A5085997583- Pancreatic function and diabetes
- Metabolism, Diabetes, and Cancer
- Diabetes Management and Research
- Growth Hormone and Insulin-like Growth Factors
- Glycosylation and Glycoproteins Research
- Monoclonal and Polyclonal Antibodies Research
- Fungal and yeast genetics research
- Diabetes Treatment and Management
- Chemical Synthesis and Analysis
- Protein purification and stability
- Transgenic Plants and Applications
- Cellular transport and secretion
- Protein Structure and Dynamics
- Receptor Mechanisms and Signaling
- Click Chemistry and Applications
- Diabetes and associated disorders
- Peptidase Inhibition and Analysis
- Endoplasmic Reticulum Stress and Disease
- Viral Infectious Diseases and Gene Expression in Insects
- Hyperglycemia and glycemic control in critically ill and hospitalized patients
- Computational Drug Discovery Methods
- Biofuel production and bioconversion
- Toxin Mechanisms and Immunotoxins
- Protein Interaction Studies and Fluorescence Analysis
- HER2/EGFR in Cancer Research
Novo Nordisk (Denmark)
2011-2024
Novo Nordisk (United Kingdom)
2009
Beppu University
1994
Weatherford College
1994
Bispebjerg Hospital
1993
Addenbrooke's Hospital
1992
University of Cambridge
1992
Max Planck Institute of Biochemistry
1992
Steno Diabetes Centers
1992
University of Washington
1988-1991
Introduction Insulin icodec is a novel, long-acting insulin analog designed to cover basal requirements with once-weekly subcutaneous administration. Here we describe the molecular engineering and biological pharmacological properties of icodec. Research design methods A number in vitro assays measuring receptor binding, intracellular signaling as well cellular metabolic mitogenic responses were used characterize To evaluate individuals type 2 diabetes, randomized, double-blind,...
Alanine scanning mutagenesis has been used to identify specific side chains of insulin which strongly influence binding the receptor. A total 21 new analog constructs were made, and in addition 7 high pressure liquid chromatography-purified analogs tested, covering alanine substitutions positions B1, B2, B3, B4, B8, B9, B10, B11, B12, B13, B16, B17, B18, B20, B21, B22, B26, A4, A8, A9, A12, A13, A14, A15, A16, A17, A19, A21. Binding data on revealed that mutations most disruptive for at...
Here, we describe the molecular engineering of insulin icodec to achieve a plasma half-life 196 h in humans, suitable for once-weekly subcutaneously administration. Insulin is based on re-engineering ultra-long oral basal OI338 with 70 humans. This systematic was accomplished by (1) further increasing albumin binding changing fatty diacid from 1,18-octadecanedioic acid (C18) 1,20-icosanedioic (C20) and (2) reducing receptor affinity B16Tyr → His substitution. selected screening long...
To identify the region(s) of insulin receptor and insulin-like growth factor I (IGF-I) responsible for ligand specificity (high-affinity binding), expression vectors encoding soluble chimeric insulin/IGF-I receptors were prepared. The expressed in mammalian cells partially purified. Binding studies revealed that a construct comprising an IGF-I which 68 N-terminal amino acids alpha-subunit had replaced equivalent segment displayed markedly increased affinity insulin. In contrast,...
Abstract Recently, the clinical proof of concept for first ultra-long oral insulin was reported, showing efficacy and safety similar to subcutaneously administered glargine. Here, we report molecular engineering as well biological pharmacological properties these analogues. Molecules were designed have pharmacokinetic profile minimize variability in plasma exposure. Elimination half-life ~20 h dogs ~70 man is achieved by a strong albumin binding, lowering receptor affinity 500-fold slow down...
Insulin icodec is a once-weekly insulin analogue that has long half-life of approximately 7 days, making it suitable for once weekly dosing. The molecule was developed based on the hypothesis lowering receptor affinity and introducing strong albumin-binding moiety would result in half-life, provided non-receptor-mediated clearance diminished. Here, we report an mechanism, resulting splitting molecules into its A-chain B-chain by thiol-disulphide exchange reaction. Even though substitutions...
We obtained 20 mouse monoclonal antibodies specific for human type I insulin-like growth factor (IGF) receptors, using transfected cells expressing high levels of receptors (IGF-1R/3T3 cells) as immunogen. The immunoprecipitated receptor.125I-IGF-I complexes and biosynthetically labeled from IGF-1R/3T3 but did not react with insulin or rat IGF receptors. Several stimulated DNA synthesis in cells, the maximum stimulation was only 25% that produced by IGF-I. fell into seven groups recognizing...
