Preparation method of vincristine related substance
By reacting vincristine with a specific reaction solvent and catalyst under specific conditions, the vincristine impurities A, impurities D and impurities E recorded in the European Pharmacopoeia were successfully prepared, which solved the complex preparation process in the prior art, achieved an efficient and simple preparation process, and provided reliable impurity standards.
Patent Information
- Application Number
- CN202411905703.9
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-23
- Publication Date
- 2025-03-21
AI Technical Summary
The prior art is difficult to effectively prepare vincristine impurities A, impurities D and impurities E recorded in the European Pharmacopoeia, resulting in a complex preparation process and difficult to obtain impurity standards.
Impurity A, impurity D and impurity E were successfully prepared by adjusting pH, extraction and purification steps. The specific steps include reacting with an oxidation reagent in an aqueous organic acid solution to prepare impurity A, performing a transesterification reaction in an alcohol solvent to prepare impurity D, and performing a transesterification reaction in an alcohol solution to prepare impurity E.
The efficient preparation of vincristine impurities A, impurities D and impurities E is achieved. The process is simple, easy to operate, and the reaction yield is high. The obtained product can be used as impurity standard products to detect and judge vincristine raw materials and its related substances or impurities.
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Figure CN119661561A_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of drug analysis, and in particular relates to a method for preparing vincristine-related substances. Background Art
[0002] Vincristine (Vincristine, molecular formula is C 46 H 56 N4O 10 ), CAS No. 57-22-7, is an indole alkaloid compound extracted from Catharanthus roseus, and its chemical structure is as follows:
[0003] As a microtubule inhibitor, vincristine can bind to tubulin, prevent it from polymerizing into microtubules, interfere with the cell mitosis process, and thus effectively inhibit the proliferation of tumor cells.
[0004] Acute lymphoblastic leukemia (ALL) is the most common hematological malignancy in children and adolescents, characterized by the proliferation of a large number of abnormal lymphoblasts in the bone marrow and peripheral blood. Vincristine plays an irreplaceable role in the treatment of acute lymphoblastic leukemia and is also widely used in chemotherapy regimens for Hodgkin's lymphoma, non-Hodgkin's lymphoma, neuroblastoma and other solid tumors. Compared with traditional chemotherapy drugs, vincristine has higher selectivity, but due to its significant neurotoxicity, the dose needs to be strictly controlled.
[0005] In recent years, research on vincristine has focused on improving drug delivery systems (such as nanoparticle drug delivery systems) and designing new derivatives with lower toxicity and higher efficacy. As a classic anticancer drug, vincristine is not only a core component of existing treatment options, but also plays an important role in promoting the development of new anticancer drugs, and has continued research value and market demand.
[0006] The European Pharmacopoeia (EP) records 8 impurities in the synthesis process of vincristine, including intermediate impurities, isomer impurities, degradation impurities, etc. The structural formulas of impurity A, impurity D, and impurity E are as follows: ; ; . Summary of the invention
[0007] In view of the above-mentioned shortcomings of the prior art, the object of the present invention is to provide a method for preparing impurity A, impurity D and impurity E of vincristine EP (recorded in the European Pharmacopoeia).
[0008] In order to achieve the above-mentioned object and other related objects, the present invention provides the following technical solutions: In the first aspect, the preparation route of impurity A is provided as follows: .
[0009] More specifically, the preparation steps are as follows: Vincristine is dissolved in a reaction solvent and reacted with an oxidizing agent, and then the impurity A is obtained by post-treatment through pH adjustment, extraction, concentration, and purification.
[0010] Furthermore, the reaction solvent comprises an aqueous solution of an organic acid, and the organic acid is selected from one or more of acetic acid, trifluoroacetic acid, trichloroacetic acid, benzoic acid, and o-nitrobenzoic acid, preferably acetic acid or trifluoroacetic acid.
[0011] Furthermore, the oxidizing agent is selected from one or more of hydrogen peroxide, potassium persulfate, benzoyl peroxide, and tert-butyl hydroperoxide, preferably hydrogen peroxide.
[0012] Furthermore, the concentration of the aqueous solution of the organic acid is 0.05% to 5%, preferably 0.05% to 0.5%.
[0013] Furthermore, the reaction temperature is 0-50°C, preferably 20-30°C.
[0014] Furthermore, the reaction time is 24 to 96 hours, preferably 45 to 72 hours.
