Synthesis method of novel estrogen receptor targeting inhibitor and application of novel estrogen receptor targeting inhibitor in breast cancer treatment
阅读说明：本技术 新型雌激素受体靶向性抑制剂合成方法及在乳腺癌治疗中的应用 (Synthesis method of novel estrogen receptor targeting inhibitor and application of novel estrogen receptor targeting inhibitor in breast cancer treatment ) 是由 杨锦飞 姚伟伟 于 2021-07-12 设计创作，主要内容包括：本发明提供了一种新型雌激素受体靶向性抑制剂合成方法及在乳腺癌治疗中的应用,属于医药化工应用技术领域。其技术方案为：通过合成了一系列含有吲唑和吲哚骨架的雌激素受体靶向性抑制剂,并且分别测试了它们对乳腺癌细胞的抑制活性,发现了一系列比氯尼达明具有更高癌细胞抑制活性的化合物；其中,化合物(21)和(23)显示出最好的乳腺癌细胞抑制活性,IC-(50)值分别为45μM和53μM。本发明的有益效果为：本发明提供了一系列比氯尼达明,具有更高乳腺癌癌细胞抑制活性的小分子化合物,其中,化合物(21)和(23)显示出最强的生物活性,分子对接实验结果证明它们是通过靶向雌激素受体发挥其对乳腺癌细胞的抑制作用。(The invention provides a synthesis method of a novel estrogen receptor targeting inhibitor and application of the novel estrogen receptor targeting inhibitor in breast cancer treatment, belonging to the technical field of pharmaceutical and chemical application. The technical scheme is as follows: a series of estrogen receptor targeted inhibitors containing indazole and indole frameworks are synthesized, and the inhibitory activity of the inhibitors on breast cancer cells is tested respectively, so that a series of compounds with higher cancer cell inhibitory activity than lonidamine are discovered; among them, the compounds (21) and (23) showed the best breast cancer cell inhibitory activity, IC 50 The values were 45. mu.M and 53. mu.M, respectively. The invention has the beneficial effects that: the invention provides a series of micromolecular compounds with higher breast cancer cell inhibition activity than lonidamine, wherein the compounds (21) and (23) show the strongest biological activityThe results of molecular docking experiments prove that the compounds exert the inhibition effect on breast cancer cells by targeting estrogen receptors.)
1. A method for synthesizing a novel estrogen receptor targeting inhibitor is characterized in that,
the synthetic route of the compound (4) is as follows:
the specific content of the synthesis method of the compound (4) comprises the following steps:
firstly, indazole formate (1) and m-tri-substituted benzyl chloride (2) are used as raw materials, alkali in reaction is one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate or dipotassium hydrogen phosphate, a solvent is one of acetone, cyclohexanone, acetonitrile, tetrahydrofuran or 1, 4-epoxy hexacyclic ring, the reaction is carried out in the air, the reaction temperature is 60-80 ℃, and the reaction time is 6-24 hours;
secondly, the alkali used in the reduction reaction of the compound (3) is one of sodium hydroxide and potassium hydroxide, the solvent is one of methanol/water, ethanol/water, tetrahydrofuran/water or acetone/water, the temperature is any temperature between room temperature and 60 ℃, and the reaction time is any time between 12 and 36 hours;
the synthetic route of the compound (5) is as follows:
the specific content of the synthesis method of the compound (5) comprises the following steps:
firstly, indazole formate (1) and m-tri-substituted benzyl chloride (2) are used as raw materials, alkali in reaction is one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate or dipotassium hydrogen phosphate, a solvent is one of acetone, cyclohexanone, acetonitrile, tetrahydrofuran or 1, 4-epoxy hexacyclic ring, the reaction is carried out in the air, the reaction temperature is 60-80 ℃, and the reaction time is 6-24 hours;
secondly, hydrazine hydrate is used as a hydrazinolysis reagent, one of ethanol, methanol or isopropanol is used as a solvent, the temperature is any temperature between 60 ℃ and 120 ℃, and the reaction time is any time between 2 hours and 6 hours;
the synthetic route of the compound (8) is as follows:
the specific content of the synthesis method of the compound (8) comprises the following steps:
firstly, 2H-indazole-5-carboxylic acid methyl ester (6) and 2, 4-dichloro-1- (chloromethyl) benzene (2a) are used as raw materials, alkali in reaction is one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate or dipotassium hydrogen phosphate, a solvent is one of acetone, cyclohexanone, acetonitrile, tetrahydrofuran or 1, 4-epoxy hexacyclic ring, the reaction is carried out in air, the reaction temperature is 60-80 ℃, and the reaction time is 6-24 hours;
secondly, hydrazine hydrate is used as a hydrazinolysis reagent, one of ethanol, methanol and isopropanol is used as a solvent, the temperature is any temperature between 60 ℃ and 120 ℃, and the reaction time is any time between 2 hours and 6 hours;
the synthetic route of the compound (9) is as follows:
the specific content of the synthesis method of the compound (9) comprises the following steps:
the method comprises the following steps of firstly, taking 1- (2, 4-dichlorobenzyl) -1H-indazole-3-carboxylic acid (4a) as a raw material, taking a chlorination reagent in a reaction as one of thionyl chloride and oxalyl chloride, taking a solvent as one of tetrahydrofuran, 1, 4-dioxane, acetonitrile and acetone, carrying out the reaction in an inert gas at the reaction temperature of between room temperature and 50 ℃ for 2 hours;
secondly, using ammonia water as an amination reagent, and reacting at room temperature for any time between 10 and 30 minutes;
the synthetic route of the synthetic method of the compound (13) is as follows:
the specific content of the synthesis method of the compound (13) comprises the following steps:
the first step, 1H-indole-3-carboxylic acid methyl ester (10) and 2, 4-dichloro-1- (chloromethyl) benzene (2a) are used as raw materials, alkali in the reaction is one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate or dipotassium hydrogen phosphate, a solvent is one of acetone, cyclohexanone, acetonitrile, tetrahydrofuran or 1, 4-epoxy six-ring, the reaction is carried out in the air, the reaction temperature is 60-80 ℃, and the reaction time is 6-24 hours;
in the second step of reaction, hydrazine hydrate is used as a hydrazinolysis reagent, one of ethanol, methanol and isopropanol is used as a solvent, the temperature is any temperature between 60 ℃ and 120 ℃, and the reaction time is any time between 2 hours and 6 hours;
the synthetic route of the synthetic method of the compound (14) is as follows:
the specific content of the synthesis method of the compound (14) comprises the following steps:
the first step, 1H-indole-3-carboxylic acid methyl ester (10) and 2, 4-dichloro-1- (chloromethyl) benzene (2a) are used as raw materials, alkali in the reaction is one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate or dipotassium hydrogen phosphate, a solvent is one of acetone, cyclohexanone, acetonitrile, tetrahydrofuran or 1, 4-epoxy six-ring, the reaction is carried out in the air, the reaction temperature is 60-80 ℃, and the reaction time is 6-24 hours;
secondly, the alkali used in the reduction reaction of the compound (11) is one of sodium hydroxide and potassium hydroxide, the solvent is one of methanol/water, ethanol/water, tetrahydrofuran/water and acetone/water, the temperature is any temperature between room temperature and 60 ℃, and the reaction time is any time between 12 and 36 hours;
thirdly, in the reaction of the compound (12), a chlorination reagent is one of thionyl chloride and oxalyl chloride, a solvent is one of tetrahydrofuran, 1, 4-dioxane, acetonitrile or acetone, the reaction is carried out in inert gas, the reaction temperature is between room temperature and 50 ℃, and the reaction time is 2 hours;
fourthly, the amination reagent is ammonia water, the temperature is room temperature, and the reaction time is any time between 10 and 30 minutes;
the synthetic route of the synthetic method of the compound (19) is as follows:
the specific content of the synthesis method of the compound (19) comprises the following steps:
step one, indazole (15) is used as a raw material, alkali in reaction is one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate or dipotassium hydrogen phosphate, an iodinating reagent is iodine, a reaction solvent is N, N-dimethylformamide, and the reaction is carried out in air at room temperature;
secondly, the alkali used in the coupling reaction of the compound (16) and the 2, 4-dichloro-1- (chloromethyl) benzene (2a) is one of cesium carbonate, potassium carbonate, sodium carbonate and silver carbonate, the solvent is one of acetone, cyclohexanone, acetonitrile, tetrahydrofuran or 1, 4-epoxy hexacyclic ring, the temperature is any one of 60-80 ℃, and the reaction time is any one of 6-24 hours;
thirdly, a catalyst used in the coupling reaction of the compound (17) and allyl formate is one of palladium acetate, palladium chloride, palladium bromide and palladium trifluoroacetate, a base is one of N, N-diisopropylethylamine and triethylamine, a phosphine ligand is one of triphenylphosphine, tricyclohexylphosphine and 1, 2-bis (diphenylphosphino) ethane, a reaction solvent is N, N-dimethylformamide, the temperature in the reaction is any one of 100-120 ℃, and the reaction time is any time between 12 and 24 hours;
fourthly, hydrazine hydrate is used as a hydrazinolysis reagent, one of ethanol, methanol and isopropanol is used as a solvent, the temperature is any temperature between 60 ℃ and 120 ℃, and the reaction time is any time between 2 hours and 6 hours;
the synthetic route of the synthetic method of the compound (21) is as follows:
the specific content of the synthesis method of the compound (21) comprises the following steps:
firstly, taking a compound (17) and (4- (methoxycarbonyl) phenyl) boric acid as raw materials, taking a metal catalyst in the reaction, namely one of palladium acetate, palladium chloride, palladium bromide or palladium trifluoroacetate, taking alkali as one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate or dipotassium hydrogen phosphate, taking a reaction solvent as a mixed solvent of ethanol/toluene/water, carrying out the reaction in an inert gas at any temperature of 80-120 ℃ for any time of 12-36 hours;
in the second step of reaction, hydrazine hydrate is used as a hydrazinolysis reagent, one of ethanol, methanol and isopropanol is used as a solvent, the temperature is any temperature between 60 ℃ and 120 ℃, and the reaction time is any time between 2 hours and 6 hours;
the synthetic route of the compound (23) is as follows:
the specific content of the synthesis method of the compound (23) comprises the following steps:
firstly, taking a compound (17) and (3- (methoxycarbonyl) phenyl) boric acid as raw materials, taking a metal catalyst in the reaction, namely one of palladium acetate, palladium chloride, palladium bromide or palladium trifluoroacetate, taking alkali as one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate or dipotassium hydrogen phosphate, taking a reaction solvent as a mixed solvent of ethanol/toluene/water, carrying out the reaction in an inert gas at any temperature of 80-120 ℃ for any time of 12-36 hours;
and in the second step, the hydrazinolysis reagent is hydrazine hydrate, the solvent is one of ethanol, methanol and isopropanol, the temperature is any temperature between 60 ℃ and 120 ℃, and the reaction time is any time between 2 hours and 6 hours.
2. The novel estrogen receptor targeted inhibitor synthesis method according to claim 1, wherein, in the first step of the synthesis method of the compound (4),
the alkali is one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate or dipotassium hydrogen phosphate, and the dosage range of the alkali is as follows: 200 and 500mol percent;
the ratio of the dosage of the alkali to the dosage of the compound (1) is as follows: 2:1-5:1, the amount of the base substance is 2-5 times of the amount of the compound (1) substance;
the position of the methyl formate in the compound (1) may be at any one of positions 3, 4, 5, 6 and 7 of the indazole;
the position of the methyl formate in the compound (3) may be at any one of positions 3, 4, 5, 6 and 7 of the indazole;
the position of the formic acid in the compound (4) may be at any one of the 3-, 4-, 5-, 6-and 7-positions of the indazole;
r in the compound (2)1Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group;
R2is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group;
R3is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group;
r in the compound (3)1Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group;
R2is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group;
R3is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group;
r in the compound (4)1Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group;
R2is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group;
R3is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group;
the molar part ratio range of the compound (1) to the compound (2) is as follows: 1:2-1: 1;
the solvent comprises acetone, cyclohexanone, acetonitrile, tetrahydrofuran and 1, 4-epoxy hexacyclic ring;
the reaction temperature is 60-80 ℃;
the reaction time is any time within 6 to 24 hours;
in the second reaction step in the synthesis method of the compound (4),
the alkali is one of sodium hydroxide and potassium hydroxide, and the dosage range of the alkali is as follows: 100-200 mol%;
the ratio of the used amount of the alkali to the used amount of the compound (3) is as follows: 1:1-2:1, the amount of the base substance being 1-2 times the amount of the compound (1) substance;
the solvent comprises methanol/water, ethanol/water, tetrahydrofuran/water and acetone/water;
the reaction temperature is any temperature between room temperature and 60 ℃;
the reaction time is any time between 12 and 36 hours.
3. The novel estrogen receptor targeted inhibitor synthesis method according to claim 1, wherein, in the first step of the compound (5) synthesis method,
the alkali is one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate and dipotassium hydrogen phosphate, and the dosage range of the alkali is as follows: 200 and 500mol percent;
the position of the methyl formate in the compound (1) may be at any one of positions 3, 4, 5, 6 and 7 of the indazole;
the position of the methyl formate in the compound (3) may be at any one of positions 3, 4, 5, 6 and 7 of the indazole;
the position of the hydrazide in the compound (5) may be at any one of positions 3, 4, 5, 6 and 7 of the indazole;
r in the compound (2)1Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group; r2Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group; r3Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group;
r in the compound (3)1Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group; r2Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group; r3Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group;
r in the Compound (5)1Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group; r2Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group; r3Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group;
the ratio of the dosage of the alkali to the dosage of the compound (1) is as follows: 2:1-5:1, the amount of the base substance is 2-5 times of the amount of the compound (1) substance;
the molar part ratio range of the compound (1) to the compound (2) is as follows: 1:2-1: 1;
the solvent comprises acetone, cyclohexanone, acetonitrile, tetrahydrofuran and 1, 4-epoxy hexacyclic ring;
the reaction temperature is 60-80 ℃;
the reaction time is any time within 6 to 24 hours;
in the second reaction step in the synthesis method of the compound (5),
the alkali is hydrazine hydrate, and the dosage range of the hydrazine hydrate is as follows: 100-200 mol%;
the dosage of the hydrazine hydrate and the material ratio of the compound (3) are as follows: 1:1-2: 1;
the solvent comprises methanol, ethanol and isopropanol;
the reaction temperature is any temperature between 60 ℃ and 120 ℃;
the reaction time is any time between 2 and 6 hours.
4. The novel estrogen receptor targeted inhibitor synthesis method according to claim 1, characterized in that, in the first step of the compound (8) synthesis method,
the alkali is one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate and dipotassium hydrogen phosphate, and the dosage range of the alkali is as follows: 200 and 500mol percent;
the ratio of the dosage of the alkali to the dosage of the compound (1) is as follows: 2:1-5:1, the amount of the base substance is 2-5 times of the amount of the compound (1) substance;
the molar part ratio range of the compound (1) to the compound (2) is as follows: 1:2-1: 1;
the solvent comprises acetone, cyclohexanone, acetonitrile, tetrahydrofuran and 1, 4-epoxy hexacyclic ring;
the reaction temperature is 60-80 ℃;
the reaction time is any time within 6 to 24 hours;
in the second reaction step in the synthesis method of the compound (8),
the alkali is hydrazine hydrate, and the dosage range of the hydrazine hydrate is as follows: 100-200 mol%;
the dosage of the hydrazine hydrate and the dosage ratio of the compound (7) are as follows: 1:1-2: 1;
the solvent comprises methanol, ethanol and isopropanol;
the reaction temperature is any temperature between 60 ℃ and 120 ℃;
the reaction time is any time between 2 and 6 hours.
5. The novel estrogen receptor targeted inhibitor synthesis method according to claim 1, characterized in that, in the first step of the compound (9) synthesis method,
the chlorination reagent is one of thionyl chloride and oxalyl chloride, and the dosage range of the chlorination reagent is as follows: 110-200 mol%;
the dosage of the chlorinating agent and the dosage ratio of the compound (4a) are as follows: 1.1:1-2:1, the amount of the substance of the chlorinating reagent is 1.1-2 times of the amount of the substance of the compound (1);
the solvent comprises tetrahydrofuran, 1, 4-dioxane, acetonitrile and acetone;
reacting in the inert gas atmosphere, wherein the inert gas comprises nitrogen, helium and argon;
the reaction temperature is any one of room temperature to 50 ℃;
the reaction time is any time within 2 to 6 hours;
in the second reaction step in the synthesis method of the compound (9),
the amination reagent is ammonia water, and the dosage range of the ammonia water is as follows: 100-200 mol%;
the dosage of the hydrazine hydrate and the dosage ratio of the compound (4a) are as follows: 1:1-2: 1;
the reaction temperature is room temperature;
the reaction time is any time between 10 and 30 minutes.
