CN100508969C - Nano particles of taxane cyclodextrin inclusion compound and preparation method thereof - Google Patents

Abstract

The invention discloses a taxane, especially a paclitaxel cyclodextrin inclusion complex nanoparticle and a pharmaceutical preparation thereof. Using hydroxypropyl-β-cyclodextrin as a carrier and povidone as a stabilizer, paclitaxel inclusion complex nanoparticles are prepared, with a particle size of 40-200nm. The cyclodextrin inclusion complex nanoparticles increase the solubility of paclitaxel up 100-500 times. Animal experiments showed that compared with the commercially available paclitaxel, the tolerated dose was increased by 2.67 times, and the tumor-inhibiting effect was significantly enhanced. The invention adopts conventional auxiliary materials, has simple process conditions, is suitable for industrial-scale production, and has stable product quality. It can be directly or secondary processed into injection or oral preparations, thereby providing a new and effective anticancer drug taxane for clinical use. way.

Description

Taxane cyclodextrin inclusion complex nanoparticles and preparation method thereof

technical field

The invention relates to taxane cyclodextrin inclusion compound nanoparticles and a preparation method thereof, in particular to the preparation of taxane inclusion compound nanoparticles by hydroxypropyl-β-cyclodextrin. Cyclodextrin inclusion can solubilize and slow-release taxanes, thereby enhancing drug efficacy, thus providing a new and feasible way for anticancer drug taxanes to be used clinically.

technical background

Paclitaxel (Taxol) is a natural product isolated from the bark of the Pacific Yew tree. Paclitaxel has good anticancer activity and has become the third-generation antineoplastic drug after doxorubicin and cisplatin. It is toxic to the mitotic spindle and is a strong cytostatic agent. Paclitaxel has a good effect on the treatment of ovarian cancer and breast cancer, and has a good effect on the treatment of prostate cancer, upper gastrointestinal cancer, small cell and non-small cell lung cancer. On February 29, 1992, the U.S. Food and Drug Administration officially approved paclitaxel as a new antineoplastic drug for the treatment of advanced ovarian cancer.

As we all know, paclitaxel has great application value and economic benefits, but its wide application has obvious disadvantages: the solubility of paclitaxel in water is 0.006mg/ml, and the key technology of its intravenous infusion preparation is to solve its solubility in water. The preparation product of current clinical application all is to adopt polyoxyethylene castor oil (Cremophor, surfactant) and dehydrated alcohol (v/v1:1) to make solvent, make the solution of 30mg/5ml, add water to dissolve, and in 8 - Stable within 12 hours (no crystallization), dilute with solvent for injection (such as 5% glucose, etc.) in clinical application, and then infuse intravenously. This preparation form has severe defects such as large toxic and side effects and inconvenient application. The preparation of paclitaxel intravenous injection preparations without polyoxyethylene castor oil is a technical problem urgently needed to be solved.

As a new technology for parenteral and enteral administration of insoluble drugs in water, nano-preparation is a hot spot in the research of pharmaceutical preparations. It can improve the dissolution rate of drugs, enhance the targeting, sustained release and low toxicity of drugs. However, nano-preparation is a new form of preparation, the safety of the carrier used, easy selectivity, and process methods are the keys to the success of nano-product development.

US5439686 discloses paclitaxel in the form of suspended granules coated with albumin (as a stabilizer). The protein and paclitaxel in the biocompatible dispersion medium are subjected to high-shear treatment to produce particles with a diameter of less than 1 micron, and can regenerate extremely small particles with a diameter of less than 200nm. This method uses albumin as a stabilizer, which is expensive and has high production costs.

Cyclodextrin (Cyclodextrin, usually abbreviated as CD) is a kind of truncated cone-shaped macrocyclic molecule composed of D-type glucopyranose units connected end to end by α-1,4 glycosidic bonds. It has a hydrophobic cavity and a hydrophilic surface. Therefore, cyclodextrins can selectively bond various inorganic, organic, and biomolecules to form supramolecular complexes. Cyclodextrins customarily use a Greek letter to indicate the number of glucose units, the most common of which are α, β, and γ cyclodextrins, which have 6, 7, and 8 glucose units, respectively. The β-cyclodextrin containing 7 glucose units can be prepared in large quantities by enzymatic degradation of starch, and is cheap, stable and non-toxic. Using cyclodextrin as a parent to construct functional molecules can create good conditions for the inclusion of drug molecules.

