WO2023030474A1 - Hydrophobic drug polymer micelle and preparation method therefor - Google Patents

Abstract

Provided is a polyethylene glycol monomethyl ether-polylactic acid block copolymer and a preparation method therefor. The polyethylene glycol monomethyl ether-polylactic acid block copolymer is a block copolymer formed by means of ring-opening polymerization of D,L-lactide and polyethylene glycol monomethyl ether, wherein the feed mass ratio of polyethylene glycol monomethyl ether to D,L-lactide is 1:1.05-1.25. Also provided is a hydrophobic drug polymer micelle lyophilized preparation. The lyophilized preparation contains a hydrophobic drug and the polyethylene glycol monomethyl ether-polylactic acid block copolymer. The hydrophobic drug polymer micelle lyophilized preparation has higher purity and redissolution stability, and is more suitable for clinical use.

Description

A kind of hydrophobic drug polymer micelle and preparation method thereof

This disclosure claims the priority of the Chinese patent application with the patent application number 202111033302.5 and the invention title "a hydrophobic drug polymer micelle and its preparation method" submitted to the State Intellectual Property Office of China on September 3, 2021. The entirety of the aforementioned prior application is incorporated by reference into this disclosure.

technical field

The present disclosure relates to the field of medicine, in particular, to a hydrophobic drug polymer micelle freeze-dried preparation and a preparation method thereof.

Background technique

Taxanes are a large class of natural products obtained from plants, and are an important class of anticancer drugs used clinically. Due to the very low solubility of taxanes in water, their clinical application is greatly limited. Docetaxel (DTX), also known as docetaxel, has a molecular formula of C ₄₃ H ₅₃ NO ₁₄ and a molecular weight of 807.88. It is a taxane antineoplastic drug, which can bind to free tubulin and promote microtubule The protein assembles into stable microtubules, and at the same time inhibits their depolymerization, leading to the production of microtubules that lose their normal functions and the fixation of microtubules, thereby inhibiting cell mitosis and exerting anti-tumor effects. However, docetaxel is difficult to absorb when taken orally, and has disadvantages such as poor water solubility, short half-life, and high toxicity. Clinically, it is usually administered by intravenous infusion. At present, docetaxel injections sold domestically and abroad are made by dissolving docetaxel in Tween-80. In clinical application, it needs to be prepared with a special solvent and diluted with an isotonic solution, which is cumbersome to use and requires strict operation. The preparation contains a large amount of Tween-80, which is likely to cause adverse reactions such as hemolysis and allergy, and it is necessary to take dexamethasone and other drugs in advance for prevention and treatment, which is inconvenient for clinical use and has low safety. Paclitaxel is a taxanol antineoplastic drug extracted from yew or yew tree. It is a white crystalline powder, insoluble in water, and easily soluble in organic solvents such as chloroform and acetone. Paclitaxel injection currently used clinically contains polyoxyethylene-modified castor oil and absolute ethanol, and polyoxyethylene-modified castor oil and ethanol often cause strong side effects, such as hypersensitivity, hemolysis, kidney Toxicity, neurotoxicity and cardiotoxicity, etc., may lead to death in severe cases.

Nanopolymer micelle is a drug-loading system developed in recent years for insoluble drugs. It has a core-shell structure, in which the core is the hydrophobic part and the shell is the hydrophilic part. Polymer micelles can encapsulate poorly soluble drugs in the core part to solubilize poorly soluble drugs.

CN201410326208.2 discloses a docetaxel nano-polymer micelle freeze-dried preparation and its preparation method, which is prepared by using polyethylene glycol monomethyl ether and D, L-lactide with a mass ratio of 1:0.99 Polyethylene glycol monomethyl ether-polylactic acid block copolymer encapsulated docetaxel. The present disclosure is further optimized on the basis of CN201410326208.2, and provides a hydrophobic drug polymer micelle freeze-dried preparation with higher purity and reconstitution stability.

Contents of the invention

The purpose of the present disclosure is to provide a lyophilized preparation of hydrophobic drug polymer micelles with higher purity and reconstitution stability.

Another object of the present disclosure is to provide a preparation method of the hydrophobic drug polymer micelle freeze-dried preparation.

Another object of the present disclosure is to provide a polyethylene glycol monomethyl ether-polylactic acid block copolymer.

The disclosure provides a polyethylene glycol monomethyl ether-polylactic acid block copolymer, and the polyethylene glycol monomethyl ether-polylactic acid block copolymer is D,L-lactide and polyethylene glycol A block copolymer formed by the polymerization of monomethyl ether, wherein the mass ratio of polyethylene glycol monomethyl ether to D,L-lactide is 1:1.05-1.25, and the polyethylene glycol monomethyl ether- The polylactic acid block copolymer is prepared by the following method: (a) adding polyethylene glycol monomethyl ether into the reactor, heating and melting under vacuum, and replacing with inert gas; (b) adding D,L-lactide, inert Gas replacement, adding a metal catalyst, ensuring negative pressure or inert gas protection in the reactor, and then heating to 125-150°C for reaction; (c) cooling down after the reaction, adding the first organic solvent to dissolve, adding the second organic solvent, and filtering , and vacuum-dry the filter cake to obtain polyethylene glycol monomethyl ether-polylactic acid block copolymer.

