Conjugates of fusion proteins of GLP-1 and FGF21

A polypeptide conjugate comprising a fusion polypeptide conjugated to at least one clearance reducing moiety (CRM),

• wherein the fusion polypeptide comprises or consists essentially of, from N-terminus to C terminus, a GLP-1, a polypeptide linker and a FGF21;
• wherein the GLP-1 comprises no more than 4 substitutions relative to native human GLP-1(7-37) while retaining substantial biological activity of the native human GLP-1(7-37), and comprises one or more substitutions selected from the group consisting of:
• A8G, K34R, G22E, and R36G, or any combination thereof, wherein the sequence of the native human GLP-1(7-37) is set forth in SEQ ID NO:
• 50;
• wherein the FGF21 comprises a combination of mutations relative to SEQ ID NO:
• 36 selected from the group consisting of:
• 1) 121Q, 168L, 171C, 180E;
• 2) 121Q, 168L, 171G, 174C, 180E;
• 3) 121Q, 168L, 171G, 180C;
• 4) 121Q, 168L, 171G, 180E;
• 5) 71C, 121Q, 168L, 171G, 180E;
• and 6) 121Q, 168L, 180E, 171G, ins 182C;
• wherein the at least one CRM is conjugated to at least one conjugatable residue in the fusion polypeptide;
• and
• wherein the polypeptide linker has a length of at least 4-120 amino acid residues, and
• wherein the fusion polypeptide comprises up to two conjugatable residues,
• wherein the fusion polypeptide comprises a first conjugatable residue and a second conjugatable residue, wherein:
• the first conjugatable residue is present in the FGF21 and the second conjugatable residue is present in the polypeptide linker,
• wherein both the first and the second conjugatable residues are cysteine;
• and
• wherein the conjugatable residue in the polypeptide linker is the most C-terminal residue of the polypeptide linker, or is at least 5 residues away (inclusive of the conjugatable residue) from the most C terminal residue of the GLP-1.