Novel tumor vaccine preparation method

A tumor vaccine and a new technology, applied in the field of anti-tumor, can solve the problems of low vaccine activation efficiency, limited large-scale use, poor clinical efficacy, etc., and achieve easy large-scale production, good biosafety and biocompatibility, Addressing the effect of low immunogenicity

Pending Publication Date: 2019-08-16
迪亚思生命科技(武汉)有限公司
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0003] At present, clinically approved tumor vaccines include dendritic vaccine (DC vaccine), cervical cancer vaccine and individualized sipuleucel-T vaccine, but these vaccines have low activation efficiency, few antigens, poor clinical efficacy and high price. limits its large-scale clinical use

Method used

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Embodiment 1

[0029] The invention provides a preparation method of a novel tumor vaccine, which is prepared by mixing tumor cell lysates treated with hypochlorous acid (HClO) and an immune activator, and the immune activator is specifically CpG.

[0030] The preparation method of the tumor cell lysate after the hypochlorous acid (HClO) treatment, the specific steps are as follows:

[0031] 1): Primary tumor cells are cultured after fresh tumor tissue is obtained, and primary tumor cells are obtained, and the tumor tissue is an autologous isolated tumor tissue;

[0032] 2): Treat primary tumor cells with hypochlorous acid (HClO). Before treatment with hypochlorous acid (HClO), the tumor cells need to be washed with HBSS balanced salt solution for 2-3 times. The concentration of hypochlorous acid (HClO) is 60-80μM, the supernatant was collected 2 hours after the treatment with hypochlorous acid (HClO), and the supernatant obtained after the cells were treated with hypochlorous acid (HClO) wa...

Embodiment 2

[0039] refer to Figure 1-2 , assessing the activation of DC cells by novel tumors:

[0040] Bone marrow stem cells were obtained from the femur and tibia of male C57 mice aged 6-8 weeks, cultured in 1640 medium containing M-CSF for 5 days to obtain immature DC cells, and then planted in 24-well plates, divided into PBS group, Cell lysate group group (obtained by repeated freezing and thawing), HClO-lysate group, each group took 40 μl of the corresponding solution and incubated with immature DC cells to stimulate the maturation of DC cells. After 24 hours, CD11c, CD80, and CD86 cells were detected by flow cytometry Activation (such as figure 1 shown), the experimental results showed that having a novel tumor vaccine can promote the maturation of DC cells.

Embodiment 3

[0041] Example 3: Research on anti-tumor effect of novel tumor vaccine preventive therapy

[0042] The C57 mice were randomly divided into 3 groups, the first group of mice were injected with 50 μl PBS at the tail base + foot pads, the second group were injected with 50 μl Cell lysate at the tail base + foot pads, the third group were injected with 50 μl HClO-lysate at the tail base + foot pads, At this time, it was recorded as the first day, and the drug was given once every other day, for a total of 3 times. On the seventh day, malignant melanoma cells B16 (2*10 5 / 100μl / only), the survival situation was monitored regularly, experiments showed that the new tumor vaccine can significantly inhibit tumor growth.

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Abstract

The invention relates to the antitumor technical field, in particular to a novel tumor vaccine preparation method. Tumor cell lysis buffer subjected to treatment with hypochlorous acid is mixed with an immune activation agent to form a novel tumor vaccine. The method includes steps: 1) after fresh tumor tissues are obtained, performing primary tumor cell culture to obtain primary tumor cells; 2) subjecting the primary tumor cells obtained in the step 1) and culture solution to treatment with hypochlorous acid, collecting mixed solution obtained after cell treatment with hypochlorous acid to obtain required lysis buffer and apoptotic tumor cell debris mixture; 3) centrifuging the mixed solution obtained in the step 2), and repeatedly concentrating and purifying to obtain concentrated solution containing cell lysis buffer; 4) performing radiation inactivation on the cell lysis buffer purified and concentrated in the step 3). Problems of low immunogenicity, inaccuracy and low immunoreaction activation efficiency of common tumor antigen vaccines can be solved.

Description

technical field [0001] The invention relates to the field of anti-tumor technology, in particular to a preparation method of a novel tumor vaccine. Background technique [0002] With the rapid development of tumor genomics and immunology, immunotherapy has become the fourth treatment after radiotherapy, chemotherapy and surgery. The targeting of radiotherapy and chemotherapy is poor, and it is easy to damage normal cells and cause adverse reactions. Immunotherapy can specifically recognize antigens on the surface of tumor cells, target and kill tumor cells while reducing the occurrence of side effects. In immunotherapy, tumor vaccines are one of the research hotspots in recent years. Tumor vaccines express specific and immunogenic tumor antigens, such as tumor cell lysates, tumor-associated proteins or antigenic polypeptides, and related genes expressing tumor antigens, etc., with the assistance of adjuvants such as cytokines and chemokines Activate the body's immune syst...

Claims

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Application Information

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IPC IPC(8): A61K39/00A61K39/39A61P35/00
CPCA61K39/0011A61K39/39A61P35/00A61K2039/55561A61K2039/5152
Inventor 金红林黄浩聂晓芬周洁洪磊潘雄华
Owner 迪亚思生命科技(武汉)有限公司
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