MOLECULAR PROBES OF THE MECHANISM OF CYTOCHROME P450

这一计划的中心主题是研究脑出血的机制 细胞色素P450酶和一氧化氮合酶的作用。这个 主要途径是活性物质的合成和表征。 铁的卟啉物种，被认为是 催化循环，并联系这些物种的相互转化 对这些蛋白质的分子理解。细胞色素P450是 参与药物解毒和激素的中心蛋白质 代谢的同时，一氧化氮和过氧亚硝酸盐。合成的 可以使用金属卟啉作为探针来干预这些 以诊断方式进行处理。因此，这些代理人可能会被证明是 阐述超氧化物生物学的重要工具， 过氧亚硝酸盐和NO。同样是这些金属卟啉已经显示出令人印象深刻 动物体内活性提示其作为药物的应用 探员们。我们的努力试图提供一个机械性和 可应用于体外模型、细胞的动力学信息 特定疾病状态的培养研究和整体动物模型 如缺血再灌注、脓毒症和自身免疫性疾病。实验是 旨在确定形成什么活性中间体和什么 他们的生物目标很可能是。对这些遗嘱的阐述 工艺将促进金属络合物的设计 催化分解过亚硝酸根和这些其他物种。这个 一氧化氮合酶的研究旨在阐明 一氧化氮合酶的作用机制和寻找可能有助于氧化的新过程 随着一氧化氮合酶抑制剂的合理开发。过氧铁(III) 现在，在水溶液中很容易接触到卟啉物种 环境温度。快速动力学技术已经发展到 研究这些物种的反应能力。一个中心问题是如何 调节亲核和亲核之间这些络合物的化学 亲电通路对于理解细胞的多样性至关重要 P450介导的过程。合成和半合成磷脂 程序集用于模拟和理解中的较大规模的事件 P450和NOS的作用及膜通透性的探讨 活性氧物种。两亲性膜结合 金属卟啉对pH特别敏感。这是一个 史无前例的观察，可能会立即应用于 金属药品的设计。

The central theme of this program involves studies of the mechanisms of action of cytochrome P450 enzymes and nitric oxide synthase. The principal approach is the synthesis and characterization of reactive iron porphyrin species that are thought to be intermediates in the catalytic cycles and to relate the interconversions of these species toward a molecular understanding of these proteins. Cytochrome P450 is the central protein involved in drug detoxification and hormone metabolism while nitric oxide and peroxynitrite. Synthetic metalloporphyrins can be employed as probes to intervene in these processes in diagnostic ways. Thus, these agents may prove to be significant tools for elaborating the biology of superoxide, peroxynitrite and NO. These same metalloporphyrins have shown impressive activity in animals suggesting their application as pharmaceutical agents. Our effort seeks to provide a foundation of mechanistic and kinetic information which can be applied to in vitro models, cell culture studies and whole animal models of specific disease states such as ischemia-reperfusion, sepsis and autoimmune diseases. Experiments are aimed at determining what reactive intermediates are formed and what their biological targets are likely to be. The elaboration of these will processes will facilitate the design of metal complexes for the catalytic decomposition of peroxynitrite and these other species. The studies of nitric oxide synthase aim to illuminate the range of mechanisms of NOS and to seek out new oxidation processes that may help with the rational development of NOS inhibitors. Peroxoiron(III) porphyrin species are now readily accessible in aqueous solution at ambient temperature. Rapid kinetic techniques have been developed to study the reactivity of these species. A central question is how to modulate the chemistry of these complexes between nucleophilic and electrophilic pathways that are essential to understand the variety of P450 mediated processes. Synthetic and semi-synthetic phospholipid assemblies are used to model and understand the larger scale events in P450 and NOS action and to probe the permeability of membranes to reactive oxygen species. Membrane binding by amphiphilic metalloporphyrins is extraordinarily sensitive to pH. This is an unprecedented observation that may have immediate application to the design of metallopharmaceuticals.