Insulin-like growth factor I (IGF-I) and insulin receptors are structurally similar with ligand-stimulated tyrosine kinase activity in their cytoplasmic domains. The function of the receptor signal transduction has been studied extensively contrast to IGF-I kinase. In present study we have analyzed regulatory carboxyl-terminal domains mitogenic signaling by overexpression mutant mouse NIH-3T3 fibroblasts. A receptor, which 3 tyrosines (Tyr1131, Tyr1135, Tyr1136) analogous three major...
Insulin is a key hormone controlling glucose homeostasis. All known vertebrate insulin analogs have classical structure with three 100% conserved disulfide bonds that are essential for structural stability and thus the function of insulin. It might be hypothesized an additional bond may enhance which would highly desirable in pharmaceutical use. To address this hypothesis, we designed interchain positions A10/B4 based on Cα-Cα distances, solvent exposure, side-chain orientation human (HI)...
Endogenous insulin secretion exposes the liver to three times higher concentrations than rest of body. Because subcutaneous delivery eliminates this gradient and is associated with metabolic abnormalities, functionally restoring physiologic may provide therapeutic benefits. The effects recombinant human (HI) delivered intraportally or peripherally were compared an acylated model compound (insulin-327) in dogs. During somatostatin basal portal vein glucagon infusion, was infused portally...
Peripheral hyperinsulinemia resulting from subcutaneous insulin injection is associated with metabolic defects that include abnormal glucose metabolism. The first aim of this study was to quantify the impairments in liver and muscle metabolism occur when delivered via a peripheral vein compared it given through its endogenous secretory route (the hepatic portal vein) overnight-fasted conscious dogs. second determine if delivery hepato-preferential analog could restore physiologic response...
The yeasts Pichia pastoris and Saccharomyces cerevisiae have similar overall features regarding the secretory expression of insulin. S. mating factor alpha (alpha-factor) prepro-leader facilitated secretion an insulin precursor, but not proinsulin expressed in P. pastoris. Synthetic prepro-leaders developed for precursor also In contrast with cerevisiae, only no unprocessed hyperglycosylated alpha-factor pro-leader/insulin fusion protein was secreted from A spacer peptide increased...
We have previously shown, using truncated soluble recombinant receptors, that substituting the 62 N-terminal amino acids of a subunit from insulinlike growth factor I receptor (IGFIR) with corresponding 68 insulin (IR) results in chimeric an approximately 200-fold increase affinity for and only 5- fold decrease insulin-like (IGFI)
ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTChanging the insulin receptor to possess insulin-like growth factor I ligand specificityAsser S. Andersen, Thomas Kjeldsen, Finn C. Wiberg, Per Michael Christensen, Jesper Skou Rasmussen, Kjeld Norris, Karin Bach Moeller, and Niels Peter H. MoellerCite this: Biochemistry 1990, 29, 32, 7363–7366Publication Date (Print):August 14, 1990Publication History Published online1 May 2002Published inissue 14 August...
An insulin-containing fusion protein (ICFP, encoding the yeast prepro-α factor leader peptide fused via a lysine-arginine cleavage site to single chain insulin) has been expressed in Saccharomyces cerevisiae where it is inefficiently secreted. Single gene disruptions have identified that cause enhanced immunoreactive insulin secretion (eis). Five out of six eis mutants prove be vacuolar sorting (vps)8, vps35, vps13, vps4, and vps36, which affect Golgi↔endosome trafficking. Indeed, wild-type...
Adaptation to efficient heterologous expression is a prerequisite for recombinant proteins fulfill their clinical and biotechnological potential. We describe rational strategy optimize the secretion efficiency in yeast of an insulin precursor by structure-based engineering folding stability. The yield fast-acting analogue (AspB28) expressed was enhanced 5-fold specific interaction between aromatic amino acid connecting peptide phenol binding site hydrophobic core molecule. This characterized...
Recently, the first basal oral insulin (OI338) was shown to provide similar treatment outcomes glargine in a phase 2a clinical trial. Here, we report engineering of novel class analogues which OI338, 10, this publication, successfully tested We found that introduction two substitutions, A14E and B25H, needed increased stability toward proteolysis. Ultralong pharmacokinetic profiles were obtained by attaching an albumin-binding side chain derived from octadecanedioic (C18) or icosanedioic...
The T-antigen system and the mean nuclear volume have been proposed as risk variables in bladder tumors. This study includes 34 patients with initially noninvasive (Ta) transitional cell carcinomas who experienced different courses of disease. Tissue specimens primary tumors were analyzed for expression T-antigen, Tn-antigen, sialosyl-Tn-antigen using monoclonal antibodies (MoAb) lectin peanut agglutinin (PNA) an indirect immunoperoxidase method. In addition, was estimated by morphometry....