T3SS needle protein inhibitors for the treatment of P. aeruginosa infection
T3SS针蛋白抑制剂用于治疗铜绿假单胞菌感染
基本信息
- 批准号:9335269
- 负责人:
- 金额:$ 29.78万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2016
- 资助国家:美国
- 起止时间:2016-08-19 至 2019-07-31
- 项目状态:已结题
- 来源:
- 关键词:ActinsAcuteAddressAnimal ModelAnimalsAnti-Bacterial AgentsAntibiotic ResistanceAntibioticsBacteremiaBacteriaBinding SitesBiological AssayCell membraneCellsCessation of lifeChemicalsClinicalCommunitiesCritical IllnessDevelopmentDrug EffluxDrug resistanceEndocarditisEnsureExhibitsFluorescenceGenetic studyGoalsGram-Negative Bacterial InfectionsGrowthHealthHumanImmuneIn VitroInfectionInjectableInnate Immune ResponseIntensive Care UnitsIntoxicationLeadLibrariesLungMammalian CellMediatingMedicalMembraneMethodsModelingMonitorNeedlesNosocomial pneumoniaOutcomePatientsPermeabilityPhagocytesPharmaceutical PreparationsPhasePneumoniaPolymersPrevalencePropertyProteinsPseudomonas aeruginosaResistanceScreening ResultSeriesStructureSystemTherapeuticTherapeutic AgentsThickToxicity TestsToxinUrinary tractVirulenceVirulence FactorsWound InfectionYersinia pestisanalogappendagebacterial resistancebasecombatdrug developmentefficacy testingefflux pumpextracellularhigh throughput screeninginhibitor/antagonistnovel strategiesnovel therapeuticspathogenpolymerizationprophylacticpublic health relevanceresistance factorsresistance mechanismresistant strainresponsescaffoldscreeningself assemblysmall moleculesmall molecule inhibitorweapons
项目摘要
DESCRIPTION (provided by applicant): The inability to treat many Gram-negative bacterial infections effectively with existing antibiotics is a major medical crisis. Pseudomonas aeruginosa is a prime example: 30% of clinical isolates from critically ill patients are resistant to three or
more drugs. The overall goal of this project is to address the critical medical need by a novel approach of identifying specific inhibitors of the type-three secretion system (T3SS) targeting the extracellular T3SS needle and developing them into novel therapeutic agents against P. aeruginosa. T3SS is the major virulence factor contributing to the establishment and dissemination of P. aeruginosa infections and is utilized by the bacterium to secrete and translocate toxin effectors into host phagocytes and weaken the host's innate immune response. The presence of a functional T3SS is significantly associated with poor clinical outcomes and death in patients and markedly reduces survival in animal infection models. T3SS inhibitors will be administered therapeutically and prophylactically in combination with anti-pseudomonal agents to inhibit the T3SS, potentiate a robust host innate immune response, and enhance the antibacterial activity of co-administered antibiotics. The strategy is to identify and optimize small molecules that interfere with the extracellular T3SS needle polymerization or stability. Such therapeutics will by-pass P. aeruginosa intrinsic resistance mechanisms caused by a poorly permeable outer membrane and efflux pumps. Preliminary studies revealed a putative binding site for the phenoxyacetamide series of T3SS inhibitors in the polymeric form of the needle protein PscF, indicating that the needle is a target of the P. aeruginosa T3SS for small molecule inhibition. The strategy of screening directly for compounds that alter the needle assembly or stability will capitalize on this newfound vulnerability and provide additional chemotypes of needle inhibitors for the drug development pipeline. In other preliminary studies, we developed methods for the purification of PscF and demonstrated a fluorescence-based assay to monitor the polymerization of a purified T3SS needle protein. In Phase I, a high-throughput screen using purified PscF will be developed, optimized and implemented to identify small molecules that inhibit needle polymerization or stability. Diverse compound libraries will be
screened, and resulting 'hits' will be confirmed in the screening assay in replicate, prioritized b potency, and selectivity by eliminating compounds that alter actin polymerization or stability or are promiscuous in multiple screens. Confirmed potent, selective 'hits' will be validated as T3SS inhibitors by determining their ability to inhibit effector secretion and translocation from P. aeruginosa, and by ensuring that they are not cytotoxic, do not disrupt mammalian cell membranes, and do not affect bacterial growth or viability in vitro. Preliminary SAR and in vitro ADME assays and will be used to prioritize analogs. In Phase II, the most promising of these T3SS inhibitors will be optimized to develop lead compounds for efficacy and toxicity testing in animal models.
描述(由申请人提供):现有抗生素无法有效治疗许多革兰氏阴性细菌感染是一个重大的医疗危机,这是一个典型的例子:来自危重患者的临床分离株有 30% 对三种或三种耐药。
该项目的总体目标是通过一种新方法来解决关键的医疗需求,该方法识别针对细胞外 T3SS 针的特定三型分泌系统 (T3SS) 抑制剂,并将其开发为针对铜绿假单胞菌的新型治疗剂。 T3SS 是导致铜绿假单胞菌感染建立和传播的主要毒力因子,细菌利用 T3SS 分泌毒素效应物并将其转移到宿主吞噬细胞中并削弱宿主的先天免疫反应与患者的不良临床结果和死亡显着相关,并且在动物感染模型中,T3SS抑制剂将与抗假单胞菌药物联合用于治疗和预防。 T3SS,增强宿主先天免疫反应,并增强共同施用的抗生素的抗菌活性。该策略是识别和优化干扰细胞外 T3SS 针的小分子。此类疗法将绕过由渗透性差的外膜和外排泵引起的铜绿假单胞菌内在耐药机制。初步研究揭示了针蛋白 PscF 聚合形式中苯氧基乙酰胺系列 T3SS 抑制剂的假定结合位点。 ,表明针是铜绿假单胞菌 T3SS 小分子抑制的靶点。直接筛选改变针组装或稳定性的化合物的策略将是。利用这一新发现的漏洞,为药物开发管道提供额外的针抑制剂化学类型,在其他初步研究中,我们开发了 PscF 的纯化方法,并演示了一种基于荧光的检测方法来监测纯化的 T3SS 针蛋白的聚合。第一阶段将开发、优化和实施使用纯化 PscF 的高通量筛选,以识别抑制针聚合或稳定性的小分子。
筛选,并在重复筛选中确认所产生的“命中”,优先进行效力测定,并通过消除改变肌动蛋白聚合或稳定性或在多次筛选中混杂的化合物来验证选择性。 T3SS抑制剂通过确定其抑制铜绿假单胞菌效应子分泌和易位的能力,并确保它们没有细胞毒性,不会破坏哺乳动物细胞膜,并且不会影响细菌生长或初步 SAR 和体外 ADME 的可行性,并将用于优先进行第二阶段的模拟测定,将优化这些最有前途的 T3SS 抑制剂,以开发用于动物模型功效和毒性测试的先导化合物。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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Joan C Mecsas其他文献
Joan C Mecsas的其他文献
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{{ truncateString('Joan C Mecsas', 18)}}的其他基金
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T3SS needle protein inhibitors for the treatment of P. aeruginosa infection
T3SS针蛋白抑制剂用于治疗铜绿假单胞菌感染
- 批准号:
9046046 - 财政年份:2016
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$ 29.78万 - 项目类别:
Initiation and regulation of antibacterial innate immunity
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8764810 - 财政年份:2014
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Initiation and regulation of antibacterial innate immunity
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