Methods for improving the efficacy and expansion of chimeric antigen receptor-expressing cells

• US 11,896,614 B2
• Filed: 04/15/2016
• Issued: 02/13/2024
• Est. Priority Date: 04/17/2015
• Status: Active Grant

• Current Assignee: Novartis Ag, Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Original Assignee: Novartis Ag
• Inventors: Barrett, David M., Bedoya, Felipe, Ghassemi, Saba, June, Carl H., Levine, Bruce L., Melenhorst, Jan J., Milone, Michael C., Powell, Jr., Daniel J., Singh, Nathan Amar, Zheng, Zoe
• Primary Examiner(s): Moloye, Titilayo
• Application Number:
  US15/567,156
• Publication Number:
  US 20180133296A1
• Time in Patent Office: 2,860 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 2239/13 Antibody-based
• CPC Class Codes: A61K 2239/31 characterized by the route of administration
• CPC Class Codes: A61K 2239/38 characterised by the dose, timing or administration schedule
• CPC Class Codes: A61K 2239/59 Reproductive system, e.g. uterus, ovaries, cervix or testes
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 35/17 Lymphocytes; B-cells; T-cells; Natural killer cells; Interferon-activated or cytokine-activated lymphocytes when activated by a specific antigen A61K39/00
• CPC Class Codes: A61K 38/50 acting on carbon-nitrogen bonds, other than peptide bonds (3.5), e.g. asparaginase
• 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 39/4631 Chimeric Antigen Receptors [CAR]
• CPC Class Codes: A61K 39/464412 CD19 or B4
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 14/4748 Tumour specific antigens; Tumour rejection antigen precursors [TRAP], e.g. MAGE
• 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 14/70578 : NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95 NGF-receptor C07K14/71, TNF-receptor C07K14/7151
• 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 2317/622 : Single chain antibody (scFv)
• CPC Class Codes: C07K 2319/00 : Fusion polypeptide
• CPC Class Codes: C07K 2319/03 : containing a transmembrane segment
• 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: C12N 2501/2302 : Interleukin-2 (IL-2)
• CPC Class Codes: C12N 2501/2305 : Interleukin-5 (IL-5)
• CPC Class Codes: C12N 2501/2307 : Interleukin-7 (IL-7)
• CPC Class Codes: C12N 2501/2318 : Interleukin-18 (IL-18)
• CPC Class Codes: C12N 5/0636 : T lymphocytes
• CPC Class Codes: C12Y 305/01001 : Asparaginase (3.5.1.1)

Abstract

The invention provides methods of making immune effector cells (e.g., T cells, NK cells) that can be engineered to express a chimeric antigen receptor (CAR), compositions and reaction mixtures comprising the same, and methods of treatment using the same.

Claims

1. A method of making an immune effector cell population comprising an immune effector cell comprising a chimeric antigen receptor (“

CAR”

), comprising acquiring a human immune effector cell population from a pediatric subject having a leukemia, wherein the subject has not been treated with cyclophosphamide or cytarabine,

thereby making an immune effector cell population suitable for use in a CAR therapy, wherein the acquired immune effector cell population includes;

(a) one or more of at least 20% naï

ve T cells, at least 2% stem central memory T cells, and at least 4% central memory T cells;

(b) at least 50% central memory T cells;

(c) less than 55% effector memory and terminal effector T cells combined;

or

(d) two or more of (a), (b) and (c), wherein the combined percentages of the two or more of (a), (b), and (c) do not yield a percentage that exceeds 100%; and

wherein the method further comprises introducing into the immune effector cell population a nucleic acid encoding a CAR; and

wherein the immune effector cell population shows an increase in one or more of;

(i) ex-vivo expansion of the immune effector cell population, (ii) the efficacy of the immune effector cell population for therapy, or (iii) the yield of the immune effector cell population, relative to a population of immune effector cells acquired from a pediatric subject having a leukemia who has been treated with cyclophosphamide or cytarabine.

2. The method of claim 1, wherein the CAR therapy comprises a CD19 CAR molecule comprising a sequence set forth in any one of SEQ ID NOs:

39-102, 107-121, and 362.

3. The method of claim 1, wherein the subject is 18 years of age of younger or is 10 years of age or younger.

4. The method of claim 1, wherein the leukemia is chosen from one or more of:

a B-cell acute lymphoblastic leukemia (B-ALL), T-cell acute lymphoblastic leukemia (T-ALL), acute lymphoblastic leukemia (ALL), chronic myelogenous leukemia (CML), chronic lymphocytic leukemia (CLL), B cell promyelocytic leukemia, and hairy cell leukemia.

5. The method of claim 4, wherein:

(i) the leukemia is a CLL or an ALL;

(ii) the subject is classified as having standard-risk, high-risk, or very high-risk ALL;

(iii) the subject does not have a lymphoma;

or

(iv) the subject does not have a relapsed leukemia.

