CN102266568A - Preparation method for hydroxypropyl cyclodextrin inclusion of taxol - Google Patents

Abstract

The invention discloses a preparation method for a hydroxypropyl cyclodextrin inclusion of taxol, and belongs to the technical field of preparation of water-soluble medicaments in the technical field of super-molecular inclusion. Hydroxypropyl-cyclodextrin serving as host molecules, taxol serving as guest molecules and N,N-dimethyl formamide serving as a reaction solvent are subjected to homogeneous super-molecular reaction to generate the water-soluble taxol super-molecular inclusion. The purity of the inclusion product prepared by the method is high; and the method is simple and convenient in operation, controllable in conditions and easy for industrialized production. The prepared taxol-cyclodextrin super-molecular inclusion has good water solubility, the solubility of the inclusion in the water of 25 DEG C is 0.26mg/ml, the structure of the inclusion is stable, and the structure and the medicinal performance of the taxol are not destroyed. Due to excellent water solubility of the taxol-hydroxypropyl cyclodextrin inclusion, the taxol which is a natural anti-cancer medicament with poor water solubility is expected to become an oral medicament or intravenous injection.

Description

A kind of preparation method of hydroxypropyl cyclodextrin inclusion compound of paclitaxel

technical field

The invention belongs to the technical field of preparation of water-soluble drugs in the technical field of supramolecular inclusion.

Background technique

Paclitaxel is a complex diterpenoid compound, molecular formula: C ₄₇ H ₅₁ NO ₁₄ , molecular weight: 853.918, white crystal. Paclitaxel is relatively stable in the pH range of 4-8, decomposes quickly under alkaline conditions, and relatively stable under acidic conditions. Paclitaxel is a natural anti-tumor substance isolated from the bark of the yew tree. The unique anticancer activity mechanism of paclitaxel is that it belongs to mitosis inhibitor or spindle toxin, which can not only inhibit cell mitosis, the formation of spindle and spindle filament, thereby preventing the proliferation of cancer cells, but also induce and promote the formation of tubulin. Polymerization and assembly prevent microtubules from depolymerizing and stabilize microtubules, so they are active against many tumors resistant to conventional chemotherapy drugs, and have broad application prospects. However, paclitaxel is difficult to dissolve in water and many pharmaceutical solvents at room temperature, the solubility in water is only 0.006 mg·mL ^-1 , it is difficult to absorb orally, and it is usually administered by intravenous injection. Currently, paclitaxel preparations used clinically are mainly prepared by mixing polyoxyethylene castor oil (CrEL) and absolute ethanol (50:50, V/V) as solvents to make injections. Due to the large amount of CrEL in this preparation, severe allergic reactions will occur after intravenous infusion, so before administration, patients need to pre-administer corticosteroids and antihistamines to reduce allergic reactions. Paclitaxel formulations with CrEL-absolute ethanol as a solvent must be diluted to 0.3-1.2 mg/ml for infusion, but the diluent can only remain stable within 12-24 hours.

As the second-generation supramolecular main compound—cyclodextrins (CDs for short), a class of cyclic oligosaccharides composed of more than 6 D-glucopyranose units bonded by а-1,4, according to Depending on the number of glucose units it contains, it can be divided into а-CD, β-CD and γ-CD, etc. Among them, β-cyclodextrin (β-CD) has the highest yield, the cheapest price, and the most widely used. The characteristic of cyclodextrin compounds is that they all have a certain size of hydrophobic cavity, so they can selectively include various types of guest molecules to form supramolecular compounds. Due to its unique structure, both theoretical research and practical application of cyclodextrin have been developed rapidly. It has important uses in industry, agriculture, food, medicine, separation technology and environmental protection.

-环糊精（HP--CD）被认为是最有用的环糊精衍生物之一，是美国食品和药物管理局(Food and Drug Аdministration，FDА)批准的第一个可供静脉注射的

-CD衍生物，具有毒性低、溶血性低和水溶性好的特点。HP-

-CD是

-CD与1,2-环氧丙烷缩合而成的亲水性衍生物。通过对

-CD的羟丙基化，破坏其分子内氢键，使其水溶性显著提高。

-CD的水溶性在室温下约为1.85％(w/v)，而HP-

-CD则易溶于水，室温下溶解度>50％(w/v)，甚至可以高达75％(w/v)以上，当其浓度<40％(w/v)，流动性好，不粘稠。高的水溶性和分子柔性，使得HP--CD能很好地与药物分子包合形成非共价复合物，因此可以提高药物的稳定性、水溶性，降低药物的挥发性，控制药物释放速率，掩盖不良气味等。另外这种包合材料具有肾毒性低、溶血作用小、局部刺激性轻微等特点, 这使难溶性药物制成注射剂成为可能。 The unique cage structure of β-cyclodextrin can include drug molecules to form an inclusion compound. At this time, the drug molecules are contained in the molecular cavity of β-CD, which has a high degree of dispersion. The hydrophilicity makes the inclusion compound have good wettability, so as to achieve the solubilization effect on poorly soluble drugs. Due to the low solubility of β-CD itself, modifying groups are often introduced at its edge to improve its solubility and expand its application range. Among many cyclodextrin derivatives, hydroxypropyl-

-Cyclodextrin (HP- -CD) is considered to be one of the most useful cyclodextrin derivatives and is the first approved by the U.S. Food and Drug Аadministration (FDА) for intravenous administration

-CD derivative, with low toxicity, low hemolysis and good water solubility. HP-

- CD is

-A hydrophilic derivative formed by condensation of CD and 1,2-propylene oxide. by right

-The hydroxypropylation of CD destroys its intramolecular hydrogen bonds and significantly improves its water solubility.