In some embodiments, the mass ratio of polyethylene glycol monomethyl ether to D,L-lactide is 1:1.1-1.2.

In some embodiments, the mass ratio of polyethylene glycol monomethyl ether to D,L-lactide is 1:1.1.

In some embodiments, the polyethylene glycol monomethyl ether in step (a) has a molecular weight of 1000-20000, preferably 1000-5000, more preferably 1800-2200, even more preferably 2000.

In some embodiments, the polyethylene glycol monomethyl ether in step (a) is heated to 60-130° C. under vacuum.

In some embodiments, the metal catalyst of step (b) is stannous isooctanoate.

In some embodiments, the mass of stannous isooctanoate accounts for 0.05-0.5 wt%, preferably 0.15-0.3 wt%, of the total mass of D,L-lactide and polyethylene glycol monomethyl ether.

In some embodiments, the reaction time of step (b) is 1-20 h, preferably 3-10 h, more preferably 4-6 h.

In some embodiments, the first organic solvent described in step (c) is one of acetonitrile, acetone, dichloromethane, dimethylformamide, dimethylsulfoxide, tetrahydrofuran, short-chain fatty alcohol or ethyl acetate one or more, preferably one or more of dichloromethane, chloroform, methyl alcohol, ethanol, isopropanol, more preferably dichloromethane; the consumption of the first organic solvent accounts for the quality of polyethylene glycol monomethyl ether 0.1-100 times the volume (L/kg), preferably 0.2-50 times the volume (L/kg), more preferably 0.5-20 times the volume (L/kg).

In some embodiments, the second organic solvent described in step (c) is one or more of diethyl ether, isopropyl ether, methyl tert-butyl ether, n-heptane, cyclohexane, petroleum ether, preferably One or more of diethyl ether, isopropyl ether or methyl tert-butyl ether, more preferably diethyl ether; the amount of the second organic solvent accounts for 1 to 100 times the volume (L/kg) of the total mass of polyethylene glycol monomethyl ether , preferably 10 to 50 times the volume (L/kg), more preferably 15 to 20 times the volume (L/kg).

In some embodiments, after the reaction in step (b), it also includes the steps of adding a third organic solvent and activated carbon, stirring, filtering, and concentrating the filtrate under reduced pressure.

In some embodiments, the third organic solvent is dichloromethane, chloroform, ethyl acetate, isopropyl acetate, butyl acetate, butanone, toluene, acetone, acetonitrile, dimethylformamide, di One or more of methyl sulfoxide, tetrahydrofuran, 1,4-dioxane, and short-chain fatty alcohols, preferably one or more of methylene chloride, chloroform, methanol, ethanol, and isopropanol , more preferably dichloromethane.

In some embodiments, the amount of the third organic solvent accounts for 0.1 to 100 times the volume (L/kg) of polyethylene glycol monomethyl ether, preferably 0.2 to 50 times the volume (L/kg), more preferably 0.5 to 100 times the volume (L/kg). 20 times the volume (L/kg).

In some embodiments, the amount of activated carbon accounts for 1-500wt% of the total mass of D,L-lactide and polyethylene glycol monomethyl ether, preferably 5-200wt%, more preferably 5-75wt%, even more preferably 20-50wt%.

In some embodiments, the metal tin ion content in the polyethylene glycol monomethyl ether-polylactic acid block copolymer is less than 10 ppm, preferably less than 1 ppm, more preferably less than 0.1 ppm.

In some embodiments, the polyethylene glycol monomethyl ether-polylactic acid block copolymer is prepared by the following method: (a) adding polyethylene glycol monomethyl ether into the reactor, filling with nitrogen, and vacuumizing , heating and vacuum drying, inert gas replacement; (b) Add D,L-lactide, fill with nitrogen, vacuumize, keep vacuum in the reactor, inert gas replacement, add metal catalyst under nitrogen protection, nitrogen replacement, to ensure the reaction The container is protected by negative pressure or inert gas, and then heated to 125-150°C to react; (c) After the reaction is completed, the temperature is lowered, the first organic solvent is added to dissolve, the second organic solvent is added, filtered, and the filter cake is vacuum-dried to obtain polyethylene Glycol monomethyl ether-polylactic acid block copolymer.

Another aspect of the present disclosure provides a lyophilized preparation of hydrophobic drug polymer micelles, the lyophilized preparation comprising a hydrophobic drug and the above polyethylene glycol monomethyl ether-polylactic acid block copolymer.

In some embodiments, the mass ratio of the hydrophobic drug to the polyethylene glycol monomethyl ether-polylactic acid block copolymer is 0.01-0.15:1, preferably 0.02-0.12:1, more preferably 0.03-0.10:1 , more preferably 0.04:0.96.

In some embodiments, the lyophilized formulation further comprises a stabilizer.

In some embodiments, the stabilizer is one or more of citric acid, hydrochloric acid, sorbic acid, lactic acid, tartaric acid, malic acid, phosphoric acid, acetic acid, adipic acid, fumaric acid, preferably citric acid One or more of acid, tartaric acid, fumaric acid, more preferably citric acid.

