TREATMENT OF CANCER USING CHIMERIC CD3 RECEPTOR PROTEINS

• US 20180230193A1
• Filed: 08/05/2016
• Published: 08/16/2018
• Est. Priority Date: 08/07/2015
• Status: Active Grant

• 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: Loew, Andreas, Granda, Brian, Ramones, Melissa
• Granted Patent: US 11,667,691 B2
• Time in Patent Office: Days
• CPC Class Codes: C07K 14/7051 T-cell receptor (TcR)-CD3 complex
• CPC Class Codes: C07K 16/2803 against the immunoglobulin superfamily
• CPC Class Codes: C07K 16/30 from tumour cells
• CPC Class Codes: C07K 19/00 Hybrid peptides , i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes
• CPC Class Codes: C07K 2317/622 Single chain antibody (scFv)
• CPC Class Codes: C07K 2317/73 Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
• CPC Class Codes: C07K 2319/03 containing a transmembrane segment
• CPC Class Codes: C07K 2319/70 containing domain for protein-protein interaction

Abstract

The present disclosure features the use of chimeric CD3 proteins to modulate T cell Receptor (TCR) signaling. Specifically, the disclosure is based, in part, on the discovery that chimeric CD3 proteins (e.g., CD3delta, CD3gamma, and CD3epsilon) having all or most of their extracellular domain fused to an antigen binding domain can activate the TCR in the presence of a cognate antigen. The disclosure is further based on the observation that the above chimeric proteins can be potentiated through the inclusion of a co-stimulatory domain in the intracellular portion of the chimeric molecule. Thus, the preferred elements of the engineered signaling complexes of the disclosure include an antigen binding domain, an extracellular domain derived from one of the above CD3 proteins, and an intracellular co-stimulatory domain.

Claims

1. A chimeric membrane protein comprising a CD3 gamma, delta, or epsilon domain and an intracellular co-stimulatory domain, wherein said CD3 domain comprises an extracellular domain derived from the extracellular domain of CD3 gamma, delta, or epsilon and said intracellular co-stimulatory domain is not derived from CD3 gamma, delta, or epsilon.

4. The chimeric membrane protein of claim 1, further comprising an antigen binding domain.

5. The chimeric membrane protein of claim 1, wherein said CD3 gamma, delta, or epsilon domain comprises the entire CD3 gamma, delta, or epsilon extracellular domain or the entire CD3 gamma, delta, or epsilon protein.

7. The chimeric membrane protein of claim 1, wherein said chimeric protein does not comprise any intracellular domain derived from CD3 gamma, delta, or epsilon.

9. The chimeric membrane protein of claim 4, wherein the antigen binding domain is located N-terminal to said CD3 gamma, delta, or epsilon domain.

43. The chimeric membrane protein of claim 4, wherein one or more of said intracellular co-stimulatory domains is a functional signaling domain of a protein selected from the group consisting of:

an MHC class I molecule, TNF receptor proteins, Immunoglobulin-like proteins, cytokine receptors, integrins, signaling lymphocytic activation molecules (SLAM proteins), activating NK cell receptors, BTLA, a Toll ligand receptor, OX40, CD2, CD7, CD27, CD28, CD30, CD40, CDS, ICAM-1, LFA-1 (CD11a/CD18), 4-1BB (CD137), B7-H3, CDS, ICAM-1, ICOS (CD278), GITR, BAFFR, LIGHT, HVEM (LIGHTR), KIRDS2, SLAMF7, NKp80 (KLRF1), NKp44, NKp30, NKp46, CD19, CD4, CD8alpha, CD8beta, IL2R beta, IL2R gamma, IL7R alpha, ITGA4, VLA1, CD49a, ITGA4, IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD, CD11d, ITGAE, CD103, ITGAL, CD11a, LFA-1, ITGAM, CD11b, ITGAX, CD11c, ITGB1, CD29, ITGB2, CD18, LFA-1, ITGB7, NKG2D, NKG2C, TNFR2, TRANCE/RANKL, DNAM1 (CD226), SLAMF4 (CD244, 2B4), CD84, CD96 (Tactile), CEACAM1, CRTAM, Ly9 (CD229), CD160 (BY55), PSGL1, CD100 (SEMA4D), CD69, SLAMF6 (NTB-A, Ly108), SLAM (SLAMF1, CD150, IPO-3), BLAME (SLAMF8), SELPLG (CD162), LTBR, LAT, GADS, SLP-76, PAG/Cbp, CD19a, and a ligand that specifically binds with CD83.

