Tunable Chimeric Antigen Receptors

Patent metadata

Field	Value
Publication number	US-2019330337-A1
Application number	US-201716310076-A
Country	US
Kind code	A1
Filing date	Jun 15, 2017
Priority date	Jun 16, 2016
Publication date	Oct 31, 2019
Grant date	—

Abstract

The present invention provides a cell which co-expresses a first chimeric antigen receptor (CAR) and second CAR at the cell surface, each CAR comprising an antigen-binding domain, a transmembrane domain and an intracellular domain wherein the antigen-binding domain of the first CAR binds to CD19 and the antigen-binding domain of the second CAR binds to CD22; and wherein the first and/or second CAR is a tunable CAR having an intracellular domain comprising a heterodimenzation domain, which intracellular domain is capable of binding a separate intracellular signalling molecule which comprises a reciprocal heterodimenzation domain and a signalling domain.

First claim

1 . A cell which co-expresses a first chimeric antigen receptor (CAR) and second CAR at the cell surface, each CAR comprising an antigen-binding domain, a transmembrane domain and an intracellular domain
wherein the antigen-binding domain of the first CAR binds to CD19 and the antigen-binding domain of the second CAR binds to CD22; and wherein the first and/or second CAR is a tunable CAR having an intracellular domain comprising a heterodimerization domain, which intracellular domain is capable of binding a separate intracellular signalling molecule which comprises a reciprocal heterodimerization domain and a signalling domain.

2 . A cell according to claim 1 , wherein binding of the first and second/or CAR to the intracellular signalling molecule is disrupted by the presence of an agent, such that in the absence of the agent the first and/or second CAR heterodimerize(s) with the intracellular signalling molecule and binding of the antigen binding domain to antigen results in signalling through the signalling domain; whereas in the presence of the agent, the first and/or second CAR do/does not heterodimerize with the intracellular signalling molecule and binding of the antigen binding domain to antigen does not result in signalling through the signalling domain.

3 . A cell according to claim 1 , wherein the first CAR, which binds to CD19, is a tunable CAR, having an intracellular domain which comprises a heterodimerization domain which binds a heterodimerization domain of an intracellular signalling molecule; and the second CAR, which binds to CD22, is a classical CAR, having an intracellular domain which comprises a signalling domain.

4 - 17 . (canceled)

18 . A nucleic acid construct encoding a first chimeric antigen receptor (CAR) and second CAR, each CAR comprising an antigen-binding domain, a transmembrane domain and an intracellular domain
wherein the antigen-binding domain of the first CAR binds to CD19 and the antigen-binding domain of the second CAR binds to CD22; and wherein the first and/or second CAR is a tunable CAR having an intracellular domain comprising a heterodimerization domain, which intracellular domain is capable of binding a separate intracellular signalling molecule which comprises a reciprocal heterodimerization domain and a signalling domain.

19 . A nucleic acid construct according to claim 18 , which has one of the following structures:
a) AgB1-spacer1-TM1-HD1-coexpr-AgB2-spacer2-TM2-endo2; b) AgB1-spacer1-TM1-endo1-coexpr-AgB2-spacer2-TM2-HD2; c) AgB2-spacer2-TM2-HD2-coexpr-AgB1-spacer1-TM1-endo1; d) AgB2-spacer2-TM2-endo2-coexpr-AgB1-spacer1-TM1-HD1; e) AgB1-spacer1-TM1-HD1-coexpr-AgB2-spacer2-TM1-HD1; f) AgB2-spacer2-TM2-HD2-coexpr-AgB1-spacer1-TM1-HD1 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 HD1 is a nucleic acid sequence encoding a heterodimerisation domain of the first CAR Endo1 is a nucleic acid sequence encoding an intracellular domain which comprises a signalling domain of the first CAR; coexpr is a nucleic acid sequence enabling co-expression of both CARs 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 a nucleic acid sequence encoding the transmembrane domain of the second CAR; HD2 is a nucleic acid sequence encoding a heterodimerisation domain of the second CAR Endo2 is a nucleic acid sequence encoding an intracellular domain which comprises a signalling domain of the second CAR.

20 - 21 . (canceled)

22 . A nucleic acid construct according to claim 18 , which also comprises a nucleic acid sequence encoding an intracellular signalling molecule which comprises a heterodimerization domain reciprocal to the heterodimerization domain on the tunable CAR, and a signalling domain.

23 . A kit which comprises
a first nucleic acid sequence encoding a first chimeric antigen receptor (CAR) and second nucleic acid sequence encoding a second CAR, each CAR comprising an antigen-binding domain, a transmembrane domain and an intracellular domain wherein the antigen-binding domain of the first CAR binds to CD19 and the antigen-binding domain of the second CAR binds to CD22; and wherein the first and/or second CAR is a tunable CAR having an intracellular domain comprising a heterodimerization domain, which intracellular domain is capable of binding a separate intracellular signalling molecule which comprises a reciprocal heterodimerization domain and a signalling domain wherein
(i) the first nucleic acid sequence has the following structures:
AgB1-spacer1-TM1-HD1 or 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; HD1 is a nucleic acid sequence encoding a heterodimerisation domain of the first CAR; and Endo1 is a nucleic acid sequence encoding an intracellular domain which comprises a signalling domain of the first CAR and
(ii) the second nucleic acid sequence has the following structure:
AgB2-spacer2-TM2-HD2 or 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; HD2 is a nucleic acid sequence encoding a heterodimerisation domain of the second CAR; and Endo2 is a nucleic acid sequence encoding an intracellular domain which comprises a signalling domain of the second CAR.

24 . A kit according to claim 23 , which also comprises (iii) a third nucleic acid sequence encoding an intracellular signalling molecule which comprises a heterodimerization domain reciprocal to the heterodimerization domain on the tunable CAR, and a signalling domain.

25 . A kit comprising: a first vector which comprises a first nucleic acid sequence encoding a first chimeric antigen receptor (CAR); and a second vector which comprises a second nucleic acid sequence encoding a second CAR, each CAR comprising an antigen-binding domain, a transmembrane domain and an intracellular domain
wherein the antigen-binding domain of the first CAR binds to CD19 and the antigen-binding domain of the second CAR binds to CD22; and wherein the first and/or second CAR is a tunable CAR having an intracellular domain comprising a heterodimerization domain, which intracellular domain is capable of binding a separate intracellular signalling molecule which comprises a reciprocal heterodimerization domain and a signalling domain.

26 . A vector comprising a nucleic acid construct according to claim 18 .

27 . (canceled)

28 . A method for making a cell according to claim 1 , which comprises the step of introducing: a nucleic acid construct according to claim 18 into a cell.

29 . (canceled)

30 . A pharmaceutical composition comprising a plurality of cells according to claim 1 .

31 . A method for treating and/or preventing a disease, which comprises the step of administering a pharmaceutical composition according to claim 30 to a subject.

32 . A method according to claim 31 , which comprises the following steps:
(i) isolation of a cell-containing

Assignees

• Autolus Ltd

Inventors

• Pulé Martin
• Thomas Simon
• Cordoba Shaun
• Kokalaki Evangelia

Classifications

• C07K2317/524

  CH2 domain · CPC title

• A61K2039/507

  Comprising a combination of two or more separate antibodies · CPC title

• C07K2319/03

  containing a transmembrane segment · CPC title

• A61K39/39558 Primary

  against tumor tissues, cells, antigens · CPC title

• C07K14/70578

  NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95 (NGF-receptor C07K14/71, TNF-receptor C07K14/7151) · CPC title