Hydroxypropyl-β-cyclodextrin (HP-β-CD) not only has an excellent enveloping effect on many compounds, improves the stability of the enveloped substances, but also has high water solubility and improves the enveloping effect in the organism. drug release rate and bioavailability. On the other hand, HP-β-CD has low hemolytic activity, low surface activity, no irritation to skin and muscle, and is a good co-solvent.

Belgian Sisen Laboratory Co., Ltd. mixed paclitaxel, acetyl-γ-cyclodextrin and hydroxypropyl-β-cyclodextrin in ethanol, and freeze-dried the resulting solid for water solubility research, and has applied for a Chinese patent (Application No. 98811010.5), but does not mention nanoparticles.

Contents of the invention

The object of the present invention is to provide a taxane cyclodextrin inclusion complex nanoparticle. Through the research, it is found that using HP-β-CD as the carrier, inclusion compound nanoparticles obtained by including taxanes can achieve a good solubilization effect. The invention also provides a method for preparing taxane cyclodextrin inclusion compound nanoparticles, and a form of pharmaceutical preparation composed of taxane cyclodextrin inclusion compound nanoparticles.

Hydroxypropyl beta cyclodextrin was selected as the carrier of taxane drugs to prepare taxane-hydroxypropyl beta cyclodextrin inclusion complex nanoparticles with a particle size of 40-200nm. Certain tumor targeting and long circulation. It is rapidly soluble in water, increases the solubility of paclitaxel from 0.006mg/ml to 0.6-3mg/ml, solubilizes by 100-500 times, and has an encapsulation rate of 80-90%. Under normal temperature conditions, it has good stability.

The clathrate nanoparticles of the present invention are prepared by the following method:

1. Prepare an aqueous solution of hydroxypropyl beta cyclodextrin;

2. Add stabilizer, mix well and heat to 30°C-80°C;

3. Add the taxane drug solution under stirring, keep the temperature and stir for 1-6hr;

4. Freeze drying or rotary evaporation to remove the organic solvent.

The taxane cyclodextrin inclusion compound prepared by the present invention can significantly increase the tolerance dose and tumor inhibition rate in the animal drug efficacy test.

The preparation of the present invention adopts conventional auxiliary materials and process conditions, can be produced on an industrial scale and with high efficiency, and has stable product quality. It can be directly or secondary processed into injection or oral preparations. It is a unique, universally applicable, low-cost industrialized Preparation.

In the present invention,

Taxanes include paclitaxel and docetaxel, with paclitaxel being preferred. Pre-dissolved in an organic solvent such as alcohol.

Hydroxypropyl beta cyclodextrin refers to a compound composed of a ring of 7D-glucose units linked by α-1,4 glycosidic bonds, in which a certain proportion of hydroxyl groups are substituted by hydroxypropyl functional groups through ether bonds. Wherein, the molar ratio of hydroxypropyl beta cyclodextrin to taxane is 1-100:1, preferably 10:1.

Prepare hydroxypropyl beta-cyclodextrin aqueous solution by conventional method, add stabilizer, described stabilizer is polyvinylpyrrolidone (povidone) or carboxymethylcellulose sodium (CMC-Na) or hydroxypropyl methyl One or several types of cellulose (HPMC), the amount used is 1-10 times the mass of taxane. In a preferred solution of the present invention, the stabilizer is povidone, especially povidone K29/32.

After the stabilizer is uniformly mixed with the aqueous solution of hydroxypropyl beta cyclodextrin, it is heated to 30°C-80°C, preferably 40°C-70°C.

When adding the taxane solution, the stirring speed is 300-1200 rpm, preferably 900 rpm.

Maintain temperature and stir for 1-6 hr, preferably 3 hr.

To remove organic solvents, freeze drying or rotary evaporation can be utilized. In this process, leavening agents such as mannitol and dextran can be added.

The taxane cyclodextrin inclusion compound of the present invention has significantly improved solubility in the aqueous phase, and therefore, can be delivered to cancer patients in a more efficient manner.

