Cyclic mRNA (messenger ribonucleic acid) tumor immune drug for colorectal cancer

A colorectal cancer and tumor technology, applied in the field of potential colorectal cancer tumor immune drugs, to achieve the effect of reducing social security pressure

Pending Publication Date: 2022-04-29
奥明(杭州)生物医药有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, colorectal cancer is not a single disease and is highly heterogeneous. There is an urgent clinical need for effective immune drugs that can be used for the prevention and treatment of colorectal cancer, so as to improve the survival rate and quality of life of patients

Method used

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  • Cyclic mRNA (messenger ribonucleic acid) tumor immune drug for colorectal cancer
  • Cyclic mRNA (messenger ribonucleic acid) tumor immune drug for colorectal cancer
  • Cyclic mRNA (messenger ribonucleic acid) tumor immune drug for colorectal cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0116] Example 1 Expression of circular mRNA mediated by IRES in 293T cells

[0117] The first step is to construct a regulatory sequence (SEQ ID 52), a 5´ homology arm (SEQ ID 53), a 3´ intron (SEQID 54), a second exon (SEQ ID 55), and a 5´ spacer (SEQ ID 56), the IRES element with the nucleotide sequence shown in SEQ ID 51, EGFP coding region (SEQ ID 57), 3' spacer (SEQ ID 58), the first exon (SEQ ID59), 5 ´ intron (SEQ ID 60), 3´ homology arm (SEQ ID 61), and the restriction site XbaI (SEQ ID 62) that can be used for plasmid linearization to express the target gene of the antigen polypeptide, and connect the resulting gene fragments to the pUC57 vector;

[0118] In the second step, after activating the synthetic puncture bacteria at 36.8 degrees Celsius and 200 rpm for 4 hours, take the activated bacteria liquid and cultivate it overnight at 36.8 degrees Celsius and 200 rpm. Then, the plasmid was extracted, and the OD value and plasmid extraction concentration (342.1 ng p...

Embodiment 2

[0126] Example 2 Expression of circular mRNA mediated by IRES in 293T cells

[0127] The first step is to construct a regulatory sequence (SEQ ID 52), a 5´ homology arm (SEQ ID 53), a 3´ intron (SEQID 63), a second exon (SEQ ID 55), and a 5´ spacer (SEQ ID 56), the IRES element with the nucleotide sequence shown in SEQ ID 51, EGFP coding region (SEQ ID 57), 3' spacer (SEQ ID 58), the first exon (SEQ ID59), 5 ´ intron (SEQ ID 60), 3´ homology arm (SEQ ID 61), and the restriction site XbaI (SEQ ID 62) that can be used for plasmid linearization to express the target gene of the antigen polypeptide, and connect the resulting gene fragments to the pUC57 vector;

[0128] In the second step, after activating the synthetic puncture bacteria at 36.8 degrees Celsius and 200 rpm for 4 hours, take the activated bacteria liquid and cultivate it overnight at 36.8 degrees Celsius and 200 rpm. Then, the plasmid was extracted, and the OD value and plasmid extraction concentration (282.1 ng p...

Embodiment 3

[0136] Example 3 Expression of circular mRNA in 293T cells mediated by IRES

[0137] The first step is to construct a regulatory sequence (SEQ ID 52), a 5´ homology arm (SEQ ID 53), a 3´ intron (SEQ ID 63), a second exon (SEQ ID 64), and a 5´ spacer (SEQ ID 56), the IRES element with the nucleotide sequence shown in SEQ ID 51, EGFP coding region (SEQ ID 57), 3' spacer (SEQ ID 58), the first exon (SEQ ID59), 5 ´ intron (SEQ ID 60), 3´ homology arm (SEQ ID 61), and the restriction site XbaI (SEQ ID 62) that can be used for plasmid linearization to express the target gene of the antigen polypeptide, and connect the resulting gene fragments to the pUC57 vector;

[0138] In the second step, after activating the synthetic puncture bacteria at 36.8 degrees Celsius and 200 rpm for 4 hours, take the activated bacteria liquid and cultivate it overnight at 36.8 degrees Celsius and 200 rpm. Then, the plasmid was extracted, and the OD value and plasmid extraction concentration (302.4 ng ...

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Abstract

The invention discloses a potential colorectal cancer tumor immune drug. The colorectal cancer tumor immune drug is based on a colorectal cancer tumor neoantigen and a circular mRNA molecule with a specific IRES sequence. The colorectal cancer tumor immune drug is expected to expand the benefit range of colorectal cancer treatment patients, improve the survival rate of the colorectal cancer patients, and bring more colorectal cancer treatment benefits to the colorectal cancer patients, patient families and society.

Description

technical field [0001] The present invention relates to a class of potential colorectal cancer tumor immune drugs, in particular to a class of circular mRNA with a specific IRES sequence based on colorectal cancer tumor neoantigens and capable of expressing the colorectal cancer neoantigens in human T cells Molecular tumor immune drugs. Background technique [0002] Colorectal cancer is the third leading cause of cancer death worldwide. In China, in 2020, colorectal cancer has surpassed gastric cancer to become the second most common cancer. In addition, nearly 80% of colorectal cancer patients have reached the middle and advanced stages when they are discovered, and nearly half of the patients survive less than 5 years. However, colorectal cancer is not a single disease with high heterogeneity. There is an urgent clinical need for effective immune drugs that can be used for the prevention and treatment of colorectal cancer, so as to improve the survival rate and quality o...

Claims

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Application Information

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IPC IPC(8): C07K14/47C12N15/12C12N15/85C12N5/10A61K39/00A61P35/00
CPCC07K14/4748C12N15/85A61K39/0011A61P35/00C12N2800/107A61K2039/82A61K2039/836
Inventor 童云广钱艳王琼杨开珍张克海徐玉莲
Owner 奥明(杭州)生物医药有限公司
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