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.

在寻找针对多重耐药病原体的新药时，我们采用了一种与经典抗生素相反的策略，它不会杀死细菌，但会解除其致病性（毒力）。抗毒力方法的优点是缺乏选择性压力，从而减少耐药性的产生并延长药物的持续时间。细菌毒力的重要调节剂是酪蛋白分解酶 ClpP 及其相关分子伴侣 ClpX。虽然我们能够用 β-内酯特异性抑制 ClpP 以及相应的金黄色葡萄球菌毒力，但其有限的稳定性对药理学应用构成了重大挑战。本研究计划旨在鉴定具有适合体内应用的药理学特性的新型 ClpP 和 ClpX 毒力抑制剂。在最初的实验中，我们已经通过高通量筛选 (HTS) 鉴定了可逆的 ClpP 抑制剂。它们与 ClpP 的结合模式将通过突变研究进行详细研究。这些实验的结果将用于合成优化的衍生物，并测试其改进的酶抑制和抗毒特性。由于 ClpX 代表了抗毒力方法的另一个重要靶标，我们将在柏林的 FMP 上进行 HTS，以确定第一个 ClpP/ClpX 特异性抑制剂。所有筛选的命中将再次优化，最有希望的化合物将用炔烃手柄以及氮丙啶光联剂进一步修饰，以分析其目标选择性。最后，将测试这些分子的药理学特性，并在合适的参数的情况下测试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.

可逆抑制剂将 ClpP 抑制在特定的构象状态，该构象状态可由 ClpX 协会撤销。

• DOI: 10.1002/anie.201507266
• 发表时间: 2015-12-21
• 期刊: Angewandte Chemie
• 作者: A. Pahl;Markus Lakemeyer;Marie;Mathias W. Hackl;Jan Vomacka;V. Korotkov;M. Stein
• 通讯作者: M. Stein

A Chemical Disruptor of the ClpX Chaperone Complex Attenuates the Virulence of Multidrug-Resistant Staphylococcus aureus.

ClpX 分子伴侣复合物的化学干扰剂可减弱多重耐药金黄色葡萄球菌的毒力。

• DOI: 10.1002/anie.201708454
• 发表时间: 2017-09-14
• 期刊: Angewandte Chemie
• 作者: C. Fetzer;V. Korotkov;Robert Thänert;K. M. Lee;M. Neuenschw;er;er;J. V. von Kries;E. Medina;S. Sieber
• 通讯作者: S. Sieber