Ahrens Thomas, Bergner Andreas, Sheppard David, Hafenbradl Doris
BioFocus, Allschwil, Switzerland.
J Biomol Screen. 2012 Jan;17(1):85-98. doi: 10.1177/1087057111422823. Epub 2011 Oct 11.
For many novel epigenetics targets the chemical ligand space and structural information were limited until recently and are still largely unknown for some targets. Hit-finding campaigns are therefore dependent on large and chemically diverse libraries. In the specific case of the histone methyltransferase G9a, the authors have been able to apply an efficient process of intelligent selection of compounds for primary screening, rather than screening the full diverse deck of 900 000 compounds to identify hit compounds. A number of different virtual screening methods have been applied for the compound selection, and the results have been analyzed in the context of their individual success rates. For the primary screening of 2112 compounds, a FlashPlate assay format and full-length histone H3.1 substrate were employed. Validation of hit compounds was performed using the orthogonal fluorescence lifetime technology. Rated by purity and IC(50) value, 18 compounds (0.9% of compound screening deck) were finally considered validated primary G9a hits. The hit-finding approach has led to novel chemotypes being identified, which can facilitate hit-to-lead projects. This study demonstrates the power of virtual screening technologies for novel, therapeutically relevant epigenetics protein targets.
对于许多新型表观遗传学靶点，直到最近化学配体空间和结构信息仍然有限，而且对于某些靶点在很大程度上仍然未知。因此，寻找活性化合物的研究依赖于大型且化学结构多样的文库。就组蛋白甲基转移酶G9a的具体情况而言，作者能够应用一种高效的化合物智能筛选流程用于初筛，而不是对90万种化合物的整个多样文库进行筛选来鉴定活性化合物。已应用多种不同的虚拟筛选方法进行化合物选择，并根据其各自的成功率对结果进行了分析。对于2112种化合物的初筛，采用了FlashPlate检测形式和全长组蛋白H3.1底物。使用正交荧光寿命技术对活性化合物进行验证。根据纯度和IC(50)值进行评估，最终有18种化合物（占化合物筛选文库的0.9%）被认为是经过验证的G9a初筛活性化合物。这种寻找活性化合物的方法已导致鉴定出新型化学结构类型，这有助于从活性化合物到先导化合物的研究项目。这项研究证明了虚拟筛选技术对于新型治疗相关表观遗传学蛋白质靶点的作用。
