阅读说明：本技术 2-甲氧基雌莫司汀及其衍生物、制备方法和应用 (2-methoxy estramustine and derivatives, preparation method and application thereof ) 是由 周亚耀 于 2020-03-02 设计创作，主要内容包括：本发明公开了2-甲氧基雌莫司汀及其衍生物,其结构式如式1～4所示。本发明所述2-甲氧基雌莫司汀以2-甲氧雌二醇为载体,将烷化剂(氮芥)通过氨基甲酸酯连接组成双功能药物分子化合物,整个分子作为抗有丝分裂剂,氨基甲酸酯在体内代谢水解后,代谢物介导释放的2-甲氧基雌二醇仍可以继续发挥抗肿瘤作用。2-甲氧基雌莫司汀的衍生物可以作为2-甲氧基雌二醇的前药。本发明还公开了2-甲氧基雌莫司汀及其衍生物的制备方法,该制备方法可以制得2-甲氧基雌莫司汀及其衍生物,且收率较高。本发明还公开了2-甲氧基雌莫司汀及其衍生物在治疗肿瘤或多发性骨髓瘤药物中的应用。(The invention discloses 2-methoxy estramustine and derivatives thereof, and the structural formula of the 2-methoxy estramustine and the derivatives thereof is shown in formulas 1-4. The 2-methoxyestramustine takes 2-methoxyestradiol as a carrier, an alkylating agent (nitrogen mustard) is connected through carbamate to form a dual-functional drug molecular compound, the whole molecule is taken as an antimitotic agent, and after the carbamate is metabolized and hydrolyzed in vivo, the metabolite-mediated released 2-methoxyestradiol can still continuously play an antitumor role. Derivatives of 2-methoxyestramustine can be used as prodrugs of 2-methoxyestradiol. The invention also discloses a preparation method of the 2-methoxy estramustine and the derivatives thereof, and the preparation method can prepare the 2-methoxy estramustine and the derivatives thereof with higher yield. The invention also discloses application of the 2-methoxy estramustine and the derivatives thereof in medicaments for treating tumors or multiple myeloma.)

1.2-methoxy estramustine and derivatives thereof, which are characterized in that the structural formula is shown as formulas 1-4:

wherein R is an amino acid residue; HY is hydrochloric acid, sulfuric acid, phosphoric acid, tartaric acid, malic acid or p-toluenesulfonic acid; n is 2 or 3.

2. The process for the preparation of 2-methoxyestramustine and derivatives thereof according to claim 1, wherein the starting materials are selected from the group consisting of,

the synthetic routes of the compounds represented by formula 1 and formula 2 are as follows:

the preparation method of the compound shown in the formula 1 comprises the following steps:

dissolving the compound shown in the formula 5 in dichloromethane, adding triethylamine and 4-dimethylaminopyridine, and uniformly stirring; dissolving N, N-bis (2-chloroethyl) carbamoyl chloride in dichloromethane, dropwise adding the dichloromethane into the reaction solution, stirring for reaction for 0.4-0.6 hour, then moving the reaction solution to a water bath with the temperature of 38-42 ℃, keeping the temperature for reaction till the reaction is complete, stopping the reaction, cooling the reaction solution to room temperature, adding distilled water, adjusting the pH of the reaction solution to be 5-6 by using a hydrochloric acid solution under stirring, extracting to obtain an organic phase, and removing the solvent by reduced pressure distillation to obtain the compound shown in the formula 1; wherein the molar ratio of the compound shown in the formula 5 to the triethylamine to the 4-dimethylaminopyridine to the N, N-bis (2-chloroethyl) carbamoyl chloride is as follows: 1 (1.8-2.2), (0.08-0.12), (1.2-1.8);

the preparation method of the compound shown in the formula 2 comprises the following steps:

(1) dropping phosphorus oxychloride into pyridine at 4-6 ℃, and uniformly stirring to obtain a pyridine solution of phosphorus oxychloride; dissolving the compound shown in the formula 1 in pyridine, dropwise adding the pyridine solution into a pyridine solution of phosphorus oxychloride, after the reaction is completed, pouring the reaction solution into an ice-water mixture, adding a hydrochloric acid solution to adjust the pH value to be 2-3, filtering, taking filter residue, drying, dissolving the filter residue in ethyl acetate, adding ethanol, dropwise adding distilled water until the generated turbidity does not disappear, adding a small amount of ethyl acetate to redissolve, continuously stirring, filtering, taking the filter residue, and drying to obtain the compound shown in the formula 6; wherein the mol ratio of the compound shown in the formula 1 to the phosphorus oxychloride is as follows: 1: 4-6

