阅读说明：本技术 一种10-羰基多西他赛的制备方法 (Preparation method of 10-carbonyl docetaxel ) 是由 黄春 陆叶梦 王莉佳 王旭阳 于 2021-09-02 设计创作，主要内容包括：本发明公开了一种10-羰基多西他赛的制备方法,其技术方案包括以下步骤：S1：将多西他赛用溶剂溶清,加入重金属盐,搅拌反应,反应完全后,抽滤,浓缩有机相得降解混合物；S2：降解混合物进行柱层析纯化,流动相为二氯甲烷、甲醇和乙酸,收集产物段浓缩,用环己烷打浆,抽滤干燥即得10-羰基多西他赛,本发明的优点在于通过高效降解和巧妙的纯化设计,有较高的收率,操作简易,产品纯度好。(The invention discloses a preparation method of 10-carbonyl docetaxel, which comprises the following steps: s1: dissolving docetaxel in a solvent, adding heavy metal salt, stirring for reaction, performing suction filtration after the reaction is completed, and concentrating an organic phase to obtain a degradation mixture; s2: the degraded mixture is purified by column chromatography, the mobile phase is dichloromethane, methanol and acetic acid, the product is collected and concentrated, the product is pulped by cyclohexane, and the 10-carbonyl docetaxel is obtained by suction filtration and drying.)

1. A preparation method of 10-carbonyl docetaxel is characterized by comprising the following steps:

s1: dissolving docetaxel in a solvent, adding heavy metal salt, stirring for reaction, performing suction filtration after the reaction is completed, and concentrating an organic phase to obtain a degradation mixture;

s2: purifying the degraded mixture by column chromatography, collecting the product, concentrating, pulping with cyclohexane, and vacuum filtering to obtain 10-carbonyl docetaxel;

the reaction route of the preparation process is as follows:

2. the method for preparing 10-carbonyl docetaxel according to claim 1, wherein: in step S1, the solvent is one or more of methanol, ethanol, and N, N-dimethylformamide.

3. The method for preparing 10-carbonyl docetaxel according to claim 1, wherein: in step S1, the heavy metal salt is one of copper acetate, copper carbonate, manganese (III) acetate, cobalt (III) acetate, and silver carbonate, and the weight ratio of docetaxel to the heavy metal salt is 3-8.

4. The method for preparing 10-carbonyl docetaxel according to claim 1, wherein: in step S1, the reaction temperature is 65-80 ℃ and the reaction time is 3-8 h.

5. The method for preparing 10-carbonyl docetaxel according to claim 1, wherein: in step S2, the dichloromethane is frozen in advance, and the temperature condition of the frozen dichloromethane is below 0 ℃.

6. The method for preparing 10-carbonyl docetaxel according to claim 1, wherein: in step S2, the volume ratio of dichloromethane, methanol and acetic acid is 100:1: 0.1.

7. The method for preparing 10-carbonyl docetaxel according to claim 3, wherein: in step S1, docetaxel is dissolved in methanol, and then a heavy metal salt, which is copper acetate, is added, and the mixture is heated to 65 ℃ and stirred for reflux reaction for 8 hours.

8. The method for preparing 10-carbonyl docetaxel according to claim 3, wherein: in step S1, docetaxel is dissolved in N, N-dimethylformamide, and then a heavy metal salt, which is copper carbonate, is added, and the mixture is heated to 80 ℃ and stirred to reflux and react for 3 hours.

9. The method for preparing 10-carbonyl docetaxel according to claim 3, wherein: in step S1, docetaxel is dissolved in ethanol, and then a heavy metal salt, which is cobalt (III) acetate, is added, and the mixture is heated to 78 ℃ and stirred for reflux reaction for 8 hours.

10. The method for preparing 10-carbonyl docetaxel according to claim 3, wherein: in step S1, docetaxel is dissolved in N, N-dimethylformamide, and then a heavy metal salt, which is silver carbonate, is added, and the mixture is heated to 80 ℃ and stirred to reflux and react for 3 hours.

Technical Field

The invention relates to the field of anti-cancer drugs, in particular to a preparation method of 10-carbonyl docetaxel.

Background

Docetaxel, a taxane drug, has the pharmacological effect of inhibiting mitosis and proliferation of cancer cells by promoting the assembly of microtubule dimers into miniducts, simultaneously stabilizing the miniducts by preventing the process of depolymerization, arresting the cells in G2 and M phases.

