JPWO2021162521A5

JPWO2021162521A5 -

Info

Publication number: JPWO2021162521A5
Application number: JP2022548955A
Authority: JP
Publication date: 2024-02-26

Claims

The following (i) to (v):
(i) antigen-binding domain;
(ii) transmembrane domain;
(iii) an intracellular signaling domain comprising at least one costimulatory domain;
(iv) a self-cleaving peptide; and (v) a polynucleotide encoding a fusion protein comprising a signal transduction pathway modulator.

2. The polynucleotide of claim 1, wherein a spacer is further placed between (i) the antigen-binding domain and (ii) the transmembrane domain.

The signal transduction pathway modulator is a) a protein located in the immunosuppressive signal transduction pathway, b) an immunophilin, c) a protein involved in antigen loss-mediated relapse, d) a protein located in the T cell stimulatory pathway, e) 2. The polynucleotide of claim 1, which is any one selected from the group consisting of: a protein involved in the inhibition of negative feedback; and f) a combination thereof.

2. The polynucleotide of claim 1, wherein the signal transduction pathway modulator functions in the cytoplasm.

a) The protein located in the immunosuppressive signal transduction pathway is a protein or fragment thereof located in the TGF-β/SMAD (FKBP12-FK506/rapamycin) signal transduction pathway , or a protein or fragment thereof located in the inhibitory immune checkpoint pathway. It is a fragment;
b) the immunophilin is FKBP12 or a fragment thereof, or cyclophilin A or a fragment thereof;
c) the protein involved in antigen loss-mediated relapse is a protein involved in trogocytosis, or a fragment thereof;
d) The protein located in the T cell stimulation pathway is an adapter of the TCR/Zap70 pathway or a fragment thereof, a protein located in the TNFR/TLR receptor (TRAF/NF-kB) pathway or a fragment thereof, a cytokine receptor (JAK-kB), or a protein located in the T cell stimulation pathway. a protein or a fragment thereof located in the MAP kinase pathway; or a protein located in the MAP kinase pathway or a fragment thereof;
e) the protein involved in negative feedback inhibition is a protein involved in negative feedback inhibition of a cytokine signaling pathway, or a fragment thereof ;
The polynucleotide according to claim 3.

The signal transduction pathway modulator has the following structural formula (I) or (II) :
N'-X-L1-Y-C' (I)
N'-X-L1-Y-L2-Z-C' (II)
In the structural formulas (I) and (II) ,
N' is the N-terminus of the fusion protein;
C' is the C-terminus of the fusion protein;
L1 is a self-cleaving peptide;
Each of X , Y and Z represents a) a protein or fragment thereof located in an immunosuppressive signaling pathway; b) an immunophilin or a fragment thereof; c) a protein or fragment thereof involved in antigen loss-mediated relapse; d) Any one selected from the group consisting of a protein or a fragment thereof located in the T cell stimulation pathway; and e) a protein or a fragment thereof involved in the inhibition of negative feedback.
The polynucleotide according to claim 1.

2. The polynucleotide of claim 1, wherein the signal transduction pathway modulator comprises FKBP12 protein or a fragment thereof, cyclophilin A, and the N-terminal SH2 domain of SHP2 protein or a fragment thereof.

The antigen-binding domain is alpha folate receptor, 5T4, αvβ6 integrin, BCMA, B7-H3, B7-H6, CAIX, CD16, CD19, CD20, CD22, CD30, CD33, CD44, CD44v6, CD44v7/8, CD70. , EGFR family including CD79a, CD79b, CD123, CD138, CD171, CEA, CSPG4, EGFR, and ErbB2 (HER2), EGFRvIII, EGP2, EGP40, EPCAM, EphA2, EpCAM, FAP, fetal AchR, FRα, GD2, G D3, Glypican-3 (GPC3), HLA-A1+MAGE1, HLA-A2+MAGE1, HLA-A3+MAGE1, HLA-A1+NY-ESO-1, HLA-A2+NY-ESO-1, HLA-A3+NY-ESO-1, IL-11Rα, IL-13Rα2 , lambda, Lewis-Y, kappa, mesothelin, Muc1, Muc16, NCAM, NKG2D ligand, NY-ESO-1, PRAME, PSCA, PSMA, ROR1, SSX, survivin, TAG72, TEM, VEGFR2, and WT-1. 2. The polynucleotide of claim 1, which specifically binds to any one antigen selected from the group.

The polynucleotide according to claim 1, wherein the antigen-binding domain consists of the amino acid sequence represented by SEQ ID NO:3.

The transmembrane domain is a T cell receptor, CD3δ, CD3ε, CD3γ, CD3ζ, CD4, CD5, CD8α, CD9, CD16, CD22, CD27, CD28, CD33, CD37, CD45, CD64, CD80, CD86, CD134, CD137 , CD152, CD154, AMN, and PD-1.

2. The polynucleotide of claim 1, wherein the transmembrane domain is derived from CD8α.

The polynucleotide according to claim 1, wherein the transmembrane domain consists of the amino acid sequence represented by SEQ ID NO:5.

2. The polynucleotide of claim 1, wherein the intracellular signaling domain comprises a costimulatory domain and a primary signaling domain.

The costimulatory domain includes TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, TLR10, CARD11, CD2, CD7, CD27, CD28, CD30, CD40, CD54 (ICAM), CD83, CD134 (OX40). ), CD137 (4-1BB), CD278 (ICOS), DAP10, LAT, NKD2C , SLP76, TRIM, and ZAP70.

14. The polynucleotide of claim 13 , wherein the costimulatory domain consists of the amino acid sequence represented by SEQ ID NO:7.

14. The polynucleotide of claim 13 , wherein the primary signaling domain is derived from FcRγ, FcRβ, CD3γ, CD3δ, CD3ε, CD3ζ, CD22, CD79a, CD79b, or CD66d.

14. The polynucleotide of claim 13 , wherein the primary signaling domain consists of the amino acid sequence represented by SEQ ID NO:9.

2. The polynucleotide of claim 1, wherein the self-cleaving peptide is derived from P2A, E2A, F2A, or T2A.

An expression vector comprising the polynucleotide according to claim 1 .

20. The vector of claim 19 , further comprising a sequence encoding a signal peptide.

21. The vector according to claim 20 , wherein the sequence encoding the signal peptide is inserted before the sequence encoding the antigen-binding domain.

A virus comprising the polynucleotide according to claim 1 .

An immune cell into which the polynucleotide according to claim 1 has been introduced.

The immune cell according to claim 23 , which is a T cell or a natural killer cell.

A pharmaceutical composition for treating or preventing cancer, comprising the immune cell according to claim 23 as an active ingredient.

The cancer is stomach cancer, liver cancer, lung cancer, colorectal cancer, breast cancer, prostate cancer, ovarian cancer, pancreatic cancer, cervical cancer, thyroid cancer, laryngeal cancer, leukemia, acute myeloid cancer. Leukemia, brain tumor, neuroblastoma, retinoblastoma, head and neck cancer, salivary gland cancer, lymphoma, kidney cancer, melanoma, multiple myeloma, brain cancer, osteosarcoma, glioblastoma, IgG opsonization The pharmaceutical composition according to claim 25 , which is any one selected from the group consisting of tumor, lymphoma, neuroma, mesothelioma, and esophageal cancer.

A method for treating or preventing cancer, comprising administering the immune cell according to claim 23 to a subject.