(2) Adding anhydrous methanol into the compound shown in the formula 6, dissolving, and cooling to 0-5 ℃ to obtain a methanol solution of the compound shown in the formula 6; dissolving metal sodium in absolute methanol, and cooling to 5-8 ℃ to obtain a methanol solution of sodium methoxide; dropwise adding a methanol solution of sodium methoxide into a methanol solution of a compound shown in a formula 6, stirring and reacting for 8-15 minutes at 0-5 ℃, adding anhydrous ether at 0-5 ℃, filtering, taking filter residues, washing the filter residues with the anhydrous ether, and drying to obtain a compound shown in a formula 2; wherein, the mol ratio of the compound shown in the formula 6 to the metallic sodium is as follows: 1: 1.1-1.3;

the synthetic route of the compound represented by formula 3 is as follows:

the preparation method of the compound shown in the formula 3 comprises the following steps:

dissolving the compound shown in the formula 1 in dichloromethane, adding triethylamine, stirring uniformly, slowly adding glycyl chloride hydrochloride, stirring until the reaction is complete, adding distilled water, taking an organic phase, washing with inclusion sodium chloride, drying with anhydrous sodium sulfate, distilling under reduced pressure to remove part of a solvent, dropwise adding a HY 1, 4-dioxane solution, stirring, filtering, and taking filter residues to obtain the compound shown in the formula 3; wherein the compound shown in the formula 1, triethylamine, glycinyl chloride hydrochloride and HY have the following molar ratio: 1, (2.5-3.5): (1.2-1.8): 3-5;

the synthetic route of the compound represented by formula 4 is as follows:

the preparation method of the compound shown in the formula 4 comprises the following steps:

dissolving the compound shown in the formula 1 in dichloromethane, adding triethylamine and 4-dimethylaminopyridine, and uniformly stirring; dissolving succinic anhydride or glutaric anhydride in dichloromethane, dropwise adding the succinic anhydride or glutaric anhydride into the reaction solution, stirring until the reaction is complete, stopping the reaction, cooling the reaction solution to room temperature, adding distilled water, adjusting the pH of the reaction solution to be 5-6 by using a hydrochloric acid solution under stirring, extracting to obtain an organic phase, washing, removing the solvent to obtain a solid, dissolving the solid in methanol and water, adding sodium bicarbonate, stirring, distilling under reduced pressure to remove the solvent to obtain a solid, and recrystallizing ethanol and diethyl ether to obtain a compound shown in a formula 4; wherein, the mole ratio of the compound shown in the formula 1, triethylamine, 4-dimethylamino pyridine and succinic anhydride or glutaric anhydride is as follows: 1: (1.8-2.2): (0.08-0.12): (1-1.2).

3. The use of 2-methoxyestramustine or a derivative thereof according to claim 1 for the preparation of an anti-tumor medicament.

4. The use of claim 3, wherein the tumor is prostate cancer, breast cancer or colon cancer.

5. The use of 2-methoxyestramustine or a derivative thereof according to claim 1 for the preparation of a medicament for the treatment of multiple myeloma.

Technical Field

The invention relates to the technical field of organic synthesis, in particular to a novel compound 2-methoxy estramustine and derivatives, a preparation method and application thereof.

Background

At present, with the improvement of medical level, the tumor treatment also makes great progress, the life cycle of tumor patients is obviously prolonged, the life quality is obviously improved, and the invention and the clinical application of various high-efficiency anticancer and antitumor drugs are mainly benefited. However, some tumor drugs used in clinical practice at present are accompanied by some side effects and adverse reactions while achieving significant curative effects, thereby causing great pain to patients and seriously affecting the quality of life of patients.

