CN101632637B - A kind of docetaxel lipid preparation and preparation method thereof - Google Patents

Abstract

The invention discloses a polyene taxol lipid preparation and a method for preparing the same. The polyene taxol lipid preparation comprises the following components of polyene taxol, polyethylene glycol-12-hydroxyl stearate and phospholipid. The polyene taxol lipid preparation improves the safety and the compliance of the polyene taxol preparation; and after being diluted, the polyene taxol lipid preparation can be automatically emulsified into nano particles for intravenous injection so as to enhance the in vivo absorption of the polyene taxol and fulfill the tumor targeting aim. Moreover, the method has the advantages of simple and convenient preparation process and the suitability for industrialized production.

Description

A kind of docetaxel lipid preparation and preparation method thereof

technical field

The invention relates to the technical field of medicine, in particular to a docetaxel lipid preparation and a preparation method thereof.

Background technique

Docetaxel (Docetaxel, ) is a semi-synthetic anticancer drug and one of the taxane anticancer drugs approved by the FDA. Docetaxel is produced semi-synthetically from the inactive precursor (diterpene taxane 10-deacetylbaccatin III) extracted from the needles of the European yew tree, and its chemical structure is similar to that of paclitaxel (Paclitaxel, ) compared with the existence of two changes, one is to replace the acetyl group with a carboxyl group at the C-10 position of the taxol B ring, and the other is a change in the C-13 side chain (for example, on the side chain of taxol with N-tert-butoxycarbonyl instead of N-benzoyl). The difference in chemical structure leads to different activities of docetaxel and paclitaxel, and its cytotoxicity is 1.3-12 times that of paclitaxel.

Docetaxel is a microtubule depolymerization inhibitor, which acts on the microtubule/tubulin system, promotes the assembly of microtubule dimers into microtubules, and prevents the process of depolymerization to stabilize microtubules, allowing cells to undergo G2/ M phase arrest, thereby inhibiting the mitosis and proliferation of cancer cells, this mechanism of action results in it having less serious side effects than paclitaxel. Clinical studies have shown that docetaxel has good anticancer activity on various tumor diseases, and has good effects on the treatment of breast cancer, non-small cell lung cancer, pancreatic cancer, soft tissue sarcoma, head and neck cancer, gastric cancer, ovarian cancer and prostate cancer. curative effect.

由法国安万特公司开发，1996年获得FDA批准，用于蒽环类药物化疗失败后的局部晚期或转移性乳腺癌，1999年被批准局部晚期或转移性非小细胞肺癌(NSCLC)的二线应用，2002年11月被FDA批准与顺铂联合使用，用于不能切除的、局部晚期或转移性NSCLC患者(之前未接受化疗)的治疗。之后，

被批准与泼尼松联合应用于非雄激素依赖的(耐激素的)转移性前列腺癌患者的治疗，与多柔比星和环磷酰胺联用也被批准用于可手术的、结节阳性(node-positive)乳腺癌患者的辅助治疗。Docetaxel

Developed by Aventis, France, it was approved by the FDA in 1996 for locally advanced or metastatic breast cancer after failure of anthracycline chemotherapy, and was approved as a second-line treatment for locally advanced or metastatic non-small cell lung cancer (NSCLC) in 1999 Application, in November 2002, was approved by the FDA in combination with cisplatin for the treatment of unresectable, locally advanced or metastatic NSCLC patients (not previously treated with chemotherapy). after,

Approved in combination with prednisone for the treatment of patients with androgen-independent (hormone-resistant) metastatic prostate cancer and in combination with doxorubicin and cyclophosphamide for operable, nodule-positive Adjuvant therapy for (node-positive) breast cancer patients.

采用聚山梨醇酯80(吐温80)/乙醇作为溶媒，其规格为含有20mg(0.5ml)或80mg(2.0ml)多烯紫杉醇(无水的)单次剂量小瓶，每ml含有40mg的多烯紫杉醇(无水的)和1040mg的吐温80。要求使用前稀释，因此配有无菌无热原的单次剂量稀释液，为含有13％乙醇的注射用水。Due to the poor water solubility of docetaxel, freeze-dried powder injection and water injection are mainly used clinically. Docetaxel

Using polysorbate 80 (Tween 80)/ethanol as the vehicle, its specification is a single-dose vial containing 20 mg (0.5 ml) or 80 mg (2.0 ml) of docetaxel (anhydrous), and each ml contains 40 mg of docetaxel Paclitaxel (anhydrous) and Tween 80 at 1040 mg. Dilution prior to use is required, so a sterile pyrogen-free single-dose diluent is provided as Water for Injection containing 13% ethanol.