6. The novel estrogen receptor targeted inhibitor synthesis method according to claim 1, wherein, in the first step of the compound (13) synthesis method,
the alkali is one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate or dipotassium hydrogen phosphate, and the dosage range of the alkali is as follows: 200 and 500mol percent;
the ratio of the dosage of the alkali to the dosage of the compound (10) is as follows: 2:1-5:1, the amount of the base substance is 2-5 times of the amount of the compound (1) substance;
the molar part ratio of the compound (110) to the compound (2a) is as follows: 1:2-1: 1;
the solvent comprises acetone, cyclohexanone, acetonitrile, tetrahydrofuran and 1, 4-epoxy hexacyclic ring;
the reaction temperature is 60-80 ℃;
the reaction time is any time within 6 to 24 hours;
in the second reaction step in the synthesis method of the compound (13),
the alkali is hydrazine hydrate, and the dosage range of the hydrazine hydrate is as follows: 100-200 mol%;
the dosage of the hydrazine hydrate and the dosage ratio of the compound (11) are as follows: 1:1-2: 1;
the solvent comprises methanol, ethanol and isopropanol;
the reaction temperature is any temperature between 60 ℃ and 120 ℃;
the reaction time is any time between 2 and 6 hours.
7. The novel estrogen receptor targeted inhibitor synthesis method according to claim 1, wherein, in the first step of the compound (14) synthesis method,
the alkali is one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate and dipotassium hydrogen phosphate, and the dosage range of the alkali is as follows: 200 and 500mol percent;
the ratio of the dosage of the alkali to the dosage of the compound (10) is as follows: 2:1-5:1, the amount of the base substance is 2-5 times of the amount of the compound (1) substance;
the molar part ratio of the compound (110) to the compound (2a) is as follows: 1:2-1: 1;
the solvent comprises acetone, cyclohexanone, acetonitrile, tetrahydrofuran and 1, 4-epoxy hexacyclic ring;
the reaction temperature is 60-80 ℃;
the reaction time is any time within 6 to 24 hours;
in the second reaction step in the synthesis method of the compound (14),
the alkali is one of sodium hydroxide and potassium hydroxide, and the dosage range of the alkali is as follows: 100-200 mol%;
the ratio of the dosage of the alkali to the dosage of the compound (1) is as follows: 1:1-2:1, the amount of the base substance being 1-2 times the amount of the compound (1) substance;
the solvent comprises methanol/water, ethanol/water, tetrahydrofuran/water and acetone/water;
the reaction temperature is any temperature between room temperature and 60 ℃;
the temperature is any temperature between room temperature and 60 ℃;
the reaction time is any time between 12 and 36 hours;
in the third reaction step in the synthesis method of the compound (14),
the chlorination reagent is one of thionyl chloride and oxalyl chloride, and the dosage range of the chlorination reagent is as follows: 110-200 mol%;
the dosage of the chlorinating agent and the dosage ratio of the compound (4a) are as follows: 1.1:1-2:1, the amount of the substance of the chlorinating reagent is 1.1-2 times of the amount of the substance of the compound (1);
the solvent comprises tetrahydrofuran, 1, 4-dioxane, acetonitrile and acetone;
reacting in the inert gas atmosphere, wherein the inert gas comprises nitrogen, helium and argon;
the reaction temperature is any one of room temperature to 50 ℃;
the reaction time is any time within 2 to 6 hours;
in the fourth reaction step in the synthesis method of the compound (14),
the amination reagent is ammonia water, and the dosage range of the ammonia water is as follows: 100-200 mol%;
the ratio of the dosage of the ammonia water to the dosage of the compound (12) is as follows: 1:1-2: 1;
the reaction temperature is room temperature;
the reaction time is any time between 10 and 30 minutes.
8. The novel estrogen receptor targeted inhibitor synthesis method according to claim 1, wherein, in the first step of the synthesis method of the compound (19),
the alkali is one of sodium hydroxide and potassium hydroxide, and the dosage range of the alkali is as follows: 100-200 mol%;
the ratio of the used amount of the alkali to the used amount of the compound (15) is as follows: 1:1-2:1, the amount of the base substance being 1-2 times the amount of the compound (1) substance;
the compound (15) is reacted with iodine I2The molar ratio ranges are as follows: 1:2-1: 1;
the solvent is N, N-dimethylformamide;
the reaction temperature is room temperature;
the reaction time is any time within 2 to 4 hours;
in the second reaction step in the synthesis method of the compound (19),
the alkali is one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate and dipotassium hydrogen phosphate, and the dosage range of the alkali is as follows: 200 and 500mol percent;
the ratio of the dosage of the alkali to the dosage of the compound (10) is as follows: 2:1-5:1, the amount of the base substance is 2-5 times of the amount of the compound (1) substance;
the molar part ratio of the compound (110) to the compound (2a) is as follows: 1:2-1: 1;
the solvent comprises acetone, cyclohexanone, acetonitrile, tetrahydrofuran and 1, 4-epoxy hexacyclic ring;
the reaction temperature is room temperature;
the reaction time is any time within 6 to 24 hours;
in the third reaction step in the synthesis method of the compound (19),
the metal palladium catalyst is one of palladium acetate, palladium chloride, palladium bromide and palladium trifluoroacetate, and the dosage range of the catalyst is as follows: 10-20 mol%;
the phosphine ligand is one of triphenylphosphine, tricyclohexylphosphine and 1, 2-bis (diphenylphosphino) ethane, and the dosage range of the phosphine ligand is as follows: 25-50 mol%;
the alkali is one of N, N-diisopropylethylamine and triethylamine, and the dosage range of the alkali is as follows: 200-300 mol%;
the molar part ratio of the compound (17) to the allyl formate is as follows: 1:2-1: 5;
the reaction solvent is N, N-dimethylformamide;
reacting in the inert gas atmosphere, wherein the inert gas comprises nitrogen, helium and argon;
the temperature in the reaction is any one of 100 ℃ to 120 ℃,
the reaction time is any time between 12 and 24 hours;
in the fourth reaction step in the synthesis method of the compound (19),
the alkali is hydrazine hydrate, and the dosage range of the hydrazine hydrate is as follows: 100-200 mol%;
the dosage of the hydrazine hydrate and the dosage ratio of the compound (18) are as follows: 1:1-2: 1;
the solvent comprises methanol, ethanol and isopropanol;
the reaction temperature is any temperature between 60 ℃ and 120 ℃;
the reaction time is any time between 2 and 6 hours;
in the first reaction step in the synthesis method of the compound (21),
the metal palladium catalyst is one of palladium acetate, palladium chloride, palladium bromide and palladium trifluoroacetate, and the dosage range of the catalyst is as follows: 10-20 mol%;
the alkali is one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate and dipotassium hydrogen phosphate, and the dosage range of the alkali is as follows: 100-200 mol%;
the ratio of the used amount of the alkali to the used amount of the compound (17) is as follows: 1:1-2:1, the amount of the base substance being 1-2 times the amount of the compound (1) substance;
the molar ratio of the compound (17) to the (4- (methoxycarbonyl) phenyl) boric acid is as follows: 1:2-1: 1;
the solvent is a mixed solvent of ethanol/toluene/water;
reacting in the inert gas atmosphere, wherein the inert gas comprises nitrogen, helium and argon;
the reaction temperature is any one of 60 ℃ to 80 ℃;
the reaction time is any time within 12 hours to 24 hours;
in the second reaction step in the synthesis method of the compound (21),
the alkali is hydrazine hydrate, and the dosage range of the hydrazine hydrate is as follows: 100-200 mol%;
the dosage of the hydrazine hydrate and the dosage ratio of the compound (20) are as follows: 1:1-2: 1;
the solvent comprises methanol, ethanol and isopropanol;
the reaction temperature is any temperature between 60 ℃ and 120 ℃;
the reaction time is any time between 2 and 6 hours;
in the first reaction step in the synthesis method of the compound (23),
the metal palladium catalyst is one of palladium acetate, palladium chloride, palladium bromide and palladium trifluoroacetate, and the dosage range of the catalyst is as follows: 10-20 mol%;
the alkali is one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate and dipotassium hydrogen phosphate, and the dosage range of the alkali is as follows: 100-200 mol%;
the ratio of the used amount of the alkali to the used amount of the compound (17) is as follows: 1:1-2:1, the amount of the base substance being 1-2 times the amount of the compound (1) substance;
the molar ratio of the compound (17) to the (3- (methoxycarbonyl) phenyl) boric acid is as follows: 1:2-1: 1;
the solvent is a mixed solvent of ethanol/toluene/water;
reacting in the inert gas atmosphere, wherein the inert gas comprises nitrogen, helium and argon;
the reaction temperature is any one of 60 ℃ to 80 ℃;
the reaction time is any time within 12 hours to 24 hours;
in the second reaction step in the synthesis method of the compound (23),
the alkali is hydrazine hydrate, and the dosage range of the hydrazine hydrate is as follows: 100-200 mol%;
the ratio of the usage amount of the hydrazine hydrate to the usage amount of the compound (22) is as follows: 1:1-2:1,
the solvent comprises methanol, ethanol and isopropanol;
the reaction temperature is any temperature between 60 ℃ and 120 ℃;
the reaction time is any time between 2 and 6 hours;
the synthesis method of the compound (4) comprises the following specific steps:
the first step of reaction: sequentially adding weighed indazole formate (1), alkali and m-tri-substituted benzyl chloride (2) into a reaction bottle, adding a solvent, reacting at 60-80 ℃, stirring for 6-24 hours, adding water to quench the reaction after the reaction is finished, then adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, and performing column chromatography separation and purification to obtain a target compound (3);
the second step of reaction: sequentially adding the weighed compound 3, alkali and solvent into a reaction bottle, reacting in an inert gas atmosphere at room temperature-60 ℃, stirring for 12-36 hours, adding water to quench the reaction after the reaction is finished, then adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, and performing column chromatography separation and purification to obtain a target compound (4);
the synthesis method of the compound (5) comprises the following specific steps:
the first step of reaction: sequentially adding weighed indazole formate (1), alkali and m-tri-substituted benzyl chloride (2) into a reaction bottle, adding a solvent, reacting at 60-80 ℃, stirring for 6-24 hours, adding water to quench the reaction after the reaction is finished, then adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, and performing column chromatography separation and purification to obtain a target compound (3);
the second step of reaction: sequentially adding the weighed compound (3), hydrazine hydrate and solvent into a reaction bottle, then carrying out inert gas protection, reacting in an inert gas atmosphere, stirring at 60-120 ℃ for 2-6 hours, adding water to quench the reaction after the reaction is finished, then adding ethyl acetate, separating an organic phase by using a separating funnel, combining the organic phases, carrying out spin drying, and carrying out column chromatography separation and purification to obtain a target compound (5);
the synthesis method of the compound (8) comprises the following specific steps:
the first step of reaction: sequentially adding weighed 2H-indazole-5-carboxylic acid methyl ester (6), alkali and 2, 4-dichloro-1- (chloromethyl) benzene (2a) into a reaction bottle, reacting at 60-80 ℃, stirring for 6-24 hours, adding water to quench the reaction after the reaction is finished, then adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, drying by spinning, and performing column chromatography separation and purification to obtain a target compound (7);
the second step of reaction: sequentially adding the weighed compound (4), hydrazine hydrate and solvent into a reaction bottle, reacting in an inert gas atmosphere at 60-120 ℃ for 2-6 hours, adding water to quench the reaction after the reaction is finished, adding ethyl acetate, filtering the solid by using a suction filter funnel, separating an organic phase by using a separating funnel, combining the organic phases, drying by spinning, and performing column chromatography separation and purification to obtain a target compound (8);
the synthesis method of the compound (9) comprises the following specific steps:
the first step of reaction: sequentially adding weighed 1- (2, 4-dichlorobenzyl) -1H-indazole-3-carboxylic acid (4a) and a solvent into a reaction bottle, then carrying out inert gas protection, slowly dropwise adding a chlorination reagent at 0 ℃, reacting in an inert gas atmosphere, stirring for 2 hours at 50 ℃, adding water to quench the reaction after the reaction is finished, then adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, and directly carrying out the next reaction after spin-drying;
the second step of reaction: sequentially adding the crude product and ammonia water in the last step into a reaction bottle, reacting in an inert gas atmosphere, stirring at room temperature for 10-30 minutes, adding water to quench the reaction after the reaction is finished, then adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, performing column chromatography separation and purification after spin-drying to obtain a target compound (9);
the synthesis method of the compound (13) comprises the following specific steps:
the first step of reaction: sequentially adding weighed 1H-indole-3-carboxylic acid methyl ester (10), alkali and 2, 4-dichloro-1- (chloromethyl) benzene (2a) into a reaction bottle, adding a reaction solvent, reacting, stirring for 6-24 hours at 60-80 ℃, adding water for quenching reaction after the reaction is finished, then adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, and performing column chromatography separation and purification after spin drying to obtain a target compound (11);
the second step of reaction: sequentially adding the weighed compound (11), hydrazine hydrate and solvent into a reaction bottle, then carrying out inert gas protection, stirring for 2-6 hours at the temperature of 60-120 ℃ for reaction, adding water to quench the reaction after the reaction is finished, then adding ethyl acetate, separating an organic phase by using a separating funnel, combining the organic phases, carrying out spin drying, and carrying out column chromatography separation and purification to obtain a target compound (13);
the synthesis method of the compound (14) comprises the following specific steps:
the first step of reaction: sequentially adding weighed 1H-indole-3-carboxylic acid methyl ester (10), alkali and 2, 4-dichloro-1- (chloromethyl) benzene (2a) into a reaction bottle, then adding a reaction solvent, reacting, stirring for 6-24 hours at 60-80 ℃, adding water for quenching reaction after the reaction is finished, then adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, and performing column chromatography separation and purification after spin drying to obtain a target compound (11);
the second step of reaction: sequentially adding the weighed compound (11), alkali and solvent into a reaction bottle, reacting in an inert gas atmosphere at room temperature of 60 ℃ below zero, stirring for 12-36 hours, adding water to quench the reaction after the reaction is finished, adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, drying by spinning, and performing column chromatography separation and purification to obtain a target compound (12);
the third step of reaction: sequentially adding the weighed compound (12) and a solvent into a reaction bottle, then carrying out inert gas protection, slowly dropwise adding a chlorination reagent at 0 ℃, reacting in an inert gas atmosphere, stirring for 2 hours at 50 ℃, adding water to quench the reaction after the reaction is finished, then adding ethyl acetate, separating an organic phase by using a separating funnel, combining the organic phases, and directly carrying out the next reaction after spin-drying;
and a fourth step of reaction: sequentially adding the crude product and ammonia water in the last step into a reaction bottle, reacting in an inert gas atmosphere, stirring at room temperature for 10-30 minutes, adding water to quench the reaction after the reaction is finished, then adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, performing column chromatography separation and purification after spin-drying to obtain a target compound (14);
the synthesis method of the compound (19) comprises the following specific steps:
the first step of reaction: adding weighed indazole (15), iodination reagent and solvent into a reaction bottle in sequence, reacting, stirring at room temperature for 1-3 hours, adding water to quench the reaction after the reaction is finished, adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, performing column chromatography separation and purification after spin drying to obtain a target compound (16);
the second step of reaction: sequentially adding the weighed compound (16), alkali and 2, 4-dichloro-1- (chloromethyl) benzene (2a) into a reaction bottle, then adding a reaction solvent, reacting, stirring for 6-24 hours at the temperature of 60-80 ℃, adding water to quench the reaction after the reaction is finished, then adding ethyl acetate, separating an organic phase by using a separating funnel, combining the organic phases, carrying out spin drying, and carrying out column chromatography separation and purification to obtain a target compound (17);
the third step of reaction: sequentially adding the weighed compound (17), a palladium catalyst, alkali, a phosphine ligand and allyl formate into a reaction bottle, then adding a reaction solvent, then carrying out inert gas protection, stirring for 12-24 hours at the temperature of 100-120 ℃ for reaction, adding water to quench the reaction after the reaction is finished, then adding ethyl acetate, filtering the solid by using a suction filter funnel, then separating the organic phase by using a separating funnel, combining the organic phases, carrying out spin drying, and carrying out column chromatography separation and purification to obtain a target compound (18);
and a fourth step of reaction: and (2) sequentially adding the weighed compound (18), hydrazine hydrate and solvent into a reaction bottle, then carrying out inert gas protection, stirring for 2-6 hours at the temperature of 60-120 ℃ for reaction, adding water to quench the reaction after the reaction is finished, then adding ethyl acetate, separating an organic phase by using a separating funnel, combining the organic phases, carrying out spin drying, and carrying out column chromatography separation and purification to obtain the target compound (19).
