Anti-B7H3 chimeric antigen receptor and application thereof

A technology of chimeric antigen receptor and antigen, applied in the field of biomedicine

Active Publication Date: 2021-03-19
GUANGZHOU BIO GENE TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few reports on immunotherapy targeting B7H3

Method used

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  • Anti-B7H3 chimeric antigen receptor and application thereof
  • Anti-B7H3 chimeric antigen receptor and application thereof
  • Anti-B7H3 chimeric antigen receptor and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0131] Example 1 Preparation of CAR-T cells

[0132] In this example, anti-B7H3 antibodies H26B6, H2B8, 26B6, 2B8, 23H1, 6F7, Enoblituzumab (Eno) and huM30 were selected as antigen-binding domains to construct CAR molecules, among which, 26B6 and its humanization H26B6, 2B8 and its humanization H2B8, 23H1, and 6F7 have significant binding ability to B7H3, not only can bind to free B7H3 protein, but also can bind to B7H3 protein on the cell surface; huM30 is a humanized B7H3 antibody (CN103687945B) of Daiichi Sankyo, Japan ), is conducting phase I clinical trials for the treatment of B7H3-positive solid tumors (NCT02192567); Enoblituzumab (MGA271) is a novel monoclonal antibody targeting B7H3 that has been optimized by immune molecules, and is developed by MacroGenics using exclusive Fc optimization technology Enoblituzumab has unique antibody advantages and therapeutic potential. There is no such drug approved in the world. Enoblituzumab represents the world's leading B7H3 ant...

Embodiment 2

[0149] Example 2 CAR-T cell expression efficiency of CAR

[0150] The expression rate of CAR in CAR-T cells was detected by flow cytometry.

[0151] Such as Figure 3A As shown, the expression rate of CAR in H26B6-CAR-T cells was 65.72%, and the expression rates of CAR in H2B8-CAR-T, L2B8-CAR-T, and L26B6-CAR-T cells were 31.73%, 38.15%, and 44.14%, respectively.

[0152] Such as Figure 3B As shown, the expression rate of CAR in Eno-CAR-T cells was 27.3%, and the expression rate of CAR in huM30-CAR-T cells was 45.2%.

[0153] In another experiment, as Figure 3C As shown, the expression rate of CAR in huM30-CAR-T cells was 23.09%, and the expression rate of CAR in 26B6-CAR-T cells was 7.67%;

[0154] In another experiment, as Figure 3D As shown, the CAR expression rate of huM30-CAR-T cells was 33.12%, that of L2B8-CAR-T cells was 33.43%, and that of 2B8-CAR-T cells was 12.55%.

Embodiment 3

[0155] Example 3 Killing function of CAR-T cells

[0156] H26B6-CAR-T, H2B8-CAR-T, L2B8-CAR-T, L26B6-CAR-T were mixed with human liver cancer cell HepG2, human pancreatic cancer cell PL45, and human cervical cancer cell SiHa at 2:1, 1:1 , 1:4 effect-to-target ratio and incubated for 16 hours, using RTCA technology to detect the killing efficiency of CAR-T.

[0157] Figure 4A , Figure 4B , Figure 4C The results showed that the four CAR-T cells had killing effects on the three tumor cells under different effect-to-target ratios, and the greater the effect-to-target ratio, the stronger the killing ability; when the effect-to-target ratio was 2:1, H26B6-CAR-T The killing efficiency of T on the three tumor cells was better than that of H2B8-CAR-T, L2B8-CAR-T and L26B6-CAR-T.

[0158] Different CAR-T cells (huM30-CAR-T and 2B8-CAR-T) were prepared using PBMCs from healthy donors (donor 1 and donor 2), respectively, and the target cells were mixed at a ratio of 3:1, 1:1, and 1...

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Abstract

The invention provides an anti-B7H3 chimeric antigen receptor and application thereof. The anti-B7H3 chimeric antigen receptor comprises an antigen binding domain, a hinge region, a transmembrane domain and a signal transduction domain, wherein the antigen binding domain is an anti-human B7H3 antibody. The anti-B7H3 chimeric antigen receptor has a specific targeting effect on B7H3 positive tumor cells, T cells expressing the anti-B7H3 chimeric antigen receptor have a remarkable in-vitro and in-vivo killing effect, the B7H3 positive tumor cells can be effectively removed, and the anti-B7H3 chimeric antigen receptor has important significance in the field of tumor treatment.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and relates to an anti-B7H3 chimeric antigen receptor and its application. Background technique [0002] B7H3 is a type I transmembrane protein, which belongs to the B7 immune co-stimulatory and co-inhibitory family. It has immunosuppressive function, can reduce the type I interferon (IFN) released by T cells, and reduce the cytotoxicity of NK cells. B7H3 protein has limited expression in normal tissues (such as prostate, breast, placenta, liver, colon and lymphoid organs), but abnormally high expression in most malignant tumors. The expression of B7H3 can be detected in non-small cell lung cancer cell lines and tumor tissues. In B7H3-expressing tumor tissues, the number of infiltrating lymphoid cells was significantly reduced and positively correlated with lymph node metastasis (Sun Y, Wang Y, Zhao J, et al. B7-H3 and B7-H4 expression in non-small-cell lung cancer[J].Lung Cancer,2006,53(2)...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N15/62C12N15/867C12N5/10C12N5/0783A61K39/00A61P35/00A61P35/02
CPCC07K16/2827C07K14/7051C12N5/0636C12N15/86A61K39/001111A61P35/00A61P35/02C07K2319/02C07K2319/03C12N2510/00C12N2740/15043A61K2039/5158C07K14/70517C07K14/70521C07K2317/622C07K14/70578A61K39/4611A61K39/4631A61K39/464411C07K2317/24
Inventor 罗敏李光超周兆王学俊
Owner GUANGZHOU BIO GENE TECH CO LTD
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