Novel conjugate of immune-stimulating il-2 analog and preparation method thereof

A long-acting conjugate of an interleukin-2 analog, represented by Chemical Formula 2 below:

• wherein X is an interleukin-2 analog comprising any one of polypeptide sequences selected from the amino acid sequence of SEQ ID NOS:
• 3 to 106;
• Z is a polyethylene glycol linker having a molecular weight of 2 kDa to 30 kDa;
• Fc is an immunoglobulin Fc region in the form of a dimer;
• and
• - indicates a covalent linkage between X and Z and between Z and Fc;
• wherein the long-acting conjugate is a long-acting conjugate in which one end of Z is covalently linked only to one polypeptide chain in the dimeric Fc region, and one molecule of X is covalently linked to the opposite end of Z.

A long-acting conjugate of an interleukin-2 analog, represented by Chemical Formula 3 below:

• wherein X′ is an interleukin-2 analog comprising an amino acid sequence represented by General Formula 1 below:
• [General Formula 1]
• (General Formula 1, SEQ ID NO:
• 212)
• X1-P-T-S-S-S-T-K-K-T-Q-L-Q-L-E-H-L-X18-X19-D-L-
• X22-M-I-L-N-G-I-N-N-Y-K-N-P-K-L-T-X38-M-L-T-X42-
• X43-F-X45-M-P-K-K-A-T-E-L-K-H-L-Q-C-L-E-X61-E-L-K-
• P-L-E-X68-V-L-N-L-A-X74-S-K-N-F-H-X80-X81-P-R-X84-
• X85-X86-S-N-I-N-X91-X92-V-X94-E-X96-K-G-S-E-T-T-F-
• M-C-E-Y-A-D-E-T-A-T-I-V-E-F-L-N-R-W-I-T-F-S-Q-S-I-
• I-S-T-L-T
• in General Formula 1 above,
• X1 is a deletion;
• X18 is leucine (L) or arginine (R);
• X19 is leucine (L) or tyrosine (Y);
• X22 is glutamic acid (E) or glutamine (Q);
• X38 is alanine (A), aspartic acid (D), or arginine (R);
• X42 is alanine (A), phenylalanine (F), lysine (K), or tryptophan (W);
• X43 is glutamic acid (E), lysine (K), or glutamine (Q);
• X45 is alanine (A) or tyrosine (Y);
• X61 is aspartic acid (D), glutamic acid (E), glutamine (Q), or arginine (R);
• X68 is aspartic acid (D) or glutamic acid (E);
• X74 is histidine (H) or glutamine (Q);
• X80 is phenylalanine (F), leucine (L), valine (V), or tyrosine (Y);
• X81 is aspartic acid (D), glutamic acid (E), or arginine (R);
• X84 is aspartic acid (D) or glutamic acid (E);
• X85 is alanine (A), glutamic acid (E), glycine (G), leucine (L), valine (V), tryptophan (W), or tyrosine (Y);
• X86 is alanine (A), glycine (G), isoleucine (I), or valine (V);
• X91 is threonine (T) or valine (V);
• X92 is phenylalanine (F), isoleucine (I), or tyrosine (Y);
• X94 is phenylalanine (F) or leucine (L);
• X96 is phenylalanine (F) or leucine (L);
• Z is a polyethylene glycol linker having a molecular weight of 2 kDa to 30 kDa;
• Fc is an immunoglobulin Fc region in the form of a dimer;
• and
• - indicates a covalent linkage between X′ and Z and between Z and Fc;
• wherein the long-acting conjugate is a long-acting conjugate in which one end of Z is covalently linked only to one polypeptide chain in the dimeric Fc region, and one molecule of X′ is covalently linked to the opposite end of Z.

A long-acting conjugate of an interleukin-2 analog, represented by Chemical Formula 4 below:

• wherein X″ is an interleukin-2 analog comprising an amino acid sequence represented by General Formula 2 below:
• [General Formula 2]
• (General Formula 2, SEQ ID NO:
• 213)
• X1-P-T-S-S-S-T-K-K-T-Q-L-Q-L-E-H-L-X18-L-D-L-X22-
• M-I-L-N-G-I-N-N-Y-K-N-P-K-L-T-X38-M-L-T-X42-K-F-Y-
• M-P-K-K-A-T-E-L-K-H-L-Q-C-L-E-X61-E-L-K-P-L-E-X68-
• V-L-N-L-A-Q-S-K-N-F-H-F-X81-P-R-D-X85-X86-S-N-I-N-
• V-F-V-L-E-L-K-G-S-E-T-T-F-M-C-E-Y-A-D-E-T-A-T-I-V-
• E-F-L-N-R-W-I-T-F-S-Q-S-I-I-S-T-L-T
• X1 is a deletion;
• X18 is leucine (L) or arginine (R);
• X22 is glutamic acid (E) or glutamine (Q);
• X38 is alanine (A) or arginine (R);
• X42 is phenylalanine (F) or lysine (K);
• X61 is aspartic acid (D) or glutamic acid (E);
• X68 is aspartic acid (D) or glutamic acid (E);
• X81 is aspartic acid (D) or glutamic acid (E);
• X85 is leucine (L) or valine (V);
• X86 is isoleucine (I) or valine (V);
• Z is a polyethylene glycol linker having a molecular weight of 2 kDa to 30 kDa;
• Fc is an immunoglobulin Fc region in the form of a dimer;
• and
• - indicates a covalent linkage between X″ and Z and between Z and Fc;
• wherein the long-acting conjugate is a long-acting conjugate in which one end of Z is covalently linked only to one polypeptide chain in the dimeric Fc region, and one molecule of X″ is covalently linked to the opposite end of Z.