One degradation product of docetaxel is 10-carbonyl docetaxel, which is recorded as impurity B by European Pharmacopoeia (EP). In order to better meet international standards, intensive studies on the impurities are also required in the quality studies of docetaxel, and thus it is very necessary to prepare high-purity 10-carbonyl docetaxel. The preparation of 10-carbonyl docetaxel is mainly achieved by degradation, which is always accompanied by the production of 7-epi-10-carbonyl docetaxel (docetaxel impurity D). The 10-carbonyl docetaxel has special properties, and is very easy to be further degraded into 7-epi-10-carbonyl docetaxel after the purity is high, so that the conventional separation method is difficult to realize the preparation of the high-purity 10-carbonyl docetaxel.

Disclosure of Invention

Aiming at the defects of the prior art, the invention aims to provide a preparation method of 10-carbonyl docetaxel, which has the advantages of high yield, simple operation and good product purity through efficient degradation and ingenious purification design.

The technical purpose of the invention is realized by the following technical scheme:

a preparation method of 10-carbonyl docetaxel comprises the following steps:

s1: dissolving docetaxel in a solvent, adding heavy metal salt, stirring for reaction, performing suction filtration after the reaction is completed, and concentrating an organic phase to obtain a degradation mixture;

s2: purifying the degraded mixture by column chromatography, collecting the product, concentrating, pulping with cyclohexane, and vacuum filtering to obtain 10-carbonyl docetaxel;

the reaction route of the preparation process is as follows:

further, in step S1, the solvent is one or more of methanol, ethanol, and N, N-dimethylformamide.

Further, in step S1, the heavy metal salt is one of copper acetate, copper carbonate, manganese (III) acetate, cobalt (III) acetate, and silver carbonate.

Further, in step S1, the reaction temperature is 65-80 ℃, and the reaction time is 3-8 h.

Further, in step S2, the dichloromethane is frozen in advance, and the temperature condition of the dichloromethane is 0 ℃ or lower.

Further, in step S2, the volume ratio of dichloromethane, methanol and acetic acid is 100:1: 0.1.

Further, in step S1, docetaxel was dissolved in methanol, followed by addition of copper acetate, heating to 65 ℃ and reflux reaction with stirring for 8 hours.

Further, in step S1, docetaxel was dissolved in N, N-dimethylformamide, copper carbonate was added, and the mixture was heated to 80 ℃ and stirred to reflux and react for 3 hours.

Further, dissolving docetaxel in ethanol, adding heavy metal salt, namely cobalt (III) acetate, heating to 78 ℃, and carrying out reflux reaction for 8 hours under stirring.

Further, dissolving docetaxel in N, N-dimethylformamide, adding heavy metal salt, wherein the heavy metal salt is silver carbonate, heating to 80 ℃, and carrying out reflux reaction for 3 hours under stirring.

In conclusion, the invention has the following beneficial effects:

1. through screening several heavy metal salts, 10-carbonyl docetaxel can be obtained by selectively oxidizing 10-hydroxyl, main products in the reaction are only 10-carbonyl docetaxel and 7-epi-10-carbonyl docetaxel, the raw materials are completely degraded, the product types are few, the industrial production is facilitated, and the method has important significance for the quality research of docetaxel.

2. Avoids the simultaneous oxidation of hydroxyl at the 7-position and the 2' -position of the side chain caused by the traditional strong oxidant, and the increase of products and the influence on the final yield.

Drawings

FIG. 1 is an HPLC chromatogram of 10-carbonyl docetaxel obtained in example 1 of the present invention.

FIG. 2 is an MS spectrum of 10-carbonyl docetaxel obtained in example 1 of the present invention.

FIG. 3 is a 1H NMR spectrum of 10-carbonyl docetaxel obtained in example 1 of the present invention.

FIG. 4 is a 13C NMR spectrum of 10-carbonyl docetaxel obtained in example 1 of the present invention.

FIG. 5 is an HPLC chromatogram of 10-carbonyl docetaxel obtained in comparative example 3 of the present invention

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the following detailed description of the present invention is provided with reference to the accompanying drawings and the detailed description. The advantages and features of the present invention will become more apparent from the following description. It should be understood that the drawings and figures are only for the purpose of illustration and description, and are not intended to limit the scope of the invention, which is defined by the claims, but rather by the claims.

