THE ANALYTICAL CHEMISTRY OF ANTI-AIDS AGENTS

本项目的目标是研究和开发 适用的生物分析方法：(1)建立结构和 潜在抗艾滋病药物和抗病毒新药的纯度(2) 确定物理、化学和生化特性，包括 这些化合物的辛醇-水分配系数以及它们的 代谢物，以及(3)测量这些药物及其代谢产物 生物样品用于阐明药理学和确定 药物动力学。高效液相色谱(HPLC)和 质谱仪是重点研究的技术。第一阶段的药物 2‘-β-氟-2’，3‘-二脱氧腺苷及其脱氨基 抗HIV活性代谢产物2‘-β-氟-2’，3‘-二脱氧肌苷(F-DDI) 仍然是主要感兴趣的化合物。分析策略 已经开发并验证了使用反相高效液相的 F-Dda的常规和超灵敏测定 感染艾滋病毒的人体生物体液。荧光衍生化和 高效液相色谱荧光检测法测定F-Dda 在纳摩尔水平的血浆中。新陈代谢、分布和 F-Dda静脉注射后的药代动力学研究 在该药的I期临床试验期间口服给药 在艾滋病患者身上。初步结果显示， 口服F-Dda的生物利用度良好(50%)。亲脂性前体药物 由腺苷脱氨酶激活的F-ddi也在 调查。高效毛细管电泳法测定血药浓度 生物液和组织中的6-氯-2‘，3’-二脱氧尿苷 已被开发并应用于显示中枢神经系统的增强 F-DDI给药后大鼠的系统渗透性 前药。细胞提取物的直接荧光衍生化 与成对离子高效液相色谱联用进行了分析 细胞内亚皮卡摩尔量的非放射化学测量 F-ddATP是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.