人网织红细胞12/15-脂氧合酶（h12/15-LOX）是一种脂质氧化酶，可以在没有磷脂酶的情况下直接氧化脂质膜，从而直接攻击线粒体等细胞器。h12/15-LOX 的这种细胞毒性活性在中风后在神经元和内皮细胞中上调，并被认为导致神经元细胞死亡和血脑屏障渗漏。体外选择性靶向重组 h12/15-LOX 并具有离体小鼠直系同源活性的抑制剂的发现，可能会支持一种治疗中风的新治疗策略。在此，我们报告了在高通量筛选 (HTS) 中发现的一个新的抑制剂家族，该家族对重组 h12/15-LOX 和细胞小鼠 12/15-LOX (m12/15-LOX) 具有选择性且有效。MLS000099089（化合物99089）是母体分子，在体外针对 h12/15-LOX 表现出 3.4 ± 0.5 μM 的IC 50效力，在小鼠神经元细胞系 HT-22 中表现出约 10 μM 的离体 IC 50效力。当在 20 μM 抑制剂浓度下进行测试时，化合物99089与 h5-LOX 和 COX-2 相比，选择性高出 30 倍以上，与 h15-LOX-2 相比，选择性高出 15 倍，与 h12-LOX 相比，选择性高出 10 倍。稳态抑制动力学表明，99089对 h12/15-LOX 的抑制模式是混合抑制剂的抑制模式，K ic为 1.0 ± 0.08 μM，K iu为 6.0 ± 3.3 μM。这些数据表明，99089和相关衍生物可以作为抗中风疗法开发的起点，因为它们能够在体外选择性靶向 h12/15-LOX 和离体靶向 m12/15-LOX。

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Human reticulocyte 12/15-lipoxygenase (h12/15-LOX) is a lipid-oxidizing enzyme that can directly oxidize lipid membranes in the absence of a phospholipase, leading to a direct attack on organelles, such as the mitochondria. This cytotoxic activity of h12/15-LOX is up-regulated in neurons and endothelial cells after a stroke and thought to contribute to both neuronal cell death and blood–brain barrier leakage. The discovery of inhibitors that selectively target recombinant h12/15-LOX in vitro, as well as possessing activity against the murine ortholog ex vivo, could potentially support a novel therapeutic strategy for the treatment of stroke. Herein, we report a new family of inhibitors discovered in a High Throughput Screen (HTS) that are selective and potent against recombinant h12/15-LOX and cellular mouse 12/15-LOX (m12/15-LOX). MLS000099089 (compound 99089), the parent molecule, exhibits an IC50 potency of 3.4 ± 0.5 μM against h12/15-LOX in vitro and an ex vivo IC50 potency of approximately 10 μM in a mouse neuronal cell line, HT-22. Compound 99089 displays greater than 30-fold selectivity versus h5-LOX and COX-2, 15-fold versus h15-LOX-2 and 10-fold versus h12-LOX, when tested at 20 μM inhibitor concentration. Steady-state inhibition kinetics reveals that the mode of inhibition of 99089 against h12/15-LOX is that of a mixed inhibitor with a Kic of 1.0 ± 0.08 μM and a Kiu of 6.0 ± 3.3 μM. These data indicate that 99089 and related derivatives may serve as a starting point for the development of anti-stroke therapeutics due to their ability to selectively target h12/15-LOX in vitro and m12/15-LOX ex vivo.