Bcma chimeric antigen receptors

Abstract

The invention provides improved compositions for adoptive T cell therapies for B cell related conditions.

1 . A chimeric antigen receptor (CAR) comprising: an extracellular domain that comprises a humanized anti-BCMA (B cell maturation antigen) antibody or antigen binding fragment thereof that binds one or more epitopes of a human BCMA polypeptide; a transmembrane domain, one or more intracellular co-stimulatory signaling domains, and a primary signaling domain. 
     
     
         2 . The CAR of  claim 1 , wherein the humanized anti-BCMA antibody or antigen binding fragment that binds the human BCMA polypeptide is selected from the group consisting of: a Camel Ig, Ig NAR, Fab fragments, Fab′ fragments, F(ab)′2 fragments, F(ab)′3 fragments, Fv, single chain Fv antibody (“scFv”), bis-scFv, (scFv)2, minibody, diabody, triabody, tetrabody, disulfide stabilized Fv protein (“dsFv”), and single-domain antibody (sdAb, Nanobody). 
     
     
         3 . The CAR of  claim 2 , wherein the humanized anti-BCMA antibody or antigen binding fragment that binds the human BCMA polypeptide is an scFv. 
     
     
         4 . The CAR of any one of  claims 1  to  3 , wherein the humanized anti-BCMA antibody or antigen binding fragment thereof comprises one or more CDRs as set forth in any one of SEQ ID NOs: 1-3. 
     
     
         5 . The CAR of any one of  claims 1  to  4 , wherein the humanized anti-BCMA antibody or antigen binding fragment thereof comprises one or more CDRs as set forth in any one of SEQ ID NOs: 4-6. 
     
     
         6 . The CAR of any one of  claims 1  to  5 , wherein the humanized anti-BCMA antibody or antigen binding fragment thereof comprises a variable light chain sequence as set forth in any one of SEQ ID NOs: 7-9. 
     
     
         7 . The CAR of  claim 6 , wherein the variable light chain sequence comprises the CDR sequences set forth in SEQ ID NOs: 1-3. 
     
     
         8 . The CAR of any one of  claims 1  to  7 , wherein the humanized anti-BCMA antibody or antigen binding fragment thereof comprises a variable heavy chain sequence as set forth in any one of SEQ ID NOs: 10-14. 
     
     
         9 . The CAR of  claim 8 , wherein the variable heavy chain sequence comprises the CDR sequences set forth in SEQ ID NOs: 4-6 
     
     
         10 . The CAR of any one of  claims 1  to  9 , wherein the transmembrane domain is from a polypeptide selected from the group consisting of: alpha, beta or zeta chain of the T-cell receptor, CD3ε, CD3ζ, CD4, CD5, CD8α, CD9, CD 16, CD22, CD27, CD28, CD33, CD37, CD45, CD64, CD80, CD86, CD 134, CD137, CD152, CD 154, and PD1. 
     
     
         11 . The CAR of any one of  claims 1  to  9 , wherein the transmembrane domain is from a polypeptide selected from the group consisting of: CD8α; CD4, CD45, PD1, and CD152. 
     
     
         12 . The CAR of any one of  claims 1  to  11 , wherein the transmembrane domain is from CD8α. 
     
     
         13 . The CAR of any one of  claims 1  to  11 , wherein the transmembrane domain is from PD1. 
     
     
         14 . The CAR of any one of  claims 1  to  11 , wherein the transmembrane domain is from CD152. 
     
     
         15 . The CAR of any one of  claims 1  to  14 , wherein the one or more co-stimulatory signaling domains are from a co-stimulatory molecule selected from the group consisting of: CARD11, CD2, CD7, CD27, CD28, CD30, CD40, CD54 (ICAM), CD83, CD134 (OX40), CD137 (4-1BB), CD150 (SLAMF1), CD152 (CTLA4), CD223 (LAG3), CD270 (HVEM), CD273 (PD-L2), CD274 (PD-L1), CD278 (ICOS), DAP10, LAT, NKD2C SLP76, TRIM, and ZAP70. 
     
