REGULATABLE CHIMERIC ANTIGEN RECEPTOR

• US 20170081411A1
• Filed: 03/13/2015
• Published: 03/23/2017
• Est. Priority Date: 03/15/2014
• Status: Abandoned Application

• Current Assignee: Novartis Ag, Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Original Assignee: Novartis Ag, Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Inventors: Engels, Boris, Loew, Andreas, Milone, Michael C., Zhou, Li
• Application Number:
  US15/126,037
• Publication Number:
  US 20170081411A1
• Time in Patent Office: Days
• CPC Class Codes: A61K 2039/5156 expressing foreign proteins
• CPC Class Codes: A61K 2039/5158 Antigen-pulsed cells, e.g. T-cells
• CPC Class Codes: A61K 2039/55561 CpG containing adjuvants; Oligonucleotide containing adjuvants
• CPC Class Codes: A61K 2239/23 On/off switch
• CPC Class Codes: A61K 2239/24 Dimerizable CARs; CARs with adapter
• CPC Class Codes: A61K 2239/29 Multispecific CARs
• CPC Class Codes: A61K 2239/48 Blood cells, e.g. leukemia or lymphoma
• CPC Class Codes: A61K 2239/59 Reproductive system, e.g. uterus, ovaries, cervix or testes
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 38/177 Receptors; Cell surface antigens; Cell surface determinants
• CPC Class Codes: A61K 38/1774 Immunoglobulin superfamily (e.g. CD2, CD4, CD8, ICAM molecules, B7 molecules, Fc-receptors, MHC-molecules)
• CPC Class Codes: A61K 39/0011 Cancer antigens
• CPC Class Codes: A61K 39/3955 against proteinaceous materials, e.g. enzymes, hormones, lymphokines
• CPC Class Codes: A61K 39/4611 T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
• CPC Class Codes: A61K 39/4613 Natural-killer cells [NK or NK-T]
• CPC Class Codes: A61K 39/4631 Chimeric Antigen Receptors [CAR]
• CPC Class Codes: A61K 39/4636 Immune checkpoint inhibitors
• CPC Class Codes: A61K 39/464412 CD19 or B4
• CPC Class Codes: A61K 39/464468 Mesothelin [MSLN]
• CPC Class Codes: C07K 14/705 Receptors; Cell surface antigens; Cell surface determinants tumour specific antigens C07K14/4748
• CPC Class Codes: C07K 14/7051 : T-cell receptor (TcR)-CD3 complex
• CPC Class Codes: C07K 14/70517 : CD8
• CPC Class Codes: C07K 14/70575 : NGF/TNF-superfamily, e.g. CD70, CD95L, CD153, CD154 NGF C07K14/48, TNF C07K14/525
• CPC Class Codes: C07K 16/00 : Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies antibodies with enzymatic activity, e.g. abzymes C12N9/0002
• CPC Class Codes: C07K 16/28 : against receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: C07K 16/2803 : against the immunoglobulin superfamily
• CPC Class Codes: C07K 16/2863 : against receptors for growth factors, growth regulators
• CPC Class Codes: C07K 2317/22 : from camelids, e.g. camel, llama or dromedary
• CPC Class Codes: C07K 2317/53 : Hinge
• CPC Class Codes: C07K 2317/622 : Single chain antibody (scFv)
• CPC Class Codes: C07K 2319/02 : containing a signal sequence
• CPC Class Codes: C07K 2319/03 : containing a transmembrane segment
• CPC Class Codes: C07K 2319/10 : containing a tag for extracellular membrane crossing, e.g. TAT or VP22
• CPC Class Codes: C07K 2319/30 : Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
• CPC Class Codes: C07K 2319/33 : fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies
• CPC Class Codes: C07K 2319/70 : containing domain for protein-protein interaction
• CPC Class Codes: C07K 2319/73 : containing coiled-coiled motif (leucine zippers)
• CPC Class Codes: C12N 15/62 : DNA sequences coding for fusion proteins

Abstract

Provided are compositions and methods relating to regulatable chimeric antigen receptors (RCARs), natural killer cell receptor CARs (NKR-CARs), and regulatable NKR-CARs (RNKR-CARs), where the intracellular signaling or proliferation of the RCAR or RNKR-CAR can be controlled to optimize the use of an RCAR/NKR-CAR- or RNKR-CAR-expressing cell to provide an immune response. Cells can be engineered to express a RNKR-CAR or to express a RCAR and a NKR-CAR (e.g., inhibitory NKR-CAR). For example, a RCAR or RNKR-CAR can comprise a dimerization switch that, upon the presence of a dimerization molecule, can couple an intracellular signaling domain to an extracellular recognition element, e.g., an antigen binding domain, an inhibitory counter ligand binding domain, or costimulatory ECD domain. An RCAR or RNKR-CAR can be engineered to include an appropriate antigen binding domain that is specific to a desired antigen target and used in the treatment of a disease.

