Bioactivity of glycogen phosphorylase inhibitors that bind to the purine nucleoside site

• Published in: Bioorganic & medicinal chemistry 2006-12, Vol.14 (23), p.7835-7845
• Main Authors: Hampson, Laura J., Arden, Catherine, Agius, Loranne, Ganotidis, Minas, Kosmopoulou, Magda N., Tiraidis, Costas, Elemes, Yiannis, Sakarellos, Constantinos, Leonidas, Demetres D., Oikonomakos, Nikos G.
• Format: Article
• Language: English
• Subjects: Adenosine Monophosphate - analogs & derivatives; Adenosine Monophosphate - metabolism; Alkenes - chemical synthesis; Alkenes - pharmacology; Allosteric Regulation; Binding Sites; Biological and medical sciences; Enzyme Inhibitors - metabolism; Flavonoids - chemical synthesis; Flavonoids - pharmacology; Flavopiridols; General and cellular metabolism. Vitamins; Glycogen - biosynthesis; Glycogen metabolism; Glycogen phosphorylase; Glycogen Phosphorylase - antagonists & inhibitors; Glycogenolysis - drug effects; Hepatocytes; Hepatocytes - enzymology; Hepatocytes - metabolism; Humans; Inhibitors; Medical sciences; Pharmacology. Drug treatments; Piperidines - chemical synthesis; Piperidines - pharmacology; Purine Nucleosides - metabolism; Type-2 diabetes
• ISSN: 0968-0896; 1464-3391
• DOI: 10.1016/j.bmc.2006.07.060

Four olefin derivatives of flavopiridol were synthesized (1–4). All derivatives are potent inhibitors of glycogen phosphorylase-b. Compounds 1 and 4 inhibit glycogenolysis in hepatocytes by allosteric inhibition and by counteraction of the activation of phosphorylase-b by phosphorylase kinase. 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-β-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.