9. The method for synthesizing the novel estrogen receptor targeted inhibitor according to claim 1, wherein the specific steps in the method for synthesizing the compound (21) are as follows:
the first step of reaction: sequentially adding the weighed compound (17), a palladium catalyst, alkali and (4- (methoxycarbonyl) phenyl) boric acid into a reaction bottle, then adding a reaction solvent, carrying out inert gas protection, reacting at 60-80 ℃ in an inert gas atmosphere, stirring for 12-24 hours, after the reaction is finished, adding water to quench the reaction, then adding ethyl acetate, separating an organic phase by using a separating funnel, combining the organic phases, carrying out spin drying, and carrying out column chromatography separation and purification to obtain a target compound (20);
the second step of reaction: sequentially adding the weighed compound (20), hydrazine hydrate and solvent into a reaction bottle, reacting at 60-120 ℃, stirring for 12-36 hours, adding water to quench the reaction after the reaction is finished, then adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, performing column chromatography separation and purification after spin-drying to obtain a target compound (21);
the synthesis method of the compound (23) comprises the following specific steps:
the first step of reaction: sequentially adding the weighed compound (17), a palladium catalyst, alkali and (3- (methoxycarbonyl) phenyl) boric acid into a reaction bottle, then adding a reaction solvent, then carrying out inert gas protection, reacting at 60-80 ℃ in an inert gas atmosphere, stirring for 12-24 hours, after the reaction is finished, adding water to quench the reaction, then adding ethyl acetate, separating an organic phase by using a separating funnel, combining the organic phases, carrying out spin drying, and carrying out column chromatography separation and purification to obtain a target compound (22);
the second step of reaction: and (2) sequentially adding the weighed compound (22), hydrazine hydrate and a solvent into a reaction bottle, reacting at 60-120 ℃, stirring for 12-36 hours, adding water to quench the reaction after the reaction is finished, adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, performing column chromatography separation and purification after spin-drying to obtain the target compound (23).
10. The use of the novel estrogen receptor targeted inhibitor for the treatment of breast cancer according to any one of claims 1 to 9, wherein the test method for the inhibitory activity of the compound (23) on breast cancer cells comprises the following steps:
MCF-7 cell line in logarithmic growth phase is planted in 96-well plate to make each well contain about 5X 103Cells, thermostated at 37 ℃ and containing 5% CO2Culturing in an incubator, pre-dissolving a compound (23) in DMSO to prepare a mother solution with a concentration of 10mg/mL, filtering the mother solution with ultraviolet light, diluting the mother solution with a serum-free culture medium to obtain a compound concentration of 2.5, 5, 10, 20 and 40 mug/mL, adding 100 muL of complete culture medium into each well of a blank control group after cells are completely attached to the wall, adding 100 muL of compounds with different concentrations into each well of an experimental group, arranging 3 multiple wells in parallel in each group, placing the groups in a constant temperature incubator to culture for 48 hours respectively, adding 10 muL of MTT into each well to continue culturing for 4 hours, discarding the culture medium, adding 100 muL of DMSO into each well, placing the 96 well plate on a shaking bed for about 10 minutes in order to fully dissolve purple crystals in the culture medium, finally measuring the absorbance of each well at a wavelength of 570nm by a microplate reader, recording and calculating the survival rate and half-number inhibition concentration of MCF-7, thereby calculating the inhibitory activity of the novel estrogen receptor targeted inhibitor on breast cancer cells.
Technical Field
The invention relates to the technical field of pharmaceutical chemical application, in particular to a synthesis method of a novel estrogen receptor targeting inhibitor and application of the novel estrogen receptor targeting inhibitor in breast cancer treatment.
Background
Breast cancer is the most common malignancy in women worldwide and is also the leading cause of cancer-related deaths in women, and according to the latest survey data of the international agency for research on cancer (IARC), the incidence of breast cancer in women worldwide is 24.2%, with women's cancer first, and 52.9% occurring in developing countries. In china, over 30 million women are diagnosed with breast cancer each year and have an increasing trend year by year. The incidence of breast cancer has risen dramatically in the eastern coastal areas and in economically developed metropolitan areas. Therefore, breast cancer has become the first killer to seriously harm women's health. To date, scientists have not identified the precise cause of breast cancer, but have discovered a number of high-risk factors associated with the onset of breast cancer. Of these, estrone and estradiol are directly associated with the incidence of breast cancer. Thus, endocrine therapy has become a common clinical strategy to prevent cancer cell growth by removing or blocking the action of hormones. Compared with chemotherapy, endocrine therapy has the advantages of definite curative effect, low toxicity, convenient use, no need of hospitalization, easy acceptance by patients and the like. Although the onset is slow, the remission period is long. Is especially suitable for hormone receptor (ER/PR) positive patients.
Among the drugs commonly used in endocrine therapy are antiestrogens and aromatase inhibitors. Wherein the antiestrogen comprises tamoxifen and toremifene. The aromatase inhibitor mainly comprises letrozole, exemestane and the like. However, it was later discovered that antiestrogens and aromatase inhibitors have significant side effects, and in particular tamoxifen is listed as a carcinogen by the world health organization international cancer research institute. Thus, scientists are beginning to search for new therapeutic drugs for breast cancer. Subsequently, lonidamine has been found to have not only contraceptive effect, but also as a treatment for breast cancer. Can inhibit the activity of hexokinase of mitochondria and inhibit oxygen consumption and aerobic glycolysis by changing the ultrastructure of mitochondria, thereby influencing the energy metabolism of cancer cells and achieving the effect of inhibiting the cancer cells. As a thermosensitive drug for treating tumors, it can inhibit cellular respiration under the condition of critical energy requirement. Simultaneously overcomes the side effects of the traditional medicines such as bone marrow suppression and the like.
However, the low bioavailability of lonidamine limits the clinical application of lonidamine. In order to obtain a drug molecule with higher bioavailability, the modification of the structure of lonidamine and the study of structure-activity relationship are tried to obtain a drug molecule with better curative effect for treating breast cancer.
Disclosure of Invention
The invention aims to provide a synthesis method of a novel estrogen receptor targeting inhibitor and application of the novel estrogen receptor targeting inhibitor in breast cancer treatment, and provides novel drug molecules with higher inhibitory activity on breast cancer cells.
In order to achieve the purpose, the invention adopts the following technical scheme: a method for synthesizing a novel estrogen receptor targeting inhibitor,
the synthetic route of the synthetic method of the compound 4 is as follows:
the synthesis method comprises the following steps:
in the first step of reaction, indazole formate 1 and m-tri-substituted benzyl chloride 2 are used as raw materials, alkali in the reaction is one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate and dipotassium hydrogen phosphate, a solvent is one of acetone, cyclohexanone, acetonitrile, tetrahydrofuran and 1, 4-epoxy six-ring, the reaction is carried out in the air, the reaction temperature is 60-80 ℃, and the reaction time is 6-24 hours;
in the second step of reaction, the base used in the reduction reaction of the compound 3 is one of sodium hydroxide and potassium hydroxide, the solvent is one of methanol/water, ethanol/water, tetrahydrofuran/water and acetone/water, the temperature is any temperature between room temperature and 60 ℃, and the reaction time is any time between 12 and 36 hours.
Further, the synthesis method comprises the following specific steps:
the first step of reaction: sequentially adding weighed indazole formate 1, alkali and m-tri-substituted benzyl chloride 2 into a reaction bottle, adding a solvent, reacting at 60-80 ℃, stirring for 6-24 hours, adding water to quench the reaction after the reaction is finished, then adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, and performing column chromatography separation and purification to obtain a target compound 3 after the organic phases are dried in a spinning mode;
the second step of reaction: and sequentially adding the weighed compound 3, alkali and solvent into a reaction bottle, reacting at room temperature to 60 ℃ in an inert gas atmosphere, stirring for 12-36 hours, adding water to quench the reaction after the reaction is finished, adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, drying by spinning, and performing column chromatography separation and purification to obtain the target compound 4.
Further, in the first reaction step,
the alkali is one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate and dipotassium hydrogen phosphate, and the dosage range of the alkali is as follows: 200 and 500mol percent;
the ratio of the dosage of the alkali to the dosage of the compound 1 is as follows: 2:1-5:1, the amount of the base is 1-2 times of the amount of the compound 1;
the position of methyl formate in said compound 1 can be at any of the 3-, 4-, 5-, 6-and 7-positions of indazole;
the position of methyl formate in said compound 3 may be at any of the 3-, 4-, 5-, 6-and 7-positions of indazole;
the position of the formate in the compound 4 may be at any of the 3-, 4-, 5-, 6-and 7-positions of the indazole;
r in the Compound 21Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group; r2Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group; r3Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group;
r in the Compound 31Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group; r2Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group; r3Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group;
r in said Compound 41Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group; r2Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group; r3Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group;
the molar part ratio range of the compound 1 to the compound 2 is as follows: 1:2-1: 1;
the solvent comprises acetone, cyclohexanone, acetonitrile, tetrahydrofuran and 1, 4-epoxy hexacyclic ring;
the reaction temperature is 60-80 ℃;
the reaction time is any time within 6 to 24 hours;
further, in the second-step reaction,
the alkali is one of sodium hydroxide and potassium hydroxide, and the dosage range of the alkali is as follows: 100-200 mol%;
the ratio of the used amount of the alkali to the used amount of the compound 3 is as follows: 1:1-2:1, the amount of the base being 1-2 times the amount of the compound 1;
the solvent comprises methanol/water, ethanol/water, tetrahydrofuran/water and acetone/water;
the reaction temperature is any temperature between room temperature and 60 ℃;
the temperature is any temperature between room temperature and 60 ℃;
the synthetic route of the synthetic method of the compound 5 is as follows:
the synthesis method comprises the following steps:
in the first step of reaction, indazole formate 1 and m-tri-substituted benzyl chloride 2 are used as raw materials, alkali in the reaction is one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate and dipotassium hydrogen phosphate, a solvent is one of acetone, cyclohexanone, acetonitrile, tetrahydrofuran and 1, 4-epoxy six-ring, the reaction is carried out in the air, the reaction temperature is 60-80 ℃, and the reaction time is 6-24 hours;
in the second step of reaction, hydrazine hydrate is used as the hydrazino reagent, one of ethanol, methanol and isopropanol is used as the solvent, the temperature is any temperature between 60 ℃ and 120 ℃, and the reaction time is any time between 2 hours and 6 hours.
Further, the synthesis method comprises the following specific steps:
the first step of reaction: sequentially adding weighed indazole formate 1, alkali and m-tri-substituted benzyl chloride 2 into a reaction bottle, adding a solvent, reacting at 60-80 ℃, stirring for 6-24 hours, adding water to quench the reaction after the reaction is finished, then adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, and performing column chromatography separation and purification to obtain a target compound 3 after the organic phases are dried in a spinning mode;
the second step of reaction: and sequentially adding the weighed compound 3, hydrazine hydrate and a solvent into a reaction bottle, reacting at 60-120 ℃ for 12-36 hours under stirring, adding water to quench the reaction after the reaction is finished, adding ethyl acetate, separating an organic phase by using a separating funnel, combining the organic phases, and performing column chromatography separation and purification after spin-drying to obtain the target compound 5.
Further, in the first reaction step,
the alkali is one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate and dipotassium hydrogen phosphate, and the dosage range of the alkali is as follows: 200 and 500mol percent;
the position of methyl formate in said compound 1 can be at any of the 3-, 4-, 5-, 6-and 7-positions of indazole;
the position of methyl formate in said compound 3 may be at any of the 3-, 4-, 5-, 6-and 7-positions of indazole;
the position of the hydrazide in the compound 5 can be at any of the 3-, 4-, 5-, 6-and 7-positions of the indazole;
r in the Compound 21Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group; r2Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group; r3Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group;
r in the Compound 31Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group; r2Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group; r3Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group;
r in said Compound 51Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group; r2Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group; r3Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group;
the ratio of the dosage of the alkali to the dosage of the compound 1 is as follows: 2:1-5:1, the amount of the base substance is 2-5 times of the amount of the compound (1) substance;
the molar part ratio range of the compound 1 to the compound 2 is as follows: 1:2-1: 1;
the solvent comprises acetone, cyclohexanone, acetonitrile, tetrahydrofuran and 1, 4-epoxy hexacyclic ring;
the reaction temperature is 60-80 ℃;
the reaction time is any time within 6 to 24 hours;
further, in the second-step reaction,
the alkali is hydrazine hydrate, and the dosage range of the hydrazine hydrate is as follows: 100-200 mol%;
the dosage of the hydrazine hydrate and the dosage ratio of the compound 3 are as follows: 1:1-2: 1;
the solvent comprises methanol, ethanol and isopropanol;
the reaction temperature is any temperature between 60 ℃ and 120 ℃;
the reaction time is any time between 2 and 6 hours;
the synthetic route of the synthetic method of the compound 8 is as follows:
the synthesis method comprises the following steps:
in the first step of reaction, 2H-indazole-5-carboxylic acid methyl ester 6 and 2, 4-dichloro-1- (chloromethyl) benzene 2a are used as raw materials, alkali in the reaction is one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate and dipotassium hydrogen phosphate, a solvent is one of acetone, cyclohexanone, acetonitrile, tetrahydrofuran and 1, 4-epoxy hexacyclic ring, the reaction is carried out in the air, the reaction temperature is 60-80 ℃, and the reaction time is 6-24 hours;
in the second step of reaction, hydrazine hydrate is used as the hydrazino reagent, one of ethanol, methanol and isopropanol is used as the solvent, the temperature is any temperature between 60 ℃ and 120 ℃, and the reaction time is any time between 2 hours and 6 hours.
Further, the synthesis method comprises the following specific steps:
the first step of reaction: adding weighed 2H-indazole-5-carboxylic acid methyl ester 6, alkali and 2, 4-dichloro-1- (chloromethyl) benzene 2a into a reaction bottle in sequence, reacting, stirring at 60-80 ℃ for 6-24 hours, adding water to quench the reaction after the reaction is finished, adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, performing column chromatography separation and purification after spin drying to obtain a target compound 7;
the second step of reaction: and sequentially adding the weighed compound 7, hydrazine hydrate and a solvent into a reaction bottle, reacting at 60-120 ℃ for 2-6 hours under stirring, adding water to quench the reaction after the reaction is finished, adding ethyl acetate, filtering the solid by using a suction filter funnel, separating an organic phase by using a separating funnel, combining the organic phases, drying by spinning, and performing column chromatography separation and purification to obtain the target compound 8.
Further, in the first reaction step,
the alkali is one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate and dipotassium hydrogen phosphate, and the dosage range of the alkali is as follows: 200 and 500mol percent;
the ratio of the dosage of the alkali to the dosage of the compound 1 is as follows: 2:1-5:1, the amount of the base is 2-5 times of the amount of the compound 1;
the molar part ratio range of the compound 1 to the compound 2 is as follows: 1:2-1: 1;
the solvent comprises acetone, cyclohexanone, acetonitrile, tetrahydrofuran and 1, 4-epoxy hexacyclic ring;
the reaction temperature is 60-80 ℃;
the reaction time is any time within 6 to 24 hours;
further, in the second-step reaction,
the alkali is hydrazine hydrate, and the dosage range of the hydrazine hydrate is as follows: 100-200 mol%;
the dosage of the hydrazine hydrate and the dosage ratio of the compound 7 are as follows: 1:1-2: 1;
the solvent comprises methanol, ethanol and isopropanol;
the reaction temperature is any temperature between 60 ℃ and 120 ℃;
the reaction time is any time between 2 and 6 hours;
the synthetic route of the synthetic method of the compound 9 is as follows:
the synthesis method comprises the following steps:
in the first step of reaction, 1- (2, 4-dichlorobenzyl) -1H-indazole-3-carboxylic acid 4a is used as a raw material, a chlorination reagent in the reaction is one of thionyl chloride and oxalyl chloride, a solvent is one of tetrahydrofuran, 1, 4-dioxane, acetonitrile and acetone, the reaction is carried out in inert gas, the reaction temperature is between room temperature and 50 ℃, and the reaction time is 2 hours;
in the second step of reaction, the amination reagent is ammonia water, the temperature is room temperature, and the reaction time is any time between 10 and 30 minutes.
Further, the synthesis method comprises the following specific steps:
the first step of reaction: sequentially adding weighed 1- (2, 4-dichlorobenzyl) -1H-indazole-3-carboxylic acid 4a and a solvent into a reaction bottle, then carrying out inert gas protection, slowly dropwise adding a chlorinating reagent at 0 ℃, reacting in an inert gas atmosphere, stirring for 1-3 hours at room temperature to 50 ℃, adding water to quench reaction after the reaction is finished, then adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, and directly carrying out the next reaction after spin-drying;
the second step of reaction: and (3) sequentially adding the crude product and ammonia water in the last step into a reaction bottle, reacting in an inert gas atmosphere, stirring at room temperature for 10-30 minutes, adding water to quench the reaction after the reaction is finished, then adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, performing column chromatography separation and purification after spin drying to obtain the target compound 9.
