Alturki Mansour S, Fuanta Ngolui Rene, Jarrard Madison A, Hobrath Judith V, Goodwin Douglas C, Rants'o Thankhoe A, Calderón Angela I
Department of Drug Discovery and Development, Harrison School of Pharmacy, 4306 Walker Building, Auburn University, Auburn, AL 36849, USA; Department of Pharmaceutical Chemistry, College of Clinical Pharmacy, Imam Abdulrahman Bin Faisal University, Dammam 34212, Saudi Arabia.
Department of Chemistry and Biochemistry, College of Sciences and Mathematics, 179 Chemistry Building, Auburn University, Auburn, AL 36849, USA.
Bioorg Med Chem Lett. 2018 Feb 15;28(4):802-808. doi: 10.1016/j.bmcl.2017.12.002. Epub 2017 Dec 5.
Single dose high-throughput screening (HTS) followed by dose-response evaluations is a common strategy for the identification of initial hits for further development. Early identification and exclusion of false positives is a cost-saving and essential step in early drug discovery. One of the mechanisms of false positive compounds is the formation of aggregates in assays. This study evaluates the mechanism(s) of inhibition of a set of 14 compounds identified previously as actives in Mycobacterium tuberculosis (Mt) cell culture screening and in vitro actives in Mt shikimate kinase (MtSK) assay. Aggregation of hit compounds was characterized using multiple experimental methods, LC-MS, HNMR, dynamic light scattering (DLS), transmission electron microscopy (TEM), and visual inspection after centrifugation for orthogonal confirmation. Our results suggest that the investigated compounds containing oxadiazole-amide and aminobenzothiazole moieties are false positive hits and non-specific inhibitors of MtSK through aggregate formation.
单剂量高通量筛选（HTS）随后进行剂量反应评估是识别用于进一步开发的初始活性化合物的常用策略。早期识别和排除假阳性是早期药物发现中节省成本且至关重要的一步。假阳性化合物的机制之一是在检测中形成聚集体。本研究评估了一组先前在结核分枝杆菌（Mt）细胞培养筛选中被鉴定为活性化合物以及在Mt莽草酸激酶（MtSK）检测中为体外活性化合物的14种化合物的抑制机制。使用多种实验方法对活性化合物的聚集进行了表征，包括液相色谱 - 质谱联用（LC-MS）、核磁共振氢谱（HNMR）、动态光散射（DLS）、透射电子显微镜（TEM）以及离心后的目视检查，以进行正交确认。我们的结果表明，所研究的含有恶二唑 - 酰胺和氨基苯并噻唑部分的化合物是假阳性活性化合物，并且通过聚集体形成对MtSK具有非特异性抑制作用。