52. A cell comprising the chimeric membrane protein of claim 1.

53. A nucleic acid construct encoding the chimeric membrane protein of claim 1.

54. A vector comprising the nucleic acid construct of claim 53, wherein said vector is a lentiviral, adenoviral, or retroviral vector.

57. A method of treating a subject with a proliferative disorder, said method comprising administering a cell comprising the chimeric membrane protein of claim 4.

2. A chimeric membrane protein comprising a CD3 gamma, delta, or epsilon domain and a first intracellular dimerization domain, wherein said CD3 gamma, delta, or epsilon domain comprises an extracellular domain derived from the extracellular domain of CD3 gamma, delta, or epsilon.

3. The chimeric membrane protein of claim 2, wherein said protein further comprises an intracellular co-stimulatory domain.

10. A cell comprising the chimeric membrane protein of claim 2.

11. A nucleic acid construct encoding the chimeric membrane protein of claim 2.

12. The nucleic acid construct of claim 11, wherein said nucleic acid construct is mRNA.

13. A vector comprising the nucleic acid construct of claim 11, wherein said vector is a lentiviral, adenoviral, or retroviral vector.

15. The cell of claim 10, wherein said chimeric membrane protein comprises said CD3 gamma, delta, or epsilon domain and intracellular dimerization domain, and said cell further comprises a second chimeric protein, said second chimeric protein comprising an extracellular antigen binding domain, an intracellular costimulatory domain and a second intracellular dimerization domain.

17. The cell of claim 10, wherein said chimeric membrane protein comprises said CD3 gamma, delta, or epsilon domain and intracellular dimerization domain, and said cell further comprises a second chimeric protein, said second chimeric protein comprising an extracellular antigen binding domain, an intracellular costimulatory domain, and a CD3 gamma, delta, or epsilon binding domain.

20. The cell of claim 17, wherein said CD3 epsilon binding domain is derived from an anti-CD3 gamma, delta, or epsilon antibody.

21. The cell of claim 20, wherein the CD3 epsilon binding domain comprises the sequence of an scFv or Vhh domain, wherein said CD3 gamma, delta, or epsilon binding domain is an intracellular domain.

22. The cell of claim 15, wherein said extracellular antigen-binding domain of said second chimeric protein is heterologous to said intracellular costimulatory signaling domain of said second chimeric protein.

23. The cell of claim 15, wherein said extracellular antigen-binding domain of said second chimeric protein is the extracellular domain of an inhibitory molecule.

24. The cell of claim 15, wherein said extracellular antigen-binding domain of said second chimeric protein is naturally associated with said intracellular costimulatory signaling domain of said second chimeric protein.

25. The cell of claim 15, wherein said extracellular antigen binding-domain of said second chimeric protein comprises the antigen binding domain of an antibody or fragment thereof.

26. The cell of claim 15, wherein said second chimeric protein is expressed as an intracellular protein.

27. The cell of claim 15, wherein said first and second dimerization domains make up a heterodimerization pair and heterodimerize when expressed in said cell.

28. The cell of claim 27, wherein the heterodimerization domain pairs are selected from the group consisting of:

FKBP and FRP, D53 and MDM2, mFos and mJun Coils, VPS36 and VPS28, GyrB-GyrB based switch, a GAI-GID1 based switch, or a Halo-tag/SNAP-tag based switch.

30. The cell of claim 15, wherein said first and second dimerization domain make up a heterodimerization pair and heterodimerize when expressed in said cell only in the presence of a dimerization compound.

