Heterodimeric inactivatable chimeric antigen receptors

Patent metadata

Field	Value
Publication number	US-12528847-B2
Application number	US-201917046760-A
Country	US
Kind code	B2
Filing date	Apr 12, 2019
Priority date	Apr 13, 2018
Publication date	Jan 20, 2026
Grant date	Jan 20, 2026

Abstract

The invention relates to heterodimeric inactivatable chimeric antigen receptors (CARs) and their use for treatment.

First claim

What is claimed is:

1 . A heterodimeric inactivatable chimeric antigen receptor (CAR) comprising:
a) a first polypeptide chain comprising:
i) an extracellular region;
ii) a first transmembrane (TM) region;
iii) a first co-stimulatory endodomain (ED); and
iv) a first member of a dimerization pair; and
b) a second polypeptide chain comprising:
i) optionally, an extracellular region which does not comprise a target-binding region;
ii) a second TM region;
iii) optionally, a second co-stimulatory ED;
iv) a second member of the dimerization pair; and
v) an intracellular signaling ED,
wherein the first and second member of the dimerization pair form a heterodimer, wherein the heterodimer formed by the first and second member of the dimerization pair is disrupted in the presence of an inhibitory molecule resulting in inhibition of CAR-mediated signaling, and wherein (1) the first member of the dimerization pair comprises a sequence selected from SEQ ID NO: 5, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 24 comprising residues 35-50 of SEQ ID NO: 22 in place of residues 51-91 of SEQ ID NO: 24, and SEQ ID NO: 30, and the second member of the dimerization pair comprises a sequence IAXXLXXIGXXF (SEQ ID NO: 179), wherein X is any natural amino acid and L is optionally substituted with A, or (2) the first member of the dimerization pair comprises a sequence IAXXLXXIGXXF (SEQ ID NO: 179), wherein X is any natural amino acid and L is optionally substituted with A, and the second member of the dimerization pair comprises a sequence selected from SEQ ID NO: 5, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 24 comprising residues 35-50 of SEQ ID NO: 22 in place of residues 51-91 of SEQ ID NO: 24, and SEQ ID NO: 30, and the inhibitory molecule is venetoclax, A-1331852, or A-1155463.

2 . The CAR of claim 1 , wherein the second polypeptide chain comprises an extracellular region which does not comprise a target-binding region.

3 . The CAR of claim 1 , wherein the first polypeptide chain does not comprise an intracellular signaling ED.

4 . The CAR of claim 1 , comprising:
a) a first polypeptide chain consisting essentially of in the direction from the N terminus to the C terminus:
i) an extracellular target-binding region;
ii) a first linker region;
iii) a first transmembrane (TM) region;
iv) a first co-stimulatory endodomain (ED); and
v) a first member of a dimerization pair; and
b) a second polypeptide chain consisting essentially of in the direction from the N terminus to the C terminus:
i) an extracellular region which does not comprise a target-binding region;
ii) a second linker region;
iii) a second TM region;
iv) a second co-stimulatory ED;
v) a second member of the dimerization pair; and
vi) an intracellular signaling ED,
wherein the first polypeptide chain does not comprise an intracellular signaling ED.

5 . The CAR of claim 1 , wherein the extracellular region of the first polypeptide chain comprises an antigen-binding polypeptide, a receptor, or a natural ligand for a target cell antigen or receptor.

6 . The CAR of claim 5 , wherein the antigen recognized by the antigen-binding polypeptide is CD19, prostate-specific membrane antigen (PSMA), or mesothelin.

7 . The CAR of claim 5 , wherein the natural ligand for a target cell antigen or receptor is an NKG2D ectodomain.

8 . The CAR of claim 1 , wherein the extracellular region of the first polypeptide chain comprises a T-cell receptor (TCR)-based recognition domain.

