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.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 在非洲绿猴 (AGM) 中进行了两项研究，以测试 Tibotec Pharmaceuticals 配制的专利化合物 (Cpd) 对非人类灵长类动物呼吸道合胞病毒 (RSV) 的抵抗力。这些研究是与 Bioqual, Inc. 合作进行的，动物和兽医人员在其工厂进行。我的实验室不仅要提供病毒挑战库存，还要对从动物身上采集的样本进行所有病毒学测试。 第一项研究是通过在 RSV 感染的灵长类动物中以 50 ng/mL 的血浆水平连续静脉输注来测试该化合物的预防和治疗功效。 15 个 AGM 分为三组：1) 预防 (Px50) 组，在感染前 24 小时接受 0.033 mg Cpd- 4% Captisol 水溶液，持续 10 天；2) 治疗 (Tx50) 组，在感染后 24 小时接受相同剂量，持续 8 天；3) 对照组，在感染后 24 小时接受媒介物 - 4% Captisol 水溶液，持续 8 天。 所有 15 个 AGM 均接受鼻内和气管内每 mL RSV A2 104 个噬菌斑形成单位 (pfu) 的挑战。 攻击后14天对AGM进行尸检。 采集输注部位、气管和肺部样本用于组织病理学评估。 通过 PCR 以及感染前和感染期间采集的鼻咽拭子和支气管肺泡灌洗液 (BAL) 培养来确定病毒载量。 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，每天观察动物，并监测它们的食物消耗和体温。 主要终点 RSV RNA 病毒载量表明，使用化合物进行预防性治疗不仅将峰值病毒载量降低了 1.5 log10 RNA 拷贝/mL，而且还将峰值延迟了两天。 治疗仅将峰值病毒载量降低了 1.5 log10 RNA 拷贝/mL，峰值没有延迟。 从次要终点来看，观察到的唯一差异是 IL-6 水平，预防组和治疗组的 IL-6 水平低于对照组。 总之，感染后连续 10 天和 8 天持续血浆浓度为 50 ng/mL 的预防性治疗和治疗性治疗可有效降低病毒载量峰值，但不能预防 RSV 感染。 第三项也是最后一项研究是为了测试该相同化合物在血浆浓度为 5 ng/mL 和 500 ng/mL 时对 RSV 感染的灵长类动物的预防功效。 12 名 AGM 分为三组：1) 预防性 5 (Px5)，感染前 72 小时接受静脉输注 0.0033 mg/mL Cpd - 4% Captisol 水溶液，持续 16 天；2) 预防性 500 (Px500)，感染前 72 小时接受静脉输注 0.33 mg/ml Cpd - 4% Captisol 水溶液。 感染16天，3)载体对照组，在感染前72小时接受静脉输注4% Captisol¿，持续16天。 在第 0 天，所有 12 个 AGM 均在鼻内和气管内用每 mL RSV A2 104 个噬菌斑形成单位 (pfu) 进行攻击。攻击后 13 天对 AGM 进行尸检。 临床监测如上文早期研究所述。 PCR 测定的病毒载量显示，各组的 BAL 和咽拭子中的病毒载量具有相似的动力学。 通过培养和 PCR，所有 Px500 动物在 BAL 和咽喉拭子冲洗液中均检测不到 RNA 病毒载量，但 AGM Y459 除外，AGM Y459 在安乐死前不久在咽喉拭子冲洗液和 BAL 中显示出阳性 PCR 反应，这可能是由于药物分布不足所致。 Px5 动物没有表现出下降，而是在 BAL 和咽喉拭子冲洗液中表现出延迟的病毒载量峰值。 白细胞介素-(IL-)6和IL-8仅在Px500组中受到抑制，并且与对照组相比，Px5组中略有下降。与 Px5 和媒介物对照组相比，Px500 组 BAL 中的白细胞 (WBC) 计数似乎有所减少。组织学检查显示在媒介物对照组和Px5组中主要是炎症性质的非常明显的变化，而在Px500组中则显示不太明显的变化。 总之，以 500 ng/mL 的目标血浆水平进行 AGM 预防性治疗 16 天，可将 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浓度为 5 ng/mL 的 AGM 进行预防性治疗，并未显示出抗病毒作用，但显示出一定的抗炎特性。