Example 1:

step S1: dissolving 10g of docetaxel in 200ml of methanol, adding 2g of copper acetate, heating to 65 ℃, and carrying out reflux reaction for 8 hours under the condition of stirring. After the reaction is completed, the reaction solution is filtered, and the filtrate is concentrated to obtain a degradation mixture.

S2: dissolving the degraded mixture with a small amount of dichloromethane, carrying out wet loading, carrying out column chromatography purification (common 200-300 mesh silica gel), wherein the mobile phase comprises dichloromethane, methanol and acetic acid (the volume ratio is 100:1:0.1), the dichloromethane is pre-frozen (below 0 ℃), collecting the product, concentrating, adding a small amount of cyclohexane, pulping, carrying out suction filtration, and drying to obtain 3.4g of 10-carbonyl docetaxel, the purity is 97.366%, and the liquid phase spectrum is shown in figure 1.

The structure of 10-carbonyl docetaxel is shown as the formula I:

as shown in fig. 2, the MS spectrum of 10-carbonyl docetaxel in this example, wherein the mass/charge ratio M/z is 828.3[ M + Na ] +, the molecular weight is estimated to be 805, consistent with the target compound.

As shown in fig. 3, the 1H NMR spectrum of 10-carbonyl docetaxel of this example is 1H NMR (500MHz, CDCl3) δ:7.31 to 8.12(10H, ArH),6.22(1H, t, J ═ 8.9Hz, H13),5.81(1H, d, J ═ 6.8Hz, H35),5.39(1H, H30),5.26(1H, H2),4.90(1H, d, J ═ 8.6Hz, H5),4.63(1H, s, H29),4.31(1H, d, J ═ 8.5Hz, H20a),4.18(1H, d, J ═ 8.5Hz, H20b),4.02(1H, ddt, J ═ 10.5Hz,7.1H, 7H, H8962, cycot (1H, H586H, H5966H, H3H 2H, solvet, H3H, H3H, H3H, H3, 1.29(3H, s, H16/H17),1.26(3H, s, H16/H17);

as shown in FIG. 4, the 13C NMR spectrum of this example 10-carbonyl docetaxel was obtained by 13C NMR (125MHz, CDCl3) delta: 206.116,193.962,172.753,170.167,166.972,155.468,146.996,141.655,138.233,133.874,130.207,128.937,128.896,128.771,128.148,126.754,84.231,80.661,80.344,79.264,76.153,74.66,73.521,72.105,68.705,58.392,56.315,45.35,40.912,35.578,35.403,28.178,26.898(Solvent Cyclohexane),23.887,22.493,14.186,8.362.

Example 2:

step S1: dissolving 10g of docetaxel in 200ml of N, N-dimethylformamide, adding 2g of copper carbonate, heating to 80 ℃, and carrying out reflux reaction for 3 hours under stirring. After the reaction is completed, the reaction solution is filtered, and the filtrate is concentrated to obtain a degradation mixture.

S2: dissolving the degraded mixture with a small amount of dichloromethane, carrying out wet loading, carrying out column chromatography purification, wherein the mobile phase comprises dichloromethane, methanol and acetic acid (the volume ratio is 100:1:0.1), the dichloromethane is pre-frozen (below 0 ℃), collecting the product, concentrating, adding a small amount of cyclohexane, pulping, carrying out suction filtration, and drying to obtain 2.8g of 10-carbonyl docetaxel, the purity is 96.8%, and the total yield is 28%.

Example 3:

step S1: dissolving 10g of docetaxel in 200ml of ethanol, adding 2g of cobalt (III) acetate, heating to 78 ℃, and carrying out reflux reaction for 8 hours under stirring. After the reaction is completed, the reaction solution is filtered, and the filtrate is concentrated to obtain a degradation mixture.

S2: dissolving the degraded mixture with a small amount of dichloromethane, carrying out wet loading, carrying out column chromatography purification (common 200-300 mesh silica gel), wherein the mobile phase comprises dichloromethane, methanol and acetic acid (the volume ratio is 100:1:0.1), the dichloromethane is pre-frozen (below 0 ℃), collecting the product, concentrating, adding a small amount of cyclohexane, pulping, carrying out suction filtration, and drying to obtain 3.8g of 10-carbonyl docetaxel, the purity is 97.8%, the total yield is 34%, and the total yield is 38%.

Example 4:

step S1: dissolving 10g of docetaxel in 200ml of N, N-dimethylformamide, adding 2g of silver carbonate, heating to 80 ℃, and carrying out reflux reaction for 3 hours under stirring. After the reaction is completed, the reaction solution is filtered, and the filtrate is concentrated to obtain a degradation mixture.

