THE ANALYTICAL CHEMISTRY OF ANTI-AIDS AGENTS

该项目的目的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药物和新型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、化学和生化性质，包括辛醇-水分配系数；(3)在生物样品中测定这些药物及其代谢物，以阐明药理学和确定药代动力学。高效液相色谱法和质谱法是重点技术。II期药物2-b-氟-2,3-二脱氧腺苷（F-ddA, lodenosine）及其脱胺抗hiv活性代谢物2-b-氟-2,3-二脱氧腺苷（F-ddI）仍然是主要感兴趣的化合物。采用反相高效液相色谱法验证的分析策略被应用于常规和超灵敏测量hiv感染的人类生物体液中的F-ddA。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物在成年艾滋病患者中的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个组成部分，与单药治疗的F-ddA完全具有生物等效性。细胞提取物的直接荧光衍生化结合对离子高效液相色谱法被用于细胞内低皮摩尔量的F-ddATP （F-ddA和F-ddI的活性代谢物）的非放射化学测量。F-ddATP水平在接受F-ddA治疗的患者外周血单个核细胞中被测量，并将与观察到的抗hiv活性相关。由腺苷脱氨酶（ADA）激活的F-ddI亲脂性前药也在继续研究中，作为治疗隐匿在中枢神经系统（CNS）的HIV的潜在药物。生物体液和组织中6-氯-2,3-二脱氧嘌呤内苷及其代谢产物的高效液相色谱快速测定方法正在开发和应用。以F-ddA为模型化合物，建立了ada活化F-ddI前药的生理药动学模型。该模型被用于研究各种生理生化过程的影响，重点是药物前胃肠道吸收、血脑屏障渗透到中枢神经系统和代谢激活。在大鼠、猴子和人类中静脉注射和口服F- ddA后，血浆和大脑中的浓度与时间的关系数据被用于模型验证和物种间缩放。题目：抗艾滋病药物的分析化学-腺苷脱氨酶，艾滋病，荧光衍生化，液相色谱，药代动力学，前药，2'- β -氟-2',3'-二脱氧腺苷，-人体组织，液体，细胞等。

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 are being applied 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 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 is being 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 levels are being measured in periperal blood mononuclear cells from patients treated with F-ddA and will be correlated with observed anti-HIV activity. 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). HPLC methodology for the rapid measurement of F-ddI progrugs, such as 6- chloro-2,3-dideoxypurineriboside, and their metabolites in biological fluids and tissues is being developed and applied. 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 - adenosine deaminase, AIDS, fluorogenic derivatization, liquid chromatography, pharmacokinetics, prodrugs, 2'-beta-fluoro-2',3'- dideoxyadenosine, - Human Tissues, Fluids, Cells, etc.