Multimeric IL-15 soluble fusion molecules and methods of making and using same
View Patent ↗The invention provides soluble fusion protein complexes having at least two soluble fusion proteins. The first fusion protein is a biologically active polypeptide covalently linked to an interleukin-15 (IL-15) polypeptide or a functional fragment thereof. The second fusion protein is a second biologically active polypeptide covalently linked to a soluble interleukin-15 receptor alpha (IL-15Rα) polypeptide or a functional fragment thereof. In the complexes of the invention, one or both of the first and second fusion proteins further includes an immunoglobulin Fc domain or a functional fragment thereof; and the first fusion protein binds to the soluble IL-15Rα domain of the second fusion protein to form a soluble fusion protein complex. The invention further provides methods for making and using the complexes of the invention.
1. A method for treating a neoplasia or human immunodeficiency virus (HIV) in a subject in need thereof comprising administering to the subject an effective amount of a pharmaceutical composition comprising an isolated soluble fusion protein complex comprising at least two soluble fusion proteins, wherein
the first fusion protein comprises (a) a first antibody that comprises an anti-CD20 single chain antibody covalently linked to (b) interleukin-15 (IL-15) polypeptide; and
the second fusion protein comprises (c) a second antibody covalently linked to (d) soluble interleukin-15 receptor alpha (IL-15Rα) polypeptide,
wherein one or both of the first and second fusion proteins further comprise an immunoglobulin Fc domain or a functional fragment thereof,
wherein the IL-15 domain of the first fusion protein binds to the soluble IL-15Rα domain of the second fusion protein to form a soluble fusion protein complex,
thereby treating neoplasia or human immunodeficiency virus (HIV) in a subject.
2. The method of claim 1 , wherein the second antibody is directed against a cell surface receptor or ligand.
3. The method of claim 1 , wherein the second antibody is directed against a CD antigen, cytokine or chemokine receptor or ligand, growth factor receptor or ligand, cell adhesion molecule, MHC/MHC-like molecules, Fc receptor, Toll-like receptor, NK receptor, TCR, BCR, positive/negative co-stimulatory receptor or ligand, death receptor or ligand, tumor associated antigen, or virus encoded antigen.
4. The method of claim 1 , wherein the second antibody is directed against CD3, CD4, CD19, CD20, CD21, CD22, CD23, CD25, CD30, CD33, CD38, CD40, CD44, CD51, CD52, CD70, CD74, CD80, CD152, CD147, CD221, EGFR, HER-2/neu, HER-1, HER-3, HER-4, CEA, OX40 ligand, cMet, tissue factor, Nectin-4, PSA, PSMA, EGFL7, FGFR, IL-6 receptor, IGF-1 receptor, GD2, CA-125, EpCam, death receptor 5, MUC1, VEGFR1, VEGFR2, PDGFR, Trail R2, folate receptor, angiopoietin-2, alphavbeta3 integrin receptor, or HLA-DR antigens.
5. The method of claim 1 , wherein the second antibody is directed against HIV, HCV, HBV, CMV, HTLV, HPV, EBV, or RSV or HIV envelope spike, gp120 or gp41.
6. The method of claim 1 , wherein the IL-15 polypeptide comprises an IL-15 polypeptide comprising an N72D mutation (IL-15N72D).
7. The method of claim 1 , wherein
the first fusion protein comprises (a) a first anti-CD20 single chain antibody covalently linked to (b) an IL-15 polypeptide comprising an N72D mutation (IL-15N72D), and wherein
the second fusion protein comprises (c) a second anti-CD20 single chain antibody covalently linked to (d) soluble IL-15 receptor alpha (IL-15Rα) polypeptide.
8. The method of claim 7 , wherein the second fusion protein comprises (c) an anti-CD20 single chain antibody covalently linked to (d) a soluble Il-15receptor alpha sushi-binding domain (IL-15RαSu) fused to an immunoglobulin Fc domain, and wherein the IL-15N72D domain of the first fusion protein binds to the soluble IL-15RαSu domain of the second fusion protein to form a soluble fusion protein complex.
9. The method of claim 8 , wherein the first anti-CD20 single-chain antibody and the second anti-CD20 single-chain antibody each comprise an immunoglobulin light chain variable domain covalently linked to an immunoglobulin heavy chain variable domain by a polypeptide linker sequence.
10. The method of claim 7 , wherein the first fusion protein is encoded by the nucleic acid sequence set forth in SEQ ID NO: 43.
11. The method of claim 7 , wherein the first fusion protein comprises the amino acid sequence set forth in SEQ ID NO: 44.
12. The method of claim 7 , wherein the second fusion protein is encoded by the nucleic acid sequence set forth in SEQ ID NO: 45.
13. The method of claim 7 , wherein the second fusion protein comprises the amino acid sequence set forth in SEQ ID NO: 46.
14. The method of claim 7 wherein the subject is suffering from neoplasia.
15. The method of claim 7 wherein the subject is suffering from human immunodeficiency virus (HIV) in a subject.
16. The method of claim 1 wherein the subject is suffering from neoplasia.
17. The method of claim 1 wherein the subject is suffering from human immunodeficiency virus (HIV) in a subject.
18. The method of claim 1 wherein the second fusion protein comprises (c) a second anti-CD20 single chain antibody covalently linked to (d) soluble interleukin-15 receptor alpha (IL-15Rα) polypeptide.