Single-chain insulin agonists exhibiting high activity at the insulin receptor

摘要

Single chain insulin analogs are provided having high potency and specificity for the insulin receptor. As disclosed herein optimally sized linking moieties can be used to link human insulin A and B chains, or analogs or derivatives thereof, wherein the carboxy terminus of the B25 amino acid of the B chain is linked to the amino terminus of the A1 amino acid of the A chain via the intervening linking moiety. In on embodiment the linking moiety comprises a polyethylene glycol of 6-16 monomer units and in an alternative embodiment the linking moiety comprises a non-native amino acid sequence derived form the IGF-1 C-peptide and comprising at least 8 amino acids and no more than 12 amino acid in length. Also disclosed are prodrug and conjugate derivatives of the single chain insulin analogs.

主权项

1. A single chain insulin agonist analog comprising the general structure B-LM-A wherein
B represents an insulin B chain comprising the sequence R22-X25LCGX29X30LVX33X34LX36LVCGX41X42GFX45 (SEQ ID NO: 20); A represents an insulin A chain comprising the sequence GIVX4X5CCX8X9X10CX12LX14X15LX17X18X19CX21—R13 (SEQ ID NO: 22); and LM represents a linking moiety linking the carboxy terminus of the B chain to the amino terminus of the A chain; further wherein the linking moiety is an 8 amino acid sequence consisting of the sequence X51X52GSSSX57X58 (SEQ ID NO: 29), wherein X51 is selected from the group consisting of glycine, alanine, valine, leucine, isoleucine and proline; X52 is any amino acid other than tyrosine; X57 and X58 are independently selected from the group consisting of arginine, lysine and ornithine; X4 is glutamic acid or aspartic acid; X5 is glutamine or glutamic acid; X8 is histidine, threonine or phenylalanine; X9 is serine, arginine, lysine, ornithine or alanine; X10 is isoleucine or serine; X12 is serine or aspartic acid; X14 is tyrosine, arginine, lysine, ornithine or alanine; X15 is glutamine, glutamic acid, arginine, alanine, lysine, ornithine or leucine; X17 is glutamine, glutamic acid, arginine, aspartic acid or lysine, ornithine; X18 is methionine, asparagine, glutamine, aspartic acid, glutamic acid or threonine; X19 is tyrosine, 4-methoxy-phenylalanine or 4-amino phenylalanine; X21 is selected from the group consisting of alanine, glycine, serine, valine, threonine, isoleucine, leucine, glutamine, glutamic acid, asparagine, aspartic acid, histidine, tryptophan, tyrosine, and methionine; X25 is selected from the group consisting of histidine and threonine; X29 is selected from the group consisting of alanine, glycine and serine; X30 is selected from the group consisting of histidine, aspartic acid, glutamic acid, homocysteic acid and cysteic acid; X33 is selected from the group consisting of aspartic acid and glutamic acid; X34 is selected from the group consisting of alanine and threonine; X36 is tyrosine; X41 is selected from the group consisting of glutamic acid, aspartic acid or asparagine; X42 is selected from the group consisting of alanine, ornithine, lysine and arginine; X45 is selected from the group consisting of tyrosine, histidine, asparagine and phenylalanine; wherein R13 is COOH or CONH2; and R22 is selected from the group consisting of AYRPSE (SEQ ID NO: 14), FVNQ (SEQ ID NO: 12), PGPE (SEQ ID NO: 11), a tripeptide glycine-proline-glutamic acid, a tripeptide valine-asparagine-glutamine, a dipeptide proline-glutamic acid, a dipeptide asparagine-glutamine, glutamine, glutamic acid and an N-terminal amine.