Transformation of a Metal Chelate into a "Catch and Anchor" Inhibitor of Botulinum A Protease.
Transformation of a Metal Chelate into a "Catch and Anchor" Inhibitor of Botulinum A Protease.
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将金属螯合物转化为肉毒杆菌蛋白酶的“捕获和锚定”抑制剂。
DOI：
10.3390/ijms24054303
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发表时间：
2023-02-21
影响因子：
5.6
通讯作者：
Janda, Kim
中科院分区：
文献类型：
--
作者：
Lin, Lucy;Patel, Ealin N.;Nielsen, Alexander L.;Turner, Lewis D.;Tepp, William H.;Nguyen, Kong;Pellett, Sabine;Janda, Kim
靶向肉毒杆菌神经毒素轻链（LC）金属蛋白酶使用小分子金属螯合物抑制剂是一种很有前途的方法来对抗致命毒素的影响。然而，为了克服与简单可逆金属螯合物抑制剂相关的陷阱，研究替代支架/策略至关重要。与Atomwise Inc.合作，进行了计算机和体外筛选，产生了许多先导化合物，包括新型9-羟基-4H-吡啶并[1，2-a]嘧啶-4-酮（PPO）支架。从这个结构中，合成并测试了另外一系列的43种衍生物，得到了在BoNT/A LC酶测定中Ki为150 nM，在基于运动神经元细胞的测定中Ki为17 µM的先导候选物。这些数据结合结构-活性关系（SAR）分析和对接导致双功能设计策略，我们将其称为“捕获和锚”，用于BoNT/A LC的共价抑制。对从该捕获和锚活动制备的结构进行动力学评价，提供kinact/Ki值，以及观察到的抑制原理。通过其他测定法（包括FRET终点测定法、质谱法和极限酶透析）验证了共价修饰。所呈现的数据支持PPO支架作为BoNT/A LC的靶向共价抑制的新候选物。
Targeting the botulinum neurotoxin light chain (LC) metalloprotease using small-molecule metal chelate inhibitors is a promising approach to counter the effects of the lethal toxin. However, to overcome the pitfalls associated with simple reversible metal chelate inhibitors, it is crucial to investigate alternative scaffolds/strategies. In conjunction with Atomwise Inc., in silico and in vitro screenings were conducted, yielding a number of leads, including a novel 9-hydroxy-4H-pyrido [1,2-a]pyrimidin-4-one (PPO) scaffold. From this structure, an additional series of 43 derivatives were synthesized and tested, resulting in a lead candidate with a Ki of 150 nM in a BoNT/A LC enzyme assay and 17 µM in a motor neuron cell-based assay. These data combined with structure-activity relationship (SAR) analysis and docking led to a bifunctional design strategy, which we termed “catch and anchor” for the covalent inhibition of BoNT/A LC. Kinetic evaluation was conducted on structures prepared from this catch and anchor campaign, providing kinact/Ki values, and rationale for inhibition seen. Covalent modification was validated through additional assays, including an FRET endpoint assay, mass spectrometry, and exhaustive enzyme dialysis. The data presented support the PPO scaffold as a novel candidate for targeted covalent inhibition of BoNT/A LC.
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影响因子：
7.3
作者：
Silhár P;Eubanks LM;Seki H;Pellett S;Javor S;Tepp WH;Johnson EA;Janda KD
通讯作者：
Janda KD
DOI：
10.1073/pnas.0408526102
发表时间：
2005-07-12
影响因子：
11.1
作者：
Wein, LM;Liu, YF
通讯作者：
Liu, YF
影响因子：
4.9
作者：
Jereb, M;Zupan, M;Stavber, S
通讯作者：
Stavber, S
影响因子：
7.3
作者：
Irwin JJ;Duan D;Torosyan H;Doak AK;Ziebart KT;Sterling T;Tumanian G;Shoichet BK
通讯作者：
Shoichet BK
影响因子：
2.8
作者：
Cocco, Antoniangela;Albanese, Alberto
通讯作者：
Albanese, Alberto