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EP-4735026-A1 - COMBINATION THERAPY USING CAR T CELLS AND KLOTHO

EP4735026A1EP 4735026 A1EP4735026 A1EP 4735026A1EP-4735026-A1

Abstract

The present invention discloses combinatory cancer therapy of immune cells engineered to express an anti-cancer chimeric antigen receptor and klotho polypeptides. The invention discloses various methods to execute the invention as well as compositions and nucleic acid construct that may be used to obtain immune cells that may be used.

Inventors

  • WOLF, IDO
  • GLOBERSON LEVIN, Anat
  • RUBINEK, TAMAR

Assignees

  • Ichilov Tech Ltd.

Dates

Publication Date
20260506
Application Date
20240627

Claims (20)

  1. 1. A therapeutic combination comprising (i) engineered immune cells and (ii) a klotho polypeptide or a vector encoding thereof, for use in treating cancer, wherein the immune cells are engineered to express a chimeric antigen receptor (CAR) that binds specifically to a tumor- associated antigen.
  2. 2. The therapeutic combination for use according to claim 1, wherein the immune cells are characterized by surface expression of the CAR upon administration.
  3. 3. The therapeutic combination according to claim 1 or 2, wherein the use comprises coadministering the engineered immune cells and a klotho polypeptide or a vector encoding thereof in a regimen selected from a sequential administering or a substantially simultaneous administering.
  4. 4. A pharmaceutical composition comprising immune cells engineered to express (i) a chimeric antigen receptor (CAR) that binds specifically to a tumor-associated antigen, and (ii) a klotho polypeptide, and a pharmaceutical acceptable carrier, for use in treating cancer.
  5. 5. The pharmaceutical composition for use according to claim 4, wherein the immune cells express the CAR constitutively.
  6. 6. The pharmaceutical composition for use according to claim 4 or 5, wherein said immune cells comprise a nucleic acid construct comprising a nucleic acid molecule encoding the klotho polypeptide, wherein the nucleic acid molecule is operably linked to an inducible promoter, and wherein the promoter initiates the expression of said klotho polypeptide upon activation of a T cell receptor.
  7. 7. The pharmaceutical composition for use according to claim 6, wherein the immune cells express the klotho polypeptide upon activation of the CAR.
  8. 8. The pharmaceutical composition for use according to claim 4, wherein the immune cells express the CAR and klotho polypeptide constitutively.
  9. 9. A therapeutic combination comprising a population of immune cells engineered to express a CAR that binds specifically to a tumor-associated antigen, and a klotho polypeptide or a vector encoding thereof, for use in improving tumor killing and/or clearance.
  10. 10. A nucleic acid construct comprising a nucleic acid molecule encoding a CAR that binds specifically to a tumor- associated antigen and a nucleic acid molecule encoding a klotho polypeptide, wherein each one of the nucleic acid molecules is operably linked to a promoter or both nucleic acid molecules are operably linked to one promoter.
  11. 11. The nucleic acid construct according to claim 10, wherein both nucleic acid molecules are operably linked to one constitutive promoter.
  12. 12. The nucleic acid construct according to claim 11, further comprising a cleavable nucleic acid molecule or a nucleic acid molecule encoding a self-cleaving peptide between the two nucleic acid molecules, preferably wherein the construct comprises the nucleic acid sequence selected from SEQ ID NO: 66, 37, 38, 116 and 117.
  13. 13. The nucleic acid construct according to claim 10, wherein the nucleic acid molecule encoding the CAR is operably linked to a constitutive promoter and the nucleic acid molecule encoding the klotho polypeptide is operably linked to (i) an inducible promoter, said promoter initiates the expression of klotho polypeptide upon activation of the CAR; or (ii) a constitutive promoter.
  14. 14. A vector comprising the nucleic acid construct according to any one of claims 10 to 13.
  15. 15. A population of immune cells engineered to express (i) a chimeric antigen receptor (CAR) binding specifically to a tumor-associated antigen and (ii) a klotho polypeptide.
  16. 16. The population of immune cells according to claim 15, comprising a nucleic acid construct comprising a nucleic acid molecule encoding the CAR and a nucleic acid construct comprising a nucleic acid molecule encoding the klotho polypeptide.
  17. 17. The population of immune cells according to claim 16, wherein the nucleic acid molecule encoding the CAR is operably linked to a constitutive promoter and the nucleic acid molecule encoding the klotho polypeptide is operably linked to (i) an inducible promoter, said promoter initiates the expression of klotho polypeptide upon activation of the CAR; or (ii) a constitutive promoter.
  18. 18. The population of immune cells according to claim 15, comprising a nucleic acid construct according to any one of claims 10 to 13 or the vector according to claim 14.
  19. 19. A pharmaceutical composition comprising the population of immune cells according to any one of claims 15 to 18, and a pharmaceutically acceptable carrier.
  20. 20. The pharmaceutical composition according to claim 20, for use in treating cancer.

