Development of a fluorescence liposomal ABCG2 Multidrug Transporter assay

荧光脂质体 ABCG2 多药物转运蛋白测定的开发

• 批准号: 8644055
• 负责人: DONALD Lee MELCHIOR
• 金额: $ 36.36万
• 依托单位: GLSYNTHESIS, INC.
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-05-07 至 2016-05-06
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8644055
• 关键词: ABCG2 gene; ATP phosphohydrolase; Adverse effects; Antineoplastic Agents; Biological Assay; Carrier Proteins; Clinical; Detergents; Development; Drug Efflux; Drug Industry; Drug Transport; Drug or chemical Tissue Distribution; Drug resistance; Excision; Fluorescence; Future; Goals; Housing; Human; In Vitro; Industry; International; Legal patent; Letters; Lipid Bilayers; Lipids; Liposomes; Malignant Neoplasms; Marketing; Materials Testing; Measurement; Mediating; Oral; P-Glycoprotein; Paper; Pharmaceutical Preparations; Predisposition; Prevalence; Production; Proteins; Protocols documentation; Pump; Reader; Reagent; Resistance; Resistance development; Robotics; Series; Services; Staging; Substrate Specificity; System; Techniques; Testing; Time; Tissues; United States; absorption; aqueous; base; chemotherapeutic agent; chemotherapy; drug candidate; drug development; drug discovery; expectation; high throughput screening; human ABCG2 protein; in vitro Assay; inhibitor/antagonist; instrumentation; multi drug transporter; multidrug transport; new technology; novel; oncology; programs; public health relevance; reconstitution; research study; scale up; sensor; tool; tumor; uptake

ABSTRACT In this project we propose to exploit the recent successful isolation and reconstitution of the important ATP- dependent drug efflux transporter, ABCG2 (BCRP, Breast Cancer Resistance Protein), and combine it with a new transport assay system, the Fluorosome platform, to characterize the interaction of ABCG2 with drugs and drug candidates. The result will provide a useful, novel, highly specific and rapid in vitro transport assay for future general use. We shall employ this assay together with ATPase studies to characterize a wide variety of ABCG2 substrates and modulators. Additionally, by means of this unique construct we shall screen the 89 anticancer drugs in the NCI Oncology Drug Set Plate series for inhibition of ABCG2-mediated transport. The ABCG2 protein is a ubiquitous high capacity drug transporter with wide substrate specificity. The "White Paper" recently issued by the International Transporter Consortium formed to propose industry and FDA standards for studies of drug:transporter interactions lists ABCG2 as second in importance to P-glycoprotein (Pgp) among those drug transporters involved in clinical absorption and disposition of drugs. Drug effects are often modified by the highly variable oral uptake of drugs and by limited tissue distribution. Tumors frequently develop resistance against treatment with multiple chemotherapeutic agents. The consequence of this is the expectation that systemic chemotherapy of nearly one-half of the one million new cancer cases annually in the United States will fail due to the resistance of tumors to drugs. A major factor in this resistance is the prevalence of energy-driven efflux transporter proteins which actively pump drugs out of their target tissues. In addition, many side effects and incompatibilities accompanying multidrug use result from the interference of one drug with another's susceptibility to active efflux. The recent successful production, isolation, and reconstitution of the ABCG2 transporter combined with new technology employing the Fluorosome platform will provide an effective in vitro assay to determine the susceptibility of drug candidates to extrusion from their target tissue by the ABCG2 transporter and to determine if compounds interact with this transporter. These studies will employ a novel reagent "Fluorosome-trans-abcg2" that is unambiguously specific for the ABCG2 transporter, applicable to a wide range of compounds, uses very small amounts of test material, and, with respect to instrumentation, requires only a standard injecting multiwell fluorescence plate reader. The assay will be amenable to moderate and high throughput screening.

抽象的 在这个项目中，我们建议利用重要的ATP-最近成功隔离和重构 依赖的药物外排转运蛋白，ABCG2（BCRP，乳腺癌耐药性蛋白），并将其与新的 运输测定系统，荧光体平台，以表征ABCG2与药物和药物的相互作用 候选人。结果将为未来提供有用的，新颖的，高度特定和快速的体外运输测定法 一般使用。我们将使用该测定法和ATPase研究来表征各种ABCG2 底物和调节剂。此外，通过这种独特的结构，我们将筛选89个抗癌 NCI肿瘤学药物集板系列中的药物抑制ABCG2介导的转运。 ABCG2蛋白是一种普遍存在的高容量的药物转运蛋白，具有广泛的底物特异性。 “白色 “国际运输商联盟最近发行的论文”，该财团旨在提出行业和FDA 药物研究标准：转运蛋白相互作用将ABCG2列为P-糖蛋白的重要性第二 （PGP）在参与药物临床吸收和处置的那些药物转运蛋白中。 药物的作用通常通过高度可变的药物吸收和有限的组织分布来改变。 肿瘤经常会产生对用多种化学治疗剂治疗的耐药性。这 结果是，预期的是，在一百万个新的一百万的全身化学疗法中 由于肿瘤对药物的抵抗力，美国每年在美国的癌症病例将失败。这是一个主要因素 抗性是能量驱动的外排转运蛋白的普遍性，该蛋白会积极从其中抽出药物 靶组织。此外，多药物使用伴随的许多副作用和不兼容。 一种药物的干扰另一个药物对积极外排的敏感性。 ABCG2转运蛋白的最近成功生产，隔离和重构与新的结合 采用荧光体平台的技术将提供有效的体外测定法，以确定 候选药物对ABCG2转运蛋白从其目标组织挤出的敏感性，并确定是否是否 化合物与该转运蛋白相互作用。这些研究将采用一种新型的试剂“荧光体 - Trans-ABCG2” 这是适用于多种化合物的ABCG2转运蛋白的明确特异性 少量的测试材料，关于仪器，仅需要标准注射多井 荧光板读取器。该测定将适合中度和高吞吐量筛选。