Chimeric antigen receptor

Patent metadata

Field	Value
Publication number	US-2019352409-A1
Application number	US-201716348319-A
Country	US
Kind code	A1
Filing date	Nov 10, 2017
Priority date	Nov 11, 2016
Publication date	Nov 21, 2019
Grant date	—

Abstract

The present invention provides a cell comprising first and second chimeric antigen receptors (CARs), which bind to different antigens, wherein the first CAR binds to Transmembrane activator and calcium modulator and cyclophilin ligand (CAML) interactor (TACI). The present invention also provides a cell comprising a tan CAR comprising first and second antigen-binding domains which bind to different antigens, wherein the first antigen binding domain binds the antigen Transmembrane activator and calcium modulator and cyclophilin ligand (CAML) interactor (TACI). The present invention further provides corresponding nucleic acid sequences and/or constructs, kits and vectors comprising said nucleic acid sequences and/or constructs, molecules and methods for making such cells. The cells may be used in cellular immunotherapy approaches for treating diseases such as multiple myeloma.

First claim

1 . A cell comprising first and second chimeric antigen receptors (CARs), which bind to different antigens, wherein the first CAR binds to Transmembrane activator and calcium modulator and cyclophilin ligand interactor (TACI).

2 . A cell according to claim 1 where the second CAR binds BCMA.

3 . A cell comprising a tanCAR comprising first and second antigen-binding domains which bind to different antigens, wherein the first antigen binding domain binds the antigen Transmembrane activator and calcium modulator and cyclophilin ligand interactor (TACI).

4 . A cell according to claim 3 where the second antigen binding domain binds to BCMA.

5 . A nucleic acid sequence encoding a tanCAR as defined in claim 3 or 4 .

6 . A nucleic acid sequence according to claim 5 , which has the following structure:
AgB1-linker-AgB2-spacer-TM-endo in which AgB1 is a nucleic acid sequence encoding the first antigen-binding domain of the tanCAR; linker is a nucleic acid sequence encoding a linker of the tanCAR; AgB2 is a nucleic acid sequence encoding the second antigen binding domain of the tanCAR; spacer is a nucleic acid sequence encoding a spacer of the tanCAR; TM is a nucleic acid sequence encoding a transmembrane domain of the tanCAR; endo is a nucleic acid sequence encoding an endodomain of the tanCAR; which nucleic acid sequence, when expressed in a cell, encodes a polypeptide expressing the first and second antigen binding domains in tandem at the cell surface.

7 . A nucleic acid sequence according to claim 6 , wherein linker encodes a sequence comprising a Gly-Ser flexible linker.

8 . A nucleic acid sequence according to any of claims 5 to 7 wherein alternative codons are used in regions of sequence encoding the same or similar amino acid sequences, in order to avoid homologous recombination.

9 . A nucleic acid construct which encodes a first CAR as defined in claim 1 or 2 ; and a second CAR as defined in claim 1 or 2 .

10 . A nucleic acid construct according to claim 9 , which has the following structure:
AgB1-spacer1-TM1-endo1-coexpr-AgB2-spacer2-TM2-endo2 in which AgB1 is a nucleic acid sequence encoding the antigen-binding domain of the first CAR; spacer1 is a nucleic acid sequence encoding a spacer of the first CAR; TM1 is a nucleic acid sequence encoding a transmembrane domain of the first CAR; endo1 is a nucleic acid sequence encoding an endodomain of the first CAR; coexpr is a nucleic acid sequence enabling co-expression of the first and second CARs AgB2 is a nucleic acid sequence encoding the antigen-binding domain of the second CAR; spacer2 is a nucleic acid sequence encoding the spacer of the second CAR; TM2 is a nucleic acid sequence encoding the transmembrane domain of the second CAR; endo2 is a nucleic acid sequence encoding the endodomain of the second CAR; which nucleic acid construct, when expressed in a cell, encodes a polypeptide which is cleaved at the cleavage site such that the first and second CARs are co-expressed at the cell surface.

11 . A nucleic acid construct according to claim 10 , wherein coexpr encodes a sequence comprising a self-cleaving peptide.

12 . A nucleic acid construct according to any of claims 9 to 11 , wherein alternative codons are used in regions of sequence encoding the same or similar amino acid sequences, in order to avoid homologous recombination.

