title: Structure and pharmaceutical formulation development of a new long-acting recombinant human insulin analog studied by NMR and MS.
abstract: A monomer structure of a novel human insulin analog A22S-B3K-B31R(SK3R) has been characterized by NMR in water/acetonitrile solution and compared with the structure of human insulin (HIS) established in the same medium. The composition of the oligomer ensemble for neat insulins in water was qualitatively assessed by monitoring, derived from NMR experiment, translational diffusion coefficient Dix10-10m2s-1, whose value is a population averaged of individual coefficients for species in oligomeric ensemble. Nanospray ESI/MS experiment was used to establish the masses of oligomers in pharmaceutical formulation of the SK3R insulin. The pharmacodynamic data were established and compared to insulin glargine characterized by the same profile of action in diabetics. The oligomerization process of insulin during development of pharmaceutical formulation with routinely used excipients has been studied using translation diffusion coefficient Dix10-10m2s-1established in water solution. These properties were compared with those of human insulin (HIS) which is a standard reference for novel recombinant insulins.
authors.affiliation: National Medicines Institute, 00-725 Warsaw, Poland.
authors.forename: Elżbieta
authors.initials: E
authors.lastname: Bednarek
authors.affiliation: National Medicines Institute, 00-725 Warsaw, Poland.
authors.forename: Jerzy
authors.initials: J
authors.lastname: Sitkowski
authors.affiliation: National Medicines Institute, 00-725 Warsaw, Poland.
authors.forename: Wojciech
authors.initials: W
authors.lastname: Bocian
authors.affiliation: Institute of Biotechnology and Antibiotics, Warsaw, Poland.
authors.forename: Piotr
authors.initials: P
authors.lastname: Borowicz
authors.affiliation: Institute of Biotechnology and Antibiotics, Warsaw, Poland.
authors.forename: Grażyna
authors.initials: G
authors.lastname: Płucienniczak
authors.affiliation: Institute of Biotechnology and Antibiotics, Warsaw, Poland.
authors.forename: Dorota
authors.initials: D
authors.lastname: Stadnik
authors.affiliation: Institute of Biotechnology and Antibiotics, Warsaw, Poland.
authors.forename: Weronika
authors.initials: W
authors.lastname: Surmacz-Chwedoruk
authors.affiliation: National Medicines Institute, 00-725 Warsaw, Poland.
authors.forename: Beata
authors.initials: B
authors.lastname: Jaworska
authors.affiliation: National Medicines Institute, 00-725 Warsaw, Poland. Electronic address: l.kozerski@nil.gov.pl.
authors.forename: Lech
authors.initials: L
authors.lastname: Kozerski
mesh_terms: D018360:Crystallography, X-Ray / Q000379:methods; D003692:Delayed-Action Preparations / Q000032:analysis / Q000737:chemistry; D004339:Drug Compounding; D006801:Humans; D007328:Insulin / Q000032:analysis* / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D000072756:Protein Conformation, alpha-Helical; D000072757:Protein Conformation, beta-Strand; D011994:Recombinant Proteins / Q000032:analysis / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods
chemical_list: D003692:Delayed-Action Preparations; D007328:Insulin; D011994:Recombinant Proteins
keywords: Nanospray ESI–MS; PFGSE NMR; Pharmaceutical form; Recombinant insulin