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Process for preparing xenomal or autogenous tumor vaccine of liposome

A technology of tumor vaccines and liposomes, which is applied in the field of preparation of biological products, can solve the problems of lack of tumor inhibition rate, weak effect of enhancing antigenicity, weakened antigenicity, etc., and achieves advanced preparation methods, high purity and content, and effective lasting effect

Inactive Publication Date: 2004-03-17
广州市肿瘤医院 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The autovaccine method of this preparation has the following deficiencies: (1) the autologous vaccine material source is limited and may contain the composition that suppresses immune response; The antigenic components are destroyed, and the antigen is blocked by aldehydes, which weakens the antigenicity. The vaccine prepared by this method has not been tested for the antigenic protein, and its amount is difficult to estimate; (3) the added BCG or PPD acts as an adjuvant for a short time , the effect of enhancing antigenicity is weak; (4) The results of animal experiments show that the tumor weights of the treatment group and the control group are similar, only the number of metastatic tumors and the survival days of animals are different, without statistical analysis, it is difficult to accurately evaluate its tumor inhibition. Effect
Its disadvantages are: (1) the above-mentioned autologous vaccine materials are limited and can only be used for autologous; (2) the functions of the components of the Chinese patent medicine used are not clear; (3) the intact tumor cells are treated with aldehydes to block the antigen, and the antigenicity is weakened ; (4) Animal experiments lack the tumor inhibition rate, and it is not enough to evaluate the tumor inhibition effect only by the survival number of animals

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: Fresh specimens of poorly differentiated adenocarcinoma of the human mammary gland were prepared as liposome-encapsulated tumor vaccines for treating animal tumors. (Example of therapeutic application of liposome-encapsulated heterogeneous tumor vaccine)

[0030] (1) Weigh the fresh specimens of human breast cancer surgically removed under sterile conditions, strip off the surface fat and other non-tumor tissues, and cut into about 1mm 2 sized pieces.

[0031] (2) Put the obtained tumor fragments into a tissue grinder, grind 1 g of tissue with 4 ml of normal saline to form a tissue homogenate, and then filter the tumor cell suspension through a 200-mesh sieve.

[0032] (3) The obtained tumor cell suspension is observed by the trypan blue exclusion staining method, and the number of viable tumor cells is above 95%;

[0033] (4) The obtained tumor cell suspension was washed 3 times with Hank's solution, and then repeatedly frozen and thawed 3 times in liquid ...

Embodiment 2

[0042] Example 2: A liposome-encapsulated autologous tumor vaccine prepared from tumor-associated antigen extracted from a mouse breast cancer cell line (Scc891) was used to treat animal tumors.

[0043] The experimental steps are as follows:

[0044] (1) Take the cell suspension of mouse spontaneous breast cancer cell line (Scc891), the cell content is 1×10 7 / ml.

[0045] (2) Take the tumor cell suspension and use the trypan blue exclusion method to observe that the number of viable tumor cells is above 95%;

[0046] (3) repeated freezing and thawing of the obtained tumor cell suspension to inactivate the tumor cells;

[0047] (4) Resuspend with 2.5% (V / V) pH 7.4 n-butanol PBS solution, shake gently at room temperature for 20 minutes;

[0048] (5) The above-mentioned tumor cell suspension was centrifuged at 500×g at 4°C for 10 minutes, and the supernatant was then centrifuged at 2000×g at 4°C for 20 minutes, and the supernatant was taken.

[0049] (6) The obtained supern...

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Abstract

A process for preparing xenogenic or autologous tumor vaccine of liposome includes such steps as taking the fresh specimen of human tumor, preparing suspension of tumor cells; freeze-thaw step to break off and deactivate tumor cells; n-butanol suspending to remove lipid; centrifugal step to remove karyon; lowry's test to test qualification of supernatant; and coating with liposome as carrier and wrapping with adjuvant to make vaccine. Its advantages include advanced and efficient extraction process, no toxic by-effect, and durable and slow release.

Description

technical field [0001] The invention relates to a preparation method of a biological product, namely a preparation method of a tumor antigen vaccine. Background technique [0002] The application of tumor vaccines to induce specific active immune anti-tumor responses in tumor hosts is one of the effective methods for tumor biotherapy. It has received extensive attention from scholars at home and abroad. We have carried out research on autologous tumor vaccines and have made some progress (the paper: "Experimental Research on Vaccine Active Immunotherapy" was published in "Chinese Journal of Zoonoses" 1996; 12(3): 30). Based on the success of animal experiments, we have carried out research on the clinical application of autologous tumor vaccines. We have studied more than 300 cases of various tumors such as malignant melanoma, leiomyosarcoma, lung cancer, esophageal cancer, and liver cancer. The vaccine has the characteristics of safety, no toxic and side effects, and obvi...

Claims

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Application Information

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IPC IPC(8): A61K9/127A61K39/00A61K45/00A61P35/00
Inventor 甘岫云
Owner 广州市肿瘤医院
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