Further, in the first reaction step,
the chlorination reagent is one of thionyl chloride and oxalyl chloride, and the dosage range of the chlorination reagent is as follows: 2.5-5 mol%;
the dosage of the chlorinating agent and the material ratio of the compound 4a are as follows: 1:1-2:1, the amount of the substance of the chlorinating reagent is 1-2 times of the amount of the substance of the compound 1;
the solvent comprises tetrahydrofuran, 1, 4-dioxane, acetonitrile and acetone;
reacting in the inert gas atmosphere, wherein the inert gas comprises nitrogen, helium and argon;
the reaction temperature is any one of room temperature to 50 ℃;
the reaction time is any time within 6 to 24 hours;
further, in the second-step reaction,
the amination reagent is ammonia water, and the dosage range of the ammonia water is as follows: 100-200 mol%;
the dosage of the ammonia water and the dosage ratio of the compound 4a are as follows: 1:1-2: 1;
the reaction temperature is room temperature;
the reaction time is any time between 10 and 30 minutes;
the synthetic route of the synthetic method of the compound 13 is as follows:
the synthesis method comprises the following steps:
in the first step of reaction, 1H-indole-3-carboxylic acid methyl ester 10 and 2, 4-dichloro-1- (chloromethyl) benzene 2a are used as raw materials, alkali in the reaction is one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate and dipotassium hydrogen phosphate, a solvent is one of acetone, cyclohexanone, acetonitrile, tetrahydrofuran and 1, 4-epoxy hexacyclic ring, the reaction is carried out in the air, the reaction temperature is 60-80 ℃, and the reaction time is 6-24 hours;
in the second step of reaction, hydrazine hydrate is used as the hydrazino reagent, one of ethanol, methanol and isopropanol is used as the solvent, the temperature is any temperature between 60 ℃ and 120 ℃, and the reaction time is any time between 2 hours and 6 hours.
Further, the synthesis method comprises the following specific steps:
the first step of reaction: sequentially adding weighed 1H-indole-3-carboxylic acid methyl ester 10, alkali and 2, 4-dichloro-1- (chloromethyl) benzene 2a into a reaction bottle, adding a reaction solvent, reacting, stirring at 60-80 ℃ for 6-24 hours, adding water to quench and react after the reaction is finished, then adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, drying by spinning, and performing column chromatography separation and purification to obtain a target compound 11;
the second step of reaction: and sequentially adding the weighed compound 11, hydrazine hydrate and a solvent into a reaction bottle, then carrying out inert gas protection, stirring for 2-6 hours at the temperature of 60-120 ℃ for reaction, adding water to quench the reaction after the reaction is finished, then adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, carrying out spin drying, and carrying out column chromatography separation and purification to obtain the target compound 13.
Further, in the first reaction step,
the alkali is one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate and dipotassium hydrogen phosphate, and the dosage range of the alkali is as follows: 200 and 500mol percent;
the ratio of the dosage of the alkali to the dosage of the compound 10 is as follows: 2:1-5:1, the amount of the base is 2-5 times of the amount of the compound 1;
the molar part ratio of the compound 110 to the compound 2a is as follows: 1:2-1: 1;
the solvent comprises acetone, cyclohexanone, acetonitrile, tetrahydrofuran and 1, 4-epoxy hexacyclic ring;
the reaction temperature is 60-80 ℃;
the reaction time is any time within 6 to 24 hours;
further, in the second-step reaction,
the alkali is hydrazine hydrate, and the dosage range of the hydrazine hydrate is as follows: 100-200 mol%;
the dosage of the hydrazine hydrate and the dosage ratio of the compound 11 are as follows: 1:1-2: 1;
the solvent comprises methanol, ethanol and isopropanol;
the reaction temperature is any temperature between 60 ℃ and 120 ℃;
the reaction time is any time between 2 and 6 hours;
the synthetic route of the synthetic method of the compound 14 is as follows:
the synthesis method comprises the following steps:
in the first step of reaction, 1H-indole-3-carboxylic acid methyl ester 10 and 2, 4-dichloro-1- (chloromethyl) benzene 2a are used as raw materials, alkali in the reaction is one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate and dipotassium hydrogen phosphate, a solvent is one of acetone, cyclohexanone, acetonitrile, tetrahydrofuran and 1, 4-epoxy hexacyclic ring, the reaction is carried out in the air, the reaction temperature is 60-80 ℃, and the reaction time is 6-24 hours;
in the second step of reaction, the alkali used in the reduction reaction of the compound 3 is one of sodium hydroxide and potassium hydroxide, the solvent is one of methanol/water, ethanol/water, tetrahydrofuran/water and acetone/water, the temperature is any temperature between room temperature and 60 ℃, and the reaction time is any time between 12 and 36 hours;
in the third step of reaction, the chlorination reagent used in the reaction of the compound 12 is one of thionyl chloride and oxalyl chloride, the solvent is one of tetrahydrofuran, 1, 4-dioxane, acetonitrile and acetone, the reaction is carried out in inert gas, the reaction temperature is between room temperature and 50 ℃, and the reaction time is 2 hours.
In the fourth step of reaction, the amination reagent is ammonia water, the temperature is room temperature, and the reaction time is any time between 10 and 30 minutes.
Further, the synthesis method comprises the following specific steps:
the first step of reaction: sequentially adding weighed 1H-indole-3-carboxylic acid methyl ester 10, alkali and 2, 4-dichloro-1- (chloromethyl) benzene 2a into a reaction bottle, then adding a reaction solvent, reacting, stirring for 6-24 hours at 60-80 ℃, adding water to quench and react after the reaction is finished, then adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, drying by spinning, and performing column chromatography separation and purification to obtain a target compound 11;
the second step of reaction: sequentially adding the weighed compound 11, alkali and solvent into a reaction bottle, reacting in an inert gas atmosphere at room temperature-60 ℃, stirring for 12-36 hours, adding water to quench the reaction after the reaction is finished, then adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, and performing column chromatography separation and purification to obtain a target compound 12 after the organic phases are dried in a spinning mode;
the third step of reaction: and sequentially adding the weighed compound 12 and a solvent into a reaction bottle, then carrying out inert gas protection, slowly dropwise adding a chlorination reagent at 0 ℃, reacting in an inert gas atmosphere at 50 ℃ for 2 hours, adding water to quench the reaction after the reaction is finished, then adding ethyl acetate, separating an organic phase by using a separating funnel, combining the organic phases, and directly carrying out the next reaction after spin-drying.
And a fourth step of reaction: and (3) sequentially adding the crude product and ammonia water in the last step into a reaction bottle, reacting in an inert gas atmosphere, stirring at room temperature for 10-30 minutes, adding water to quench the reaction after the reaction is finished, then adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, performing column chromatography separation and purification after spin drying to obtain the target compound 14.
Further, in the first reaction step,
the alkali is one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate and dipotassium hydrogen phosphate, and the dosage range of the alkali is as follows: 200 and 500mol percent;
the ratio of the dosage of the alkali to the dosage of the compound 10 is as follows: 2:1-5:1, the amount of the base is 2-5 times of the amount of the compound 1;
the molar part ratio of the compound 110 to the compound 2a is as follows: 1:2-1: 1;
the solvent comprises acetone, cyclohexanone, acetonitrile, tetrahydrofuran and 1, 4-epoxy hexacyclic ring;
the reaction temperature is 60-80 ℃;
the reaction time is any time within 6 hours to 24 hours.
Further, in the second-step reaction,
the alkali is one of sodium hydroxide and potassium hydroxide, and the dosage range of the alkali is as follows: 100-200 mol%;
the ratio of the dosage of the alkali to the dosage of the compound 1 is as follows: 1:1-2:1, the amount of the base being 1-2 times the amount of the compound 1;
the solvent comprises methanol/water, ethanol/water, tetrahydrofuran/water and acetone/water;
the reaction temperature is any temperature between room temperature and 60 ℃;
the temperature is any temperature between room temperature and 60 ℃;
the reaction time is any time between 12 and 36 hours;
further, in the third reaction step,
the chlorination reagent is one of thionyl chloride and oxalyl chloride, and the dosage range of the chlorination reagent is as follows: 2.5-5 mol%;
the dosage of the chlorinating agent and the material ratio of the compound 4a are as follows: 1:1-2:1, the amount of the substance of the chlorinating reagent is 1-2 times of the amount of the substance of the compound 1;
the solvent comprises tetrahydrofuran, 1, 4-dioxane, acetonitrile and acetone;
reacting in the inert gas atmosphere, wherein the inert gas comprises nitrogen, helium and argon;
the reaction temperature is any one of room temperature to 50 ℃;
the reaction time is any time within 6 to 24 hours;
further, in the fourth reaction step,
in the fourth reaction step in the synthesis of compound 14,
the amination reagent is ammonia water, and the dosage range of the ammonia water is as follows: 100-200 mol%;
the ratio of the dosage of the ammonia water to the dosage of the compound 12 is as follows: 1:1-2: 1;
the reaction temperature is room temperature;
the reaction time is any time between 10 and 30 minutes.
The synthetic route of the synthetic method of the compound 19 is as follows:
the synthesis method comprises the following steps:
in the first step of reaction, indazole 15 is used as a raw material, alkali in the reaction is one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate and dipotassium hydrogen phosphate, an iodinating reagent is iodine, a reaction solvent is N, N-dimethylformamide, and the reaction is carried out in the air at room temperature;
in the second step of reaction, the alkali used in the coupling reaction between the compound 16 and the 2, 4-dichloro-1- (chloromethyl) benzene 2a is one of cesium carbonate, potassium carbonate, sodium carbonate and silver carbonate, the solvent is one of acetone, cyclohexanone, acetonitrile, tetrahydrofuran and 1, 4-epoxy hexacyclic ring, the temperature is any one of 60 ℃ to 80 ℃, and the reaction time is any one of 6 hours to 24 hours;
in the third step of reaction, the catalyst used in the coupling reaction of the compound 17 and allyl formate is one of palladium acetate, palladium chloride, palladium bromide and palladium trifluoroacetate, the base is one of N, N-diisopropylethylamine and triethylamine, the phosphine ligand is one of triphenylphosphine, tricyclohexylphosphine and 1, 2-bis (diphenylphosphino) ethane, the reaction solvent is N, N-dimethylformamide, the temperature in the reaction is any one of 100-120 ℃, and the reaction time is any one time between 12-24 hours.
In the fourth step of reaction, hydrazine hydrate is used as a hydrazinolysis reagent, one of ethanol, methanol and isopropanol is used as a solvent, the temperature is any one of 60-120 ℃, and the reaction time is any one of 2-6 hours.
Further, the synthesis method comprises the following specific steps:
the first step of reaction: adding the weighed indazole 15, iodination reagent and solvent into a reaction bottle in sequence, reacting, stirring at room temperature for 1-3 hours, adding water to quench the reaction after the reaction is finished, adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, performing column chromatography separation and purification after spin drying to obtain a target compound 16;
the second step of reaction: sequentially adding the weighed compound 16, alkali and 2, 4-dichloro-1- (chloromethyl) benzene 2a into a reaction bottle, then adding a reaction solvent, reacting at 60-80 ℃ for stirring for 6-24 hours, adding water to quench the reaction after the reaction is finished, then adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, carrying out spin drying, and carrying out column chromatography separation and purification to obtain a target compound 17;
the third step of reaction: adding the weighed compound 17, a palladium catalyst, alkali, a phosphine ligand and allyl formate into a reaction bottle in sequence, then adding a reaction solvent, then carrying out inert gas protection, stirring for 12-24 hours at the temperature of 100-120 ℃ for reaction, adding water to quench the reaction after the reaction is finished, then adding ethyl acetate, filtering the solid by using a suction filter funnel, then separating the organic phase by using a separating funnel, combining the organic phases, carrying out spin drying, and carrying out column chromatography separation and purification to obtain the target compound 18.
And a fourth step of reaction: and sequentially adding the weighed compound 18, hydrazine hydrate and a solvent into a reaction bottle, then carrying out inert gas protection, stirring for 2-6 hours at the temperature of 60-120 ℃ for reaction, adding water to quench the reaction after the reaction is finished, then adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, carrying out spin drying, and carrying out column chromatography separation and purification to obtain the target compound 19.
Further, in the first reaction step,
the alkali is one of sodium hydroxide and potassium hydroxide, and the dosage range of the alkali is as follows: 100-200 mol%;
the ratio of the dosage of the alkali to the dosage of the compound 15 is as follows: 1:1-2:1, the amount of the base being 1-2 times the amount of the compound 1;
the compound 15 with iodine I2The molar ratio ranges are as follows: 1:2-1: 1;
the solvent is N, N-dimethylformamide;
the reaction temperature is room temperature;
the reaction time is any time within 2 to 4 hours;
further, in the second-step reaction,
the alkali is one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate and dipotassium hydrogen phosphate, and the dosage range of the alkali is as follows: 200 and 500mol percent;
the ratio of the dosage of the alkali to the dosage of the compound 10 is as follows: 2:1-5:1, the amount of the base is 2-5 times of the amount of the compound 1;
the molar part ratio of the compound 110 to the compound 2a is as follows: 1:2-1: 1;
the solvent comprises acetone, cyclohexanone, acetonitrile, tetrahydrofuran and 1, 4-epoxy hexacyclic ring;
the reaction temperature is 60-80 ℃;
the reaction time is any time within 6 to 24 hours;
further, in the third reaction step,
the metal palladium catalyst is one of palladium acetate, palladium chloride, palladium bromide and palladium trifluoroacetate, and the dosage range of the catalyst is as follows: 10-20 mol%;
the phosphine ligand is one of triphenylphosphine, tricyclohexylphosphine and 1, 2-bis (diphenylphosphino) ethane, and the dosage range of the phosphine ligand is as follows: 25-50 mol%;
the alkali is one of N, N-diisopropylethylamine and triethylamine, and the dosage range of the alkali is as follows: 200-300 mol%;
the molar part ratio of the compound 17 to the allyl formate is as follows: 1:2-1: 5;
the reaction solvent is N, N-dimethylformamide;
reacting in the inert gas atmosphere, wherein the inert gas comprises nitrogen, helium and argon;
the temperature in the reaction is any one of 100 ℃ to 120 ℃,
the reaction time is any time between 12 and 24 hours.
As a preparation method of the novel estrogen receptor targeting inhibitor and the application thereof in the treatment of breast cancer provided by the invention, in the fourth step reaction,
the alkali is hydrazine hydrate, and the dosage range of the hydrazine hydrate is as follows: 100-200 mol%;
the dosage of the hydrazine hydrate and the dosage ratio of the compound 18 are as follows: 1:1-2: 1;
the solvent comprises methanol, ethanol and isopropanol;
the reaction temperature is any temperature between 60 ℃ and 120 ℃;
the reaction time is any time between 2 and 6 hours.
The synthetic route of the synthetic method of the compound (21) is as follows:
the synthesis method comprises the following steps:
in the first step of reaction, compound 17 and (4- (methoxycarbonyl) phenyl) boric acid are used as raw materials, a metal catalyst in the reaction is one of palladium acetate, palladium chloride, palladium bromide and palladium trifluoroacetate, alkali is one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate and dipotassium hydrogen phosphate, a reaction solvent is a mixed solvent of ethanol/toluene/water, and the reaction is carried out in an inert gas at any temperature of 80-120 ℃;
in the second step of reaction, hydrazine hydrate is used as the hydrazino reagent, one of ethanol, methanol and isopropanol is used as the solvent, the temperature is any temperature between 60 ℃ and 120 ℃, and the reaction time is any time between 2 hours and 6 hours.
Further, the synthesis method comprises the following specific steps:
the first step of reaction: sequentially adding the weighed compound 17, a palladium catalyst, alkali and (4- (methoxycarbonyl) phenyl) boric acid into a reaction bottle, then adding a reaction solvent, carrying out inert gas protection, reacting at 80-120 ℃ in an inert gas atmosphere, stirring for 12-24 hours, after the reaction is finished, adding water to quench the reaction, then adding ethyl acetate, separating an organic phase by using a separating funnel, combining the organic phases, carrying out spin drying, and carrying out column chromatography separation and purification to obtain a target compound 20;
the second step of reaction: and sequentially adding the weighed compound 20, hydrazine hydrate and solvent into a reaction bottle, reacting at 60-120 ℃ for 2-6 hours under stirring, adding water to quench the reaction after the reaction is finished, adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, performing column chromatography separation and purification after spin drying to obtain the target compound 21.
Further, in the first reaction step,
the metal palladium catalyst is one of palladium acetate, palladium chloride, palladium bromide and palladium trifluoroacetate, and the dosage range of the catalyst is as follows: 10-20 mol%;
the alkali is one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate and dipotassium hydrogen phosphate, and the dosage range of the alkali is as follows: 100-200 mol%;
the ratio of the used amount of the alkali to the used amount of the compound 17 is as follows: 1:1-2:1, the amount of the base being 1-2 times the amount of the compound 1;
the molar ratio of the compound 17 to the (4- (methoxycarbonyl) phenyl) boric acid is as follows: 1:2-1: 1;
the solvent is a mixed solvent of ethanol/toluene/water;
reacting in the inert gas atmosphere, wherein the inert gas comprises nitrogen, helium and argon;
the reaction temperature is any one of 80-120 ℃;
the reaction time is any time within 12 hours to 24 hours;
further, in the second-step reaction,
the alkali is hydrazine hydrate, and the dosage range of the hydrazine hydrate is as follows: 100-200 mol%;
the dosage of the hydrazine hydrate and the dosage ratio of the compound 20 are as follows: 1:1-2: 1;
the solvent comprises methanol, ethanol and isopropanol;
the reaction temperature is any temperature between 60 ℃ and 120 ℃;
the reaction time is any time between 2 and 6 hours.