     
         16 . The CAR of any one of  claims 1  to  14 , wherein the one or more co-stimulatory signaling domains are from a co-stimulatory molecule selected from the group consisting of: CD28, CD134, and CD137. 
     
     
         17 . The CAR of any one of  claims 1  to  14 , wherein the one or more co-stimulatory signaling domains are from a co-stimulatory molecule selected from the group consisting of: CD28, CD134, and CD137. 
     
     
         18 . The CAR of any one of  claims 1  to  14 , wherein the one or more co-stimulatory signaling domains is from CD28. 
     
     
         19 . The CAR of any one of  claims 1  to  14 , wherein the one or more co-stimulatory signaling domains is from CD134. 
     
     
         20 . The CAR of any one of  claims 1  to  14 , wherein the one or more co-stimulatory signaling domains is from CD137. 
     
     
         21 . The CAR of any one of  claims 1  to  20 , further comprising a hinge region polypeptide. 
     
     
         22 . The CAR of  claim 21 , wherein the hinge region polypeptide comprises a hinge region of CD8α. 
     
     
         23 . The CAR of  claim 21 , wherein the hinge region polypeptide comprises a hinge region of PD1. 
     
     
         24 . The CAR of  claim 21 , wherein the hinge region polypeptide comprises a hinge region of CD152. 
     
     
         25 . The CAR of any one of  claims 1  to  24 , further comprising a spacer region. 
     
     
         26 . The CAR of  claim 25 , wherein the spacer region polypeptide comprises CH2 and CH3 regions of IgG1 or IgG4. 
     
     
         27 . The CAR of any one of  claims 1  to  26 , further comprising a signal peptide. 
     
     
         28 . The CAR of  claim 27 , wherein the signal peptide comprises an IgG1 heavy chain signal polypeptide, a CD8α signal polypeptide, or a human GM-CSF receptor alpha signal polypeptide. 
     
     
         29 . The CAR of any one of  claims 1  to  28 , comprising an amino acid sequence as set forth in any one of SEQ ID NOs: 15-29, 71, and 73. 
     
     
         30 . The CAR of any one of  claims 1  to  29 , comprising an amino acid sequence as set forth in SEQ ID NO: 15. 
     
     
         31 . The CAR of any one of  claims 1  to  29 , comprising an amino acid sequence as set forth in SEQ ID NO: 16. 
     
     
         32 . The CAR of any one of  claims 1  to  29 , comprising an amino acid sequence as set forth in SEQ ID NO: 17. 
     
     
         33 . The CAR of any one of  claims 1  to  29 , comprising an amino acid sequence as set forth in SEQ ID NO: 18. 
     
     
         34 . The CAR of any one of  claims 1  to  29 , comprising an amino acid sequence as set forth in SEQ ID NO: 19. 
     
     
         35 . The CAR of any one of  claims 1  to  29 , comprising an amino acid sequence as set forth in SEQ ID NO: 20. 
     
     
         36 . The CAR of any one of  claims 1  to  29 , comprising an amino acid sequence as set forth in SEQ ID NO: 21. 
     
     
         37 . The CAR of any one of  claims 1  to  29 , comprising an amino acid sequence as set forth in SEQ ID NO: 22. 
     
     
         38 . The CAR of any one of  claims 1  to  29 , comprising an amino acid sequence as set forth in SEQ ID NO: 23. 
     
     
         39 . The CAR of any one of  claims 1  to  29 , comprising an amino acid sequence as set forth in SEQ ID NO: 24. 
     
     
         40 . The CAR of any one of  claims 1  to  29 , comprising an amino acid sequence as set forth in SEQ ID NO: 25. 
     
     
         41 . The CAR of any one of  claims 1  to  29 , comprising an amino acid sequence as set forth in SEQ ID NO: 26. 
     
     
         42 . The CAR of any one of  claims 1  to  29 , comprising an amino acid sequence as set forth in SEQ ID NO: 27. 
     
     
         43 . The CAR of any one of  claims 1  to  29 , comprising an amino acid sequence as set forth in SEQ ID NO: 28. 
     
