Heterodimeric proteins and uses thereof

Abstract

The present invention relates, inter alia, to compositions and methods, including heterodimeric proteins that find use in the treatment of disease, such as immunotherapies for cancer and autoimmunity.

1 .- 42 . (canceled) 
     
     
         43 . A heterodimeric protein, wherein the heterodimeric protein comprises a first polypeptide chain and a second polypeptide chain, wherein:
 the first polypeptide chain comprises a first subunit of interleukin (IL)-6 receptor linked by a first charge polarized core domain to a first subunit of IL-35; and   the second polypeptide chain comprises a second subunit of the IL-6 receptor linked by a second charge polarized core domain to a second subunit of the IL-35,   wherein:
 the first and second subunits of IL-35 are IL-12α and IL-27β; 
 the first and second subunits of IL-6 receptor are IL-6Rα and gp130; 
 the first charge polarized core domain and the second charge polarized core domain each independently comprise one of:
 a peptide comprising positively charged amino acid residues selected from histidine, lysine and arginine, or 
 a peptide comprising negatively charged amino acid residues selected from aspartic acid and glutamic acid, 
 
   wherein the first polypeptide chain and the second polypeptide chain associate through electrostatic interactions between the peptide comprising positively charged amino acid residues and the peptide comprising negatively charged amino acid residues to yield functional heterodimers of IL-35 and IL-6 receptor.   
     
     
         44 . The heterodimeric protein of  claim 43 , wherein the first and/or second charge polarized core domain comprises a synthetic linker comprising a polyethyleneglycol (PEG) molecule. 
     
     
         45 . The heterodimeric protein of  claim 43 , wherein the first and/or second charge polarized core domain comprises a polypeptide linker selected from a flexible amino acid sequence, IgG hinge region, or antibody sequence. 
     
     
         46 . The heterodimeric protein of  claim 45 , wherein the linker comprises a hinge-CH2-CH3 Fc domain derived from IgG1 or IgG4. 
     
     
         47 . The heterodimeric protein of  claim 43 , wherein the peptide comprising positively charged amino acid residues comprises the sequence RKGGKR (SEQ ID NO: 11) or GSGSRKGGKRGS (SEQ ID NO: 12). 
     
     
         48 . The heterodimeric protein of  claim 43 , wherein the peptide comprising negatively charged amino acid residues comprises the sequence DEGGED (SEQ ID NO: 13) or GSGSDEGGEDGS (SEQ ID NO: 14). 
     
     
         49 . The heterodimeric protein of  claim 43 , wherein the first and/or second charge polarized core domain comprises a hinge-CH2-CH3 Fc domain comprising one or more amino acid changes for promoting heterodimerization via increased hydrogen bonding and/or van der Waals forces. 
     
     
         50 . The heterodimeric protein of  claim 49 , wherein the one or more amino acid changes creates a knob in hole motif. 
     
     
         51 . The heterodimeric protein of  claim 50 , wherein
 the knob in hole motif is formed by one or more amino acid changes that replaces one or more tyrosine (Y) residues with one or more threonine (T) residues in the first charge polarized core domain comprises and/or is formed by one or more amino acid changes that replaces one or more threonine (T) residues with one or more tyrosine (Y) residues in the second charge polarized core domain, or   the knob in hole motif is formed by one or more amino acid changes that replaces one or more tyrosine (Y) residues with one or more threonine (T) residues in the second charge polarized core domain comprises and/or is formed by one or more amino acid changes that replaces one or more threonine (T) residues with one or more tyrosine (Y) residues in the first charge polarized core domain.   
     
     
         52 . The heterodimeric protein of  claim 49 , wherein one or both of the charge polarized core domains comprise one or more effector and complement silencing substitutions selected from L234A and L235A (LALA), and optionally, P329G, or one or both of the charge polarized core domains comprise one or more half-life extension mutations selected from M252Y, 5254T, and T256E. 
     
     
         53 . The heterodimeric protein of  claim 52 , wherein the first charge polarized core domain and the second charge polarized core domain comprises an amino acid sequence that is at least 95% identical to the amino acids sequence of SEQ ID NO: 24, and the other comprises an amino acid sequence that is at least 95% identical to the amino acids sequence of SEQ ID NO: 25. 
     
     
         54 . The heterodimeric protein of  claim 43 , wherein one of the first charge polarized core domain and the second charge polarized core domain comprises an amino acid sequence that is at least 95% identical to the amino acids sequence of SEQ ID NO: 16, and the other comprises an amino acid sequence that is at least 95% identical to the amino acids sequence of SEQ ID NO: 17. 
     
     
         55 . The heterodimeric protein of  claim 43 , wherein:
 the gp130 comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 18;   the IL-6Rα comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 19;   the IL-12α comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 20; and/or   the IL-27β comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 21.   
     
     
         56 . The heterodimeric protein of  claim 43 , wherein the first polypeptide chain and the second polypeptide chain comprise amino acid sequences that are at least 95% identical to the amino acid sequences of
 SEQ ID NO: 22 and SEQ ID NO: 23; or   SEQ ID NO: 34 and SEQ ID NO: 35.   
     
     
         57 . The heterodimeric protein of  claim 43 , wherein
 the first polypeptide chain comprises the first subunit of IL-6 receptor at the amino terminus linked by the first charge polarized core domain to the first subunit of IL-35 at the carboxy terminus; and   the second polypeptide chain comprises the second subunit of the IL-6 receptor at the amino terminus linked by the second charge polarized core domain to the second subunit of the IL-35 at the carboxy terminus.   
     
     
         58 . The heterodimeric protein of  claim 43 , wherein
 the first polypeptide chain comprises the first subunit of IL-35 at the amino terminus linked by the first charge polarized core domain to the first subunit of IL-6 receptor at the carboxy terminus; and   the second polypeptide chain comprises the second subunit of the IL-35 at the amino terminus linked by the second charge polarized core domain to the second subunit of the IL-6 receptor at the carboxy terminus.   
     
     
         59 . A nucleic acid encoding the first and/or second polypeptide chains of the heterodimeric protein of  claim 43 . 
     
     
         60 . An expression vector comprising the nucleic acid of  claim 59 .