Inhibition of the Bacterial LexA Repressor-Protease to Halt SOS Response-Mediated Resistance and Biofilm Formation
Inhibition of the Bacterial LexA Repressor-Protease to Halt SOS Response-Mediated Resistance and Biofilm Formation
批准号:
10194343
负责人:
Ana V Cheng
金额:
$3.64万
依托单位:
依托单位国家:
美国
项目类别:
财政年份:
2020
资助国家:
美国
项目状态:
已结题
起止时间:
2020-06-15 至 2022-01-31
关键词:
AddressAffinityAgricultureAmericanAntibiotic ResistanceAntibioticsAreaBacteriaBacterial InfectionsBindingBinding SitesBiologicalBiological TestingBypassCell divisionCellsCessation of lifeCleaved cellCommunitiesContractsDNADNA DamageDeveloped CountriesDigestionDimethyl SulfoxideDrug TargetingElectronsElementsEscherichia coliEvaluationExposure toGenesGenetic TranscriptionGenotoxic StressGentian VioletHealthHealth Care CostsHorizontal Gene TransferHospitalsInfectionInterdisciplinary StudyInvestigationKnowledgeLeadLibrariesMeasuresMediatingMedicalMedical DeviceMedicineMetabolismMicrobeMicrobial BiofilmsModificationMonitorMutagenesisNosocomial InfectionsOperative Surgical ProceduresOrganPatientsPeptide FragmentsPeptide HydrolasesPesticidesPharmacologic SubstanceProcessPromoter RegionsProteinsPseudomonas aeruginosaRec A RecombinasesResearchResistanceResistance developmentSOS ResponseSerial PassageShelter facilitySon of Sevenless ProteinsStainsStaphylococcus aureusStructureStructure-Activity RelationshipSurfaceTranscriptional RegulationWorkanalogbacterial resistancechronic infectionclinically relevantconfocal imagingdesignfunctional grouphigh throughput screeningimprovedin vitro testinginfectious disease treatmentinhibitor/antagonistinterdisciplinary approachmanmultidrug tolerancenovelnovel strategiespathogenpressurepreventresponsesensorsmall moleculesmall molecule inhibitortargeted treatmentweapons
中文摘要
项目摘要/摘要
抗生素的过度使用和误用给细菌带来了进化压力,迫使其改变
或绕过药物靶标或以其他方式产生抗药性,导致很大比例的
我们现有的药物和杀虫剂无效。新型抗生素暂时提供了
由于抗药性的快速发展而缓解,尽管几家制药公司
退出这一研究领域。细菌生物膜使许多疾病的治疗更加复杂
细菌感染。这些细胞集团有助于各种健康状况,并
已知会在大多数医疗设备的表面定居。此外,它们还庇护了大量的
持久细胞--不生长且对大多数抗生素耐受的“休眠”细胞。
不幸的是,大多数现有的疗法针对的是暂停在这些区域的代谢过程。
细菌的瞬时亚群。总而言之,我们正面临一场完美的抵抗风暴
容忍有可能导致数百万人死亡,并瓦解我们目前在
过程,除非我们找到根本的解决方案。
为此,我们已经确定了一个潜在的抗生素靶点-细菌SOS反应。
这种对遗传毒性应激的反应在细菌中是保守的,并与
抗性和耐受机制,包括水平基因转移、突变和细胞
组织逮捕。SOS基因的转录受到抑制蛋白LexA的抑制,该酶
切割与丝状蛋白RecA*相互作用,暴露SOS启动子区域。一个
以前的高通量筛选发现了一种有效的LexA裂解抑制剂。我们提出了一个
研究改善这种抑制物,更好地了解其作用和效果。使用初步的
以构效关系研究为指导,我们设计了一个包含22-25个类似物的文库
更深入的搜救行动,包括专门为克服潜在威胁而设计的模拟
外流挑战。此外,我们还提出了带有共价陷阱的多肽片段来模拟
LexA蛋白酶的天然底物,不可逆转地抑制其功能。使用我们最强大的
抑制剂,我们将研究LexA抑制的下游生物学效应,包括
获得性抗生素耐药性和生物被膜形成。我们还计划使用光亲和探测器来
确定抑制物在蛋白质中的结合位置和方向。独特的跨学科
这一提议的方法将阐明这些抑制剂的机制,并将为
为解决抵抗和容忍危机的新战略奠定基础。
英文摘要
Project Summary/Abstract
The overuse and misuse of antibiotics has put evolutionary pressure on bacteria to alter
or bypass the targets of drugs or otherwise develop resistance, rendering a large percentage of
our available medicines and pesticides ineffective. Novel antibiotics have afforded temporary
relief due to quick development of resistance, although several pharmaceutical companies have
withdrawn from this area of research. Bacterial biofilms further complicate treatment of many
bacterial infections. These cell conglomerates contribute to a variety of health conditions and are
known to colonize the surfaces of most medical devices. Moreover, they shelter high numbers of
persister cells— “dormant” cells which are non-growing and tolerant of most antibiotics.
Unfortunately, most existing therapies target metabolic processes which are suspended in these
transient subpopulations of bacteria. Altogether we are facing a perfect storm of resistance and
tolerance which threatens to kill millions and unravel our current approach to medicine in the
process, unless we find a radical solution.
To this end, we have identified a potential antibiotic target—the bacterial SOS response.
This response to genotoxic stress is conserved across bacteria and has been connected to
resistance and tolerance mechanisms, including horizontal gene transfer, mutagenesis, and cell
division arrest. Transcription of SOS genes is suppressed by the repressor-protease LexA, which
cleaves upon interaction with filamentous protein RecA* to expose the SOS promoter region. A
previous high throughput screen identified a potent inhibitor of LexA cleavage. We propose a
study to improve this inhibitor and better understand its action and effects. Using a preliminary
structure-activity relationship (SAR) study as a guide, we have designed a library of 22-25 analogs
for a more in-depth SAR campaign, including analogs specifically designed to overcome potential
efflux challenges. Additionally, we have proposed peptide fragments with covalent traps to mimic
the native substrate of the LexA protease and irreversibly inhibit its function. Using our most potent
inhibitors, we will investigate the downstream biological effects of LexA inhibition, including
acquired antibiotic resistance and biofilm formation. We also plan to use photoaffinity probes to
identify the inhibitor binding site and orientation within the protein. The uniquely interdisciplinary
approach of this proposal will elucidate the mechanism of these inhibitors and will lay the
groundwork for a novel strategy to address the resistance and tolerance crisis.
期刊论文(1)
专著(0)
科研奖励(0)
会议论文
DOI:
10.1016/j.bmcl.2022.128702
发表时间:
2022-06-01
期刊:
BIOORGANIC & MEDICINAL CHEMISTRY LETTERS
影响因子:
2.7
作者:
[Jaramillo, Ana Victoria Cheng, Cory, Michael B., Li, Allen, Kohli, Rahul M., Wuest, William M.]
通讯作者:
Wuest, William M.
海外基金