RESPIRATORY SYNCYTIAL VIRUS EFFICACY STUDY IN AFRICAN GREEN MONKEYS

非洲绿猴呼吸道合胞病毒功效研究

• 批准号: 8173023
• 负责人: VICKI L TRAINA-DORGE
• 金额: $ 6.18万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8173023
• 关键词: Animals; Anti-Inflammatory Agents; Anti-inflammatory; Antiviral Agents; Autopsy; Blood specimen; Body Temperature; Bronchoalveolar Lavage; Cercopithecus pygerythrus; Chemistry; Clinical; Collaborations; Computer Retrieval of Information on Scientific Projects Database; Continuous Intravenous Infusion; Control Groups; Detection; Dose; Euthanasia; Evaluation; Exposure to; Funding; Grant; Hour; Human Resources; IL8 gene; Infection; Inflammatory; Infusion procedures; Institution; Interleukin-6; Interleukins; Intravenous infusion procedures; Kinetics; Laboratories; Lung; Monitor; Nature; Nose; Pharmacologic Substance; Pharyngeal structure; Plasma; Primates; Property; Prophylactic treatment; RNA; Research; Research Personnel; Resources; Respiratory Syncytial Virus Infections; Respiratory syncytial virus; Sampling; Site; Source; Swab; Testing; Therapeutic; Trachea; Treatment Efficacy; United States National Institutes of Health; Viral Load result; Virus; Virus Replication; White Blood Cell Count procedure; aqueous; arm; drug distribution; food consumption; nonhuman primate; prevent; prophylactic; response; success; treatment effect; viral RNA

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. Two studies were conducted in African green monkeys (AGM) to test a proprietary compound (Cpd) formulated by Tibotec Pharmaceuticals against respiratory syncytial virus (RSV) in nonhuman primates. 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. The first study was conducted to test the prophylactic and therapeutic efficacy of this compound by continuous intravenous infusion in RSV infected primates at a plasma level of 50 ng/mL. Fifteen AGMs were divided into three groups: 1) the prophylactic (Px50) arm, received 0.033 mg Cpd- 4% aqueous Captisol¿ 24 hours before infection for 10 days, 2) the therapeutic (Tx50) arm, received the same dose at 24 hours post infection for eight days, and 3) the control arm, received vehicle - 4% aqueous Captisol¿ 24 hours after infection for eight days. All fifteen AGMs were challenged with 104 plaque forming units (pfu) per mL RSV A2 intranasally and intratracheally. The AGMs were necropsied 14 days after challenge. Samples of the site of infusion, trachea, and lung were taken for histopathological evaluation. Viral load was determined by means of PCR and culture of nasal and throat swabs and bronchoalveolar lavages (BAL) taken before and during the infection. Blood samples were taken to confirm target exposure and to evaluate the effects of treatment on hematologic, immunological, and plasma chemistry parameters. Furthermore, the animals were observed daily, and their food consumption and body temperatures were monitored. The primary endpoint, RSV RNA viral loads, demonstrated that the prophylactic treatment with compound not only reduced the peak viral load by 1.5 log10 RNA copies per mL but also delayed the peak for two days. The therapeutic treatment only reduced the peak viral load by 1.5 log10 RNA copies per mL with no delay in the peak. From the secondary endpoints, the only difference observed was in the IL-6 levels, which were lower in the prophylactic and therapeutic groups than in the control groups. In conclusion, both prophylactic and therapeutic treatment at a continuous plasma level of 50 ng/mL for 10 and 8 days following infection, respectively, were effective in reducing the peak viral load, but not in preventing RSV infection. A third and final study was conducted to test the prophylactic efficacy of this same compound in RSV infected primates at plasma levels of 5 and 500 ng/mL. Twelve AGMs were divided into three groups: 1) Prophylactic 5 (Px5), received an intravenous infusion of 0.0033 mg / mL Cpd - 4% aqueous Captisol¿ 72 hours before infection for 16 days, 2) Prophylactic 500 (Px500), received an intravenous infusion of 0.33 mg/ml Cpd - 4% aqueous Captisol¿ 72 hours before infection for 16 days, 3) vehicle control group, received an intravenous infusion of 4% Captisol¿ 72 hours before infection for 16 days. All twelve AGMs were challenged with 104 plaque forming units (pfu) per mL RSV A2 intranasally and intratracheally on day 0. The AGMs were necropsied 13 days after challenge. Clinical monitoring was as stated above for the earlier study. Viral loads as determined by PCR showed a similar kinetics in BAL and throat swabs for each of the groups. By culture and PCR, all Px500 animals had undetectable RNA viral loads in BAL and throat swabs rinses except AGM Y459, which showed positive PCR responses in the throat swab rinses and BAL shortly before euthanasia, which was probably due to insufficient distribution of the drug. The Px5 animals did not show a decrease but instead displayed a delayed viral load peak in the BAL and throat swab rinses. Interleukin- (IL-) 6 and IL-8 were suppressed only in the Px500 group, and slightly decreased in the Px5 group compared to the control group. White blood cell (WBC) counts in BAL appeared reduced in the Px500 group compared to the Px5 and vehicle control groups. Histological examination showed very pronounced changes predominantly of an inflammatory nature in the vehicle control group and Px5 group and much less pronounced changes in the Px500 group. In summary, prophylactic treatment of AGMs with a target plasma level of 500 ng/mL for 16 days resulted in the suppression of RSV replication to a level below the lower limit of detection, a clear treatment success. Prophylactic treatment of AGMs with a target plasma level of 5 ng/mL in the same manner did not show an antiviral effect, but demonstrated certain anti-inflammatory properties.

