Novel assay to monitor Tacrolimus levels at the point of care

摘要/摘要 2016年，美国进行了超过3.3万例器官移植，比过去增加了20% 五年了。器官移植需要终生免疫抑制，以防止移植排斥反应。 他克莫司是一种大环内酯类抗生素，是最有效的免疫抑制剂之一，占固体器官的90%。 移植受者接受它作为其维持免疫抑制的一部分。他克莫司有一个狭窄的 治疗窗口，亚治疗水平使患者面临排斥风险和超治疗水平 导致中毒。除了显著的药代动力学变异性外，据估计，25%的患者 不遵守他们的免疫抑制方案，显著增加了移植排斥和失败率 并增加了美国医疗体系的财政负担。因为移植受者需要终生 对于免疫抑制，任何有助于改进剂量和快速检测不粘连的方法都是 有很大的临床应用价值。 由于他克莫司的水平不能在护理点确定，因此检测非治疗性水平 门诊环境是具有挑战性的。患者必须在常规检查前参加单独的实验室预约 后续访问或他们在访问当天抽出的水平。在前者中，额外的负担加在了 患者需要参加额外的预约，而后者排除了对患者之前的水平进行分析 预约。延迟用药管理或发现患者不依从的情况会使患者处于 移植物排斥反应风险(亚治疗水平)或药物相关毒性(超治疗水平)。至 解决这一技术障碍，AffinEnergy计划开发一种护理点横向流动分析，将 实现对他克莫司水平的频繁、准确和负担得起的监测。我们已经生成了一个 专有捕获试剂，并确定了几种与纳米分子亲和力结合的候选检测试剂 给他克莫司。在第一阶段结束时，我们将有一个原型侧向流动分析，其建立的限度为 量化。在第二阶段，我们将扩大我们的分析的生产，优化性能特征，建立 保存条件，确定稳定性，并使用移植受者血液样本评估我们的分析方法。 该项目的成功完成将改善接受他克莫司治疗的患者的长期护理 用于免疫抑制，减轻患者和提供者的负担。

SUMMARY/ABSTRACT In 2016, over 33,000 organ transplants were performed in the United States, an increase of 20% over the past 5 years. Organ transplantation requires lifelong immunosuppression to prevent rejection of the transplant. Tacrolimus, a macrolide antibiotic, is one of the most effective immunosuppressants, with >90% of solid organ transplant recipients receiving it as part of their maintenance immunosuppression. Tacrolimus has a narrow therapeutic window, with sub-therapeutic levels putting patients at risk for rejection and supra-therapeutic levels leading to toxicity. In addition to significant pharmacokinetic variability, it is also estimated that >25% of patients are non-adherent to their immunosuppressant regimen, significantly increasing graft rejection and failure rates and increasing the financial burden on the US healthcare system. Because transplant recipients require a lifetime of immunosuppression, any methods that facilitate improved dosing and rapid detection of non-adherence are of great clinical utility. Because tacrolimus levels cannot be ascertained at the point of care, detection of non-therapeutic levels in the outpatient setting is challenging. Patients must either attend a separate lab appointment ahead of their routine follow-up visits or they have levels drawn on the day of their visit. In the former, an extra burden is placed on the patient to attend an additional appointment while the latter precludes analysis of levels prior to the patient’s appointment. Delays in medication management or identification of patient non-compliance place the patient at risk for graft rejection (sub-therapeutic levels) or drug-associated toxicities (supra-therapeutic levels). To address this technical hurdle, Affinergy plans to develop a point-of-care lateral flow assay that will enable frequent, accurate and affordable monitoring of tacrolimus levels. We have already generated a proprietary capture reagent and identified several detection reagent candidates that bind with nanomolar affinity to tacrolimus. At the conclusion of Phase I, we will have a prototype lateral flow assay with established limits of quantitation. In Phase II, we will scale up production of our assay, optimize performance characteristics, establish storage conditions, determine stability and evaluate our assay using transplant recipient blood specimens. Successful completion of this project will lead to improved long-term maintenance of patients receiving tacrolimus for immunosuppression and ease the burden on both patients and providers.