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EP-4739323-A1 - <SMALLCAPS/>? ? ?EX VIVO? ? ? ? ?GENERATION OF IMMUNE EFFECTOR CELLS FROM APHERESIS MATERIAL INTERMEDIATES

EP4739323A1EP 4739323 A1EP4739323 A1EP 4739323A1EP-4739323-A1

Abstract

Methods and compositions are provided herein for generating tumor targeted lymphocytes and/or tumor infiltrating lymphocytes and/or tumor targeted natural killer (NK) cells for use in the treatment of a cancer or an infectious disease. Also provided herein are methods of making and using the same.

Inventors

  • SOON-SHIONG, PATRICK

Assignees

  • ImmunityBio, Inc.

Dates

Publication Date
20260513
Application Date
20240702

Claims (20)

  1. 1. A method of generating tumor targeted lymphocytes for use in the treatment of a cancer or infectious disease, the method comprising: i. performing therapeutic apheresis on a subject with cancer or infectious disease; ii. purifying a CD3+ T cell fraction from the apheresis product, wherein the remaining apheresis product comprises a CD3- fraction; iii. purifying CD14+ Monocytic cells from the CD3- fraction, wherein the remaining apheresis product comprises a CD3-/CD14- fraction; iv. differentiating the CD14+ Monocytes into dendritic cells (DC), and exposing the DC to an antigenic peptide or an adenovirus encoding an antigenic peptide sequence, wherein the DC MHC-I or MHC-II present the peptide sequence or a portion thereof, thereby activating the DC; v. exposing the purified CD3+ T cells to the activated DC, thereby expanding the T cells; vi. purifying the expanded T cells.
  2. 2. The method of claim 1, wherein the T cells are expanded in the presence of IL- 15 or an agonist derivative thereof.
  3. 3. The method of any one of claims 1-2, wherein the T cells are genetically modified to express endoplasmic reticulum localized IL-15 (erIL-15).
  4. 4. The method of any one of claims 1-3, wherein the T cells are genetically modified to express a chimeric antigen receptor (CAR), and wherein the CAR targets a tumor antigen or a checkpoint inhibitor.
  5. 5. The method of any one of claims 1-4, wherein the adenovirus comprises an Ad5 adenovirus.
  6. 6. The method of any one of claims 1-5, wherein the dendritic cells are further exposed to a modified RNA.
  7. 7. The method of any one of claims 1-6, wherein the dendritic cells are further exposed to a lenti virus.
  8. 8. The method of any one of claims 1-2, wherein the dendritic cells are further exposed to a peptide pool of neoeptiopes.
  9. 9. A method of generating tumor targeted natural killer (NK) cell for use in the treatment of a cancer or infectious disease, the method comprising: i. performing therapeutic apheresis on a subject with cancer or infectious disease; ii. purifying a CD3+ T cell fraction from the apheresis product, wherein the remaining apheresis product comprises a CD3- fraction; iii. purifying CD 14+ Monocytic cells from the CD3- fraction of the apheresis, wherein the remaining apheresis product comprises a CD3-/CD14- fraction; iv. expanding NK cells from the CD3- CD14- fraction of the apheresis; v. differentiating the CD14+ Monocytes into dendritic cells (DC), and exposing the DC to an antigenic peptide or an adenovirus encoding an antigenic peptide sequence, wherein the DC MHC-I or MHC-II present the peptide sequence or a portion thereof, thereby activating the DC; vi. exposing the purified CD3+ T cells to the activated DC, thereby expanding the T cells; vii. purifying the expanded T cells. viii. isolating from the expanded T cells at least one nucleic acid encoding an a and chain of a T cell receptor (TCR), and fusing the nucleic acid to the 5’ end of a second nucleic acid encoding a transmembrane domain and an intracellular signaling domain of a chimeric antigen receptor (CAR), wherein the fused nucleic acid encodes a TCR CAR; ix. transfecting the enriched and expanded NK cells with a nucleic acid encoding the TCR CAR.
  10. 10. The method of claim 9, wherein the NK cells comprise memory cytokine enriched NK cells (M-CENK).
  11. 11. The method of any one of claims 9-10, wherein the M-CENK are genetically modified to express CD 16.
  12. 12. The method of any one of claims 10-11, wherein the M-CENK cells are further genetically modified to express a CAR, wherein the CAR targets a tumor antigen or a checkpoint inhibitor.
  13. 13. The method of any one of claims 9-12, wherein the NK cells comprise NK-92 cells, and optionally wherein the NK-92 cells comprise CD 16.
  14. 14. The method of claim 13, wherein the NK-92 cells are further genetically modified to express a second CAR.
  15. 15. A method of expanding tumor infiltrating lymphocytes for use in the treatment of a cancer or infectious disease, the method comprising: i. performing therapeutic apheresis on a subject with cancer or infectious disease; ii. purifying a CD3+ T cell fraction from the apheresis product, wherein the remaining apheresis product comprises a CD3- fraction; iii. purifying CD14+ Monocytic cells from the CD3- fraction of the apheresis, wherein the remaining apheresis product comprises a CD3-/CD14- fraction; iv. expanding NK cells from the CD3-/CD14- fraction of the apheresis; v. differentiating the CD14+ Monocytes into dendritic cells (DC), and exposing the DC to an antigenic peptide or an adenovirus encoding an antigenic peptide sequence, wherein the DC MHC-I or MHC-II present the peptide sequence or a portion thereof, thereby activating the DC; vi. exposing the purified CD3+ T cells to the activated DC, thereby expanding the T cells; vii. purifying the expanded T cells; viii. purifying CD3+ TIL from a solid tumor and exposing the TIL to the activated DC, thereby expanding the TIL; ix. purifying the expanded TIL.
  16. 16. A pharmaceutical composition comprising tumor targeted lymphocytes for use in the treatment of a cancer or infectious disease, the composition comprising 1) activated dendritic cells (DC), wherein the DC are differentiated from patient apheresis-derived CD14+ monocytes; 2) an adenovirus encoding an antigenic peptide sequence, wherein the DC are activated upon exposure to the adenovirus; and 3) patient apheresis-derived CD3+ T cells, wherein the CD3+ T cells are exposed to the activated DC, thereby activating and expanding the T cells.
  17. 17. The pharmaceutical composition of claim 16, wherein DC MHC-I or MHC-II present the antigenic peptide sequence or a portion thereof, thereby activating the DC.
  18. 18. The pharmaceutical composition of any one of claims 16-17, wherein the activated CD3+ T cells comprise a T cell receptor (TCR) with specificity for the MHC-presented antigenic peptide sequence on the DC.
  19. 19. The pharmaceutical composition of any one of claims 16-18, further comprising Natural Killer (NK) cells.
  20. 20. The pharmaceutical composition of claim 19, wherein the NK cells are expanded from the patient apheresis.