31. The cell of claim 28, wherein one of said first and second dimerization domains comprises a rapamycin analog binding sequence having at least 85% identity with FRP and the other dimerization domain comprises a rapamycin analog binding sequence having at least 85% identity with FKBP.

32. The cell of claim 31, wherein one of said first and second dimerization domains comprises a rapamycin analog binding sequence from FKBP.

33. The cell of claim 31, wherein the other of said first and second dimerization domain comprises a rapamycin analog binding sequence from FRP.

36. The cell of claim 15, wherein said first and second chimeric protein both comprise an intracellular co-stimulatory domain derived from the same protein or different endogenous proteins.

38. The cell of claim 15, wherein said cell is selected from an NK cell or T cell.

39. A nucleic acid encoding the first and second chimeric proteins of claim 15.

40. A vector comprising the nucleic acid of claim 39.

41. The vector of claim 40, wherein, upon expression of said first and second chimeric proteins, said proteins are expressed as a single mRNA transcript.

42. The vector of claim 41, wherein the nucleic acid sequences encoding said first and second chimeric proteins are separated by a nucleic acid encoding a self-cleavage site or an internal ribosomal entry site.

44. A method of treating a subject with a proliferative disorder, said method comprising administering the cell of claim 15.

46. The method of claim 44, wherein said cell is a T cell or NK cell and is autologous or allogeneic to said subject.

48. The method of claim 44, wherein said subject is a human.

51. The chimeric membrane protein of claim 3, wherein said protein further comprises an antigen binding domain.

55. The cell of claim 15, wherein the antigen binding domain is chosen from CD19, CD123, CD22, CD30, CD171, CS-1, CLL-1 (CLECL1), CD33, EGFRvIII, GD2, GD3, BCMA, Tn Ag, PSMA, ROR1, FLT3, FAP, TAG72, CD38, CD44v6, CEA, EPCAM, B7H3, KIT, IL-13Ra2, Mesothelin, IL-11Ra, PSCA, VEGFR2, LewisY, CD24, PDGFR-beta, PRSS21, SSEA-4, CD20, Folate receptor alpha, ERBB2 (Her2/neu), MUC1, EGFR, NCAM, Prostase, PAP, ELF2M, Ephrin B2, IGF-I receptor, CAIX, LMP2, gp100, bcr-abl, tyrosinase, EphA2, Fucosyl GM1, sLe, GM3, TGS5, HMWMAA, o-acetyl-GD2, Folate receptor beta, TEM1/CD248, TEM7R, CLDN6, TSHR, GPRC5D, CXORF61, CD97, CD179a, ALK, Polysialic acid, PLAC1, GloboH, NY-BR-1, UPK2, HAVCR1, ADRB3, PANX3, GPR20, LY6K, OR51E2, TARP, WT1, NY-ESO-1, LAGE-1a, legumain, HPV E6,E7, MAGE-A1, MAGE A1, ETV6-AML, sperm protein 17, XAGE1, Tie 2, MAD-CT-1, MAD-CT-2, Fos-related antigen 1, p53, p53 mutant, prostein, survivin and telomerase, PCTA-1/Galectin 8, MelanA/MART1, Ras mutant, hTERT, sarcoma translocation breakpoints, ML-IAP, ERG (TMPRSS2 ETS fusion gene), NA17, PAX3, Androgen receptor, Cyclin B1, MYCN, RhoC, TRP-2, CYP1B1, BORIS, SART3, PAX5, OY-TES1, LCK, AKAP-4, SSX2, RAGE-1, human telomerase reverse transcriptase, RU1, RU2, intestinal carboxyl esterase, mut hsp70-2, CD79a, CD79b, CD72, LAIR1, FCAR, LILRA2, CD300LF, CLEC12A, BST2, EMR2, LY75, GPC3, FCRL5, or IGLL1.

