Il-2 variants for the treatment of autoimmune diseases

Abstract

This disclosure provides for a method for treating a condition by administering to a subject in need thereof a therapeutically-effective amount of a compound that comprises an IL-2 receptor-binding moiety. The methods described in the present disclosure provide enhanced pharmacokinetic profiles. The disclosure also provides methods for treating autoimmune disease comprising administering a therapeutically-effective amount of a fusion protein comprising an IL-2 variant protein.

We claim: 
     
         1 . A method for treating an autoimmune disease, the method comprising administering to a subject in need thereof at least two doses of a pharmaceutical composition comprising a therapeutically-effective amount of a fusion protein comprising:
 a. a human IL-2 variant protein domain comprising a substitution selected from the group consisting of T3A and C125S relative to the amino acid sequence of SEQ ID NO: 2, and a substitution selected from the group consisting of D20H, N88I, N88G, N88R, Q126L, and Q126F relative to the amino acid sequence of SEQ ID NO: 2;   b. a peptide linker domain; and   c. an IgG Fc protein domain,   
       wherein each domain has an amino-terminus (N-terminus) and a carboxy-terminus (C-terminus); and wherein the fusion protein is configured so that the C-terminus of the human IL-2 variant protein domain is fused through a peptide bond to the N-terminus of the peptide linker domain, and the N-terminus of the IgG Fc protein domain is fused through a peptide bond to the C-terminus of the peptide linker domain, 
       wherein the composition is administered to the subject at a dosing frequency from once every week to once every month and at a dose of 20 μg/kg to 2 mg/kg, 
       wherein administration of the pharmaceutical composition to the subject increases the level of regulatory T cells in the subject by at least 1.76-fold five days after administration relative to the level of regulatory T cells in the subject before treatment with the pharmaceutical composition, and wherein the autoimmune disease is selected from the group consisting of Psoriasis, Ulcerative Colitis, Crohn's disease, Pemphigus Vulgaris, Type 1 Diabetes, Systemic Lupus Erythematosus, Graft-versus-Host Disease, Autoimmune Vasculitis, Multiple Sclerosis, Amytrophic Lateral Sclerosis, Alopecia Areata, Uveitis, Duchenne Muscular Dystrophy, Scleroderma, and Neuromyelitis Optica. 
     
     
         2 . The method of  claim 1 , wherein the dosing frequency ranges from once every week to once every 2 weeks. 
     
     
         3 . The method of  claim 1 , wherein the human IL-2 variant protein domain comprises the N88R substitution relative to the amino acid sequence of SEQ ID NO: 2. 
     
     
         4 . The method of  claim 1 , wherein the human IL-2 variant protein domain comprises the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 3. 
     
     
         5 . The method of  claim 1 , wherein the peptide linker domain comprises the amino acid sequence of SEQ ID NO: 6. 
     
     
         6 . The method of  claim 1 , wherein the IgG Fc protein domain comprises the amino acid sequence of SEQ ID NO: 7. 
     
     
         7 . The method of  claim 1 , wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 4 or SEQ ID NO: 5. 
     
     
         8 . The method of  claim 1 , wherein the pharmaceutical composition is administered to the subject by subcutaneous administration. 
     
     
         9 . A method for increasing proliferation of regulatory T cells in a subject in need thereof, the method comprising administering to the subject at least two doses of a pharmaceutical composition comprising a therapeutically effective amount of a fusion protein comprising:
 a. a human IL-2 variant protein domain comprising a substitution selected from the group consisting of T3A and C125S relative to the amino acid sequence of SEQ ID NO: 2, and a substitution selected from the group consisting of D20H, N88I, N88G, N88R, Q126L, and Q126F relative to the amino acid sequence of SEQ ID NO: 2;   b. a peptide linker domain; and   c. an IgG Fc protein domain,   
       wherein each domain has an amino-terminus (N-terminus) and a carboxy terminus (C-terminus); and wherein the fusion protein is configured so that the C-terminus of the human IL-2 variant protein domain is fused through a peptide bond to the N-terminus of the peptide linker domain, and the N-terminus of the IgG Fc protein domain is fused through a peptide bond to the C-terminus of the peptide linker domain, 
       wherein the composition is administered to the subject at a dosing frequency from once every week to once every month and at a dose of 20 μg/kg to 2 mg/kg, and 
       wherein administration of the pharmaceutical composition to the subject increases the level of regulatory T cells in the subject by at least 1.76-fold five days after administration relative to the level of regulatory T cells in the subject before treatment with the pharmaceutical composition. 
     
     
         10 . The method of  claim 9 , wherein administration of the pharmaceutical composition to the subject increases the level of regulatory T cells by at least 2-fold relative to the level of regulatory T cells in the subject before treatment with the pharmaceutical composition. 
     
     
         11 . The method of  claim 9 , wherein administration of the pharmaceutical composition to the subject does not increase the proliferation of conventional T cells or CD8+ T cells. 
     
