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。高效液相色谱仪和质谱仪 光谱是重点研究的技术。第一阶段药物2‘-b- 氟-2‘，3’-二脱氧腺苷(F-Dda)及其脱氨基抗HIV活性 代谢产物2‘-b-氟-2’，3‘-二脱氧肌苷(F-DDI)仍然是化合物 主要利益所在。反相高效液相色谱法的分析策略 已开发、验证并应用于常规和 HIV感染者体内F-Dda的超灵敏测定 体液。荧光衍生化及其高效液相色谱分析 检测可以在纳摩尔水平上测量血浆中的F-Dda。这个 口服F-Dda的人体代谢、分布及药代动力学 是结合这一项目的I期临床试验确定的 成人艾滋病患者中的代理。口服液的口服生物利用度 无论是禁食(74%)还是进食，F-Dda的含量都很高 (%)。初步研究表明，F-Dda的生物利用度 以胶囊形式服用也很好(&gt；50%)。F-DDI的亲脂性前药 由腺苷脱氨酶激活的活性也在继续调查中。高效液相色谱仪 快速测定F-DDI程序的方法学 生物中的氯-2‘，3’-二脱氧尿苷及其代谢物 液体和组织正在开发中。直接荧光衍生化 细胞提取物与配对离子高效液相色谱联用 用于亚皮可摩尔量的非放射化学测量 胞内F-ddATP是F-dda和F-ddi的活性代谢产物。 F-ddATP水平将在外周血单核细胞中进行测量 来自接受F-Dda治疗的患者，并与观察到的抗HIV相关 活动。

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'-b- fluoro-2',3'-dideoxyadenosine (F-ddA) and its deaminated anti-HIV-active metabolite 2'-b-fluoro-2',3'-dideoxyinosine (F-ddI) remain the compounds of primary interest. Analytical strategies employing reversed-phase HPLC have been developed, validated and applied 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 human metabolism, distribution and pharmacokinetics of oral F-ddA is being determined in conjunction with a Phase I clinical trial of this agent in adult AIDS patients. The oral bioavailability of the liquid formulation of F-ddA is high after either fasting (74%)or with food (64%). Preliminary studies indicate that the bioavailability of F-ddA given as capsules is also good (>50%). Lipophilic prodrugs of F-ddI activated by adenosine deaminase also continue under investigation. HPLC methodology for the rapid measurement of F-ddI progrugs such as 6- chloro-2',3'-dideoxypurineriboside and its metabolites in biological fluids and tissues is being developed. Direct fluorogenic derivatization of cellular extracts in conjunction with paired-ion HPLC is being employed for the nonradiochemical measurement of subpicomole amounts of intracellular F-ddATP, the active metabolite of both F-ddA and F-ddI. F-ddATP levels will be measured in periperal blood mononuclear cells from patients treated with F-ddA and correlated with observed anti-HIV activity.