[0015] It should be noted that, by adopting the above technical scheme, impurity A is obtained by oxidation reaction of vincristine in a low-concentration organic acid aqueous solution.
[0016] In the second aspect, the preparation route of impurity D is provided as follows: .
[0017] More specifically, the preparation steps are as follows: Vincristine is dissolved in a reaction solvent and reacted under the catalysis of a base. After post-treatment, impurity D is obtained through concentration and purification.
[0018] Furthermore, the reaction solvent includes an alcohol solvent, and the alcohol solvent is selected from one or more of methanol, ethanol, n-propanol, and n-butanol, preferably methanol. Methanol includes anhydrous methanol.
[0019] Furthermore, the base-catalyzed reagent includes a non-nucleophilic organic base, and the non-nucleophilic organic base is selected from one or more of triethylamine, DBU (1,8-diazabicyclo[5.4.0]undec-7-ene), diisopropylethylamine, sodium tert-butoxide, and potassium tert-butoxide, preferably DBU.
[0020] Furthermore, the reaction temperature is 0-50° C., and the reaction time is 24-96 hours.
[0021] It should be noted that, by adopting the above technical scheme, impurity D is obtained by subjecting vincristine to an ester exchange reaction in an alcohol solution.
[0022] In a third aspect, the preparation route of impurity E is provided as follows: .
[0023] More specifically, the preparation steps are as follows: Vinblastine is dissolved in a reaction solvent and reacted under the catalysis of a base. After post-treatment, impurity E is obtained through concentration and purification.
[0024] Furthermore, the reaction solvent includes an alcohol solvent, and the alcohol solvent is selected from one or more of methanol, ethanol, n-propanol, and n-butanol, preferably methanol. Methanol includes anhydrous methanol.
[0025] Furthermore, the base-catalyzed reagent includes a non-nucleophilic organic base, and the non-nucleophilic organic base is selected from one or more of triethylamine, DBU (1,8-diazabicyclo[5.4.0]undec-7-ene), diisopropylethylamine, sodium tert-butoxide, and potassium tert-butoxide, preferably DBU.
[0026] Furthermore, the reaction temperature is 0-50° C., and the reaction time is 24-150 hours.
[0027] It should be noted that, by adopting the above technical scheme, impurity E is obtained by subjecting vinblastine to an ester exchange reaction in an alcohol solution.
[0028] The beneficial effects of the present invention are: The present invention adopts corresponding raw materials to prepare impurity A, impurity D and impurity E, has a simple synthesis process, is easy to operate, and has a high reaction yield, can be used as an impurity standard, and provides a detection and judgment basis for subsequent research or monitoring of vincristine raw materials and related substances or impurities. BRIEF DESCRIPTION OF THE DRAWINGS
[0029] The accompanying drawings are used to provide further understanding of the present invention and constitute a part of the specification. They are used to explain the present invention together with the embodiments of the present invention and do not constitute a limitation of the present invention.
[0030] In the attached picture: Figure 1 It is the NMR analysis diagram of impurity A in the present invention; Figure 2 It is the nuclear magnetic analysis diagram of impurity D in the present invention; Figure 3 It is the NMR analysis diagram of impurity E in the present invention. DETAILED DESCRIPTION
[0031] The applicant will describe the embodiments of the present invention in detail below, it should be noted that the embodiments described below are exemplary, are only used to explain the present invention, and cannot be construed as limiting the present invention. In addition, if not otherwise specified, all reagents adopted in the following embodiments are commercially available or can be synthesized with reference to existing literature or known methods, and for the reaction or test conditions not listed, are also the conventional techniques that are easily available to those skilled in the art. The terminology used in the present invention is general in the art, and those skilled in the art can clearly understand its implication, such as, aqueous solution, molten clear, extraction, vacuum distillation, etc., and will not be repeated one by one.
[0032] The present invention provides a method for preparing vincristine-related substances, wherein the vincristine-related substances are EP impurity A, EP impurity D and EP impurity E respectively.
[0033] Example 1 Preparation method of vincristine impurity A: Dissolve 500 mg of vincristine in 50 mL of 0.2% acetic acid aqueous solution, add 15 mL of 5% hydrogen peroxide, and stir at 20-30 °C for 72 hours. Adjust pH to 8.0 with concentrated ammonia, and add 50 mL of DCM (dichloromethane) for extraction. Separate the liquids, extract the aqueous phase with 10 mL of DCM again, and combine the organic phases for vacuum distillation. The evaporated residue is separated and purified by preparative plate to obtain 264 mg of the target product as a light yellow solid, with HPLC purity of 97.69%, MS (ESI): 840.91 [M+H] + . Reference Figure 1 .
