A Totally Synthetic Immunostimulator that Targets Toll-like Receptor 2 and NOD2: Toward Improved Influenza Vaccines

流感病毒引起季节性流行病和偶尔的大流行病，造成严重的全球发病率， mortality.新出现的流感感染的威胁刺激了疫苗的开发，以广泛诱导 保护和持久的免疫力。技术可以安全地提高免疫原性，超过 目前使用的佐剂对于推进疫苗设计和开发至关重要。虽然研究了几个 几十年来，已经确定了广泛的肽聚糖（PGN）免疫刺激特性，具有作为免疫调节剂的潜在应用。 佐剂、技术、监管和开发方面的挑战阻碍了天然PGN的发展， 疫苗应用。Teichos Laboratories，LLC（Teichos）专有技术生产一种平台， 通过化学酶促全合成真正的PGN核心结构的大分子免疫刺激生物制剂， 称为sPGN。原型sPGN TL-001的初步结果支持其作为疫苗佐剂的效用。TL-001 是一种单链的、未交联的大分子（ca. 20 - 200 kDa）sPGN，其由稳健的、可扩展的和 从市售小分子、标准试剂和单一酶进行灵活的全合成。 因此，TL-001与所有sPGN一样，不含外源性且通常为强效的污染物免疫反应性抗体， 与从微生物中分离的类似大分子相伴的材料。在实验室接种疫苗 在模型中，低剂量TL-001诱导通过TLR 2和NOD 2受体介导的强烈反应，以刺激 通过两个协同信号层协调先天性和适应性免疫反应， 抗体产生和T细胞活化。高剂量sPGN无毒性反应， 给实验室的动物服用。该提案寻求支持，以推进TL-001从发现 （初步数据）进入开发阶段（应用于疾病预防研究）， 原理实验以验证作为人流感疫苗佐剂的开发。我们将生产一批测试产品， TL-001，将被评估以确定TLR 2和其他细胞机制在介导细胞凋亡中的作用。 在人单核细胞和巨噬细胞中的免疫刺激应答。我们将表征抗体和T 在小鼠中使用流感H1亚单位疫苗接种模型观察细胞对TL-001的应答。预期的技术成功 在拟议的研究中，将有助于支持TL-001安全性的II期评估， 激发模型、替代给药途径和制剂开发。

Influenza viruses cause seasonal epidemics and occasional pandemics that inflict significant global morbidity and mortality. The threat of emerging influenza infections has stimulated development of vaccines to induce broadly protective and durable immunity. Technology that may boost immunogenicity safely beyond levels afforded by currently used adjuvants is critical to advancing vaccine design and development. While research over several decades has identified broad peptidoglycan (PGN) immunostimulatory properties with potential application as an adjuvant, technical, regulatory, and development challenges have prevented development of native PGN for vaccine applications. Teichos Laboratories, LLC (Teichos) proprietary technology produces a platform of macromolecular immunostimulatory biologics by chemoenzymatic total synthesis of authentic PGN core structure, termed sPGN. Preliminary results with a prototype sPGN, TL-001, support its utility as a vaccine adjuvant. TL-001 is a single-strand, uncrosslinked, macromolecular (ca. 20 – 200 kDa) sPGN produced by robust, scalable, and flexible total synthesis from commercially available small molecules, standard reagents, and a single enzyme. Therefore, TL-001, like all sPGN, is produced free of the adventitious, and often potent, contaminant immune reactive materials that accompany similar macromolecules isolated from microorganisms. In a laboratory vaccination model, low dose TL-001 induces a robust response that is mediated through TLR2 and NOD2 receptors to stimulate coordinated innate and adaptive immune responses through two synergistic layers of signalling that amplify antibody production and T cell activation. Neither toxicity nor reactogenicity were observed when high dose sPGN was administered to laboratory animals. This proposal seeks support to advance TL-001 from discovery (preliminary data) to development phase (application to disease prevention studies) by conducting proof-of- principle experiments to validate development as a human influenza vaccine adjuvant. We will produce a test lot of TL-001 that will be evaluated to define the role of TLR2 and other cellular mechanisms in mediating the immunostimulatory responses in human monocytes and macrophages. We will characterize the antibody and T cell responses to TL-001 using an influenza H1 subunit vaccination model in mice. Anticipated technical success in the proposed studies will help support Phase II assessments of TL-001 safety, protection in lethal influenza challenge models, alternate routes of administration, and formulation development.