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.

项目摘要/摘要 细菌感染是死亡，健康损害和残疾的主要原因。不幸的是，我们是 目前目睹了多次抗感染的增加，而新的发展也有所减少 抗菌剂。因此，治疗成本正在增加，越来越多的患者是 屈服于这些感染。此外，难以治疗甚至不可治疗的细菌的增加 妥协医疗程序，例如治疗癌症和其他慢性疾病，手术，器官 移植，牙科工作和对早产婴儿的护理。复杂问题，因为SARS-COV-2 与多药耐药细菌的共同感染已经记录在案，Covid-19大流行可能 通过增加患者暴露于抗菌剂的情况来加快抗生素耐药性的升高。解决方案 抗生素耐药性问题可能是新类抗菌药物的持续发展。然而， 这条路线缓慢而昂贵，需要通过其他策略来完成。此提案回应 这种需求并集中于搜索策略，以延长当前可用药物的使用寿命。 氨基糖苷6'-N-乙酰转移酶类型IB [AAC（6'）-IB]负责大多数抗性的情况 革兰氏阴性病原体中的amikacin（AK）和其他氨基糖苷。 AAC（6'）-IB的传播 这些病原体中的基因侵蚀了这些抗生素的效率，这是 反对严重感染的武术。这项研究的长期目标是开发化合物 这降低了AK对敏感性水平的抗性，可以用作治疗AK耐药感染的调节器。 该项目的特定目的1提案，以优化与细胞穿透辣椒（CPP）结构的结构 寡核苷酸类似物，称为外部指南序列（EGSS），结合了整个区域 AAC（6'） - Ib mRNA并形成RNase P的底物，该酶P裂解了mRNA阻止翻译的mRNA。这 计划的实验包括设计抗蛋白酶的CPP，以最大化内在化和测试 由脱氧核糖核苷酸和最新一代的桥核酸组成的嵌合寡聚物。 特定目标2将使用组合库鉴定AAC（6'） - IB的小分子抑制剂并优化 它们通过结构活动关系分析。该特定目的还建议设计水溶性 与锌离子复合物中的离子载是AK酶失活的强抑制剂。具体目标 3包括测试AK的效果以及与在 先前针对大肠杆菌，克雷伯氏菌和鲍曼尼杆菌模型的特定目标 使用三维棋盘测定法，时间杀害测定和感染治疗的菌株 Galleria Mellonella感染模型。然后，最有希望的组合将在大约100次上进行测试 基因型定义明确的K.肺炎，A。Baumannii，铜绿假单胞菌和肠杆菌cloacae 临床分离株。

项目成果

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

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.

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

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

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