[0034] Example 2 Preparation method of vincristine impurity D: Dissolve 200 mg of vincristine in 10 mL of anhydrous methanol, stir until dissolved, add 0.1 mL of DBU, stir at 20-30 °C for 24 hours, and distill the reaction solution under reduced pressure at 45 °C. Evaporate the residue and purify it by preparative plate to obtain 143 mg of the target compound as an off-white solid with HPLC purity of 92.57%, MS (ESI): 782.91 [M+H] + . Reference Figure 2 .
[0035] Example 3 Preparation method of vincristine impurity E: Dissolve 600 mg of Vinblastine in 30 mL of anhydrous methanol, stir until dissolved, add 0.5 mL of DBU, stir at 20-30 °C for 6 days, and distill the reaction solution under reduced pressure at 45 °C. Evaporate the residue and purify it by preparative liquid phase separation to obtain 249 mg of the target compound as an off-white solid with HPLC purity of 95.82%, MS (ESI): 768.91 [M+H] + . Reference Figure 3 .
[0036] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention is described in detail with reference to the aforementioned embodiments, those skilled in the art can still modify the technical solutions described in the aforementioned embodiments or replace some of the technical features therein by equivalents. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.
Claims
1. A method for preparing vincristine-related substances, wherein the vincristine-related substances include EP impurities of vincristine, and the EP impurities include impurity A, impurity D, and impurity E, characterized in that: The preparation route of impurity A is as follows: ； The preparation route of impurity D is as follows: ； The preparation route of impurity E is as follows: 。 2. The method for preparing vincristine-related substances according to claim 1, characterized in that: The preparation of impurity A includes the following preparation steps: Vincristine is dissolved in a reaction solvent and reacts with an oxidizing agent; after the reaction, impurity A is obtained by post-treatment through pH adjustment, extraction, concentration, and purification.
3. The method for preparing vincristine-related substances according to claim 2, characterized in that: In the preparation of impurity A, the reaction solvent includes an aqueous solution of an organic acid, and the organic acid includes one or more of acetic acid, trifluoroacetic acid, trichloroacetic acid, benzoic acid, and o-nitrobenzoic acid; the concentration of the aqueous solution of the organic acid is 0.05% to 5%.
4. The method for preparing vincristine-related substances according to claim 2, characterized in that: In the preparation of impurity A, the oxidizing agent includes one or more of hydrogen peroxide, potassium persulfate, benzoyl peroxide, and tert-butyl hydroperoxide; the reaction temperature is 0~50°C, and the reaction time is 24~96 hours.
5. The method for preparing vincristine-related substances according to claim 1, characterized in that: The preparation of impurity D includes the following preparation steps: Vincristine is dissolved in a reaction solvent and reacts under the catalysis of a base. After the reaction, impurity D is obtained through post-treatment, concentration and purification.
6. The method for preparing vincristine-related substances according to claim 5, characterized in that: In the preparation of impurity D, the reaction solvent includes an alcohol solvent, and the alcohol solvent includes one or more of methanol, ethanol, n-propanol, and n-butanol.
7. The method for preparing vincristine-related substances according to claim 5, characterized in that: In the preparation of impurity D, the base-catalyzed reagent includes a non-nucleophilic organic base, and the non-nucleophilic organic base includes one or more of triethylamine, DBU, diisopropylethylamine, sodium tert-butoxide, and potassium tert-butoxide.
8. The method for preparing vincristine-related substances according to claim 1, characterized in that: The preparation of impurity E includes the following preparation steps: Vinblastine is dissolved in a reaction solvent and reacts under the catalysis of a base. After the reaction, impurity E is obtained through post-treatment, concentration and purification.
9. The method for preparing vincristine-related substances according to claim 8, characterized in that: In the preparation of impurity E, the reaction solvent includes an alcohol solvent, and the alcohol solvent includes one or more of methanol, ethanol, n-propanol, and n-butanol.
10. The method for preparing vincristine-related substances according to claim 8, characterized in that: In the preparation of impurity E, the base-catalyzed reagent includes a non-nucleophilic organic base, and the non-nucleophilic organic base includes one or more of triethylamine, DBU, diisopropylethylamine, sodium tert-butoxide, and potassium tert-butoxide.