New anti-tubercular agents are urgently needed to address the emerging threat of drug resistance to human tuberculosis. Here, we have used structure-assisted methods to develop compounds that target mycobacterial membrane protein large 3 (MmpL3). MmpL3 is essential for the transport of mycolic acids, an important cell-wall component of mycobacteria. We prepared compounds that potently inhibit the growth of Mycobacterium tuberculosis (Mtb) and other mycobacteria in cell culture. The cryoelectron microscopy (cryo-EM) structure of mycobacterial MmpL3 in complex with one of these compounds (ST004) was determined using lipid nanodiscs at an overall resolution of 3.36 Å. The structure reveals the binding mode of ST004 to MmpL3, with the S4 and S5 subsites of the inhibitor-binding pocket in the proton translocation channel playing vital roles. These data are a promising starting point for the development of anti-tuberculosis drugs that target MmpL3.

迫切需要新的抗结核药物来解决新出现的人类结核病耐药性威胁。在这里，我们使用结构辅助方法来开发靶向分枝杆菌膜蛋白大 3 (MmpL3) 的化合物。MmpL3 对分枝杆菌酸的转运至关重要，分枝杆菌酸是分枝杆菌的重要细胞壁成分。我们制备了有效抑制结核分枝杆菌( Mtb ) 生长的化合物) 和细胞培养中的其他分枝杆菌。分枝杆菌 MmpL3 与其中一种化合物 (ST004) 复合的低温电子显微镜 (cryo-EM) 结构是使用脂质纳米圆盘以 3.36 Å 的总分辨率测定的。该结构揭示了 ST004 与 MmpL3 的结合模式，质子易位通道中抑制剂结合口袋的 S4 和 S5 亚位点起着至关重要的作用。这些数据是开发针对 MmpL3 的抗结核药物的有希望的起点。