Core A

核心B的总体目标是为IPCP提供药物相互作用的药敏试验和评价 程序. BBI Biotech对HIV-1进行了许多药物敏感性试验，这些试验有助于 一种新的抗逆转录病毒药物PA 457的开发，刚刚完成了一个成功的I期临床试验。最初 Core将筛选5种NK-1 R拮抗剂（阿瑞匹坦、CJ-12255、CJ-96345、RP-67,580和L733060），用于抗逆转录病毒治疗。 活性和细胞毒性。有效化合物，即，抗逆转录病毒药物比例可接受者 将在项目1和2以及核心B中进一步表征。表征 抗病毒反应的广度，除了阿瑞匹坦（计划用于初始动物研究， 临床试验），将针对由M型、O型、HIV-2原代分离株组成的一组25个HIV分离株进行检测 和耐药菌株。将确定每种分离株的50%有效剂量（EC 50）。因为大多数抗逆转录病毒药物 治疗包含多种药物，阿瑞匹坦和其他先导化合物将分别进行药物 与不同种类的抗逆转录病毒药物的相互作用。用于相互作用测试的药物将与HIV-1一起培养 (R5和X4病毒）。HIV p24产生的减少将用于中位效应 计算EC 50的方程和指示协同作用或拮抗作用的组合指数（CI）。为了评价 体外抗性诱导，THP-1细胞将与HIV-1 Ba-L和低剂量阿瑞匹坦一起培养。药物 每月进行一次敏感性试验，以寻找正在产生的耐药性。如果EC 50增加，病毒 信封将被测序，以确定共受体结合区和病毒的变化，并将细胞发送到项目1和2， 分别为进一步研究。为了确定阿瑞匹坦耐药性是否可以在体内发展，分离自 将每隔一个月对项目3中用阿瑞匹坦处理的猴进行测定，并测定EC 50。关联 治疗效果，患者病毒群、入组前分离株、治疗结束和一个 将测定项目4中阿瑞匹坦治疗后1个月的受试者对阿瑞匹坦和共受体使用的敏感性。如果 如果共受体使用或药物敏感性发生变化，则将在治疗前和治疗后对信封进行测序 处理分离物以鉴定抗性标记物。核心B提供的数据将决定宽度和稳定性 阿瑞匹坦在体内和体外的抗病毒作用，并表征其他NK-1 R拮抗剂与抗逆转录病毒 未来的研究活动。

The overall goal of Core B is to provide drug susceptibility testing and evaluation for drug interactions for the IPCP program. BBI Biotech has performed numerous drug susceptibility assays for HIV-1, which have contributed to the development of a novel anti-retroviral drug, PA457, that just completed a successful Phase I clinical trial. Initially, the Core will screen five NK-1R antagonists (aprepitant, CJ-12255, CJ-96345, RP-67,580, and L733060), for anti-retroviral activity and cytotoxicity in PBMC using HIV-1 Ba-L. Effective compounds, i.e., those with acceptable ratio of antiretroviral activity to cytotoxicity, will be further characterized in Projects 1 and 2, and Core B. To characterize the breadth of the antiviral response, the lead compounds, in addition to aprepitant (planned for initial animal study and clinical trial), will be tested against a panel of 25 HTV isolates consisting of type M, type O, HIV-2 primary isolates and drug resistant isolates. The 50% Effective Dose (EC50) will be determined against each isolate. Because most antiretroviral therapies contain multiple drugs, aprepitant and the other lead compounds will each be tested for drug interactions against different classes of anti-retrovirals. Drugs to be tested for interactions will be cultured with HIV-1 (R5 and X4 viruses) in PBMC together or individually. Reduction in HIV p24 production will be used in the medianeffect equation to calculate the EC50s and combination indices (CI) to indicate synergism or antagonism. To evaluate in vitro resistance induction, THP-1 cells will be cultured with HIV-1 Ba-L and low doses of aprepitant. Drug susceptibility assays will be performed once a month to look for developing resistance. If the EC50 increases, the viral envelope will be sequenced to identify changes in co-receptor binding area and virus and cells sent to Projects 1 and 2, respectively, for further study. To determine whether aprepitant resistance can develop in vivo, SFV isolated from monkeys treated with aprepitant in Project 3 will be assayed every other month and EC50 determined. To associate treatment effects with possible changes in patient viral population, isolates from pre-entry, end of treatment, and one month after aprepitant treatment from Project 4 will be assayed for susceptibility to aprepitant and co-receptor usage. If there are changes in co-receptor usage or drug susceptibility the envelope will be sequenced in pretreatment and post treatment isolates to identify resistance markers. The data provided by Core B will determine the breadth and stability of the anti-viral effects of aprepitant both in vivo and in vitro and characterize additional NK-1R antagonists with antiretroviral activities for future studies.