FUSION GENE CONTAINING THE ENCODING GENE OF A CHIMERIC ANTIGEN RECEPTOR AND THE ENCODING GENE OF A CHIMERIC SWITCH RECEPTOR AND ITS APPLICATION

Claims

1. A nucleic acid molecule that contains the encoding gene for a chimeric antigen receptor and the encoding gene for a chimeric switch receptor.

2. The nucleic acid molecule of claim 1, wherein the chimeric switch receptor comprises the extracellular region of the TGFβ (transforming growth factor beta) type II receptor, μPD-1 (mutation-optimized extracellular region of programmed cell death protein 1), the transmembrane region and the cytoplasmic region of CD27 (cluster of differentiation 27), and the amino acid sequence of μPD-1 corresponds to positions 187-332 of SEQ ID No.

1.

3. The nucleic acid molecule of claim 2, wherein the encoding gene for the chimeric switch receptor is one of the following: B1) a nucleic acid molecule encoding the fusion protein; B2) a DNA molecule whose coding sequence is SEQ ID No. 2; B3) a DNA molecule, the nucleotide sequence of which is SEQ ID No. 2; The fusion protein is one of the following: A1) a protein whose amino acid sequence is SEQ ID No. 1; A2) a protein having 80% or more identity with the protein specified in (A1) and having the same function, obtained by replacing and / or deleting and / or adding amino acid residues of the amino acid sequence shown in SEQ ID No. 1; A3) a fusion protein having the same function, obtained by attaching a label to the N-terminus and / or C-terminus of (A1) or (A2).

4. A nucleic acid molecule according to any one of claims 1-3, which further comprises an IFNα (interferon alpha) gene and / or a P2A (peptide 2A obtained from porcine teschovirus 1) gene.

5. The nucleic acid molecule of any one of claims 1 to 3, wherein the encoding gene for the chimeric antigen receptor is an encoding gene for a chimeric antigen receptor targeting GPC3 (glypican-3).

6. The nucleic acid molecule of claim 5, wherein the encoding gene for the chimeric antigen receptor targeting GPC3 is one of the following: C1) a nucleic acid molecule encoding a protein having the amino acid sequence SEQ ID No. 4; C2) a DNA molecule having a coding sequence from positions 1-1473 of SEQ ID No. 3; C3) a DNA molecule having a nucleotide sequence from positions 1-1473 of SEQ ID No.

3.

7. A nucleic acid molecule according to claim 6, which is any of the following: D1) a DNA molecule having the nucleotide sequence SEQ ID No. 3; D2) a DNA molecule having 70% or more identity with the DNA molecule indicated in (D1) and having the same function, obtained by modifying the nucleotide sequence, and / or replacing, and / or deleting, and / or adding one or more nucleotides indicated in SEQ ID No.

3.

8. A biomaterial that is any of the following: E1) an expression cassette containing a nucleic acid molecule according to any one of paragraphs 1-7; E2) a recombinant vector containing a nucleic acid molecule according to any one of claims 1 to 7, or a recombinant vector containing an expression cassette (E1); E3) a recombinant microorganism containing a nucleic acid molecule according to any one of paragraphs 1-7, or a recombinant microorganism containing an expression cassette (E1), or a recombinant microorganism containing a recombinant vector (E2); E4) a recombinant cell containing a nucleic acid molecule according to any one of claims 1 to 7, or a recombinant cell containing an expression cassette (E1), or a recombinant cell containing a recombinant vector (E2); E5) encoding the chimeric switch receptor vector gene according to claim 2 or 3; E6) chimeric switch receptor according to claim 2; E7) the fusion protein according to item 3.

9. The biomaterial according to claim 8, wherein the cell (E4) is a T cell, an NK cell (natural killer cell), a γδT cell (a T cell having a gamma-delta T cell receptor), an NKT cell (a T cell expressing NK cell markers and T cell differentiation antigens), a macrophage or a stem cell.

10. A nucleic acid molecule according to any of 1-7, and / or any of the following uses of the biomaterial according to claim 8 or 9: F1) use in the manufacture of a medicinal product for the prevention or treatment of tumors; F2) use in the manufacture of a medicinal product for the prevention or treatment of tumors expressing the GPC3 antigen; F3) use in regulating the immunosuppressive effect of the tumor microenvironment or in the manufacture of a product regulating the immunosuppressive effect of the tumor microenvironment; F4) use in the prevention or treatment of tumors; F5) use in the prevention or treatment of tumors expressing the GPC3 antigen; F6) use in the prevention or treatment of liver cancer, melanoma, Wilms' tumor, non-small cell lung cancer, clear cell ovarian cancer, squamous cell carcinoma, renal cell carcinoma, prostate cancer, colorectal cancer, hepatoblastoma or glioma.

11. A pharmaceutical composition, the active ingredient of which is a CAR (chimeric antigen receptor) cell containing or expressing any of the nucleic acid molecules of the present invention.

12. A method for preventing or treating tumors, which consists of administering a medicinal product containing a recombinant cell according to paragraph 8 to a patient suffering from a tumor disease.

13. The method of claim 12, wherein the recombinant cell is a GPC3 CAR-IC T cell, and the GPC3 CAR-IC T cell comprises the DNA molecule shown in SEQ ID No.

3.

14. The method according to claim 12 or 13, wherein the tumor is a tumor expressing the GPC3 antigen.

15. The method of claim 14, wherein the tumor expressing the GPC3 antigen is liver cancer, melanoma, Wilms tumor, non-small cell lung cancer, clear cell ovarian carcinoma, squamous cell carcinoma, renal cell carcinoma, prostate cancer, colorectal cancer, hepatoblastoma, or glioma.

16. The method of claim 15, wherein the liver cancer is hepatocellular carcinoma.

17. The method of claim 15, wherein the squamous cell carcinoma is squamous cell lung carcinoma.

18. The method of claim 15, wherein the melanoma is malignant melanoma.

19. The method according to claim 15, wherein the glioma is a glioblastoma.