The synthetic route of the synthetic method of the compound 23 is as follows:
the synthesis method comprises the following steps:
in the first step of reaction, compound 17 and (3- (methoxycarbonyl) phenyl) boric acid are used as raw materials, a metal catalyst in the reaction is one of palladium acetate, palladium chloride, palladium bromide and palladium trifluoroacetate, alkali is one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate and dipotassium hydrogen phosphate, a reaction solvent is a mixed solvent of ethanol/toluene/water, and the reaction is carried out in an inert gas at any temperature of 80-120 ℃;
in the second step of reaction, hydrazine hydrate is used as the hydrazino reagent, one of ethanol, methanol and isopropanol is used as the solvent, the temperature is any temperature between 80 ℃ and 150 ℃, and the reaction time is any time between 12 hours and 36 hours.
Further, the synthesis method comprises the following specific steps:
the first step of reaction: sequentially adding the weighed compound 17, a palladium catalyst, alkali and (3- (methoxycarbonyl) phenyl) boric acid into a reaction bottle, then adding a reaction solvent, then carrying out inert gas protection, stirring at room temperature for 1-3 hours in an inert gas atmosphere, after the reaction is finished, adding water to quench the reaction, then adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, carrying out spin drying, and carrying out column chromatography separation and purification to obtain a target compound 20;
the second step of reaction: and sequentially adding the weighed compound 22, hydrazine hydrate and solvent into a reaction bottle, reacting at the temperature of 80-150 ℃, stirring for 12-36 hours, adding water to quench the reaction after the reaction is finished, then adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, performing column chromatography separation and purification after spin drying to obtain the target compound 23.
Further, in the first reaction step,
the metal palladium catalyst is one of palladium acetate, palladium chloride, palladium bromide and palladium trifluoroacetate, and the dosage range of the catalyst is as follows: 10-20 mol%;
the alkali is one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate and dipotassium hydrogen phosphate, and the dosage range of the alkali is as follows: 100-200 mol%;
the ratio of the used amount of the alkali to the used amount of the compound 17 is as follows: 1:1-2:1, the amount of the base being 1-2 times the amount of the compound 1;
the molar ratio of the compound 17 to the (3- (methoxycarbonyl) phenyl) boric acid is as follows: 1:2-1: 1;
the solvent is a mixed solvent of ethanol/toluene/water;
and reacting in an inert gas atmosphere, wherein the inert gas comprises nitrogen, helium and argon.
The reaction temperature is any one of 80-120 ℃;
the reaction time is any time within 12 hours to 24 hours;
further, in the second-step reaction,
the alkali is hydrazine hydrate, and the dosage range of the hydrazine hydrate is as follows: 100-200 mol%;
the ratio of the usage amount of the hydrazine hydrate to the usage amount of the compound 22 is as follows: 1:1-2:1,
the solvent comprises methanol, ethanol and isopropanol;
the reaction temperature is any temperature between 60 ℃ and 120 ℃;
the reaction time is any time between 2 and 6 hours.
In order to better achieve the above object, the present invention further provides a use of a novel estrogen receptor targeted inhibitor in the treatment of breast cancer, wherein the method for testing the inhibitory activity of the compound 23 on breast cancer cells comprises the following steps:
taking MCF-7 of logarithmic growth phase and planting in 96-well plate to make each well contain about 5 × 103Cells, thermostated at 37 ℃ and containing 5% CO2Culturing in an incubator. The compounds were pre-dissolved in DMSO, prepared as a stock solution at a concentration of 10mg/mL (DMSO does not affect cell viability), filtered after UV irradiation, and diluted in serum-free medium at compound concentrations of 2.5, 5, 10, 20, 40. mu.g/mL. After the cells are completely attached to the wall, 100 mul of complete culture medium is added into each hole of the blank control group, and each hole of the experimental group is added100 μ L of compounds of different concentrations, each group having 3 multiple wells in parallel, were placed in a thermostated incubator (37 ℃, 5% CO)2) Respectively culturing for 48 h. Subsequently, 10. mu.L of MTT (5mg/mL) was added to each well and the culture was continued for 4h, the medium was discarded, and after 100. mu.L of DMSO was added to each well, the 96-well plate was placed on a shaker for about 10min in order to allow the purple crystals in the medium to be sufficiently dissolved. Finally, the absorbance of each well at a wavelength of 570nm was measured by a microplate reader, and the survival rate and median Inhibitory Concentration (IC) of MCF-7 were recorded and calculated50)。
Compared with the prior art, the invention has the beneficial effects that:
(1) the invention overcomes the defects of large side effect, low bioavailability and the like of the existing medicines, synthesizes and obtains a class of ER-targeted medicine molecules by adopting medicine design based on structure and a computer-aided strategy, shows higher cancer cell inhibition activity than lonidamine, and is expected to become a new medicine molecule for breast cancer replacing lonidamine.
(2) The catalyst used for synthesizing the ER-targeted drug molecules is low in dosage, the effects of simplifying the process, reducing the cost and facilitating the post-treatment process are achieved while the good catalytic effect and the cost are maintained, the recycling rate of the solvent is high, the pollution to the environment is reduced to the minimum, and the effects of green production and zero pollutant emission can be basically realized.
(3) The method for synthesizing the ER-targeted drug molecules has the advantages of short reaction steps, low requirement on equipment, simple operation, capability of improving the reaction yield to a greater extent and further saving the production cost.
(4) The implementation of the medicine of the invention not only can promote the development of micromolecular medicines for more efficiently treating breast cancer diseases, but also can bring great promotion effect to national economy of China.
(6) The invention relates to a preparation method of a novel estrogen receptor targeting inhibitor and application research thereof in breast cancer treatment.
(7) The novel ER targeting inhibitor synthesized by the invention exerts a very strong inhibition effect on the aspect of inhibiting the proliferation of breast cancer cells, has a better inhibition effect on the proliferation of the breast cancer cells than the traditional medicament lonidamine, has the potential of being developed into a novel breast cancer treatment medicament, has simple synthesis steps, does not need to adopt dangerous, flammable and explosive reagents, is convenient to operate, has good product selectivity and high yield, and is suitable for industrial production.
Description of the drawings
FIG. 1 is a graph showing the effect of molecular docking experiments on 4- (1- (2, 4-dichlorobenzyl) -1H-indazol-3-yl) benzoyl hydrazine 21, 3- (1- (2, 4-dichlorobenzyl) -1H-indazol-3-yl) benzoyl hydrazine 23 and estrogen receptor ER in the present invention.
FIG. 2 is a nuclear magnetic hydrogen spectrum of 4- (1- (2, 4-dichlorobenzyl) -1H-indazol-3-yl) benzoyl hydrazine 21 and 3- (1- (2, 4-dichlorobenzyl) -1H-indazol-3-yl) benzoyl hydrazine 23 according to the present invention.
Detailed Description
The following examples further illustrate the present invention but are not to be construed as limiting the invention. Modifications or substitutions to methods, procedures, or conditions of the invention may be made without departing from the spirit and scope of the invention. The experimental methods and reagents of the formulations not specified in the examples are in accordance with the conventional conditions in the art.
Example 1
Synthesis of 1- (2-chloro-4-fluorobenzyl) -1H-indazole-3-carboxylic acid 4 a:
and sequentially adding weighed 0.20mmol of compound 1a, 0.22mmol of 2a and 0.9mmol of potassium carbonate into a reaction bottle, then adding 2mL of acetone, carrying out reflux stirring at 70 ℃ for 12 hours, after the reaction is finished, adding water to quench the reaction, adding 5mL of ethyl acetate to dilute the reaction, washing the reaction with 5mL of saturated saline solution, drying an organic phase by anhydrous magnesium sulfate, carrying out spin drying, and then recrystallizing by using a dichloromethane/n-hexane system to obtain an intermediate 3 a. Subsequently, the intermediate 3a obtained by recrystallization,adding 0.40mmol sodium hydroxide and 2mL methanol/water (1:1) mixed solvent into a reaction bottle, refluxing and stirring at 60 ℃ for 24 hours, adding water to quench the reaction after the reaction is finished, adding 5mL ethyl acetate to dilute, washing with 5mL saturated saline, drying the organic phase with anhydrous magnesium sulfate, performing spin drying, and recrystallizing with a dichloromethane/n-hexane system to obtain a product 4a (IC)50487 μ M) was 54mg of white solid.
1H NMR(400MHz,DMSO)δ13.16(s,1H),8.12(d,J＝8.4Hz,1H),7.81(d,J＝8.4Hz,1H),7.56-7.45(m,2H),7.34(t,J＝7.6Hz,1H),7.19(td,J＝8.6,2.4Hz,1H),7.07(dd,J＝8.8,6.4Hz,1H),5.82(s,2H).13C NMR(100MHz,DMSO)δ163.4,162.8,160.3,140.9,135.9,133.2(d,J＝10.6Hz),131.4(d,J＝9.2Hz),130.5(d,J＝3.4Hz),127.0,123.1,121.7,116.9(d,J＝25.1Hz),114.8(d,J＝21.1Hz),110.6,49.7。
Example 2
Synthesis of 1- (2, 4-difluorobenzyl) -1H-indazole-3-carboxylic acid 4 b:
and sequentially adding weighed 0.20mmol of compound 1a, 0.22mmol of compound 2b and 0.9mmol of potassium carbonate into a reaction bottle, then adding 2mL of acetone, carrying out reflux stirring at 70 ℃ for 12 hours, after the reaction is finished, adding water to quench the reaction, adding 5mL of ethyl acetate to dilute the reaction, washing the reaction with 5mL of saturated saline solution, drying an organic phase by anhydrous magnesium sulfate, carrying out spin drying, and then recrystallizing by using a dichloromethane/n-hexane system to obtain an intermediate 3 b. Then adding the intermediate 3b obtained by recrystallization, 0.40mmol of potassium hydroxide and 2mL of methanol/water (1:1) mixed solvent into a reaction bottle, refluxing and stirring at 60 ℃ for 24 hours, adding water to quench the reaction after the reaction is finished, adding 5mL of ethyl acetate to dilute the reaction solution, washing with 5mL of saturated saline solution, drying an organic phase by anhydrous magnesium sulfate, performing spin drying, and then recrystallizing by a dichloromethane/n-hexane system to obtain a product 4b (IC)50199 μ M) as a white solid 51 mg.
1H NMR(400MHz,DMSO)δ8.25(d,J＝8.0Hz,1H),7.55(d,J＝8.4Hz,1H),7.33(t,J＝7.6Hz,1H),7.23(td,J＝10.4,2.4Hz,1H),7.19-7.09(m,2H),6.97(td,J＝8.4,2.0Hz,1H),5.66(s,2H).13C NMR(100MHz,DMSO)δ166.9,162.2(dd,J＝197.8,12.2Hz),159.7(dd,J＝199.2,12.1Hz),143.7,140.4,131.5(dd,J＝9.5,5.3Hz),126.4,123.6,123.3,121.3,120.6(dd,J＝15.0,3.2Hz),111.7(dd,J＝21.2,2.9Hz),109.6,104.0(t,J＝25.7Hz),45.3。
Example 3
Synthesis of 1- (2, 6-dichlorobenzyl) -1H-indazole-3-carboxylic acid 4 c:
and sequentially adding weighed 0.20mmol of compound 1a, 0.22mmol of compound 2c and 0.9mmol of sodium carbonate into a reaction bottle, then adding 2mL of acetone, carrying out reflux stirring at 70 ℃ for 12 hours, after the reaction is finished, adding water to quench the reaction, adding 5mL of ethyl acetate to dilute the reaction, washing the reaction with 5mL of saturated saline solution, drying an organic phase by anhydrous magnesium sulfate, carrying out spin drying, and then recrystallizing by using a dichloromethane/n-hexane system to obtain an intermediate 3 c. Then adding the intermediate 3c obtained by recrystallization, 0.40mmol of sodium hydroxide and 2mL of ethanol/water (1:1) mixed solvent into a reaction bottle, refluxing and stirring at 60 ℃ for 24 hours, adding water to quench the reaction after the reaction is finished, adding 5mL of ethyl acetate to dilute the reaction, washing with 5mL of saturated saline solution, drying an organic phase by anhydrous magnesium sulfate, performing spin drying, and then recrystallizing by a dichloromethane/n-hexane system to obtain a product 4c (IC)50233 μ M) as a white solid 53 mg.
1H NMR(400MHz,DMSO)δ8.08(d,J＝8.0Hz,1H),7.89(d,J＝8.8Hz,1H),7.57(d,J＝1.2Hz,1H),7.55(s,1H),7.52(ddd,J＝8.0,6.8,0.8Hz,1H),7.46(dd,J＝8.8,7.2Hz,1H),7.34(t,J＝7.6Hz,1H),5.90(s,2H).13C NMR(100MHz,DMSO)δ163.4,141.1,136.2,135.6,131.3,131.1,129.0,127.0,123.1,122.9,121.7,110.8,48.1。
Example 4
Synthesis of 1- (2,4, 6-trimethylbenzyl) -1H-indazole-3-carboxylic acid 4 d:
and sequentially adding weighed 0.20mmol of compound 1a, 0.22mmol of compound 2d and 0.9mmol of potassium carbonate into a reaction bottle, then adding 2mL of tetrahydrofuran, carrying out reaction at 70 ℃ under reflux and stirring for 12 hours, after the reaction is finished, adding water to quench the reaction, adding 5mL of ethyl acetate to dilute the reaction, washing the reaction with 5mL of saturated saline, drying an organic phase by anhydrous magnesium sulfate, and recrystallizing the organic phase by a dichloromethane/n-hexane system to obtain an intermediate 3 d. Then adding the intermediate 3d obtained by recrystallization, 0.40mmol of sodium hydroxide and 2mL of methanol/water (1:1) mixed solvent into a reaction bottle, refluxing and stirring at 60 ℃ for 24 hours, adding water to quench the reaction after the reaction is finished, adding 5mL of ethyl acetate to dilute the reaction, washing with 5mL of saturated saline solution, drying an organic phase by anhydrous magnesium sulfate, performing spin drying, and then recrystallizing by a dichloromethane/n-hexane system to obtain a product 4d (IC)50111 μ M) was 50mg of white solid.
1H NMR(400MHz,DMSO)δ8.22(d,J＝8.0Hz,1H),7.35(d,J＝8.8Hz,1H),7.23(t,J＝7.6Hz,1H),7.05(t,J＝7.6Hz,1H),6.86(s,2H),5.51(s,2H),2.21(s,3H),2.19(s,6H).13C NMR(100MHz,DMSO)δ167.0,142.8,140.4,137.7,137.1,129.7,129.2,125.8,123.7,123.4,120.8,109.6,47.8,20.8,20.0。
The invention provides a preparation method of a novel estrogen receptor targeting inhibitor, wherein the reaction route of a compound 5 in the following examples is as follows:
the first step of reaction: sequentially adding weighed indazole formate (1), alkali and m-tri-substituted benzyl chloride (2) into a reaction bottle, adding a solvent, reacting at 60-80 ℃, stirring for 6-24 hours, adding water to quench the reaction after the reaction is finished, then adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, and performing column chromatography separation and purification to obtain a target compound 3 after spin drying;
specifically, the alkali is one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate and dipotassium hydrogen phosphate, and the dosage range of the alkali is as follows: 200 and 500mol percent;
specifically, the position of methyl formate in the compound (1) may be at any one of positions 3, 4, 5, 6 and 7 of indazole;
specifically, the position of methyl formate in the compound (3) may be at any one of positions 3, 4, 5, 6 and 7 of indazole;
specifically, the position of the hydrazide in the compound (5) may be at any one of the 3-, 4-, 5-, 6-and 7-positions of the indazole;
specifically, R in the compound (2)1Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group; r2Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group; r3Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group;
specifically, R in the compound (3)1Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group; r2Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group; r3Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group;
specifically, R in the compound (5)1Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group; r2Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group; r3Is selected from C1-C10Alkyl radical, C6-C12Aryl of (A), C6-C12Heterocyclic aromatic radical, C3-C12Cycloalkyl radical, C3-C12A heterocycloalkyl group;
specifically, the ratio of the used amount of the base to the used amount of the compound (1) is as follows: 2:1-5:1, the amount of the base substance is 2-5 times of the amount of the compound (1) substance;
specifically, the molar part ratio of the compound (1) to the compound (2) is as follows: 1:2-1: 1;
specifically, the solvent comprises acetone, cyclohexanone, acetonitrile, tetrahydrofuran, and 1, 4-epoxy hexacyclic ring;
specifically, the reaction temperature is 60-80 ℃;
specifically, the reaction time is any time within 6 hours to 24 hours;
the second step of reaction: and sequentially adding the weighed compound 3, hydrazine hydrate and a solvent into a reaction bottle, reacting at 60-120 ℃ for 12-36 hours under stirring, adding water to quench the reaction after the reaction is finished, adding ethyl acetate, separating an organic phase by using a separating funnel, combining the organic phases, and performing column chromatography separation and purification after spin-drying to obtain the target compound 5.
