Stereospecific determination of an HIV aspartyl protease inhibitor, PNU-103017, in rat, dog and human plasma using a Pirkle-concept high-performance liquid chromatographic column.
A sensitive stereospecific high-performance liquid chromatographic assay for the quantitation of the enantiomers of 4-cyano-N-(3-(cyclopropyl-(5,6,7,8,9,10-hexahydro-4-hydroxy-2-oxo-2H- cycloocta(b)pyran-3-yl)methyl)phenyl)benzenesulfonamide (PNU-103017) (I), an HIV protease inhibitor, in plasma of rat, dog and human was developed. The procedure involved an acetonitrile-aided protein precipitation followed by solid-phase extraction (SPE) of I from plasma into ethanol. Stereospecific separation was accomplished on a Pirkle-concept chiral column (Regis S,S-Whelk-01, 250x4.6 mm I.D.) with a mobile phase of absolute ethanol-0.1% acetic acid in hexane (30:70, v/v). The eluate was monitored by UV absorbance (295 nm). Linear calibration curves were obtained in the range of 0.2 to 500 microM, with a lower limit of quantitation of 0.1-0.2 microM for both enantiomers in either rat, dog or human plasma. Intra- and inter-assay precision and assay accuracy were demonstrated to be acceptable for the stereoselective pharmacokinetic analysis of I in plasma.
Williams
M G
MG
Drug Metabolism Research, Pharmacia and Upjohn, Inc., Kalamazoo, MI 49001, USA.
Zhong
W Z
WZ
D000818:Animals; D019380:Anti-HIV Agents / Q000097:blood*; D016282:Aspartic Acid Endopeptidases / Q000037:antagonists & inhibitors*; D002851:Chromatography, High Pressure Liquid; D004285:Dogs; D017320:HIV Protease Inhibitors / Q000097:blood*; D006801:Humans; D011753:Pyrones / Q000097:blood*; D051381:Rats; D015203:Reproducibility of Results; D013237:Stereoisomerism; D013449:Sulfonamides / Q000097:blood*
D019380:Anti-HIV Agents; D017320:HIV Protease Inhibitors; D011753:Pyrones; D013449:Sulfonamides; C107418:PNU 103017; D016282:Aspartic Acid Endopeptidases