Description

COMBINATION THERAPY USING CAR T CELLS AND KLOTHO FIELD OF THE INVENTION The present invention relates to a combinational cancer therapy using immune cells engineered to express a chimeric antigen receptor (CAR) that binds specifically a tumor- associated antigen, and a klotho polypeptide. The invention relates to different aspects of the therapy such as compositions and methods of providing the treatment. BACKGROUND OF THE INVENTION Therapy based on adoptive cell transfer of Chimeric Antigen Receptor (CAR)- expressing T cells (CAR T cells) has become an innovative highly effective treatment for various hematologic malignancies. CARs are genetically engineered modular single-chain structures composed of an antigen recognition unit and T cell activation moieties in tandem. For the treatment, T cells are harvested from the patient or from a suitable donor, genetically modified ex vivo to express the tumor-directed CAR and re-infused into the patient. The adoptively transferred engineered cells are now able to recognize the respective tumor antigen, and exert a tumor- specific immune response. Despite the success of CAR T cells-based therapy in managing hematologic malignancies, its application toward solid tumors remains challenging, and efforts to design effective CAR T cells for the treatment of several solid tumors, including ovarian and breast cancers, have been unsuccessful (Kosti et al., Front Immunol 2018; 9:1104; Aharon et al, Hum Gene Ther. 2021;32(19-20): 1224-1241). A prerequisite for a success is the ability of CAR T cells to efficiently home and penetrate the desmoplastic stroma surrounding the tumor and accumulate in the tumor microenvironment. Next, cells must gain the ability to expand, persist and mediate cytotoxicity under a hostile milieu largely composed of immunosuppressive modulators. These include myeloid-derived suppressor cells (MDSCs), tumor-associated macrophages (TAMs), and regulatory T cells (Tregs), as well as paracrine signaling by tumor-derived IL-8, which promotes the trafficking of neutrophils and myeloid- derived suppressor cells (MDSCs) into the tumor microenvironment, further dampening anti-tumor immune responses. The klotho gene encodes for a transmembrane protein that can be shed and act as a circulating hormone. Klotho mediates anti-aging activities, and reduced klotho levels are associated with a wide array of aging-related phenomena, including atherosclerosis, dementia, frailty in the elderly and reduced lifespan. These activities are associated with the ability of klotho to reduce inflammation, as demonstrated in the kidney, cardiovascular system, brain and other tissues. Accumulating data indicate klotho as a modulator of the immune system. Klotho knockout mice (kl/kl) show a reduced number of thymocytes, especially CD4 and CD8 double-positive cells, and of both cortical and medullary thymocytes, suggesting a positive role for klotho on the differentiation and proliferation of these cells. Klotho has also been reported to interact with RIG-I and to inhibit RIG-I-induced expression of IL-6 and IL-8 both in vitro and in vivo. It was also shown that kl/kl mice have higher IL-6 levels in the blood as well as in several tissues. Klotho was also suggested as a tumor suppressor in various epithelial malignancies, including breast and ovarian cancers (Wolf et al., Oncogene 2008(27), 7094-7105; Ligumsky et al., Mol Cancer Res 2015, 13(10), 1398- 1407). Its expression in these tumors was reported to be epigenetically silenced, and re-introduction of klotho into cancer cells or treatment with the soluble protein inhibited their viability and aggressiveness in vitro. US 9,987,326 and US 10,555,963, to some of the inventors of the present invention, disclose the use of soluble forms of klotho (e.g., the entire extracellular domain or the KL1 domain), or proteins with at least 80% homology to the amino acid sequence of the soluble forms of klotho, in the treatment of cancer. WO 2020/039425, to some of the inventors of the present invention, discloses compositions and methods for inhibiting tumor growth in subjects in need thereof, utilizing gene transfer vectors, such as viral vectors, comprising a nucleotide sequence encoding a klotho protein operably linked to at least one regulatory sequence directing its expression. WO 2023/218445, to some of the inventors of the present invention, discloses polypeptides derived from klotho KL1 domain exhibiting tumor suppressor activity, and use thereof in treating cancer and inhibiting tumor development. WO 2019111249 and WO 2019111250 to some of the inventors of the present invention, disclose immunological cells comprising dual chimeric antigen receptors and uses thereof in treating cancer. Despite the large number of proposed strategies for cancer treatments, there is still an urgent need for the development of novel efficient therapies for cancer. SUMMARY OF THE INVENTION The present invention is based on unexpected results showing tha