McAuley S, Huynh A, Czarny T L, Brown E D, Nodwell J R
Biochemistry , University of Toronto , Toronto , ON , Canada . Email:
Biochemistry and Biomedical Sciences , McMaster University , Hamilton , ON , Canada.
Medchemcomm. 2018 Feb 15;9(3):554-561. doi: 10.1039/c8md00009c. eCollection 2018 Mar 1.
Small molecule disruption of the bacterial membrane is both a challenge and interest for drug development. While some avoid membrane activity due to toxicity issues, others are interested in leveraging the effects for new treatments. Existing assays are available for measuring disruption of membrane potential or membrane permeability, two key characteristics of the bacterial membrane, however they are limited in their ability to distinguish between these properties. Here, we demonstrate a high throughput assay for detection and characterization of membrane active compounds. The assay distinguishes the effect of small molecules on either the membrane potential or membrane permeability using the fluorescent dyes TO-PRO-3 iodide and DiOC(3) without the need for secondary assays. We then applied this assay to a library of 3520 synthetic molecules previously shown to inhibit growth of in order to determine the frequency of membrane activity within such a biologically active library. From the library, we found 249 compounds that demonstrated significant membrane activity, suggesting that synthetic libraries of this kind do not contain a plurality of membrane active molecules.
小分子破坏细菌膜对药物开发来说既是一项挑战，也是一个研究热点。尽管有些人因毒性问题而避免使用膜活性药物，但其他人则对利用其效果开发新疗法感兴趣。现有的检测方法可用于测量膜电位或膜通透性的破坏情况，这是细菌膜的两个关键特性，然而它们区分这些特性的能力有限。在此，我们展示了一种用于检测和表征膜活性化合物的高通量检测方法。该检测方法使用荧光染料碘化丙啶和DiOC(3)来区分小分子对膜电位或膜通透性的影响，无需进行二次检测。然后，我们将此检测方法应用于一个先前已证明能抑制生长的3520个合成分子的文库，以确定此类生物活性文库中膜活性的频率。从该文库中，我们发现了249种具有显著膜活性的化合物，这表明这类合成文库并不包含大量的膜活性分子。
