title: Active fragments-guided drug discovery and design of selective tropane alkaloids using ultra-high performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry coupled with virtual calculation and biological evaluation.
abstract: Tropane alkaloids (TAs), rich in the plant of Physochlaina infundibularis Kuang, which is named Huashanshen (HSS) in China, showed good effects on types of spasms. However, no data were collected to explore the relationship between the specificity for muscarinic receptor subtypes and the structures of these TAs. To address this issue, an extracted ion chromatogram (EIC) strategy using ultra-high performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UPLC-Q/TOF-MS) based on the fragmentation behavior of the TA standards was established to rapidly capture the varied TAs from HSS. Based on the provided structural information of diagnostic ions or neutral loss, 29 TAs were efficiently profiled, especially some trace ingredients. In additional, via virtual validation combined with molecular dynamic simulation, approximately a dozen alkaloids were found with high selectivity for muscarinic receptors. In additional, N-acetyl convolicine was chosen for selectivity evaluation of M2or M3receptors through the use of a dual-luciferase reporter assay system at the cellular level and an ACh-induced constricted strip test in vitro. After summarizing the active fragments and the structure-activity relationship (SAR) information, a new modified TA that takes advantage of both the high affinity and high selectivity for M3receptors was proposed and evaluated successfully. This study provided an effective approach for the discovery and design of natural products based on highly selective drugs by UPLC-Q/TOF-MS coupled with virtual calculation and biological evaluation. Graphical Abstract Active fragments-guided strategy for selective inhibitors from HSS.
authors: State Key Laboratory of Medicinal Chemical Biology and College of Pharmacy, Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin, 300350, China.
authors: Mengge
authors: M
authors: Zhou
authors: State Key Laboratory of Medicinal Chemical Biology and College of Pharmacy, Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin, 300350, China.
authors: Xiaoyao
authors: X
authors: Ma
authors: State Key Laboratory of Medicinal Chemical Biology and College of Pharmacy, Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin, 300350, China.
authors: Jixue
authors: J
authors: Sun
authors: State Key Laboratory of Medicinal Chemical Biology and College of Pharmacy, Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin, 300350, China.
authors: Guoyu
authors: G
authors: Ding
authors: State Key Laboratory of Medicinal Chemical Biology and College of Pharmacy, Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin, 300350, China.
authors: Qingxin
authors: Q
authors: Cui
authors: The Tianjin Engineering Center of injection package, China Otsuka Pharmaceutical Company Limited, Tianjin, 300382, China.
authors: Yan
authors: Y
authors: Miao
authors: State Key Laboratory of Medicinal Chemical Biology and College of Pharmacy, Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin, 300350, China.
authors: Yuanyuan
authors: Y
authors: Hou
authors: State Key Laboratory of Medicinal Chemical Biology and College of Pharmacy, Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin, 300350, China.
authors: Min
authors: M
authors: Jiang
authors: State Key Laboratory of Medicinal Chemical Biology and College of Pharmacy, Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin, 300350, China. gangbai@nankai.edu.cn.
authors: Gang
authors: G
authors: Bai
mesh_terms: D000109:Acetylcholine / Q000494:pharmacology; D000470:Alkaloids / Q000737:chemistry*; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D015195:Drug Design; D055808:Drug Discovery; D006168:Guinea Pigs; D056004:Molecular Dynamics Simulation; D009130:Muscle, Smooth / Q000187:drug effects; D012015:Reference Standards; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014326:Tropanes / Q000737:chemistry*
chemical_list: D000470:Alkaloids; D014326:Tropanes; D000109:Acetylcholine
keywords: Muscarinic receptor antagonists; Receptor selectivity; Tropane alkaloids; UPLC-Q/TOF-MS; Virtual screening