Hydromorphone-3-glucuronide: biochemical synthesis and preliminary pharmacological evaluation.
Hydromorphone-3-glucuronide (H3G) was synthesized biochemically using rat liver microsomes, uridine-5'-diphosphoglucuronic acid (UDPGA) and the substrate, hydromorphone. Initially, the crude putative H3G product was purified by ethyl acetate precipitation and washing with acetonitrile. Final purification was achieved using semi-preparative high-performance-liquid-chromatography (HPLC) with ultraviolet (UV) detection. The purity of the final H3G product was shown by HPLC with electrochemical and ultraviolet detection to be > 99.9% and it was produced in a yield of = 60% (on a molar basis). The chemical structure of the putative H3G was confirmed by enzymatic hydrolysis of the glucuronide moiety using beta-glucuronidase, producing a hydrolysis product with the same HPLC retention time as the hydromorphone reference standard. Using HPLC with tandem mass spectrometry (HPLC-MS-MS) in the positive ionization mode, the molecular mass (M+1) was found to be 462 g/mol, in agreement with H3G's expected molecular weight of 461 g/mol. Importantly, proton-NMR indicated that the glucuronide moiety was attached at the 3-phenolic position of hydromorphone. A preliminary evaluation of H3G's intrinsic pharmacological effects revealed that following i.c.v. administration to adult male Sprague-Dawley rats in a dose of 5 microg, H3G evoked a range of excitatory behavioural effects including chewing, rearing, myoclonus, ataxia and tonic-clonic convulsions, in a manner similar to that reported previously for the glucuronide metabolites of morphine, morphine-3-glucuronide and normorphine-3-glucuronide.
School of Pharmacy, The University of Queensland, Brisbane, Australia.
A W
AW
Wright
M L
ML
Nocente
M T
MT
Smith
D017109:Akathisia, Drug-Induced / Q000209:etiology; D000701:Analgesics, Opioid / Q000378:metabolism; D000818:Animals; D001522:Behavior, Animal / Q000187:drug effects; D001921:Brain / Q000187:drug effects*; D002851:Chromatography, High Pressure Liquid; D004409:Dyskinesia, Drug-Induced / Q000209:etiology; D008401:Gas Chromatography-Mass Spectrometry; D005965:Glucuronates / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology*; D004091:Hydromorphone / Q000031:analogs & derivatives* / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D007276:Injections, Intraventricular; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D008862:Microsomes, Liver / Q000378:metabolism*; D009043:Motor Activity / Q000187:drug effects; D051381:Rats; D017207:Rats, Sprague-Dawley; D012640:Seizures / Q000139:chemically induced / Q000503:physiopathology; D014535:Uridine Diphosphate Glucuronic Acid / Q000378:metabolism
D000701:Analgesics, Opioid; D005965:Glucuronates; C093624:hydromorphone-3-glucuronide; D014535:Uridine Diphosphate Glucuronic Acid; D004091:Hydromorphone