Chimeric antigen receptor-expressing T cells as anti-cancer therapeutics

Patent metadata

Field	Value
Publication number	US-12150981-B2
Application number	US-202117387892-A
Country	US
Kind code	B2
Filing date	Jul 28, 2021
Priority date	Dec 20, 2012
Publication date	Nov 26, 2024
Grant date	Nov 26, 2024

Abstract

Cytotoxic lymphocytes expressing chimeric antigen receptors (CAR) that target and bind small conjugate molecules (SCM) are disclosed, as well as methods of using the cells and the SCMs in the treatment of cancer.

First claim

What is claimed is:

1. A method of killing folate receptor-positive (FR+) cancer cells, the method comprising:
administering a fluorescein isothiocyanate-folate (FITC-folate) conjugate to a subject that has FR+ cancer cells; and
contacting the FR+ cancer cells, the folate receptors of which are bound by fluorescein isothiocyanate-folate (FITC-folate) conjugates, with an effective amount of cytotoxic lymphocytes expressing a chimeric antigen receptor (CAR) that specifically binds FITC.

2. The method of claim 1 , wherein the CAR is a fusion protein comprising a recognition region, a transmembrane domain, a co-stimulation domain, and an activation signaling domain, and wherein the recognition region of the CAR has binding specificity for FITC.

3. The method of claim 2 , wherein the recognition region of the CAR is a single chain fragment variable (scFv) region of an anti-FITC antibody.

4. The method of claim 1 , wherein the FITC-folate conjugates comprise a polyethylene glycol-12 ((PEG) 12 ) linker, and the FITC and the folate are conjugated via the linker (FITC-(PEG) 12 -folate conjugates).

5. The method of claim 1 , wherein the FITC-folate conjugates comprise an ethylenediamine linker, and the FITC and folate are conjugated via the linker (FITC-ethylenediamine-folate conjugates).

6. The method of claim 1 , wherein a plurality of the FITC-folate conjugates are not endocytosed by the folate receptors of the FR+ cancer cells.

7. The method of claim 1 , wherein induction of cytokine storm is reduced.

8. The method of claim 1 , wherein the method comprises transfecting cytotoxic lymphocytes with a vector encoding the CAR that specifically binds FITC prior to contacting the FR+ cancer cells with the cytotoxic lymphocytes expressing the CAR that specifically binds FITC.

9. The method of claim 8 , wherein the vector is a lentiviral vector.

10. The method of claim 1 , wherein the method causes lysis of at least about 18% of the FR+ cancer cells.

11. The method of claim 1 , wherein the method causes lysis of at least about 29% of the FR+ cancer cells.

12. The method of claim 1 , wherein the method causes lysis of at least about 51% of the FR+ cancer cells.

13. The method of claim 1 , wherein the method causes lysis of about 18% of the FR+ cancer cells.

14. The method of claim 1 , wherein the method causes lysis of about 29% of the FR+ cancer cells.

15. The method of claim 1 , wherein the method causes lysis of about 51% of the FR+ cancer cells.

16. The method of claim 1 , wherein the cytotoxic lymphocytes are cytotoxic T lymphocytes.

17. The method of claim 16 , wherein the CAR is a fusion protein comprising a recognition region, a transmembrane domain, a co-stimulation domain, and an activation signaling domain, and wherein the recognition region of the CAR has binding specificity for FITC.

18. The method of claim 17 , wherein the recognition region of the CAR is a scFv region of an anti-FITC antibody.

19. The method of claim 16 , wherein the FITC-folate conjugates comprise a polyethylene glycol-12 ((PEG) 12 ) linker, and the FITC and the folate are conjugated via the linker (FITC-(PEG) 12 -folate conjugates).

20. The method of claim 16 , wherein the FITC-folate conjugates comprise an ethylenediamine linker, and the FITC and the folate are conjugated via the linker (FITC-ethylenediamine-folate conjugates).

21. The method of claim 16 , wherein a plurality of the FITC-folate conjugates are not endocytosed by the folate receptors of the FR+ cancer cells.

22. The method of claim 16 , wherein the induction of cytokine storm is reduced.

23. The method of claim 16 , wherein the method further comprises transfecting cytotoxic lymphocytes with a vector encoding the CAR that specifically binds FITC prior to contacting the FR+ cancer cells with the cytotoxic lymphocytes expressing the CAR that specifically binds FITC.

24. The method of claim 23 , wherein the vector is a lentiviral vector.

25. The method of claim 16 , wherein the method causes lysis of at least about 18% of the FR+ cancer cells.

26. The method of claim 16 , wherein the method causes lysis of at least about 29% of the FR+ cancer cells.

27. The method of claim 16 , wherein the method causes lysis of at least about 51% of the FR+ cancer cells.

28. The method of claim 16 , wherein the method causes lysis of about 18% of the FR+ cancer cells.

29. The method of claim 16 , wherein the method causes lysis of about 29% of the FR+ cancer cells.

30. The method of claim 16 , wherein the method causes lysis of about 51% of the FR+ cancer cells.

31. A method of killing folate receptor-positive (FR+) cancer cells in a patient, which method comprises:
administering to the patient, to whom has already been administered a fluorescein isothiocyanate-folate (FITC-folate) conjugate, an effective amount of cytotoxic lymphocytes expressing a chimeric antigen receptor (CAR) that specifically binds FITC,
wherein the cytotoxic lymphocytes bind FITC-folate conjugate bound to folate receptors on the FR+ cancer cells in the patient.

Assignees

• Purdue Research Foundation

Inventors

• Low Philip S
• Chu Haiyan
• Lee Yong Gu

Classifications

• A61K40/31

  Chimeric antigen receptors [CAR] · CPC title

• A61K40/11

  T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells · CPC title

• A61K40/4202

  Receptors, cell surface antigens or cell surface determinants · CPC title

• A61K2239/13

  Antibody-based · CPC title

• A61K2039/585

  wherein the target is cancer · CPC title