In some embodiments, the mass ratio of the hydrophobic drug to the stabilizer is 1:0.00125-0.25, preferably 1:0.005-0.25, more preferably 1:0.0125-0.125.

In some embodiments, the hydrophobic drug is selected from taxanes.

In some embodiments, the hydrophobic drug is selected from docetaxel or paclitaxel.

In some embodiments, the hydrophobic drug is selected from docetaxel.

In some embodiments, the lyophilized formulation comprises the following ingredients:

The present disclosure also provides a preparation method of the above-mentioned hydrophobic drug polymer micelles lyophilized preparation, comprising the following steps:

(1) Dissolve the above-mentioned polyethylene glycol monomethyl ether-polylactic acid block copolymer and hydrophobic drug in an organic solvent, and remove the organic solvent by rotary evaporation to obtain a gel-like drug film, and then add water to the drug film to dissolve and disperse Described drug film, makes micellar solution;

(2) The micellar solution prepared in step (1) is sterilized by filtration and freeze-dried to obtain a freeze-dried preparation of hydrophobic drug polymer micelles.

In some embodiments, step (1) is to dissolve the above-mentioned polyethylene glycol monomethyl ether-polylactic acid block copolymer, stabilizer and hydrophobic drug in an organic solvent, and remove the organic solvent by rotary evaporation to obtain a gel-like drug film, and then add water to the drug film to dissolve and disperse the drug film to obtain a micellar solution.

In some embodiments, the organic solvent described in step (1) is selected from any of acetonitrile, acetone, dichloromethane, dimethylformamide, dimethyl sulfoxide, tetrahydrofuran, short-chain fatty alcohols, and ethyl acetate. One or more, preferably any one or more of methanol, ethanol, isopropanol, acetonitrile, ethyl acetate, more preferably methanol.

In some embodiments, the amount of the organic solvent described in step (1) is 0.5-12 ml of organic solvent per gram of polyethylene glycol monomethyl ether polylactic acid block copolymer, preferably 3-12 ml of organic solvent.

In some embodiments, in step (1), the amount of water added to the drug film is 2 to 40 ml of water per gram of polyethylene glycol monomethyl ether polylactic acid block copolymer, preferably per gram of polyethylene glycol monomethyl ether polylactic acid block copolymer. Add 10-20ml of water to the alcohol monomethyl ether polylactic acid block copolymer, more preferably add 10-15ml water per gram of polyethylene glycol monomethyl ether polylactic acid block copolymer.

In some embodiments, in step (2), the conditions for removing the organic solvent by rotary evaporation are as follows: a rotation speed of 10-150 rpm, a temperature of 20-80° C., and a time of 1-4 hours.

Another aspect of the present disclosure provides the use of the above-mentioned hydrophobic drug polymer micelle freeze-dried preparation in the preparation of a drug for preventing or treating cancer.

The hydrophobic drug polymer micelle freeze-dried preparation provided by the present disclosure has higher purity and reconstitution stability, and is more suitable for clinical use.

Definitions and Explanations of Terms

Unless otherwise defined in the present disclosure, scientific and technical terms related to the present disclosure shall have the meanings understood by those of ordinary skill in the art.

Polyethylene glycol monomethyl ether can be abbreviated as mPEG or MPEG.

Polyethylene glycol monomethyl ether-polylactic acid block copolymer, also known as polyethylene glycol monomethyl ether-polylactide block copolymer, can be abbreviated as MPEG-PDLLA or mPEG-PDLLA.

Metallic tin ions refer to stannous ions or tin ions, or a mixture of both.

The short-chain fatty alcohol refers to a fatty alcohol having 1 to 6 carbon atoms. It can be monohydric alcohol, dihydric alcohol or polyhydric alcohol. Including but not limited to: methanol, ethanol, isopropanol, n-propanol, n-butanol, isobutanol, n-pentanol, isoamyl alcohol, n-hexanol, isohexanol, 1,2-propanediol, glycerol or 1 , 3-butanediol. Preference is given to methanol, ethanol or isopropanol.

Description of drawings

Figure 1 is the ¹ HNMR spectrum of polyethylene glycol monomethyl ether-polylactic acid block copolymer.

Fig. 2 is a transmission electron micrograph of the freeze-dried preparation of docetaxel polymer micelles.

Detailed ways

The present disclosure will be further described below in conjunction with specific embodiments, and the advantages and characteristics of the present disclosure will become clearer along with the description. Those who do not indicate the specific conditions in the examples are carried out according to the conventional conditions or the conditions suggested by the manufacturer. The reagents or instruments used were not indicated by the manufacturer, and they were all conventional products that could be purchased from the market.

The embodiments of the present disclosure are exemplary only, and do not constitute any limitation to the scope of the present disclosure. Those skilled in the art should understand that without departing from the spirit and scope of the present disclosure, the details and forms of the technical solution of the present disclosure can be modified or replaced, but these modifications and replacements all fall within the protection scope of the present disclosure.