Specifically, the alkali is hydrazine hydrate, and the dosage range of the hydrazine hydrate is as follows: 100-200 mol%;
specifically, the ratio of the usage amount of the hydrazine hydrate to the usage amount of the compound (3) is as follows: 1:1-2: 1;
specifically, the solvent includes methanol, ethanol, isopropanol;
specifically, the reaction temperature is any temperature between 60 ℃ and 120 ℃;
in particular, the reaction time is any time between 2 and 6 hours.
Example 5
Synthesis of (1- (2-chloro-4-fluorobenzyl) -1H-indazole-3-carbohydrazide 5 a:
and sequentially adding weighed 0.20mmol of compound 1a, 0.22mmol of compound 2a and 0.9mmol of potassium carbonate into a reaction bottle, then adding 2mL of acetone, carrying out reflux stirring at 70 ℃ for 12 hours, after the reaction is finished, adding water to quench the reaction, adding 5mL of ethyl acetate to dilute the reaction, washing the reaction with 5mL of saturated saline solution, drying an organic phase by anhydrous magnesium sulfate, carrying out spin drying, and then recrystallizing by using a dichloromethane/n-hexane system to obtain an intermediate 3 a. Then adding the intermediate 3a obtained by recrystallization, 10mmol hydrazine hydrate and 2mL ethanol into a reaction bottle, refluxing and stirring the reaction at 80 ℃ for 4 hours, adding water to quench the reaction after the reaction is finished, adding 5mL ethyl acetate to dilute the reaction, washing the reaction with 5mL saturated saline, drying an organic phase by anhydrous magnesium sulfate, performing spin drying, and recrystallizing by using a dichloromethane/n-hexane system to obtain a product 5a (IC) which is a product50202 μ M) was 56mg of white solid.
1H NMR(400MHz,CDCl3)δ8.38(d,J＝8.0Hz,1H),8.08(s,1H),7.47-7.37(m,2H),7.36-7.29(m,1H),7.18(dd,J＝8.4,2.4Hz,1H),6.87(td,J＝8.4,2.4Hz,1H),6.79(dd,J＝8.5,6.0Hz,1H),5.66(s,2H),4.08(d,J＝4.4Hz,2H).13C NMR(100MHz,CDCl3)δ163.4,162.2(d,J＝249.3Hz),141.0,137.0,133.5(J＝10.3Hz),130.1(J＝8.9Hz),129.7(J＝3.6Hz),127.6,123.3,123.2,122.8,117.2(J＝24.8Hz),114.7(J＝21.2Hz),109.5,50.1。
Example 6
Synthesis of 1- (2, 4-difluorobenzyl) -1H-indazole-3-carbohydrazide 5 b:
and sequentially adding weighed 0.20mmol of compound 1a, 0.22mmol of compound 2b and 0.9mmol of potassium carbonate into a reaction bottle, then adding 2mL of acetone, carrying out reflux stirring at 70 ℃ for 12 hours, after the reaction is finished, adding water to quench the reaction, adding 5mL of ethyl acetate to dilute the reaction, washing the reaction with 5mL of saturated saline solution, drying an organic phase by anhydrous magnesium sulfate, carrying out spin drying, and then recrystallizing by using a dichloromethane/n-hexane system to obtain an intermediate 3 b. Then adding the intermediate 3b obtained by recrystallization, 10mmol hydrazine hydrate and 2mL methanol into a reaction bottle, refluxing and stirring the reaction at 80 ℃ for 4 hours, adding water to quench the reaction after the reaction is finished, adding 5mL ethyl acetate to dilute the reaction, washing the reaction with 5mL saturated saline, drying the organic phase by anhydrous magnesium sulfate, performing spin drying, and recrystallizing by using a dichloromethane/n-hexane system to obtain a product 5b (IC) which is a product 5b (50222 μ M) was 51mg of a white solid.
1H NMR(400MHz,CDCl3)δ8.35(d,J＝8.0Hz,1H),8.10(s,1H),7.49-7.40(m,2H),7.31(ddd,J＝8.4,6.0,1.6Hz,1H),7.12-7.03(m,1H),6.91-6.73(m,2H),5.59(s,2H),4.08(d,J＝4.0Hz,2H).13C NMR(100MHz,CDCl3)δ164.2(d,J＝11.9Hz),163.4,161.7(dd,J＝12.0,2.4Hz),159.2(d,J＝11.9Hz),140.7,136.8,130.8(dd,J＝9.7,5.2Hz),127.5,123.2,123.0(d,J＝42.4Hz),119.0(dd,J＝14.9,3.8Hz),112.0(dd,J＝21.3,3.8Hz),109.4(d,J＝1.5Hz),104.2(t,J＝25.3Hz),46.2(d,J＝4.1Hz)。
Example 7
Synthesis of 1- (2, 6-dichlorobenzyl) -1H-indazole-3-carbohydrazide 5 c:
adding weighed 0.20mmol of compound 1a, 0.22mmol of compound 2c and 0.9mmol of sodium carbonate into a reaction bottle in sequence, then adding 2mL of tetrahydrofuran, reacting at 70 ℃, refluxing and stirring for 12 hours, adding water to quench the reaction after the reaction is finished, adding 5mL of ethyl acetate for dilution, and adding 5mL of saturated saline waterWashing, drying the organic phase with anhydrous magnesium sulfate, spin-drying, and recrystallizing with dichloromethane/n-hexane system to obtain intermediate 3 c. Then adding the intermediate 3c obtained by recrystallization, 10mmol hydrazine hydrate and 2mL ethanol into a reaction bottle, refluxing and stirring the reaction at 80 ℃ for 4 hours, adding water to quench the reaction after the reaction is finished, adding 5mL ethyl acetate to dilute the reaction, washing the reaction with 5mL saturated saline, drying the organic phase by anhydrous magnesium sulfate, performing spin drying, and recrystallizing by using a dichloromethane/n-hexane system to obtain a product 5c (IC)50181 μ M) as a white solid 56 mg.
1H NMR(400MHz,CDCl3)δ8.34(d,J＝8.4Hz,1H),7.99(s,1H),7.55(d,J＝8.8Hz,1H),7.48-7.41(m,1H),7.39(d,J＝7.6Hz,2H),7.33-7.27(m,2H),5.79(s,2H),4.02(d,J＝4.4Hz,2H).13C NMR(100MHz,CDCl3)δ163.4,141.2,137.1,136.5,131.1,130.5,128.8,127.2,122.94,122.85,122.6,109.7,48.3。
Example 8
Synthesis of 1- (2,4, 6-trimethylbenzyl) -1H-indazole-3-carbohydrazide 5 d:
and sequentially adding weighed 0.20mmol of compound 1a, 0.22mmol of compound 2d and 0.9mmol of potassium carbonate into a reaction bottle, then adding 2mL of acetone, carrying out reflux stirring at 70 ℃ for 12 hours, after the reaction is finished, adding water to quench the reaction, adding 5mL of ethyl acetate to dilute the reaction, washing the reaction with 5mL of saturated saline solution, drying an organic phase by anhydrous magnesium sulfate, carrying out spin drying, and then recrystallizing by using a dichloromethane/n-hexane system to obtain an intermediate 3 d. Then adding the intermediate 3d obtained by recrystallization, 10mmol hydrazine hydrate and 2mL isopropanol into a reaction bottle, carrying out reflux stirring at 80 ℃ for 4 hours, adding water to quench the reaction after the reaction is finished, adding 5mL ethyl acetate to dilute the reaction, washing with 5mL saturated saline, drying the organic phase with anhydrous magnesium sulfate, carrying out spin drying, and then recrystallizing with a dichloromethane/n-hexane system to obtain a product 5d (IC) which is a product 5d (50188 μ M) as a white solid 49 mg.
1H NMR(400MHz,CDCl3)δ8.33(d,J＝8.0Hz,1H),8.00(s,1H),7.40-7.33(m,1H),7.32-7.23(m,2H),6.91(s,2H),5.52(s,2H),4.02(d,J＝4.0Hz,2H),2.3(s,3H),2.28(s,6H).13C NMR(100MHz,CDCl3)δ163.6,140.9,138.4,138.0,136.0,129.5,128.6,127.0,123.1,122.8,122.6,109.7,48.1,21.1,20.3。
Example 9
Synthesis of 1- ((perfluorophenyl) methyl) -1H-indazole-3-carbohydrazide 5 e:
and sequentially adding weighed 0.20mmol of compound 1a, 0.22mmol of compound 2e and 0.9mmol of potassium bicarbonate into a reaction bottle, then adding 2mL of acetone, carrying out reflux stirring at 70 ℃ for 12 hours, after the reaction is finished, adding water to quench the reaction, adding 5mL of ethyl acetate to dilute the reaction, washing the reaction with 5mL of saturated saline solution, drying an organic phase by anhydrous magnesium sulfate, carrying out spin drying, and then recrystallizing by using a dichloromethane/n-hexane system to obtain an intermediate 3 e. Then adding the intermediate 3e obtained by recrystallization, 10mmol hydrazine hydrate and 2mL ethanol into a reaction bottle, refluxing and stirring the reaction at 80 ℃ for 4 hours, adding water to quench the reaction after the reaction is finished, adding 5mL ethyl acetate to dilute the reaction, washing the reaction with 5mL saturated saline, drying the organic phase by anhydrous magnesium sulfate, performing spin drying, and recrystallizing by using a dichloromethane/n-hexane system to obtain a product 5e (IC)50115 μ M) as a white solid 52 mg.
1H NMR(400MHz,CDCl3)δ8.35(d,J＝8.4Hz,1H),8.06(s,1H),7.56(d,J＝8.4Hz,1H),7.50(t,J＝7.6Hz,1H),7.33(t,J＝7.4Hz,1H),5.63(s,2H),4.07(d,J＝4.0Hz,2H).13C NMR(100MHz,CDCl3)δ163.1,145.7(dm,J＝248.6Hz),141.8(dm,J＝254.9Hz),140.7,137.7(dm,J＝249.6Hz),137.4,127.8,123.3,123.0,122.9,109.3(td,J＝17.6,4.0Hz),109.1,40.3。
Example 10
Synthesis of 1- (2- (trifluoromethyl) benzyl) -1H-indazole-3-carbohydrazide 5 f:
and sequentially adding weighed 0.20mmol of compound 1a, 0.22mmol of compound 2f and 0.9mmol of potassium carbonate into a reaction bottle, then adding 2mL of acetonitrile, carrying out reflux stirring at 80 ℃ for 18 hours, after the reaction is finished, adding water to quench the reaction, adding 5mL of ethyl acetate to dilute the reaction, washing the reaction with 5mL of saturated saline solution, drying an organic phase by anhydrous magnesium sulfate, carrying out spin drying, and then recrystallizing by using a dichloromethane/n-hexane system to obtain an intermediate 3 f. Then adding the intermediate 3f obtained by recrystallization, 10mmol hydrazine hydrate and 2mL ethanol into a reaction bottle, refluxing and stirring the reaction at 80 ℃ for 4 hours, adding water to quench the reaction after the reaction is finished, adding 5mL ethyl acetate to dilute the reaction, washing the reaction with 5mL saturated saline, drying the organic phase by anhydrous magnesium sulfate, performing spin drying, and recrystallizing by using a dichloromethane/n-hexane system to obtain a product 5f (IC)50115 μ M) as a white solid 58 mg.
1H NMR(400MHz,CDCl3)δ8.40(d,J＝8.0Hz,1H),8.11(s,1H),7.73(d,J＝6.8Hz,1H),7.44-7.34(m,3H),7.31(t,J＝8.4Hz,2H),6.62(d,J＝7.2Hz,1H),5.83(s,2H),4.10(d,J＝4.4Hz,2H).13C NMR(100MHz,CDCl3)δ163.4,141.1,137.0,134.8(q,J＝1.3Hz),132.5,128.0,127.8,127.7,127.4(q,J＝30.7Hz),126.20(q,J＝5.6Hz),124.4(q,J＝272.1Hz),123.3,122.8,109.4,49.6(q,J＝3.3Hz)。
Example 11
Synthesis of 1- (4- (trifluoromethyl) benzyl) -1H-indazole-3-carbohydrazide 5 g:
and sequentially adding weighed 0.20mmol of compound 1a, 0.22mmol of compound 2g and 0.9mmol of potassium carbonate into a reaction bottle, then adding 2mL of acetone, carrying out reflux stirring at 70 ℃ for 12 hours, after the reaction is finished, adding water to quench the reaction, adding 5mL of ethyl acetate to dilute the reaction, washing the reaction with 5mL of saturated saline solution, drying an organic phase by anhydrous magnesium sulfate, carrying out spin drying, and then recrystallizing by using a dichloromethane/n-hexane system to obtain 3g of an intermediate. Subsequently, 3g of intermediate obtained by recrystallization, 10mmol of waterHydrazine and 2mL of ethanol/water are added into a reaction bottle, the reaction is refluxed and stirred for 4 hours at the temperature of 80 ℃, after the reaction is finished, water is added for quenching the reaction, 5mL of ethyl acetate is added for dilution, 5mL of saturated saline solution is used for washing, an organic phase is dried by anhydrous magnesium sulfate, and is recrystallized by a dichloromethane/normal hexane system after being dried in a spinning mode to obtain 5g of a product (IC) which is a product50168 μ M) was 51mg of white solid.
1H NMR(400MHz,CDCl3)δ8.38(d,J＝8.4Hz,1H),8.10(s,1H),7.57(d,J＝8.0Hz,2H),7.45-7.39(m,1H),7.37-7.29(m,2H),7.28(s,2H),5.65(s,2H),4.09(d,J＝4.4Hz,2H).13C NMR(100MHz,CDCl3)δ163.4,140.9,139.9,136.9,130.6(q,J＝32.5Hz),127.6,127.5,126.0(q,J＝3.8Hz),124.0(q,J＝270.6Hz),123.4,123.3,122.9,109.4,53.0。
Example 12
(Synthesis of 1- (2, 4-dichlorobenzyl) -1H-indazole-4-carbohydrazide 5H:
and sequentially adding weighed 0.20mmol of compound 1a, 0.22mmol of compound 2h and 0.9mmol of potassium carbonate into a reaction bottle, then adding 2mL of tetrahydrofuran, carrying out reflux stirring at 70 ℃ for 18 hours, after the reaction is finished, adding water to quench the reaction, adding 5mL of ethyl acetate to dilute the reaction, washing the reaction with 5mL of saturated saline solution, drying an organic phase by anhydrous magnesium sulfate, carrying out spin drying, and then recrystallizing by using a dichloromethane/n-hexane system to obtain an intermediate for 3 hours. Then adding the intermediate obtained by recrystallization for 3h, 10mmol hydrazine hydrate and 2mL ethanol into a reaction bottle, refluxing and stirring the reaction at 80 ℃ for 4h, adding water to quench the reaction after the reaction is finished, adding 5mL ethyl acetate to dilute the reaction, washing the reaction with 5mL saturated saline, drying the organic phase with anhydrous magnesium sulfate, performing spin drying, and recrystallizing with a dichloromethane/n-hexane system to obtain a product 5h (IC)50198 μ M) as a white solid, 55 mg.
1H NMR(400MHz,CDCl3)δ8.49(s,1H),7.53(d,J＝8.0Hz,1H),7.49-7.44(m,2H),7.44-7.38(m,2H),7.10(dd,J＝8.4,2.0Hz,1H),6.70(d,J＝8.4Hz,1H),5.70(s,2H),4.20(s,2H).13C NMR(100MHz,DMSO)δ165.9,140.2,134.2,134.0,133.3,133.2,130.9,129.1,127.7,126.7,126.1,121.6,119.9,112.7,49.2。
The invention provides a preparation method of a novel estrogen receptor targeting inhibitor, wherein the reaction route of a compound 8 in the following examples is as follows:
the first step of reaction: sequentially adding weighed 2H-indazole-5-carboxylic acid methyl ester (6), alkali and 2, 4-dichloro-1- (chloromethyl) benzene (2a) into a reaction bottle, reacting at 60-80 ℃, stirring for 6-24 hours, adding water to quench the reaction after the reaction is finished, then adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, drying by spinning, and performing column chromatography separation and purification to obtain a target compound 7;
specifically, the alkali is one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate and dipotassium hydrogen phosphate, and the dosage range of the alkali is as follows: 200 and 500mol percent;
specifically, the ratio of the used amount of the base to the used amount of the compound (1) is as follows: 2:1-5:1, the amount of the base substance is 2-5 times of the amount of the compound (1) substance;
specifically, the molar part ratio of the compound (1) to the compound (2) is as follows: 1:2-1: 1;
specifically, the solvent comprises acetone, cyclohexanone, acetonitrile, tetrahydrofuran, and 1, 4-epoxy hexacyclic ring;
specifically, the reaction temperature is 60-80 ℃;
specifically, the reaction time is any time within 6 hours to 24 hours;
the second step of reaction: and sequentially adding the weighed compound 7, hydrazine hydrate and a solvent into a reaction bottle, reacting at 60-120 ℃, stirring for 12-36 hours, adding water to quench the reaction after the reaction is finished, adding ethyl acetate, filtering the solid by using a suction filter funnel, separating an organic phase by using a separating funnel, combining the organic phases, drying by spinning, and performing column chromatography separation and purification to obtain the target compound 8.
