A fourth-generation car-t cell and its construction method and application

A technology of XCAR1&CAR2-Y-Z and cells, applied in the field of biotechnology engineering, can solve the problems of lack of target antigen, short duration of CAR-T cells, and poor effect of CAR-T cells in treating solid tumors, so as to reduce the recurrence of malignant tumors, Effect of promoting T cell proliferation and maintaining T cell stability

Active Publication Date: 2021-08-06
温州启星生物技术有限公司
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

In recent years, the achievements of CAR-T therapy in the treatment of malignant hematological tumors are obvious to all (such as Kymriah for refractory / relapsed acute B-lymphoblastic leukemia and refractory / relapsed non-Hodgkin's Lymphoma’s Yescarta was launched in the United States last year), but at least 50% of the recurrence rate, and CAR-T cells are not effective in treating solid tumors, mainly due to the lack of suitable target antigens and the persistence of CAR-T cells in vivo Challenging problems such as short time, immune escape, and immunosuppressive tumor microenvironment

Method used

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  • A fourth-generation car-t cell and its construction method and application
  • A fourth-generation car-t cell and its construction method and application
  • A fourth-generation car-t cell and its construction method and application

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Embodiment Construction

[0043] The embodiments of the present invention will be described in detail below with reference to the accompanying drawings, but the present invention can be implemented in many different ways defined and covered by the claims.

[0044] Main experimental materials:

[0045] EcoRI-HF, MluI-HF, NdeI restriction endonuclease (NEB Company), Seamless Cloning Enzyme (He Yuan Biotechnology), High Fidelity Prime GXL STAR Enzyme (TAKARA Company), TransStbl3 Competent Cells (Quanshijin Biotechnology Ltd.), Plasmid MiniKitI (OMEGA), Plasmid MaxiKit (QIAGEN), DMEM, RPMI-1640, Opti-MEM medium, GibcoFBS (Thermo Fisher Scientific), Sanger sequencing (Shanghai Sunny Biological Co., Ltd.), Nacl, yeast powder, peptone, EDTA, NaOH (Shanghai Sangong Biological Engineering Co., Ltd.), primers (Jiangsu Jinweizhi Biotechnology Co., Ltd.).

[0046] 1. Construction of recombinant plasmids

[0047] (1) Construction of Plenti-Nectin4 CAR1 and Plenti-Nectin4 CAR2 recombinant plasmids:

[0048] Ne...

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Abstract

This application provides the construction and application of a fourth-generation chimeric antigen receptor T (CAR-T) cell (expressing IL-7 and CCL19) targeting Nectin-4 on the surface of malignant tumors. The present invention takes the two spatial epitopes of the Nectin-4 antigen as targets, and the fourth-generation CAR-T constructed is used to treat malignant solid tumors expressing the Nectin-4 antigen, so as to solve the immune problem of CAR-T in the treatment of solid tumors escape problem; and the construction of the fourth-generation CAR-T can enhance its proliferation ability and continuous survival ability, so as to effectively treat solid tumors, and provide a new strategy for the effective prevention and treatment of solid tumor recurrence / metastasis after surgery.

Description

technical field [0001] The invention belongs to the field of biotechnology engineering, and in particular relates to the construction and application of the fourth-generation chimeric antigen receptor T (CAR-T) cells (expressing IL-7 and CCL19) aimed at the dual targets of Nectin-4 on the surface of malignant tumors. Background technique [0002] With the continuous deterioration of global environmental factors, the incidence and mortality of malignant tumors are on the rise, regardless of developed or developing countries, and malignant tumors have become the greatest threat to human life and health. About 4.3 million people in China were diagnosed with cancer in 2015, and 2.8 million people died of cancer, most of which were solid tumors (such as lung cancer, breast cancer, liver cancer, gastric cancer, colorectal cancer, bladder cancer, etc.). There will be 5.5 million new cancer cases and 4 million deaths. Due to the inherent defects of traditional treatment methods and...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/10C12N15/867A61K35/17A61P35/00
CPCA61K35/17A61P35/00C07K14/52C07K14/5418C07K14/7051C07K14/70517C07K16/2803C07K2317/622C07K2319/00C07K2319/02C12N5/0636C12N15/86C12N2510/00C12N2740/15043
Inventor 高基民李帆帆
Owner 温州启星生物技术有限公司
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