FY20. RECEPTOR PROFILES - TASK ORDER NO. 75N95020F00159 POP: SEPTEMBER 30, 2020 - SEPTEMBER 29, 2021. N01DA-18-8937.

2020 财年。

• 批准号: 10285161
• 负责人: JACOB BODE
• 金额: $ 35.91万
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-30 至 2021-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10285161
• 关键词: Agonist; Binding; Biological; Biological Assay; Cells; Contractor; Contracts; Cytochrome P450; Data; Disease; Drug Addiction; Enzymes; Evaluation; Exposure to; In Vitro; Ligands; Measures; Medical; Micronucleus Tests; National Institute of Drug Abuse; Pharmaceutical Preparations; Ships; Site; Specific qualifier value; Structure; Testing; Therapeutic; Time; Toxic effect; Toxicology; United States; Work; addiction; biological systems; follow-up; novel; pre-clinical; programs; receptor; receptor binding; screening

Under Task 1, the Contractor shall receive up to twenty-nine (29) test compounds to be evaluated for receptor profile screening. The Contractor shall screen these compounds at two concentrations (100 nM and 10,000 nM) in duplicate (four (4) wells per target). The Contractor shall screen for binding at the biological targets listed in the profile unless otherwise requested by the Project Officer. Under Task 2, the Contractor shall perform up to eighty (80) evaluations of receptor binding, functional activity, or other assays. The exact number and type of assays to be performed usually cannot be specified until after Task 1 work is completed but may include both binding and functional activity measures at receptor, transporter, and enzyme sites. Each evaluation shall consist of two replications of eight (8) concentrations using duplicate wells in each replication (total of 32 wells per assay) and will determine the IC50/Ki or functional activity at the target receptor. A screening assay at 100 and 10,000 nM in duplicate (4 wells) or other concentrations as specified by the NIDA COR may be performed prior to study to help determine the best range of concentrations to be tested. Functional assays will include both agonist and antagonist function unless otherwise specified by the NIDA COR. Under Task 3, the Contractor shall perform up to sixty (60) toxicity assays including Cytochrome P450 profiles, hERG channel assays, Ames mutagenicity assays and P-gp substrate assays. Each Cyp450 profile shall consist of testing duplicate wells at 100 and 10,000 nM for six (6) Cyp450 enzymes specified in the Statement of Work (total of twenty-four (24) wells). A Ki/IC50 evaluation will be performed at each site where the profile shows at least twenty-five percent (25%) binding inhibition at either concentration (a “hit”). Each follow-up evaluation shall consist of two replications of eight (8) concentrations using duplicate wells in each replication (total of 32 wells per assay). The hERG channel assay will consist of five (5) concentrations and triplicate cells (total of 15 data points). The micronucleus assay will consist of 8 concentrations tested in triplicate wells with and without exposure to S9 (48 wells total) unless otherwise directed by the NIDA COR. Under Task 6, the Contractor shall purchase up to five (5) compounds for testing at the direction of the NIDA COR. The NIDA COR will have determined previously that the compounds could not be purchased in the United States and shipped to the Contractor in a reasonable time frame. Money to be used for this Task will be subtracted from the amount of money allocated to other Tasks and the number of assays performed under such Tasks reduced.

在任务1项下，承包商应接收多达二十九（29）种测试化合物进行受体谱筛选评估。承包商应以两种浓度（100 nM和10,000 nM）筛选这些化合物，一式两份（每个目标四(4)孔）。除非项目官员另有要求，承包商应在简介中列出的生物目标处进行筛选，以确定是否存在结合。在任务2项下，承包商应执行多达80项受体结合、功能活性或其他分析评估。要进行的测定的确切数量和类型通常要在任务1工作完成后才能指定，但可能包括受体、转运体和酶位点的结合和功能活性测量。每次评估应包括8(8)浓度的两次重复，每次重复使用重复孔（每次分析共32孔），并将确定IC50/Ki或目标受体的功能活性。在研究之前，可以在100和10,000 nM（4孔）或NIDA COR指定的其他浓度下进行筛选试验，以帮助确定要测试的最佳浓度范围。功能测试将包括激动剂和拮抗剂功能，除非NIDA COR另有规定。根据任务3，承包商应执行多达60项毒性测试，包括细胞色素P450谱、hERG通道测试、Ames诱变性测试和P-gp底物测试。每个Cyp450剖面应包括工作说明书中指定的六(6)种Cyp450酶在100和10,000 nM下的测试重复孔（总共24（24）个孔）。Ki/IC50评估将在每个位点进行，其中在任何浓度下，谱显示至少25%（25%）的结合抑制（“命中”）。每次后续评估应包括8个浓度的两次重复，每次重复使用重复孔（每次分析共32个孔）。hERG通道试验将包括5个浓度和3个复制细胞（总共15个数据点）。除非NIDA COR另有指示，否则微核分析将包括在三口井中进行8种浓度的测试，有或没有暴露于S9（总共48口井）。在任务6项下，承包商应按照NIDA COR的指示购买最多五(5)种化合物进行测试。NIDA COR将事先确定这些化合物无法在美国购买并在合理的时间范围内运送给承包商。用于此任务的资金将从分配给其他任务的资金中扣除，并减少在这些任务下执行的分析次数。