title: A ligand-observed mass spectrometry approach integrated into the fragment based lead discovery pipeline.
abstract: In fragment-based lead discovery (FBLD), a cascade combining multiple orthogonal technologies is required for reliable detection and characterization of fragment binding to the target. Given the limitations of the mainstream screening techniques, we presented a ligand-observed mass spectrometry approach to expand the toolkits and increase the flexibility of building a FBLD pipeline especially for tough targets. In this study, this approach was integrated into a FBLD program targeting the HCV RNA polymerase NS5B. Our ligand-observed mass spectrometry analysis resulted in the discovery of 10 hits from a 384-member fragment library through two independent screens of complex cocktails and a follow-up validation assay. Moreover, this MS-based approach enabled quantitative measurement of weak binding affinities of fragments which was in general consistent with SPR analysis. Five out of the ten hits were then successfully translated to X-ray structures of fragment-bound complexes to lay a foundation for structure-based inhibitor design. With distinctive strengths in terms of high capacity and speed, minimal method development, easy sample preparation, low material consumption and quantitative capability, this MS-based assay is anticipated to be a valuable addition to the repertoire of current fragment screening techniques.
authors: 1] College of Life Sciences, Nankai University, Tianjin 300071, China [2] High-throughput Molecular Drug Discovery Center, Tianjin Joint Academy of Biotechnology and Medicine, Tianjin 300457, China.
authors: Xin
authors: X
authors: Chen
authors: 1] College of Life Sciences, Nankai University, Tianjin 300071, China [2] High-throughput Molecular Drug Discovery Center, Tianjin Joint Academy of Biotechnology and Medicine, Tianjin 300457, China.
authors: Shanshan
authors: S
authors: Qin
authors: 1] College of Life Sciences, Nankai University, Tianjin 300071, China [2] High-throughput Molecular Drug Discovery Center, Tianjin Joint Academy of Biotechnology and Medicine, Tianjin 300457, China.
authors: Shuai
authors: S
authors: Chen
authors: 1] College of Life Sciences, Nankai University, Tianjin 300071, China [2] State Key Laboratory of Medicinal Chemical and Department of Pharmacy, Nankai University, Tianjin 300071, China.
authors: Jinlong
authors: J
authors: Li
authors: High-throughput Molecular Drug Discovery Center, Tianjin Joint Academy of Biotechnology and Medicine, Tianjin 300457, China.
authors: Lixin
authors: L
authors: Li
authors: 1] College of Life Sciences, Nankai University, Tianjin 300071, China [2] High-throughput Molecular Drug Discovery Center, Tianjin Joint Academy of Biotechnology and Medicine, Tianjin 300457, China.
authors: Zhongling
authors: Z
authors: Wang
authors: 1] College of Life Sciences, Nankai University, Tianjin 300071, China [2] High-throughput Molecular Drug Discovery Center, Tianjin Joint Academy of Biotechnology and Medicine, Tianjin 300457, China.
authors: Quan
authors: Q
authors: Wang
authors: 1] High-throughput Molecular Drug Discovery Center, Tianjin Joint Academy of Biotechnology and Medicine, Tianjin 300457, China [2] State Key Laboratory of Medicinal Chemical and Department of Pharmacy, Nankai University, Tianjin 300071, China.
authors: Jianping
authors: J
authors: Lin
authors: 1] High-throughput Molecular Drug Discovery Center, Tianjin Joint Academy of Biotechnology and Medicine, Tianjin 300457, China [2] State Key Laboratory of Medicinal Chemical and Department of Pharmacy, Nankai University, Tianjin 300071, China.
authors: Cheng
authors: C
authors: Yang
authors: 1] College of Life Sciences, Nankai University, Tianjin 300071, China [2] Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China.
authors: Wenqing
authors: W
authors: Shui
mesh_terms: D055808:Drug Discovery / Q000379:methods*; D016174:Hepacivirus / Q000737:chemistry / Q000378:metabolism; D006526:Hepatitis C / Q000188:drug therapy / Q000378:metabolism; D013058:Mass Spectrometry / Q000379:methods*; D017361:Viral Nonstructural Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism; D012324:RNA-Dependent RNA Polymerase
chemical_list: D017361:Viral Nonstructural Proteins; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase