Chimeric molecules comprising il-12 agonist polypeptide

A chimeric molecule, which comprises an IL-12 p40 subunit (p40), an IL-12 p35 subunit (p35), a masking moiety (MM), and a carrier (C);

• wherein:
• (a) said IL-12 subunit p40 comprises an amino acid sequence of SEQ ID NO:
• 5, or an amino acid sequence with at least 95% identity to that of SEQ ID NO:
• 5;
• (b) said IL-12 subunit p35 comprises an amino acid sequence of SEQ ID NO:
• 6, or an amino acid sequence with at least 95% identity to that of SEQ ID NO:
• 6;
• (c) said masking moiety comprises an amino acid sequence of SEQ ID NO:
• 1, 2, 3 or 4, or an amino acid sequence with at least 95% identity to that of SEQ ID NO:
• 1, 2, 3 or 4;
• (d) said carrier is selected from an albumin, an albumin fragment, an Fc domain, and an antibody;
• and
• (e) said IL-12 p40 subunit comprises mutations Lys260Ala and Arg261Ala.

A chimeric molecule, which comprises an IL-12 p40 subunit (p40), an IL-12 p35 subunit (p35), a masking moiety (MM), and a carrier (C);

• wherein:
• (a) said IL-12 subunit p40 comprises an amino acid sequence of SEQ ID NO:
• 46, 63, or 98, or an amino acid sequence with at least 99% identity to that of SEQ ID NO:
• 46, 63, or 98;
• (b) said IL-12 subunit p35 comprises an amino acid sequence of SEQ ID NO:
• 6 or an amino acid sequence with at least 95% identity to that of SEQ ID NO:
• 6;
• (c) said masking moiety comprises an amino acid sequence of SEQ ID NO:
• 1, 2, 3 or 4 or an amino acid sequence with at least 95% identity to that of SEQ ID NO:
• 1, 2, 3 or 4;
• and
• (d) said carrier is selected from an albumin or albumin fragment, an Fc domain, and an antibody.

A chimeric molecule, which comprises an IL-12 p40 subunit (p40), an IL-12 p35 subunit (p35), a masking moiety (MM), and a carrier (C);

• wherein:
• (a) said IL-12 subunit p40 comprising an amino acid sequence of SEQ ID NO:
• 5 or an amino acid sequence of at least 95% identity to that of SEQ ID NO:
• 5;
• (b) said IL-12 subunit p35 comprising an amino acid sequence of SEQ ID NO:
• 6 or an amino acid sequence of at least 95% identity to that of SEQ ID NO:
• 6;
• (c) said masking moiety comprises an amino acid sequence of SEQ ID NOS:
• 1, 2, 3 or 4, or an amino acid sequence of at least 95% identity to that of SEQ ID NO:
• 1, 2, 3 or 4;
• (d) said carrier comprises an antigen-binding moiety which binds to PD-1 and an Fc domain with a first Fc polypeptide and a second Fc polypeptide;
• (e) said masking moiety if fused to the C-terminus of the first Fc polypeptide with or without a non-cleavable or cleavable peptide linker;
• wherein said peptide linker optionally comprises an amino acid sequence of 3 amino acids to 22 amino acids in length;
• (f) said p40 subunit is fused to the 2nd Fc polypeptide optionally through a non-cleavable peptide linker;
• (g) said p35 subunit is fused to the C-terminus of the p40 subunit optionally through a peptide linker
• wherein said chimeric molecule has higher activity stimulating a PD-1 positive (or PD-1+) T cell compared to that of stimulating an NK cell.