At present, the development and research on nitrogen mustard antineoplastic compounds are focused on modifying the structural part of a carrier, mainly through designing nitrogen mustard drugs with targeting carriers to reduce the toxic and side effects, estramustine sodium phosphate is also called estramustine phosphate and is clinically used for treating prostatic cancer, estramustine phosphate is taken as a carrier, nitrogen mustard (an alkylating agent) and estradiol-17- β -phosphate (hormone) are connected through carbamate, the whole molecule of the estramustine phosphate is an antimitotic agent, after the carbamate is hydrolyzed, estrogen released by mediation of metabolites plays a role of anti-gonadotropin, estramustine can specifically introduce the drugs into prostate tissues through steroid receptors to generate cytotoxic effect and hormone effect, and the purpose of damaging cancer cells is achieved.

Tumor growth and metastasis depend on the nutrition and oxygen supply of blood vessels, so cutting off tumor angiogenesis is one of the main directions for seeking antitumor drugs at present. 2-methoxyestradiol (5) is a neovascular inhibitor which enters phase I-III clinical research and is proved to be effective in treating various solid tumors, particularly breast cancer, prostatic cancer, multiple myeloma and the like. Because 2-methoxyestradiol is an endogenous active metabolite of a human body and shows low toxicity clinically, the development prospect of 2-methoxyestradiol as a novel tumor chemotherapeutic drug has great advantages.

Disclosure of Invention

The invention aims to overcome the defects of the prior art and provides 2-methoxy estramustine and derivatives, a preparation method and application thereof.

In order to achieve the purpose, the invention adopts the technical scheme that:

the structural formula of the 2-methoxy estramustine and the derivatives thereof is shown as the formula 1-4:

wherein R is amino acid residue, can be L-amino acid, such as glycine, alanine, histidine and other common amino acids, HY is hydrochloric acid, sulfuric acid, phosphoric acid, tartaric acid, malic acid or p-toluenesulfonic acid, and n is 2 or 3, when n is 2 or 3, the corresponding organic dibasic acid sodium salt is different, such as succinic acid, glutaric acid.

The invention also aims to provide a preparation method of the 2-methoxy estramustine and the derivatives thereof,

the synthetic routes of the compounds represented by formula 1 and formula 2 are as follows:

the preparation method of the compound shown in the formula 1 comprises the following steps:

dissolving the compound shown in the formula 5 in dichloromethane, adding triethylamine and 4-dimethylaminopyridine, and uniformly stirring; dissolving N, N-bis (2-chloroethyl) carbamoyl chloride in dichloromethane, dropwise adding the dichloromethane into the reaction solution, stirring for reaction for 0.4-0.6 hour, then moving the reaction solution to a water bath with the temperature of 38-42 ℃, keeping the temperature for reaction till the reaction is complete, stopping the reaction, cooling the reaction solution to room temperature, adding distilled water, adjusting the pH of the reaction solution to be 5-6 by using a hydrochloric acid solution under stirring, extracting to obtain an organic phase, and removing the solvent by reduced pressure distillation to obtain the compound shown in the formula 1; wherein the molar ratio of the compound shown in the formula 5 to the triethylamine to the 4-dimethylaminopyridine to the N, N-bis (2-chloroethyl) carbamoyl chloride is as follows: 1 (1.8-2.2), (0.08-0.12), (1.2-1.8);

the preparation method of the compound shown in the formula 2 comprises the following steps:

(1) dropping phosphorus oxychloride into pyridine at 4-6 ℃, and uniformly stirring to obtain a pyridine solution of phosphorus oxychloride; dissolving the compound shown in the formula 1 in pyridine, dropwise adding the pyridine solution into a pyridine solution of phosphorus oxychloride, after the reaction is completed, pouring the reaction solution into an ice-water mixture, adding a hydrochloric acid solution to adjust the pH value to be 2-3, filtering, taking filter residue, drying, dissolving the filter residue in ethyl acetate, adding ethanol, dropwise adding distilled water until the generated turbidity does not disappear, adding a small amount of ethyl acetate to redissolve, continuously stirring, filtering, taking the filter residue, and drying to obtain the compound shown in the formula 6; wherein the mol ratio of the compound shown in the formula 1 to the phosphorus oxychloride is as follows: 1: 4-6

