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Application of cyclic dinucleotide cGAMP combined PD1 antibody (Nivolumab) in preparation of antitumor drugs

A cyclic dinucleotide and anti-tumor technology, applied in the field of biomedicine

Inactive Publication Date: 2017-05-24
LIAOCHENG CITY ORIENT BIOMEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, there is no literature report on the combined use of cyclic dinucleotide cGAMP and PD1-PDL1 interaction inhibitory antibody in the treatment of tumors

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0009] Example 1 ： cGAMP preparation of

[0010] cGAMP (cyclized-GMP-AMP) is catalyzed by cyclized cGMP-AMP dinucleotide synthetase (cGAS) under the conditions of activation after binding to DNA according to the literature method. The purity is above 98%. (Pingwei Li, et al., Immunity, 2013, 39(6), 1019-1031.)

Embodiment 2

[0011] Example 2 ： Nivolumab preparation of

[0012] The variable region of Nivolumab 5C4 was fused with the constant region of mouse IgG2b to form a hybrid antibody. The gene was synthesized by Nanjing GenScript Biotechnology Co., Ltd. and cloned into the pTT3 plasmid. Plasmid extraction kits, Protein A chromatography columns, molecular sieves, endotoxin removal columns, and endotoxin determination kits were purchased from General Medical or Nanjing GenScript Biotechnology Co., Ltd.

[0013] Transient expression of Nivolumab 5C4 in 293F cells. After purification by Protein A column, molecular sieve and endotoxin removal column (purchased from General Medical), the purity is 98%, and the endotoxin is less than 1 EU / ml. Proteins were dissolved in phosphate buffered saline, freeze-dried and stored.

Embodiment 3

[0014] Example 3 ： Anti-tumor animal experiment of cGAMP combined with Nivolumab

[0015] The tumor-bearing mouse model was used to detect the inhibitory effect of cGAMP and Nivolumab combined with antineoplastic drugs on the growth of subcutaneous transplanted tumors in animals and the toxicity to animals.

[0016] Test drug

[0017] Name: cGAMP, Nivolumab

[0018] Appearance: white powder

[0019] Vehicle: normal saline.

[0020] Preparation method: Prepare the solution with the required concentration with physiological saline solution before use.

[0021] Test drug concentration: cGAMP: 5mg / ml, 10mg / ml, 20mg / ml; Nivolumab, 10mg / ml.

[0022] experimental animals

[0023] Species, strains, gender, body weight, source, certificate of conformity

[0024] C57BL / 6 mice, female, weighing 20 g, 8 weeks old, SPF grade, were purchased from Shanghai Slack Experimental Animal Co., Ltd. [Experimental animal quality certificate number: SCXK (Shanghai) 2007-0005].

[0025] ...

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PUM

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Abstract

The invention relates to the technical field of biological medicines, and relates to application of cyclic dinucleotide cGAMP combined PD1 antibody (Nivolumab) in preparation of antitumor combined drugs. Studies indicate that the cGAMP combined Nivolumab can significantly inhibit the growth of a variety of tumor cells, has obvious anti-tumor effect, and can be used for preparing the antitumor combined drugs. A C57BL / 6 mouse subcutaneous transplantation tumor model shows that the cGAMP combined Nivolumab has obvious inhibition effect on adenocarcinoma cell line MC38, lung cancer cell line LLC, breast cancer cell line EO771, melanoma cell line B16-F10 and colon cancer cells MC38, and the obvious inhibition effect is superior to single drug use effect, so that the cGAMP can be combined with the Nivolumab for the treatment of tumors.

Description

Technical field [0001] The invention belongs to the field of biomedicine technology, and specifically relates to the application of a cyclic dinucleotide (cGAMP) combined with Nivolumab in anti-tumor and in the preparation of anti-tumor drugs. Background technique [0002] Tumor is currently a major disease that seriously endangers human life and health. Its specific manifestations are excessive cell proliferation and abnormal differentiation, which are difficult to suppress. According to WHO expert predictions, by 2020, the number of cancer incidences in the global population will reach 20 million, and the number of deaths will reach 12 million. Therefore, tumors will become the largest killer of mankind in this century and pose the most serious threat to human survival. . In China, the incidence and mortality rates of lung cancer, colorectal cancer, gastric cancer, liver cancer, etc. are at the forefront of various malignant tumors. According to statistics from the Nation...

Claims

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Application Information

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IPC IPC(8): A61K39/395A61P35/00A61K31/7084
Inventor 张跃茹袁红谭瀛轩
Owner LIAOCHENG CITY ORIENT BIOMEDICAL TECH CO LTD
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