Fusion protein, coding nucleic acid, cell and application

A technology of fusion protein and nucleic acid, which is applied in the field of immunosuppressants to relieve immunosuppression, can solve the problems of slow disease progression and achieve the effect of enhancing anti-tumor function

Active Publication Date: 2020-04-03
启辰生生物科技(珠海)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, the use of drugs targeting CD223 and pembrolizumab (PD-1 monoclonal antibody) phase I clinical dose-escalation treatment of unresectable or metastatic melanoma has been publi

Method used

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  • Fusion protein, coding nucleic acid, cell and application
  • Fusion protein, coding nucleic acid, cell and application
  • Fusion protein, coding nucleic acid, cell and application

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0043] The preparation method of the cell of the present invention is not particularly limited. In an exemplary preparation method, the preparation method of the cell of the present invention includes the following steps:

[0044] (1) Construct nucleic acid capable of producing fusion protein;

[0045] (2) Separate peripheral blood mononuclear cells from venous blood, and induce differentiation to obtain antigen-presenting cells; and

[0046] (3) The nucleic acid of step (1) is introduced into the antigen-presenting cell of step (2), and the antigen-presenting cell is cultured under conditions suitable for expression of the nucleic acid.

[0047] In certain embodiments, the method of the present invention includes preparing a plasmid containing the corresponding DNA encoding. After that, the in vitro transcription process is performed. First, the plasmid is linearized with restriction endonuclease, and the linearized plasmid is used as a template to prepare ribonucleic acid molecules...

Example Embodiment

[0055] Example 1

[0056] This preparation example is a method for preparing an exemplary fusion protein.

[0057] 1. Preparation of DNA and mRNA constructs

[0058] The gene used to generate an exemplary fusion protein of the present invention was constructed, and its sequence is shown in SEQ ID No.5. In addition, the genes used to generate separate soluble immunoglobulin CD223 and soluble programmed death receptor 1 as controls were further constructed, the sequences of which are shown in SEQ ID No. 6 and 7, respectively. In addition, the antigen GPC3 used in subsequent experiments is encoded by the sequence shown in SEQ ID No.8. The sequence information is shown in Table 1 below.

[0059] Table-1 DNA sequence list

[0060] name serial number Fusion protein SEQ ID No. 5 Soluble immunoglobulin CD223 SEQ ID No. 6 Soluble programmed death receptor 1 SEQ ID No. 7 GPC3 SEQ ID No. 8

[0061] 2. In vitro transcription

[0062] First, use restriction endonuclease to linearize the p...

Example Embodiment

[0074] Example 2

[0075] The experiment was conducted in the same manner as in Example 1, except that the antigen in Example 1 was changed to AFP. The results are shown in the table below.

[0076]

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Abstract

The invention discloses a fusion protein, a coding nucleic acid, a cell and application. The fusion protein of the present invention comprises a soluble immunoglobulin CD223 as a first part and a soluble programmed death receptor 1 as a second part. The first part is covalently linked to the second part through a linker. The fusion protein disclosed by the invention has a specific Y-shaped structure, so that the functions of the first part and the second part have a synergistic effect, a PD-1 signal channel can be blocked, and an LAG3 signal channel can be blocked, so that inhibition of the PD-1/LAG3 signal channel on T cells after activation can be effectively relieved, and the anti-tumor function of the T cells is remarkably enhanced.

Description

technical field [0001] The present invention relates to the fields of immunology and medicine, in particular to a fusion protein used to block the immunosuppressive site of tumor cells, its encoding nucleic acid, and cells containing such fusion protein or nucleic acid, and as an immunosuppressant in releasing immunosuppression use. Background technique [0002] Since the first PD-1 antibody drug was launched on the market in 2015, this type of drug has shown excellent safety and efficacy. At present, combined drugs based on PD-1 and PDL-1 have become a new direction for tumor treatment, and some have been developed. Exciting data have also been obtained in the clinical combination drug research, and more and more people have confidence in the combination drug of immune checkpoints. [0003] PD-1 inhibitors include PD-1 antibodies and PD-L1 antibodies, and are a new drug for tumor immunotherapy. Unlike surgery, radiotherapy and chemotherapy, and targeted drugs, PD-1 inhibi...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N15/62C12N5/10A61K39/395A61P35/00A61K38/17
CPCA61K38/177A61K39/39558A61P35/00C07K14/705C07K14/70596C07K2319/00C12N5/0636C12N2510/00A61K2300/00
Inventor 蒋俊林鑫谢桦函
Owner 启辰生生物科技(珠海)有限公司
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