     
         44 . The CAR of any one of  claims 1  to  29 , comprising an amino acid sequence as set forth in SEQ ID NO: 29. 
     
     
         45 . The CAR of any one of  claims 1  to  29 , comprising an amino acid sequence as set forth in SEQ ID NO: 71. 
     
     
         46 . The CAR of any one of  claims 1  to  29 , comprising an amino acid sequence as set forth in SEQ ID NO: 73. 
     
     
         47 . A polynucleotide encoding a CAR of any one of  claims 1  to  46 . 
     
     
         48 . A polynucleotide encoding a CAR, wherein the polynucleotide sequence is set forth in any one of SEQ ID NOs: 30-44, 70, and 72. 
     
     
         49 . A vector comprising the polynucleotide of  claim 47  or  claim 48 . 
     
     
         50 . The vector of  claim 49 , wherein the vector is an expression vector. 
     
     
         51 . The vector of  claim 49 , wherein the vector is an episomal vector. 
     
     
         52 . The vector of  claim 49 , wherein the vector is a viral vector. 
     
     
         53 . The vector of  claim 49 , wherein the vector is a retroviral vector. 
     
     
         54 . The vector of  claim 49 , wherein the vector is a lentiviral vector. 
     
     
         55 . The vector of  claim 49 , wherein the lentiviral vector is selected from the group consisting essentially of: human immunodeficiency virus 1 (HIV-1); human immunodeficiency virus 2 (HIV-2), visna-maedi virus (VMV) virus; caprine arthritis-encephalitis virus (CAEV); equine infectious anemia virus (EIAV); feline immunodeficiency virus (FIV); bovine immune deficiency virus (BIV); and simian immunodeficiency virus (SIV). 
     
     
         56 . The vector according to any one of  claims 52  to  55 , comprising a left (5′) retroviral LTR, a Psi (Ψ) packaging signal, a central polypurine tract/DNA flap (cPPT/FLAP), a retroviral export element; a promoter operably linked to the polynucleotide of  claim 26  or  claim 27 ; and a right (3′) retroviral LTR. 
     
     
         57 . The vector of  claim 56 , further comprising a heterologous polyadenylation sequence. 
     
     
         58 . The vector of  claim 56  or  claim 57 , further comprising a hepatitis B virus posttranscriptional regulatory element (HPRE) or woodchuck post-transcriptional regulatory element (WPRE). 
     
     
         59 . The vector of any one of  claims 56  to  58 , wherein the promoter of the 5′ LTR is replaced with a heterologous promoter. 
     
     
         60 . The vector of  claim 59 , wherein the heterologous promoter is a cytomegalovirus (CMV) promoter, a Rous Sarcoma Virus (RSV) promoter, or an Simian Virus 40 (SV40) promoter. 
     
     
         61 . The vector of any one of  claims 56  to  60 , wherein the 5′ LTR or 3′ LTR is a lentivirus LTR. 
     
     
         62 . The vector of any one of  claims 56  to  61 , wherein the 3′ LTR comprises one or more modifications. 
     
     
         63 . The vector of any one of  claims 56  to  62 , wherein the 3′ LTR comprises one or more deletions. 
     
     
         64 . The vector of any one of  claims 56  to  63 , wherein the 3′ LTR is a self-inactivating (SIN) LTR. 
     
     
         65 . The vector of any one of  claims 56  to  64 , wherein the polyadenylation sequence is a bovine growth hormone polyadenylation or signal rabbit β-globin polyadenylation sequence. 
     
     
         66 . The vector of any one of  claims 59  to  65 , wherein the polynucleotide of  claim 57  or  claim 58  comprises an optimized Kozak sequence. 
     
     
         67 . The vector of any one of  claims 56  to  66 , wherein the promoter operably linked to the polynucleotide of  claim 57  or  claim 58  is selected from the group consisting of: a cytomegalovirus immediate early gene promoter (CMV), an elongation factor 1 alpha promoter (EF1-α), a phosphoglycerate kinase-1 promoter (PGK), a ubiquitin-C promoter (UBQ-C), a cytomegalovirus enhancer/chicken beta-actin promoter (CAG), polyoma enhancer/herpes simplex thymidine kinase promoter (MC1), a beta actin promoter (β-ACT), a simian virus 40 promoter (SV40), and a myeloproliferative sarcoma virus enhancer, negative control region deleted, d1587rev primer-binding site substituted (MND) promoter. 
     