Specifically, the alkali is hydrazine hydrate, and the dosage range of the hydrazine hydrate is as follows: 100-200 mol%;
specifically, the ratio of the usage amount of the hydrazine hydrate to the usage amount of the compound (7) is as follows: 1:1-2: 1;
specifically, the solvent includes methanol, ethanol, isopropanol;
specifically, the reaction temperature is any temperature between 60 ℃ and 120 ℃;
in particular, the reaction time is any time between 2 and 6 hours.
Example 13
Synthesis of 2- (2, 4-dichlorobenzyl) -2H-indazole-5-carbohydrazide 8:
and sequentially adding weighed 0.20mmol of compound 6, 0.22mmol of 2a and 0.9mmol of potassium carbonate into a reaction bottle, then adding 2mL of acetone, carrying out reflux stirring at 70 ℃ for 12 hours, after the reaction is finished, adding water to quench the reaction, adding 5mL of ethyl acetate to dilute the reaction, washing with 5mL of saturated saline solution, drying an organic phase by anhydrous magnesium sulfate, carrying out spin drying, and then recrystallizing by using a dichloromethane/n-hexane system to obtain an intermediate 7. Then adding the intermediate 7 obtained by recrystallization, 10mmol hydrazine hydrate and 2mL ethanol into a reaction bottle, refluxing and stirring the reaction at 80 ℃ for 4 hours, adding water to quench the reaction after the reaction is finished, adding 5mL ethyl acetate to dilute the reaction, washing the reaction with 5mL saturated saline, drying an organic phase by anhydrous magnesium sulfate, performing spin drying, and recrystallizing by using a dichloromethane/n-hexane system to obtain a product 8 (IC)50119 μ M) as a white solid 53 mg.
1H NMR(400MHz,DMSO)δ9.72(s,1H),8.66(s,1H),8.28(s,1H),7.68(dd,J＝8.8,1.6Hz,2H),7.61(d,J＝8.8Hz,1H),7.44(dd,J＝8.4,2.0Hz,1H),7.17(d,J＝8.4Hz,1H),5.77(s,2H),4.47(s,2H).13C NMR(100MHz,DMSO)δ166.6,149.1,133.8,133.7,133.2,132.0,129.1,127.9,127.1,126.6,124.5,121.3,120.7,116.9,53.6。
The invention provides a preparation method of a novel estrogen receptor targeting inhibitor and application thereof in breast cancer treatment, wherein the reaction route of a compound 9 in the following embodiment is as follows:
the first step of reaction: sequentially adding weighed 1- (2, 4-dichlorobenzyl) -1H-indazole-3-carboxylic acid (4a) and a solvent into a reaction bottle, then carrying out inert gas protection, slowly dropwise adding a chlorination reagent at 0 ℃, reacting in an inert gas atmosphere, stirring for 1-3 hours at room temperature to 50 ℃, adding water to quench reaction after the reaction is finished, then adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, and directly carrying out the next reaction after spin-drying;
specifically, the chlorination reagent is one of thionyl chloride and oxalyl chloride, and the dosage range of the chlorination reagent is as follows: 110-200 mol%;
specifically, the dosage of the chlorinating agent and the material ratio of the compound (4a) are as follows: 1.1:1-2:1, the amount of the substance of the chlorinating reagent is 1.1-2 times of the amount of the substance of the compound (1);
specifically, the solvent includes tetrahydrofuran, 1, 4-dioxane, acetonitrile, and acetone;
specifically, the reaction is carried out in an inert gas atmosphere, and inert gases comprise nitrogen, helium and argon;
specifically, the reaction temperature is any one of room temperature to 50 ℃;
specifically, the reaction time is any time within 6 hours to 24 hours;
the second step of reaction: and (3) sequentially adding the crude product and ammonia water in the last step into a reaction bottle, reacting in an inert gas atmosphere, stirring at room temperature for 10-30 minutes, adding water to quench the reaction after the reaction is finished, then adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, performing column chromatography separation and purification after spin drying to obtain the target compound 9.
Specifically, the amination reagent is ammonia water, and the dosage range of the ammonia water is as follows: 100-200 mol%;
specifically, the ratio of the amount of the ammonia water to the amount of the compound (4a) is as follows: 1:1-2: 1;
specifically, the reaction temperature is room temperature;
specifically, the reaction time is any time between 10 and 30 minutes;
example 14
Synthesis of 1- (2, 4-dichlorobenzyl) -1H-indazole-3-carboxamide 9:
after weighed 1- (2, 4-dichlorobenzyl) -1H-indazole-3-carboxylic acid (4a) and tetrahydrofuran are sequentially added into a reaction bottle, inert gas protection is carried out, thionyl chloride is slowly dripped at 0 ℃, the reaction is stirred for 2 hours at 50 ℃ in an inert gas atmosphere, water is added after the reaction is finished, the reaction is quenched, then ethyl acetate is added, organic phases are separated by a separating funnel, the organic phases are combined, and an intermediate obtained after spin drying is directly used for the next reaction.
Adding the intermediate obtained in the last step and ammonia water into a reaction bottle, stirring for 10 minutes at room temperature, adding water to quench the reaction after the reaction is finished, adding 5mL of ethyl acetate to dilute the reaction solution, washing with 5mL of saturated saline solution, drying the organic phase by anhydrous magnesium sulfate, and recrystallizing by a dichloromethane/n-hexane system after spin-drying to obtain a product 9 (IC)50498 μ M) as a white solid 46 mg.
1H NMR(400MHz,CDCl3)δ8.41(d,J＝8.0Hz,1H),7.47-7.44(m,1H),7.44-7.40(m,1H),7.39-7.35(m,1H),7.33(t,J＝7.4Hz,1H),7.12(dd,J＝8.4,2.0Hz,1H),6.88(s,1H),6.67(d,J＝8.4Hz,1H),5.68(s,2H),5.51(s,1H).13C NMR(100MHz,CDCl3)δ164.5,141.3,137.9,134.7,133.3,132.4,129.62,129.55,127.8,127.6,123.32,123.26,123.2,109.4,50.2.。
The invention provides a preparation method of a novel estrogen receptor targeting inhibitor and application thereof in breast cancer treatment, wherein the reaction route of a compound 13 in the following embodiment is as follows:
the first step of reaction: sequentially adding weighed 1H-indole-3-carboxylic acid methyl ester (10), alkali and 2, 4-dichloro-1- (chloromethyl) benzene (2a) into a reaction bottle, adding a reaction solvent, reacting, stirring for 6-24 hours at 60-80 ℃, adding water for quenching reaction after the reaction is finished, then adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, and performing column chromatography separation and purification after spin drying to obtain a target compound 11;
specifically, the alkali is one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate and dipotassium hydrogen phosphate, and the dosage range of the alkali is as follows: 200 and 500mol percent;
specifically, the ratio of the used amount of the base to the used amount of the compound (10) is as follows: 2:1-5:1, the amount of the base substance is 2-5 times of the amount of the compound (1) substance;
specifically, the molar ratio of the compound (110) to the compound (2a) is as follows: 1:2-1: 1;
specifically, the solvent comprises acetone, cyclohexanone, acetonitrile, tetrahydrofuran, and 1, 4-epoxy hexacyclic ring;
specifically, the reaction temperature is 60-80 ℃;
specifically, the reaction time is any time within 6 hours to 24 hours;
the second step of reaction: sequentially adding the weighed compound 11, hydrazine hydrate and solvent into a reaction bottle, then carrying out inert gas protection, stirring for 12-36 hours at the temperature of 60-120 ℃ for reaction, adding water to quench the reaction after the reaction is finished, then adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, carrying out spin drying, and carrying out column chromatography separation and purification to obtain a target compound 13;
specifically, the alkali is hydrazine hydrate, and the dosage range of the hydrazine hydrate is as follows: 100-200 mol%;
specifically, the dosage of the hydrazine hydrate and the dosage ratio of the compound (11) are as follows: 1:1-2: 1;
specifically, the solvent includes methanol, ethanol, isopropanol;
specifically, the reaction temperature is any temperature between 60 ℃ and 120 ℃;
specifically, the reaction time is any time between 2 and 6 hours;
example 15
Synthesis of (1- (2, 4-dichlorobenzyl) -1H-indole-3-carbohydrazide 13:
and sequentially adding weighed 0.20mmol of compound 10, 0.22mmol of 2a and 0.9mmol of potassium carbonate into a reaction bottle, then adding 2mL of acetone, carrying out reflux stirring at 70 ℃ for 12 hours, after the reaction is finished, adding water to quench the reaction, adding 5mL of ethyl acetate to dilute the reaction, washing 5mL of saturated saline solution, drying an organic phase by anhydrous magnesium sulfate, carrying out spin drying, and then recrystallizing by a dichloromethane/n-hexane system to obtain an intermediate 11. Then adding intermediate 11 obtained by recrystallization, 10mmol hydrazine hydrate and 2mL methanol into a reaction bottle, refluxing and stirring the reaction at 80 ℃ for 4 hours, adding water to quench the reaction after the reaction is finished, adding 5mL ethyl acetate to dilute the reaction, washing the reaction with 5mL saturated saline, drying an organic phase by anhydrous magnesium sulfate, performing spin drying, and recrystallizing by using a dichloromethane/n-hexane system to obtain a product 13 (IC)50237 μ M) as a white solid 53 mg.
1H NMR(400MHz,CDCl3)δ7.96(d,J＝6.0Hz,1H),7.71(s,1H),7.46(d,J＝2.0Hz,1H),7.33-7.27(m,3H),7.17(s,1H),7.11(dd,J＝8.4,2.0Hz,1H),6.63(d,J＝8.4Hz,1H),5.39(s,2H),4.15(s,2H).13C NMR(100MHz,DMSO)δ164.7,136.2,133.6,133.5,130.8,130.5,129.2,127.9,126.7,122.5,121.5,121.1,110.5,109.1,47.0.
The invention provides a preparation method of a novel estrogen receptor targeting inhibitor and application thereof in breast cancer treatment, wherein the reaction route of a compound 14 in the following embodiment is as follows:
the first step of reaction: sequentially adding weighed 1H-indole-3-carboxylic acid methyl ester (10), alkali and 2, 4-dichloro-1- (chloromethyl) benzene (2a) into a reaction bottle, then adding a reaction solvent, reacting, stirring for 6-24 hours at 60-80 ℃, adding water for quenching reaction after the reaction is finished, then adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, and performing column chromatography separation and purification after spin drying to obtain a target compound 11;
specifically, the alkali is one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate and dipotassium hydrogen phosphate, and the dosage range of the alkali is as follows: 200 and 500mol percent;
specifically, the ratio of the used amount of the base to the used amount of the compound (10) is as follows: 2:1-5:1, the amount of the base substance is 2-5 times of the amount of the compound (1) substance;
specifically, the molar ratio of the compound (110) to the compound (2a) is as follows: 1:2-1: 1;
specifically, the solvent comprises acetone, cyclohexanone, acetonitrile, tetrahydrofuran, and 1, 4-epoxy hexacyclic ring;
specifically, the reaction temperature is 60-80 ℃;
specifically, the reaction time is any time within 6 hours to 24 hours;
the second step of reaction: sequentially adding the weighed compound 11, alkali and solvent into a reaction bottle, reacting at 60-120 ℃ for 12-36 hours in an inert gas atmosphere, adding water to quench the reaction after the reaction is finished, adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, and performing column chromatography separation and purification to obtain a target compound 12 after the organic phases are dried in a spinning mode;
specifically, the alkali is one of sodium hydroxide and potassium hydroxide, and the dosage range of the alkali is as follows: 100-200 mol%;
specifically, the ratio of the used amount of the base to the used amount of the compound (1) is as follows: 1:1-2:1, the amount of the base substance being 1-2 times the amount of the compound (1) substance;
specifically, the solvent includes methanol/water, ethanol/water, tetrahydrofuran/water, acetone/water;
specifically, the reaction temperature is any temperature between room temperature and 60 ℃;
the temperature is any temperature between room temperature and 60 ℃;
specifically, the reaction time is any time between 12 and 36 hours;
the third step of reaction: and sequentially adding the weighed compound 12 and a solvent into a reaction bottle, then carrying out inert gas protection, slowly dropwise adding a chlorination reagent at 0 ℃, reacting in an inert gas atmosphere at room temperature for 1-3 hours, adding water to quench the reaction after the reaction is finished, then adding ethyl acetate, separating an organic phase by using a separating funnel, combining the organic phases, and directly carrying out the next reaction after spin drying.
Specifically, the chlorination reagent is one of thionyl chloride and oxalyl chloride, and the dosage range of the chlorination reagent is as follows: 110-200 mol%;
specifically, the dosage of the chlorinating agent and the material ratio of the compound (4a) are as follows: 1.1:1-2:1, the amount of the substance of the chlorinating reagent is 1.1-2 times of the amount of the substance of the compound (1);
specifically, the solvent includes tetrahydrofuran, 1, 4-dioxane, acetonitrile, and acetone;
specifically, the reaction is carried out in an inert gas atmosphere, and inert gases comprise nitrogen, helium and argon;
specifically, the reaction temperature is any one of room temperature to 50 ℃;
specifically, the reaction time is any time within 2 hours to 6 hours;
and a fourth step of reaction: and (3) sequentially adding the crude product and ammonia water in the last step into a reaction bottle, reacting in an inert gas atmosphere, stirring at room temperature for 10-30 minutes, adding water to quench the reaction after the reaction is finished, then adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, performing column chromatography separation and purification after spin drying to obtain the target compound 14.
Specifically, the alkali is hydrazine hydrate, and the dosage range of the hydrazine hydrate is as follows: 100-200 mol%;
specifically, the dosage of the hydrazine hydrate and the dosage ratio of the compound (12) are as follows: 1:1-2: 1;
specifically, the solvent includes methanol, ethanol, isopropanol;
specifically, the reaction temperature is room temperature;
in particular, the reaction time is any time between 10 and 30 minutes.
Example 16
Synthesis of 1- (2, 4-dichlorobenzyl) -1H-indole-3-carboxamide 14:
and sequentially adding weighed 0.20mmol of compound 10, 0.22mmol of 2a and 0.9mmol of potassium carbonate into a reaction bottle, then adding 2mL of acetone, carrying out reflux stirring at 70 ℃ for 12 hours, after the reaction is finished, adding water to quench the reaction, adding 5mL of ethyl acetate to dilute the reaction, washing 5mL of saturated saline solution, drying an organic phase by anhydrous magnesium sulfate, carrying out spin drying, and then recrystallizing by a dichloromethane/n-hexane system to obtain an intermediate 11.
Subsequently, the intermediate 11 obtained by recrystallization, 0.40mmol of sodium hydroxide and 2mL of tetrahydrofuran/water (1:1) mixed solvent are added into a reaction bottle, the reaction is stirred at room temperature for 24 hours, after the reaction is finished, water is added to quench the reaction, 5mL of ethyl acetate is added for dilution, 5mL of saturated saline solution is used for washing, the organic phase is dried by anhydrous magnesium sulfate, and after the drying, the dichloromethane/normal hexane system is used for recrystallization to obtain a white solid product 12 of 50 mg.
Adding the weighed compound 12 and tetrahydrofuran in turn into a reaction bottle, then carrying out inert gas protection, slowly dropwise adding thionyl chloride at 0 ℃, reacting in an inert gas atmosphere, stirring at 50 ℃ for 2 hours, adding water after the reaction is finished, quenching the reaction, then adding ethyl acetate, separating an organic phase by using a separating funnel, combining the organic phases, and spin-drying to obtain an intermediate which is directly used for the next reaction.
The intermediate obtained in the previous step is hydrated by 10mmolAdding hydrazine and 2mL of ethanol into a reaction bottle, stirring for 4 hours at room temperature, adding water to quench the reaction after the reaction is finished, adding 5mL of ethyl acetate to dilute the reaction solution, washing the reaction solution with 5mL of saturated saline solution, drying an organic phase by using anhydrous magnesium sulfate, performing spin drying, and recrystallizing by using a dichloromethane/normal hexane system to obtain a product 14 (IC)50482 μ M) as a white solid 40 mg.
1H NMR(400MHz,CDCl3)δ8.01(d,J＝6.8Hz,1H),7.73(s,1H),7.46(d,J＝2.0Hz,1H),7.35-7.27(m,3H),7.11(dd,J＝8.0,2.0Hz,1H),6.64(d,J＝8.4Hz,1H),5.73(s,2H),5.39(s,2H).13C NMR(100MHz,DMSO)δ166.2,136.4,133.7,133.43,133.39,131.8,130.7,129.2,127.9,126.8,122.5,121.6,121.1,110.6,110.5,47.0.
