The Analytical Chemistry of Anti-AIDS Agents

本项目的目标是研究和开发合适的生物分析方法，以：（1）确定潜在的抗艾滋病剂和新的抗病毒药物的结构和纯度，（2）确定这些化合物及其代谢物的物理、化学和生物化学性质，包括辛醇-水分配系数，以及（3）测量生物样品中的这些药物及其代谢物以阐明药理学并确定药代动力学。高效液相色谱（HPLC）和质谱是重点技术。II期药物2 '-b-氟-2'，3 '-双脱氧腺苷（F-ddA，碘腺苷）及其脱氨基抗HIV活性代谢物2'-b-氟-2 '，3'-双脱氧肌苷（F-ddI）仍然是主要感兴趣的化合物。采用反相HPLC的验证分析策略已被用于HIV感染的人类生物体液中F-ddA的常规和超灵敏测量。 口服F-ddA胶囊和液体制剂的人体代谢、分布和药代动力学已与该药剂在成人AIDS患者中的两部分I期临床试验一起确定。口服F-ddA作为联合抗逆转录病毒治疗的一部分与F-ddA作为单药治疗完全生物等效。细胞提取物的直接荧光衍生化结合双离子HPLC已被用于低皮摩尔量的细胞内F-ddATP，F-ddA和F-ddI的活性代谢产物的非放射化学测量。F-ddATP可以在接受F-ddA治疗的患者的外周血单核细胞中测量，但没有足够的数据与观察到的抗HIV活性相关。 正在研究2 '-氟-2-脱氧腺苷的毒性、代谢和生化药理学，以确定该微量成分在碘腺苷毒性特征中的作用。由腺苷脱氨酶（ADA）激活的F-ddI的亲脂性前药也继续作为治疗中枢神经系统（CNS）中隔离的HIV的潜在药物进行研究。 以F-ddA为模型化合物，建立了生理药代动力学模型，研究了ADA激活的F-ddI前体药物的处置。该模型被用于研究各种生理和生化过程的影响，重点是前药胃肠道吸收，血脑屏障渗透到CNS和代谢活化。 在大鼠、猴和人类中静脉和经口给予F-ddA后，血浆和脑中的浓度与时间数据正用于模型验证和种间缩放。 抗艾滋病药物的分析化学

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 II drug 2'-b-fluoro-2',3'-dideoxyadenosine (F-ddA, lodenosine) and its deaminated anti-HIV-active metabolite 2'-b-fluoro-2',3'-dideoxyinosine (F-ddI) remain the compounds of primary interest. Validated analytical strategies employing reversed-phase HPLC have been used for both the routine and ultrasensitive measurement of F-ddA in HIV-infected human biological fluids. The human metabolism, distribution and pharmacokinetics of oral F-ddA, in both capsule and liquid formulation, have been determined in conjunction with a two-part Phase I clinical trial of this agent in adult AIDS patients. Oral F-ddA given as a component of combination antiretroviral therapy is fully bioequivalent with F-ddA given as monotherapy. Direct fluorogenic derivatization of cellular extracts in conjunction with paired-ion HPLC has been employed for the nonradiochemical measurement of low picomole amounts of intracellular F-ddATP, the active metabolite of both F-ddA and F-ddI. F-ddATP can be measured in periperal blood mononuclear cells from patients treated with F-ddA, but sufficient data is not available to correlate with observed anti-HIV activity. The toxicity, metabolism and biochemical pharmacology of 2'-fluoro-2-deoxyadenosine are under investigation to determine the role of this trace constituent in the toxicity profile of lodenosine. Lipophilic prodrugs of F-ddI activated by adenosine deaminase (ADA) also continue under investigation as potential agents for the treatment of HIV sequestered in the central nervous system (CNS). A physiological pharmacokinetic model has been constructed to study the disposition of selected ADA-activated F-ddI prodrugs by using F-ddA as a model compound. This model is being used to investigate the effects of various physiological and biochemical processes with emphasis on prodrug gastrointestinal absorption, blood-brain-barrier penetration into the CNS, and metabolic activation. Concentration versus time data in plasma and brain following intravenous and oral administration of F-ddA in rats, monkeys and humans are being used for model validation and interspecies scaling. AIDS Title: The Analytical Chemistry of Anti-AIDS Agents