Methods and compositions for dosing of allogeneic chimeric antigen receptor t cells

Abstract

The present disclosure concerns dosages for the treatment of human patients susceptible to or diagnosed with a disease, such as cancer. Provided are methods for administering chimeric antigen receptor (CAR)-T cells. Also provided are compositions and articles of manufacture for use in the methods.

1 . A method of treating a subject who has refractory and/or relapsed Non-Hodgkin's Lymphoma, the method comprising administering to the subject at least one dose of allogeneic chimeric antigen receptor (CAR)-T cells (CAR-T cells) comprising an anti-human CD19 4-1BB/CD3zeta CAR, wherein the at least one dose is about 20×10 6  cells/dose to about 360×10 6  cells/dose. 
     
     
         2 . The method of  claim 1 , wherein the Non-Hodgkin's Lymphoma is large B-cell lymphoma. 
     
     
         3 . The method of  claim 1 , wherein the Non-Hodgkin's Lymphoma is follicular lymphoma. 
     
     
         4 . The method of  claim 1 , wherein the at least one dose is selected from the group consisting of about 20×10 6  cells/dose, about 40×10 6  cells/dose, about 80×10 6  cells/dose, about 120×10 6  cells/dose, about 240×10 6  cells/dose, and about 360×10 6  cells/dose. 
     
     
         5 .- 7 . (canceled) 
     
     
         8 . The method of  claim 1 , wherein the CAR-T cells comprise a mixture of CD52-deficient and CD-52 positive cells. 
     
     
         9 . The method of  claim 1 , wherein the CAR-T cells comprise the CAR of SEQ ID NO: 1. 
     
     
         10 . The method of  claim 1 , wherein the CAR-T cells comprise UCART19(CD19)CAR/RQR8+_TCRαβ−_T-cells. 
     
     
         11 . The method of  claim 1 , wherein the CAR-T cells do not express a safety switch. 
     
     
         12 .- 17 . (canceled) 
     
     
         18 . The method of  claim 1 , wherein the subject receives a first lymphodepletion regimen prior to administration of the at least one dose. 
     
     
         19 . The method of  claim 18 , wherein the first lymphodepletion regimen comprises administering fludarabine and cyclophosphamide. 
     
     
         20 . The method of  claim 18 , wherein the first lymphodepletion regimen comprises administering fludarabine, cyclophosphamide, and an anti-CD52 antibody. 
     
     
         21 .- 29 . (canceled) 
     
     
         30 . The method of  claim 20 , wherein fludarabine is administered at a dosage of about 30 mg/m 2 /day; cyclophosphamide is administered at a dosage of about 300 mg/m 2 /day; and CD52 antibody is administered at a dosage of about 10 to about 13 mg/day. 
     
     
         31 .- 32 . (canceled) 
     
     
         33 . The method of  claim 1 , wherein the subject receives a subsequent dose of the CAR-T cells. 
     
     
         34 .- 39 . (canceled) 
     
     
         40 . A pharmaceutical kit for treating a patient suffering from cancer, the kit comprising:
 (a) anti-CD19 CAR-T cells; and   (b) a CD52 antibody.   
     
     
         41 .- 46 . (canceled) 
     
     
         47 . A method of treatment, comprising:
 (a) administering to a subject having a disease a first dose of chimeric antigen receptor (CAR)-T cells (CAR-T cells), and   (b) administering to the subject a subsequent dose of CAR-T cells at a time point that is at least or more than about 28 days after and less than about 200 days after initiation of said administration in (a).   
     
     
         48 . (canceled) 
     
     
         49 . The method of  claim 47 , wherein the first dose comprises about 1×10 4  to about 5×10 8  total cells. 
     
     
         50 . (canceled) 
     
     
         51 . The method of  claim 47 , wherein the first dose comprises between 1×10 4  and 2×10 7  cells per kilogram body weight of the subject. 
     
     
         52 .- 54 . (canceled) 
     
     
         55 . The method of  claim 47 , wherein prior to step (a) or after step (a) and prior to step (b), the subject is administered a lymphodepletion regimen. 
     
     
         56 . The method of  claim 47 , wherein the lymphodepletion regimen comprises administering fludarabine, cyclophosphamide, and CD52 antibody to the subject between about 0 to 14 days prior to step (a) or step (b). 
     
     
         57 . The method of  claim 56 , wherein fludarabine is administered at a dosage of about 90 to 150 mg/m 2 ; cyclophosphamide is administered at a dosage of about 1000 to 4000 mg/m 2 ; and CD52 antibody is administered at a dosage of about 0.3 to 1 mg/kg. 
     
     
         58 .- 64 . (canceled) 
     
     
         65 . The method of  claim 47 , wherein the subsequent dose comprises about 1×10 5  to about 5×10 8  total cells. 
     
     
         66 .- 83 . (canceled) 
     
     
         84 . The method of  claim 47 , wherein the disease is a tumor or a cancer. 
     
     
         85 . The method of  claim 84 , wherein the cancer is leukemia or lymphoma. 
     
     
         86 . The method of  claim 84 , wherein the tumor or cancer is acute lymphoblastic leukemia (ALL), is chronic lymphocytic leukemia (CLL), HNSCC, non-Hodgkin's lymphoma, acute myeloid leukemia, diffuse large B-cell lymphoma (DLBCL), multiple myeloma, refractory follicular lymphoma, mantle cell lymphoma, indolent B cell lymphoma, B cell malignancies, cancers of the colon, lung, liver, breast, prostate, ovarian, skin, melanoma, bone, and brain cancer, ovarian cancer, epithelial cancers, renal cell carcinoma, pancreatic adenocarcinoma, Hodgkin lymphoma, cervical carcinoma, colorectal cancer, glioblastoma, neuroblastoma, Ewing sarcoma, medulloblastoma, osteosarcoma, synovial sarcoma, or mesothelioma. 
     
