Salla Manohar, Butler Mark S, Pelingon Ruby, Kaeslin Geraldine, Croker Daniel E, Reid Janet C, Baek Jong Min, Bernhardt Paul V, Gillam Elizabeth M J, Cooper Matthew A, Robertson Avril A B
Institute for Molecular Bioscience, The University of Queensland , Brisbane, Queensland 4072, Australia.
School of Chemistry and Molecular Biosciences, The University of Queensland , Brisbane, Queensland 4072, Australia.
ACS Med Chem Lett. 2016 Sep 27;7(12):1034-1038. doi: 10.1021/acsmedchemlett.6b00198. eCollection 2016 Dec 8.
MCC950 is an orally bioavailable small molecule inhibitor of the NOD-like receptor pyrin domain-containing protein 3 (NLRP3) inflammasome that exhibits remarkable activity in multiple models of inflammatory disease. Incubation of MCC950 with human liver microsomes, and subsequent analysis by HPLC-MS/MS, revealed a major metabolite, where hydroxylation of MCC950 had occurred on the 1,2,3,5,6,7-hexahydro--indacene moiety. Three possible regioisomers were synthesized, and coelution using HPLC-MS/MS confirmed the structure of the metabolite. Further synthesis of individual enantiomers and coelution studies using a chiral column in HPLC-MS/MS showed the metabolite was -(+)- -((1-hydroxy-1,2,3,5,6,7-hexahydro--indacen-4-yl)carbamoyl)-4-(2-hydroxypropan-2-yl)furan-2-sulfonamide (). Incubation of MCC950 with a panel of cytochrome P450 enzymes showed P450s 2A6, 2C9, 2C18, 2C19, 2J2, and 3A4 catalyze the formation of the major metabolite , with a lower level of activity shown by P450s 1A2 and 2B6. All of the synthesized compounds were tested for inhibition of NLRP3-induced production of the pro-inflammatory cytokine IL-1β from human monocyte derived macrophages. The identified metabolite was 170-fold less potent than MCC950, while one regioisomer had nanomolar inhibitory activity. These findings also give first insight into the SAR of the hexahydroindacene moiety.
MCC950是一种口服生物可利用的小分子抑制剂，可抑制含NOD样受体吡啉结构域的蛋白3（NLRP3）炎性小体，在多种炎症性疾病模型中表现出显著活性。将MCC950与人肝微粒体孵育，随后通过HPLC-MS/MS分析，发现了一种主要代谢物，MCC950在1,2,3,5,6,7-六氢茚并[1,2-b]呋喃部分发生了羟基化。合成了三种可能的区域异构体，并使用HPLC-MS/MS进行共洗脱，证实了代谢物的结构。进一步合成单个对映体，并使用HPLC-MS/MS中的手性柱进行共洗脱研究，结果表明该代谢物为-(+)- -((1-羟基-1,2,3,5,6,7-六氢茚并[1,2-b]呋喃-4-基)氨基甲酰)-4-(2-羟基丙烷-2-基)呋喃-2-磺酰胺。将MCC950与一组细胞色素P450酶孵育，结果表明P450 2A6、2C9、2C18、2C19、2J2和3A4催化主要代谢物的形成，P450 1A2和2B6的活性较低。对所有合成化合物进行了测试，以抑制NLRP3诱导的人单核细胞衍生巨噬细胞产生促炎细胞因子IL-1β。鉴定出的代谢物的效力比MCC950低170倍，而一种区域异构体具有纳摩尔级的抑制活性。这些发现也首次深入了解了六氢茚并[1,2-b]呋喃部分的构效关系。
