Functional and inhibitory studies of human lipoxygenase

描述(申请人提供)：本研究计划的范围是对最初的R01的目标3的一小部分进行扩展，该部分的目标有限。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来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工作人员和设备--这是刺激方案的关键理由。人类脂氧合酶(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，但由于难以靶向特定的HLO同工酶，其研究进展甚微。我们建议通过发现针对4种HLO、5-HLO、血小板12-HLO、网织红细胞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。这笔赠款有四个主要目的。第一个主要目标是从我们最初发现的1000多种高通量NIH抑制剂中识别所有可能的同工酶特异性脂氧合酶抑制剂。随后，将对~100种预期确定的抑制剂进行结构修饰，以优化它们的效力/选择性。第二个主要目标是确定这些抑制物对癌症是否有效/选择性。目前，我们的初步结果表明，我们的其中一种5-HLO抑制剂对小鼠结肠癌是有效的/选择性的，我们计划筛选其余的抑制剂来对抗我们的癌细胞系。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，我们已有的初步结果表明，我们的12-HLO抑制剂降低了IL-12的浓度，表明该抑制剂可能保护2-细胞免于死亡。我们建议扩大这一结果，并在非肥胖糖尿病(NOD)小鼠身上测试我们的抑制剂的有效性。第四个主要目标是确定我们的LO抑制剂是否能保护神经细胞免受帕金森氏病所致的炎性损伤。目前，我们有初步的结果表明，我们的抑制剂进入神经细胞，并保护他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抑制剂在我们的帕金森氏症小鼠模型中最有益。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”，将使我们能够加快节奏，了解LO在癌症、糖尿病和帕金森病中的作用。 与公共卫生相关：脂氧合酶(LO)是一种与多种人类疾病有关的关键酶。这项建议的目标是发现和表征LO的抑制剂，目标是开发可能的治疗癌症、1型糖尿病和帕金森病的药物。

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.