Project 2: PREVENT Preclinical safety and efficacy studies

项目2：预防临床前安全性和有效性研究

• 批准号: 9276592
• 负责人: KENNETH E PALMER
• 金额: $ 12.8万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9276592
• 关键词: Address; Adult; Affect; Affinity; Anal Sex; Animals; Anti-Retroviral Agents; Antiviral Agents; Anus; Attention; Awareness; Binding; Biological Assay; Biopsy; CCR5 gene; CD209 gene; Carbopol; Cell surface; Cells; Clinical; Clinical Research; Clinical Trials; Colorectal; Communities; Community Health; Control Animal; Data; Data Set; Detection; Development; Dose; Drug Kinetics; Effectiveness; Environment; Enzyme-Linked Immunosorbent Assay; Epithelial; Epithelial Cells; Exposure to; Female; Flow Cytometry; Formulation; Gel; Gene Chips; Gene Expression; Goals; HIV; HIV-1; Health; Hepatitis C virus; Heterosexuals; Human; Human Herpesvirus 2; Human Volunteers; Immune; Immune response; Immunity; Immunoassay; Immunohistochemistry; Incidence; Infection; Laboratories; Lectin; Link; Liquid substance; Local Microbicides; Lymphocyte; Macaca; Macaca mulatta; Mass Spectrum Analysis; Measures; Metagenomics; Methods; Modeling; Mucous Membrane; Mus; Outcome; Pharmaceutical Preparations; Pharmacodynamics; Phenotype; Placebo Control; Placebos; Polysaccharides; Population; Prevention; Proteins; Proteomics; Rectum; Reporting; Research Design; Safety; Sexual Transmission; Staging; Structure; Surface; Systems Biology; Techniques; Testing; Tissues; Trauma; Vagina; Viral; Virus Diseases; Virus Receptors; Virus Shedding; Woman; base; cohort; crosslink; design; experience; immunotoxicity; in vivo; inflammatory marker; langerin; male; men who have sex with men; microbial community; microbicide; microbiome; microbiota; novel; placebo controlled study; preclinical efficacy; preclinical safety; prevent; product development; protective efficacy; rRNA Genes; rectal; rectal microbicide; research clinical testing; response; safety testing; simian human immunodeficiency virus; transcriptomics; transmission process; trial design; viral transmission

The antiviral lectin Griffithsin (GRFT) has broad spectrum activity against human immunodeficiency viruses (HIV) types 1 and 2, herpes simplex virus type 2 (HSV-2) and hepatitis C virus (HCV) (1-4). Griffithsin-based topical microbicide formulations offer the potential for use as a multipurpose, non- antiretroviral based method to prevent HIV-1 and epidemiologically linked infections. The Project 2 objective is to show that GRFT containing rectal microbicide products are safe and effective in the Rhesus macaque model of rectal HIV transmission. These studies are designed to be complementary to the human clinical studies proposed in Project 3. We have three specific aims: Aim 1 is to evaluate the safety, pharmacokinetics and pharmacodynamics (PK/PD) of GRFT carbopol gel. We will conduct a three stage placebo controlled PK/PD study in adult male Rhesus macaques with the GRFT rectal gel formulation and matched placebo gel supplied by Core B. The proposed macaque study is designed to address specific translational questions concerning immunotoxicity and health of the mucosal environment after GRFT exposure. Favorable stage 1 outcomes include detection of active HIV-1 inhibitory concentrations of GRFT in rectal fluids and acceptable clinical safety metrics, which allows progression to the efficacy Stage 2. Using a cohort of GRFT treatment experienced animals will allow us to determine whether pre-existing immunity to GRFT affects product efficacy. Stage 3 will test the GRFT and placebo products in SHIV infected animals, where we will compare amounts of virus shed in experimental versus control animals, providing an additional safety metric. Aim 2 is to assess the impact of GRFT treatment on the mucosal immune environment. We will use multiplexed immunoassays and gene expression arrays to measure markers of inflammatory immune responses. The highly sensitive immunohistochemistry (IHC) techniques refined in Prof. Kristina Broliden's laboratory will be applied to compare the number, distribution and activation state of HIV target cells and expression of HIV receptor targets CD4, CCR5, DC-SIGN and Langerin plus markers of epithelial integrity in rectal biopsies from animals treated with GRFT active drug or matched placebo. Flow cytometry analyses will also enumerate and phenotype lymphocytes in biopsy tissues. Aim 3 is to assess the impact of GRFT treatment on the mucosal environment in the rectum using a systems biology approach that utilizes modern mass spectrometry proteomics assays to compare tissues and rectal secretions from animals that receive active drug versus placebo, and to evaluate the effect of GRFT treatment on the rectal microbiota of treated animals.

抗病毒凝集素Griffithsin（GRFT）具有广谱抗人类免疫缺陷的活性 病毒（HIV）1型和2型、单纯疱疹病毒2型（HSV-2）和丙型肝炎病毒（HCV）（1-4）。 基于Griffithsin的局部杀微生物剂制剂提供了用作多用途、非生物相容性杀微生物剂的潜力。 以抗逆转录病毒为基础的方法来预防HIV-1和流行病学相关感染。项目2 目的是证明含有GRFT的直肠杀微生物剂产品在治疗直肠癌中是安全有效的。 恒河猴直肠HIV传播模型。这些研究旨在 补充项目3中提出的人体临床研究。我们有三个具体目标： 目的1：评价GRFT的安全性、药代动力学和药效学（PK/PD） 卡波姆凝胶我们将在成年雄性恒河猴中进行一项三阶段安慰剂对照PK/PD研究 猕猴用核心B提供的GRFT直肠凝胶制剂和匹配的安慰剂凝胶。的 拟议的猕猴研究旨在解决具体的翻译问题， GRFT暴露后粘膜环境的免疫毒性和健康。有利阶段1 结果包括检测直肠液中GRFT的活性HIV-1抑制浓度， 可接受的临床安全性指标，允许进展至疗效阶段2。使用队列 GRFT治疗经验丰富的动物将使我们能够确定是否预先存在的免疫力， GRFT影响产品功效。第3阶段将在SHIV感染者中测试GRFT和安慰剂产品 动物，我们将比较实验动物与对照动物中的病毒脱落量， 提供了额外的安全度量。目的2是评估GRFT治疗对 粘膜免疫环境我们将使用多重免疫分析和基因表达阵列 来测量炎症免疫反应的标志物。高度敏感的 将应用Kristina Broliden教授实验室改进的免疫组织化学（IHC）技术 比较HIV靶细胞的数量、分布、活化状态和HIV的表达 受体靶向CD 4、CCR 5、DC-SIGN和Langerin加上直肠上皮完整性标志物 来自用GRFT活性药物或匹配的安慰剂治疗的动物的活组织检查。流式细胞术分析 还将对活检组织中的淋巴细胞进行计数和表型分析。目标3是评估 使用系统生物学方法对直肠粘膜环境进行GRFT治疗， 利用现代质谱蛋白质组学分析来比较组织和直肠分泌物 从接受活性药物与安慰剂的动物，并评价GRFT治疗的效果 直肠微生物群的影响。