Pharmacology of HIV Viral DNA & Retroviral Integrases

为了进一步扩大开发抗逆转录病毒药物的靶点数量，我们正在使用重组酶和对应于前病毒末端的短寡核苷酸（LTR）进行体外测定，研究HIV-1整合酶抑制剂。整合酶是抑制剂开发的基本靶点，因为它对病毒复制至关重要。它也由HIV编码，没有细胞等价物。我们的实验室开创了这一研究领域，并报告了各种抑制剂家族。 我们继续与Burke博士一起研究多羟基芳烃。因为其中之一，咖啡酰奎尼酸是抗病毒的，我们进行了菊苣酸类似物的结构-活性研究。我们发现，通过转化为四乙酸酯来阻断儿茶酚官能团几乎不会导致效力损失，这取决于中心接头上存在至少一个羧基。作为一个整体，我们的工作导致了新的整合酶抑制剂，这可能被视为具有抗病毒活性的双咖啡酰衍生物的鉴定。 我们还发现了一个新的HIV整合酶抑制剂家族，噻唑硫氮杂卓是非儿茶酚抑制剂。值得注意的是，这些硫代硫氮杂卓衍生物在镁（被认为是生理二价阳离子）以及基于镁的测定（其在体外更稳健）中具有活性。它们是抗病毒的，不抑制除整合酶以外的其他逆转录病毒靶点。需要进一步的工作来确定整合酶是否是这类新药的唯一体内靶点。 我们还在研究新型抑制剂，它们可以通过与前病毒DNA末端结合来阻止整合，以及小肽和核苷酸抑制剂。

In an effort to further extend the number of targets for development of anti-retroviral agents, we are studying HIV-1 integrase inhibitors using in vitro assays using recombinant enzyme and short oligonucleotides corresponding to the proviral ends (LTR's). Integrase is a rationale target for inhibitor development because it is essential for viral replication. It is also encoded by HIV and does not have a cellular equivalent. Our laboratory has pioneered this research field and reported various families of inhibitors. We have continued our studies on polyhydroxylated aromatics with Dr. Burke. Because one of them, caffeoylquinic acid is antiviral, we performed structure-activity studies on chicoric acid analogs. We found that blocking the catechol functionality through conversion to tetraacetate esters results in almost no loss of potency, contingent of the presence of at least one carboxyl group on the central linker. Taken as a whole, our work has resulted in the identification of new integrase inhibitors, which may be regarded as bis-caffeoyl derivatives with antiviral activity. We also discovered a novel family of HIV integrase inhibitors, the thiazolothiazepine that are non-catechol inhibitors. Remarkably, these thiolothiazepine derivatives are active in magnesium (considered to be the physiological divalent cation) as well as in maganese-based assays (which are more robust in vitro). They are antiviral without inhibiting other retroviral targets besides integrase. Further work will be needed to determine whether integrase is the only in vivo target of this new class of drugs. We are also studying novel types of inhibitors that can prevent integration by binding to the proviral DNA ends as well as small peptide and nucleotide inhibitors.