The examples of the present disclosure investigated the molecular weight and distribution of polyethylene glycol monomethyl ether-polylactic acid block copolymer, and the related substances, particle size and span of the docetaxel polymer micelle freeze-dried preparation according to the following methods:

(1) Molecular weight and distribution detection method

¹ HNMR takes 0.5-1.0mL of 1% tetramethylsilane deuterated chloroform 10%-20% (g/mL) solution of this product, scans from 0ppm to 10ppm, and quantifies by direct comparison method. The specific method is: at 3.6ppm It is the peak of CH in polylactic acid, and the peak of CH in polyethylene glycol monomethyl ether is at 5.1ppm. There is the following relationship between the peak area and the number of hydrogen: polyethylene glycol monomethyl ether molecular weight 2000, L/G=2m/ 4n, m=2000/44=45.5, then the composition and molecular weight of the copolymer can be obtained.

Molecular weight of copolymer=(weight ratio of 1+PDLLA/MPEG)×2000

In the formula: L is the integrated area of the composite peak at 5.1ppm, representing the methine group of polylactide;

G is the integrated area of the composite peak at 3.6ppm, representing the methylene group of polyoxyethylene;

m is the degree of polymerization of oxyethylene in the copolymer structural formula;

n is the degree of polymerization of D,L-lactide in the copolymer structure formula.

GPC was determined according to high-performance liquid chromatography (Chinese Pharmacopoeia 2020 Edition Four General Rules <0512>).

The chromatographic conditions adopt gel chromatographic column; differential refraction detection; tetrahydrofuran as mobile phase; flow rate 1.0mL/min, column temperature 40°C.

Determination method Take an appropriate amount of this product, add tetrahydrofuran to prepare a 1% solution, draw 20 μL, inject it into a liquid chromatograph, use polystyrene standard substance as a standard sample, and use GPC software to process the data.

(2) Detection method of related substances:

Chromatographic conditions:

Mobile Phase A: Water

Mobile Phase B: Acetonitrile

Gradient elution according to the table:

Diluent: acetonitrile-water-glacial acetic acid (100:100:0.1)

Chromatographic column: Octadecylsilane bonded silica gel as filler (4.6×250mm, 5μm)

Column temperature: 35°C;

Detection wavelength: 230nm;

Flow rate: 1.0mL/min;

Injection volume: 20 μL.

Test solution: Take 1 bottle of docetaxel polymer micelle freeze-dried preparation, add appropriate amount of diluent to dissolve and transfer to a 50ml measuring bottle, wash the vial with appropriate amount of diluent no less than 3 times and transfer all to the measuring bottle In the bottle, add diluent to dilute to the mark, shake well;

Control solution: Accurately measure 1ml of the test product, put it in a 100ml measuring bottle, add diluent to the mark, and shake well.

Excipient solution: Accurately weigh an appropriate amount of polyethylene glycol monomethyl ether-polylactic acid block copolymer (MPEG-PDLLA), add diluent to dissolve and dilute to make a solution containing about 9.6mg per 1ml.

Sensitivity solution: Take an appropriate amount of docetaxel reference substance, accurately weigh it, add diluent to dissolve and dilute to make a solution containing about 0.2 μg per 1 ml.

System suitability solution: Take an appropriate amount of reference substance for identification of docetaxel, weigh it accurately, add diluent to dissolve and dilute to make a solution containing about 0.4mg per 1ml.

System suitability requirements: In the system suitability solution chromatogram, the separation degree of docetaxel and impurities B, C and D should meet the requirements. In the sensitivity solution chromatogram, the signal-to-noise ratio of the docetaxel peak height should be greater than 10.

Analysis process: Determination according to high performance liquid chromatography (Chinese Pharmacopoeia 2020 Edition Sibu General Rules <0512>). Precisely measure the test solution and the reference solution, inject them into the liquid chromatograph respectively, and record the chromatograms.

Calculation formula:

Total impurity (%) = ∑% individual impurity

in:

A _SPL impurity peak area in the test solution

A The area of the main peak in the _STD control solution

f correction factor

Judgment criteria: if there are impurity peaks in the chromatogram of the test solution, deduct the excipient peaks, and calculate the impurity content by the main component self-comparison method multiplied by the correction factor (1.0). In the chromatogram of the test solution, peaks that are less than 0.05 times the area of the main peak of the control solution are ignored.

(3) Particle size and span detection method: take docetaxel polymer micelles freeze-dried preparation, add 20mL of 0.9% sodium chloride injection to prepare a solution, follow the guidelines for microparticle preparations (Chinese Pharmacopoeia 2020 Edition Four General Rules< 9014>) and particle size and particle size distribution determination method (Chinese Pharmacopoeia 2020 Edition Four General Rules <0982> third method) for determination.

Embodiment 1, the preparation of polyethylene glycol monomethyl ether-polylactic acid block copolymer

Add 4.50kg of polyethylene glycol monomethyl ether (2000) into the reaction kettle under the protection of nitrogen, fill with nitrogen, vacuumize, heat to 120°C, wait until it is completely melted, continue heating and keep the vacuum, and replace it with nitrogen after drying for 2 hours. Add 4.95kg of D,L-lactide under the protection of nitrogen, fill with nitrogen and evacuate to keep the vacuum in the reaction kettle; after stirring evenly, replace with nitrogen. Raise the temperature to 140°C, under the protection of nitrogen, add 18.000g of stannous isooctanoate, replace with nitrogen, and heat to 140±5°C under the protection of nitrogen to react for 5h. After the reaction, the product was cooled to 40°C, 36L of dichloromethane and 4.50kg of activated carbon were added, stirred for 1 hour to decolorize, filtered, and the filtrate was concentrated to dryness under reduced pressure.

