A Unique Automated Bioreactor for Rearing Aseptic Mosquitoes from Larvae to Adults to Support Manufacture of Sanaria PfSPZ Products.

独特的自动化生物反应器，用于培育从幼虫到成虫的无菌蚊子，以支持 Sanaria PfSPZ 产品的生产。

• 批准号: 9620426
• 负责人: Peter F. Billingsley
• 金额: $ 30万
• 依托单位: SANARIA, INC.
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-18 至 2020-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9620426
• 关键词: Adult; Africa; African; Air; Anopheles Genus; Attenuated; Automation; Biological; Biological Assay; Bioreactors; Cessation of life; Characteristics; Chemoprophylaxis; Child; Clinical; Clinical Trials; Country; Culicidae; Custom; Cyclic GMP; Disease; Dissection; Ebola virus; Ensure; Environment; Epidemic; European; Excision; Falciparum Malaria; Geographic Locations; Germany; Grant; Growth; Hand; Harvest; Heating; Hour; Human; Human Resources; Immunity; Incubators; Individual; Infection; Infection prevention; Institutes; Larva; Licensing; Licensure; Malaria; Malaria Vaccines; Mali; Manuals; Mass Vaccinations; Medical; Methods; Nutrient; Output; Parasites; Phase; Phase III Clinical Trials; Plasmodium falciparum; Plasmodium falciparum vaccine; Prevention; Price; Process; Production; Pupa; Radiation; Reporting; Reproducibility; Risk; Safety; Salivary Glands; Sea; Shapes; Site; Small Business Innovation Research Grant; Sporozoite vaccine; Sporozoites; Stream; System; Tanzania; Testing; Tube; Vaccines; Waste Products; base; commercialization; cost; design; egg; fitness; flasks; human old age (65+); improved; malaria infection; manufacturing process; protective efficacy; scale up; seal; success; tool; transmission process

Project Summary/Abstract: The annual estimated burden of malaria has increased to 216M clinical episodes and 445,000 deaths, and annual losses of >$12B of GDP to malaria in Africa alone. The world needs a malaria vaccine. Sanaria’s Plasmodium falciparum sporozoite (PfSPZ) vaccines have protected 100% of subjects against controlled human malaria infection (CHMI) in five clinical trials in the U.S., Germany, Tanzania, and Mali; protected adults at least for 6 months against intense, heterogeneous transmission of Pf in two clinical trials in Mali, and are now being assessed in clinical trials in 5 months to 65-year old’s in 5 African countries and in adults in 2 European countries as well as 3 sites in the US. The plan is to initiate the first Phase 3 clinical trials to support submission of a Biologics License Application to the FDA in the US in the second half of 2018, and in Africa in the first half of 2019. Sanaria’s PfSPZ-based vaccines contain aseptic, purified, cryopreserved, live Plasmodium falciparum (Pf) sporozoites (SPZ) that are manufactured in aseptically reared Anopheles stephensi mosquitoes. The current method of aseptic mosquito production, while robust and reproducible, relies heavily upon manpower and large quantities of expensive consumables, and requires significant space. Semi-automation of mosquito dissection to remove salivary glands developed under another SBIR grant is expected to increase the efficiency of the process to 750-1,000 mosquitoes per hour per operator in the first half of 2018. Thus, production of mosquitoes is now the bottleneck and most expensive step in the GMP manufacture of PfSPZ vaccines. The overarching aim of this project is therefore to develop methods to reduce manpower and consumable costs and overall space requirements by developing a semi- automated system for culturing (manufacturing) aseptic mosquitoes. In specific aim 1, we will design, build and test a bioreactor for hands-free, aseptic culturing from eggs to larvae to adult mosquitoes. This bioreactor will be developed to 1) meet all GMP requirements, 2) remove the need for expensive consumables to increase output, 3) reduce human costs, 4) eliminate the need for many of the biological safety cabinets used in mosquito production, and 5) be adaptable for eventual removal of incubators from mosquito production. We will demonstrate that the new bioreactor is capable of maintaining the critical aseptic environment for the larval cultures, and that the mosquitoes raised in the larval bioreactor show equivalence with those produced by current methods. In specific aim 2 we will develop apparatus for automated harvesting of adult mosquitoes. Our aim is to be able to transfer newly emerged adult mosquitoes directly and hands free from the larval bioreactors into big aseptic mosquito containers in which the mosquitoes will be fed upon gametocytes and maintained until the day of extraction of PfSPZ from mosquitoes. Developing the larval bioreactor will lead to a sea change in Sanaria’s capacity to manufacture its PfSPZ products, putting Sanaria on track for mass-scale manufacturing at phase 3 GMP compliance at a significantly reduced cost.

项目摘要/摘要：每年预计疟疾负担已增至 2.16 亿临床病例 仅在非洲，就有 445,000 人死亡，每年 GDP 损失超过 12B 美元。 Sanaria 的恶性疟原虫子孢子 (PfSPZ) 疫苗可以保护 100% 的疟疾疫苗。 在美国、德国、美国进行的五项临床试验中，受试者针对受控人类疟疾感染 (CHMI) 坦桑尼亚和马里；保护成年人至少 6 个月免受 Pf 的强烈、异质传播 在马里进行了两项临床试验，目前正在 5 个非洲国家进行 5 个月内针对 65 岁老年人的临床试验评估 该计划是在 2 个欧洲国家以及美国的 3 个地点启动第一个项目。 支持向美国 FDA 提交生物制剂许可申请的 3 期临床试验 2018 年下半年在非洲上市，2019 年上半年在非洲上市。Sanaria 的基于 PfSPZ 的疫苗含有无菌、 纯化、冷冻保存的活恶性疟原虫 (Pf) 子孢子 (SPZ)，在 无菌饲养史氏按蚊目前的无菌生产方法，同时。 稳健且可重复，严重依赖人力和大量昂贵的耗材，并且 需要很大的空间来半自动化地解剖蚊子，以去除在其下发育的唾液腺。 另一项 SBIR 拨款预计可将流程效率提高至每小时 750-1,000 只蚊子 2018年上半年，运营商面临灭顶之灾。因此，蚊子的生产现在是瓶颈，也是最昂贵的 因此，该项目的首要目标是开发 PfSPZ 疫苗的 GMP 生产。 通过开发半成品来减少人力和消耗品成本以及整体空间需求的方法 培养（制造）无菌蚊子的自动化系统 在具体目标 1 中，我们将设计、建造。 并测试生物反应器，用于从卵到幼虫再到成蚊的免提无菌培养。 生物反应器将被开发为 1) 满足所有 GMP 要求，2) 消除对昂贵消耗品的需求 增加产量，3) 减少人力成本，4) 消除对许多生物安全柜的需求 用于蚊子生产，5) 适合最终去除蚊子孵化器 我们将证明新的生物反应器能够保持关键的无菌状态。 幼虫培养的环境，以及幼虫生物反应器中饲养的蚊子表现出等效性 在具体目标 2 中，我们将开发自动化设备。 我们的目标是能够直接转移新出现的成年蚊子。 将双手从幼虫生物反应器中解放出来放入大型无菌蚊子容器中，蚊子将在其中 以配子细胞为食并维持直至从蚊子中提取 PfSPZ 的那一天。 幼虫生物反应器将导致 Sanaria 生产 PfSPZ 产品的能力发生翻天覆地的变化， Sanaria 有望以显着降低的成本实现第三阶段 GMP 合规性大规模生产。