Development of a DNA Aptamer-Based Electrochemical Assay for Acetazolamide in Urine Using a Handheld Biosensor for Compliance Validation in Clinical Drug Trials

基于DNA适配体的手持式电化学检测尿液中乙酰唑胺 生物传感器在药物临床试验依从性验证中的应用 参加药物临床试验的30%至80%以上的患者不遵守试验方案， 丢弃研究药物或未使用适当剂量。这种常见的做法损害了医疗 试验期间的治疗决定，混淆试验数据的解释，影响监管审查 并可能导致药物开发的不适当终止，给制药业带来巨大损失。 在世界范围内，这一成本估计超过5600亿美元。此外，不遵守可以改变推荐的 药物的处方水平，如果批准，并有助于药物剂量相关的发病率和死亡率。现有 临床试验期间评估药物依从性的方法包括电子监测和自我报告，但 这些方法高估了协议的依从性。目标药物或标记化合物的定量测定，例如 也使用与研究药物共同配制的核黄素，但漏服剂量不能与 不遵守试验方案。药物或其标记物可以通过诸如LC/MS或MS/MS/MS/MS/MS等技术定量测定。 GC/MS，可以相对快速和准确，但可能需要样品制备和药物衍生化，或 示踪剂。此外，由于它们的尺寸和功率要求，LC/MS和GC/MS方法通常受到限制 中心实验室阻止临床试验监查员进一步询问受试者是否缺乏依从性。 临床样本的运输和冷藏产生了大量额外费用。这些挑战的解决方案 问题是便携式诊断系统，其：1）可以由各种类型的医疗专业人员使用， 在护理点（POC）的培训，2）在时间范围内提供药物或标记物浓度的准确结果 患者的内部临床试验或定期门诊访视，以及3）是评估治疗的通用平台 许多不同研究药物的方案依从性。ApolloDx-OTC生物技术团队为此聚集在一起， 拟议的项目已经证明了开发一个系统的能力，所有这些特点在一个简单的，廉价的！ 手持式诊断平台，广泛适用于通过定制开发的DNA定量任何药物靶点 适配体和密切监测试验方案的参与者的遵守。该小组的诊断系统和准备 其原型消耗品已针对包括小分子药物分析物在内的六种不同分析物进行了11次验证。在 在第一阶段，该团队将开发一种独特的DNA适体，因为它在过去已经无数次，以量化亚治疗 尿样中未代谢的标记物乙酰唑胺（ACZ）的剂量。基于适配子的手持设备的开发 尿中ACZ的测定将能够快速（10分钟内）、灵敏地测定ACZ与靶点的相关性 药物水平，从而帮助临床试验合规，生成更准确的试验数据，改善FDA的决策， 推进FDA和NIDA的使命。在第二阶段，该团队将进一步开发其ACZ适体分析， 开始寻求FDA的批准，同时扩大其研究药物和标签适体测定的剧目， 用于未来的临床试验。

Development of a DNA Aptamer-Based Electrochemical Assay for Acetazolamide in Urine Using a Handheld Biosensor for Compliance Validation in Clinical Drug Trials Thirty to over eighty percent of patients enrolled in pharmaceutical clinical trials do not comply with trial protocols by discarding the drug under investigation or not using the proper dosages. This common practice compromises the medical treatment decisions during the trials, confounds the interpretation of data from the trials, impacts the regulatory review and could result in inappropriate termination of a drug's development at huge expense to the pharmaceutical industry. Worldwide, this cost has been estimated to be in excess of $560BIL. Furthermore, non-adherence can alter recommended prescription levels of the drug, if approved, and contribute to drug dosing related morbidity and mortality. Existing methods of assessing pharmaceutical adherence during clinical trials include electronic monitoring, and self-reporting, but these methods overestimate protocol compliance. Quantitative assays for the target drug or tagging compounds, such as riboflavin, co-formulated with the drug under investigation are also used, but a missed dose cannot be differentiated from non-adherence to trial protocols. The drug or its marker can be determined quantitatively by techniques such as LC/MS or GC/MS, which can be relatively rapid and accurate, but may require sample preparation and derivatization of the drug or taggant. In addition, because of their size and power requirements, LC/MS and GC/MS methods are generally restricted to central laboratories preventing clinical trial monitors from further questioning enrollees about their lack of compliance. Significant additional costs accrue from transport and cold storage of clinical samples. The solution to these challenging problems is a portable diagnostic system that: 1) can be used by various types of medical professionals with minimal training at the point of care (POC), 2) provides accurate results for the drug or taggant concentration within the timeframe of the patient's in-house clinical trial or periodic clinic visits, and 3) is a versatile platform for evaluating treatment protocol compliance for many different investigational drugs. The ApolloDx-OTC Biotech team assembled for this proposed project has demonstrated the ability to develop a system with all of these characteristics in a simple, inexpensive! handheld diagnostic platform that is widely applicable to quantifying any drug target via custom developed DNA aptamers and closely monitoring trial protocols for enrollees' adherence. The team's diagnostic system and preparation of its prototype consumable has been validated 11 times for six different analytes including small molecule drug analytes. In Phase I, the team will develop a unique DNA aptamer, as it has numerous times in the past, to quantify a sub-therapeutic dose of the non-metabolized taggant acetazolamide (ACZ) in urine samples. Development of an aptamer-based handheld assay for ACZ in urine will enable rapid (within 10 minutes), sensitive determination of ACZ for correlation with target drug levels, thus aiding clinical trials compliance, generating more accurate trial data, improving FDA decisions, and advancing the missions of both the FDA and NIDA. In Phase II, the team will further develop its ACZ aptamer assay and begin seeking FDA approval for it while expanding its repertoire of investigational drug and taggant aptamer assays for use in future clinical trials.