Claims

1. A regulatable natural killer receptor CAR (RNKR-CAR), e.g., an isolated NKR-CAR, wherein the RNKR-CAR comprises:

c) an antigen binding member, comprising

a binding domain element,

a transmembrane domain,

a first switch domain, and

optionally, a NKR cytoplasmic domain, e.g., selected from Table 24,

wherein the binding domain element comprises an antigen binding domain, an inhibitory extracellular domain, e.g., selected from Table 4, or a costimulatory extracellular domain, e.g., selected from Table 5; and

d) an intracellular signaling member comprising

a second switch domain,

a NKR cytoplasmic domain e.g., selected from Table 24, or an intracellular signaling domain, e.g., a primary signaling domain, e.g., selected from Table 1, e.g., a DAP12 signaling domain, or a CD3zeta signaling domain, and

optionally, a transmembrane domain or a membrane tether.

2. The RNKR-CAR of claim 1, wherein:

a) the RNKR-CAR comprises an NKR cytoplasmic domain or an NKR transmembrane domain, and the NKR cytoplasmic domain, if present, is other than a FcR gamma (FCER1G), CD27, NKG2C, SLAMF7, NKP80 (KLRF1), CD160 (BY55), DNAM1 (CD226), SLAMF4 (CD244, 2B4), CD84, CD96 (Tactile), CEACAM1, CRTAM, Ly9 (CD229), PSGL1, CD100 (SEMA4D), SLAMF6 (NTB-A, Ly108), SLAM (SLAMF1, CD150, IPO-3), BLAME (SLAMF8), NKp44, NKp30, and/or NKp46 cytoplasmic domain;

b) the RNKR-CAR comprises an NKR cytoplasmic domain and a primary signaling domain from an NK cell adaptor molecule, e.g., DAP12;

c) the RNKR-CAR comprises an NKR transmembrane domain and a primary signaling domain from an NK cell adaptor molecule, e.g., DAP12;

d) the RNKR-CAR comprises an NKR cytoplasmic domain (other than a FcR gamma (FCER1G), CD27, NKG2C, SLAMF7, NKP80 (KLRF1), CD160 (BY55), DNAM1 (CD226), SLAMF4 (CD244, 2B4), CD84, CD96 (Tactile), CEACAM1, CRTAM, Ly9 (CD229), PSGL1, CD100 (SEMA4D), SLAMF6 (NTB-A, Ly108), SLAM (SLAMF1, CD150, IPO-3), BLAME (SLAMF8), NKp44, NKp30, and/or NKp46 cytoplasmic domain) and a primary signaling domain from a T cell molecule, e.g., CD3zeta;

e) the RNKR-CAR comprises a mutated NKR transmembrane domain;

or

f) the RNKR-CAR comprises a transmembrane domain other than a NKR transmembrane domain, e.g., comprises a transmembrane domain from a T cell molecule, e.g., CD8alpha or CD3zeta.

3. The RNKR-CAR of claim 1 or 2, wherein the RNKR-CAR comprises

a regulatable killer immunoglobulin receptor-CAR (RKIR-CAR), e.g., an RactKIR-CAR or RinhKIR-CAR;

a RNCR-CAR, e.g., an RactRNCR-CAR;

a RFcR-CAR, e.g., an RactCD16-CAR or an RactCD64-CAR;

a RLy49-CAR, e.g., an RactLy49-CAR or an RinhLy49-CAR;

or

a RSLAMF-CAR, RinhSLAMF-CAR.

4. The RNKR-CAR of any of claims 1-3, wherein the first and second switch domains comprise a dimerization switch.

5. A nucleic acid encoding a RNKR-CAR of any of the previous claims.

6. A vector system, e.g., one or more vectors, comprising a nucleic acid of claim 5.

7. A cell comprising a RNKR-CAR of any of claims 1-4, a nucleic acid encoding a RNKR-CAR of claim 5, or the vector system of claim 6.

