Discovery of PqsE Thioesterase Inhibitors for Pseudomonas aeruginosa Using DNA-Encoded Small Molecule Library Screening.,ACS Chemical Biology

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Discovery of PqsE Thioesterase Inhibitors for Pseudomonas aeruginosa Using DNA-Encoded Small Molecule Library Screening.
ACS Chemical Biology ( IF 3.5 ) Pub Date : 2020-01-10 , DOI: 10.1021/acschembio.9b00905
Julie S Valastyan _{1,

2} , Michael R Tota ₃ , Isabelle R Taylor ₁ , Vasiliki Stergioula ₁ , Graham A B Hone ₃ , Chari D Smith ₁ , Brad R Henke ₄ , Kenneth G Carson ₃ , Bonnie L Bassler _{1,

2}

Affiliation

Pseudomonas aeruginosa is a leading cause of hospital-acquired infections in the United States. PqsE, a thioesterase enzyme, is vital for virulence of P. aeruginosa, making PqsE an attractive target for inhibition. Neither the substrate nor the product of PqsE catalysis has been identified. A library of 550 million DNA-encoded drug-like small molecules was screened for those that bind to the purified PqsE protein. The structures of the bound molecules were identified by high throughput sequencing of the attached DNA barcodes. Putative PqsE binders with the strongest affinity features were examined for inhibition of PqsE thioesterase activity in vitro. The most potent inhibitors were resynthesized off DNA and examined for the ability to alter PqsE thermal melting and for PqsE thioesterase inhibition. Here, we report the synthesis, biological activity, mechanism of action, and early structure-activity relationships of a series of 2-(phenylcarbamoyl)benzoic acids that noncompetitively inhibit PqsE. A small set of analogs designed to probe initial structure-activity relationships showed increases in potency relative to the original hits, the best of which has an IC₅₀ = 5 μM. Compound refinement is required to assess their in vivo activities as the current compounds do not accumulate in the P. aeruginosa cytosol. Our strategy validates DNA-encoded compound library screening as a rapid and effective method to identify catalytic inhibitors of the PqsE protein, and more generally, for discovering binders to bacterial proteins revealed by genetic screening to have crucial in vivo activities but whose biological functions have not been well-defined.

中文翻译：

使用 DNA 编码的小分子文库筛选发现铜绿假单胞菌的 PqsE 硫酯酶抑制剂。

铜绿假单胞菌是美国医院获得性感染的主要原因。PqsE 是一种硫酯酶，对铜绿假单胞菌的毒力至关重要，这使得 PqsE 成为一个有吸引力的抑制目标。PqsE 催化的底物和产物均未鉴定。筛选了一个包含 5.5 亿个 DNA 编码的药物样小分子的文库，以寻找那些与纯化的 PqsE 蛋白结合的小分子。通过连接的 DNA 条形码的高通量测序鉴定结合分子的结构。在体外检测了具有最强亲和力特征的假定 PqsE 结合剂对 PqsE 硫酯酶活性的抑制作用. 最有效的抑制剂从 DNA 中重新合成，并检查了改变 PqsE 热熔解和 PqsE 硫酯酶抑制的能力。在这里，我们报告了一系列非竞争性抑制 PqsE 的 2-(苯基氨基甲酰基)苯甲酸的合成、生物活性、作用机制和早期构效关系。设计用于探测初始结构-活性关系的一小组类似物显示出相对于原始命中物的效力增加，其中最好的具有 IC ₅₀ = 5 μM。由于目前的化合物不会在铜绿假单胞菌中积累，因此需要对化合物进行改进以评估它们的体内活性胞质溶胶。我们的策略验证了 DNA 编码的化合物文库筛选是一种快速有效的方法来识别 PqsE 蛋白的催化抑制剂，更一般地说，用于发现通过基因筛选揭示的具有关键体内活性但其生物学功能没有的细菌蛋白结合物被明确定义。

更新日期：2020-01-10

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