High Throughput Assay to Identify New Drugs Against Vivax Malaria Liver Stages

高通量测定法鉴定抗间日疟原虫肝期新药

• 批准号: 8251982
• 负责人: STEPHEN Lev HOFFMAN
• 金额: $ 30万
• 依托单位: SANARIA, INC.
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-06-04 至 2014-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8251982
• 关键词: Antimalarials; Aotus primate; Biological Assay; Cell Line; Cells; Chemicals; Chemoprophylaxis; Coculture Techniques; Collaborations; Communities; Cultured Cells; Data; Development; Disease; Dose; Drug resistance; Equipment and supply inventories; Foundations; Genomics; Geographic Locations; Goals; Health; Hepatocyte; Human; In Vitro; International; Laboratories; Lead; Libraries; Liver; Malaria; Marketing; Methods; Monkeys; National Institute of Allergy and Infectious Disease; Parasites; Pattern; Pharmaceutical Preparations; Phase; Plasmodium falciparum; Plasmodium vivax; Preclinical Drug Evaluation; Publishing; Reading; Regimen; Relapse; Research; Screening procedure; Shipping; Ships; Small Business Innovation Research Grant; Sporozoites; Staging; System; United States National Institutes of Health; Vivax Malaria; Work; base; cell type; economic impact; high throughput screening; in vivo; induced pluripotent stem cell; meetings; mortality; novel strategies; prevent; resistant strain; success

DESCRIPTION (provided by applicant): Plasmodium vivax (Pv) is the 2nd most important human malaria parasite, causing more than 80 million cases annually, and significant severe disease and mortality that were previously unrecognized. Recent data indicate that the impact of Pv malaria on health and economies of the developing world has been dramatically underestimated. Pv is responsible for at least 30% of malaria in U.S. travelers. Efforts to prevent and control Pv malaria have had less than expected success due to emergence of drug resistant strains and repeated relapses from dormant liver stage called the hypnozoite. The international malaria research community has recently emphasized the critical need for increased research on Pv, if there is be successful elimination of this parasite. Modern approaches to development of new drugs against Pv, including hypnozoites are urgently needed. Sanaria is the only laboratory in the world, which has a functional assay in place for studying the effects of drugs against Pv liver stages and inventory of Pv sporozoites (PvSPZ) that can be used in such an assay. Many drug libraries are available for screening, but cannot be screened against Pv, because of lack of a medium or high throughput assay. The goal of this Phase I SBIR is to build on our current assay and inventory of PvSPZ to develop such an assay, initiate the screen for drugs against the liver stages of Pv, including hypnozoites. The long-term goal is to use the system to identify and develop new drugs that will eliminate all Pv liver stage parasites, including hypnozoites. Such drugs will be ideal for chemoprophylaxis of malaria in travelers and for mass-administration to eliminate Pv from defined geographic areas, therefore having both developed- and developing-world markets estimated to exceed $1 billion annually. Specifically we will: 1) Develop a reproducible, consistent, robust system for culturing liver stages of Pv and use it to screen for drugs against the liver stages of Pv. This will include assessing the use of HepG2 cells, primary human hepatocytes (PHH), and induced pluripotent stem cell-derived human hepatocytes (iPSDH) to establish an optimal cell- based screening system. 2) In collaboration with the Bhatia lab at MIT develop a reproducible, consistent, robust system of culturing Pv liver stages for at least 3-6 wks to identify and enrich for hypnozoites and screen drugs for activity against Pv hypnozoites. A micro-patterned co-culture system using PHH or iPSDH is being optimized to attain this goal. 3) In collaboration with the Inglese group at the NIH Chemical Genomics Center (NCGC) use the methods developed in Specific Aims 1 and 2 to screen a 2,500 compound library to identify target compounds with activity against liver stage parasites, including hypnozoites. 4) In collaboration with the Wellems group at NIAID/NIH produce, purify, and cryopreserve at least 2x108 fully infectious PvSPZ. In 2010, in collaboration with the Wellems group, we produced 108 fully infectious PvSPZ. These additional PvSPZ will be used in Phase II to conduct full scale screening of large compound libraries to identify and then characterize and develop lead compounds for development as anti-liver stage drugs. 1 PUBLIC HEALTH RELEVANCE: Plasmodium vivax (Pv) is the 2nd most important human malaria parasite, causing more than 80 million cases annually, and significant severe disease, mortality, and economic impact that were previously unrecognized. Modern approaches to development of new drugs against Pv are urgently needed. The goal of this project is develop an assay system to identify and develop new drugs that will be ideal for chemoprophylaxis of malaria in travelers and for mass-administration to eliminate Pv from a geographic area.

