The objective of this project is the research and development of suitable bioanalytical methods to: (1) establish the structure and purity of potential anti-AIDS agents and new antiviral drugs, (2) determine physical, chemical and biochemical properties, including octanol-water partition coefficients, of these compounds and their metabolites, and (3) measure these drugs and their metabolites in biological samples to elucidate pharmacology and to determine pharmacokinetics. High- performance liquid chromatography and mass spectrometry are emphasized techniques. Sugar-fluorinated dideoxynucleosides, especially the Phase I clinical candidate 2'-F-ddA and its 2'-F-ddI metabolite, are currently the compounds of primary interest. Lipophilic prodrugs of F-ddI activated by adenosine deaminase also continue under investigation. HPLC assays for F-ddA involving both UV and fluorescence detection have been modified and validated for the analysis of HIV-deactivated human samples. Direct fluorogenic derivatization of cellular extracts in conjunction with paired-ion HPLC has been employed for the nonradiochemical measurement of subpicomole amounts of intracellular F-ddATP.
