Pharmacology of HIV Viral DNA & Retroviral Integrases

为了进一步扩大抗逆转录病毒药物开发的靶点，我们正在利用重组酶和与前病毒末端(LTR‘s)相对应的短寡核苷酸，在体外试验中研究HIV-1整合酶抑制剂。整合酶是开发抑制剂的基本靶点，因为它对病毒复制是必不可少的。它也是由艾滋病毒编码的，没有细胞上的等价物。我们的实验室开创了这一研究领域，报道了不同家族的缓蚀剂。我们与伯克博士一起继续我们对多羟基芳烃的研究。因为其中一种咖啡酰奎宁酸是抗病毒的，所以我们进行了菊菊酸类似物的构效研究。我们发现，通过转化为四乙酸酯来阻断儿茶酚的官能团几乎不会导致效力的损失，这取决于中心连接基上至少存在一个羧基。总体而言，我们的工作导致了新的整合酶抑制剂的鉴定，这可能是具有抗病毒活性的双咖啡酰基衍生物。我们还发现了一类新的HIV整合酶抑制剂家族，噻唑并氮卓类非儿茶酚类抑制剂。值得注意的是，这些硫代硫杂卓衍生物在镁(被认为是生理上的二价阳离子)以及基于锰的分析(在体外更可靠)中都是有效的。它们是抗病毒的，但不抑制整合酶以外的其他逆转录病毒靶标。还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，这些抑制剂可以通过与前病毒DNA末端以及小肽和核苷酸抑制剂结合来防止整合。

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.