title: Design of a selective insulin receptor tyrosine kinase inhibitor and its effect on glucose uptake and metabolism in intact cells.
abstract: An inhibitor of the insulin receptor tyrosine kinase (IRTK), (hydroxy-2-naphthalenyl-methyl) phosphonic acid, was designed and synthesized and was shown to be an inhibitor of the biological effects of insulin in vitro. With a wheat germ purified human placental insulin receptor preparation, this compound inhibited the insulin-stimulated autophosphorylation of the 95-kDa beta-subunit of the insulin receptor (IC50 = 200 microM). The ability of the kinase to phosphorylate an exogenous peptide substrate, angiotensin II, was also inhibited. Half-maximal inhibition of basal and insulin-stimulated human placental IRTK activity was found at concentrations of 150 and 100 microM, respectively, with 2 mM angiotensin II as the peptide substrate. The inhibitor was found to be specific for tyrosine kinases over serine kinases and noncompetitive with ATP. The inhibitor was converted into various (acyloxy)methyl prodrugs in order to achieve permeability through cell membranes. These prodrugs inhibited insulin-stimulated autophosphorylation of the insulin receptor 95-kDa beta-subunit in intact CHO cells transfected with human insulin receptor. Inhibition of insulin-stimulated glucose oxidation in isolated rat adipocytes and 2-deoxyglucose uptake into CHO cells was observed with these prodrugs. Our data provide additional evidence for the involvement of the insulin receptor tyrosine kinase in the regulation of glucose uptake and metabolism. These results and additional data reported herein suggest that this class of prodrugs and inhibitors will be useful for modulating the activity of a variety of tyrosine kinases.
authors.affiliation: Department of Biochemical Endocrinology, Merck Sharp & Dohme Research Laboratories, Rahway, New Jersey 07065.
authors.forename: R
authors.initials: R
authors.lastname: Saperstein
authors.forename: P P
authors.initials: PP
authors.lastname: Vicario
authors.forename: H V
authors.initials: HV
authors.lastname: Strout
authors.forename: E
authors.initials: E
authors.lastname: Brady
authors.forename: E E
authors.initials: EE
authors.lastname: Slater
authors.forename: W J
authors.initials: WJ
authors.lastname: Greenlee
authors.forename: D L
authors.initials: DL
authors.lastname: Ondeyka
authors.forename: A A
authors.initials: AA
authors.lastname: Patchett
authors.forename: D G
authors.initials: DG
authors.lastname: Hangauer
mesh_terms: D000273:Adipose Tissue / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001693:Biological Transport, Active / Q000187:drug effects; D002460:Cell Line; D003837:Deoxy Sugars / Q000378:metabolism*; D003847:Deoxyglucose / Q000378:metabolism*; D015195:Drug Design; D005260:Female; D006801:Humans; D007328:Insulin / Q000494:pharmacology; D007700:Kinetics; D046911:Macromolecular Substances; D008297:Male; D008969:Molecular Sequence Data; D009281:Naphthalenes / Q000138:chemical synthesis / Q000494:pharmacology*; D063065:Organophosphonates; D009943:Organophosphorus Compounds / Q000138:chemical synthesis / Q000494:pharmacology*; D010766:Phosphorylation; D010920:Placenta / Q000378:metabolism; D011247:Pregnancy; D011355:Prodrugs / Q000494:pharmacology; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors*; D011817:Rabbits; D051381:Rats; D011919:Rats, Inbred Strains; D011972:Receptor, Insulin / Q000378:metabolism
chemical_list: D003837:Deoxy Sugars; D007328:Insulin; D046911:Macromolecular Substances; D009281:Naphthalenes; D063065:Organophosphonates; D009943:Organophosphorus Compounds; D011355:Prodrugs; C060935:(hydroxy-2-naphthalenylmethyl)phosphonic acid; D003847:Deoxyglucose; D011505:Protein-Tyrosine Kinases; D011972:Receptor, Insulin