The water solubility of -CD is about 1.85% ( w/v ) at room temperature, while HP-

-CD is easily soluble in water, the solubility at room temperature is >50% ( w/v ), and can even be as high as 75% ( w/v ), when its concentration is <40% ( w/v ), it has good fluidity and is not sticky thick. High water solubility and molecular flexibility make HP- -CD can be well included with drug molecules to form non-covalent complexes, so it can improve the stability and water solubility of drugs, reduce the volatility of drugs, control the release rate of drugs, and mask bad odors. In addition, this inclusion material has the characteristics of low nephrotoxicity, small hemolysis, and mild local irritation, which makes it possible for poorly soluble drugs to be made into injections.

Currently, paclitaxel-hydroxypropyl- -The preparation method of cyclodextrin inclusion compound has been reported, mainly adopting mixed solvent method to carry out inclusion reaction. Its preparation method: the mass ratio of paclitaxel to cyclodextrin is 1:10~150, the ethanol solution of paclitaxel is added dropwise in the cyclodextrin aqueous solution, the system is dissolved and then filtered with a 0.2~0.4 μm microporous membrane, and the filtrate is decompressed to remove Ethanol, dehydration under reduced pressure, freeze-drying to obtain solid clathrate, or depressurize to obtain liquid clathrate, wherein the ethanol content is less than 2%.

The defective of prior art is:

1. During the preparation process, in order to make the drug be wrapped in the cyclodextrin cavity as much as possible, the amount of the main compound is often much greater than the amount of the drug (mass ratio 10~150:1), resulting in the final product containing a large amount of The main compound cannot guarantee the purity of the clathrate, which affects the determination of the drug concentration in the next step, and is also a waste of raw materials.

2. The supramolecular clathrate is prepared by the mixed solvent method. Since the solubility of paclitaxel in water is extremely small, some paclitaxel cannot be dissolved in the mixed solvent, and the inclusion reaction is not carried out in a homogeneous system. On the one hand, the inclusion reaction It cannot be fully carried out, and the yield is low; on the other hand, after the inclusion reaction is completed, the reaction system needs to be filtered with a microporous membrane to remove unreacted or dissolved paclitaxel, which virtually increases the cost of inclusion compound preparation.

3. The mixed solvent method is used, the operation is relatively complicated, the follow-up treatment is relatively cumbersome, the energy consumption is high, and ethanol and water need to be distilled under reduced pressure.

Contents of the invention

The purpose of the present invention is to propose a method for preparing a novel water-soluble paclitaxel-cyclodextrin supramolecular inclusion compound, aiming at overcoming the above defects and adopting a simpler method to prepare a high-purity inclusion compound.

-环糊精和紫杉醇溶解于N，N-二甲基甲酰胺中，在温度为15～35 ℃的条件下，搅拌反应；待反应结束后，再加入甲醇，析出白色固体；将白色固体抽滤，滤饼经无水乙醇洗涤后置于30～50℃真空干燥，得到水溶性紫杉醇-羟丙基-

-环糊精包合物。 The technical scheme of the present invention is: the hydroxypropyl-

- Dissolve cyclodextrin and paclitaxel in N,N-dimethylformamide, and react with stirring at a temperature of 15-35°C; after the reaction is complete, add methanol to precipitate a white solid; pump the white solid After filtering, the filter cake was washed with absolute ethanol and then vacuum-dried at 30-50°C to obtain water-soluble paclitaxel-hydroxypropyl-

- Cyclodextrin inclusion complexes.

-环糊精为主体分子、紫杉醇为客体分子、N，N-二甲基甲酰胺为反应溶剂，发生均相超分子反应生成水溶性紫杉醇超分子包合物。DMF可以同时溶解羟丙基--环糊精与紫杉醇，使得包合反应在均相溶液中进行，提高反应的效率。该方法制备包合物产品纯度高，操作简便，条件易控，易于工业化生产。使用羟丙基-

-环糊精与紫杉醇的共溶溶剂，使得包合反应在均相溶液中。利用包合物在甲醇中溶解度较差，以甲醇为沉淀剂，使包合物沉淀，再经过抽滤、洗涤、干燥后，即可获得紫杉醇-羟丙基环糊精包合物。制备的紫杉醇-环糊精超分子包合物水溶性好，于25℃水中的溶解度为0.26 mg/ml，结构稳定，不破坏紫杉醇自身的结构和药用性能。紫杉醇-羟丙基环糊精包合物优良的水溶性，使得紫杉醇这种水溶性较差的天然抗癌药物，有望做成口服药物或静脉注射针剂。 The present invention uses hydroxypropyl-

