MUTANT INTERLEUKIN-2 POLYPEPTIDES

• US 20190322765A1
• Filed: 06/17/2019
• Published: 10/24/2019
• Est. Priority Date: 02/10/2011
• Status: Abandoned Application

• Current Assignee: Roche Glycart AG (Roche Holding AG)
• Original Assignee: Roche Glycart AG (Roche Holding AG)
• Inventors: Ast, Oliver, Bruenker, Peter, Freimoser-Grundschober, Anne, Herter, Sylvia, Hofer, Thomas U., Hosse, Ralf, Klein, Christian, Moessner, Ekkehard, Nicolini, Valeria G., Umana, Pablo
• Application Number:
  US16/443,306
• Publication Number:
  US 20190322765A1
• Time in Patent Office: Days
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 38/2013 IL-2
• CPC Class Codes: A61K 39/39558 against tumor tissues, cells, antigens
• CPC Class Codes: A61K 47/6813 the drug being a peptidic cytokine, e.g. an interleukin or interferon
• CPC Class Codes: A61K 47/6849 the antibody targeting a receptor, a cell surface antigen or a cell surface determinant
• CPC Class Codes: A61K 47/6871 the antibody targeting an enzyme
• CPC Class Codes: A61K 47/6891 Pre-targeting systems involving an antibody for targeting specific cells
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: C07K 14/55 IL-2
• CPC Class Codes: C07K 16/28 against receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: C07K 16/30 from tumour cells
• CPC Class Codes: C07K 16/40 against enzymes
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C12N 15/09 Recombinant DNA-technology
• CPC Class Codes: C12N 15/62 DNA sequences coding for fusion proteins

Abstract

The present invention generally relates to mutant interleukin-2 polypeptides that exhibit reduced affinity to the α-subunit of the IL-2 receptor, for use as immunotherapeutic agents. In addition, the invention relates to immunoconjugates comprising said mutant IL-2 polypeptides, polynucleotide molecules encoding the mutant IL-2 polypeptides or immunoconjugates, and vectors and host cells comprising such polynucleotide molecules. The invention further relates to methods for producing the mutant IL-2 polypeptides or immunoconjugates, pharmaceutical compositions comprising the same, and uses thereof.

Claims

1. A mutant interleukin-2 (IL-2) polypeptide comprising at a first amino acid mutation that abolishes or reduces affinity of the mutant IL-2 polypeptide to the high-affinity IL-2 receptor and preserves affinity of the mutant IL-2 polypeptide to the intermediate-affinity IL-2 receptor, each compared to a wild-type IL-2 polypeptide, characterized in that said first amino acid mutation is at a position corresponding to residue 72 of human IL-2.

2. The mutant interleukin-2 polypeptide of claim 1, wherein said first amino acid mutation is an amino acid substitution, selected from the group of L72G, L72A, L72S, L72T, L72Q, L72E, L72N, L72D, L72R, and L72K.

3. The mutant interleukin-2 polypeptide of claim 1 or 2, comprising a second amino acid mutation that abolishes or reduces affinity of the mutant IL-2 polypeptide to the high-affinity IL-2 receptor and preserves affinity of the mutant IL-2 polypeptide to the intermediate-affinity IL-2 receptor, each compared to a wild-type IL-2 polypeptide.

4. The mutant interleukin-2 polypeptide of claim 3, wherein said second amino acid mutation is at a position selected from the positions corresponding to residue 35, 38, 42, 43, and 45 of human IL-2.

5. The mutant interleukin-2 polypeptide of claim 3 or 4, wherein said second amino acid mutation is at a position corresponding to residue 42 of human IL-2.

6. The mutant interleukin-2 polypeptide of claim 5, wherein said second amino acid mutation is an amino acid substitution, selected from the group of F42A, F42G, F42S, F42T, F42Q, F42E, F42N, F42D, F42R, and F42K.

7. The mutant interleukin-2 polypeptide of any one of claims 3 to 6, comprising a third amino acid mutation that abolishes or reduces affinity of the mutant IL-2 polypeptide to the high-affinity IL-2 receptor and preserves affinity of the mutant IL-2 polypeptide to the intermediate-affinity IL-2 receptor, each compared to a wild-type IL-2 polypeptide.

8. The mutant interleukin-2 polypeptide of any one of claims 1 to 7, comprising three amino acid mutations that abolish or reduce affinity of the mutant IL-2 polypeptide to the high-affinity IL-2 receptor and preserve affinity of the mutant IL-2 polypeptide to the intermediate-affinity IL-2 receptor, each compared to a wild-type IL-2 polypeptide, wherein said three amino acid mutations are at positions corresponding to residue 42, 45, and 72 of human IL-2.

9. The mutant interleukin-2 polypeptide of claim 8, wherein said three amino acid mutations are amino acid substitutions selected from the group of F42A, F42G, F42S, F42T, F42Q, F42E, F42N, F42D, F42R, F42K, Y45A, Y45G, Y45S, Y45T, Y45Q, Y45E, Y45N, Y45D, Y45R, Y45K, L72G, L72A, L72S, L72T, L72Q, L72E, L72N, L72D, L72R, and L72K.

