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的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 在非洲绿色猴（AGM）中进行了两项研究，以检测Tibotec Pharmaceuticals配制的专利化合物（Cpd）在非人灵长类动物中抗呼吸道合胞病毒（RSV）的作用。这些研究与Bioqual，Inc.合作进行。与动物和兽医人员在他们的设施。我的实验室不仅要提供病毒攻毒储备液，还要对从动物身上采集的样本进行所有病毒学检测。 进行第一项研究以测试该化合物通过在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的预防和治疗功效。将15个AGM分为三组：1）预防性（Px 50）臂，在感染前24小时接受0.033 mg Cpd- 4% Captisol水溶液，持续10天，2）治疗性（Tx 50）臂，在感染后24小时接受相同剂量，持续8天，和3）对照臂，接受载体-4% Captisol水溶液，感染后24小时，持续8天。 所有15个AGM均用104空斑形成单位（pfu）/mL RSV A2鼻内和鼻内攻毒。 攻毒后14天对AGM进行尸检。 采集输注部位、气管和肺样本进行组织病理学评价。 病毒载量通过PCR和培养的鼻和咽拭子和支气管肺泡灌洗液（BAL）采取的感染前和期间。 采集血样以确认目标暴露量，并评价给药对血液学、免疫学和血浆化学参数的影响。 此外，每天观察动物，并监测其摄食量和体温。 主要终点RSV RNA病毒载量证明，化合物预防性治疗不仅使病毒载量峰值降低了1.5 log 10 RNA拷贝/mL，而且使峰值延迟了2天。 治疗性处理仅使峰值病毒载量降低1.5 log 10 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10天和8天分别以50 ng/mL的连续血浆水平进行预防性和治疗性治疗，可有效降低病毒载量峰值，但不能预防RSV感染。 进行第三和最后一项研究以测试该相同化合物在5和500 ng/mL血浆水平下在RSV感染的灵长类动物中的预防功效。 12例AGM分为3组：1）预防组5（Px 5），感染前72 h静脉滴注0.0033 mg / mL Cpd - 4% Captisol水溶液，共16 d; 2）预防组500（Px 500），感染前72 h静脉滴注0.33 mg/ml Cpd - 4% Captisol水溶液，共16 d对照组于感染前72 h静脉滴注4% Captisol <$16 d。 在第0天，所有12个AGM均用104空斑形成单位（pfu）/mL RSV A2鼻内和鼻内攻毒。 攻毒后13天对AGM进行尸检。 临床监测与上述早期研究相同。 通过PCR测定的病毒载量在各组的BAL和咽拭子中显示出相似的动力学。 通过培养和PCR，除AGM Y 459外，所有Px 500动物的BAL和咽拭子冲洗液中均未检出RNA病毒载量，AGM Y 459在人道处死前不久的咽拭子冲洗液和BAL中显示阳性PCR反应，这可能是由于药物分布不足所致。 Px 5动物未显示降低，但在BAL和咽拭子冲洗液中显示延迟的病毒载量峰值。 与对照组相比，仅在Px 500组中抑制了白细胞介素-（IL-）6和IL-8，并且在Px 5组中略微降低。与Px 5和溶剂对照组相比，Px 500组BAL中的白色血细胞（WBC）计数似乎降低。组织学检查显示，在溶剂对照组和Px 5组中主要是炎症性质的非常明显的变化，而在Px 500组中变化不太明显。 总之，以500 ng/mL的目标血浆水平预防性治疗AGM 16天，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。 以相同的方式对目标血浆水平为5 ng/mL的AGM进行预防性治疗未显示出抗病毒作用，但显示出一定的抗炎特性。