     
         12 . The method of  claim 9 , wherein the human IL-2 variant protein domain comprises the N88R substitution relative to the amino acid sequence of SEQ ID NO: 2. 
     
     
         13 . The method of  claim 9 , wherein the peptide linker domain comprises the amino acid sequence of SEQ ID NO: 6. 
     
     
         14 . The method of  claim 9 , wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 4 or SEQ ID NO: 5. 
     
     
         15 . A method for maintaining the ratio of regulatory T cells (Treg) to conventional T cells (Tconv) at a level of at least 0.2 in a subject, the method comprising administering to the subject at least two doses of a pharmaceutical composition comprising a therapeutically effective amount of a fusion protein comprising:
 a. a human IL-2 variant protein domain comprising a substitution selected from the group consisting of T3A and C125S relative to the amino acid sequence of SEQ ID NO: 2, and a substitution selected from the group consisting of D20H, N88I, N88G, N88R, Q126L, and Q126F relative to the amino acid sequence of SEQ ID NO: 2;   b. a peptide linker domain; and   c. an IgG Fc protein domain,   
       wherein each domain has an amino-terminus (N-terminus) and a carboxy terminus (C-terminus); and wherein the fusion protein is configured so that the C-terminus of the human IL-2 variant protein domain is fused through a peptide bond to the N-terminus of the peptide linker domain, and the N-terminus of the IgG Fc protein domain is fused through a peptide bond to the C-terminus of the peptide linker domain, 
       wherein the composition is administered to the subject at a dosing frequency from once every week to once every month and at a dose of 20 μg/kg to 2 mg/kg, and 
       wherein administration of the pharmaceutical composition to the subject increases the level of regulatory T cells in the subject by at least 1.76-fold five days after administration relative to the level of regulatory T cells in the subject before treatment with the pharmaceutical composition. 
     
     
         16 . The method of  claim 15 , wherein the human IL-2 variant protein domain comprises the N88R substitution relative to the amino acid sequence of SEQ ID NO: 2. 
     
     
         17 . The method of  claim 15 , wherein the peptide linker domain comprises the amino acid sequence of SEQ ID NO: 6. 
     
     
         18 . The method of  claim 15 , wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 4 or SEQ ID NO: 5. 
     
     
         19 . A method for treating an autoimmune disease, the method comprising administering to a subject in need thereof at least two doses of a pharmaceutical composition comprising a therapeutically-effective amount of a fusion protein comprising:
 a. a human IL-2 variant protein domain comprising a substitution selected from the group consisting of T3A and C125S relative to the amino acid sequence of SEQ ID NO: 2, and a substitution selected from the group consisting of D20H, N88I, N88G, N88R, Q126L, and Q126F relative to the amino acid sequence of SEQ ID NO: 2;   b. a peptide linker domain; and   c. an IgG Fc protein domain,   
       wherein each domain has an amino-terminus (N-terminus) and a carboxy terminus (C-terminus); and wherein the fusion protein is configured so that the C-terminus of the human IL-2 variant protein domain is fused through a peptide bond to the N-terminus of the peptide linker domain, and the N-terminus of the IgG Fc protein domain is fused through a peptide bond to the C-terminus of the peptide linker domain, 
       wherein the composition is administered to the subject at a dosing frequency from once every week to once every month and at a dose of 20 μg/kg to 2 mg/kg, 
       wherein administration of the pharmaceutical composition to the subject increases the ratio of regulatory T cells to conventional T cells (Treg/Tconv) by at least 2.9-fold six days after administration relative to the Treg/Tconv ratio in the subject before treatment with the pharmaceutical composition, 
       and wherein the autoimmune disease is selected from the group consisting of Psoriasis, Ulcerative Colitis, Crohn's disease, Pemphigus Vulgaris, Type 1 Diabetes, Systemic Lupus Erythematosus, Graft-versus-Host Disease, Autoimmune Vasculitis, Multiple Sclerosis, Amytrophic Lateral Sclerosis, Alopecia Areata, Uveitis, Duchenne Muscular Dystrophy, Scleroderma, and Neuromyelitis Optica. 
     
     
         20 . The method of  claim 19 , wherein the dosing frequency ranges from once every week to once every 2 weeks. 
     
     
         21 . The method of  claim 19 , wherein the human IL-2 variant protein domain comprises the N88R substitution relative to the amino acid sequence of SEQ ID NO: 2. 
     
     
         22 . The method of  claim 19 , wherein the human IL-2 variant protein domain comprises the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 3. 
     
     
         23 . The method of  claim 19 , wherein the peptide linker domain comprises the amino acid sequence of SEQ ID NO: 6. 
     
     
         24 . The method of  claim 19 , wherein the IgG Fc protein domain comprises the amino acid sequence of SEQ ID NO: 7. 
     
     
         25 . The method of  claim 19 , wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 4 or SEQ ID NO: 5. 
     
     
         26 . The method of  claim 19 , wherein the pharmaceutical composition is administered to the subject by subcutaneous administration.