BRP-187: A potent inhibitor of leukotriene biosynthesis that acts through impeding the dynamic 5-lipoxygenase/5-lipoxygenase-activating protein (FLAP) complex assembly
BRP-187: A potent inhibitor of leukotriene biosynthesis that acts through impeding the dynamic 5-lipoxygenase/5-lipoxygenase-activating protein (FLAP) complex assembly
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DOI：
10.1016/j.bcp.2016.08.023
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发表时间：
2016-11-01
影响因子：
5.8
通讯作者：
Werz, Oliver
中科院分区：
文献类型：
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作者：
Garscha, Ulrike;Voelker, Susanna;Werz, Oliver
促炎性白三烯（LT）由活化的白细胞中的花生四烯酸（AA）形成，其中5-脂氧合酶（5-LO）易位至核膜以与整合的核膜蛋白5-LO-活化蛋白（FLAP）组装功能复合物。FLAP是MAPEG家族的成员之一，它促进AA向5-LO的转化，而LT的生物合成主要依赖于FLAP。在这里，我们表明，新的LT生物合成抑制剂BRP-187阻止5-LO/FLAP在人白细胞核膜的相互作用，而不阻断5-LO核再分配。BRP-187抑制脂多糖加N-甲酰基-甲硫氨酰基-亮氨酰基-苯丙氨酸刺激的人单核细胞和多形核白细胞中的5-LO产物形成（IC 50 = 7-10 nM），以及离子载体A23187激活后的5-LO产物形成（IC 50 = 10-60 nM）。过量的外源性AA显著损害BRP-187的效力。无细胞测定中的直接5-LO抑制仅在>35倍高的浓度下是明显的，这是可逆的并且在还原条件下没有改善。BRP-187阻止A23187诱导的5-LO/FLAP复合物在白细胞中的组装，但未能阻断5-LO核转位，这与FLAP抑制剂MK 886共有的特征。尽管AA释放、环加氧酶和相关LO未受影响，但BRP-187也有效抑制微粒体前列腺素E-2合酶-1（IC 50 = 0.2 μ M），这是另一个MAPEG成员。在体内，BRP-187（10 mg/kg）在酵母多糖诱导的小鼠腹膜炎中表现出显著的有效性，抑制腹膜渗出液中的LT水平以及血管通透性和中性粒细胞浸润。总之，BRP-187在体外和体内有效地抑制LT生物合成，这似乎是通过阻止5-LO/FLAP复合物组装引起的，并保证进一步的临床前评价。(C)2016 Elsevier Inc. All rights reserved.
The pro-inflammatory leukotrienes (LTs) are formed from arachidonic acid (AA) in activated leukocytes, where 5-lipoxygenase (5-LO) translocates to the nuclear envelope to assemble a functional complex with the integral nuclear membrane protein 5-LO-activating protein (FLAP). FLAP, a MAPEG family member, facilitates AA transfer to 5-LO for efficient conversion, and LT biosynthesis critically depends on FLAP. Here we show that the novel LT biosynthesis inhibitor BRP-187 prevents the 5-LO/FLAP interaction at the nuclear envelope of human leukocytes without blocking 5-LO nuclear redistribution. BRP-187 inhibited 5-LO product formation in human monocytes and polymorphonuclear leukocytes stimulated by lipopolysaccharide plus N-formyl-methionyl-leucyl-phenylalanine (IC50 = 7-10 nM), and upon activation by ionophore A23187 (IC50 = 10-60 nM). Excess of exogenous AA markedly impaired the potency of BRP-187. Direct 5-LO inhibition in cell-free assays was evident only at >35-fold higher concentrations, which was reversible and not improved under reducing conditions. BRP-187 prevented A23187-induced 5-LO/FLAP complex assembly in leukocytes but failed to block 5-LO nuclear translocation, features that were shared with the FLAP inhibitor MK886. Whereas AA release, cyclooxygenases and related LOs were unaffected, BRP-187 also potently inhibited microsomal prostaglandin E-2 synthase-1 (IC50 = 0.2 mu M), another MAPEG member. In vivo, BRP-187 (10 mg/kg) exhibited significant effectiveness in zymosan-induced murine peritonitis, suppressing LT levels in peritoneal exudates as well as vascular permeability and neutrophil infiltration. Together, BRP-187 potently inhibits LT biosynthesis in vitro and in vivo, which seemingly is caused by preventing the 5-LO/FLAP complex assembly and warrants further preclinical evaluation. (C) 2016 Elsevier Inc. All rights reserved.