ABSTRACT
A kappa opioid receptor binding inhibitor was isolated from the fermentation broth of a basidiomycete, Hericium ramosum CL24240 and identified as erinacine E (1). Three analogs of 1 were produced by fermentation in other media and by microbial biotransformation. Of these compounds, 1 was shown to be the most potent binding inhibitor. Preliminary SAR studies of these compounds indicated that all functional groups and side chains were required for the activity. Compound 1 was a highly-selective binding inhibitor for the kappa opioid receptor: 0.8 microM (IC50) for kappa, >200 microM for mu, and >200 microM for delta opioid receptor. Compound 1 suppressed electrically-stimulated twitch responses of rabbit vas deferens with an ED50 of 14 microM. The suppression was recovered by adding a selective kappa opioid receptor antagonist nor-binaltorphimine, indicating that 1 is a kappa opioid receptor agonist.
Subject(s)
Benzofurans/pharmacology , Receptors, Opioid, kappa/agonists , Spiro Compounds/pharmacology , Animals , Basidiomycota , Benzofurans/chemistry , Benzofurans/isolation & purification , Chromatography, High Pressure Liquid , Electric Stimulation , Fermentation , Guinea Pigs , Male , Rabbits , Receptors, Opioid, kappa/metabolism , Spiro Compounds/chemistry , Spiro Compounds/isolation & purification , Structure-Activity Relationship , Vas Deferens/drug effects , Vas Deferens/metabolismABSTRACT
The pathways involved in the metabolism of ketoaldonic acids by Erwinia sp. strain ATCC 39140 have been investigated by use of a combination of enzyme assays and isolation of bacterial mutants. The catabolism of 2,5-diketo-D-gluconate (2,5-DKG) to gluconate can proceed by two separate NAD(P)H-dependent pathways. The first pathway involves the direct reduction of 2,5-DKG to 5-keto-D-gluconate, which is then reduced to gluconate. The second pathway involves the consecutive reduction of 2,5-DKG to 2-keto-L-gulonate and L-idonic acid, which is then oxidized to 5-keto-D-gluconate, which is then reduced to gluconate. Gluconate, which can also be produced by the NAD(P)H-dependent reduction of 2-keto-D-gluconate, is phosphorylated to 6-phosphogluconate and further metabolized through the pentose phosphate pathway. No evidence was found for the existence of the Entner-Doudoroff pathway in this strain.