Quantification of a dual angiotensin I-converting enzyme-neutral endopeptidase inhibitor and the active thiol metabolite in dog plasma by high-performance liquid chromatography with ultraviolet absorbance detection.
MDL 100,240 ([4S-[4 alpha,7 alpha(R*), 12b beta]]-7-[[2- (acetylthio)-1-oxo-3-phenylpropyl]amino]-1,2,3,4,6,7,8,12b-octahyd ro-6-oxo- pyrido[2,1-a][2]benzazepine-4-carboxylic acid, I) is the thioacetyl prodrug of the active thiol, MDL 100,173 (II), a dual inhibitor of angiotensin-I converting enzyme (ACE) and neutral endopeptidase (NEP). A drug which simultaneously inhibits both ACE and NEP may provide a unique therapy for hypertension and congestive heart failure. Methods based on high-performance liquid chromatography with UV absorbance detection at 200 nm were developed to support preclinical pharmacokinetic investigations. One method is used to measure unchanged I and free II, while the second method is used to quantify the total level of the thiol II after the plasma is incubated with the disulfide reducing agent, dithiothreitol. By either method, the analytes are quantified over the range of 25-1000 ng/ml with good accuracy and precision. The overall extraction efficiencies of unchanged I and free II in dog plasma were 79% and 86%, respectively, while the extraction efficiency of total II averaged 75%. Described in this report are the results obtained in validating the assay methods for measuring the compounds in plasma. Pharmacokinetic data are presented which were obtained by applying these methods to plasma collected from dogs dosed with I.
D000806:Angiotensin-Converting Enzyme Inhibitors / Q000097:blood* / Q000493:pharmacokinetics; D000818:Animals; D001552:Benzazepines / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004285:Dogs; D004355:Drug Stability; D005615:Freezing; D015260:Neprilysin / Q000037:antagonists & inhibitors*; D009994:Osmolar Concentration; D007703:Peptidyl-Dipeptidase A / Q000378:metabolism*; D011480:Protease Inhibitors / Q000097:blood* / Q000493:pharmacokinetics; D011725:Pyridines / Q000097:blood*; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet; D013438:Sulfhydryl Compounds / Q000097:blood
D000806:Angiotensin-Converting Enzyme Inhibitors; D001552:Benzazepines; D011480:Protease Inhibitors; D011725:Pyridines; D013438:Sulfhydryl Compounds; C085683:MDL 100240; C085684:MDL 100173; D007703:Peptidyl-Dipeptidase A; D015260:Neprilysin