Dealing with antibiotic resistance: antisense technology

应对抗生素耐药性：反义技术

• 批准号: 10514492
• 负责人: MARCELO E TOLMASKY
• 金额: $ 42.6万
• 依托单位: CALIFORNIA STATE UNIVERSITY FULLERTON
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-06-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10514492
• 关键词: 2019-nCoV; 3-Dimensional; Acetylation; Acetyltransferase; Acinetobacter baumannii; Adjuvant; Amikacin; Aminoglycoside resistance; Aminoglycosides; Antibiotic Resistance; Antibiotics; Antisense Technology; Bacteria; Bacterial Antibiotic Resistance; Bacterial Infections; Binding; Biological Assay; Biological Models; COVID-19 pandemic; Caring; Cause of Death; Cells; Cellular Structures; Centers for Disease Control and Prevention (U.S.); Characteristics; Chemicals; Chronic Disease; Clinical; Combined Modality Therapy; Communicable Diseases; Complement; Complex; Dental; Deoxyribonucleotides; Development; Drug Administration Routes; Drug resistance; Effectiveness; Enterobacter cloacae; Enzyme Kinetics; Enzymes; Escherichia coli; Exposure to; Formulation; Funding; Generations; Genes; Genotype; Goals; Gram-Negative Aerobic Bacteria; Growth Inhibitors; Health; Health Care Costs; Impairment; In Vitro; Infection; Infection prevention; Influenza A Virus, H1N1 Subtype; Invertebrates; Ionophores; Ions; Klebsiella pneumoniae; Libraries; Life; Mediating; Medical; Messenger RNA; Modeling; Multi-Drug Resistance; Multiple Bacterial Drug Resistance; Nucleic Acids; Oligonucleotides; Operative Surgical Procedures; Oral; Organ Transplantation; Patients; Peptide Hydrolases; Peptides; Persons; Pharmaceutical Preparations; Pharmacology; Positioning Attribute; Predisposition; Premature Infant; Procedures; Pseudomonas aeruginosa; RNase P; Reaction; Recovery; Research; Resistance; Route; Scanning; Structure; Structure-Activity Relationship; Technology; Testing; Time; Translations; Treatment Cost; Treatment Efficacy; Treatment Failure; Water; Work; World Health Organization; Zinc; analog; antimicrobial; bacterial resistance; base; cancer therapy; co-infection; combinatorial; cost; design; disability; divalent metal; experimental study; genetic inhibitor; improved; inhibitor; minimal inhibitory concentration; novel; novel therapeutics; nuclease; nucleic acid analog; pathogen; preservation; prevent; public health emergency; resistant strain; scaffold; screening; small molecule inhibitor; synergism; tool; water solubility; water testing

Project Summary/Abstract Bacterial infections are a leading cause of death, compromised health, and disability. Unfortunately, we are currently witnessing an increase in multiresistant infections and a decrease in the development of new antimicrobials. Consequently, the treatment costs are increasing, and a growing number of patients are succumbing to these infections. Furthermore, the increase in hard-to-treat or even untreatable bacteria also compromises medical procedures such as treatment of cancer and other chronic diseases, surgery, organ transplants, dental work, and care for premature infants. Compounding the problem, since SARS-CoV-2 coinfection with multidrug resistant bacteria has already been documented, the COVID-19 pandemic could accelerate the rise in antibiotic resistance by increasing patient exposure to antimicrobials. A solution to the antibiotic resistance problem could be the continuous development of new classes of antimicrobials. However, this route is slow and costly and needs to be complemented with other strategies. This proposal responds to this need and concentrates on searching strategies to extend the useful life of currently available drugs. The aminoglycoside 6′-N-acetyltransferase type Ib [AAC(6′)-Ib] is responsible for most cases of resistance to amikacin (Ak) and other aminoglycosides in Gram-negative pathogens. The dissemination of the aac(6′)-Ib gene among these pathogens erodes the efficacy of these antibiotics, which are an important component of the armamentarium against severe infections. The long-term goal of this research is to develop compounds that reduce Ak resistance to susceptibility levels and can be used as adjuvants to treat Ak-resistant infections. Specific aim 1 of this project proposes to optimize the structure of cell-penetrating peptides (CPP) bound to oligonucleotide analogs, known as external guide sequences (EGSs), that bind a complementary region of the aac(6′)-Ib mRNA and form a substrate for RNase P, which cleaves the mRNA preventing translation. The planned experiments consist of designing protease-resistant CPPs that maximize internalization and testing chimeric oligomers composed of deoxyribonucleotides and the newest generation of bridge nucleic acids. Specific Aim 2 will identify small molecule inhibitors of the AAC(6′)-Ib using combinatorial libraries and optimize them by structure-activity relationship analysis. This Specific aim also proposes to design water-soluble ionophores that in complex with zinc ions are strong inhibitors of the enzymatic inactivation of Ak. Specific aim 3 consists of testing the effect of Ak in association with combinations of the different compounds identified in the previous specific aims on Escherichia coli, Klebsiella pneumoniae, and Acinetobacter baumannii model strains using three-dimensional checkerboard assays, time-kill assays, and treatment of infections in the Galleria mellonella infection model. The most promising combinations will then be tested on about 100 genotypically well-defined K. pneumoniae, A. baumannii, Pseudomonas aeruginosa, and Enterobacter cloacae clinical isolates.

