Bivalent IL-2 fusion toxins

Patent metadata

Field	Value
Publication number	US-9764006-B2
Application number	US-201314650699-A
Country	US
Kind code	B2
Filing date	Dec 9, 2013
Priority date	Dec 10, 2012
Publication date	Sep 19, 2017
Grant date	Sep 19, 2017

Abstract

IL-2 fusion toxins, e.g., bivalent-IL2 fusion toxins, and methods of use thereof.

First claim

What is claimed is:

1. A bivalent IL-2 fusion toxin comprising:
a first part comprising a cytotoxic protein, and
a second part comprising at least two Interleukin 2 (IL-2) sequences comprising amino acids 21-153 of SEQ ID NO:1, wherein the cytotoxic protein comprises diphtheria toxin, Pseudomonas exotoxin , or cytotoxic portions or variants thereof.

2. The fusion toxin of claim 1 , wherein the cytotoxic protein comprises diphtheria toxin, or cytotoxic portions or variants thereof.

3. The fusion toxin of claim 1 , further comprising a linker between the first and second parts.

4. The fusion toxin of claim 1 , wherein fusion toxin comprises a linker between the two IL-2 sequences.

5. A codon-optimized nucleic acid molecule optimized for expression in a methylotropic yeast encoding the fusion toxin of claim 1 .

6. A nucleic acid encoding the fusion toxin of claim 1 .

7. A vector comprising the nucleic acid molecule of claim 6 .

8. A host cell expressing the nucleic acid molecule of claim 5 .

9. The host cell of claim 8 , wherein the host cell is a methylotropic yeast.

10. The host cell of claim 8 , wherein the host cell is a cell of the species Pichia Pastoris.

11. A pharmaceutical composition comprising the fusion toxin of claim 1 , and a physiologically acceptable carrier.

12. A method of treating a subject who has a cancer, the method comprising administering to the subject a therapeutically effective amount of the fusion toxin of claim 1 .

13. The method of claim 12 , wherein the cancer comprises cancer cells that express CD25.

14. The method of claim 12 , further comprising administering an immunotherapy to the subject.

15. The method of claim 14 , wherein the immunotherapy comprises administration of one or more of: dendritic cells or peptides with adjuvant; DNA-based vaccines; cytokines; cyclophosphamide; anti-interleukin-2R immunotoxins; antibodies; virus-based vaccines ; formulations of Toll-like Receptor or RIG-I-like receptor ligands; or adoptive T cell therapy or other cell therapy.

16. The method of claim 12 , wherein the cancer is selected from the group consisting of B-cell neoplasms, acute nonlymphocytic leukemias, neuroblastomas, tumor infiltrating lymphocytes, and cutaneous T cell lymphoma.

17. A method of depleting CD25-expressing regulatory T cells in a subject, the method comprising administering to the subject an effective amount of the fusion toxin of claim 1 .

18. The method of claim 17 , wherein the subject has cancer, or is an experimental model of autoimmune disease or transplant rejection.

19. A method of producing a bivalent IL-2 fusion toxin, the method comprising:
expressing a codon-optimized nucleic acid molecule encoding the fusion toxin of claim 1 in a methylotropic yeast; and
substantially purifying the fusion toxin,
thereby producing the the fusion toxin.

20. The method of claim 19 , wherein the methylotropic yeast is of the species Pichia Pastoris.

Assignees

• Massachusetts Gen Hospital

Inventors

• Wang Zhirui
• Huang Christene A
• Sachs David H

Classifications

• A61K38/00

  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 title

• A61K38/45 Primary

  Transferases (2) · CPC title

• A61K38/2013

  IL-2 · CPC title

• C07K2319/55

  containing a fusion with a toxin, e.g. diphteria toxin · CPC title

• C12N15/63

  Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression · CPC title