Fusokines Involving Cytokines with Strongly Reduced Receptor Binding Affinities

• US 20180155404A1
• Filed: 07/03/2014
• Published: 06/07/2018
• Est. Priority Date: 07/18/2013
• Status: Active Grant

• Current Assignee: Centre Hospitalier Régional Universitaire de Montpellier, Centre National De La Recherche Scientifique, Universiteit Gent Vzw, VIB VZW
• Original Assignee: Centre Hospitalier Régional Universitaire de Montpellier, Centre National De La Recherche Scientifique, Universiteit Gent Vzw, Universite Montpellier II, VIB VZW
• Inventors: TAVERNIER, Jan, BULTNICK, Jennyfer, GERLO, Sarah, UZ, Gilles, PAUL, Franciane, BORDAT, Yann
• Granted Patent: US 10,640,542 B2
• Time in Patent Office: Days
• CPC Class Codes: A61K 38/195 Chemokines, e.g. RANTES
• CPC Class Codes: A61K 38/2006 IL-1
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: C07K 14/52 Cytokines; Lymphokines; Interferons
• CPC Class Codes: C07K 14/521 Chemokines
• CPC Class Codes: C07K 14/523 Beta-chemokines, e.g. RANTES, I-309/TCA-3, MIP-1alpha, MIP-1beta/ACT-2/LD78/SCIF, MCP-1/MCAF, MCP-2, MCP-3, LDCF-1, LDCF-2
• CPC Class Codes: C07K 14/525 Tumour necrosis factor [TNF]
• CPC Class Codes: C07K 14/545 IL-1
• CPC Class Codes: C07K 14/56 IFN-alpha
• CPC Class Codes: C07K 14/5759 Products of obesity genes, e.g. leptin, obese (OB), tub, fat
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/33 fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies
• CPC Class Codes: C07K 2319/74 containing a fusion for binding to a cell surface receptor

Abstract

The present invention relates to a fusion protein comprising at least two cytokines, of which at least one is a modified cytokine with a strongly reduced binding affinity to its receptor, or to one of its receptors. Preferably, both cytokines are connected by a linker, preferably a GGS linker. The invention relates further to said fusion protein for use in treatment of diseases.

Claims

1. A composition comprising a fusion protein comprising at least two cytokines, wherein:

at least one cytokine comprises a mutation that strongly reduces binding activity to its receptor, and

at least one cytokine is a wild-type cytokine which provides cell-specific targeting that restores activity of the mutant cytokine on the targeted cells.

2. The fusion protein according to claim 1, further comprising a GGS linker.

3. The composition according to claim 1, wherein said cytokines are XCL1 and IFNα

2.

4. The composition according to claim 1, wherein said cytokines are CCL20 and IL-1β

.

5. The composition according to claim 1, wherein said cytokines are TNF and leptin.

9. The composition according to claim 3, wherein the IFNα

2 comprises the mutation.

10. The composition according to claim 9, wherein the XCL1 is not mutated.

11. The composition according to claim 4, wherein the IL-1β

comprises the mutation.

12. The composition of claim 11, wherein the IL-1β

mutation is at a position selected from 120, 131, 146, 148, and 209.

13. The composition of claim 12, wherein the IL-1β

mutation is selected from R120G, Q131G, H146A, Q148G, and K209A.

14. The composition according to claim 11, wherein the CCL20 is not mutated.

15. The composition according to claim 5, wherein the leptin comprises the mutation.

16. The composition of claim 15, wherein the leptin mutation is at position 86.

17. The composition of claim 16, wherein the leptin mutation is selected from L86S and L86N.

18. The composition according to claim 15, wherein the TNF is not mutated.

6-8. -8. (canceled)

19. A method of stimulating an immune response in a cell, comprising contacting the cell with a composition comprising a fusion protein comprising at least two cytokines, wherein:

at least one of the cytokines has a mutation which provides strongly reduced affinity for its receptor as compared to wild type cytokine.

20. The method of claim 19, wherein NFκ

B is activated.

21. The method of claim 20, wherein the mutated cytokine is a human IL-1β

mutant comprising a mutation selected from R120G, Q131G, H146A, Q148G, and K209A.

22. The method of claim 21, wherein one of the cytokines is wild type CCL20.