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Method for preparing lipidosome coated with protein drugs

A technology of liposomes and lipid substances, applied in the directions of liposome delivery, peptide/protein components, pharmaceutical formulations, etc., can solve the problems of waste of raw materials, inability to reflect the advantages of liposome preparations, and inability to achieve effective doses, etc. , to achieve the effect of increasing the application value

Active Publication Date: 2014-10-08
SHENYANG PHARMA UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Low encapsulation efficiency will not only cause waste of raw materials, but also may not reach the effective dose, which cannot reflect the advantages of liposome preparations

Method used

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  • Method for preparing lipidosome coated with protein drugs
  • Method for preparing lipidosome coated with protein drugs
  • Method for preparing lipidosome coated with protein drugs

Examples

Experimental program
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Effect test

Embodiment 1

[0031] This embodiment is to adopt the method proposed by the present invention to prepare bovine serum albumin nanoliposomes. Step a: Preparation of microemulsion phase. Weigh 12mg of phosphatidylcholine, add 5ml of ether, and evaporate under reduced pressure to form a film. Add 5ml of diethyl ether to redissolve the membrane, add 8mg of Cremophor EL, and add dropwise 0.2ml of bovine serum albumin aqueous solution with a concentration of 2mg / ml under magnetic stirring until light blue opalescence, and the microemulsion phase is obtained. The ratio of oil phase, water phase and surfactant is optimized by pseudo ternary phase diagram: using Cremophor EL and phospholipid as mixed emulsifier according to Km is 9:1, 8:2, 7:3, 6:4, 5: 5. Mix 4:6, 3:7, 2:8, 1:9 evenly, ether is the oil phase, add distilled water drop by drop under magnetic stirring, observe the phenomenon of the system from turbid to clear or from clear to turbid, and record the critical The percentages of each co...

Embodiment 2

[0035] Step a: Preparation of microemulsion phase. Weigh 24 mg of phosphatidylcholine, add dichloromethane, and evaporate under reduced pressure to form a film. Add 9ml of diethyl ether to redissolve the membrane, add 16mg of Cremophor EL, add dropwise 0.6ml of 1mg / ml bovine serum albumin aqueous solution under magnetic stirring until light blue opalescence, and obtain the microemulsion phase.

[0036] Step b: Preparation of micellar phase. Weigh 30 mg of phosphatidylcholine, 5 mg of cholesterol, and 5 mg of cholesterol succinate, add an appropriate amount of dichloromethane, and evaporate under reduced pressure to form a film. Add 4ml of distilled water and 0.5m of propylene glycol to hydrate at 50°C to obtain the micellar phase;

[0037] Step c: the micellar phase was added to the microemulsion phase, and the probe was sonicated in an ice bath for 10 minutes, then the ether was removed by rotary evaporation at 30° C., and the remaining ether in the system was further remov...

Embodiment 3

[0039] Preparation of ovalbumin nanoliposomes. Step a: Preparation of microemulsion phase. Weigh 14 mg of phosphatidylcholine, add 5 ml of dichloromethane, and evaporate under reduced pressure to form a film. Add 5ml of diethyl ether to redissolve the membrane, add 406mg of Cremophor RH, and add dropwise 0.5ml of ovalbumin phosphate buffer with a concentration of 3mg / ml under magnetic stirring until light blue opalescence is obtained to obtain the microemulsion phase.

[0040] Step b: Preparation of micellar phase. Weigh 42 mg of distearoylphosphatidylethanolamine, 5 mg of positively charged lipid carbamoyl cholesterol, and 5 mg of cholesterol succinate, add appropriate amount of dichloromethane, and evaporate under reduced pressure to form a film. Add 2ml of phosphate buffer and 1ml of ethanol to hydrate at 60°C to obtain the micellar phase;

[0041] Step c: According to step c in Example 1, the average particle size of ovalbumin nanoliposomes was determined to be 68.68nm,...

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Abstract

The invention relates to a novel method for preparing small-particle-size high-encapsulation-efficiency lipidosome coated with protein drugs and belongs to the technical field of biological medicines. The preparation process comprises the following steps: a, dissolving lipids for forming the lipidosome in an organic solvent, mixing the lipids, a protein solution and a proper amount of surfactant according to a proper ratio, and preparing a micro-emulsion phase; b, dissolving the lipids for forming the lipidosome in an organic solvent, mixing and hydrating by using water or a buffer solution or alcohol according to a proper ratio, and preparing a micellar phase; and c, mixing the micro-emulsion phase and the micellar phase, reversing the phase b into a reverse micelle, reversing the phase b again in the process of removing the organic solvent under reduced pressure, finally covering the surface of the phase a micelle with the reverse micelle, and forming the lipidosome. The encapsulated protein drug lipidosome has the average particle size of about 80nm, and the encapsulation ratio can be over 90 percent. The preparation process is simple in process, the production cost is reduced, and industrial production is easily realized.

Description

technical field [0001] The invention relates to the field of biopharmaceuticals, in particular to a method for preparing liposomes with small particle diameters and high encapsulation efficiency of novel protein drugs. Background technique [0002] Nanoliposomes (nanoliposomes) refer to liposomes with a particle size of less than 100nm. The structure is mostly single-chamber liposomes. Hydrophobic and amphiphilic drugs are directly delivered to the target tissue to exert their pharmacological effect, thereby improving the therapeutic index of the drug, reducing the therapeutic dose of the drug and reducing the toxicity of the drug. [0003] At present, liposome preparation methods include: reverse evaporation method, thin film dispersion method, ethanol injection method, active drug loading method, etc., but these methods have low encapsulation efficiency of protein drugs, uncontrollable particle size, and complicated operation. Waiting is not enough. For example: the reve...

Claims

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Application Information

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IPC IPC(8): A61K9/127A61K47/24A61K47/28A61K48/00A61K38/38A61K38/08
Inventor 李可欣常莎莎陈大为王中彦
Owner SHENYANG PHARMA UNIVERSITY
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