HIV RNase H Natural Product Inhibitors: Isolation and Optimization

我们的目标是鉴定和优化植物来源的小分子，用于抑制HIV RT相关的 RNase H（RNH）活性，HIV干预的新靶点。本项目的总体目标是推荐 2-3来自10，000个级分的初始筛选的用于开发的化合物（CD）（第一程序）和 推荐4-6张CD（备份程序）从筛选其余的植物来源文库（150，000 分数）。第一代CD应达到50 nm或更好的IC 50，而第二代CD应达到50 nm或更好的IC 50。 应具有个位数nm IC 50。这些化合物应该能有效抑制HIV病毒株， 对现有药物有抵抗力。RNH抑制剂（RNHI）将使用生物化学和病毒学数据进行优化 首先利用从X射线晶体学获得的计算和结构信息， RT-RNHI复合物的研究（见本RFA中的其他项目），以允许合理设计类似物， 将是人工合成或半人工合成的。如果采用半合成方法，我们可以使用 我们的50 L生物反应器生产克数量的起始材料，使我们能够产生一个小的文库， 的类比。此外，组合和平行合成将用于一些分子，以增加 潜在线索的数量和化学多样性我们将坚持既定的指导方针，如 Lipinski的“五个规则”，以增加我们获得药物样分子的机会。类比显示 将对活性改善的药物进行ADME/PK研究，以选择药物样先导物。这一进程将 直到我们获得表4中描述的CD的轮廓。我们还想加入另一个 在RNHI的药物发现中的方法。我们会利用一些线索的代谢产物 作为鉴定RNHI的新化学型的来源。我们对结果感到非常鼓舞， 到目前为止（表1）。生化值和基于细胞的值之间有很强的相关性，一些 IC 50为<1uM with an excellent therapeutic Index >200。这是在以下方面向前迈出的重要一步： 识别和开发RNHI。这是对我们迄今为止开发的植物细胞培养技术的致敬。 一个高度多样化的图书馆，来自2200多个植物物种，代表284个植物科， 其特征在于次级代谢产物生产的高再现性。这个项目有巨大的 在提供一类新的艾滋病毒药物方面的潜在影响。

Our objective is to identify and optimize plant derived small molecules for the inhibition of HIV RT-associated RNase H (RNH) avtivity, a novel target for HIV intervention. The overall goal of this poject is to recommend 2-3 compounds for development (CD) from the initial screen of 10,000 fractions (1st program) and recommend 4-6 CDs (back up program) from screening the rest of the plant derived library (150,000 fractions). The first generation CDs should attain an IC50 of 50 nm or better while second generation CDs should have single digit nm IC50. These compounds should be effective in ihibiting HIV strains that are resistant to current drugs. RNH inhibitors (RNHI) will be optimized using biochemical and virologic data initially followed by utilizing computanional and structural information obtained from X-ray crystallographic studies of RT-RNHI complexes (see other projects within this RFA) to allow rational design of analogs that will be either synthesized or semi-synthesized. In case a semi-synthetic approach is adopted, we can use our 50 L bioreactor to produce gram quantities of the starting material to allow us to generate a small library of analogs. In addition, combinatorial and parallel synthesis will be utilized for some molecules to increase both the quantity and chemical diversity of potential leads. We will adhere to established guidlines such as the Lipinski "Rule of five" to increase our chances of obtaining drug like molecules. Analogs that show improved acivity will be subjected to ADME/PK studies in order to select for drug like leads. This process will be repeated until we achieve the profile of a CD described in Table 4. We would like to include yet another approach in our drug discovery of RNHI. We will exploit the generation of metabolites of some of our leads as a source of new chemotypes for identifyiing RNHI. We are very encouraged by the results that we obtained so far (Tablel). There is a strong correlation between the biochemical and cell-based values, some having an IC50<1uM with an excellent therapeutic Index >200. This represents a major step forward in identifying and developing RNHIs. It is a tribute to the plant cell culture technology that we developed so far. A highly diverse library that originated from more than 2200 plant species, representing 284 plant families, that is charecarized by a high reproducibility of secondary metabolite production. This project has enormous potential impact in delivering a new class of HIV drugs.