13 . A vector comprising a nucleic acid sequence according to any of claims 5 to 8 , or a nucleic acid construct according to any of claims 9 to 12 .

14 . A retroviral vector or a lentiviral vector or a transposon according to claim 13 .

15 . A kit which comprises
(i) a first nucleic acid sequence encoding a first CAR as defined in claim 1 or 2 , which nucleic acid sequence has the following structure: AgB1-spacer1-TM1-endo1 in which AgB1 is a nucleic acid sequence encoding the antigen-binding domain of the first CAR; spacer 1 is a nucleic acid sequence encoding the spacer of the first CAR; TM1 is a nucleic acid sequence encoding the transmembrane domain of the first CAR; endo 1 is a nucleic acid sequence encoding the endodomain of the first CAR; and (ii) a second nucleic acid sequence encoding a second CAR as defined in claim 1 or 2 , which nucleic acid sequence has the following structure: AgB2-spacer2-TM2-endo2 in which AgB2 is a nucleic acid sequence encoding the antigen-binding domain of the second CAR; spacer 2 is a nucleic acid sequence encoding the spacer of the second CAR; TM2 is a nucleic acid sequence encoding the transmembrane domain of the second CAR; endo 2 is a nucleic acid sequence encoding the endodomain of the second CAR.

16 . A kit comprising: a first vector which comprises the first nucleic acid sequence as defined in claim 15 ; and a second vector which comprises the second nucleic acid sequence as defined in claim 15 .

17 . A kit according to claim 16 , wherein the vectors are integrating viral vectors or transposons.

18 . A method for making a cell according to any of claims 1 to 4 , which comprises the step of introducing: a nucleic acid sequence according to any of claims 5 to 8 ; a nucleic acid construct according to any of claims 10 to 12 ; a vector according to claim 13 or 14 or a kit of sequences or vectors according to any of claims 15 to 17 , into a cell.

19 . A method according to claim 18 , wherein the cell is from a sample isolated from a subject.

20 . A pharmaceutical composition comprising a plurality of cells according to any of claims 1 to 4 .

21 . A pharmaceutical composition comprising:
(i) a first cell population expressing a first CAR; and (ii) a second cell population expressing a second CAR
wherein the first and second CARs bind different antigens and the first CAR binds to Transmembrane activator and calcium modulator and cyclophilin ligand interactor (TACI).

22 . A method for treating and/or preventing a disease, which comprises the step of administering a pharmaceutical composition according to claim 20 or 21 to a subject.

23 . A method for treating and/or preventing a disease, which comprises the following steps:
(i) administration of a first cell population which expresses a first CAR (ii) administration of a second cell population which expresses a second CAR wherein the first and second CARs bind different antigens and wherein the first CAR binds to Transmembrane activator and calcium modulator and cyclophilin ligand interactor (TALI).

24 . A method according to claim 22 or 23 , which comprises the following steps:
(i) isolation of a cell-containing sample from a subject;
(ii) transduction or transfection of the cells with: a nucleic acid sequence according to any of claims 5 to 8 ; a nucleic acid construct according to any of claims 10 to 12 ; a vector according to claim 13 or 14 or a kit of sequences or vectors according to any of claims 15 to 17 ; and
(iii) administering the cells from (ii) to the subject.

25 . A kit for use in a method according to claim 23 , which comprises:
(i) a first cell population which expresses a first CAR (ii) a second cell population which expresses a second CAR wherein the first and second CARs bind different antigens and wherein the first CAR binds to Transmembrane activator and calcium modulator and cyclophilin ligand interactor (TALI).

26 . A method according to a

Assignees

• Autolus Ltd

Inventors

• Onuoha Shimobi
• Peddareddigari Vijay
• Stanczuk Lukas
• Ferrari Mathieu
• Ma Biao

Classifications

• A61P35/00

  Antineoplastic agents · CPC title

• C07K16/2878 Primary

  against the NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95 · CPC title

• C07K2317/622

  Single chain antibody (scFv) · CPC title

• C07K2319/32

  fusions with soluble part of a cell surface receptor, "decoy receptors" · CPC title

• C07K2319/03

  containing a transmembrane segment · CPC title