Tween 80 and ethanol are mainly used to increase the solubility of docetaxel, but the presence of this solvent can cause hemolysis and severe allergic reactions, such as hypotension and/or bronchospasm and/or generalized rash/erythema. In order to reduce the incidence and severity of allergic reactions, steroids and other histamine blocking drugs need to be pre-administered clinically, but this pre-administration method also has some adverse reactions, such as Cushing's syndrome, infectious complications , hyperglycemia, hypertension, and psychotic effects (including steroid-induced psychosis), etc., especially during long-term administration, solvents also promote the dissolution of plasticizers from polyoxyethylene (PVC) bags and tubing, and may produce Other adverse effects associated with these agents, such as neurological disorders and tumor cell resistance. Adverse reactions limit the clinical application of this drug. Therefore, it is necessary to improve the solubility of docetaxel through new drug delivery systems or preparations, further improve its bioavailability, and reduce its toxicity.

Contents of the invention

The object of the present invention is to provide a docetaxel lipid preparation and a preparation method thereof.

The docetaxel lipid preparation provided by the invention comprises the following components: docetaxel, polyethylene glycol-dodecyl hydroxystearate and phospholipids; the content of phosphatidylcholine in the phospholipids is greater than 80%.

The preparation also includes a co-solvent; the co-solvent is a monohydric alcohol, a dihydric alcohol or a trihydric alcohol with a total carbon number of three or less.

The co-solvent may specifically be at least one of absolute ethanol, propylene glycol, and glycerin.

A pH adjusting agent may also be included in the formulation.

The pH regulator can specifically be anhydrous citric acid.

The preparation may include the following components in parts by mass: 0.1-5 docetaxel, 15-50 polyethylene glycol-lauryl hydroxystearate, 5-20 phospholipids, 30-80 cosolvents, anhydrous lemon Acid 0.1-0.2.

The preparation may specifically include the following components in parts by mass: 1 docetaxel, 15-50 polyethylene glycol-lauryl hydroxystearate, 5-20 phospholipids, 30-80 co-solvents, anhydrous citric acid 0.18.

Certainly, the said preparation can also only consist of the following components by mass: 1 docetaxel, 15-50 polyethylene glycol-lauryl hydroxystearate, 5-20 phospholipids, 30-80 cosolvents , anhydrous citric acid 0.18.

The above-mentioned docetaxel lipid preparation is a light yellow oily liquid with good stability, and can spontaneously emulsify after dilution to form nanoparticles with a particle diameter in the range of 10-200nm.

The present invention also provides a method for preparing the docetaxel lipid preparation, comprising the steps of:

1) adding the phospholipid into a co-solvent, heating and dissolving at 45-55° C. to obtain solution A; adding docetaxel, anhydrous citric acid and polyethylene glycol-lauryl hydroxystearate into a co-solvent, and dissolving at room temperature, Obtain solution B;

2) Mix solution A and solution B.

The present invention also provides a docetaxel nano-preparation, which is obtained by diluting the docetaxel lipid preparation 5-100 times, preferably 20 times, with physiological saline solution, glucose solution or water for injection.

The above-mentioned docetaxel nano-preparation can be directly used as an injection.

The preparation method of the above docetaxel nano-preparation also belongs to the protection scope of the present invention.

HS15(聚乙二醇-十二羟基硬脂酸酯)，制成一种新的脂质制剂，可以降低毒性及过敏反应，同时该制剂在临用时用生理盐水溶液、葡萄糖水溶液、或注射用水稀释后可自发乳化成纳米级的粒子，吸收效果大大增强，从而药效得以进一步提高。The inventor aimed at the problems existing in the clinical application of docetaxel, using high-purity phospholipids and a new solubilizer

HS15 (polyethylene glycol-dodecyl hydroxystearate), made into a new lipid preparation, can reduce toxicity and allergic reactions. After dilution, it can be spontaneously emulsified into nano-sized particles, and the absorption effect is greatly enhanced, so that the drug effect can be further improved.