8. A method of making a cell of claim 7, comprising introducing a nucleic acid encoding a RNKR-CAR of claim 5, or the vector system of claim 6 into said cell.

9. A method of treating a subject with a disease associated with a tumor antigen comprising administering to the subject an effective amount of a RNKR-CAR cell of claim 7.

10. A method of providing a RNKR-CAR cell of claim 7 comprising:

providing an immune effector cell to a recipient entity; and

receiving from said entity, a RNKR-CAR cell derived from said immune effector cell, or a daughter cell thereof, wherein the RNKR-CAR comprises an RNKR-CAR of any of claims 1-4, or a nucleic acid or vector encoding the RNKR-CAR.

11. A method of providing an RNKR-CAR cell comprising:

receiving from an entity an immune effector cell from a human;

inserting a nucleic acid encoding an RNKR-CAR of any of claims 1-4 into said immune effector cell, or a daughter cell thereof, to form an RNKR-CAR cell; and

, optionally, providing said RNKR-CAR cell to said entity.

12. A RCAR/NKR-CAR cell comprising:

A) a regulatable CAR (RCAR) and a NKR-CAR;

B) a nucleic acid encoding a RCAR and a NKR-CAR;

or

C) a vector system comprising a nucleic acid encoding a RCAR and a NKR-CAR, wherein the RCAR comprises;

a) an intracellular signaling member comprising;

an intracellular signaling domain, e.g., a primary intracellular signaling domain, and

a first switch domain;

b) an antigen binding member comprising;

an antigen binding domain,

a second switch domain; and

optionally, one or a plurality, of co-stimulatory signaling domain, and

c) optionally, a transmembrane domain; and

wherein the NKR-CAR comprises;

a) an antigen binding domain,

b) a transmembrane domain, e.g., an NKR transmembrane domain, and

c) a cytoplasmic domain, e.g., an NKR cytoplasmic domain.

13. The cell of claim 12, wherein the NKR-CAR comprises an inhibitory NKR-CAR (inhNKR-CAR).

14. The cell of claim 12 or 13, wherein the antigen binding domain of the RCAR and the antigen binding domain of the NKR-CAR, e.g., the inhNKR-CAR, target different antigens.

17. A method of treating a subject with a disease associated with a tumor antigen comprising administering to the subject an effective amount of a RCAR/NKR-CAR cell of any of claims 12-14.

18. A method of providing a RCAR/NKR-CAR cell of any of claims 12-14 comprising:

providing an immune effector cell to a recipient entity; and

receiving from said entity, a RCAR/NKR-CAR cell derived from said immune effector cell, or a daughter cell thereof, wherein the RCAR/NKR-CAR comprises;

a) a regulatable CAR (RCAR) and a NKR-CAR;

b) a nucleic acid encoding a RCAR and a NKR-CAR;

c) a first nucleic acid encoding a RCAR and a second nucleic acid encoding a NKR-CAR;

or

d) a vector system comprising a nucleic acid encoding a RCAR and a NKR-CAR or comprising a first nucleic acid encoding a RCAR and a second nucleic acid encoding a NKR-CAR.

15. A nucleic acid comprising

(iii) a sequence encoding a RCAR and

(iv) a sequence encoding a NKR-CAR,

wherein the RCAR comprises:

A) an intracellular signaling member comprising;

an intracellular signaling domain, e.g., a primary intracellular signaling domain, and

a first switch domain;

B) an antigen binding member comprising;

an antigen binding domain,

a second switch domain; and

optionally,

C) a transmembrane domain, and

wherein the NKR-CAR comprises;

a) an antigen binding domain,

b) a transmembrane domain, e.g., an NKR transmembrane domain, and

c) a cytoplasmic domain, e.g., an NKR cytoplasmic domain.

16. A vector system comprising a nucleic acid of claim 15.

19. A method of providing an RCAR/NKR-CAR cell comprising:

receiving from an entity an immune effector cell from a human;

inserting into said immune effector cell or a daughter cell thereof one of the following;

a) a nucleic acid encoding a RCAR and a NKR-CAR, or b) a first nucleic acid encoding a RCAR and a second nucleic acid encoding a NKR-CAR, to form a RCAR/NKR-CAR cell; and

, optionally, providing said RCAR/NKR-CAR cell to said entity.