Sinko William, Sierecki Emma, de Oliveira César A F, McCammon J Andrew
Department of Chemistry and Biochemistry, Center for Theoretical Biological Physics, University of California, San Diego, La Jolla, CA, USA.
Methods Mol Biol. 2012;819:561-73. doi: 10.1007/978-1-61779-465-0_33.
We present an example-based description of virtual screening (VS) techniques used to identify new regulators of the Akt phosphatase PHLPP (PH domain Leucine repeat Protein Phosphatase). This enzyme opposes the effects of two kinases, Akt and PKC, which play a major role in cell growth and survival. Therefore, PHLPP is a potential therapeutic target in pathophysiologies where these pathways are either repressed, such as in diabetes and cardiovascular diseases, or over-activated as in cancer. To the best of our knowledge, no PHLPP inhibitors have been reported so far in the literature. In this study, we used a combination of chemical and virtual screening techniques that led to the identification of a number of inhibiting compounds with diverse scaffolds. These compounds bind PHLPP and inhibit cell death when tested in cellular assays. We employed GLIDE docking software to screen a library of more than 40,000 compounds selected from the NCI open depository (250,000 compounds) by similarity searches. We compare the efficiency at which we determined binding compounds from the chemical screen, and compare enrichment factors of the virtually discovered compounds over chemical screening.
我们展示了一个基于实例的虚拟筛选（VS）技术描述，该技术用于识别Akt磷酸酶PHLPP（PH结构域亮氨酸重复蛋白磷酸酶）的新调节剂。这种酶对抗两种激酶Akt和PKC的作用，这两种激酶在细胞生长和存活中起主要作用。因此，在这些信号通路被抑制的病理生理学中，如糖尿病和心血管疾病，或者在癌症中过度激活时，PHLPP是一个潜在的治疗靶点。据我们所知，目前文献中尚未报道过PHLPP抑制剂。在本研究中，我们使用了化学和虚拟筛选技术的组合，从而鉴定出了许多具有不同骨架的抑制性化合物。这些化合物在细胞试验中测试时能结合PHLPP并抑制细胞死亡。我们使用GLIDE对接软件筛选了一个通过相似性搜索从美国国立癌症研究所开放库（250,000种化合物）中选出的超过40,000种化合物的文库。我们比较了从化学筛选中确定结合化合物的效率，并比较了虚拟发现的化合物相对于化学筛选的富集因子。