描述（由申请人提供）：间日疟原虫 (Pv) 是第二重要的人类疟疾寄生虫，每年导致超过 8000 万例病例，以及以前未被认识到的严重疾病和死亡。最近的数据表明，疟疾对发展中国家健康和经济的影响被大大低估了。美国旅行者中至少 30% 的疟疾是由光伏引起的。努力预防 由于耐药菌株的出现以及从称为催眠体的休眠肝脏阶段反复复发，控制疟疾的成功率低于预期。国际疟疾研究界最近强调，如果要成功消除这种寄生虫，就迫切需要加强对疟疾的研究。迫切需要现代方法来开发针对 Pv 的新药物，包括催眠孢子。 Sanaria 是世界上唯一拥有功能测定法的实验室，用于研究药物对 Pv 肝阶段的影响以及可用于此类测定法的 Pv 子孢子 (PvSPZ) 清单。许多药物库可用于筛选，但由于缺乏中通量或高通量测定，无法针对 Pv 进行筛选。第一阶段 SBIR 的目标是在我们目前的 PvSPZ 检测和库存的基础上开发这样的检测方法，启动针对 Pv 肝脏阶段（包括催眠子）的药物筛选。长期目标是利用该系统识别和开发新药，消除所有 Pv 肝期寄生虫，包括休眠子。此类药物对于旅行者疟疾的化学预防和大规模给药以消除特定地理区域的疟疾来说是理想的选择，因此发达国家和发展中国家的市场每年估计超过 10 亿美元。具体来说，我们将：1）开发一个可重复的、一致的、强大的系统来培养 Pv 的肝脏阶段，并用它来筛选针对 Pv 的肝脏阶段的药物。这将包括评估 HepG2 细胞、原代人肝细胞 (PHH) 和诱导多能干细胞衍生的人肝细胞 (iPSDH) 的使用，以建立最佳的基于细胞的筛选系统。 2) 与麻省理工学院的 Bhatia 实验室合作，开发一个可重复、一致、稳健的 Pv 肝脏阶段培养系统，培养至少 3-6 周，以识别和富集催眠子和 筛选针对 Pv 休眠子活性的药物。为了实现这一目标，正在优化使用 PHH 或 iPSDH 的微图案共培养系统。 3) 与 NIH 化学基因组中心 (NCGC) 的 Inglese 小组合作，使用特定目标 1 和 2 中开发的方法筛选 2,500 种化合物库，以鉴定具有抗肝期寄生虫（包括催眠虫）活性的目标化合物。 4) 与 NIAID/NIH 的 Wellems 小组合作，生产、纯化和冷冻保存至少 2x108 完全感染性的 PvSPZ。 2010 年，我们与 Wellems 团队合作，生产了 108 个完全感染性的 PvSPZ。这些额外的 PvSPZ 将在第二阶段用于对大型化合物库进行全面筛选，以识别、表征和开发先导化合物，以开发为抗肝阶段药物。 1 公共卫生相关性：间日疟原虫 (Pv) 是第二重要的人类疟原虫，每年造成超过 8000 万例病例，并造成以前未被认识到的严重疾病、死亡率和经济影响。迫切需要现代方法来开发抗PV新药。该项目的目标是开发一种检测系统，以识别和开发新药，这些新药对于旅行者疟疾的化学预防和大规模给药以消除某个地理区域的疟疾来说是理想的选择。