title: Insulin analogues with modifications at position B26. Divergence of binding affinity and biological activity.
abstract: In this study, we prepared several shortened and full-length insulin analogues with substitutions at position B26. We compared the binding affinities of the analogues for rat adipose membranes with their ability to lower the plasma glucose level in nondiabetic Wistar rats in vivo after subcutaneous administration, and also with their ability to stimulate lipogenesis in vitro. We found that [NMeHisB26]-DTI-NH 2 and [NMeAlaB26]-DTI-NH 2 were very potent insulin analogues with respect to their binding affinities (214 and 465%, respectively, compared to that of human insulin), but they were significantly less potent than human insulin in vivo. Their full-length counterparts, [NMeHisB26]-insulin and [NMeAlaB26]-insulin, were less effective than human insulin with respect to binding affinity (10 and 21%, respectively) and in vivo activity, while [HisB26]-insulin exhibited properties similar to those of human insulin in all of the tests we carried out. The ability of selected analogues to stimulate lipogenesis in adipocytes was correlated with their biological potency in vivo. Taken together, our data suggest that the B26 residue and residues B26-B30 have ambiguous roles in binding affinity and in vivo activity. We hypothesize that our shortened analogues, [NMeHisB26]-DTI-NH 2 and [NMeAlaB26]-DTI-NH 2, have different modes of interaction with the insulin receptor compared with natural insulin and that these different modes of interaction result in a less effective metabolic response of the insulin receptor, despite the high binding potency of these analogues.
authors.affiliation: Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Flemingovo nám. 2, 166 10 Praha 6, Czech Republic.
authors.forename: Lenka
authors.initials: L
authors.lastname: Záková
authors.forename: Ludmila
authors.initials: L
authors.lastname: Kazdová
authors.forename: Ivona
authors.initials: I
authors.lastname: Hanclová
authors.forename: Eva
authors.initials: E
authors.lastname: Protivínská
authors.forename: Miloslav
authors.initials: M
authors.lastname: Sanda
authors.forename: Milos
authors.initials: M
authors.lastname: Budesínský
authors.forename: Jirí
authors.initials: J
authors.lastname: Jirácek
mesh_terms: D017667:Adipocytes / Q000378:metabolism; D000273:Adipose Tissue / Q000378:metabolism; D000818:Animals; D006801:Humans; D007328:Insulin / Q000737:chemistry* / Q000378:metabolism; D050155:Lipogenesis; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D008954:Models, Biological; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D051381:Rats; D017208:Rats, Wistar; D011972:Receptor, Insulin / Q000378:metabolism; D013552:Swine
chemical_list: D007328:Insulin; D011972:Receptor, Insulin