9 . The CAR of claim 1 , wherein
a) the first and/or second transmembrane (TM) region is derived from CD8, CD8α, CD4, CD3-zeta, CD3-epsilon, CD28, CD45, CD4, CD5, CD7, CD9, CD16, CD22, CD33, CD37, CD40, CD64, CD80, CD86, CD134 (OX-40), CD137, CD154, DAP10, or DAP12; b) the first and/or second co-stimulatory ED is derived from 4-1BB (CD137), CD28, ICOS, CD134 (OX-40), BTLA, CD27, CD30, GITR, CD226, or HVEM; c) the intracellular signaling ED of the second polypeptide chain is derived from DAP10, DAP12, Fc epsilon receptor I gamma chain (FCER1G), FcR beta CD3-delta, CD3-epsilon, CD3-gamma, CD3-zeta, CD226, CD66d, CD79A, or CD79B; and/or d) the first and/or second polypeptide chain further comprises one or more polypeptide sequences, wherein said one or more polypeptide sequences are selected from one or more additional co-stimulatory EDs, signal sequences, separation sequences, epitope tags, and polypeptides that produce a detectable signal.

10 . The CAR of claim 1 , wherein
i) the first member of the dimerization pair comprises the sequence of SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, or SEQ ID NO: 19; ii) the second member of the dimerization pair comprises the sequence of SEQ ID NO: 5, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, or SEQ ID NO: 30; iii) the extracellular region of the first polypeptide chain comprises the sequence of SEQ ID NO: 6, or SEQ ID NO: 49; iv) the intracellular signaling ED of the second polypeptide chain comprises the sequence of SEQ ID NO: 7; v) the first and/or second linker region comprises the sequence of SEQ ID NO: 9; vi) the first and/or second TM region comprises the sequence of SEQ ID NO: 10; and/or vii) the first and/or second co-stimulatory ED comprises the sequence or SEQ ID NO: 11.

11 . The CAR of claim 1 , wherein:
a) the first polypeptide chain comprises the sequence of SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO: 111, or SEQ ID NO: 112; and/or b) the second polypeptide chain comprises the sequence of SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, or SEQ ID NO: 117.

12 . The CAR of claim 1 comprising:
a) the first polypeptide chain comprising the sequence of SEQ ID NO: 109 and the second polypeptide chain comprising the sequence of SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, or SEQ ID NO: 117;
b) the first polypeptide chain comprising the sequence of SEQ ID NO: 110 and the second polypeptide chain comprising the sequence of SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, or SEQ ID NO: 117;
c) the first polypeptide chain comprising the sequence of SEQ ID NO: 111 and the second polypeptide chain comprising the sequence of SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, or SEQ ID NO: 117; or
d) the first polypeptide chain comprising the sequence of SEQ ID NO: 112 and the second polypeptide chain comprising the sequence of SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, or SEQ ID NO: 117.

13 . A nucleic acid molecule comprising a nucleotide sequence encoding the first polypeptide chain of the heterodimeric inactivatable CAR of claim 1 , the second polypeptide chain of the heterodimeric inactivatable CAR of claim 1 , or the first and second polypeptide chains of the heterodimeric inactivatable CAR of claim 1 .

14 . A recombinant vector comprising the nucleic acid molecule of claim 13 .

15 . An isolated, genetically modified host cell comprising the heterodimeric inactivatable CAR of claim 1 .

16 . The host cell of claim 15 , which is selected from a cytotoxic cell, a T cell, a stem cell, a progenitor cell, a cell derived from a stem cell, and a cell derived from a progenitor cell.

17 . A pharmaceutical composition comprising the host cell of claim 15 and a pharmaceutically acceptable carrier and/or excipient.

18 . A method for producing a host cell comprising the heterodimeric inactivatable CAR of claim 1 , said method comprising genetically modifying said cell with (i) a nucleic acid molecule comprising a nucleotid

Assignees

• Ludwig Inst For Cancer Res Ltd
• Ecole Polytechnique Fed Lausanne Epfl

Inventors

• Coukos George
• Irving Melita
• Correia Bruno
• Gainza-Cirauqui Pablo
• Giordano Attianese Greta Maria Paola

Classifications

• A61K40/4276

  Prostate specific membrane antigen [PSMA] · CPC title

• A61K40/4211

  CD19 or B4 · CPC title

• A61K40/31

  Chimeric antigen receptors [CAR] · CPC title

• A61K40/11

  T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells · CPC title

• A61K2239/58

  Prostate · CPC title