Chimeric receptors and methods of use thereof

Abstract

This article provides information on chimeric receptor-targeted acute myeloid leukemia antigens and their application.

Description

Prior Technology

Content of invention

Implementation

Claims

1. An isolated immune-responding cell comprising: (a) a first chimeric receptor comprising an extracellular antigen-binding domain binding to a first antigen, and a second chimeric receptor comprising an extracellular antigen-binding domain binding to a second antigen; or (b) a chimeric receptor comprising two or more antigen-binding domains, wherein each of the two or more antigen-binding domains binds to an antigen, wherein each antigen-binding domain binds to a different antigen, and wherein the two or more antigen-binding domains comprise at least an extracellular antigen-binding domain binding to the first antigen. The first antigen is FLT3 and the second antigen is CD33; wherein the extracellular antigen-binding domain binding to the first antigen includes a complementarity-determining region (CDR)-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3, wherein CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively contain the following amino acid sequences: (a) SEQ (a) SEQ ID NO: amino acid sequences 81 to 86; (b) SEQ ID NO: amino acid sequences 75 to 80; (c) SEQ ID NO: amino acid sequences 87 to 92; (d) SEQ ID NO: amino acid sequences 93 to 98; (e) SEQ ID NO: amino acid sequences 99 to 104; (f) SEQ ID NO: amino acid sequences 105 to 110; (g) SEQ ID NO: amino acid sequences 111 to 116; or (h) SEQ ID NO: amino acid sequences 81 to 86. The amino acid sequences NO: 117 to 122; and the extracellular antigen-binding domain bound to the second antigen includes CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3, wherein CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 each contain the following amino acid sequences: (a) amino acid sequences SEQ ID NO: 123 to 128; or (b) amino acid sequences SEQ ID NO: 129 to 134.

2. The isolated immunoreactive cell of claim 1, wherein the extracellular antigen-binding domain bound to the first antigen comprises a heavy chain variable domain (VH) and a light chain variable domain (VL) selected from the group consisting of: (a) a VH comprising the amino acid sequence of SEQ ID NO: 3 and a VL comprising the amino acid sequence of SEQ ID NO: 4; (b) a VH comprising the amino acid sequence of SEQ ID NO: 1 and a VL comprising the amino acid sequence of SEQ ID NO: 2; (c) a VH comprising the amino acid sequence of SEQ ID NO: 5 and a VL comprising the amino acid sequence of SEQ ID NO: 6; (d) a VH comprising the amino acid sequence of SEQ ID NO: 7 and a VL comprising the amino acid sequence of SEQ ID NO: 8; (e) a VH comprising the amino acid sequence of SEQ ID NO: 9 and a VL comprising the amino acid sequence of SEQ ID NO: 10; (f) comprising the amino acid sequence of SEQ ID NO:

3. (g) VH containing the amino acid sequence of SEQ ID NO: 11 and VL containing the amino acid sequence of SEQ ID NO: 12; and (h) VH containing the amino acid sequence of SEQ ID NO: 13 and VL containing the amino acid sequence of SEQ ID NO: 14; and (h) VH containing the amino acid sequence of SEQ ID NO: 15 and VL containing the amino acid sequence of SEQ ID NO:

16.

3. The isolated immune response cells of claim 2, wherein the extracellular antigen-binding domain bound to the second antigen comprises a heavy chain variable domain (VH) and a light chain variable domain (VL) selected from the group consisting of: (a) a VH comprising the amino acid sequence of SEQ ID NO: 17 and a VL comprising the amino acid sequence of SEQ ID NO: 18, and (b) a VH comprising the amino acid sequence of SEQ ID NO: 19 and a VL comprising the amino acid sequence of SEQ ID NO:

20.

4. The isolated immune-responding cells as claimed in any of claims 1 to 3, wherein each chimeric receptor is a chimeric antigen receptor (CAR), and each CAR comprises: i) an intracellular signal transduction domain of the CD3ζ chain; and / or ii) a transmembrane domain selected from the group consisting of: CD8 transmembrane domain, CD28 transmembrane domain, CD3ζ chain transmembrane domain, CD4 transmembrane domain, 4-1BB transmembrane domain, OX40 transmembrane domain, ICOS transmembrane domain, CTLA-4 transmembrane domain, PD-1 transmembrane domain, LAG-3 transmembrane domain, 2B4 transmembrane domain, and BTLA transmembrane domain; and / or iii) a spacer region located between the antigen-binding domain and the transmembrane domain, and the spacer region having an amino acid sequence selected from the group consisting of SEQ ID NO: 55-64.

