Wu Zhili, Wu Yuchen, Niu Yanhong, Hu Qianfang, Jiang Qihua, Liao Lingbing, Qi Guorong, Lan Haoyang, Yang Xiaolan
Key Laboratory of Medical Laboratory Diagnostics of the Education Ministry, College of Laboratory Medicine, Chongqing Medical University, No. 1, Yixueyuan Road, Yuzhong Dist, Chongqing 400016, China.
Department of Respiratory and Critical Care Medicine, the first affiliated hospital of Chongqing Medical University, Chongqing 400016, China.
Bioorg Med Chem Lett. 2025 Aug 1;123:130217. doi: 10.1016/j.bmcl.2025.130217. Epub 2025 Apr 3.
Tuberculosis (TB) caused by Mycobacterium tuberculosis (Mtb) remains a global public health threat, particularly due to dormant Mtb, which necessitates prolonged drug treatment. Mycobacterium tuberculosis malate synthase (MtbMS) is a key rate-limiting enzyme in the glyoxylate shunt, essential for the survival of dormant Mtb but absent in the host. Using target-based virtual screening and biochemical approaches, we identified novel natural inhibitors of MtbMS. Molecular docking by Schrödinger and subsequent manual selection identified 11 compounds as potential inhibitors. Molecular dynamics (MD) simulations and binding-free energy analysis (MM/GBSA) demonstrated high stability and binding affinity of MtbMS with Nordihydroguaiaretic Acids (NDGA) and Meso-NDGA. NDGA and Meso-NDGA by inhibition experiment exhibited half-maximal inhibitory concentrations (IC) against MtbMS at 1.10 ± 0.01 μM and 14.29 ± 0.95 μM and by Isothermal Titration Calorimetry (ITC) showed binding constants (K) of 5.66 μM and 34.90 μM, respectively. Their minimum inhibitory concentrations (MIC) against Mtb H37Rv were 60.47 μg/mL and 30.24 μg/mL, respectively. In conclusion, natural products NDGA and Meso-NDGA are potent inhibitors of MtbMS and represent promising new scaffolds for combating dormant Mtb.
由结核分枝杆菌（Mtb）引起的结核病（TB）仍然是全球公共卫生威胁，特别是由于潜伏的Mtb，这需要长时间的药物治疗。结核分枝杆菌苹果酸合酶（MtbMS）是乙醛酸循环中的关键限速酶，对潜伏Mtb的存活至关重要，但在宿主中不存在。通过基于靶点的虚拟筛选和生化方法，我们鉴定出了MtbMS的新型天然抑制剂。使用薛定谔软件进行分子对接并随后进行人工筛选，确定了11种化合物为潜在抑制剂。分子动力学（MD）模拟和结合自由能分析（MM/GBSA）表明MtbMS与去甲二氢愈创木酸（NDGA）和内消旋-NDGA具有高稳定性和结合亲和力。通过抑制实验，NDGA和内消旋-NDGA对MtbMS的半数抑制浓度（IC）分别为1.10±0.01μM和14.29±0.95μM，通过等温滴定量热法（ITC）显示结合常数（K）分别为5.66μM和34.90μM。它们对结核分枝杆菌H37Rv的最低抑菌浓度（MIC）分别为60.47μg/mL和30.24μg/mL。总之，天然产物NDGA和内消旋-NDGA是MtbMS的有效抑制剂，是对抗潜伏Mtb的有前景的新支架。
