Identification of chemical compounds to inhibit the caseinolytic protease ClpXP complex and evaluate their biological activity

抑制酪蛋白分解酶 ClpXP 复合物的化合物的鉴定并评估其生物活性

• 批准号: 282324388
• 负责人: Professor Dr. Stephan A. Sieber
• 金额: --
• 依托单位: Lehrstuhl für Organische Chemie II
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2015
• 资助国家: 德国
• 起止时间: 2014-12-31 至 2018-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/282324388?language=en
• 关键词: Identification; chemical; compounds; inhibit; caseinolytic

In the search for new drugs against multiresistant pathogens we utilize a strategy which contrary to classical antibiotics does not kill bacteria but disarms their pathogenicity (virulence). The advantage of the anti-virulence approach is a lack of selective pressure leading to reduced resistance development and longer lasting drugs.Important regulators of bacterial virulence are the caseinolytic protease ClpP and is associated chaperone ClpX. While we were able to specifically inhibit ClpP and correspondingly virulence in S. aureus with beta-Lactones, their limited stability represents a major challenge for pharmacological application. This research proposal aims to identify novel ClpP and ClpX based virulence inhibitors with suitable pharmacological properties for in vivo application. In initial experiments we already identified reversible ClpP inhibitors via a high-throughput-screen (HTS). Their binding mode into ClpP will be investigated in detail by mutational studies. The outcome of these experiments will be utilized for the synthesis of optimized derivatives which will be tested for improved enzyme inhibition and anti-virulence properties. As ClpX represents an additional important target for an anti-virulence approach we will perform an HTS at the FMP in Berlin to identify the first ClpP/ClpX specific inhibitors. All screening hits will be again optimized and the most promising compounds further modified with an alkyne handle as well as with an aziridine photolinker in order to analyze their target selectivity. Finally, these molecules will be tested for their pharmacological properties and in case of suitable parameters tested for efficacy in a mouse abscess model.

在寻找针对多发性病原体的新药时，我们采用了一种与经典抗生素相反的策略不会杀死细菌，而是消除其致病性（毒力）。抗病毒方法的优点是缺乏选择性的压力导致耐药性发育降低和更长的药物。最重要的细菌毒力调节剂是酪蛋白溶质蛋白酶CLPP，并且是相关的伴侣CLPX。尽管我们能够特异性地抑制具有β-内酯的金黄色葡萄球菌中的CLPP和相应的毒力，但它们的有限稳定性代表了药理学应用的主要挑战。该研究建议旨在鉴定具有适合体内应用的合适药理特性的基于CLPP和CLPX的新型毒力抑制剂。在最初的实验中，我们已经通过高通量屏幕（HTS）确定了可逆的CLPP抑制剂。它们将通过突变研究详细研究其与CLPP的结合模式。这些实验的结果将用于合成优化的衍生物，这些衍生物将进行改进的酶抑制和抗病毒特性。由于CLPX代表了抗病毒方法的另一个重要目标，因此我们将在柏林的FMP进行HTS，以识别第一个CLPP/CLPX特异性抑制剂。将再次优化所有筛选命中率，并使用炔烃的手柄以及Aziridine光链接器进一步修改化合物，以分析其目标选择性。最后，将对这些分子的药理特性进行测试，并在适当的参数中测试了在小鼠脓肿模型中的功效。

项目成果

Reversible Inhibitors Arrest ClpP in a Defined Conformational State that Can Be Revoked by ClpX Association.

• DOI: 10.1002/anie.201507266
• 发表时间: 2015-12
• 期刊: Angewandte Chemie
• 作者: A. Pahl;Markus Lakemeyer;Marie-Theres Vielberg;Mathias W. Hackl;Jan Vomacka;V. Korotkov;M. Stein