Protein and protein conjugate for diabetes treatment, and applications thereof

A fusion protein obtained by linking an interleukin-1 receptor antagonist protein or an analogue thereof with another polypeptide which is an insulin receptor binding polypeptide or an analogue thereof;

• the structure of said fusion protein is as follows:
• the interleukin-1 receptor antagonist protein or an analogue thereof —Lj— another polypeptide, or another polypeptide —Lj— the interleukin-1 receptor antagonist protein or an analogue thereof;
• the sequence of said interleukin-1 receptor antagonist protein or an analogue thereof is
• (SEQ ID NO:
• 204)
• XIL0RPSGRKSSKMQAFRIWDVNQKTFYLRNNQLVAGYLQGPNVNLEEKI
• DVVPIEPHALFLGIHGGKMXIL66LSXIL69VKSGDETRLQLEAV
• XIL84ITDLSENRKQDKRFAFIRSDSGPTTSFESAAXIL116PGWFL
• XIL122TAMEADQPVSLTNMPDEGVMVTKFYFQEDE,
• wherein
• XIL0 is absent, a cysteine, general formula 1, general formula 2, or methionine;
• XIL66, XIL69, XIL116, XIL122 is a cysteine or serine;
• XIL84 is a cysteine or asparagine;
• Lj is a linking group or spacer group, including a long chain fattyacid, polyethylene glycol, an amino acid, a short peptide, a protein, or a long chain formed by linking one or more of the optional longchain fatty acids, polyethylene glycol, amino acids, and short peptides through a covalent bond, or any of the structures connecting the two proteins/polypeptides through a covalent bond, or absent;
• the sequence of said insulin receptor binding polypeptide or an analogue thereof is
• (SEQ ID NO:
• 383)
• XIN107HLC[1]GSXIN108LVEALYLVC[2]GEXIN109GFXIN110
• XIN111XIN112XIN113XIN114XIN115-CL-GIVEQC[3]C[4]
• XIN127SIC[5]SLYQLENYC[6]XIN128XIN129 ,
• wherein
• XIN107 is a tetrapeptide of phenylalanine-valine-asparagine-glutamine, a tripeptide of valine-asparagine-glutamine, a dipeptide of asparagine-glutamine, glutamine, a sequence wherein any one of the amino acid residues in the sequence of the tetrapeptide, tripeptide, and dipeptide is substituted with a lysine or arginine, or absent;
• XIN108 is a histidine, phenylalanine, arginine, or glutamine;
• XIN109 is an arginine, alanine, glutamic acid, or aspartic acid;
• XIN110 is a phenylalanine, tyrosine, or histidine;
• XIN111 is a tyrosine, phenylalanine, or absent;
• XIN112 is a threonine, asparagine, or absent;
• XIN113 is a proline, lysine, glutamic acid, aspartic acid, or absent;
• XIN114 is a lysine, proline, arginine, glutamic acid, aspartic acid or absent;
• XIN115 is a threonine, or absent;
• XIN127 is a threonine, histidine, or arginine;
• XIN128 is an alanine, glycine, or asparagine;
• XIN129 is a lysine, a dipeptide of arginine-lysine, or absent;
• CL is a peptide sequence of 6-60 amino acids, wherein the amino acids are primarily selected from the group consisting of glycine, alanine, serine, threonine, and proline;
• the structure of general formula 1 is
• in which J is the structure of Ln-ML or a hydrogen atom;
• ML is a modifying group selected from —W—X—Y—Z, a fatty acid, polyethylene glycol, albumin, IgG Fc, glycosyl;
• Ln is a selectable linking group, covalent linkage or absent;
• the selectable linking group is selected from polyethylene glycol, a long chain fattyacid, a polypeptide, a natural or non-natural amino acid, or a longchain formed by linking one or more of polyethylene glycol molecules, fatty acids, amino acid molecules through covalent bonds;
• the structure of —W—X—Y—Z
• wherein
• W is an α-amino acid residue having a carboxyl group in the side chain, this residue forms, with one of its carboxyl groups, an amide group together with the amino group at the end of Ln;
• or W is a chain composed of two, three or four α-amino acids linked together via amide bonds, this chain is linked via an amide bond to the amino group at the end of Ln, the amino acid residues of W being selected from the group of amino acid residues having a neutral side chain and/or amino acid residues having a carboxyl group in the side chain so that W has at least one amino acid residue which has a carboxyl group in the side chain;
• or
• or W is a covalent bond from X to the amino group at the end of Ln;
• X is —CO—, —CH(COOH)CO—, —N(CH2COOH)CH2CO—, —N(CH2COOH)CH2CON—(CH2COOH)CH2CO—, —N(CH2CH2COOH)CH2CH2CO—, —NHCH(COOH)(CH2)4NHCO—, —N(CH2CH2COOH)CH2CH2CON(CH2CH2COOH)CH2CH2CO—, —N(CH2CH2COOH)CH2CO— or —N—(CH2COOH)CH2CH2CO—, wherein
• a) the above-mentioned X forms an amide bond with an amino group in W through the bond of the underlined carbonyl C, when W is an amino acid residue or a series of amino acid residues, or
• b) the above-mentioned X forms an amide bond with the amino group at the end of Ln through the bond of the underlined carbonyl C, when W is a covalent bond;
• Y is —(CH2)m, wherein m is an integer from 6 to 32;
• and
• Z is —COOH, —CO-Asp, —CO-Glu, —CO-Gly, —CO-Sar, —CH(COOH)2, —N(CH2COOH)2, —SO3H, —PO3H or absent, provided that Z is not —COOH when W is a covalent bond and X is —CO—;
• the structure of general formula 2 is
• wherein OL is Mr-Lr-NH-J, Mr-Lr-Z1, Mr-Lr-ML, or a hydrogen atom;
• J and ML are defined as the above;
• Mr is a functional group capable of reacting with the mercapto group to form a covalent linkage;
• Lr is an alternative linking group, covalent linkage, or absent, the alternative linking group is selected from polyethylene glycol, a long chain fattyacid, or a long chain formed by linking together one or more polyethylene glycol molecules and longchain fatty acid molecules through covalent bonds;
• Z1is —COOH, —CO-Asp, —CO-Glu, —CO-Gly, —CO-Sar, —CH(COOH)2, —N(CH2COOH)2, —SO3H, —PO3H or absent.