(2) Adding anhydrous methanol into the compound shown in the formula 6, dissolving, and cooling to 0-5 ℃ to obtain a methanol solution of the compound shown in the formula 6; dissolving metal sodium in absolute methanol, and cooling to 5-8 ℃ to obtain a methanol solution of sodium methoxide; dropwise adding a methanol solution of sodium methoxide into a methanol solution of a compound shown in a formula 6, stirring and reacting for 8-15 minutes at 0-5 ℃, adding anhydrous ether at 0-5 ℃, filtering, taking filter residues, washing the filter residues with the anhydrous ether, and drying to obtain a compound shown in a formula 2; wherein, the mol ratio of the compound shown in the formula 6 to the metallic sodium is as follows: 1: 1.1-1.3;

the synthetic route of the compound represented by formula 3 is as follows:

the preparation method of the compound shown in the formula 3 comprises the following steps:

dissolving the compound shown in the formula 1 in dichloromethane, adding triethylamine, stirring uniformly, slowly adding glycyl chloride hydrochloride, stirring until the reaction is complete, adding distilled water, taking an organic phase, washing with inclusion sodium chloride, drying with anhydrous sodium sulfate, distilling under reduced pressure to remove part of a solvent, dropwise adding a HY 1, 4-dioxane solution, stirring, filtering, and taking filter residues to obtain the compound shown in the formula 3; wherein the compound shown in the formula 1, triethylamine, glycinyl chloride hydrochloride and HY have the following molar ratio: 1, (2.5-3.5): (1.2-1.8): 3-5;

the synthetic route of the compound represented by formula 4 is as follows:

the preparation method of the compound shown in the formula 4 comprises the following steps:

dissolving the compound shown in the formula 1 in dichloromethane, adding triethylamine and 4-dimethylaminopyridine, and uniformly stirring; dissolving succinic anhydride or glutaric anhydride in dichloromethane, dropwise adding the succinic anhydride or glutaric anhydride into the reaction solution, stirring until the reaction is complete, stopping the reaction, cooling the reaction solution to room temperature, adding distilled water, adjusting the pH of the reaction solution to be 5-6 by using a hydrochloric acid solution under stirring, extracting to obtain an organic phase, washing, removing the solvent to obtain a solid, dissolving the solid in methanol and water, adding sodium bicarbonate, stirring, distilling under reduced pressure to remove the solvent to obtain a solid, and recrystallizing ethanol and diethyl ether to obtain a compound shown in a formula 4; wherein, the mole ratio of the compound shown in the formula 1, triethylamine, 4-dimethylamino pyridine and succinic anhydride or glutaric anhydride is as follows: 1: (1.8-2.2): (0.08-0.12): (1-1.2).

The invention also aims to provide application of the 2-methoxy estramustine and the derivatives thereof in preparing antitumor drugs.

Preferably, the tumor is prostate cancer, breast cancer or colon cancer.

The invention also aims to provide application of the 2-methoxy estramustine and the derivatives thereof in preparing medicines for treating multiple myeloma.

The invention has the beneficial effects that: the invention provides a novel compound 2-methoxy estramustine and derivatives thereof. The inventor discovers through research that the 2-methoxyestramustine (the compound shown in the formula 1) takes 2-methoxyestradiol as a carrier, an alkylating agent (nitrogen mustard) is connected through carbamate to form a dual-functional drug molecular compound, and the whole molecule is taken as a mitosis inhibitor, so that the 2-methoxyestramustine compound can play a role in tumor cytotoxicity caused by the nitrogen mustard and can play an anti-tumor role of 2-methoxyestradiol released by metabolite mediation. Since 2-methoxyestradiol is an endogenous substance, it has fewer adverse side effects than estradiol, a metabolite of estramustine. The derivative of 2-methoxyestramustine (the compound shown in the formula 2-4) can be used as a prodrug of 2-methoxyestradiol, and aims to improve the water solubility of the compound and further improve the bioavailability. The prodrug is metabolized and hydrolyzed by various active esterases or phosphatases in vivo to release active 2-methoxy estramustine (the compound shown in the formula 1), enters the blood circulation system of a human body and reaches target cells, thereby playing the same anti-tumor role as 2-methoxy estramustine. The invention also provides a preparation method of the 2-methoxy estramustine and the derivatives thereof, and the preparation method can prepare the 2-methoxy estramustine and the derivatives thereof with higher yield. The invention also provides application of the 2-methoxy estramustine and the derivatives thereof in medicaments for treating tumors or multiple myeloma.

Detailed Description

To better illustrate the objects, aspects and advantages of the present invention, the present invention will be further described with reference to specific examples.