Add 4.5L of dichloromethane, add 90L of cold anhydrous ether under stirring, let stand after stirring for 30 minutes, filter, and refine the filter cake twice according to the above operation process, and vacuum dry to obtain polyethylene glycol monomethyl ether-polylactic acid block Copolymer 6.204kg, the yield is 65.65%. The obtained copolymer was characterized by nuclear magnetic resonance, and the results are shown in FIG. 1 . The molecular weight is 3647, GPC analysis result: polydispersity coefficient PD=1.2.

Embodiment 2-4, preparation of polyethylene glycol monomethyl ether-polylactic acid block copolymer

The method is the same as in Example 1, except that the following materials are used to prepare polyethylene glycol monomethyl ether-polylactic acid block copolymer. The content of D,L-lactide in the prepared polyethylene glycol monomethyl ether-polylactic acid block copolymer was detected.

Embodiment 5, preparation of polyethylene glycol monomethyl ether-polylactic acid block copolymer

Add 20g of polyethylene glycol monomethyl ether (2000) into the reaction bottle under the protection of nitrogen, fill with nitrogen, vacuumize, keep the vacuum in the reaction bottle, heat to 120°C, wait until it is completely melted, continue heating and keep the vacuum, and dry for 2h Then replace with nitrogen. Add weighed 22g of D,L-lactide under the protection of nitrogen, fill with nitrogen and evacuate to keep vacuum in the reaction bottle; after stirring evenly, replace with nitrogen. The temperature was raised to 135° C., under nitrogen protection, 0.08 g of stannous isooctanoate was added, nitrogen replacement, nitrogen protection, and heated to 140° C. for 5 hours. After the reaction, the temperature of the product was lowered to 40° C., 160 ml of dichloromethane and 10 g of activated carbon were added, stirred for 1 h, and filtered. The filtrate was concentrated to dryness under reduced pressure. Use 20ml of dichloromethane to transfer the material in the rotary evaporator to the reaction flask, add 400ml of cold anhydrous diethyl ether under stirring, stir for 30min, let stand, and filter. The filter cake was refined twice according to the above operation process, and dried in vacuum to obtain 32 g of polyethylene glycol monomethyl ether-polylactic acid block copolymer. D,L-lactide content is 0.07%. GPC analysis results: polydispersity coefficient PD = 1.11. Inductively coupled plasma mass spectrometry (ICPMS) was used to detect the content of metal tin ions, and the result: the content of metal tin ions was 0.042ppm.

Embodiment 6, the preparation of polyethylene glycol monomethyl ether-polylactic acid block copolymer

Add 20g of polyethylene glycol monomethyl ether (2000) into the reaction bottle under the protection of nitrogen, fill with nitrogen, vacuumize, keep the vacuum in the reaction bottle, heat to 120°C, wait until it is completely melted, continue heating and keep the vacuum, and dry for 2h Then replace with nitrogen. Add weighed 22g of D,L-lactide under the protection of nitrogen, fill with nitrogen and evacuate to keep the vacuum in the reaction bottle; after stirring evenly, replace with nitrogen. The temperature was raised to 135°C, and under nitrogen protection, 0.08g of stannous isooctanoate was added, and heated to 140°C for 5 hours. After the reaction, the temperature of the product was lowered to 40° C., 160 ml of dichloromethane and 20 g of activated carbon were added, stirred for 1 h, and filtered. The filtrate was concentrated to dryness under reduced pressure. Use 20ml of dichloromethane to transfer the material in the rotary evaporator to the reaction flask, add 400ml of cold anhydrous diethyl ether under stirring, stir for 30min, let stand, and filter. The filter cake was refined twice according to the above operation process, and dried in vacuum to obtain 32 g of polyethylene glycol monomethyl ether-polylactic acid block copolymer. Inductively coupled plasma mass spectrometry ICPMS detects the content of metal tin ions, and the test result: the content of metal tin ions is 0.06ppm.

Embodiment 7, preparation of polyethylene glycol monomethyl ether-polylactic acid block copolymer

Add 150g of polyethylene glycol monomethyl ether (2000) into the reaction bottle under the protection of nitrogen, fill with nitrogen, vacuumize, keep the vacuum in the reaction bottle, heat to 120°C, wait until it is completely melted, continue heating and keep the vacuum, and dry for 2h Then replace with nitrogen. Add weighed 165g of D,L-lactide under the protection of nitrogen, fill with nitrogen, and vacuumize to keep the vacuum in the reaction product; after stirring evenly, replace with nitrogen. The temperature was raised to 135° C., under the condition of nitrogen protection, 0.6 g of stannous isooctanoate was added, replaced by nitrogen, under nitrogen protection, and heated to 140° C. for 5 hours. After the reaction, the temperature of the product was lowered to 40°C, 150ml of dichloromethane was added to dissolve it, and 3000ml of cold anhydrous ether was added under stirring, and after stirring for 30min, it was left to stand and filtered. The filter cake was refined twice according to the above operation process, and dried in vacuum to obtain 224 g of polyethylene glycol monomethyl ether-polylactic acid block copolymer. Inductively coupled plasma mass spectrometry ICPMS detects the content of metal tin ions, and the result: the content of metal tin ions is 350ppm.

