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Liposome preparation for tumor therapeutic vaccine and preparation method thereof

A technology of liposome preparation and tumor treatment, which is applied in the direction of liposome delivery, anti-tumor drugs, medical preparations of non-active ingredients, etc., and can solve the problem of inability to produce therapeutic vaccine protein antigen cross-presentation and inability to effectively protect Antigen etc.

Inactive Publication Date: 2010-09-01
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Existing methods do not effectively protect antigens from enzymatic degradation in endosomes or lysosomes, and cannot produce the protein-antigen cross presentation necessary for therapeutic vaccines

Method used

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  • Liposome preparation for tumor therapeutic vaccine and preparation method thereof
  • Liposome preparation for tumor therapeutic vaccine and preparation method thereof
  • Liposome preparation for tumor therapeutic vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Dissolve 20 mg / mL chicken ovalbumin (ovalbumin) solution with 0.3% sodium carbonate solution, adjust isotonicity with sodium chloride, adopt a lipid prescription with a molar ratio of lecithin (EPC) / cholesterol (cholesterol) 1 / 1, pass through ethanol Prepare liposomes by injection method, dissolve 20mg EPC and 10mg cholesterol in 1mL ethanol, filter the ethanol solution with a microporous membrane, and set aside. Add 14mL of 20mg / mL ovalbumin in 0.3% sodium carbonate solution into a 50mL flask containing a stirring bar, and then rapidly inject 1mL of lipid ethanol solution into the liposome-forming suspension under high-speed stirring. Ethanol was removed by dialysis, and then the prepared liposomes were ultrasonicated so that the average particle size was 500 nanometers.

[0028] Separation of liposome and free protein by using a molecular sieve gel column, 10% Triton solution to disrupt the liposome membrane, and quantify the concentration of ovalbumin in it by HPLC, ...

Embodiment 2

[0034]Prepare liposomes by freeze-drying, dissolve ovalbumin with 0.9% sodium bicarbonate to form a 2 mg / mL protein solution, adjust isotonicity with sodium chloride, dissolve 40 mg DPPC and 10 mg cholesterol in chloroform solution, and place in an eggplant-shaped bottle In the process, chloroform was removed under low pressure on a rotary evaporator, 1 mL of ovalbumin solution was added to hydrate, liquid nitrogen was frozen, freeze-dried overnight, and rehydrated to obtain a liposome suspension. The prepared liposomes were repeatedly frozen and thawed and ultrasonicated, and then passed through 1000nm, 400nm, 200nm, 100nm membranes successively to make the average particle size about 100nm.

[0035] The liposome and free protein were separated by molecular sieve gel column, the liposome membrane was broken with 10% Triton solution, and the concentration of ovalbumin was quantified by HPLC to determine the components of the prepared liposome. The sodium bicarbonate concentrat...

Embodiment 3

[0041] Prepare liposomes by film dispersion method, dissolve 5mg DPPC, 0.6mg cholesterol and 0.4mg DOTAP in chloroform solution, put them into an eggplant-shaped bottle, put them in a water bath at 30°C, remove the chloroform by rotary evaporation to form a film, and keep vacuum for 1 hour To remove the solvent, dissolve the ovalbumin protein in 3% sodium bicarbonate to a 10mg / mL solution, add 1mL ovalbumin solution to the eggplant-shaped bottle, ultrasonically oscillate, and place at room temperature for 2h to form a translucent lipoplex suspension . The obtained liposomes are subjected to high-pressure milk homogenization, so that the average particle size thereof is about 200 nm.

[0042] Utilize ultracentrifugation to separate liposome and free protein, 10% Triton solution breaks liposome membrane, and HPLC quantifies ovalbumin concentration therein, so as to measure the component of prepared liposome. The sodium bicarbonate concentration in the final liposome is 3%, the ...

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Abstract

The invention relates to a preparation and a preparation method thereof, which belong to the technical field of anti-tumor medicament. The preparation comprises the following components in percentage by weight: 0.3 to 3 percent of hydrocarbonate or carbonate, 0.0001 to 5 percent of tumor proteantigen, 0.1 to 10 percent of lipid and the balance of water. The preparation can promote the cross-presentation of protein or polypeptide antigen loaded by the preparation so as to cause specific cellular immunity and fulfill the purpose of curing tumor or virus infection.

Description

technical field [0001] The invention relates to a preparation in the technical field of anti-tumor drugs and a preparation method thereof, in particular to a liposome preparation for tumor therapeutic vaccines and a preparation method thereof. Background technique [0002] Human beings are constantly fighting against diseases in the process of their own development. With the development of modern medicine, many major diseases that threaten human life have been controlled. In particular, the discovery of penicillin has brought some breakthroughs in antibiotic research, which has brought many fatal infectious diseases under control in the past, and the average life expectancy of human beings has been improved. Great extension. However, there are still many critical diseases that cannot be effectively treated, and tumors are the most typical drugs among them. According to the data in 2000, there are 10 million new cancer cases every year in the world, and there is an upward t...

Claims

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Application Information

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IPC IPC(8): A61K9/127A61K39/00A61K47/02A61K47/34A61K47/28A61K47/24A61P35/00
Inventor 徐宇虹陈剑
Owner SHANGHAI JIAO TONG UNIV
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