Tobacco-based Sustainable Squalene for Novel Vaccine Adjuvant Emulsions

项目摘要/摘要 在这项拨款申请中，SynShark(SBA认证小型企业)与北卡罗来纳州立大学合作， 将展示药用级角鲨烯的一种新的植物来源，作为一种天然佐剂。该团队将利用诺奇 稳定烟草角鲨烯产量的植物工程路线，并发现制造和分离角鲨烯的方法 用于快速应对大流行的分子。这项研究符合美国国家过敏症研究所和 传染病科(NIAID)，下设三个部门：艾滋病科(DAIDS)、过敏、免疫学和 移植(DAIT)和微生物学和传染病司(DMID)。具体的分支机构和计划 在这些部门下，有相关的需求说明：(I)疫苗研究方案(DAIDS)，(Ii)基础 免疫学处(DAIT)，(Iii)SBIR佐剂研究方案--细菌学和真菌学(DMID)。 这项活动的最终目标是成功地将一种快速衍生的自然资源商业化并造福于公众。 基于植物角鲨烯的佐剂，具有完全的生物相容性。佐剂在疫苗推广中起着关键作用 通过激发更强的免疫反应和减少有效成分的体积需求来发挥功效。被所有人 有证据表明，SARS-CoV-1免疫依赖于T细胞激活，疫苗应该解释 就为了这个。目前受监管的SARS-CoV-1疫苗使用的合成佐剂不能模拟强大的T细胞 应答，而天然角鲨烯已知能增强T细胞应答的幅度。但药用角鲨烯 来自鲨鱼的‘活体’，传达：1)相关污染物和病原体的风险，以及2)公众抵抗力。在……上面 另一方面，另一种天然佐剂Quillaja Saponaria油，花费了它的制造Fujifilm Disynth(在Novavax SARS-CoV-1疫苗)每公斤超过10万美元。这导致了Fujifilm DiSynth对这一应用的支持。 通过萜类代谢工程，SynShark已经成功地改变了 烟草中的碳分配途径允许增加三萜角鲨烯的积累。第一阶段 正在引入的任务与向市场交付非动物来源的商业化计划直接相关 天然佐剂，通过成熟的烟草制药技术在室内种植。第一阶段为 计划将这种创新和颠覆性的萜烯采购技术付诸实施，实现 SynShark在分子遗传学方面的研究进展。在第一阶段，烟草厂将被改造成研究 将构建物置于毛状体下直接在叶表面产生角鲨烯的可行性 促进剂，从而以最适合室内控制种植的方式稳定产量。在第二阶段，SynShark， 通过坚定的合作伙伴，将根据美国食品和药物管理局建立制造和提纯的可扩展性 管理指南。

Project Summary/Abstract In this grant application, SynShark (SBA Certified Small Business), in collaboration with North Carolina State University, will demonstrate a new plant source for pharmaceutical grade squalene as a natural adjuvant. The team will utilize novel plant-engineering routes to stabilize squalene yields in tobacco, and discover methods to manufacture and isolate the molecule for rapid pandemic response. This research matches the interests of the National Institute of Allergy and Infectious Diseases (NIAID) with three of its divisions: Division of Aids (DAIDS), Division of Allergy, Immunology and Transplantation (DAIT) and Division of Microbiology and Infectious Diseases (DMID). Specific branches and programs under these divisions with relevant statements of needs are: (i) the Vaccine Research Program (DAIDS), (ii) Basic Immunology Branch (DAIT), (iii) SBIR Adjuvant Research Program – Bacteriology and Mycology Branch (DMID). The ultimate goal of this activity is the successful commercialization and public benefit of a rapidly derived natural adjuvant based on plant squalene which is completely biocompatible. Adjuvants have a key role in promotion of vaccine efficacy through eliciting a stronger immune response and reducing the volumetric need of active ingredient. By all accounts, evidence was available that SARS-CoV-1 immunity relied on T-cell activation and that vaccines should account for that. Synthetic adjuvants being used by the current regulated SARS-CoV-1 vaccines are not simulating a strong T-cell response, while natural squalene known to enhance the magnitude of the T-cell response. But pharmaceutical squalene is sourced from shark ‘livering’, conveying: 1) the risk of associated pollutants and pathogens, and 2) public resistance. On the other hand, the other natural adjuvant Quillaja Saponaria oil, costs it’s manufacture Fujifilm Diosynth (in the Novavax SARS-CoV-1 vaccine) more than $100,000 per kilogram. This has led to Fujifilm Diosynth’s support of this application. Through terpenoid metabolic engineering, SynShark has already managed to successfully shift the fundamental carbon allocation pathways within tobacco allowing for increased accumulation of the triterpene squalene. The Phase I tasks being introduced are directly related to the commercialization plan to deliver to the market a non-animal sourced natural adjuvant which is grown indoors through proven tobacco pharmaceutical techniques. The Phase I period is planned to take this innovative and disruptive terpene sourcing technology to fruition by enabling the application of SynShark’s research advancements in molecular genetics. During Phase I, the tobacco plant will be engineered to study the feasibility of squalene production directly on the leaf surface by placing the constructs under trichome-specific promoters, thereby stabilizing yield in a manner best suited for controlled indoor growing. In the Phase II period, SynShark, through committed collaborators, will establish scalability of manufacturing and purification under US Food and Drug Administration guidelines.