Studies of intramolecular rearrangements of acyl-linked glucuronides using salicylic acid, flufenamic acid, and (S)- and (R)-benoxaprofen and confirmation of isomerization in acyl-linked delta 9-11-carboxytetrahydrocannabinol glucuronide.
NMR and HPLC have been used to investigate the rearrangements of 1-O-acylglucuronides in vitro and the occurrence of rearranged isomers in urine. Glucuronides of flufenamic acid, (S)- and (R)-benoxaprofen, salicylic acid, and delta 9-11-carboxytetrahydrocannabinol were synthesized, by use of immobilized enzymes, or purified from urine. Ester-linked isomers of these gluruconides were characterized, and isomers derived from flufenamic acid, (S)-benoxaprofen, and salicylic acid were purified for further study by NMR and HPLC. The positions of the new ester linkages could be identified by two-dimensional NMR. Shifts not only in the resonance of the proton adjacent to the esterified hydroxyl group but also in the resonance of the anomeric proton on carbon 1 of the glucuronic acid moiety could be correlated with the position of each isomeric ester bond. HPLC elution times also correlated with ester position in this small set of samples. The sequences of isomer formation were studied in situ by NMR and also at pH 8 by HPLC. These studies indicate that, for the three cases examined, the C-2 ester is formed first, followed by formation of C-3 and C-4 esters. The purified isomeric esters were found not to re-form the high-energy 1-O-acyl bond. All other rearrangement steps are reversible. In contrast to other glycosides and glycerol esters, no evidence could be found for rearrangements beyond nearest-neighbor hydroxyl groups in glucuronic acid. The sequence of formation and reversibility is consistent with an ortho ester intermediate, as has been proposed for rearrangements of other glycosides.(ABSTRACT TRUNCATED AT 250 WORDS)
Bradow
G
G
Johns Hopkins University, Baltimore, Maryland 21205.
Kan
L S
LS
Fenselau
C
C
D000818:Animals; D002241:Carbohydrates / Q000652:urine; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D013759:Dronabinol / Q000031:analogs & derivatives* / Q000737:chemistry; D005439:Flufenamic Acid / Q000737:chemistry* / Q000652:urine; D005965:Glucuronates / Q000737:chemistry*; D006801:Humans; D007202:Indicators and Reagents; D007536:Isomerism; D008253:Macaca mulatta; D013058:Mass Spectrometry; D011422:Propionates / Q000737:chemistry* / Q000652:urine; D011817:Rabbits; D012459:Salicylates / Q000737:chemistry* / Q000652:urine; D020156:Salicylic Acid; D013056:Spectrophotometry, Ultraviolet
D002241:Carbohydrates; D005965:Glucuronates; D007202:Indicators and Reagents; D011422:Propionates; D012459:Salicylates; C011677:benoxaprofen; C016780:11-nor-delta(9)-tetrahydrocannabinol-9-carboxylic acid; D005439:Flufenamic Acid; D013759:Dronabinol; D020156:Salicylic Acid