     
         68 . An immune effector cell comprising the vector of any one of  claims 49  to  67 . 
     
     
         69 . The immune effector cell of  claim 68 , wherein the immune effector cell is selected from the group consisting of: a T lymphocyte and a natural killer (NK) cell. 
     
     
         70 . A composition comprising the immune effector cell of  claim 68  or  claim 69  and a physiologically acceptable excipient. 
     
     
         71 . A method of generating an immune effector cell comprising a CAR according to any one of  claims 1  to  46  comprising introducing into an immune effector cell the vector of an one of  claims 49  to  67 . 
     
     
         72 . The method of  claim 71 , further comprising stimulating the immune effector cell and inducing the cell to proliferate by contacting the cell with antibodies that bind CD3 and antibodies that bind to CD28; thereby generating a population of immune effector cells. 
     
     
         73 . The method of  claim 72 , wherein the immune effector cell is stimulated and induced to proliferate before introducing the vector. 
     
     
         74 . The method of  claim 72 , wherein the immune effector cells comprise T lymphocytes. 
     
     
         75 . The method of  claim 72 , wherein the immune effector cells comprise NK cells. 
     
     
         76 . A method of treating a B cell related condition in a subject in need thereof, comprising administering to the subject a therapeutically effect amount of the composition of  claim 70 . 
     
     
         77 . The method of  claim 76 , wherein the B cell related condition is multiple myeloma, non-Hodgkin's lymphoma, B cell proliferations of uncertain malignant potential, lymphomatoid granulomatosis, post-transplant lymphoproliferative disorder, an immunoregulatory disorder, rheumatoid arthritis, myasthenia gravis, idiopathic thrombocytopenia purpura, anti-phospholipid syndrome, Chagas' disease, Grave's disease, Wegener's granulomatosis, poly-arteritis nodosa, Sjogren's syndrome, pemphigus vulgaris, scleroderma, multiple sclerosis, anti-phospholipid syndrome, ANCA associated vasculitis, Goodpasture's disease, Kawasaki disease, autoimmune hemolytic anemia, and rapidly progressive glomerulonephritis, heavy-chain disease, primary or immunocyte-associated amyloidosis, or monoclonal gammopathy of undetermined significance. 
     
     
         78 . The method of  claim 76 , wherein the B cell related condition is a B cell malignancy. 
     
     
         79 . The method of  claim 78 , wherein the B cell malignancy is multiple myeloma (MM) or non-Hodgkin's lymphoma (NHL). 
     
     
         80 . The method of  claim 79 , wherein the MM is selected from the group consisting of: overt multiple myeloma, smoldering multiple myeloma, plasma cell leukemia, non-secretory myeloma, IgD myeloma, osteosclerotic myeloma, solitary plasmacytoma of bone, and extramedullary plasmacytoma. 
     
     
         81 . The method of  claim 79 , wherein the NHL is selected from the group consisting of: Burkitt lymphoma, chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL), diffuse large B-cell lymphoma, follicular lymphoma, immunoblastic large cell lymphoma, precursor B-lymphoblastic lymphoma, and mantle cell lymphoma. 
     
     
         82 . The method of  claim 76 , wherein the B cell related condition is a plasma cell malignancy. 
     
     
         83 . The method of  claim 76 , wherein the B cell related condition is an autoimmune disease. 
     
     
         84 . The method of  claim 83 , wherein the autoimmune disease is systemic lupus erythematosus. 
     
     
         85 . The method of  claim 83 , wherein the B cell related condition is rheumatoid arthritis. 
     
     
         86 . The method of  claim 83 , wherein the B cell related condition is idiopathic thrombocytopenia purpura, or myasthenia gravis, or autoimmune hemolytic anemia.