-Cyclodextrin is the host molecule, paclitaxel is the guest molecule, and N,N-dimethylformamide is the reaction solvent, and a homogeneous supramolecular reaction occurs to generate the water-soluble paclitaxel supramolecular inclusion compound. DMF can simultaneously dissolve hydroxypropyl- - Cyclodextrin and paclitaxel enable the inclusion reaction to proceed in a homogeneous solution, improving the efficiency of the reaction. The clathrate prepared by the method has high purity, simple operation, easy control of conditions, and easy industrial production. Use hydroxypropyl-

- A co-solvent for cyclodextrin and paclitaxel, making the inclusion reaction in a homogeneous solution. Taking advantage of the poor solubility of the clathrate in methanol, methanol is used as a precipitant to precipitate the clathrate, and after suction filtration, washing and drying, the paclitaxel-hydroxypropyl cyclodextrin clathrate can be obtained. The prepared paclitaxel-cyclodextrin supramolecular inclusion compound has good water solubility, the solubility in water at 25°C is 0.26 mg/ml, the structure is stable, and the structure and medicinal properties of paclitaxel are not destroyed. The excellent water solubility of paclitaxel-hydroxypropyl cyclodextrin inclusion compound makes paclitaxel, a natural anticancer drug with poor water solubility, hopeful to be made into oral medicine or intravenous injection.

-环糊精和紫杉醇的投料摩尔比为1︰5～10。 In order to improve the purity of clathrate, the hydroxypropyl-

- The molar ratio of cyclodextrin and paclitaxel is 1:5-10.

Description of drawings

Fig. 1 is the infrared spectrogram of the hydroxypropyl cyclodextrin inclusion complex of water-soluble paclitaxel and the host-guest compound prepared by the method of the present invention.

Detailed ways

In order to make the object, technical solution and advantages of the present invention clearer, the present invention will be described in detail below in conjunction with examples.

-环糊精和0.5～1 mol紫杉醇，主客体分子摩尔比为1︰5～10，溶解于100 ml N，N-二甲基甲酰胺(DMF)。反应温度15～35 ℃，磁力搅拌或机械搅拌24 h。 1. Add 0.1 mol hydroxypropyl-

- Cyclodextrin and 0.5-1 mol paclitaxel, the host-guest molecular molar ratio is 1:5-10, dissolved in 100 ml N,N-dimethylformamide (DMF). The reaction temperature is 15-35 ℃, magnetic stirring or mechanical stirring for 24 h.

-环糊精包合物。 2. After the reaction is over, add 20-50 ml of precipitant methanol into the Erlenmeyer flask, and a white solid will precipitate out of the reaction system. Suction filter the solid, retain the filtrate and transfer it to a round bottom flask, wash the filter cake with a small amount of absolute ethanol, and then place the solid at 30-50 °C for 24 h in vacuum to obtain a white solid, which is water-soluble paclitaxel-hydroxypropyl -

- Cyclodextrin inclusion complexes.

3. The above filtrate transferred to the round bottom flask is first subjected to vacuum distillation to remove the methanol in the filtrate (the distilled methanol can also be recycled), and the remaining filtrate contains a large amount of unincluded guest molecule paclitaxel. Can continue to add hydroxypropyl- - Cyclodextrin, carry out inclusion reaction, repeat the above operation without changing the solvent. The amount concentration of the substance of the guest molecule should be kept more than twice that of the substance of the host molecule. There is almost no solvent loss in the whole process.

-环糊精的空腔中，形成了超分子包合物。 4. The product is identified by infrared spectroscopy. Infrared spectra were measured by Bruker-Tensor 27 infrared spectrometer. As shown in Figure 1, (a) in the figure is the infrared spectrum of paclitaxel. The carbonyl-C=O vibration and double bond-C=C vibration absorption peaks of paclitaxel are at 1731 cm ^-1 and 1648 cm ^-1 respectively. (b) is the infrared spectrum of hydroxypropyl cyclodextrin. (c) is the infrared spectrogram of paclitaxel-hydroxypropyl cyclodextrin inclusion complex, and the carbonyl and C=C double bond vibration absorption peaks appear at the same wave number in (c), thus indicating that paclitaxel has entered into hydroxypropyl cyclodextrin base-

- In the cavity of the cyclodextrin, a supramolecular clathrate is formed.

Claims (2)

1. a preparation method of a hydroxypropyl cyclodextrin inclusion compound of paclitaxel, characterized in that: hydroxypropyl-

- Dissolve cyclodextrin and paclitaxel in N,N-dimethylformamide, and react with stirring at a temperature of 15-35°C; after the reaction is complete, add methanol to precipitate a white solid; pump the white solid After filtering, the filter cake was washed with absolute ethanol and then vacuum-dried at 30-50°C to obtain water-soluble paclitaxel-hydroxypropyl-

- Cyclodextrin inclusion complexes. 2. according to the preparation method of the hydroxypropyl cyclodextrin clathrate of paclitaxel described in claim 1, it is characterized in that: described hydroxypropyl-

- The molar ratio of cyclodextrin and paclitaxel is 1:5-10.

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