Combined medicine containing berberine hydrochloride and PD1-antibody for preventing and/or treating tumor

A technology of berberine hydrochloride and PD-1, which is applied in the field of biomedicine to achieve the effects of improving sensitivity, maintaining stable body weight, and reducing side effects of treatment

Inactive Publication Date: 2019-01-18
广东美赛尔细胞生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no literature report on the combination of berberine hydrochloride and PD-1 antibody drugs to treat tumors and reduce the side effects of treatment

Method used

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  • Combined medicine containing berberine hydrochloride and PD1-antibody for preventing and/or treating tumor
  • Combined medicine containing berberine hydrochloride and PD1-antibody for preventing and/or treating tumor
  • Combined medicine containing berberine hydrochloride and PD1-antibody for preventing and/or treating tumor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1, the preparation of berberine hydrochloride

[0030] Take 100 g of Coptis rhizome powder, extract twice with 900 ml of 50% ethanol under reflux, 1 hour each time, combine the two extracts, concentrate under reduced pressure until the extract is brownish-red syrupy. Add 1% acetic acid to the syrupy Coptis chinensis extract to dissolve it, suction filter it while it is hot, add concentrated hydrochloric acid dropwise to the filtrate to adjust the pH to 1.0, leave it to cool at room temperature, and yellow granular substances will precipitate out, filter it with suction, and use Wash twice with ice-cold water, once with acetone, and dry at 50°C to obtain crude berberine hydrochloride. The crude product was dissolved in water (70ml / 1g crude product) and heated, and concentrated hydrochloric acid was added dropwise to adjust the pH value to 2-3, then filtered while it was hot, and allowed to cool slowly at room temperature. After precipitation of crystals, suct...

Embodiment 2

[0031] Embodiment 2, preparation of mouse Nivolumab antibody for test

[0032] The variable region of the expression PD-1 antibody 5C4 gene was fused with the constant region of the mouse IgG2B gene to form a hybrid antibody. The gene was synthesized and cloned into pEGFP-C1 plasmid, and PD-1 was expressed by transient transfection in 293F cells. After being purified by Protein A column, molecular sieve and endotoxin removal column, the purity reaches 98%, and the endotoxin is lower than 1 EU / ml, dissolved in phosphate buffer, freeze-dried and stored.

experiment example 1

[0033] Experimental Example 1: Inhibitory effect of berberine hydrochloride combined with Nivolumab on the growth of subcutaneous transplanted tumors in animals and its toxic effect on animals.

[0034] 1.1 Test drugs: berberine hydrochloride prepared in Example 1 and Nivolumab prepared in Example 2.

[0035] 1.2 Experimental animals: 32 C57BL / 6 mice, female, weighing about 20 g, 6 weeks old, SPF grade, all mice had free access to food and water, and were raised at room temperature (25±2)°C. Feed and water are all autoclaved, and the entire feeding process is SPF grade.

[0036] 1.3 Experimental method:

[0037] 1.3.1 Establishment and intervention of tumor-bearing mouse model

[0038]Four subcutaneous xenograft tumor models were prepared respectively: liver cancer cell line Huh7, lung cancer cell line A549, breast cancer cell line MCF-7, and colon cancer cell line Lovo. 6 / ml of cell suspension, C57BL / 6 mice were injected with 200ul of cell suspension under the armpit of t...

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PUM

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Abstract

The invention belongs to the field of biomedicine, and specifically relates to a combined medicine containing berberine hydrochloride and PD1-antibody for preventing and / or treating tumor. The medicine comprises preparations of different specifications of units of berberine hydrochloride and PD1- Antibody, said PD- 1 antibody is preferably Nivolumab. The invention accidentally finds that the combination of berberine hydrochloride and Nivolumab improves the sensitivity of the experimental tumor-bearing mice to Nivolumab drugs, and the weight of the experimental tumor-bearing mice is kept stableafter the treatment course is finished; At the same time, the combination of the two has a synergistic effect, and the inhibiting ability of the combination is significantly superior to that of usingNivolumab alone, and the combination has made remarkable progress compared with the prior art.

Description

technical field [0001] The invention belongs to the field of biomedicine, and specifically relates to a combined medicine for preventing and / or treating tumors containing berberine hydrochloride and PD1-antibody. Background technique [0002] Tumor immunotherapy has become the fourth largest tumor treatment after surgery, radiotherapy and chemotherapy, and it is also the most popular tumor treatment at present. At present, tumor immunotherapy mainly includes immune vaccines, immune checkpoint inhibitor therapy, adoptive immune cell therapy and cytokine therapy, etc. Among them, immune checkpoint inhibitor therapy has attracted much attention due to its remarkable clinical efficacy, and "PD- 1 / PD-L1" has shown encouraging clinical effects in the field of cancer research and treatment, bringing new hope for the rebirth of patients. [0003] "PD-1 / PD-L1" is a drug target on T cells, an important part of the human immune system. Drugs designed for this target can activate the i...

Claims

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Application Information

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IPC IPC(8): A61K39/395A61K31/4375A61P35/00
CPCA61K31/4375A61K39/3955A61P35/00A61K2300/00
Inventor 王瑶梅建勋
Owner 广东美赛尔细胞生物科技有限公司
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