High Throughput Screening: Bacterial Mechanosensitive Channels as Drug Targets

描述(申请人提供)：卫生保健行业的主要问题之一是多重耐药感染。医院中金黄色葡萄球菌等耐药细菌的继发感染，以及结核分枝杆菌的多重耐药形式，已成为全球卫生问题。这项提案的目的是分离潜在的广谱抗生素，这些抗生素可能会彻底改变我们治疗常见和多重耐药细菌感染的方式，甚至可能是生物恐怖主义的受害者。我们之前发现，微小的结构变化会导致大孔细菌机械敏感通道错开，严重损害细菌细胞的生长和生存能力。有许多特点使这些渠道成为理想的药理靶点。这些通道是：(1)在细菌生长的所有阶段都有表达，这表明激动剂甚至会攻击有长期休眠趋势的微生物，如结核分枝杆菌。(2)几乎在所有细菌物种中都有，但微生物是独有的；在任何哺乳动物中都没有发现家族成员。(3)高度保守，药物可能是广谱的。(4)最后，我们有许多工具用于该项目，包括零菌株、表达构建体，以及批准在校园使用高通量筛选(HTS)设施。总之，这项建议的目的是开发该检测方法，并筛选HTS库中的200,000种化合物，以分离出针对机械敏感通道的潜在抗生素药物的化合物。与公共卫生相关：我们建议分离出新一代抗生素，这种抗生素将是治疗多重耐药感染和其他难以治疗的感染的理想药物。分离出的抗生素还可能在紧急情况下产生影响，包括治疗生物恐怖主义行为的受害者。

DESCRIPTION (provided by applicant): One of the major problems in the health care industry is multi-drug resistant infections. Secondary infections of drug-resistant bacteria such as S. aureus in hospitals, and the multidrug resistant forms of M. tuberculosis, have become global health problems. The aim of this proposal is to isolate potential broad-spectrum antibiotic agents that could revolutionize how we treat common and multi-drug-resistant bacterial infections, and possibly even victims of bio-terrorism. We previously found that small structural changes can cause large- pore bacterial mechanosensitive channels to mis-gate, severely compromising the growth and viability of the bacterial cell. There are many features that make these channels ideal candidates for pharmacological targets. The channels are: (1) expressed in all phases of bacterial growth, suggesting that agonists would attack even microbes that have a tendency to remain dormant for long periods of time, such as M. tuberculosis. (2) found in essentially every bacterial species, but are unique to microbes; no family members are found in any mammal. (3) highly conserved, so drugs are likely be broad spectrum. (4) Finally, we have numerous tools for the project including null strains, expression constructs, and the approval for the use of the High Throughput Screening (HTS) facility on campus. In sum, the aim of this proposal is to develop the assay, and screen the 200,000 compounds in the HTS library to isolate compounds that are potential antibiotic drugs that target mechanosensitive channels. PUBLIC HEALTH RELEVANCE: We propose to isolate a new generation of antibiotics that would be ideal for treating multi-drug resistant and otherwise difficult to treat infections. The antibiotics isolated could also have implications in emergency situations including treating victims of acts of bioterrorism.