Towards rapid measurement of antibiotics in critical care setting

在重症监护环境中快速测量抗生素

• 批准号: 10693403
• 负责人: Serge Cremers
• 金额: $ 67.62万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10693403
• 关键词: Address; Affinity; Aminoglycosides; Antibiotic Resistance; Antibiotics; Binding Proteins; Biological; Biological Assay; Biosensor; Buffers; Calibration; Chemicals; Chromatography; Clinical; Clinical Chemistry; Complex; Consensus; Coupled; Critical Care; Critical Illness; Daptomycin; Decision Making; Devices; Dialysis procedure; Dose; Drug Kinetics; Drug Monitoring; Failure; Family; Fluoroquinolones; Foundations; Frequencies; Funding; Glycopeptides; Goals; Hospitalization; Hospitals; Immunoassay; Individual; Laboratories; Linezolid; Liquid substance; Measurement; Measures; Methods; Monitor; Nature; Oligonucleotides; Organ; Patients; Pharmaceutical Preparations; Physicians; Physiological; Plasma; Polymyxins; Preparation; Process; Property; Protocols documentation; Reagent; Regimen; Renal Replacement Therapy; Reporting; Sampling; Serum; Standardization; Surface; Technology; Testing; Therapeutic; Time; Ultrafiltration; Vancomycin; Variant; Whole Blood; Work; analytical method; aptamer; bacterial resistance; beta-Lactams; candidate validation; clinical decision-making; clinical practice; compare effectiveness; cost; cost effective; cross reactivity; design; graphene; nanomaterials; neglect; new chemical entity; receptor; response; sensor; small molecule; standard of care; theories

Summary: Delivering antibiotics (or other drugs) at effective and safe concentrations to unstable, critically ill hospitalized patients is a daunting problem – for example, for all six antibiotics that are the focus of this work, pharmacokinetic studies reveal truly wide discrepancies between predicted and actual active concentrations. While important strides have been made to help physicians make decisions that are most likely to help, and least likely to harm, an important cornerstone for such approaches is still missing: There are no clinically validated devices that can rapidly, accurately and precisely measure concentrations of drugs in a manner that would address the long turn- around-times and prohibitive cost of the central-laboratory-based approach to high-frequency therapeutic drug monitoring. We approach the problem systematically from the bottom up by building and validating at each step each of the components needed to overcome such barriers. As our first challenge (and our first Aim), we focus on high-quality receptors – well characterized and validated on their own – which could then be widely used and implemented by other groups in different formats, by simply being ordered “off the shelf”. We identified oligonucleotide-based receptors (aptamers) as such reagents. Building on our vast preliminary results, we delineate a process that will lead to isolation of sets of receptors, with each candidate validated on its own in a fluorescent format against gold-standard analytical methods in patient-derived fluids, under conditions that allow these simple sensors to be applied directly in mix-and-measure formats (e.g., dialysis effluents, ultrafiltered sera, and extracts from standard SPE columns). Our second challenge, exacerbated by the highly variable nature of samples collected from critically ill patients, is cross-reactivity and deviations from standardized conditions. We address these by identifying pairs of aptamers with orthogonal properties for each antibiotic: One of the aptamers will be used in biosensor modules (Aim 2) under strictly controlled conditions in conjunction with a commonly used nanomaterial (graphene) and validated on dialysis effluents and extracts from SPE columns (here, biosensors can be used without pre- purification) against gold-standard chromatographic methods. The other aptamer from the pair, from an unrelated family, will be comprehensively validated as the affinity component of extraction modules (Aim 3) on spiked commercial samples of sera and actual samples from critically ill patients. Through this approach, we will provide rigorously characterized, standardized components for analysis of polymyxins, fluoroquinolones, daptomycin, linezolid, and beta lactams, which will enable us (and others) to combine components into devices, either to be used at the bedside, or as cartridges in automatized analyzers. In either case, these will facilitate routine high-frequency drug monitoring outside of large, academic hospital settings. We expect to demonstrate one design of multi-modular devices by the end of this funding period.

总结： 以有效和安全的浓度将抗生素（或其他药物）输送给病情不稳定的重症住院患者 例如，对于作为这项工作重点的所有六种抗生素， 研究揭示了预测的和实际的活性浓度之间确实存在很大的差异。虽然重要 在帮助医生做出最有可能帮助，最不可能伤害， 这种方法的一个重要基石仍然缺失：没有经过临床验证的设备可以 快速、准确和精确地测量药物浓度，以解决长期的问题- 以中心实验室为基础的高频治疗药物方法的周期和高昂的成本 监测.我们通过在每一步进行构建和验证，从下而上系统地解决问题 每个组成部分都需要克服这些障碍。 作为我们的第一个挑战（也是我们的第一个目标），我们专注于高质量的受体-充分表征和验证 然后可以被其他团体以不同的形式广泛使用和实施， 被命令“下架”。我们确定了基于多核苷酸的受体（适体）作为这样的试剂。 在我们大量的初步结果的基础上，我们描绘了一个过程，这将导致受体集的分离， 每种候选物都以荧光形式对照金标准分析方法进行了验证， 在允许这些简单传感器直接应用于混合和测量的条件下， 格式（例如，透析流出物、超滤血清和标准SPE柱的提取物）。 我们的第二个挑战，由于从重症患者中收集的样本的高度可变性而加剧， 是交叉反应性和标准化条件的偏差。我们通过识别成对的 对于每种抗生素具有正交特性的适体：其中一种适体将用于生物传感器模块 (Aim 2）在严格控制的条件下与常用的纳米材料（石墨烯）结合，以及 在透析流出物和SPE柱提取物上进行了验证（在此，生物传感器可以在没有预处理的情况下使用）。 纯化）相对于金标准色谱方法。另一个适体来自一个无关的 将作为加标提取模块（Aim 3）的亲和组分进行全面验证 血清的商业样品和来自重症患者的实际样品。 通过这种方法，我们将提供严格表征的标准化组件，用于分析 多粘菌素、氟喹诺酮类、达托霉素、利奈唑胺和β内酰胺类，这将使我们（和其他人） 将联合收割机组件组合到设备中，或者在床边使用，或者作为自动化分析仪中的盒。 在任何一种情况下，这些都将促进大型学术医院以外的常规高频药物监测 设置.我们希望在本资助期结束前展示一种多模块化设备的设计。