Methods and Compositions for Promoting Immune Cell Function

• US 20200131239A1
• Filed: 06/13/2017
• Published: 04/30/2020
• Est. Priority Date: 06/13/2016
• Status: Active Grant

• Current Assignee: Torque Therapeutics, Inc. (Repertoire Immune Medicines, Inc.)
• Original Assignee: Torque Therapeutics, Inc. (Repertoire Immune Medicines, Inc.)
• Inventors: PUCCI, Ferdinando, ANDRESEN, Thomas Lars, JONES, Douglas Scott, NIELSEN, Ulrik, RAKESTRAW, James Andrew
• Granted Patent: US 11,472,856 B2
• Time in Patent Office: Days
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 38/1793 for cytokines; for lymphokines; for interferons
• CPC Class Codes: A61K 38/2086 IL-13 to IL-16
• CPC Class Codes: A61K 47/68 the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
• CPC Class Codes: A61K 47/6901 Conjugates being cells, cell fragments, viruses, ghosts, red blood cells or viral vectors
• CPC Class Codes: A61P 11/06 Antiasthmatics
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 37/08 Antiallergic agents antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: C07K 14/54 Interleukins [IL]
• CPC Class Codes: C07K 14/56 IFN-alpha
• CPC Class Codes: C07K 14/565 IFN-beta
• CPC Class Codes: C07K 14/70514 CD4
• CPC Class Codes: C07K 14/70517 CD8
• CPC Class Codes: C07K 14/70553 Integrin beta2-subunit-containing molecules, e.g. CD11, CD18
• CPC Class Codes: C07K 14/70589 CD45
• CPC Class Codes: C07K 2317/52 Constant or Fc region; Isotype
• CPC Class Codes: C07K 2319/30 Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto

Abstract

The present disclosure features, at least in part, methods for conserving cell function, e.g., immune cell function, e.g., after one or more cycles of freezing and/or thawing the nucleated cell. In embodiments, the methods comprise contacting an immune cell with a protein nanoparticle comprising an IL-15 complex.

Claims

1. A composition comprising a nucleated cell and a nanoparticle that comprises at least one conservation agent, wherein:

(a) at least part of the composition is frozen,

(b) the conservation agent comprises a cytokine molecule or a costimulatory molecule, and

(c) wherein;

(i) the cytokine molecule is chosen from IL15, IL2, IL6, IL7, IL12, IL17, IL18, IL21, IL-23, IL-4, IL1alpha, IL1beta, IL-5, IFNgamma, TNFa, IFNalpha, IFNbeta, GM-CSF, or GCSF; and

/or

(ii) the costimulatory molecule is chosen from an, OX40, CD28, GITR, VISTA, CD40, CD3, or an agonist of CD137.

2-4. -4. (canceled)

5. A composition comprising a nucleated cell and a nanoparticle that comprises at least one conservation agent, wherein:

(a) at least part of the composition is frozen; and

(b) the conservation agent comprises an IL-15 molecule complexed with a polypeptide comprising;

a first domain comprising the amino acid sequence of SEQ ID NO;

7 or SEQ ID NO;

8, or an amino acid sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity thereto, and having IL-15 binding activity; and

optionally, a second, heterologous domain comprising an antibody molecule or an Fc domain.

6. The composition of claim 5, wherein upon thawing of the composition, the conservation agent improves one or more of viability, proliferation, cytotoxic activity or activation of the nucleated cell compared to a nucleated cell in the absence of the nanoparticle.

18. The composition of claim 5, wherein the IL-15 molecule is wild-type IL-15 or a polypeptide of SEQ ID NO:

6 or an amino acid sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity thereto, which has IL-15Ra binding activity.

19. The composition of claim 5, wherein the polypeptide comprising the IL-15 receptor or fragment thereof comprises a polypeptide of SEQ ID NO:

5 or an amino acid sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity thereto, which has IL-15 binding activity.

25. The composition according to claim 5, wherein the nucleated cell is an immune effector cell or a hematopoietic stem cell, selected from a lymphocyte, T cell, B cell, or Natural Killer (NK) cell.

38. The composition according to claim 5, comprising a second conservation agent which comprises a cytokine molecule.

39. The composition of claim 38, wherein the cytokine molecule comprises IL2, IL6, IL7, IL12, IL15, IL17, IL18, IL21, IL-23, IL-4, IL1alpha, IL1beta, IL-5, IFNgamma, TNFa, IFNalpha, IFNbeta, GM-CSF, or GCSF, or a variant thereof.

41. The composition according to claim 5, wherein the nanoparticle comprises an antibody molecule to CD45, CD11a (integrin alpha-L), CD18 (integrin beta-2), CD11b, CD11c, CD25, CD8, or CD4.

43. The composition according to claim 5, wherein the nanoparticle comprises a nanogel that is cross linked by a reversible linker that is sensitive to redox, pH, or enzymes.

7-17. -17. (canceled)

20-24. -24. (canceled)

26-37. -37. (canceled)

40. (canceled)

42. (canceled)

44-46. -46. (canceled)

47. A method for enhancing one or more of the viability, proliferation, cytotoxic activity or activation of a nucleated cell, comprising:

contacting the nucleated cell with a nanoparticle that comprises at least one conservation agent;

subjecting the contacted nucleated cell to at least one freeze thaw cycle, wherein the conservation agent comprises an IL-15 molecule complexed with a polypeptide comprising;

a first domain comprising the amino acid sequence of SEQ ID NO;

7 or SEQ ID NO;

8, or an amino acid sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity thereto, and having IL-15 binding activity.

49. The method of claim 47, wherein the temperature of the composition is reduced to less than 0 degrees centigrade or less than negative 10 degrees centigrade.

56. The method of claim 47, wherein the conservation agent comprises

a second, heterologous domain comprising an antibody molecule or an Fc domain.

62. The method of claim 47, wherein the nucleated cell is an immune effector cell or a hematopoietic stem cell.

63. The method of claim 62, wherein the nucleated cell is a lymphocyte, T cell, B cell, or NK cell.

69. The method of claim 47, wherein the nanoparticle comprises a cationic block co-polymer comprising PEG and a poly-lysine.

75. The method of claim 47, wherein the composition comprises an antibody molecule to CD45, CD11a (integrin alpha-L), CD18 (integrin beta-2), CD11b, CD11c, CD25, CD8, or CD4.

76. The method of claim 47, wherein the nanogel is greater than 80% protein by weight, and/or wherein the protein is cross-linked by a cross-linking molecule that upon degradation releases the proteins in a native form.

78. The method of claim 47, wherein the nanoparticle comprises

a polypeptide of SEQ ID NO:

6 or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity thereto which has IL-15Ra binding activity;

a polypeptide of SEQ ID NO;

5 or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity thereto, which has IL-15 binding activity;

a cationic block co-polymer comprising PEG having a molecular weight of about 1-10, 2-8, 4-6, or about 5 kD and a poly-lysine having an average length of 10-50, 20-40, or about 30 amino acids; and

optionally, an antibody molecule to CD45.

48. (canceled)

50-55. -55. (canceled)

57-61. -61. (canceled)

64-68. -68. (canceled)

70-74. -74. (canceled)

77. (canceled)

79-104. -104. (canceled)