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A kind of lilrb4 and b7-h3 dual targeting chimeric antigen receptor and its application

A chimeric antigen receptor, B7-H3 technology, applied in the direction of targeting specific cell fusion, polypeptides containing positioning/targeting motifs, NGF-receptor/TNF-receptor superfamily, etc., can solve Hematopoietic system off-target toxicity and other issues, to achieve the effect of inhibiting tumor growth

Active Publication Date: 2022-05-27
CARBIOGENE THERAPEUTICS CO LTD
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  • Abstract
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Problems solved by technology

The reason is mainly that the above-mentioned targets are expressed to varying degrees in normal hematopoietic progenitor cells, so the continuous persistence of CAR-T cells in the body may cause irreversible off-target toxicity to the hematopoietic system

Method used

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  • A kind of lilrb4 and b7-h3 dual targeting chimeric antigen receptor and its application
  • A kind of lilrb4 and b7-h3 dual targeting chimeric antigen receptor and its application
  • A kind of lilrb4 and b7-h3 dual targeting chimeric antigen receptor and its application

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Embodiment 1

[0065] 1) Determination of LILRB4-B7H3-CAR-IFNα2b T gene sequence and construction of retroviral vector:

[0066] The scFv sequence of LILRB4 was derived from clone #293623; the scFv sequence of B7H3 was derived from clone #C11D5.3. The full-length cDNA sequence information of human CD8 hinge transmembrane region, human 4-1BB intracellular region, human CD3ζ intracellular region and human IFNα2b was searched from the NCBI website database. The full-length cDNA sequence of the wild-type human IFNα2b gene is called nIFNα2b. The nIFNα2b sequence is codon-optimized on the website http: / / sg.idtdna.com / site to obtain oIFNα2b, which ensures that the encoded amino acid sequence is more suitable for human cell expression.

[0067] According to the sequence of LILRB4 scFv, linker, B7-H3 scFv, human CD8 hinge transmembrane region, human 4-1BB intracellular region, human CD3ζ intracellular region, P2A peptide, oIFNα2b, LILRB4-B7H3-CAR-IFNα2b full-length polynucleosides were obtained aci...

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Abstract

The invention relates to the field of chimeric antigen receptors and discloses a chimeric antigen receptor. The chimeric antigen receptor comprises anti-LILRB4 single-chain antibody, anti-B7-H3 single-chain antibody, human CD8 hinge transmembrane region, human 4-1BB intracellular region, human CD3ζ intracellular region, human P2A peptide and Human IFN full length. The results of cytological experiments show that T cells expressing the chimeric antigen receptor have a better killing effect on tumor cells than single-target CAR-T of the same type. The results of animal experiments show that T cells expressing the chimeric antigen receptor can significantly prolong the survival rate of animals, and the effect is better than that of single-target CAR-T of the same type. The invention also discloses the application of a LILRB4 and B7-H3 double-targeted chimeric antigen receptor for preparing a medicine for treating acute myeloid leukemia.

Description

technical field [0001] The present invention relates to the technical field of chimeric antigen receptors, in particular to a chimeric antigen receptor with dual targeting of LILRB4 and B7-H3 and its application. Background technique [0002] Chimeric antigen receptor T (CAR-T) cell therapy is emerging as a promising immunotherapy strategy due to its remarkable efficacy in hematological tumors. CAR-T cells refer to T cells that have been artificially engineered to express a chimeric antigen receptor (CAR) on the cell surface. CAR is the core component of CAR-T, which endows T cells with the ability to recognize tumor-associated antigens (TAAs) on the tumor surface in a histocompatibility antigen (HLA)-independent manner. Therefore, CAR-T cells can specifically target tumor cells expressing TAA surface markers in vivo, and kill tumor cells by activating the T cell immune response. In 2010, the first patient with B-cell lymphoma received CD19-targeting CAR-T cell therapy wit...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00C12N15/62C12N5/10A61K39/00A61P35/02
CPCC07K16/2803C07K16/2827C07K14/7051C07K14/56C07K14/565C07K14/70517C07K14/70578C12N5/0636A61K39/001141A61K39/001112A61K39/001111A61P35/02C07K2317/622C07K2319/02C07K2319/03C07K2319/33C07K2319/74C12N2510/00A61K2039/5156A61K2039/804
Inventor 朱建高杨文君
Owner CARBIOGENE THERAPEUTICS CO LTD
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