College of Pharmacy, Guizhou University of Traditional Chinese Medicine, Guiyang, 550025, China and Department of Chemistry, University of Namur, Rue de Bruxelles 61, 5000 Namur, Belgium.
Department of Chemistry, University of Namur, Rue de Bruxelles 61, 5000 Namur, Belgium.
Org Biomol Chem. 2021 Mar 4;19(8):1818-1826. doi: 10.1039/d1ob00138h.
An in situ screening assay for UDP-galactopyranose mutase (UGM, an essential enzyme of M. tuberculosis cell wall biosynthesis) has been developed to discover novel UGM inhibitors. The approach is based on the amide-forming reaction of an amino acid core with various cinnamic acids, followed by a direct fluorescence polarization assay to identify the best UGM binders without isolation and purification of the screened ligands. This assay allows us to perform one-pot high-throughput synthesis and screening of enzyme inhibitors in a 384-well plate format. UGM ligands were successfully identified by this technology and their inhibition levels were established from pure synthetic compounds in vitro and in a whole cell antibacterial assay. This study provides a blueprint for designing enamide structures as new UGM inhibitors and anti-mycobacterial agents.
已开发出一种用于 UDP-半乳糖吡喃糖基转移酶（UGM，分枝杆菌细胞壁生物合成的必需酶）的原位筛选测定法，以发现新型 UGM 抑制剂。该方法基于氨基酸核心与各种肉桂酸的酰胺形成反应，然后通过直接荧光偏振测定法来鉴定最佳 UGM 结合物，而无需筛选配体的分离和纯化。该测定法允许我们在 384 孔板格式中进行一锅式高通量合成和酶抑制剂筛选。已通过该技术成功鉴定出 UGM 配体，并从纯合成化合物在体外和全细胞抗菌测定中确定了它们的抑制水平。这项研究为设计烯酰胺结构作为新型 UGM 抑制剂和抗分枝杆菌剂提供了蓝图。
