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.

该副本是使用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这是调查员的机构。 在非洲绿色猴子（AGM）中进行了两项研究，以测试由Tibotec Pharmaceuticals针对呼吸综合病毒（RSV）在非人类隐私中提出的专有化合物（CPD）。这些研究是与动物和兽医在其设施中与生物试验公司合作进行的。我的实验室不仅要提供病毒挑战库存，而且还对动物采集的样品进行了所有病毒学测试。进行了第一项研究，以测试该化合物的预防和治疗效率，通过在50 ng/mL的血浆水平下连续静脉输注感染的PRIME中连续静脉输注。 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 therapy (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.所有15个AGM均受到每ML RSV A2的104个斑块形成单元（PFU）的挑战，并进行了气管内。挑战后14天后，AGM被尸检。取输注，气管和肺部的样本进行组织病理学评估。病毒载量是通过PCR和鼻拭子的培养以及在感染前后进行的支气管肺泡灌洗（BAL）确定的。采集血液样本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，每天都观察到动物，并监测其食物消耗和体温。主要终点RSV RNA病毒载荷表明，使用化合物的预防性处理不仅使峰值病毒载荷降低了1.5 log10 RNA拷贝，而且还将峰延迟了两天。治疗仅将峰值病毒负荷降低1.5 log10 RNA拷贝，而峰值无延迟。从次要终点来看，观察到的唯一差异是在IL-6水平中，在预防和治疗组中比对照组低。总之，在感染后的50 ng/mL的连续血浆水平为50 ng/mL的连续血浆水平上，感染后10和8天有效地减少了峰值病毒载量，但不能防止RSV感染。 进行了第三项也是最后一项研究，以测试RSV感染的Prime中同一化合物的预防效率，在500 ng/ml的血浆水平下。将十二个AGM分为三组：1）预防5（PX5），静脉输注为0.0033 mg/ml CPD -4％感染16天前72小时，持续时间为16天，2）预防500（PX500）。感染16天，3）媒介物对照组，在感染前72小时接受静脉输注4％CAPTERISOL。16天。所有十二个AGM均在第0天，每毫升RSV A2鼻内和弹药内挑战104个斑块形成单位（PFU）。挑战后13天，AGM被杀死。临床监测如上所述。由PCR确定的病毒载荷显示了每个组的BAL和喉咙拭子的动力学。通过培养和PCR，除AGM Y459以外，所有PX500动物在BAL和喉咙拭子路线中都有无法检测到的RNA病毒载荷，该路线在喉咙拭子路线中显示出阳性的PCR反应，而在安乐死前不久，这可能是由于药物分布不足所致。 PX5动物没有显示下降，而是在BAL和喉咙拭子中显示出延迟的病毒载荷峰。白介素 - （IL-）6和IL-8仅在PX500组中被抑制，与对照组相比，PX5组中的PX5组略有改进。与PX5和媒介物对照组相比，PX500组的BAL中白细胞（WBC）计数降低。组织学检查显示，媒介物对照组和PX5组的炎症性质的变化非常明显，而PX500组的变化却不那么明显。总之，对目标血浆水平为500 ng/ml的AGM 16天的预防治疗导致RSV复制抑制至低于检测下限的水平，这是明显的治疗成功。以相同方式对目标血浆水平为5 ng/mL的AGM的预防性治疗没有显示抗病毒作用，而是表现出某些抗炎特性。