N30 Pharmaceuticals, LLC, Boulder, Colorado 80301, United States.
Biochemistry. 2012 Mar 13;51(10):2157-68. doi: 10.1021/bi201785u. Epub 2012 Feb 28.
N6022 is a novel, first-in-class drug with potent inhibitory activity against S-nitrosoglutathione reductase (GSNOR), an enzyme important in the metabolism of S-nitrosoglutathione (GSNO) and in the maintenance of nitric oxide (NO) homeostasis. Inhibition of GSNOR by N6022 and related compounds has shown safety and efficacy in animal models of asthma, chronic obstructive pulmonary disease, and inflammatory bowel disease [Sun, X., et al. (2011) ACS Med. Chem. Lett. 2, 402-406]. N6022 is currently in early phase clinical studies in humans. We show here that N6022 is a tight-binding, specific, and fully reversible inhibitor of GSNOR with an IC(50) of 8 nM and a K(i) of 2.5 nM. We accounted for the fact that the NAD(+)- and NADH-dependent oxidation and reduction reactions, catalyzed by GSNOR are bisubstrate in nature in our calculations. N6022 binds in the GSNO substrate binding pocket like a competitive inhibitor, although in kinetic assays it behaves with a mixed uncompetitive mode of inhibition (MOI) toward the GSNO substrate and a mixed competitive MOI toward the formaldehyde adduct, S-hydroxymethylglutathione (HMGSH). N6022 is uncompetitive with cofactors NAD(+) and NADH. The potency, specificity, and MOI of related GSNOR inhibitor compounds are also reported.
N6022 是一种新型的、首创的药物，对 S-亚硝基谷胱甘肽还原酶（GSNOR）具有强大的抑制活性，GSNOR 是 S-亚硝基谷胱甘肽（GSNO）代谢和维持一氧化氮（NO）动态平衡的重要酶。N6022 和相关化合物抑制 GSNOR 在哮喘、慢性阻塞性肺疾病和炎症性肠病的动物模型中显示出安全性和疗效[Sun, X., 等人。（2011）ACS Med. Chem. Lett. 2, 402-406]。N6022 目前正在进行人体早期临床试验。我们在这里表明，N6022 是 GSNOR 的紧密结合、特异性和完全可逆抑制剂，IC50 为 8 nM，K i 为 2.5 nM。我们考虑到 GSNOR 催化的 NAD(+) 和 NADH 依赖性氧化和还原反应本质上是双底物反应，在计算中计入了这一点。N6022 像竞争性抑制剂一样结合在 GSNO 底物结合口袋中，尽管在动力学测定中，它对 GSNO 底物表现出混合非竞争性抑制模式（MOI），对甲醛加合物 S-羟甲基谷胱甘肽（HMGSH）表现出混合竞争性 MOI。N6022 对辅因子 NAD(+) 和 NADH 是非竞争性的。还报告了相关 GSNOR 抑制剂化合物的效力、特异性和 MOI。
