pharmaceutically acceptable salts thereof, wherein R2 is selected from H, halogen or C 1-3 alkyl.

4. The compound of formula I as claimed in claim 1, its optical isomers, isotopic derivatives or pharmaceutically acceptable salts thereof, wherein X is C.

5. The compound of formula I according to any one of claims 1 to 4, its optical isomers, isotopic derivatives or Its pharmaceutically acceptable salt, the compound is selected from: 、 、 、 。 6. A pharmaceutical composition comprising the compound according to any one of claims 1 to 5, its optical isomer, isotopic derivative or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable excipients or carriers.

7. Use of the compound according to any one of claims 1 to 5, its optical isomer, isotopic derivative or pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 6 in the preparation of a VEGFR-2 inhibitor.

8. Use of the compound according to any one of claims 1 to 5, its optical isomer, isotopic derivative or pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 6 in the preparation of a medicament for treating VEGFR-2 kinase-mediated related diseases.

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