10. The mutant interleukin-2 polypeptide of any one of claims 1 to 9, further comprising an amino acid mutation which eliminates the O-glycosylation site of IL-2 at a position corresponding to residue 3 of human IL-2.

11. The mutant interleukin-2 polypeptide of any one of claims 1 to 10, wherein said mutant IL-2 polypeptide is linked to a non-IL-2 moiety.

12. The mutant interleukin-2 polypeptide of any one of claims 1 to 11, wherein said mutant IL-2 polypeptide is linked to a first and a second non-IL-2 moiety.

13. The mutant interleukin-2 polypeptide of claim 12, wherein said mutant IL-2 polypeptide shares a carboxy-terminal peptide bond with said first non-IL-2 moiety and an amino-terminal peptide bond with said second non-IL-2 moiety.

14. The mutant interleukin-2 polypeptide of any one of claims 11 to 13, wherein said non-IL-2 moiety is an antigen binding moiety.

15. An immunoconjugate comprising a mutant IL-2 polypeptide according to any one of claims 1 to 10 and an antigen binding moiety.

16. The immunoconjugate of claim 15, wherein said mutant IL-2 polypeptide shares an amino- or carboxy-terminal peptide bond with said antigen binding moiety.

17. The immunoconjugate of claim 15 or 16, wherein said immunoconjugate comprises as first and a second antigen binding moiety.

18. The immunoconjugate of claim 17, wherein said mutant IL-2 polypeptide shares an amino- or carboxy-terminal peptide bond with said first antigen binding moiety and said second antigen binding moiety shares an amino- or carboxy-terminal peptide bond with either i) said mutant IL-2 polypeptide or ii) said first antigen binding moiety.

19. The mutant interleukin-2 polypeptide of claim 14 or the immunoconjugate of any one of claims 15 to 18, wherein said antigen binding moiety is an antibody or an antibody fragment.

20. The mutant interleukin-2 polypeptide of claim 14 or the immunoconjugate of any one of claims 15 to 18, wherein said antigen binding moiety is selected from a Fab molecule and a scFv molecule.

21. The mutant interleukin-2 polypeptide of claim 14 or the immunoconjugate of any one of claims 15 to 18, wherein said antigen binding moiety is an immunoglobulin molecule, particularly an IgG molecule.

22. The mutant interleukin-2 polypeptide of claim 14 or the immunoconjugate of any one of claims 15 to 21, wherein said antigen binding moiety is directed to an antigen presented on a tumor cell or in a tumor cell environment.

23. The mutant interleukin-2 polypeptide or immunoconjugate of claim 22, wherein said antigen is selected from the group of Fibroblast Activation Protein (FAP), the A1 domain of Tenascin-C (TNC A1), the A2 domain of Tenascin-C (TNC A2), the Extra Domain B of Fibronectin (EDB), Carcinoembryonic Antigen (CEA) and the Melanoma-associated Chondroitin Sulfate Proteoglycan (MCSP).

24. An isolated polynucleotide encoding the mutant IL-2 polypeptide or immunoconjugate of any one of claims 1 to 23.

25. An expression vector comprising the polynucleotide of claim 24.

26. A host cell comprising the polynucleotide of claim 24 or the expression vector of claim 25.

27. A method of producing a mutant IL-2 polypeptide or an immunoconjugate thereof, comprising culturing the host cell of claim 26 under conditions suitable for the expression of the mutant IL-2 polypeptide or the immunoconjugate.

28. A mutant IL-2 polypeptide or immunoconjugate produced by the method of claim 27.

29. A pharmaceutical composition comprising the mutant IL-2 polypeptide or immunoconjugate of any one of claims 1 to 23 or 28 and a pharmaceutically acceptable carrier.

30. The mutant IL-2 polypeptide or immunoconjugate of any one of claims 1 to 23 or 28 for use in the treatment of a disease in an individual in need thereof.

31. The mutant IL-2 polypeptide or immunoconjugate of claim 30, wherein said disease is cancer.

32. Use of the mutant IL-2 polypeptide or immunoconjugate of any one of claims 1 to 23 or 28 for manufacture of a medicament for treating a disease in an individual in need thereof.

33. A method of treating disease in an individual, comprising administering to said individual a therapeutically effective amount of a composition comprising the mutant IL-2 polypeptide or immunoconjugate of any one of claims 1 to 23 or 28 in a pharmaceutically acceptable form.

34. The method of claim 33, wherein said disease is cancer.

35. A method of stimulating the immune system of an individual, comprising administering to said individual a effective amount of a composition comprising the mutant IL-2 polypeptide or immunoconjugate of any one of claims 1 to 23 or 28 in a pharmaceutically acceptable form.

36. The invention as described hereinbefore.