Embodiment 8, the preparation of docetaxel polymer micelle lyophilized preparation

formula:

Preparation process: (1) Weigh the polyethylene glycol monomethyl ether-polylactic acid block copolymer prepared in Example 1 of the recipe amount, add it to 50% of the recipe amount of methanol and ultrasonically dissolve it until it is completely dissolved to obtain a methanol solution.

(2) Add the prescribed amount of citric acid into the above-mentioned methanol solution to dissolve. After the dissolution, add the prescribed amount of docetaxel and the remaining methanol, continue to dissolve, and then filter. Rotary steam for at least 1 h to obtain a docetaxel polymer gel film.

(3) Add 15kg of water for injection at a temperature of about 50-60°C to the round-bottom rotary steamer bottle filled with docetaxel polymer film, and place it in a rotary oscillator for hydration. After complete hydration, place the round-bottom flask Cool in cold water, docetaxel polymer micelles solution. Sterilizing and filtering, filling in vials and freeze-drying to obtain the freeze-dried preparation of docetaxel polymer micelles. Each bottle contains 20mg of docetaxel.

A small amount of the docetaxel polymer micelles freeze-dried preparation was taken and detected by transmission electron microscope after reconstitution. The results are shown in Figure 2, showing that the sample is approximately spherical and micelles have been formed.

Take a small amount of docetaxel polymer micelles freeze-dried preparation, detect the particle size and span, the particle size is 22nm, and the span is 0.73.

Embodiment 9, encapsulation efficiency detection experiment

The sample prepared in Example 8 was taken, dissolved in physiological saline to a concentration of 1 mg/ml, placed at room temperature (25±2° C.), and the encapsulation efficiency was detected at different time points.

Reference substance solution (containing about 0.2mg/ml docetaxel): Accurately weigh 10mg of docetaxel reference substance in a 50ml measuring bottle, add diluent to dissolve and dilute to the mark, shake well, and use it as a reference substance for content determination solution.

Test product stock solution (containing docetaxel 1mg/ml): take 5 bottles of this product (each bottle contains 20mg docetaxel), add 20ml of 0.9% sodium chloride injection respectively, vortex and mix for 10 minutes, As the test stock solution. Configure three copies in parallel.

Solution before centrifugation: Take 4ml of the stock solution of the test product, centrifuge at 10000G for 10 minutes, accurately measure 2.0ml of the supernatant, place it in a 10ml measuring bottle, dilute to the mark with diluent, shake well, and use it as the solution after centrifugation.

The HPLC chromatographic conditions are: ODS is used as filler, 0.043mol/L ammonium acetate aqueous solution-acetonitrile (45:55) is used as mobile phase, and the detection wavelength is 230nm.

The test results of the encapsulation efficiency at different times after reconstitution of the docetaxel polymer micellar lyophilized preparation are as follows:

The results show that the docetaxel polymer micelle freeze-dried preparation described in the present disclosure has excellent encapsulation efficiency, and the encapsulation efficiency is not less than 95% 24 hours after reconstitution.

Embodiment 10-12, the preparation of docetaxel polymer micelle lyophilized preparation

Docetaxel polymer micelles were prepared according to the following dosages, and related substances (total impurities) were investigated.

(1) Preparation of polyethylene glycol monomethyl ether-polylactic acid block copolymer

Add polyethylene glycol monomethyl ether (2000) into the reaction flask, fill with nitrogen, vacuumize, keep the vacuum in the reaction flask, heat to 100°C ~ 120°C, wait until it is completely melted, continue heating for 2h and keep the vacuum; nitrogen After replacement, add D,L-lactide, fill with nitrogen, and vacuumize to keep the vacuum in the reaction product; after nitrogen replacement, add 0.18% wt of stannous isooctanoate, nitrogen replacement, nitrogen protection , heated to 140°C and reacted for 5 hours. After the reactant was cooled to room temperature, the product was dissolved in dichloromethane equivalent to 0.5 times the weight of the reactant, and immediately added with cold anhydrous diethyl ether equivalent to 10 times the weight of the reactant for precipitation under stirring. , after stirring for 30min, let stand and filter. The filter cake was refined twice according to the above operation process, and the product was vacuum-dried to obtain a polyethylene glycol monomethyl ether-polylactic acid block copolymer.

(2) Docetaxel polymer micelles were prepared according to the preparation method in Example 8 and according to the dosage in the above table.

Take a small amount of the freeze-dried preparation of docetaxel polymer micelles prepared in Examples 10-12, and determine related substances (total impurities). The results are as follows

investigation Example 10 Example 11 Example 12 Related substances (total miscellaneous) 0.43% 0.28% 0.26%

As can be seen from the test results, MPEG and D, the L-lactide feed mass ratio is 1:1.1, 1:1.2 preparations are relative to MPEG and D, the L-lactide feed mass ratio is 1:1.0 preparation related substances (total Miscellaneous) content was significantly reduced.