5. The isolated immune response cells as claimed in claim 4, wherein each CAR further comprises one or more additional intracellular signal transduction domains, and the one or more additional intracellular signal transduction domains are selected from the group consisting of: CD97 intracellular signal transduction domain, CD11a-CD18 intracellular signal transduction domain, CD2 intracellular signal transduction domain, ICOS intracellular signal transduction domain, CD27 intracellular signal transduction domain, CD154 intracellular signal transduction domain, CD8 intracellular signal transduction domain, OX40 intracellular signal transduction domain, 4-1BB intracellular signal transduction domain, CD28 intracellular signal transduction domain, ZAP40 intracellular signal transduction domain, CD30 intracellular signal transduction domain, GITR intracellular signal transduction domain, HVEM intracellular signal transduction domain, DAP10 intracellular signal transduction domain, DAP12 intracellular signal transduction domain, MyD88 intracellular signal transduction domain, and 2B4 intracellular signal transduction domain.

6. The isolated immune response cells of any one of claims 1 to 3, wherein the cells further comprise an inhibitory chimeric receptor comprising an antigen-binding domain.

7. The isolated immune response cells as claimed in claim 6, wherein the inhibitory chimeric receptor binds to antigens expressed on non-tumor cells.

8. The isolated immune response cells as claimed in claim 6, wherein the inhibitory chimeric receptor binds to antigens selected from the group consisting of: EMCN, JAM2, MS4A15, C4BPA, TRPM1, SCTR, SLC2A2, KCNQ2, PEP, WLS, FFAR2, PTPRB, NCKAP1, MPZL2, PLSCR4, TMEM47, ADGRL4, MET, BACE2, ATP8B1, LIFR, ART4, CALCRL, CNTNAP3, PCDH9, IL18R1, SLC8A3, CDH26, TMEM163, ABCA13, CACHD1, CYYR1, ABCB1, ADGRG6, ATP9A, CALN1, CDCP1, IL12RB2, SLC16A14, TMEM136, and TMEM200A.

9. The isolated immune response cells of claim 6, wherein the inhibitory chimeric receptor comprises an antigen-binding domain comprising a single-chain variable fragment (scFv) derived from an anti-EMCN antibody.

10. The isolated immune response cells of claim 6, wherein the antigen-binding domain of the first chimeric receptor, the antigen-binding domain of the second chimeric receptor and / or the antigen-binding domain of the inhibitory chimeric receptor comprises one or more single-chain variable fragments (scFv), wherein each of the one or more scFv comprises a heavy chain variable domain (VH) and a light chain variable domain (VL).

11. The isolated immune response cells of claim 10, wherein the VH and the VL are separated by peptide linkers.

12. The isolated immune response cells of claim 11, wherein the peptide linker contains the amino acid sequence of SEQ ID NO:

27.

13. The isolated immune response cells of claim 10, wherein each of the one or more scFvs binds to a different antigenic determinant on the same antigen.

14. The isolated immune response cells as claimed in claim 10, wherein each of the one or more scFvs is separated by peptide linkers.

15. The isolated immune response cells of claim 14, wherein the peptide linker contains the amino acid sequence GGGGSGGGGSGGGGS (SEQ ID NO: 27) or EAAAKEAAAKEAAAKEAAAK (SEQ ID NO: 74).

16. Isolated immune response cells as requested in any of items 1 to 3, wherein the cell line is selected from the following groups: T cells, natural killer (NK) cells, cytotoxic T lymphocytes (CTL), regulatory T cells, natural killer T (NKT) cells, bone marrow cells, macrophages, human embryonic stem cells (ESC), ESC-derived cells, pluripotent stem cells and induced pluripotent stem cells (iPSC), and iPSC-derived cells.

17. The isolated immune response cells of claim 16, wherein the immune response cells are allogeneic or autologous.

18. A pharmaceutical composition comprising an effective amount of any one of claims 1 to 17 isolated immune-responding cells and a pharmaceutically acceptable carrier, a pharmaceutically acceptable excipient, or a combination thereof.

19. Use of an isolated immune-response cell as claimed in any one of claims 1 to 17 or a pharmaceutical composition as claimed in claim 18, for the preparation of a medicament that provides antitumor immunity in an individual.

20. Use of an isolated immune-response cell as claimed in any one of claims 1 to 17 or a pharmaceutical composition as claimed in claim 18, for the preparation of a medicament for the treatment or prevention of bone marrow disorders in an individual.

21. As claimed in claim 20, wherein the bone marrow disease is myelodysplastic syndrome, myeloproliferative neoplasm, chronic myelomonocytic leukemia, acute myeloid leukemia (AML), acute myeloblastic leukemia, acute promyelocytic leukemia, acute myelomonocytic leukemia, chronic myelocytic leukemia, and polycythemia vera.

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