Identification and characterization of the major huperzine a metabolite in rat blood.
Huperzine A (Hup A) is under investigation as a treatment of Alzheimer's disease because of its properties of reversible and specific AChE inhibition. It has additional interesting pharmacological effects such as the protection of primary neuronal cells isolated from embryonic rat brains from glutamate-induced toxicity. We have isolated a new compound which has similar absorbance characteristics as Hup A from blood of rats administered Hup A. Monitoring the effluent from reversed-phase high-performance liquid chromatography (RP-HPLC) of blood collected 60 min after Hup A treatment at an absorbance of 308 nm (lambdamax for Hup A), yielded a peak height and area for this compound that was approximately 1.4-fold the initial Hup A peak. The compound was isolated from RP-HPLC fractions from blood and liver for analysis by mass spectrometry and nuclear magnetic resonance (NMR). The compound gave an (M+H)+ ion with m/z 259 in positive ion mode, yielding a molecular weight (MW) of 258. If derived from Hup A (MW 242), the change in MW indicates a mass gain of 16. This would be consistent with the addition of a single oxygen or a hydroxylation. To determine the location of the modification, it was examined by 1H NMR, and it was found that the added mass was due to a single epoxidation yielding 13,14-epoxy Hup-A.
Garcia
Gregory E
GE
Department of Biochemical Pharmacology, Division of Biochemistry, Walter Reed Army Institute of Research, Silver Spring, Maryland 20910-7500, USA. greg.garcia@na.amedd.army.mil
Hicks
Rickey P
RP
Skanchy
David
D
Moorad-Doctor
Deborah R
DR
Doctor
Bhupendra P
BP
Ved
Haresh S
HS
D000470:Alkaloids; D000818:Animals; D001274:Atmospheric Pressure; D002800:Cholinesterase Inhibitors / Q000008:administration & dosage / Q000097:blood / Q000493:pharmacokinetics*; D002851:Chromatography, High Pressure Liquid; D007273:Injections, Intramuscular; D018696:Neuroprotective Agents / Q000008:administration & dosage / Q000097:blood / Q000493:pharmacokinetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012717:Sesquiterpenes / Q000008:administration & dosage / Q000097:blood / Q000493:pharmacokinetics*; D021241:Spectrometry, Mass, Electrospray Ionization
D000470:Alkaloids; D002800:Cholinesterase Inhibitors; D018696:Neuroprotective Agents; D012717:Sesquiterpenes; C050426:huperzine A