Claims (27)

A kind of polyethylene glycol monomethyl ether-polylactic acid block copolymer, described polyethylene glycol monomethyl ether-polylactic acid block copolymer is D, L-lactide and polyethylene glycol monomethyl ether A block copolymer formed by polymerization, wherein the mass ratio of polyethylene glycol monomethyl ether to D, L-lactide is 1:1.05 to 1.25, and the polyethylene glycol monomethyl ether-polylactic acid block The segment copolymer is prepared by the following method: (a) adding polyethylene glycol monomethyl ether into the reactor, heating and melting under vacuum, and replacing with inert gas; (b) adding D,L-lactide, replacing with inert gas, Add a metal catalyst to ensure negative pressure or inert gas protection in the reactor, and then heat to 125-150°C for reaction; (c) cool down after the reaction, add the first organic solvent to dissolve, add the second organic solvent, filter, filter cake Vacuum drying to obtain polyethylene glycol monomethyl ether-polylactic acid block copolymer. The polyethylene glycol monomethyl ether-polylactic acid block copolymer according to claim 1, wherein the mass ratio of polyethylene glycol monomethyl ether to D, L-lactide is 1:1.1 to 1.2, Preferably 1:1.1. The polyethylene glycol monomethyl ether-polylactic acid block copolymer according to claim 1, wherein the molecular weight of the polyethylene glycol monomethyl ether in step (a) is 1000-20000, preferably 1000-5000, 1800-2200 is more preferable, and 2000 is still more preferable. The polyethylene glycol monomethyl ether-polylactic acid block copolymer according to claim 1, wherein the polyethylene glycol monomethyl ether in step (a) is heated to 60-130° C. under vacuum. Polyethylene glycol monomethyl ether-polylactic acid block copolymer according to claim 1, wherein, step (b) metal catalyst is stannous octoate. Polyethylene glycol monomethyl ether-polylactic acid block copolymer according to claim 5, wherein, the quality of described stannous isooctanoate accounts for D, L-lactide and polyethylene glycol monomethyl ether total mass 0.05 to 0.5 wt%, preferably 0.15 to 0.3 wt%. The polyethylene glycol monomethyl ether-polylactic acid block copolymer according to claim 1, wherein the reaction time of step (b) is 1-20 h, preferably 3-10 h, more preferably 4-6 h. Polyethylene glycol monomethyl ether-polylactic acid block copolymer according to claim 1, wherein, the first organic solvent described in step (c) is acetonitrile, acetone, dichloromethane, dimethylformamide, One or more of dimethyl sulfoxide, tetrahydrofuran, short-chain aliphatic alcohol or ethyl acetate, preferably one or more of dichloromethane, chloroform, methanol, ethanol, isopropanol, more preferably di Chloromethane; the consumption of the first organic solvent accounts for 0.1～100 times of volume (L/kg) of polyethylene glycol monomethyl ether quality, preferably 0.2～50 times of volume (L/kg), more preferably 0.5～20 times of volume (L/kg) /kg). Polyethylene glycol monomethyl ether-polylactic acid block copolymer according to claim 1, wherein, the second organic solvent described in step (c) is ether, isopropyl ether, methyl tert-butyl ether, normal One or more of heptane, cyclohexane, petroleum ether, preferably one or more of ether, isopropyl ether or methyl tert-butyl ether, more preferably ether; the amount of the second organic solvent accounts for The total mass of ethylene glycol monomethyl ether is 1-100 times the volume (L/kg), preferably 10-50 times the volume (L/kg), more preferably 15-20 times the volume (L/kg). Polyethylene glycol monomethyl ether-polylactic acid block copolymer according to claim 1, wherein, after step (b) reaction also comprises adding the 3rd organic solvent and gac, stir, filter, filtrate decompression concentrates step. The polyethylene glycol monomethyl ether-polylactic acid block copolymer according to claim 10, wherein the third organic solvent is dichloromethane, chloroform, ethyl acetate, isopropyl acetate, butylacetate One or more of esters, butanone, toluene, acetone, acetonitrile, dimethylformamide, dimethyl sulfoxide, tetrahydrofuran, 1,4-dioxane, short-chain fatty alcohols, preferably dichloromethane, One or more of chloroform, methyl alcohol, ethanol, isopropanol, more preferably dichloromethane; The consumption of the third organic solvent accounts for 0.1～100 times volume of polyethylene glycol monomethyl ether quality (L /kg), preferably 0.2 to 50 times the volume (L/kg), more preferably 0.5 to 20 times the volume (L/kg). Polyethylene glycol monomethyl ether-polylactic acid block copolymer according to claim 10, wherein, the consumption of described activated carbon accounts for D, L-lactide and polyethylene glycol monomethyl ether total mass 1～ 500wt%, preferably 5-200wt%, more preferably 5-75wt%, even more preferably 20-50wt%. The polyethylene glycol monomethyl ether-polylactic acid block copolymer according to claim 10, wherein, the metal tin ion content in the polyethylene glycol monomethyl ether-polylactic acid block copolymer is below 10ppm, preferably It is 1 ppm or less, more preferably 0.1 ppm or less. Polyethylene glycol monomethyl ether-polylactic acid block copolymer according to claim 1, is characterized in that, described polyethylene glycol monomethyl ether-polylactic acid block copolymer is prepared by the following method: ( a) Add polyethylene glycol monomethyl ether into the reactor, fill with nitrogen, vacuumize, heat and vacuum dry, and replace with inert gas; (b) add D,L-lactide, fill with nitrogen, vacuumize, and make the reactor Keep vacuum in the reactor, replace with inert gas, add metal catalyst under the protection of nitrogen, replace with nitrogen, ensure negative pressure or inert gas protection in the reactor, and then heat to 125-150°C for reaction; (c) Cool down after the reaction, add the first Dissolve the organic solvent, add a second organic solvent, filter, and vacuum-dry the filter cake to obtain polyethylene glycol monomethyl ether-polylactic acid block copolymer. A hydrophobic drug polymer micelle freeze-dried preparation, wherein the freeze-dried preparation comprises a hydrophobic drug and the polyethylene glycol monomethyl ether-polylactic acid block copolymer described in any one of claims 1 to 14 . The hydrophobic drug polymer micelle freeze-dried preparation according to claim 15, wherein the mass ratio of the hydrophobic drug to polyethylene glycol monomethyl ether-polylactic acid block copolymer is 0.01-0.15:1, Preferably 0.02-0.12:1, more preferably 0.03-0.10:1, still more preferably 0.04:0.96. The hydrophobic drug polymer micelle lyophilized formulation according to claim 15, wherein the lyophilized formulation further comprises a stabilizer. The hydrophobic drug polymer micelle freeze-dried preparation according to claim 17, wherein, the stabilizer is citric acid, hydrochloric acid, sorbic acid, lactic acid, tartaric acid, malic acid, phosphoric acid, acetic acid, adipic acid, One or more of malic acid, preferably one or more of citric acid, tartaric acid and fumaric acid, more preferably citric acid. The hydrophobic drug polymer micelle lyophilized preparation according to claim 17, wherein the mass ratio of the hydrophobic drug to the stabilizer is 1:0.00125-0.25, preferably 1:0.005-0.25, more preferably 1 :0.0125～0.125. The hydrophobic drug polymer micelle lyophilized preparation according to claim 15, wherein the hydrophobic drug is selected from taxane compounds, preferably, the hydrophobic drug is selected from docetaxel or paclitaxel, More preferably, the hydrophobic drug is selected from docetaxel. The hydrophobic drug polymer micelle lyophilized formulation according to claim 15, wherein the lyophilized formulation comprises the following components:

