A new strategy for hit generation: Novel in cellulo active inhibitors of CYP121A1 from Mycobacterium tuberculosis via a combined X-ray crystallographic and phenotypic screening approach (XP screen).
A new strategy for hit generation: Novel in cellulo active inhibitors of CYP121A1 from Mycobacterium tuberculosis via a combined X-ray crystallographic and phenotypic screening approach (XP screen).
复制标题
DOI：
10.1016/j.ejmech.2022.114105
复制
发表时间：
2022-02-15
影响因子：
6.7
通讯作者：
Abell C
中科院分区：
文献类型：
--
作者：
Frederickson M;Selvam IR;Evangelopoulos D;McLean KJ;Katariya MM;Tunnicliffe RB;Campbell B;Kavanagh ME;Charoensutthivarakul S;Blankley RT;Levy CW;de Carvalho LPS;Leys D;Munro AW;Coyne AG;Abell C
迫切需要治疗结核病(TB)的新药，以对抗当前抗结核药物日益增长的耐药性。在这里，描述了一种新的策略，针对有希望的结核目标产生命中，包括X射线晶体筛选和表型筛选相结合。这种组合方法(XP Screen)既提供了目标参与的验证，也提供了对纤维素活性的确定。通过对来自结核分枝杆菌(Mtb)的一种细胞色素P450酶CYP121A1的XP筛选来说明该方法的实用性，该酶被认为是有效的药物发现目标。用这种方法合成并测试了一个聚焦筛选集，该集的几个成员显示出对结核分枝杆菌H37Rv株的良好活性。观察到一种化合物对CYP121A1具有X射线打击作用，并在多种检测条件下对结核分枝杆菌H37Rv显示出更高的活性(泛测定活性)。在X射线晶体筛选期间获得的数据被用于基于结构的活动来设计有限数量的类似物(不到20个)，其中许多也显示出对结核分枝杆菌H37Rv株的泛分析活性。其中包括苯并[b][1，4]恶嗪衍生物(MIC906.25或μM)，这是一种新的热门化合物，适合作为更复杂的HIT来领导候选药物化学活动的起点。此前，结核分枝杆菌的细胞色素P121已被证明是分枝杆菌生存的关键靶点。以前采用的策略已经确定了高亲和力的抑制剂，然而这些药物对结核分枝杆菌缺乏活性。这里报告的策略使用X射线结晶学和表型筛选(XP筛选)相结合的方法来鉴定化合物。XP Screen方法鉴定出一些化合物表现出良好的亲和力(高达3.2UμM)和最低抑菌浓度(MIC)抗结核分枝杆菌(高达6.25MμM)。
There is a pressing need for new drugs against tuberculosis (TB) to combat the growing resistance to current antituberculars. Herein a novel strategy is described for hit generation against promising TB targets involving X-ray crystallographic screening in combination with phenotypic screening. This combined approach (XP Screen) affords both a validation of target engagement as well as determination of in cellulo activity. The utility of this method is illustrated by way of an XP Screen against CYP121A1, a cytochrome P450 enzyme from Mycobacterium tuberculosis (Mtb) championed as a validated drug discovery target. A focused screening set was synthesized and tested by such means, with several members of the set showing promising activity against Mtb strain H37Rv. One compound was observed as an X-ray hit against CYP121A1 and showed improved activity against Mtb strain H37Rv under multiple assay conditions (pan-assay activity). Data obtained during X-ray crystallographic screening were utilized in a structure-based campaign to design a limited number of analogues (less than twenty), many of which also showed pan-assay activity against Mtb strain H37Rv. These included the benzo[b][1,4]oxazine derivative (MIC90 6.25 μM), a novel hit compound suitable as a starting point for a more involved hit to lead candidate medicinal chemistry campaign. CYP121 from M.tuberculosis has been previously shown to be a crucial target for the survival of the mycobacteria. Strategies previously employed have identified high affinity inhibitors however these have lacked activity on M.tuberculosis. The strategy reported here uses a combination of X-ray crystallography and phenotypic screening (XP Screen) to identify compounds. The XP screen approach identified a number of compounds which show good affinity (up to 3.2 μM) and MIC against M.tuberculosis (up to 6.25 μM).
登录
查看更多内容
影响因子：
15
作者：
Knapp DM;Gillis EP;Burke MD
通讯作者：
Burke MD
影响因子：
3.4
作者：
Kavanagh, Madeline E.;Gray, Janine L.;Gilbert, Sophie H.;Coyne, Anthony G.;McLean, Kirsty J.;Davis, Holly J.;Munro, Andrew W.;Abell, Chris
通讯作者：
Abell, Chris
影响因子：
7.3
作者：
Harrison, James;Cox, Jonathan A. G.
通讯作者：
Cox, Jonathan A. G.
FEM：功能增强的地图。
DOI：
10.1107/s1399004714028132
发表时间：
2015-03
期刊：
Acta crystallographica. Section D, Biological crystallography
影响因子：
--
作者：
Afonine PV;Moriarty NW;Mustyakimov M;Sobolev OV;Terwilliger TC;Turk D;Urzhumtsev A;Adams PD
通讯作者：
Adams PD
影响因子：
7.3
作者：
Kavanagh ME;Coyne AG;McLean KJ;James GG;Levy CW;Marino LB;de Carvalho LP;Chan DS;Hudson SA;Surade S;Leys D;Munro AW;Abell C
通讯作者：
Abell C