A ligand-observed mass spectrometry approach integrated into the fragment based lead discovery pipeline.
A ligand-observed mass spectrometry approach integrated into the fragment based lead discovery pipeline.
复制标题
集成到基于片段的先导化合物发现流程中的配体观察质谱方法
DOI：
10.1038/srep08361
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发表时间：
2015-02-10
影响因子：
4.6
通讯作者：
Shui W
中科院分区：
文献类型：
--
作者：
Chen X;Qin S;Chen S;Li J;Li L;Wang Z;Wang Q;Lin J;Yang C;Shui W
在基于片段的导联发现(FBLD)中，为了可靠地检测和表征与目标结合的片段，需要结合多个正交技术的级联。考虑到主流筛选技术的局限性，我们提出了一种配体观察质谱学的方法来扩展工具包，增加了为困难目标建立FBLD管道的灵活性。在这项研究中，这一方法被整合到针对丙型肝炎病毒RNA聚合酶NS5B的FBLD计划中。我们的配基观察质谱分析通过两个独立的复杂鸡尾酒筛选和后续验证分析，从384个成员的片段库中发现了10个匹配。此外，这种基于MS的方法能够定量测量片段的弱结合亲和力，这与SPR分析总体上是一致的。然后将10个HIT中的5个成功地转化为片段结合络合物的X射线结构，为基于结构的抑制剂设计奠定了基础。这种基于MS的分析方法具有容量和速度高、方法开发最少、样品制备容易、材料消耗低和定量能力强等独特优势，有望成为当前片段筛选技术体系的宝贵补充。
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.
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影响因子：
--
作者：
Comess, Kenneth M.;Schurdak, Mark E.;Beutel, Bruce A.
通讯作者：
Beutel, Bruce A.
影响因子：
7.4
作者：
Clark, SM;Konermann, L
通讯作者：
Konermann, L
DOI：
10.1073/pnas.97.17.9367
发表时间：
2000-08-15
影响因子：
11.1
作者：
Erlanson, DA;Braisted, AC;Wells, JA
通讯作者：
Wells, JA
影响因子：
7.4
作者：
Chen, Xin;Li, Lixin;Shui, Wenqing
通讯作者：
Shui, Wenqing
影响因子：
7.3
作者：
Barelier, Sarah;Pons, Julien;Krimm, Isabelle
通讯作者：
Krimm, Isabelle