The preparation method of the hydrophobic drug polymer micelle freeze-dried preparation described in any one of claims 15 to 21, comprising the following steps: (1) adding the polyethylene glycol monomethyl ether described in any one of claims 1 to 14 - the polylactic acid block copolymer and the hydrophobic drug are dissolved in an organic solvent, and the organic solvent is removed by rotary evaporation to obtain a gel-like drug film, and then water is added to the drug film to dissolve and disperse the drug film to obtain a micellar solution; ( 2) The micellar solution prepared in step (1) is sterilized by filtration and freeze-dried to obtain a freeze-dried preparation of hydrophobic drug polymer micelles. The preparation method according to claim 22, wherein, step (1) is to mix the polyethylene glycol monomethyl ether-polylactic acid block copolymer, stabilizer and hydrophobic drug described in any one of claims 1 to 14 Dissolving in an organic solvent, removing the organic solvent by rotary evaporation to obtain a gel-like drug film, and then adding water to the drug film to dissolve and disperse the drug film to obtain a micellar solution. The preparation method according to claim 22 or 23, wherein, the organic solvent described in step (1) is selected from acetonitrile, acetone, methylene chloride, dimethylformamide, dimethyl sulfoxide, tetrahydrofuran, short-chain aliphatic Any one or several in alcohol, ethyl acetate, preferably any one or several in methyl alcohol, ethanol, isopropanol, acetonitrile, ethyl acetate, more preferably methyl alcohol; The organic described in step (1) The amount of solvent used is to add 0.5-12 ml of organic solvent per gram of polyethylene glycol monomethyl ether polylactic acid block copolymer, preferably 3-12 ml of organic solvent. The preparation method according to claim 22 or 23, wherein the amount of water added to the drug film in step (1) is 2 to 40 ml of water per gram of polyethylene glycol monomethyl ether polylactic acid block copolymer, preferably Ideally, add 10-20 ml of water per gram of polyethylene glycol monomethyl ether polylactic acid block copolymer, more preferably add 10-15 ml of water per gram of polyethylene glycol monomethyl ether polylactic acid block copolymer. The preparation method according to claim 22 or 23, wherein, in step (2), the conditions for removing the organic solvent by rotary evaporation are: rotation speed 10-150 rpm, temperature 20-80° C., and time 1-4 hours. Use of the hydrophobic drug polymer micelle freeze-dried preparation according to any one of claims 15 to 21 in the preparation of a drug for preventing or treating cancer.

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