HS15，是一种新型的高性能、极低毒性的增溶剂，其优越性体现在：

HS15 is a new type of solubilizer with high performance and extremely low toxicity. Its advantages are reflected in:

HS15的使用，不同药物如克霉唑、雌二醇、磺胺噻唑、心痛定、酰胺咪嗪、吡罗昔康等的水溶性增加，根据分子结构不同其水溶解度被提高10-100因数，不管化学结构或药物溶解的最大剂量怎样，胶粒的直径不变，均在13nm左右。

HS15的这种高增溶能力使低容量高剂量的注射成为可能。① High solubilizing ability: the comparative study with Tween 80 shows that, The solubility of HS15 for hydrophobic drugs increased linearly with increasing solvent concentration, while using Tween 80, the amount of drug dissolved did not increase further at higher concentrations. While another experiment showed that with 20% of

The use of HS15 increases the water solubility of different drugs such as clotrimazole, estradiol, sulfathiazole, nifedipine, carbamazepine, piroxicam, etc., and their water solubility is increased by a factor of 10-100 depending on the molecular structure, regardless of the chemical structure Or what about the maximum dose of drug dissolution, the diameter of the colloidal particles remains unchanged, all around 13nm.

This high solubilizing ability of HS15 enables low-volume, high-dose injections.

HS15组仅为220nMol；60min后吐温80组的血清组胺水平为247nMol，而HS15组仅为8nMol。②Low histamine release: No need to use antihistamines and corticosteroids before use. Intravenous injection compared to Tween 80 After HS15, the serum histamine level of the dogs was many times lower, and the serum histamine level of the Tween 80 group was greater than 50000nMol after 15min, while

HS15 group was only 220nMol; after 60min, the serum histamine level of Tween 80 group was 247nMol, while The HS15 group was only 8nMol.

HS15组10倍以上，表明这种新的增溶剂具有较低的毒性和刺激性。③Low hemolysis: The comparative study with Tween 80 showed that although the hemolysis was enhanced with the increase of the solubilizer concentration, the amount of erythrocyte lysis in the Tween 80 group was higher than that in the Tween 80 group.

HS15 group more than 10 times, indicating that this new solubilizer has lower toxicity and irritation.

In addition, this new solubilizer has high physiological tolerance, low viscosity, and can be steam sterilized without expensive aseptic manufacturing processes. The solvent is currently included in the German Pharmacopoeia, and is available in the United States and Canada. Approved for human injection.

HS15以及合适的载体材料，大大提高了多烯紫杉醇的溶解性，同时可避免使用吐温80带来的毒副作用，提高了多烯紫杉醇制剂的安全性和顺应性；而且该制剂稀释后自发乳化形成纳米粒子，用于静脉注射，可以增加多烯紫杉醇的体内吸收，并且通过控制其乳化粒度的大小可达到肿瘤靶向的目的；本发明的制备工艺简单易行，适合工业化生产。The docetaxel lipid preparation provided by the invention is due to having selected novel solubilizing agent

HS15 and suitable carrier materials greatly improve the solubility of docetaxel, avoid the toxic and side effects caused by the use of Tween 80, and improve the safety and compliance of docetaxel preparations; moreover, the preparations emulsify spontaneously after dilution Forming nanoparticles for intravenous injection can increase the absorption of docetaxel in vivo, and can achieve the purpose of tumor targeting by controlling the emulsified particle size; the preparation process of the invention is simple and easy, and is suitable for industrial production.

The following examples facilitate a better understanding of the present invention, but do not limit the present invention.

Description of drawings

Figure 1 is the result of particle size detection after emulsification of docetaxel nano-preparation - multimodal distribution of light intensity particle size

Figure 2 is the result of particle size detection after emulsification of docetaxel nano-preparation - normal distribution of light intensity particle size

Detailed ways

The experimental methods in the following examples are conventional methods unless otherwise specified.

Embodiment 1, the preparation of docetaxel lipid preparation

One, the preparation of docetaxel lipid preparation

1. Weigh 5g of phospholipid (Germany Lipoid Company) and 10g of absolute ethanol, heat and dissolve in a water bath at 45°C to obtain solution A.

2. Weigh 1g docetaxel, 15g HS15 (German BASF company), 0.18g citric acid, 20g glycerol, 50g absolute ethanol, dissolve at room temperature to obtain solution B.

3. Mix solution A and solution B, stir at room temperature, mix evenly, and filter to obtain docetaxel lipid preparation, which is protected by nitrogen gas and sealed for storage.

