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Rhamnose modified tumor whole-cell vaccine

A whole-cell vaccine, tumor cell technology, applied in the field of tumor whole-cell vaccine, can solve the problems of insufficient effect, complicated preparation process, no reports, etc., and achieves good application prospects, simple preparation method, and good stability.

Inactive Publication Date: 2014-04-09
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] In summary, there are still many problems in the preparation of tumor whole cell vaccines, and the preparation process is complicated, the cost is high, and the effect is not significant enough
However, the search found that there is no report on the whole-cell vaccine for tumors modified with rhamnose antigen, and there is no report on the method of modifying the whole-cell vaccine by chemical methods.

Method used

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  • Rhamnose modified tumor whole-cell vaccine
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  • Rhamnose modified tumor whole-cell vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1: chemical synthesis activates Rha

[0035] 1. Dissolve 10g Rha in 100ml dichloromethane, add Ac 2 O3 1.1mL, Et 3 N6 1.2mL, DMAP 0.67g, stirred overnight at 0°C. Then 20ml of methanol was added to terminate the reaction, and the crude product of tetraacetate of Rha was obtained after washing and drying.

[0036] 2. Take out 24g of Rha tetraacetate and dissolve in 150ml of anhydrous methanol, add 8.12ml of thiophenol, SnCl 2 5.92ml, placed at 0°C and stirred for 3h. After washing, drying and concentrating, it is purified by column chromatography to obtain the glucosinolate compound of Rha.

[0037] 3. Take 1 g of the above glucosinolate compound, add 0.39 g of 3-azidopropanol, 0.88 g of NIS, 2-3 μl of TMSOTf, and react at -30°C for 1.5 h. Add 2-3ml NaHCO 3 Stop the reaction. After washing, drying, concentration and purification by column chromatography, an intermediate with an azide end is obtained.

[0038] 4. Dissolve 0.52g of the above-mentioned az...

Embodiment 2

[0043] Example 2: Preparation of Rha-modified human melanoma M21 whole cell vaccine

[0044] Different concentrations of activated Rha were prepared and mixed with M21 cells to prepare the Rha-modified human melanoma M21 tumor whole cell vaccine. Among them, the preferred cell concentration is 4×10 6 / ml, the final concentration of activated Rha was 1mg / ml.

[0045] The reaction pH values ​​were set to 7.4 and 8.5 respectively. After reacting at room temperature for 1 h, they were washed with PBS, and then FITC-labeled anti-rhamnose antibody was added. After washing, the efficiency of coupling Rha to M21 cells was detected by flow cytometry.

[0046] The results of flow cytometry showed that Rha was successfully coupled to the surface of M21 cells, ( figure 2 ) and when the reaction pH was 8.5, the proportion of M21 cells coupled with Rha reached over 92%, while when the pH was 7.4, the proportion of M21 cells coupled with Rha was only 75.92%.

[0047] The Rha-modified M21...

Embodiment 3

[0049] Example 3: Preparation of Rha-modified mouse melanoma B16 whole-cell vaccine

[0050] Different concentrations of activated Rha were prepared and mixed with B16 cells to prepare the Rha-modified mouse melanoma B16 tumor whole-cell vaccine. Among them, the preferred cell concentration is 4×10 6 / ml, the final concentration of activated Rha is 0.5mg / ml, 1mg / ml, 2mg / ml, 4mg / ml respectively, the reaction pH value is 8.5, after 1h reaction at room temperature, wash with PBS, then add FITC-labeled anti-rhamnose antibody , and then the efficiency of B16 cells coupled to Rha was detected by flow cytometry after washing.

[0051] The results of flow cytometry showed that Rha was successfully coupled to the surface of B16 cells, ( image 3 ) and when the activated Rha concentration was above 1 mg / ml, the proportion of B16 cells coupled with Rha reached over 98%.

[0052] The Rha-modified B16 tumor cells prepared above were irradiated with a total dose of 40Gy of X-rays to inac...

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Abstract

The invention discloses a rhamnose modified tumor whole-cell vaccine consisting of complete tumor cells, wherein the complete tumor cells have rhamnose antigens on surfaces through a chemical method and lose differentiation and proliferation capacities through inactivation treatment. The tumor vaccine disclosed by the invention is simple in preparation, good in stability and low in cost, can overcome the defects of poor effect, easy relapse and great side effect of an existing tumor treatment method, is safer and more effective to treat tumors, is theoretically suitable for treatment of various malignant tumors and cancers of human, and has very broad application prospects.

Description

technical field [0001] The invention relates to a cell-based vaccine, in particular to a rhamnose (Rha)-modified tumor whole-cell vaccine. Background technique [0002] Malignant tumors are one of the major diseases that threaten human health and life. Due to the poor therapeutic effects and side effects of traditional surgery, radiotherapy, and chemotherapy, immunotherapy, a new treatment model, has gradually been placed with high hopes, and tumor vaccines have emerged as the times require. [0003] Currently, tumor vaccines are mainly therapeutic vaccines. Therapeutic tumor vaccines can induce systemic specific anti-tumor effects through active immunization. Compared with surgical treatment and radiotherapy, they have a wider range of effects and are especially suitable for multiple focal tumors, extensively metastatic tumors or non-solid tumors (such as leukemia); the body's own strength can be mobilized to achieve anti-tumor effects, and it has fewer adverse reactions ...

Claims

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Application Information

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IPC IPC(8): A61K39/00A61P35/00
Inventor 王鹏张化杰商文静吴宝林刘军陈敏马中瑞
Owner SHANDONG UNIV
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