6. The method of claim 1, further comprising acquiring the immune effector cell population from the pediatric subject according to the timing of administration of chemotherapy to the subject in relation to the timing of the immune effector cell acquisition, wherein the chemotherapy or cycle of chemotherapy comprises one or more of an induction, a consolidation, an interim maintenance, a delayed intensification, or a maintenance therapy cycle, and wherein:

(i) the immune effector cell population is acquired from the subject before the subject has undergone a consolidation cycle of chemotherapy or an induction cycle of chemotherapy;

(ii) wherein the immune effector cell population is acquired from the subject before the subject has undergone a consolidation cycle of chemotherapy;

(iii) the subject is classified as having a high-risk or very high-risk cancer and the cells are harvested before the subject has undergone a consolidation cycle;

(iv) the immune effector cell population is acquired from the subject before the subject has undergone a delayed intensification cycle;

or

(v) the subject is classified as having a high-risk or very high-risk cancer and the cells are harvested before the subject has undergone a delayed intensification cycle.

7. The method of claim 6, wherein:

(a) one or more chemotherapy cycles or drugs are chosen from the group consisting of;

(i) vincristine, dexamethasone, PEG-L-asparaginase, daunorubicin;

(ii) vincristine, 6-mercaptopurine, cyclophosphamide, cytarabine, PEG-L-asparaginase;

(iii) vincristine, methotrexate, 6-mercaptopurine;

(iv) vincristine, dexamethasone, doxorubicin, PEG-L-asparaginase, cyclophosphamide, cytarabine, 6-thioguanine;

(v) vincristine, methotrexate, and

(vi) vincristine, dexamethasone, prednisone 6-mercaptopurine, methotrexate;

(b) the chemotherapy comprises a drug and dosing regimen for the treatment of subject classified as having standard-risk cancer is chosen from the group consisting of;

(i) vincristine 6 mg/m², dexamethasone 6 mg/m², PEG-L-asparaginase 2500 U/m²;

(ii) vincristine 1.5 mg/m², 6-mercaptopurine 75 mg/m²;

(iii) vincristine 7.5 mg/m², methotrexate 500 mg/m²;

(iv) vincristine 4.5 mg/m², dexamethasone 10 mg/m², doxorubicin 75 mg/m², PEG-L-asparaginase 2500 U/m², cyclophosphamide 1 g/m², cytarabine 60 mg/m², 6-thioguanine 60 mg/m²;

(v) vincristine 7.5 mg/m², methotrexate 500 mg/m²; and

(vi) vincristine 1.5 mg/m², dexamethasone 6 mg/m², 6-mercaptopurine 75 mg/m², methotrexate 20 mg/m²;

(c) the chemotherapy comprises a drug and dosing regimen for the treatment of subject classified as having high-risk cancer is chosen from the group consisting of;

(i) vincristine 6 mg/m², dexamethasone 6 mg/m², PEG-L-asparaginase 2500 U/m², daunorubicin 100 mg/m²;

(ii) vincristine 1.5 mg/m², 6-mercaptopurine 60 mg/m², cyclophosphamide 1 g/m², cytarabine 75 mg/m², PEG-L-asparaginase 2500 mg/m²;

(iii) vincristine 7.5 mg/m², methotrexate 20 mg/m², 6-mercaptopurine 25 mg/m²;

(iv) vincristine 7.5 mg/m², dexamethasone 10 mg/m², doxorubicin 75 mg/m², PEG-L-asparaginase 5000 U/m², cyclophosphamide 1 g/m², cytarabine 60 mg/m², 6-thioguanine 60 mg/m²; and

(v) vincristine 1.5 mg/m², prednisone 40 mg/m², 6-mercaptopurine 75 mg/m², methotrexate 20 mg/m²;

or

(d) the chemotherapy comprises one or more of vincristine, dexamethasone, PEG-L-asparaginase, daunorubicin, 6-mercaptopurine, cyclophosphamide, cytarabine, methotrexate, doxorubicin, 6-thioguanine, and prednisone.

8. The method of claim 1, wherein the immune effector cell population suitable for use in a CAR therapy comprises a higher number of less differentiated T cells.

9. The method of claim 1, wherein:

(i) the immune effector cell population suitable for use in a CAR therapy shows an increased absolute T cell count (ATC), compared to a reference value;

or

(ii) the immune effector cell population suitable for use in a CAR therapy comprises one or more of an absolute T cell count of at least 400 cells/microliter, an absolute naï

ve T cell count of at least 200 cells/microliter, an absolute stem central memory T cell count of at least 20 cells/microliter, or an absolute central memory T cell count of at least 40 cells/microliter.

10. The method of claim 1, wherein the immune effector cell population suitable for use in a CAR therapy is selected based upon the expression of one or more markers chosen from CCR7, CD62L, CD45RO, and CD95.

11. The method of claim 1, wherein the naï

ve T cells are identified based upon an expression pattern of CCR7+, CD62L+, CD45RO−

, CD95−

, wherein the stem central memory T cells are identified based upon an expression pattern of CCR7+, CD62L+, CD45RO−

, CD95+, and wherein the central memory T cells are identified based upon an expression pattern of CCR7+, CD62L+, CD45RO+, CD95+.

12. The method of claim 1, wherein the immune effector cell population has been selected based upon the expression of one or more markers chosen from CD3, CD28, CD4, CD8, CD45RA, and CD45RO.

13. The method of claim 1, further comprising:

activating the immune effector cell population, or transducing the immune effector cell population with a viral vector comprising a nucleic acid encoding a CAR.

14. The method of claim 1, wherein the immune effector cells are acquired from the subject prior to introduction of a CAR molecule into the immune effector cell.

15. The method of claim 1, wherein the CAR is a CD19 CAR.