Combined Computational and Wet Lab Screening for Drugs Tested via OATs

对通过 OAT 测试的药物进行计算和湿实验室联合筛选

• 批准号: 7560408
• 负责人: SANJAY K NIGAM
• 金额: $ 33.8万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7560408
• 关键词: Anions; Antibiotics; Antihypertensive Agents; Antiviral Agents; Blood capillaries; Brain; Cell Line; Central Nervous System Neoplasms; Computer Simulation; Consensus; Development; Disease; Diuretics; Drug Compounding; Drug Delivery Systems; Drug Design; Drug Transport; Endothelium; Environment; Epithelial; Foundations; Funding; Goals; HIV Infections; Image; Intranasal Administration; Kidney; Knock-out; Label; Lead; Liver; Location; Mediating; Modeling; Movement; Neuraxis; Non-Steroidal Anti-Inflammatory Agents; Nose; Oats; Olfactory Epithelium; Olfactory Mucosa; Organic Anion Transporters; Pathway interactions; Pharmaceutical Preparations; Publishing; Route; Screening procedure; Side; Structure; Structure of choroid plexus; System; Tissues; United States National Institutes of Health; Validation; Wild Type Mouse; Work; Xenopus oocyte; base; brain metabolism; capillary; design; drug testing; high throughput screening; in vivo; molecular dynamics; neuropsychiatry; preference; programs; prototype; research study; small molecule libraries; tumor; virtual

DESCRIPTION (provided by applicant): This proposal ultimately seeks to develop the foundation for a virtual and wet lab high throughput screening system for drugs delivered to the central nervous system (CNS), including those useful for CNS tumors, CNS HIV infection and neuropsychiatric disorders. The goal is to identify highly specific substrates for two olfactory- expressed organic anion transporters (Oats) discovered by us, Oat6 and Oat1, thereby allowing for intranasal delivery of drugs to the CNS. Oats are considered to be the rate-limiting steps in the movement across key epithelial tissue-environment interfaces (eg. kidney, liver) of many common drugs (eg. antibiotics, antivirals, chemotherapeutics, NSAIDs, antihypertensives, diuretics). We were the first to identify the prototype, Oat1, as well as related transporters, and have recently published the Oat1 and Oat3 knockouts, which have defective organic anion transport in kidney and choroid plexus. Because of the expression of Oat6 and Oat1 at the so- called nose-brain barrier and specific substrate preferences, the pathway is attractive for a side-by-side in silico and wet lab strategy (and in vivo validation) for identification of CNS-active drugs and probes. It is hypothesized that the Oat6 and Oat1 transporters provide a unique transport mechanism for the nasal administration of drugs to the CNS, and that a side-by- side in silico (SA1) and wet lab (SA2) approach will prove synergistic and lead more rapidly to high throughput virtual and wet lab screening of a huge number of compounds to identify Oat-specific sets. These compounds may then either function as potential leads to consider for CNS-acting drugs that can be delivered nasally or, perhaps more likely, serve as a range of consensus structures for Oat6 and Oat1-mediated transport that can be considered for the design of new drugs or the derivativization of existing drugs with ineffective CNS delivery. The goal of SA1 (40-45% effort) is in silico modeling of the interaction of Oat6/Oat1 and its substrates using steered molecular dynamics in order to begin to build the basis for virtual screening. SA2 (40- 45% effort) will initially continue wet lab low-medium throughput (Xenopus oocyte transport) screening for potential Oat6 and Oat1-specific substrates and develop a high throughput assay employing stably transfected Oat1 and Oat6-expressing cell lines, which could then be used to screen chemical libraries. SA3 (10-20% effort) involves initial ex vivo and in vivo "proof of concept" experiments. The proposed work will provide a paradigm for this type of screening of drugs and compounds that are transported not only across the olfactory epithelium but also the choroid plexus and brain capillary endothelium (including compounds that can be tagged for imaging of tumors and brain metabolism). We have begun preliminary experiments with NIH Roadmap funded facilities (Scripps) with the long-term goal of developing a synergistic high throughput virtual and wet lab screening strategy. All the expertise for completion of the aims exists within the PI's group. While the emphasis is on screening for CNS active drugs, it is to be emphasized that the proposal is also highly relevant to drug design for elimination via the kidney and liver. This proposal seeks to develop the basis for a virtual and wet lab high throughput screening system for drugs delivered to the central nervous system, including those useful for CNS tumors, CNS HIV infection and neuropsychiatric disorders. The goal is to identify highly specific substrates for the nasally expressed organic anion transporters, Oat1 and Oat6, thereby allowing for efficient intranasal delivery and transport of drugs to the CNS.