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)中进行了两项研究，以测试Tibotec制药公司配制的一种专利化合物(CPD)在非人类灵长类动物中对抗呼吸道合胞病毒(RSV)的作用。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，而且对从动物身上提取的样本进行所有病毒学测试。第一项研究是通过对RSV感染灵长类动物持续静脉滴注50 ng/m L的血浆水平来检测该化合物的预防和治疗效果。15只巨噬细胞分为3组：预防性组(Px50)，感染前24小时给予0.033 mg CPD-4%水卡替索尔，连续10天；2)治疗组(TX50)，感染后24小时给予相同剂量，连续8天；所有15个AGM在鼻腔和气管内以每毫升RSV A2 104个空斑形成单位(PFU)激发。攻击后14天剖检AGM。取输液部位、气管和肺的标本进行组织病理学评价。感染前和感染期间分别取鼻咽拭子和支气管肺泡灌洗液(BAL)，用聚合酶链式反应和培养法检测病毒载量。采集血样以确认目标暴露，并评估治疗对血液学、免疫学和血浆化学参数的影响。此外，每天对这些动物进行观察，并监测它们的食物摄入量和体温。主要终点是RSV RNA病毒载量，结果表明，复方预防处理不仅使病毒载量峰值降低了1.5log10RNA拷贝/毫升，而且还将高峰推迟了两天。治疗仅使病毒载量峰值降低1.5log10RNA拷贝数/毫升，且峰值没有延迟。从次要终点来看，唯一的差异是IL-6水平，预防和治疗组低于对照组。总之，预防治疗和治疗治疗分别在感染后10天和8天持续维持50 ng/mL的血浆水平，可以有效地降低病毒载量峰值，但对预防RSV感染无效。 第三项也是最后一项研究是为了测试该化合物在血浆水平为5和500 ng/mL时对感染RSV的灵长类动物的预防效果。12只小鼠分为3组：预防5组(PX5)，感染前72小时静脉滴注0.0033 mg/mLCPD-4%卡替索尔，连续16天；2)预防500(PX500)，感染前72小时静脉滴注4%卡替索尔0.33 mg/mL16天；所有12个AGM在第0天经鼻腔和气管内以每毫升RSV A2 104个空斑形成单位(PFU)进行攻击。挑战后13天，解剖动物模型。临床监测如上所述，用于早期研究。通过聚合酶链式反应测定的病毒载量显示，每一组的BAL和咽拭子中的病毒载量的动力学相似。通过培养和PCR，除AGM Y459在安乐死前不久在咽拭子和BAL中呈阳性反应外，所有Px500动物的BAL和咽拭子冲洗液中均未检测到RNA病毒载量，这可能是由于药物分布不充分所致。PX5动物没有表现出下降，而是在BAL和咽拭子冲洗中显示出延迟的病毒载量峰值。IL-6和IL-8仅在Px500组受到抑制，而PX5组与对照组相比略有下降。与PX5和赋形剂对照组相比，Px500组BAL中的白细胞(WBC)计数似乎有所下降。组织学检查显示，赋形剂对照组和PX5组有非常显著的变化，主要是炎性变化，而PX500组的变化要小得多。总而言之，对目标血浆水平为500 ng/mL的急性呼吸道合胞病毒感染患者预防性治疗16天后，RSV复制被抑制到低于检测下限的水平，这是一个明显的治疗成功。以同样的方式预防性处理目标血药浓度为5 ng/m L的AGM并未显示出抗病毒作用，但表现出一定的抗炎特性。