In the present invention, the aqueous solution of the hydroxypropyl-β-cyclodextrin inclusion product of paclitaxel can be dried to form a powder, thereby obtaining the paclitaxel cyclodextrin inclusion compound with high solubility and high stability.

The paclitaxel cyclodextrin inclusion compound of the present invention can be used in various forms. For example, it can be prepared as an injectable drug or used as a powder, and can also be mixed with a pharmaceutical diluent, carrier or excipient to form a pharmaceutical composition. Administration to the patient may be intravenous, oral or by other routes. Through this mode of administration, an effective dose of paclitaxel can be provided to the patient.

In the present invention, the detection method adopted is as follows:

Nanoparticle size: Nicomp380 particle size analyzer Hitachi H120 transmission electron microscope measurement

Solubility: Dissolve a certain amount of clathrate in water, ultrasonically oscillate until no solid dissolves, filter, dry and weigh the obtained solid, and calculate the solubility value of the clathrate after conversion.

Stability: After standing at room temperature for 6-8 hours, detect the inclusion compound particle size.

Animal experiment--

Tolerated dose: give mice 50mg/kg, once a day, for 3 consecutive days. Observe for 1 week. The dose was increased to 75mg/kg, 100mg/kg, and the experiment was repeated. Observe the changes in animal signs and determine the maximum tolerated dose.

Drug efficacy evaluation: Nude mice were subcutaneously inoculated with Lewis lung cancer cells in the right axilla, and after a week, they were injected with paclitaxel-cyclodextrin-coated nanoparticles or commercially available paclitaxel at a dose of 40 mg/kg on the 4th and 8th days. After 2 weeks, observe the change of tumor volume.

Description of drawings

Figure 1 Scanning electron micrograph of paclitaxel cyclodextrin inclusion complex nanoparticles

Figure 2 Nicomp380 particle size analyzer particle size distribution diagram, in which:

99.2% of the particles have an average particle size of 77.9nm

The average particle size of 0.8% of the particles was 327.1 nm.

Figure 3 Comparison of tumor inhibitory effects between paclitaxel cyclodextrin inclusion complex nanoparticles and commercially available paclitaxel

Visual map of tumor-bearing tumor volume in nude mice after 2 weeks of administration of A

The upper left is the control group;

The upper right is the commercial paclitaxel group;

The bottom left is the clathrate nanoparticle group.

B curve of tumor-bearing tumor volume change in nude mice after administration

specific embodiment

Preparation of Paclitaxel Cyclodextrin Nanoparticles--

The prescription is as follows:

Paclitaxel 2mg

Hydroxypropyl Beta Cyclodextrin 15mg

Povidone K29/32 10mg

Method: Add povidone K29/32 to the hydroxypropyl beta-cyclodextrin solution, mix well and heat to 40°C, add paclitaxel drug solution under magnetic stirring at 900rpm, keep the temperature and stir for 3hr, freeze-dry (add nectar Alcohol, dextran and other leavening agents) or rotary evaporation to remove the organic solvent, to obtain freeze-dried powder injection.

After testing, more than 90% of the obtained paclitaxel nanoparticles have a particle diameter of 70-80nm, a solubility of paclitaxel of 0.6 mg/ml, and can be stable for 6-8 hours at room temperature.

After dissolving the obtained freeze-dried powder injection in water, give 40 mg/kg to tumor-bearing nude mice. After 2 weeks, the tumor inhibition rate of the paclitaxel nanoparticle group was measured to be 99.85%, and the tumor inhibition rate of the commercially available group was 85.79%, which was significant. difference.

Claims (2)

1. a clathrate is made up of taxane, HYDROXYPROPYL BETA-CYCLODEXTRIN and stabilizing agent; Described taxane is a paclitaxel, and described stabilizing agent is a polyvidone, it is characterized in that, the mol ratio of described taxane and HYDROXYPROPYL BETA-CYCLODEXTRIN is 1:1-10, and the consumption of described stabilizing agent is 1-10 a times of taxane quality, and the particle diameter of clathrate is 40-200nm.

2. a kind of pharmaceutical composition that comprises the described clathrate of claim 1.

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