2. Preparation of Docetaxel Nano-Preparation

Dilute the docetaxel lipid preparation prepared in step 1 with 20 times the volume of glucose solution to obtain the docetaxel nano-preparation.

3. Determination of particle size of docetaxel nano-preparation

The particle size of docetaxel nano-preparation was detected by ZetaPALS laser dynamic light scattering instrument from Brookhaven Instrument Company, USA. The average particle diameter of the docetaxel nano-preparation is 76nm. After 6 months, the particle size of the docetaxel nano-preparation was detected again, and the average particle size was 74nm, with no significant change.

Embodiment 2, the preparation of docetaxel lipid preparation

One, the preparation of docetaxel lipid preparation

1. Weigh 8g of phospholipids (Germany Lipoid Company) and 12g of propylene glycol, heat and dissolve in a water bath at 55°C to obtain solution A.

2. Weigh 1g docetaxel, 30g HS15 (German BASF company), 0.18g citric acid, 50g absolute ethanol, and dissolve at room temperature to obtain solution B.

3. Mix solution A and solution B, stir at room temperature, mix evenly, and filter to obtain docetaxel lipid preparation, which is protected by nitrogen gas and sealed for storage.

2. Preparation of Docetaxel Nano-Preparation

The docetaxel lipid preparation prepared in step 1 was diluted with 20 times the volume of physiological saline to obtain the docetaxel nano-preparation.

3. Determination of particle size of docetaxel nano-preparation

The particle size of docetaxel nano-preparation was detected by ZetaPALS laser dynamic light scattering instrument from Brookhaven Instrument Company, USA. In a stable environment of 25±0.1°C, the particle size detection results are shown in Figure 1 and Figure 2. The results showed that the average particle diameter of docetaxel nano-preparation was 18.4nm. After 6 months, the particle size of the docetaxel nano-preparation was detected again, and the average particle size was 23.8nm, with no significant change.

Embodiment 3, the preparation of docetaxel lipid preparation

One, the preparation of docetaxel lipid preparation

1. Weigh 12g of phospholipid (Germany Lipoid Company) and 12g of propylene glycol, and heat and dissolve in a water bath at 50°C to obtain solution A.

2. Weigh 1g docetaxel, 36g HS15 (German BASF company), 6g glycerin, 0.18g citric acid, 34g absolute ethanol, and dissolve at room temperature to obtain solution B.

3. Mix solution A and solution B, stir at room temperature, mix evenly, and filter to obtain docetaxel lipid preparation, which is protected by nitrogen gas and sealed for storage.

2. Preparation of Docetaxel Nano-Preparation

Dilute the docetaxel lipid formulation prepared in step 1 with 20 times the volume of water for injection to obtain the docetaxel nano-preparation.

3. Determination of particle size of docetaxel nano-preparation

The particle size of docetaxel nano-preparation was detected by ZetaPALS laser dynamic light scattering instrument from Brookhaven Instrument Company, USA. The average particle size of the docetaxel nano-preparation is 126.9nm. After 6 months, the particle size of the docetaxel nano-preparation was detected again, and the average particle size was 101.9nm, with no significant change.

Embodiment 4, the preparation of docetaxel lipid preparation

One, the preparation of docetaxel lipid preparation

1. Weigh 20g of phospholipid (Germany Lipoid Company) and 10g of absolute ethanol, heat and dissolve in a water bath at 45°C to obtain solution A.

2. Weigh 1g docetaxel, 50g HS15 (German BASF company), 0.18g citric acid, 20g absolute ethanol, and dissolve at room temperature to obtain solution B.

3. Mix solution A and solution B, stir at room temperature, mix evenly, and filter to obtain docetaxel lipid preparation, which is protected by nitrogen gas and sealed for storage.

2. Preparation of Docetaxel Nano-Preparation

Dilute the docetaxel lipid preparation prepared in step 1 with 20 times the volume of glucose solution to obtain the docetaxel nano-preparation.

3. Determination of particle size of docetaxel nano-preparation

The particle size of docetaxel nano-preparation was detected by ZetaPALS laser dynamic light scattering instrument from Brookhaven Instrument Company, USA. The average particle size of the docetaxel nano-preparation is 178nm. After 6 months, the particle size of the docetaxel nano-preparation was detected again, and the average particle size was 181nm, with no significant change.

Embodiment 5, acute toxicity test

1. Experimental animals and grouping

100 healthy Kunming mice (18-22g), half male and half male. The mice were randomly divided into 10 groups, 10 mice in each group.

