title: Bioactivity of glycogen phosphorylase inhibitors that bind to the purine nucleoside site.
abstract: The bioactivity in hepatocytes of glycogen phosphorylase inhibitors that bind to the active site, the allosteric activator site and the indole carboxamide site has been described. However, the pharmacological potential of the purine nucleoside inhibitor site has remained unexplored. We report the chemical synthesis and bioactivity in hepatocytes of four new olefin derivatives of flavopiridol (1-4) that bind to the purine site. Flavopiridol and 1-4 counteracted the activation of phosphorylase in hepatocytes caused by AICAR (5-aminoimidazole-4-carboxamide 1-beta-D-ribofuranoside), which is metabolised to an AMP analogue. Unlike an indole carboxamide inhibitor, the analogues 1 and 4 suppressed the basal rate of glycogenolysis in hepatocytes by allosteric inhibition rather than by inactivation of phosphorylase, and accordingly caused negligible stimulation of glycogen synthesis. However, they counteracted the stimulation of glycogenolysis by dibutyryl cAMP by both allosteric inhibition and inactivation of phosphorylase. Cumulatively, the results show key differences between purine site and indole carboxamide site inhibitors in terms of (i) relative roles of dephosphorylation of phosphorylase-a as compared with allosteric inhibition, (ii) counteraction of the efficacy of the inhibitors on glycogenolysis by dibutyryl-cAMP and (iii) stimulation of glycogen synthesis.
authors.affiliation: School of Clinical Medical Sciences-Diabetes, Newcastle University, The Medical School, Newcastle upon Tyne NE2 4HH, UK.
authors.forename: Laura J
authors.initials: LJ
authors.lastname: Hampson
authors.forename: Catherine
authors.initials: C
authors.lastname: Arden
authors.forename: Loranne
authors.initials: L
authors.lastname: Agius
authors.forename: Minas
authors.initials: M
authors.lastname: Ganotidis
authors.forename: Magda N
authors.initials: MN
authors.lastname: Kosmopoulou
authors.forename: Costas
authors.initials: C
authors.lastname: Tiraidis
authors.forename: Yiannis
authors.initials: Y
authors.lastname: Elemes
authors.forename: Constantinos
authors.initials: C
authors.lastname: Sakarellos
authors.forename: Demetres D
authors.initials: DD
authors.lastname: Leonidas
authors.forename: Nikos G
authors.initials: NG
authors.lastname: Oikonomakos
mesh_terms: D000249:Adenosine Monophosphate / Q000031:analogs & derivatives / Q000378:metabolism; D000475:Alkenes / Q000138:chemical synthesis / Q000494:pharmacology; D000494:Allosteric Regulation; D001665:Binding Sites; D004791:Enzyme Inhibitors / Q000378:metabolism; D005419:Flavonoids / Q000138:chemical synthesis / Q000494:pharmacology; D006003:Glycogen / Q000096:biosynthesis; D024981:Glycogen Phosphorylase / Q000037:antagonists & inhibitors*; D050261:Glycogenolysis / Q000187:drug effects; D022781:Hepatocytes / Q000201:enzymology / Q000378:metabolism; D006801:Humans; D010880:Piperidines / Q000138:chemical synthesis / Q000494:pharmacology; D011684:Purine Nucleosides / Q000378:metabolism*
chemical_list: D000475:Alkenes; D004791:Enzyme Inhibitors; D005419:Flavonoids; D010880:Piperidines; D011684:Purine Nucleosides; D000249:Adenosine Monophosphate; C077990:alvocidib; D006003:Glycogen; D024981:Glycogen Phosphorylase