56. The chimeric membrane protein of claim 51, wherein one or more of said intracellular co-stimulatory domains is a functional signaling domain of a protein selected from the group consisting of:

an MHC class I molecule, TNF receptor proteins, Immunoglobulin-like proteins, cytokine receptors, integrins, signaling lymphocytic activation molecules (SLAM proteins), activating NK cell receptors, BTLA, a Toll ligand receptor, OX40, CD2, CD7, CD27, CD28, CD30, CD40, CDS, ICAM-1, LFA-1 (CD11a/CD18), 4-1BB (CD137), B7-H3, CDS, ICAM-1, ICOS (CD278), GITR, BAFFR, LIGHT, HVEM (LIGHTR), KIRDS2, SLAMF7, NKp80 (KLRF1), NKp44, NKp30, NKp46, CD19, CD4, CD8alpha, CD8beta, IL2R beta, IL2R gamma, IL7R alpha, ITGA4, VLA1, CD49a, ITGA4, IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD, CD11d, ITGAE, CD103, ITGAL, CD11a, LFA-1, ITGAM, CD11b, ITGAX, CD11c, ITGB1, CD29, ITGB2, CD18, LFA-1, ITGB7, NKG2D, NKG2C, TNFR2, TRANCE/RANKL, DNAM1 (CD226), SLAMF4 (CD244, 2B4), CD84, CD96 (Tactile), CEACAM1, CRTAM, Ly9 (CD229), CD160 (BY55), PSGL1, CD100 (SEMA4D), CD69, SLAMF6 (NTB-A, Ly108), SLAM (SLAMF1, CD150, IPO-3), BLAME (SLAMF8), SELPLG (CD162), LTBR, LAT, GADS, SLP-76, PAG/Cbp, CD19a, and a ligand that specifically binds with CD83.

60. The cell of claim 17, wherein said first and second dimerization domains make up a heterodimerization pair and heterodimerize when expressed in said cell.

61. The cell of claim 60, wherein the heterodimerization domain pairs are selected from the group consisting of:

FKBP and FRP, p53 and MDM2, mFos and mJun Coils, VPS36 and VPS28, GyrB-GyrB based switch, a GAI-GID1 based switch, or a Halo-tag/SNAP-tag based switch.

62. The cell of claim 17, wherein said first and second dimerization domain make up a heterodimerization pair and heterodimerize when expressed in said cell only in the presence of a dimerization compound.

63. The cell of claim 60, wherein one of said first and second dimerization domains comprises a rapamycin analog binding sequence having at least 85% identity with FRP and the other dimerization domain comprises a rapamycin analog binding sequence having at least 85% identity with FKBP.

64. The cell of claim 63, wherein one of said first and second dimerization domains comprises a rapamycin analog binding sequence from FKBP.

65. The cell of claim 63, wherein the other of said first and second dimerization domain comprises a rapamycin analog binding sequence from FRP.

66. The cell of claim 17, wherein said first and second chimeric protein both comprise an intracellular co-stimulatory domain derived from the same protein or different endogenous proteins.

67. The cell of claim 17, wherein said cell is selected from an NK cell or T cell.

68. A nucleic acid encoding the first and second chimeric proteins of claim 17.

69. A vector comprising the nucleic acid of claim 68.

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58. A chimeric membrane protein complex, comprising:

(i) a first chimeric membrane protein comprising an extracellular domain, said extracellular domain comprising a CD3 gamma, delta, or epsilon domain, a transmembrane domain and a first intracellular dimerization domain, wherein said CD3 gamma, delta, or epsilon domain comprises an extracellular domain derived from the extracellular domain of CD3 gamma, delta, or epsilon, and an intracellular co-stimulatory domain; and

(ii) a second chimeric membrane protein comprising an extracellular domain comprising an antigen binding domain, a transmembrane domain, an intracellular co-stimulatory domain and a second intracellular dimerization domain.

59. A chimeric membrane protein, comprising:

a chimeric membrane protein comprising an extracellular domain, said extracellular domain comprising;

(i) antigen binding domain,

(ii) a CD3 gamma, delta, or epsilon domain, wherein said CD3 gamma, delta, or epsilon domain comprises an extracellular domain derived from the extracellular domain of CD3 gamma, delta, or epsilon,

(iii) a transmembrane domain; and

(iv) an intracellular co-stimulatory domain.