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.

概括： 向病情不稳定、病情危重的住院患者提供有效且安全浓度的抗生素（或其他药物） 患者是一个令人畏惧的问题 - 例如，对于这项工作重点的所有六种抗生素，药代动力学 研究表明，预测活性浓度与实际活性浓度之间确实存在巨大差异。虽然重要 在帮助医生做出最有可能有帮助且最不可能造成伤害的决定方面已经取得了长足的进步， 这种方法仍然缺少一个重要的基石：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，从而解决长期周转问题 基于中央实验室的高频治疗药物方法的周期和成本高昂 监控。我们通过在每个步骤中构建和验证来系统地自下而上地解决问题 每个组件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），我们专注于高质量受体——经过充分表征和验证 他们自己的——然后可以被其他团体以不同的形式广泛使用和实施，只需简单地 被订购为“现成的”。我们将基于寡核苷酸的受体（适体）确定为此类试剂。 基于我们大量的初步结果，我们描绘了一个过程，该过程将导致受体组的分离， 每个候选者都以荧光格式根据金标准分析方法进行验证 患者来源的液体，在允许这些简单传感器直接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）。 我们的第二个挑战因从危重患者身上采集的样本的高度可变性而加剧， 是交叉反应性和与标准化条件的偏差。我们通过识别对来解决这些问题 每种抗生素具有正交特性的适配体：其中一个适配体将用于生物传感器模块 （目标 2）在严格控制的条件下结合常用的纳米材料（石墨烯）和 对透析流出物和 SPE 柱提取物进行了验证（此处，生物传感器无需预先处理即可使用） 纯化）对照金标准色谱方法。这对适体中的另一个适体来自不相关的 家族，将被全面验证为加标的提取模块（目标 3）的亲和成分 商业血清样本和危重患者的实际样本。 通过这种方法，我们将提供经过严格表征的标准化组件来分析 多粘菌素、氟喹诺酮类、达托霉素、利奈唑胺和 β 内酰胺，这将使我们（和其他人）能够 将组件组合成设备，可以在床边使用，也可以作为自动分析仪中的卡盒。 无论哪种情况，这些都将有助于大型学术医院之外的常规高频药物监测 设置。我们预计在此资助期结束时展示一种多模块设备的设计。