Recombinant oncolytic vaccinia virus and preparation method and application thereof

A vaccinia virus and oncolytic technology, applied in the field of genetic engineering, can solve the problems of serious immune-related side effects, patients cannot get a complete response, and affect the efficacy of PD-1 monoclonal antibody, and achieve the effect of enhancing killing ability and enhancing safety.

Inactive Publication Date: 2018-06-15
ZHEJIANG UNIV
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Problems solved by technology

[0007] However, immune checkpoint inhibitors also have their limitations: (1) Most patients cannot achieve a durable complete response; (2) About 25-30% of patients experience severe immune-relat

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  • Recombinant oncolytic vaccinia virus and preparation method and application thereof
  • Recombinant oncolytic vaccinia virus and preparation method and application thereof
  • Recombinant oncolytic vaccinia virus and preparation method and application thereof

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Embodiment Construction

[0025] The present invention is further described in conjunction with accompanying drawings and examples. Unless otherwise specified, the present invention may adopt conventional techniques in the art.

[0026] 1. Synthesis of human full-length PD-1 monoclonal antibody:

[0027] The sequence of the single-chain variable region of the PD-1 monoclonal antibody refers to the sequence of Bristol-Myers Squibb’s PD-1 monoclonal antibody nivolumab, and the sequence of the PD-1 single-chain variable region designed by us is obtained through amino acid deduction and codon optimization. The Fc segment of the monoclonal antibody Referring to the Fc segment sequence of human IgG4, the heavy chain and light chain of human PD-1 antibody were synthesized by chemical synthesis, and the heavy chain and light chain genes were linked by the linker IRES. The full-length gene sequence of the PD-1 monoclonal antibody is shown in SEQ ID NO.1.

[0028] 2. Construction and identification of oncolyti...

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Abstract

The invention discloses a recombinant oncolytic vaccinia virus and a preparation method and application thereof. The thymidine kinase (TK) region of the virus comprises a coding sequence of a PD1 full-length antibody shown in the formula of SEQ ID NO.1. Through effective combination of the anti-tumor effect of gene therapy and the oncolytic effect of virus treatment, the oncolytic vaccinia virus for efficiently expressing the PD1 full-length antibody gene is prepared. When the oncolytic vaccinia virus produces the oncolytic effect of splitting tumor cells, the PD1 full-length antibody is efficiently expressed, the activity of PD1 on the surfaces of T cells is inhibited, the T cell immune response is activated and the dual antitumor effects are produced. Compared with the single gene therapy or viral therapy, the method using the recombinant oncolytic vaccinia virus improves the effects of killing malignant B cell lymphoma. Through virus replication-related gene deletion, the TK regionof the vaccinia virus genome is deleted so that specific replication of the viruses in the abnormally proliferating tumor cells is ensured and the viruses can not replicate in normal cells, so that the safety of the oncolytic vaccinia virus vector is greatly improved.

Description

technical field [0001] The invention belongs to genetic engineering, immunology and oncology, and relates to a recombinant oncolytic vaccinia virus and its preparation method and application. Background technique [0002] The incidence of malignant lymphoma accounts for 3-5% of all cancer patients, with 452,000 new patients every year. More than 90% of adult patients are derived from mature B cells, suggesting that B cell lymphoma is one of the important diseases that threaten human health. [0003] Immunotherapy is another new method of cancer treatment after surgery, radiotherapy, chemotherapy, and molecular targeted therapy. Among various cancer immunotherapy methods, immune checkpoint inhibitors are one of the drugs that have achieved outstanding efficacy in recent years. Immune checkpoints are inhibitory pathways that prevent excessive activation of the immune system. There are some proteins such as CTLA4 and PD1 on the surface of activated immune cells that act as "b...

Claims

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Application Information

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IPC IPC(8): C12N7/01A61K48/00A61K35/768A61P35/00
CPCA61K35/768A61K48/005C07K16/2818C12N7/00C12N2710/24121C12N2740/16034
Inventor 钱文斌雷文王世兵
Owner ZHEJIANG UNIV
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