High Throughput and Virtual Screening for Human 12-LO, 15-LO-1, and 15-LO-2 Inhib

人类 12-LO、15-LO-1 和 15-LO-2 抑制物的高通量和虚拟筛选

• 批准号: 7368412
• 负责人: Theodore R Holman
• 金额: $ 2.5万
• 依托单位: UNIVERSITY OF CALIFORNIA SANTA CRUZ
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-31 至 2009-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7368412
• 关键词: Animals; Arachidonate 12-Lipoxygenase; Arachidonate 15-Lipoxygenase; Arachidonate 5-Lipoxygenase; Arachidonic Acids; Binding; Biological Assay; Biological Factors; Biology; Blood Platelets; Cell Line; Cells; Chemicals; Collaborations; Collection; Computer Analysis; Computer Simulation; Data; Disease; Docking; Enzymes; Flavonoids; Genomics; Germination; Grant; Homology Modeling; Hormones; Housing; Human; Inflammatory; Inhibitory Concentration 50; Ischemia; Isoenzymes; Kinetics; Lead; Leukotrienes; Libraries; Lipoxins; Lipoxygenase; Lipoxygenase 1; Lipoxygenase 2; Lipoxygenase Inhibitors; Malignant Neoplasms; Malignant neoplasm of prostate; Manuals; Methods; Modeling; Neurons; Numbers; Organism; Plants; Play; Rate; Reticulocytes; Robot; Role; Scientist; Screening Result; Screening procedure; Signaling Molecule; Specificity; Technology; Therapeutic; United States National Institutes of Health; Work; caN protocol; cancer cell; cost; design; drug discovery; high throughput screening; human disease; human tissue; improved; in vivo; inhibitor/antagonist; novel; repository; senescence; small molecule; virtual

DESCRIPTION (provided by applicant): High-throughput screening (HTS) is a major method in early drug discovery and has yet to be fully applied to all human lipoxygenase (hLO) isozymes, critical enzymes in the progression of a number of human diseases. We have currently discovered over 20 novel inhibitors against human lipoxygenase primarily utilizing manual screening but have recently developed a high-throughput assay to increase the rate of discovery. We would now like to apply this HTS technology to the approximately 73,000 compounds at the NIH DPISMR to identify specific inhibitors to each of our three LO isozymes (12- hLO, 15-hLO-1 and 15-hLO-2). The specific inhibitors we discover will then be screened against specific cell lines to determine their in-vivo activity against prostate cancer cells and neuronal cells (as ischemia models). In parallel with the HTS, and at no extra cost to the NIH, we will dock the full HTS library against our three lipoxygenase homology models (12-hLO, 15-hLO-1 and 15-hLO-2) and compare the docking hits to those found by the HTS assay. This comparison will allow us to both improve our in silico methods and help us structurally interpret the HTS data we will obtain. Currently, we have shown both the accuracy and the reliability of this screen against a 47 compound flavonoid library and the NCI 3104 compound Mech./Div./Nat.Prod. library. In addition, Anton Simeonov, at NIH Chemical Genomics Center (NCGC), has successfully screened the LOPAC library with the Kalypsys robot and shown known LO inhibitors can be accurately found. Finally, modeling predictions with Prof. Matt Jacobson have already produced inhibitors from earlier screens. All of this data suggests that these aims are not only feasible but highly likely to be successful. Lipoxygenases (LO) are widely distributed throughout the plant and animal kingdoms and play a central role in the biology of these organisms. In plants they are involved in germination and senescence. In human tissue, there are three major human lipoxygenases, 5-, 12-, and 15-LO, whose primary enzymatic difference lies in their positional specificity on arachidonic acid (AA). The products of lipoxygenase are the precursors of hormones, such as leukotrienes and lipoxins, which have been implicated as critical signaling molecules in a variety of inflammatory diseases and cancers. There is currently strong evidence that if scientists can discover potent inhibitors against each specific lipoxygenase isozyme, these may become effective cellular probes and therapeutics against LO. This grant submission proposes to do exactly this for platlet 12-LO, reticulocyte 15-LO-1 and epidermal 15- LO-2.

描述（由申请人提供）：高通量筛选（HTS）是早期药物发现的主要方法，尚未完全应用于所有人类脂氧酶（HLO）同工酶，这是许多人类疾病的进展中的关键酶。我们目前已经发现了20多种针对人脂氧酶的新型抑制剂主要利用手动筛选，但最近开发了一种高通量测定法以提高发现率。现在，我们想将这种HTS技术应用于NIH DPISMR的大约73,000种化合物，以鉴定三个LO同工酶中的每一种（12- HLO，15-HLO-1和15-HLO-2）的特定抑制剂。然后，我们发现的特定抑制剂将被筛选针对特定的细胞系，以确定其对前列腺癌细胞和神经元细胞的体内活性（作为缺血模型）。与HTS同时，NIH无需额外费用，我们将与三个Lipoxygoganase同源性模型（12-HLO，15-HLO-1和15-HLO-2）进行对接，并将对接命中与HTS Assay发现的命中进行比较。这种比较将使我们既可以改善我们的硅方法，又可以帮助我们在结构上解释我们将获得的HTS数据。当前，我们已经显示了此屏幕相对于47复合类黄酮库和NCI 3104复合Mech./div./nat.prod的精度和可靠性。图书馆。此外，NIH化学基因组学中心（NCGC）的Anton Simeonov已成功地使用Kalypsys机器人筛选了LOPAC文库，并可以准确地找到已知的LO抑制剂。最后，用马特·雅各布森（Matt Jacobson）教授对预测进行建模已经产生了早期筛选的抑制剂。所有这些数据都表明，这些目标不仅是可行的，而且很可能成功。脂氧酶（LO）广泛分布在整个动植物界，并在这些生物的生物学中起着核心作用。在植物中，它们参与发芽和衰老。在人体组织中，有三种主要的人脂氧酶，5-，12-和15-lo，其主要酶促差在于它们对花生四烯酸（AA）的位置特异性。脂氧酶的产物是激素的前体，例如白细胞和lipoxins，它们被认为是多种炎性疾病和癌症中的关键信号分子。目前有强有力的证据表明，如果科学家能够发现针对每个特定的脂氧合酶同工酶的有效抑制剂，则可能成为有效的细胞探针和对LO的治疗剂。该赠款提交的建议是针对Platlet 12-LO，网状细胞15-LO-1和表皮15-LO-2的精确执行此操作。