项目总结/摘要 细菌感染是导致死亡、健康受损和残疾的主要原因。不幸的是， 目前，多重耐药感染的增加和新的耐药病毒的发展减少， 抗菌剂因此，治疗费用正在增加，并且越来越多的患者需要治疗。 死于这些感染此外，难以治疗甚至无法治疗的细菌的增加也 危及医疗程序，例如癌症和其他慢性疾病的治疗、外科手术、器官移植、 器官移植、牙科治疗和早产儿护理。使问题更加复杂的是，自从SARS-CoV-2 多重耐药细菌的合并感染已经有记录，COVID-19大流行可能 通过增加患者对抗菌药物的暴露，加速抗生素耐药性的上升。的解决方案 抗生素耐药性问题可能是新类别抗菌剂的持续发展。然而，在这方面， 这条道路缓慢而昂贵，需要辅之以其他战略。本提案响应 这一需要，并集中在寻找策略，以延长目前可用的药物的使用寿命。的 氨基糖苷类6′-N-乙酰转移酶Ib型[AAC（6′）-Ib]是大多数耐药病例的原因。 丁胺卡那霉素（Ak）和其他氨基糖苷类药物在革兰氏阴性病原体中的作用。AAC（6′）-Ib的传播 这些病原体中的基因侵蚀了这些抗生素的功效，这些抗生素是 预防严重感染的医疗设备这项研究的长期目标是开发化合物 其降低了对敏感性水平的Ak抗性，并可用作治疗Ak抗性感染的佐剂。 本项目的具体目标1提出优化结合到细胞的细胞穿透肽（CPP）的结构。 寡核苷酸类似物，称为外部指导序列（EGS），其结合寡核苷酸的互补区域。 aac（6′）-Ib mRNA，并形成RNase P的底物，RNase P切割mRNA，阻止翻译。的 计划的实验包括设计抗蛋白酶的CPP，使其最大限度地内化和测试 由脱氧核糖核苷酸和最新一代的桥核酸组成的嵌合寡聚体。 具体目标2将使用组合文库鉴定AAC（6′）-Ib的小分子抑制剂，并优化 通过构效关系分析，该具体目标还建议设计水溶性 与锌离子络合的离子载体是Ak酶失活的强抑制剂。具体目标 3中鉴定的不同化合物的组合的组合的Ak的效果进行测试。 先前针对大肠埃希菌、肺炎克雷伯菌和鲍曼不动杆菌模型的特定目标 使用三维棋盘测定法、时间杀灭测定法和治疗中的感染， 大蜡螟感染模型。最有希望的组合将在大约100个 基因型明确的K.肺炎杆菌A.鲍曼不动杆菌、铜绿假单胞菌和阴沟肠杆菌 临床分离株。

项目成果

Aminoglycoside modifying enzymes.

• DOI: 10.1016/j.drup.2010.08.003
• 发表时间: 2010-12
• 期刊: DRUG RESISTANCE UPDATES
• 影响因子: 24.3
• 作者: Ramirez, Maria S.;Tolmasky, Marcelo E.
• 通讯作者: Tolmasky, Marcelo E.

Inhibition of cell division induced by external guide sequences (EGS Technology) targeting ftsZ.

• DOI: 10.1371/journal.pone.0047690
• 发表时间: 2012
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Sala CD;Soler-Bistué AJ;Korprapun L;Zorreguieta A;Tolmasky ME
• 通讯作者: Tolmasky ME

Role of Xer site-specific recombination in the genesis of pJHCMW1: an evolutionary hypothesis.

Xer 位点特异性重组在 pJHCMW1 发生中的作用：进化假设。

• DOI: 10.1016/j.jgar.2023.07.017
• 发表时间: 2023
• 期刊: Journal of global antimicrobial resistance
• 影响因子: 4.6
• 作者: Traglia,German;Ramirez,MariaSoledad;Tolmasky,MarceloE
• 通讯作者: Tolmasky,MarceloE

Rise and dissemination of aminoglycoside resistance: the aac(6')-Ib paradigm.

• DOI: 10.3389/fmicb.2013.00121
• 发表时间: 2013
• 期刊: Frontiers in microbiology
• 影响因子: 5.2
• 作者: Ramirez MS;Nikolaidis N;Tolmasky ME
• 通讯作者: Tolmasky ME

Whole-Genome Comparative Analysis of Two Carbapenem-Resistant ST-258 Klebsiella pneumoniae Strains Isolated during a North-Eastern Ohio Outbreak: Differences within the High Heterogeneity Zones.

• DOI: 10.1093/gbe/evw135
• 发表时间: 2016-07-03
• 期刊: Genome biology and evolution
• 影响因子: 3.3
• 作者: Ramirez MS;Xie G;Traglia GM;Johnson SL;Davenport KW;van Duin D;Ramazani A;Perez F;Jacobs MR;Sherratt DJ;Bonomo RA;Chain PS;Tolmasky ME
• 通讯作者: Tolmasky ME