The invention provides a preparation method of a novel estrogen receptor targeting inhibitor and application thereof in breast cancer treatment, wherein the reaction route of a compound 19 in the following embodiment is as follows:
the first step of reaction: adding the weighed indazole (15), iodination reagent and solvent into a reaction bottle in sequence, reacting, stirring at room temperature for 2-4 hours, adding water to quench the reaction after the reaction is finished, adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, performing column chromatography separation and purification after spin drying to obtain a target compound 16;
specifically, the alkali is one of sodium hydroxide and potassium hydroxide, and the dosage range of the alkali is as follows: 100-200 mol%;
specifically, the ratio of the used amount of the base to the used amount of the compound (15) is as follows: 1:1-2:1, the amount of the base substance being 1-2 times the amount of the compound (1) substance;
specifically, the compound (15) is reacted with iodine I2The molar ratio ranges are as follows: 1:2-1: 1;
specifically, the solvent is N, N-dimethylformamide;
specifically, the reaction temperature is room temperature;
specifically, the reaction time is any time within 2 to 4 hours;
the second step of reaction: sequentially adding the weighed compound 16, alkali and 2, 4-dichloro-1- (chloromethyl) benzene (2a) into a reaction bottle, then adding a reaction solvent, reacting at 60-80 ℃, stirring for 6-24 hours, after the reaction is finished, adding water to quench the reaction, then adding ethyl acetate, separating an organic phase by using a separating funnel, combining the organic phases, and after spin-drying, performing column chromatography separation and purification to obtain a target compound 17;
specifically, the alkali is one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate and dipotassium hydrogen phosphate, and the dosage range of the alkali is as follows: 200 and 500mol percent;
specifically, the ratio of the used amount of the base to the used amount of the compound (10) is as follows: 2:1-5:1, the amount of the base substance is 2-5 times of the amount of the compound (1) substance;
specifically, the molar ratio of the compound (110) to the compound (2a) is as follows: 1:2-1: 1;
specifically, the solvent comprises acetone, cyclohexanone, acetonitrile, tetrahydrofuran, and 1, 4-epoxy hexacyclic ring;
specifically, the reaction temperature is 60-80 ℃;
specifically, the reaction time is any time within 6 hours to 24 hours;
the third step of reaction: adding the weighed compound 17, a palladium catalyst, alkali, a phosphine ligand and allyl formate into a reaction bottle in sequence, then adding a reaction solvent, then carrying out inert gas protection, stirring for 12-24 hours at the temperature of 100-120 ℃ for reaction, adding water to quench the reaction after the reaction is finished, then adding ethyl acetate, filtering the solid by using a suction filter funnel, then separating the organic phase by using a separating funnel, combining the organic phases, carrying out spin drying, and carrying out column chromatography separation and purification to obtain the target compound 18.
Specifically, the metal palladium catalyst is one of palladium acetate, palladium chloride, palladium bromide and palladium trifluoroacetate, and the dosage range of the catalyst is as follows: 10-20 mol%;
specifically, the phosphine ligand is one of triphenylphosphine, tricyclohexylphosphine and 1, 2-bis (diphenylphosphino) ethane, and the dosage range of the phosphine ligand is as follows: 25-50 mol%;
specifically, the base is one of N, N-diisopropylethylamine and triethylamine, and the dosage range of the base is as follows: 200-300 mol%;
specifically, the molar ratio of the compound (17) to the allyl formate is as follows: 1:2-1: 5;
specifically, the reaction solvent is N, N-dimethylformamide;
specifically, the reaction is carried out in an inert gas atmosphere, and inert gases comprise nitrogen, helium and argon;
specifically, the temperature in the reaction is any one of 100 ℃ to 120 ℃,
in particular, the reaction time is any time between 12 and 24 hours.
And a fourth step of reaction: and sequentially adding the weighed compound 18, hydrazine hydrate and a solvent into a reaction bottle, then carrying out inert gas protection, stirring for 2-6 hours at the temperature of 60-120 ℃ for reaction, adding water to quench the reaction after the reaction is finished, then adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, carrying out spin drying, and carrying out column chromatography separation and purification to obtain the target compound 19.
Specifically, the alkali is hydrazine hydrate, and the dosage range of the hydrazine hydrate is as follows: 100-200 mol%;
specifically, the dosage of the hydrazine hydrate and the dosage ratio of the compound (18) are as follows: 1:1-2: 1;
specifically, the solvent includes methanol, ethanol, isopropanol;
specifically, the reaction temperature is any temperature between 60 ℃ and 120 ℃;
in particular, the reaction time is any time between 2 and 6 hours.
Example 17
Synthesis of 3- (1- (2, 4-dichlorobenzyl) -1H-indazol-3-yl) propionylhydrazine 19:
the weighed compound 15 in 0.20mmol and iodine I in 0.40mmol are added into a reaction bottle in sequence2And 2mL of DMF (dimethyl formamide), stirring at room temperature for 2 hours, after the reaction is finished, adding water to quench the reaction, adding 5mL of ethyl acetate to dilute the reaction, washing the reaction with 5mL of saturated saline solution, drying an organic phase by anhydrous magnesium sulfate, and spin-drying to obtain an intermediate 16 which is directly used for the next reaction.
And sequentially adding weighed 0.20mmol of compound 16, 0.22mmol of compound 2a and 0.9mmol of potassium carbonate into a reaction bottle, then adding 2mL of acetone, carrying out reflux stirring at 70 ℃ for 12 hours, after the reaction is finished, adding water to quench the reaction, adding 5mL of ethyl acetate to dilute the reaction, washing the reaction with 5mL of saturated saline solution, drying an organic phase by anhydrous magnesium sulfate, carrying out spin drying, and then recrystallizing by using a dichloromethane/n-hexane system to obtain an intermediate 17.
And sequentially adding weighed 0.20mmol of compound 17, 0.02mmol of palladium acetate, 0.60mmol of DIPEA, 0.1mmol of triphenylphosphine and 2Ml of DMF into a reaction bottle, then carrying out inert gas protection, stirring the mixture at 120 ℃ for 24 hours, adding water to quench the reaction after the reaction is finished, then adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, carrying out spin drying, and recrystallizing by using a dichloromethane/n-hexane system to obtain an intermediate 18.
Adding 0.20mmol of intermediate 18 obtained in the previous step, 10mmol of hydrazine hydrate and 2mL of ethanol into a reaction bottle, stirring for 4 hours at room temperature, adding water to quench the reaction after the reaction is finished, adding 5mL of ethyl acetate to dilute the reaction, washing with 5mL of saturated saline, drying an organic phase with anhydrous magnesium sulfate, performing spin drying, and recrystallizing with a dichloromethane/n-hexane system to obtain a product 19 (IC)50145 μ M) was 55mg of white solid.
1H NMR(400MHz,CDCl3)δ7.71(d,J＝8.0Hz,1H),7.42(d,J＝2.0Hz,1H),7.40-7.34(m,1H),7.32-7.27(m,1H),7.21-7.12(m,2H),7.09(dd,J＝8.4,2.0Hz,1H),6.60(d,J＝8.4Hz,1H),5.59(s,2H),3.83(s,2H),3.34(t,J＝7.2Hz,2H),2.73(t,J＝7.2Hz,2H).13C NMR(100MHz,CDCl3)δ173.4,144.9,140.9,134.1,133.5,133.1,129.4,129.4,127.6,127.2,123.0,120.7,120.5,109.2,49.4,33.2,22.7.
The invention provides a preparation method of a novel estrogen receptor targeting inhibitor and application thereof in breast cancer treatment, wherein the reaction route of a compound 21 in the following embodiment is as follows:
the first step of reaction: sequentially adding the weighed compound (17), a palladium catalyst, alkali and (4- (methoxycarbonyl) phenyl) boric acid into a reaction bottle, then adding a reaction solvent, then carrying out inert gas protection, reacting at 60-80 ℃ in an inert gas atmosphere, stirring for 12-24 hours, after the reaction is finished, adding water to quench the reaction, then adding ethyl acetate, separating an organic phase by using a separating funnel, combining the organic phases, carrying out spin drying, and carrying out column chromatography separation and purification to obtain a target compound 20;
specifically, the metal palladium catalyst is one of 1,1' -bis-diphenylphosphino ferrocene palladium dichloride, palladium acetate, palladium chloride, palladium bromide and palladium trifluoroacetate, and the dosage range of the catalyst is as follows: 10-20 mol%;
specifically, the alkali is one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate and dipotassium hydrogen phosphate, and the dosage range of the alkali is as follows: 100-200 mol%;
specifically, the ratio of the amount of the base to the amount of the compound (17) is as follows: 1:1-2:1, the amount of the base substance being 1-2 times the amount of the compound (1) substance;
specifically, the molar ratio of the compound (17) to the (4- (methoxycarbonyl) phenyl) boric acid is in the range of: 1:2-1: 1;
specifically, the solvent is a mixed solvent of ethanol/toluene/water;
specifically, the reaction is carried out in an inert gas atmosphere, and inert gases comprise nitrogen, helium and argon;
specifically, the reaction temperature is any one of 60 ℃ to 80 ℃;
specifically, the reaction time is any time within 12 hours to 24 hours;
the second step of reaction: and sequentially adding the weighed compound 20, hydrazine hydrate and solvent into a reaction bottle, reacting at 60-120 ℃ for 2-6 hours under stirring, adding water to quench the reaction after the reaction is finished, adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, performing column chromatography separation and purification after spin drying to obtain the target compound 21.
Specifically, the alkali is hydrazine hydrate, and the dosage range of the hydrazine hydrate is as follows: 100-200 mol%;
specifically, the dosage of the hydrazine hydrate and the dosage ratio of the compound (20) are as follows: 1:1-2: 1;
specifically, the solvent includes methanol, ethanol, isopropanol;
specifically, the reaction temperature is any temperature between 60 ℃ and 120 ℃;
specifically, the reaction time is any time between 2 and 6 hours;
example 18
Synthesis of 4- (1- (2, 4-dichlorobenzyl) -1H-indazol-3-yl) benzoyl hydrazine 21:
the weighed compound 17 in 0.20mmol and Pd (dppf) Cl in 0.02mmol are sequentially added into a reaction bottle2After 0.40mmol of sodium carbonate, 0.30mmol of (4- (methoxycarbonyl) phenyl) boric acid and 2mL of ethanol/acetone/water (1:1:1) mixed solvent are subjected to inert gas protection, the reaction is stirred for 24 hours at 80 ℃, water is added after the reaction is finished to quench the reaction, then ethyl acetate is added, organic phases are separated by a separating funnel, the organic phases are combined, and after spin drying, the mixture is recrystallized by a dichloromethane/n-hexane system to obtain an intermediate 20.
Adding 0.20mmol of intermediate 20 obtained in the previous step, 10mmol of hydrazine hydrate and 2mL of ethanol into a reaction bottle, stirring for 4 hours at room temperature, adding water to quench the reaction after the reaction is finished, adding 5mL of ethyl acetate to dilute the reaction, washing with 5mL of saturated saline solution, drying an organic phase with anhydrous magnesium sulfate, performing spin drying, and recrystallizing with a dichloromethane/n-hexane system to obtain a product 21(IC5045 μ M) as a white solid 55 mg.
1H NMR(400MHz,CDCl3)δ8.09(d,J＝8.4Hz,2H),8.06(d,J＝8.4Hz,1H),7.89(d,J＝8.8Hz,2H),7.44(d,J＝2.0Hz,2H),7.43-7.37(m,2H),7.32-7.27(m,1H),7.09(dd,J＝8.4,2.0Hz,1H),6.74(d,J＝8.4Hz,1H),5.73(s,2H),4.15(s,2H).13C NMR(100MHz,DMSO)δ165.7,142.6,141.4,135.6,133.9,133.3,133.2,132.6,130.8,129.1,127.81,127.78,127.0,126.7,122.1,121.2,121.0,110.4,49.3.
The invention provides a preparation method of a novel estrogen receptor targeting inhibitor and application thereof in breast cancer treatment, wherein the reaction route of a compound 23 in the following embodiment is as follows:
the first step of reaction: sequentially adding the weighed compound (17), a palladium catalyst, alkali and (3- (methoxycarbonyl) phenyl) boric acid into a reaction bottle, then adding a reaction solvent, then carrying out inert gas protection, reacting at 60-80 ℃ in an inert gas atmosphere, stirring for 6-24 hours, after the reaction is finished, adding water to quench the reaction, then adding ethyl acetate, separating an organic phase by using a separating funnel, combining the organic phases, carrying out spin drying, and carrying out column chromatography separation and purification to obtain a target compound 22;
specifically, the metal palladium catalyst is one of 1,1' -bis-diphenylphosphino ferrocene palladium dichloride, palladium acetate, palladium chloride, palladium bromide and palladium trifluoroacetate, and the dosage range of the catalyst is as follows: 10-20 mol%;
specifically, the alkali is one of potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, potassium phosphate, potassium hydrogen phosphate and dipotassium hydrogen phosphate, and the dosage range of the alkali is as follows: 100-200 mol%;
specifically, the ratio of the amount of the base to the amount of the compound (17) is as follows: 1:1-2:1, the amount of the base substance being 1-2 times the amount of the compound (1) substance;
specifically, the molar ratio of the compound (17) to the (3- (methoxycarbonyl) phenyl) boric acid is in the range of: 1:2-1: 1;
specifically, the solvent is a mixed solvent of ethanol/toluene/water;
specifically, the reaction is carried out in an inert gas atmosphere, and the inert gas comprises nitrogen, helium and argon.
Specifically, the reaction temperature is any one of 60 ℃ to 80 ℃;
specifically, the reaction time is any time within 12 hours to 24 hours;
the second step of reaction: and sequentially adding the weighed compound 22, hydrazine hydrate and solvent into a reaction bottle, reacting at 60-120 ℃ for 2-6 hours under stirring, adding water to quench the reaction after the reaction is finished, adding ethyl acetate, separating organic phases by using a separating funnel, combining the organic phases, performing column chromatography separation and purification after spin drying to obtain the target compound 23.
Specifically, the alkali is hydrazine hydrate, and the dosage range of the hydrazine hydrate is as follows: 100-200 mol%;
specifically, the ratio of the usage amount of the hydrazine hydrate to the usage amount of the compound (22) is as follows: 1:1-2:1,
specifically, the solvent includes methanol, ethanol, isopropanol;
specifically, the reaction temperature is any temperature between 60 ℃ and 120 ℃;
specifically, the reaction time is any time between 2 and 6 hours;
example 19
Synthesis of 3- (1- (2, 4-dichlorobenzyl) -1H-indazol-3-yl) benzoyl hydrazine 23:
the weighed compound 17 in 0.20mmol and Pd (dppf) Cl in 0.02mmol are sequentially added into a reaction bottle20.40mmol of sodium carbonate, 0.30mmol of (4- (methoxycarbonyl) phenyl) boric acid and 2mL of ethanol/acetone/water (1:1:1) mixed solvent, then carrying out inert gas protection, stirring the reaction at 80 ℃ for 24 hours, adding water to quench the reaction after the reaction is finished, and then adding water to quench the reactionEthyl acetate, separating the organic phases by using a separating funnel, combining the organic phases, carrying out spin drying, and then recrystallizing by using a dichloromethane/n-hexane system to obtain an intermediate 22.
Adding 0.20mmol of intermediate 22 obtained in the previous step, 10mmol of hydrazine hydrate and 2mL of ethanol into a reaction bottle, stirring for 4 hours at room temperature, adding water to quench the reaction after the reaction is finished, adding 5mL of ethyl acetate to dilute the reaction, washing with 5mL of saturated saline, drying an organic phase with anhydrous magnesium sulfate, performing spin drying, and recrystallizing with a dichloromethane/n-hexane system to obtain a product 23 (IC)5053 μ M) as a white solid 58 mg.
1H NMR(400MHz,CDCl3)δ8.36(t,J＝1.6Hz,1H),8.15(dt,J＝7.6,1.2Hz,1H),8.06(d,J＝8.0Hz,1H),7.79(dt,J＝8.0,1.2Hz,1H),7.60(t,J＝7.6Hz,1H),7.50(s,1H),7.44(d,J＝2.4Hz,1H),7.43-7.37(m,2H),7.31-7.27(m,1H),7.09(dd,J＝8.4,2.4Hz,1H),6.72(d,J＝8.4Hz,1H),5.73(s,2H),4.15(s,2H).13C NMR(100MHz,DMSO)δ165.9,142.9,141.4,134.2,133.9,133.3,133.21,133.16,130.7,129.6,129.2,129.1,127.8,127.0,126.6,125.5,121.9,121.2,120.9,110.3,49.3。
The above description is only a preferred embodiment of the present invention, and should not be taken as limiting the invention, and any local variations in the formulation and process thereof should be considered within the scope of the present invention.
- 上一篇：一种医用注射器针头装配设备
- 下一篇：4,5-二氰基-2-三氟甲基咪唑盐的合成方法