2. Preparation of Docetaxel Injection

Prepare a sample containing 40 mg docetaxel and 1040 mg Tween 80 per ml according to the specifications of commercially available docetaxel injection, and dilute it 4 times with water for injection containing 13% ethanol to obtain docetaxel injection as a control group. .

3. Administration

Control group I (10 mice): intraperitoneal injection of docetaxel injection at a dose of 100 mg/Kg.

Control group II (10 mice): intraperitoneal injection of docetaxel injection at a dose of 150 mg/Kg.

Experimental group I (10 mice): the docetaxel lipid preparation prepared in Example 1 was intraperitoneally injected with a dosage of 100 mg/Kg.

Experimental group II (10 mice): the docetaxel lipid preparation prepared in Example 1 was injected intraperitoneally, and the administration dose was 150 mg/Kg.

Experimental group III (10 mice): the docetaxel lipid preparation prepared in Example 2 was injected intraperitoneally, and the dosage was 100 mg/Kg.

Experimental group IV (10 mice): intraperitoneal injection of the docetaxel lipid preparation prepared in Example 2, with a dosage of 150 mg/Kg.

Experimental group V (10 mice): the docetaxel lipid preparation prepared in Example 3 was intraperitoneally injected with a dose of 100 mg/Kg.

Experimental group VI (10 mice): the docetaxel lipid preparation prepared in Example 3 was injected intraperitoneally, and the dosage was 150 mg/Kg.

Experimental group VII (10 mice): the docetaxel lipid preparation prepared in Example 4 was injected intraperitoneally, and the dosage was 100 mg/Kg.

Experimental group VIII (10 mice): the docetaxel lipid preparation prepared in Example 4 was injected intraperitoneally, and the dosage was 150 mg/Kg.

Observe the symptoms of toxic reaction in the experimental group and the control group mice after administration.

The results showed that: when the administration dose was 100 mg/kg, the mice in the control group showed obvious symptoms of toxic reaction, with limbs lying on the ground, shortness of breath, and listless state. The mice died in 3 days after the administration, and the death rate of the mice after one week was 20%; while the 4 groups of mice injected with docetaxel lipid preparations had mild symptoms of toxicity, such as slow response, lethargy, and weight loss, and no mice died one week after administration.

When the administration dose was 150 mg/kg, mice in the control group died 1 day after administration, and the death rate of the mice was 70% after one week; while the toxicity symptoms of mice in the 4 groups injected with docetaxel lipid preparations were alleviated , Experimental groups II, IV, VI, and VIII began to have mice death on the 2nd day, the 3rd day, the 2nd day, and the 3rd day after the administration, and the mouse death rate was 40%, 50% and 50% respectively in one week after the administration. %, 50%, 40%, lower than the control group.

The above experimental results show that compared with docetaxel injection, the toxicity of docetaxel lipid preparation is reduced.

Claims (4)

1. A docetaxel lipid preparation is prepared by the following method: 1) Weigh 12g of phospholipid and 12g of propylene glycol, heat and dissolve in a water bath at 50°C to obtain solution A; 2) Weigh 1g of docetaxel, 36g of polyethylene glycol-lauryl hydroxystearate, 6g of glycerin, 0.18g of citric acid and 34g of absolute ethanol, and dissolve at room temperature to obtain solution B; 3) Mix solution A and solution B, stir at room temperature, mix evenly, and filter to obtain docetaxel lipid preparation, which is protected by nitrogen gas and sealed for storage. 2. a method for preparing the docetaxel lipid preparation claimed in claim 1, its steps are as follows: 1) Weigh 12g of phospholipid and 12g of propylene glycol, heat and dissolve in a water bath at 50°C to obtain solution A; 2) Weigh 1g of docetaxel, 36g of polyethylene glycol-lauryl hydroxystearate, 6g of glycerin, 0.18g of citric acid and 34g of absolute ethanol, and dissolve at room temperature to obtain solution B; 3) Mix solution A and solution B, stir at room temperature, mix evenly, and filter to obtain docetaxel lipid preparation, which is protected by nitrogen gas and sealed for storage. 3. A method for preparing a docetaxel nano-preparation, comprising diluting the docetaxel lipid preparation of claim 1 with 20 times the volume of water for injection to obtain a docetaxel nano-preparation. 4. A docetaxel nano-preparation prepared by the method according to claim 3.

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