描述（由申请人提供）：该提案最终旨在为递送至中枢神经系统（CNS）的药物（包括对CNS肿瘤、CNS HIV感染和神经精神疾病有用的药物）的虚拟和湿实验室高通量筛选系统奠定基础。目的是鉴定我们发现的两种嗅觉表达的有机阴离子转运蛋白（Oats）Oat 6和Oat 1的高度特异性底物，从而允许药物经鼻递送至CNS。燕麦被认为是关键上皮组织-环境界面运动的限速步骤（例如，肾，肝）的许多常见药物（如，抗生素、抗病毒药、化疗药、NSAID、抗高血压药、利尿剂）。我们是第一个确定的原型，燕麦1，以及相关的转运蛋白，并在最近发表了燕麦1和燕麦3敲除，这有缺陷的有机阴离子转运在肾脏和脉络丛。由于Oat 6和Oat 1在所谓的鼻脑屏障和特定底物偏好处的表达，该途径对于用于鉴定CNS活性药物和探针的并行计算机模拟和湿实验室策略（和体内验证）是有吸引力的。假设Oat 6和Oat 1转运蛋白为药物经鼻给药至CNS提供了独特的转运机制，并且并行计算机模拟（SA 1）和湿实验室（SA 2）方法将证明具有协同作用，并更快地导致大量化合物的高通量虚拟和湿实验室筛选，以鉴定燕麦特异性集。然后，这些化合物可以作为潜在的线索，考虑可以经鼻递送的CNS作用药物，或者更可能的是，作为Oat 6和Oat 1介导的转运的一系列共识结构，可以考虑用于新药的设计或现有药物的衍生化与无效的CNS递送。SA 1（40-45%努力）的目标是使用受控分子动力学对Oat 6/Oat 1及其底物的相互作用进行计算机模拟，以便开始建立虚拟筛选的基础。SA 2（40- 45%努力）最初将继续湿实验室低中通量（非洲爪蟾卵母细胞运输）筛选潜在的Oat 6和Oat 1特异性底物，并开发一种采用稳定转染的Oat 1和Oat 6表达细胞系的高通量测定法，然后可用于筛选化学文库。SA 3（10-20%努力）涉及初始的离体和体内“概念验证”实验。拟议的工作将为这种类型的药物和化合物的筛选提供一个范例，这些药物和化合物不仅可以穿过嗅上皮，还可以穿过脉络丛和脑毛细血管内皮（包括可以标记用于肿瘤和脑代谢成像的化合物）。我们已经开始与NIH路线图资助的设施（Scripps）进行初步实验，长期目标是开发协同高通量虚拟和湿实验室筛选策略。完成目标的所有专业知识都存在于PI的团队中。虽然重点是筛选CNS活性药物，但需要强调的是，该提案也与通过肾脏和肝脏消除的药物设计高度相关。该提案旨在为递送至中枢神经系统的药物的虚拟和湿实验室高通量筛选系统开发基础，包括那些对CNS肿瘤、CNS HIV感染和神经精神疾病有用的药物。目的是确定鼻内表达的有机阴离子转运蛋白Oat 1和Oat 6的高度特异性底物，从而允许有效的鼻内递送和药物转运至CNS。