鉴定和生化表征抑制型 A 型肉毒神经毒素内切酶活性的类药化合物。
The identification and biochemical characterization of drug-like compounds that inhibit botulinum neurotoxin serotype A endopeptidase activity.
机构信息
Botulinum Research Center, and Department of Chemistry and Biochemistry, University of Massachusetts Dartmouth, 285 Old Westport Road, North Dartmouth, MA 02747, USA.
出版信息
Toxicon. 2010 Apr 1;55(4):818-26. doi: 10.1016/j.toxicon.2009.11.017. Epub 2009 Dec 7.
Abstract
A robust, high-throughput, two-tiered assay for screening small molecule inhibitors against botulinum neurotoxin serotype A was developed and employed to screen 16,544 compounds. Thirty-four compounds were identified as potent hits employing the first-tier assay. Subsequently, nine were confirmed as actives by our second-tier confirmatory assay. Of these, one displayed potent inhibitory efficacy, possessing an IC(50)=16 microM (+/-1.6 microM) in our in vitro assay. This inhibitor (0831-1035) is highly water-soluble, and possesses an IC(50)=47 microM (+/-7.0 microM) in our primary cell culture assay (with virtually no cytotoxicity up to 500 microM), suggesting that this inhibitor is a good candidate for further development as a therapeutic countermeasure to treat botulism resulting from botulinum neurotoxin serotype A intoxication. An enzyme kinetics study indicated that this inhibitor exhibits mixed non-competitive inhibition, with a K(I)=9 microM.
摘要
开发了一种针对 A 型肉毒神经毒素的高通量、两级筛选小分子抑制剂的方法，并用于筛选 16544 种化合物。采用第一级筛选方法，发现 34 种化合物具有较强的抑制活性。随后，通过我们的二级确证性测定法确认了其中 9 种化合物为活性化合物。其中一种在我们的体外测定中表现出很强的抑制效果，IC(50)=16 μM（+/-1.6 μM）。该抑制剂（0831-1035）具有高水溶性，在我们的原代细胞培养测定中（最高 500 μM 时几乎无细胞毒性）IC(50)=47 μM（+/-7.0 μM），这表明该抑制剂是进一步开发治疗 A 型肉毒神经毒素中毒引起的肉毒中毒的治疗对策的良好候选物。酶动力学研究表明，该抑制剂表现出混合非竞争性抑制，K(I)=9 μM。
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