Pre-Clinical Development of a Intranasal Bivalent gD2/gB2 HSV-2 Mucosal Vaccine

摘要 生殖器单纯疱疹病毒-2(HSV-2)感染是影响4000万人的主要公共卫生问题 在美国和全球5亿英镑。HSV-2感染的特点是疼痛复发，生殖器 疼痛，这可能与相当大的心理痛苦同时发生。传播可以在性行为中发生 在分娩期间接触和从母亲传给孩子，会产生致命的后果。尽管有几种疫苗已经被 显示出对病毒有很强的中和活性，目前还没有有效的疫苗来预防 单纯疱疹病毒2型。最近的研究表明，虽然全身、体液免疫是有益的，但共同诱导粘膜 豁免权是全面保护的先决条件。BlueWillow Biologics已经建立了NanoVax®，这是一种专有的油包油- 水纳米乳剂(NE01)佐剂技术，专为鼻腔免疫而设计。我们已经 证明了NE01能够增强抗原对免疫系统的靶向性，同时安全地引发 强大的体液、粘膜和细胞免疫反应，包括平衡诱导Th1、Th2和Th17 细胞。这一特征促进了有效疫苗的开发，这种疫苗可以诱导粘膜和细胞介导的 对包括HSV-2在内的各种呼吸道和性传播病原体提供全面保护的免疫力。 为了解决这一未得到满足的医疗需求，蓝柳生物公司正在开发一种新型的鼻腔双价疫苗 由NE01制定的两种HSV-2糖蛋白(GB2和GD2)组成。我们的临床前研究表明 由此产生的鼻腔疫苗(IN)GB2/GD2-NE01是一种有效的商业可行的预防药物 预防HSV-2感染的策略。我们已经优化了工艺，以便扩大生产 抗原，以实现高纯度GB2和GD2药物物质的可重复生产，规模足以 满足临床需求。重要的是，我们已经证明了药物的预防性等效性 从已建立的放大制造工艺准备到我们以前在实验室规模准备的那些。 这表明我们有能力制造大量的纯材料，同时保持完整性和 疫苗关键成分的活性。 在此IIB阶段SBIR计划中，BlueWillow Biologics将执行IN的剩余开发工作 GB2/GD2-NE01 HSV-2疫苗必须提交研究用新药(IND)申请。使用 根据第二阶段建立的已建立的放大流程，我们将完善GB2表达流程以改进 生产，生产两批每个抗原，连同NE01佐剂，并奉献材料 完成IND-Enabling GLP毒理学研究并制定稳定性临床材料 评估。这份材料的准备将使IND前与FDA的会议能够要求反馈 并批准我们的普洛斯毒理学研究设计。在完成普洛斯毒理学研究后，蓝柳将 准备好提交IND申请并启动第一阶段临床试验(不在此基金的覆盖范围内)。

ABSTRACT Genital herpes simplex virus-2 (HSV-2) infections are a major public health problem affecting >40 million people in the United States and >500 million globally. HSV-2 infection is characterized by painful recurring, genital sores and this can coincide with considerable psychological distress. Transmission can occur during sexual contact and from mother to child during birth with fatal consequences. Although several vaccines have been shown to elicit strong neutralizing activity against the virus, there is currently no effective vaccine to prevent HSV-2. Recent research has shown that while systemic, humoral immunity is beneficial, co-induction of mucosal immunity is prerequisite for full protection. BlueWillow Biologics has established NanoVax®, a proprietary oil-in- water nanoemulsion (NE01) adjuvant technology, that is designed for intranasal immunization. We've demonstrated the ability of NE01 to augment the targeting of antigens to the immune system while safely eliciting potent humoral, mucosal and cellular immune responses, including balanced induction of Th1, Th2 and Th17 cells. This feature facilitates the development of efficacious vaccines that induce mucosal and cell mediated immunity for full protection against various respiratory and sexually transmitted pathogens, including HSV-2. To address this unmet medical need, BlueWillow Biologics is developing a novel, intranasal bivalent vaccine comprised of two HSV-2 glycoproteins (gB2 and gD2) formulated in NE01. Our preclinical studies have shown that the resulting intranasal vaccine (IN) gB2/gD2-NE01 is an effective and commercially viable prophylactic strategy to prevent HSV-2 infections. We have optimized the processes for scale-up manufacturing of the antigens to achieve reproducible production of highly pure gB2 and gD2 drug substances at a scale sufficient to meet clinical demand. Importantly, we've demonstrated the prophylactic equivalency of the drug substances prepared from the established scale-up manufacturing processes to those we previously prepared at lab scale. This demonstrates our ability to manufacture high quantity of pure material while preserving the integrity and activities of the key vaccine components. In this Phase IIB SBIR program, BlueWillow Biologics will execute the remaining development work on the IN gB2/gD2-NE01 HSV-2 vaccine necessary to submit an investigational new drug (IND) application. Using the established scale-up processes established under Phase II, we will refine the gB2 expression process to improve yield, manufacture two batches of each antigen, along with the NE01 adjuvant, and dedicate the material towards completing an IND-enabling GLP toxicology study and formulating clinical material for stability assessment. The preparation of this material will enable a pre-IND meeting with the FDA to request feedback and approval of our GLP toxicology study design. Upon completion of the GLP toxicology study, BlueWillow will be ready to file an IND application and initiate a Phase 1 clinical trial (not covered under this funding).