Description

EX VIVO GENERATION OE IMMUNE EFFECTOR CELLS FROM APHERESIS MATERIAL INTERMEDIATES This application claims the benefit of priority to U.S. Provisional Application Nos.: 63/512,032 filed on July 5, 2023; 63/515,528 filed on July 25, 2023, and 63/605,326 filed on December 1, 2023. Each of the above applications are incorporated by reference in their entirety. Field of the Invention [0001] The field of the invention is immunotherapy technologies. Background [0002] The background description includes information that may be useful in understanding the present disclosure. It is not an admission that any of the information provided herein is prior art or relevant to the presently claimed invention, or that any publication specifically or implicitly referenced is prior art. [0003] All publications and patent applications herein are incorporated by reference to the same extent as if each individual publication or patent application were specifically and individually indicated to be incorporated by reference. Where a definition or use of a term in an incorporated reference is inconsistent or contrary to the definition of that term provided herein, the definition of that term provided herein applies and the definition of that term in the reference does not apply. [0004] Autologous or allogenic T-cells and natural killer cells provide meaningful benefit for otherwise refractory malignancies. Apheresis generates the starting material for T cell or NK cell manufacturing. Because NK cells comprise only a small fraction of lymphocytes (~l-20%), methods have been developed either to enrich them from large volumes of peripheral blood, such as an apheresis product, or to expand the NK cell population from a smaller number of blood or stem cells. As clinical indications for T cells and NK cells are expanding, researchers are trying to develop new and improved ways of obtaining these cells from peripheral blood mononuclear cells (PBMCs). [0005] Thus, there remains a need in the art for improved methods of isolating and expanding T cells and NK cells for treatment of a cancer or infectious disease in a patient in need thereof. Summary [0006] The inventive subject matter provides compositions and methods for generating tumor targeted lymphocytes and/or tumor infiltrating lymphocytes and/or tumor targeted natural killer (NK) cells for use in the treatment of a cancer or an infectious disease. [0007] In an embodiment, the inventive subject matter comprises tumor targeted lymphocytes for use in the treatment of a cancer or infectious disease. The tumor targeted lymphocytes are generated by a method comprising the steps of performing therapeutic apheresis on a subject with cancer or infectious disease; purifying a CD3+ T cell fraction from the apheresis product, wherein the remaining apheresis product comprises a CD3- fraction; purifying CD14+ Monocytic cells from the CD3- fraction, wherein the remaining apheresis product comprises a CD3-/CD14- fraction; differentiating the CD14+ Monocytes into dendritic cells (DC), and exposing the DC to an antigenic peptide or an adenovirus encoding an antigenic peptide sequence, wherein the DC MHC-I or MHC-II present the peptide sequence or a portion thereof, thereby activating the DC; exposing the purified CD3+ T cells to the activated DC, thereby expanding the T cells; and purifying the expanded T cells. [0008] In an embodiment, the inventive subject matter comprises tumor targeted natural killer (NK) cells for use in the treatment of a cancer or infectious disease. The NK cells are generated by a method comprising the steps of performing therapeutic apheresis on a subject with cancer or infectious disease; purifying a CD3+ T cell fraction from the apheresis product, wherein the remaining apheresis product comprises a CD3- fraction; purifying CD14+ Monocytic cells from the CD3- fraction of the apheresis, wherein the remaining apheresis product comprises a CD3- /CD14- fraction; expanding NK cells from the CD3- CD14- fraction of the apheresis; differentiating the CD14+ Monocytes into dendritic cells (DC), and exposing the DC to an antigenic peptide or an adenovirus encoding an antigenic peptide sequence, wherein the DC MHC-I or MHC-II present the peptide sequence or a portion thereof, thereby activating the DC; exposing the purified CD3+ T cells to the activated DC, thereby expanding the T cells; purifying the expanded T cells; isolating from the expanded T cells at least one nucleic acid encoding an a and P chain of a T cell receptor (TCR), and fusing the nucleic acid to the 5’ end of a second nucleic acid encoding a transmembrane domain and an intracellular signaling domain of a chimeric antigen receptor (CAR), wherein the fused nucleic acid encodes a TCR CAR; and transfecting the enriched and expanded NK cells with a nucleic acid encoding the TCR CAR. [0009] In an embodiment, the inventive subject matter comprises a method of expanding tumor infdtrating lymphocytes for use in the treatment