RESPIRATORY SYNCYTIAL VIRUS EFFICACY STUDY IN AFRICAN GREEN MONKEYS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 在非洲绿猴(AGM)中进行了一系列三项研究，以测试由Tibotec制药公司配制的专利化合物TMC 353121在非人类灵长类动物中对抗呼吸道合胞病毒。在之前的研究中，这种化合物，一种RSV融合抑制剂，在较小的动物身上被证明对RSV有疗效。这些研究是与Bioqual公司合作进行的，动物和兽医人员在他们的设施中进行。我的实验室不仅提供病毒挑战库存，而且在三项研究的每一项期间对动物样本进行所有病毒学测试。我们对48个AGM进行了血清学测试，以预先筛选和识别36只RSV血清阴性动物用于研究。第一项研究，NC282，是一项感染研究，使用了6个AGM。我们分别以1x10e3pfu/ml和1x10e4pfu/ml两种浓度制备了37、1ml的RSV毒株，并将它们冷冻保存用于整个研究。动物分为两组，每组分别经鼻腔和气管内注射10e3pfu/ml(Gp.1)，另一组给予10e4pfu/ml(Gp.2)。每天采集鼻和喉部样本，每两天采集一次支气管肺泡灌洗液样本，并通过空斑试验检测病毒载量，以监测动物是否有呼吸道合胞病毒感染的临床症状。两组病毒载量均在第6天达到高峰，咽喉分别为3.7e2pfu/ml和1.4e3pfu/ml，BAL分别为2e2pfu/ml和3.7e2pfu/ml，Gps1和2分别达到峰值。这些结果证实了该模型，并决定将10e4pfu/ml的接种剂量用于随后的两项疗效研究。进行了两次持续静脉滴注该化合物的疗效研究。研究2有15只动物被分成三组，使用GPS。实验3组12只动物随机分为3组，每组12只，分别以5 ng/ml或500 ng/ml的血浆浓度注射TMC-353121，GP-3为对照组。动物被预先准备好穿上夹克和系绳系统，接受了植入导管的手术，然后开始输液。动物研究最近已经完成，但我们仍在进行斑块和抗体分析，并收集数据。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. A series of three studies were conducted in African green monkeys (AGM) to test a proprietary compound, TMC 353121, formulated by Tibotec Pharmaceuticals against RSV in nonhuman primates. In previous studies, the compound, an RSV fusion inhibitor was shown to have efficacy against RSV in vivo in smaller animals. These studies were performed in collaboration with Bioqual, Inc. with animals and veterinary personnel at their facility. My laboratory was to provide not only virus challenge stock but performed all virological testing of samples taken from the animals during each of the three studies. We performed serological testing of 48 AGMs to prescreen and identify 36 RSV seronegative animals to be used for the studies. The first study, NC282, was an infection study and utilized 6 AGM. We prepared 37, 1ml vials of RSV strain at each of two concentrations of 1x10e3pfu/ml and 1x10e4pfu/ml and cryopreserved them for the entire study. The animals were divided into two groups with each group receiving either 10e3pfu/ml (Gp.1) by intranasal and intratrachael administration and the other receiving 10e4pfu/ml (Gp.2) by similar administration. Animals were monitored for clinical signs of RSV infection with nose and throat samples taken daily and bronchoalveolar lavage samples taken every two days and tested for virus loads by plaque assay. Peak viral loads were reached in both groups by day 6 in the throat with 3.7e2 pfu/ml and 1.4 e3pfu/ml and in the BAL with 2e2 pfu/ml and 3.7e2 pfu/ml and for Gps1 and 2 respectively. These results confirmed the model and the decision to utilize the 10e4pfu/ml inoculation dose for the two subsequent efficacy studies was made. Two efficacy studies with the continuous intravenous infusion of the compound were performed. Study 2 had 15 animals divided into three groups with Gps. 1 and 2, therapeutic and prophylactic arms with TMC 353121 was administered at a plasma level of 50ng/ml and Gp 3 the vehicle control arms; and Study 3 had 12 animals divided into three groups with Gps 1 and 2 therapeutic arms only with TMC 353121 administered at plasma levels of either 5ng/ml or 500ng/ml and Gp 3 a vehical control group. Animals were preconditioned to wear the jacket and tethering system, underwent surgery for implanting the catheter, and infusions begun. The animal studies have recently been completed, however, we are still conducting plaque and antibody assays and compiling data.