URINALYSIS FOR PATIENT COMPLIANCE TO CAM THERAPY

尿液分析以确保患者对 CAM 治疗的依从性

• 批准号: 7244224
• 负责人: RAMA P RAMANUJAM
• 金额: $ 23.83万
• 依托单位: SWAASTH, INC.
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-30 至 2007-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7244224
• 关键词: body fluids; clinical trials; measurement; nuclear factor kappa beta; phosphorylation; urinalysis; urine

DESCRIPTION (provided by applicant): URINALYSIS FOR PATIENT COMPLIANCE TO CAM THERAPY: In recent years, with increasing dissatisfaction with modern medicines coupled with a desire for healthy living, there has been a dramatic increase in the use of complementary and alternative medicines (CAM). However, rigorous testing of CAM methods is needed to prove efficacy before it can be accepted in regular practice. Outpatient trials can only work if patients adhere to the therapeutic protocol. Currently, assessment of adherence is done by patient reports, pill counts and with electronic monitoring devices. The financial burden of non-adherence is estimated to cost $100 billion each year in the United States. Our goal in this application is to develop a technique to monitor patient adherence to CAM techniques using urinalysis. Towards this, we propose to develop a rapid, real-time assay to determine the effectiveness of constituents of biologically active CAM agents in body fluids. The assay is based on an in vitro bioluminescence imaging system. Ultimately, we plan to develop a non-invasive urinalysis kit. A critical cellular factor that is involved in controlling many normal cellular and organismal processes, including immune and inflammatory responses, cellular growth, and apoptosis is the transcription factor NF-kappaB. Of greater significance is that NF-kappaB is continually active in numerous disease states and is present as a latent, inactive, I-kappaB-bound complex in the cytoplasm. I-kappaB is a complex of 3 subunits. Following stimulation by an extracellular signal, the alpha subunit of I-kappaB is targeted for phosphorylation followed by ubiquitination and proteosomal degradation. Phosphorylation of I-kappaBalpha releases it from the NF-kappaB complex and the unmasked NF-kappaB complex can then enter the nucleus to activate target gene expression. CAM agents, such as curcumin, are known to inhibit I-kappaB phosphorylation, thereby inhibiting NFkappaB activation and suppressing pathophysiological processes. The primary objective of this project is to develop sensitive, non-invasive kits and assays for use in the detection of biologically active CAM agents, their constituents or metabolites in body fluids, such as blood or urine, as a way of assessing adherence to well-described clinical trial protocols. Towards this goal, we propose to validate an in vitro biological assay system using I-kappaB stabilization to determine the amount of metabolites of common CAM agents in urine (non-invasive procedure), to ultimately develop reagent kits and consumables. The Phase I effort will establish proof-of-principle for the detection and measurement of the CAM agents, such as turmeric, black raspberries and broccoli, potent anti-inflammatory agents, in the efficacy of I-kappaB degradation. I-kappaB is a critical factor involved in the inflammatory pathway and plays a vital role in NFkappaB activity. We believe that this technology will enhance and complement existing HPLC/mass spectrophotometry techniques in the accurate assessment of the use of CAM agents-before during and post clinical trials. It is anticipated that the availability of this I-kappaB degradation and measurement system will promote the development of validated CAM agents for use by consumers, patients, industry, regulatory agencies, and clinical researchers. In Addition, this GEM tool will enable both the research and commercial entities to perform molecular and functional in vitro and in vivo studies and offer exciting new opportunities in the field of complementary and alternative medicine.

描述（由申请人提供）：患者对CAM治疗的依从性的尿失禁：近年来，随着对现代药物的日益不满以及对健康生活的渴望，补充和替代药物（CAM）的使用急剧增加。然而，CAM方法需要严格的测试，以证明其有效性，然后才能在常规实践中被接受。门诊试验只有在患者遵守治疗方案的情况下才能起作用。目前，通过患者报告、药丸计数和电子监测设备来评估依从性。据估计，美国每年因不遵守《公约》而造成的财政负担高达1 000亿美元。我们在此应用中的目标是开发一种技术，以监测患者遵守CAM技术使用尿液分析。为此，我们建议开发一种快速，实时测定，以确定体液中的生物活性CAM剂的成分的有效性。该测定基于体外生物发光成像系统。最终，我们计划开发一种非侵入性尿液分析试剂盒。转录因子NF-κ B是一种参与控制许多正常细胞和生物过程的关键细胞因子，包括免疫和炎症反应、细胞生长和凋亡。更重要的是，NF-κ B在许多疾病状态中持续活跃，并在细胞质中作为潜伏的、无活性的I-κ B结合复合物存在。I-kappaB是由3个亚基组成的复合物。在细胞外信号的刺激下，I-kappaB的α亚基被靶向磷酸化，随后被泛素化和蛋白酶体降解。I-κ B α的磷酸化将其从NF-κ B复合物中释放出来，然后未掩蔽的NF-κ B复合物可以进入细胞核以激活靶基因表达。已知CAM试剂，如姜黄素，抑制I-kappaB磷酸化，从而抑制NF κ B活化并抑制病理生理过程。 该项目的主要目标是开发敏感的非侵入性试剂盒和检测方法，用于检测体液（如血液或尿液）中的生物活性CAM试剂、其成分或代谢物，作为评估对详细描述的临床试验方案的依从性的一种方式。为了实现这一目标，我们建议使用I-kappaB稳定化来验证体外生物测定系统，以确定尿液中常见CAM试剂的代谢物的量（非侵入性程序），以最终开发试剂盒和耗材。第一阶段的工作将建立检测和测量CAM试剂的原理验证，如姜黄，黑树莓和西兰花，有效的抗炎剂，在I-kappaB降解的功效。I-kappaB是参与炎症通路的关键因子，在NF-κ B活性中起着至关重要的作用。我们相信，这项技术将加强和补充现有的HPLC/质谱分光光度法在准确评估CAM剂的使用前，临床试验期间和后。预计该I-kappaB降解和测量系统的可用性将促进经验证的CAM试剂的开发，以供消费者、患者、行业、监管机构和临床研究人员使用。此外，该GEM工具将使研究和商业实体能够进行体外和体内的分子和功能研究，并在补充和替代医学领域提供令人兴奋的新机会。