Cardiovascular Disease & Respiratory Disorders Laboratory, Department of Cell Biology & Physiology, CSIR-Indian Institute of Chemical Biology, Kolkata, India.
Division of Organic and Biomolecular Medicine, CSIR-Indian Institute of Chemical Technology, Hyderabad, India.
Eur J Pharmacol. 2022 Sep 15;931:175187. doi: 10.1016/j.ejphar.2022.175187. Epub 2022 Aug 8.
Neutrophil elastase, a powerful physiological defence tool, may serve as drug target for diverse diseases due to its bystander effect on host cells like chronic obstructive pulmonary disease (COPD). Here, we synthesised seven novel benzoxazinone derivatives and identified that these synthetic compounds are human neutrophil elastase inhibitor that was demonstrated by enzyme substrate kinetic assay. One such compound, PD05, emerged as the most potent inhibitor with lower IC as compared to control drug sivelestat. While this inhibition is competitive based on substrate dilution assay, PD05 showed a high binding affinity for human neutrophil elastase (Kd = 1.63 nM) with faster association and dissociation rate compared to notable elastase inhibitors like ONO 6818 and AZD9668, and its interaction with human neutrophil elastase was fully reversible.Preclinical pharmacokinetic studies were performed in vitro where protein binding was found to be 72% with a high recovery rate, aqueous solubility of 194.7 μM, low permeability along with a favourable hERG. Experiments with cell line revealed that the molecule successfully prevented elastase induced rounding and retracted cell morphology and cell cytotoxicity. In mouse model PD05 is able to reduce the alveolar collapse induced by neutrophil elastase. In summary, we demonstrate the in situ, in vitro and in vivo anti-elastase potential of the newly synthesised benzoxazinone derivative PD05 and thus this could be promising candidate for further investigation as a drug for the treatment of COPD.
中性粒细胞弹性蛋白酶是一种强大的生理防御工具，由于其对慢性阻塞性肺疾病（COPD）等宿主细胞的旁观者效应，可能成为多种疾病的药物靶点。在这里，我们合成了七种新型苯并恶嗪酮衍生物，并通过酶底物动力学测定证实这些合成化合物是人类中性粒细胞弹性蛋白酶抑制剂。其中一种化合物 PD05 是最有效的抑制剂，其 IC50 比对照药物西维来司他低。虽然基于底物稀释测定，这种抑制是竞争性的，但 PD05 对人中性粒细胞弹性蛋白酶具有高结合亲和力（Kd = 1.63 nM），与 ONO 6818 和 AZD9668 等著名的弹性蛋白酶抑制剂相比，其结合和解离速率更快，其与人中性粒细胞弹性蛋白酶的相互作用是完全可逆的。在体外进行了临床前药代动力学研究，发现蛋白结合率为 72%，回收率高，水溶解度为 194.7 μM，渗透性低，同时 hERG 良好。细胞系实验表明，该分子成功阻止了弹性蛋白酶诱导的细胞变圆和回缩细胞形态和细胞毒性。在小鼠模型中，PD05 能够减少中性粒细胞弹性蛋白酶诱导的肺泡塌陷。总之，我们证明了新合成的苯并恶嗪酮衍生物 PD05 的原位、体外和体内抗弹性蛋白酶潜力，因此它可能是进一步研究作为治疗 COPD 的药物的有前途的候选药物。
