Functional and inhibitory studies of human lipoxygenase

描述（由申请人提供）：本研究计划的范围是扩展原始R01的Aim 3的一小部分，其目标有限。我们目前的修订旨在发现100多种脂氧合酶抑制剂，优化它们的活性并提供足够数量的抑制剂，以研究它们作为治疗癌症、1型糖尿病和帕金森病的药物。这项研究将非常费力，需要在全国四个地点增加人员和设备——这是刺激计划的关键理由。人脂氧合酶（hLO）同工酶是潜在的治疗靶点，因为它们与许多人类疾病有关，但由于难以靶向特异性的hLO同工酶，其开发进展很少。我们建议通过发现针对四种hLOs、5-hLO、血小板12-hLO、网状细胞15-hLO-1和表皮15-hLO-2的特异性抑制剂来纠正这一问题，并确定它们对人类疾病的疗效。这项拨款有四个主要目的。第一个主要目标是从我们最初的高通量NIH发现的超过1000种抑制剂中确定所有可能的同工酶特异性脂氧合酶抑制剂。随后，将对约100种预期确定的抑制剂中的每一种进行结构修饰，以优化其效力/选择性。第二个主要目的是确定这些抑制剂是否对癌症具有强效/选择性。目前，我们的初步结果表明，我们的一种5-hLO抑制剂对小鼠结肠癌具有强效/选择性，我们计划对我们的一系列癌细胞系筛选剩余的抑制剂。第三个主要目标是确定我们的12-hLO抑制剂是否保护2细胞免受自身免疫破坏，从而预防1型糖尿病。目前，我们有初步的结果表明，我们的12-hLO抑制剂可以降低IL-12的浓度，表明这种抑制剂可以保护2-细胞免于死亡。我们建议扩展这一结果，并测试我们的抑制剂对非肥胖糖尿病（NOD）小鼠的功效。第四个主要目标是确定我们的LO抑制剂是否保护神经细胞免受帕金森氏病中发生的炎症损伤。目前，我们有初步的结果表明，我们的抑制剂进入神经细胞并保护它们免受谷氨酸诱导的细胞凋亡。我们现在正在研究哪种特定的LO抑制剂对我们的帕金森小鼠模型最有益。这些都是非常令人鼓舞的结果，并构成了一个特定的LO抑制剂“工具箱”，这将使我们能够加快了解LO在癌症，糖尿病和帕金森病中的作用。

DESCRIPTION (provided by applicant): The scope of this research program is to expand on a small part of Aim 3 of the original R01, which had limited goals. Our current revision aims to discover over 100 lipoxygenase inhibitors, optimize their activity and provide sufficient quanitites of inhibitor to investigate them as therapeutics against cancer, type-1 diabetes and Parkinson's disease. This research will be very laborious and require additional staff and equipment in four locations across the nation- key rationales of the stimulus package. Human lipoxygenase (hLO) isozymes are potential therapeutic targets because they are involved in numerous human diseases and yet, there has been little progress in their development due to the difficulty of targeting specific hLO isozymes. We propose to rectify this problem by discovering specific inhibitors against four hLOs, 5-hLO, platelet 12-hLO, reticulocyte 15-hLO-1 and epidermal 15-hLO-2, and determining their efficacy against human diseases. There are four major aims of the grant. The first major aim is to identify all of the possible isozyme specific lipoxygenase inhibitors from our initial high through-put NIH discovery of over 1000 inhibitors. Subsequently, each of the ~100 anticipated identified inhibitors will be structurally modified to optimize their potency/selectivity. The second major aim is to determine if these inhibitors are potent/selective against cancer. Currently, we have preliminary results which demonstrate that one of our 5-hLO inhibitors is potent/selective against murine colon cancer and we plan to screen the remaining inhibitors against our suite of cancer cell lines. The third major aim is to determine if our 12-hLO inhibitors protect 2-cells from auto-immune destruction and hence protect against type-1 diabetes. Currently, we have preliminary results which show that our 12-hLO inhibitor lowers the concentration of IL-12, indicating that this inhibitor may protect 2 -cells from death. We propose to expand this result and test the efficacy of our inhibitors on non-obese diabetic (NOD) mice. The fourth major aim is to determine if our LO inhibitors protect neuronal cells from the inflammatory damage that occurs in Parkinson's disease. Currently, we have preliminary results which demonstrate that our inhibitors enter neuronal cells and protect them from glutamate induced apoptosis. We are now investigating which of the specific LO inhibitors are the most beneficial in our Parkinson's mouse model. These are very encouraging results and constitute a specific LO inhibitor "tool box" which will allow us to accelerate the tempo in understanding the role of LO in cancer, diabetes and Parkinson's disease. PUBLIC HEALTH RELEVANCE: Lipoxygenase (LO) is a critical enzyme involved in numerous human diseases. The goal of this proposal is to discover and characterize inhibitors to LO with the goal of developing possible therapeutics against cancer, type-1 diabetes and Parkinson's disease.