S2: dissolving the degraded mixture with a small amount of dichloromethane, carrying out wet loading, carrying out column chromatography purification, wherein the mobile phase comprises dichloromethane, methanol and acetic acid (the volume ratio is 100:1:0.1), the dichloromethane is pre-frozen (below 0 ℃), collecting the product, concentrating, adding a small amount of cyclohexane, pulping, carrying out suction filtration, and drying to obtain 0.8g of 10-carbonyl docetaxel, and the purity is 95.3%. The yield is low, and the main product of the reaction is 7-epidocetaxel.

Comparative example 1 (using hydrogen peroxide as oxidant):

dissolving 2g of docetaxel in 40ml of methanol, adding 2ml of 30% hydrogen peroxide, heating to 65 ℃, and carrying out reflux reaction for 3 hours under stirring. The product obtained after the raw materials are reacted is mainly a side chain hydrolysis product, and the target 10-carbonyl docetaxel is basically not observed.

Comparative example 2 (using manganese dioxide as the oxidant):

2g of docetaxel is dissolved and cleared by 40ml of methanol, 0.5g of manganese dioxide is added, and the mixture is heated to 65 ℃ and stirred for reflux reaction for 3 hours. The product obtained after the raw material reaction is almost 7, the 10-carbonyl docetaxel accounts for a small amount, and the separation is difficult.

Comparative example 3 (using lead dioxide as the oxidant):

dissolving 2g of docetaxel in 40ml of N, N-dimethylformamide, adding 0.5g of lead dioxide, heating to 65 ℃, and carrying out reflux reaction for 3 hours under stirring. The number of products obtained after the raw materials are reacted is approximately 7, and the 10-carbonyl docetaxel accounts for a small amount.

Comparative example 4 (using cobalt trifluoride oxidizer):

dissolving 2g of docetaxel in 40ml of N, N-dimethylformamide, adding 0.5g of cobalt trifluoride, heating to 65 ℃, and carrying out reflux reaction for 3 hours under stirring. The products obtained after the raw materials are reacted are complex, more than ten products exist, and 10-carbonyl docetaxel is not found.

Comparative example 5 (purification using column chromatography coupled with preparative liquid phase by conventional means):

step S1: dissolving 10g of docetaxel in 200ml of methanol, adding 2g of copper acetate, heating to 65 ℃, and carrying out reflux reaction for 8 hours under the condition of stirring. After the reaction is completed, carrying out suction filtration on the reaction solution, and concentrating the filtrate to obtain a degradation mixture;

s2: and dissolving the degraded mixture by using a small amount of dichloromethane, then carrying out wet loading, carrying out column chromatography purification (common 200-300 mesh silica gel), wherein the mobile phase is dichloromethane and methanol (the volume ratio is 100:1), completely crossing 10-carbonyl docetaxel and 7-epi-10-carbonyl docetaxel after 7-epi-10-carbonyl docetaxel is eluted in the column chromatography process, and collecting and concentrating to obtain the 10-carbonyl docetaxel with lower purity.

Dissolving the 10-carbonyl docetaxel in a small amount of acetonitrile, performing liquid phase purification (the filler is conventional C-18 reversed phase filler), increasing the acetonitrile by 50-70%, collecting eluent of the first main peak (the elution is completed within 30 min), concentrating the eluent to separate out solid, performing suction filtration and drying to obtain 10-carbonyl docetaxel with the purity of 28.261%, and obtaining a map shown in figure 5, wherein most target products are converted into degradation products of 7-epi-10-carbonyl docetaxel.

From the comparative examples it can be seen that: 1. the conventional oxidation reagent can not degrade the docetaxel into 10-carbonyl docetaxel in a large proportion; 2. performing silica gel column chromatography at normal temperature, wherein the 10-carbonyl docetaxel is further degraded in the elution process to obtain 7-epi-10-carbonyl docetaxel; 3. the preparation of liquid phase (C-18 reverse phase packing) liquid could not achieve effective purification of 10-carbonyl docetaxel even though the contact time of 10-carbonyl docetaxel with packing was less than 30min in the whole process.

According to the invention, the degradation of 10-carbonyl docetaxel can be effectively inhibited by a low-temperature mobile phase and the addition of a stabilizer acetic acid, so that the 10-carbonyl docetaxel with high purity is finally obtained.

The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.

The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.