THE ANALYTICAL CHEMISTRY OF ANTI-AIDS AGENTS

该项目的目标是研究和开发 合适的生物分析方法：（1）建立结构， 潜在的抗艾滋病药物和新的抗病毒药物的纯度，（2） 确定物理、化学和生物化学性质，包括 正辛醇-水分配系数，这些化合物及其 代谢物，和（3）测量这些药物及其代谢物， 生物样品，以阐明药理学和确定 药代动力学 高效液相色谱法（HPLC）和 质谱是重点技术。 I期药物 2 '-β-氟-2'，3 '-双脱氧腺苷（F-ddA）及其脱氨基化合物 抗HIV活性代谢物2 '-β-氟-2'，3 '-双脱氧肌苷（F-ddI） 仍然是主要关注的化合物。 分析战略 已开发并验证了采用反相HPLC的 F-ddA的常规和超灵敏测量， 感染艾滋病毒的人体体液。荧光衍生化和 使用荧光检测的HPLC分析允许测量F-ddA 在血浆中的浓度为纳摩尔。 代谢、分布和 F-ddA的药代动力学正在研究中， 在该药剂的I期临床试验期间口服给药 在艾滋病患者中。 初步结果显示， 口服F-ddA的生物利用度良好（>50%）。 亲脂性前药 腺苷脱氨酶激活的F-ddI也继续在 调查 用于测量的HPLC方法 生物体液和组织中的6-氯-2 '，3'-二脱氧嘌呤核苷 已经被开发并应用于显示中枢神经系统的增强 给药后大鼠中F-ddI的系统渗透 前药 细胞提取物的直接荧光衍生化 结合配对离子HPLC已被用于 亚皮摩尔量的细胞内 F-ddATP，F-ddA和F-ddI的活性代谢产物。

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 the 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 (HPLC) and mass spectrometry are the emphasized techniques. The Phase I drug 2'-beta-fluoro-2',3'-dideoxyadenosine (F-ddA) and its deaminated anti-HIV-active metabolite 2'-beta-fluoro-2',3'-dideoxyinosine (F-ddI) remain the compounds of primary interest. Analytical strategies employing reversed-phase HPLC have been developed and validated for both the routine and ultrasensitive measurement of F-ddA in HIV-infected human biological fluids. Fluorogenic derivatization and HPLC analysis with fluorescence detection allow measurement of F-ddA at nanomolar levels in plasma. The metabolism, distribution and pharmacokinetics of F-ddA is under investigation following intravenous and oral administration during a Phase I clinical trial of this agent in AIDS patients. Preliminary results indicate that the bioavailability of oral F-ddA is good (>50%). Lipophilic prodrugs of F-ddI activated by adenosine deaminase also continue under investigation. HPLC methodology for the measurement of 6-chloro-2',3'-dideoxypurineriboside in biological fluids and tissues has been developed and applied to show enhancement of central nervous system penetration of F-ddI in rats after administration of this prodrug. 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, the active metabolite of both F-ddA and F-ddI.