ENGINEERING AND DELIVERY OF THERAPEUTIC COMPOSITIONS OF FRESHLY ISOLATED CELLS

• US 20180028567A1
• Filed: 10/06/2017
• Published: 02/01/2018
• Est. Priority Date: 04/09/2008
• Status: Active Grant

• Current Assignee: Maxcyte, Inc. (Entremed, Incorporated)
• Original Assignee: Maxcyte, Inc. (Entremed, Incorporated)
• Inventors: LI, Linhong, PESHWA, Madhusudan V.
• Granted Patent: US 11,331,344 B2
• Time in Patent Office: Days
• CPC Class Codes: A61K 2035/124 the cells being hematopoietic, bone marrow derived or blood cells
• CPC Class Codes: A61K 39/39558 against tumor tissues, cells, antigens
• CPC Class Codes: A61K 39/4613 Natural-killer cells [NK or NK-T]
• CPC Class Codes: A61K 39/4631 Chimeric Antigen Receptors [CAR]
• CPC Class Codes: A61K 39/464412 CD19 or B4
• CPC Class Codes: A61K 39/464468 Mesothelin [MSLN]
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: C07K 14/7051 T-cell receptor (TcR)-CD3 complex
• CPC Class Codes: C07K 2319/03 containing a transmembrane segment
• CPC Class Codes: C12N 13/00 Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
• CPC Class Codes: C12N 15/85 for animal cells
• CPC Class Codes: C12N 2501/599 with CD designations not provided for elsewhere
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 5/0634 Cells from the blood or the immune system
• CPC Class Codes: C12N 5/0646 Natural killers cells [NK], NKT cells

Abstract

The present invention relates to the transient modification of cells. In particular embodiments, the cells are immune systems, such as PBMC, PBL, T (CD3+ and/or CD8+) and Natural Killer (NK) cells. The modified cells provide a population of cells that express a genetically engineered chimeric receptor which can be administered to a patient therapeutically. The present invention further relates to methods that deliver mRNA coding for the chimeric receptor to unstimulated resting PBMC, PBL, T (CD3+ and/or CD8+) and NK cells and which delivers the mRNA efficiently to the transfected cells and promotes significant target cell killing.

Claims

1-24. -24. (canceled)

25. A modified T cell transiently transfected with mRNA to express a chimeric receptor on its surface, wherein the mRNA comprises a nucleic acid sequence encoding an extracellular domain, a transmembrane domain, a costimulatory signaling region, and a CD3-zeta signaling domain, and wherein the mRNA is transcribed in vitro.

26. The modified T cell of claim 25, wherein the extracellular domain comprises an antigen-binding moiety.

27. The modified T cell of claim 26, wherein the antigen-binding moiety binds to a tumor antigen.

28. The modified T cell of claim 27, wherein the tumor antigen is an antigen associated with a cancer selected from the group consisting of breast cancer, lung cancer, prostate cancer, ovarian cancer, brain cancer, liver cancer, cervical cancer, colon cancer, renal cancer, skin cancer, head &

neck cancer, bone cancer, esophageal cancer, bladder cancer, uterine cancer, lymphatic cancer, stomach cancer, pancreatic cancer, testicular cancer, leukemia, acute lymphocytic leukemia (ALL), acute myelogenous leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), and mantle cell lymphoma (MCL).

29. The modified T cell of claim 25, wherein the chimeric receptor is an anti-mesothelin chimeric receptor.

30. The modified T cell of claim 29, wherein the anti-mesothelin chimeric receptor comprises an anti-mesothelin scFv.

31. The modified T cell of claim 30, wherein the anti-mesothelin scFv is s11.

32. The modified T cell of claim 25, wherein the chimeric receptor comprises an intracellular domain selected from 41BB, CD28, and the cytoplasmic portion of the CD3zeta chain.

33. The modified T cell of claim 29, wherein the anti-mesothelin chimeric receptor comprises an anti-mesothelin scFv, a 4-1 BB intracellular domain, and a CD3zeta domain.

34. A method comprising administering to a subject the modified T cell of claim 25.

35. The method of claim 34, wherein the subject has a hyperproliferative disease.

36. The method of claim 34, wherein the extracellular domain comprises an antigen binding moiety.

37. The method of claim 36, wherein the antigen binding moiety binds to a tumor antigen.

38. The method of claim 35, wherein the hyperproliferative disease is cancer.

39. The method of claim 38, wherein the cancer is selected from the group consisting of breast cancer, lung cancer, prostate cancer, ovarian cancer, brain cancer, liver cancer, cervical cancer, colon cancer, renal cancer, skin cancer, head &

neck cancer, bone cancer, esophageal cancer, bladder cancer, uterine cancer, lymphatic cancer, stomach cancer, pancreatic cancer, testicular cancer, leukemia, acute lymphocytic leukemia (ALL), acute myelogenous leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), and mantle cell lymphoma (MCL).

40. The method of claim 34, wherein the chimeric receptor is an anti-mesothelin chimeric receptor.

41. The method of claim 40, wherein the anti-mesothelin chimeric receptor comprises an anti-mesothelin scFv.

42. The method of claim 41, wherein the anti-mesothelin scFv is ss1.

43. The method of claim 34, wherein the chimeric receptor comprises an intracellular domain selected from 4-1BB, CD28, and the cytoplasmic portion of the CD3zeta chain.

44. The method of claim 40, wherein the anti-mesothelin chimeric receptor comprises an anti-mesothelin scFv, a 4-1 BB intracellular domain, and a CD3zeta domain.