     
         87 . The method of  claim 86 , wherein the ALL is relapsed or refractory ALL. 
     
     
         88 . A method of treatment, comprising administering a subsequent dose of allogeneic chimeric antigen receptor (CAR)-T cells to a subject previously administered a first dose of allogeneic CAR-T cells, wherein: the subsequent dose of cells is administered at a time point that is at least or more than about 5 weeks after and less than about 24 weeks after initiation of the first dose. 
     
     
         89 .- 98 . (canceled) 
     
     
         99 . An article of manufacture, comprising:
 a plurality of sealable containers, each individually comprising a unit dose of allogeneic chimeric antigen receptor (CAR)-T cells for administration to a subject, said unit dose comprising about 1×10 6  to about 5×10 8  cells;   packaging material; and   a label or package insert comprising instructions for administering a plurality of said unit doses to the subject by carrying out a first administration and a subsequent administration, said first administration comprising delivering one of said unit doses to the subject and said subsequent administration comprising administering one or a plurality of said unit doses to the subject.   
     
     
         100 .- 101 . (canceled) 
     
     
         102 . A method of treating an adult subject who has refractory and/or relapsed CD19+ B-cell acute lymphoblastic leukemia, comprising administering to the subject at least one dose of allogeneic chimeric antigen receptor (CAR)-T cells (CAR-T cells) comprising an anti-human CD19 4-1BB/CD3zeta CAR, wherein the at least one dose is selected from the group consisting of about 6×10 5  cells/dose, 6×10 6  cells/dose, about 6-8×10 7  cells/dose, and about 1.8-2.4×10 8  cells/dose. 
     
     
         103 . A method of treating an pediatric subject who has refractory and/or relapsed CD19+ B-cell acute lymphoblastic leukemia, comprising administering to the subject at least one dose of allogeneic chimeric antigen receptor (CAR)-T cells (CAR-T cells) comprising an anti-human CD19 4-1BB/CD3zeta CAR, wherein the at least one dose is about 2-8×10 7  cells/dose. 
     
     
         104 . (canceled) 
     
     
         105 . The method of  claim 102 , wherein the CAR-T cells comprise a mixture of CD52-deficient and CD-52 positive cells. 
     
     
         106 . The method of  claim 102 , wherein the CAR-T cells express the CAR of SEQ ID NO: 1. 
     
     
         107 . The method of  claim 102 , wherein the CAR-T cells comprise UCART19(CD19CAR/RQR8+_TCRαβ−_T-cells). 
     
     
         108 .- 113 . (canceled) 
     
     
         114 . The method of  claim 102 , wherein the subject receives a first lymphodepletion regimen prior to administration of the at least one dose. 
     
     
         115 . The method of  claim 114 , wherein the first lymphodepletion regimen comprises administering fludarabine and cyclophosphamide. 
     
     
         116 . The method of  claim 114 , wherein the first lymphodepletion regimen comprises administering fludarabine, cyclophosphamide, and an anti-CD52 antibody. 
     
     
         117 . The method of  claim 116 , wherein fludarabine is administered at a dosage of about 30 to 150 mg/m 2 ; cyclophosphamide is administered at a dosage of about 300 to 4000 mg/m 2 ; and CD52 antibody is administered at a dosage of about 0.3 to 1 mg/kg. 
     
     
         118 . (canceled) 
     
     
         119 . The method of  claim 102 , wherein the subject receives a subsequent dose of the CAR-T cells. 
     
     
         120 .- 124 . (canceled) 
     
     
         125 . A method of producing a population of allogeneic chimeric antigen receptor (CAR)-T cells (CAR-T cells) directed to a target of interest comprising:
 (a) isolating peripheral blood mononuclear cells (PBMCs) from a healthy donor;   (b) activating the T-cells in the PBMCs;   (c) transducing the activated T-cells with a lentiviral vector, wherein the lentiviral vector is a self-inactivating recombinant vector expressing a CAR of interest;   (d) disrupting TCRαβ and CD-52 gene expression in a subset of the T-cells;   (e) expanding the population of T-cells; and   (f) enriching the population of T-cells for TCRαβ-negative cells;   thereby generating a population of allogeneic chimeric antigen receptor (CAR)-T cells (CAR-T cells).   
     
     
         126 .- 134 . (canceled) 
     
     
         135 . The method of  claim 103 , wherein the CAR-T cells comprise a mixture of CD52-deficient and CD-52 positive cells. 
     
     
         136 . The method of  claim 103 , wherein the CAR-T cells express the CAR of SEQ ID NO: 1. 
     
     
         137 . The method of  claim 103 , wherein the CAR-T cells comprise UCART19(CD19CAR/RQR8+_TCRαβ−_T-cells). 
     
     
         138 . The method of  claim 103 , wherein the subject receives a first lymphodepletion regimen prior to administration of the at least one dose. 
     
     
         139 . The method of  claim 138 , wherein the first lymphodepletion regimen comprises administering fludarabine and cyclophosphamide. 
     
     
         140 . The method of  claim 138 , wherein the first lymphodepletion regimen comprises administering fludarabine, cyclophosphamide, and an anti-CD52 antibody. 
     
     
         141 . The method of  claim 140 , wherein fludarabine is administered at a dosage of about 150 mg/m 2 ; cyclophosphamide is administered at a dosage of about 120 mg/kg; and CD52 antibody is administered at a dosage of about 1 mg/kg or a flat dose of about 40 mg. 
     
     
         142 . The method of  claim 103 , wherein the subject receives a subsequent dose of the CAR-T cells.