GMP MANUFACTURE OF CLINICAL GRADE THERAPEUTIC VACCINE FOR THE TREATMENT OF PATIENTS WITH CHRONIC HBV

这个SBIR项目的最终目标是开发一种免疫疗法， 获得专利的病毒样囊泡（VLV）平台，用于治疗慢性B型肝炎（CH B）。目标 该项目的目的是完成CARG-201的临床前研究和产品开发 疫苗我们设计了这种载体，以实现慢性乙型肝炎患者的功能性治愈，其特征在于 由于持续丧失B型肝炎表面抗原（HBsAg）（有或无HBsAg抗体 血清转化），使用我们的基于VLV的B型肝炎病毒（HBV）疗法。我们已经开发出一种 能够诱导清除HBV感染的病毒特异性CD 8 + T细胞的治疗性疫苗。作为 根据进度报告（见附件RPRR）中的详细说明，我们已经完成了本阶段的大部分工作， 二.里程碑。使用VLV双启动子和糖蛋白开关，我们增强了功效， 如在持续性HBV的腺相关病毒（AAV）小鼠模型中所证实的。我们有 产生了携带MHBs和HBcAg抗原的CARG-201，其可以控制HBV 复制，如小鼠血清HBsAg水平降低所证明。血清中的减少 具有高抗原血症的超过80%的AAV-HBV小鼠中的生物标志物水平是高度显著的， 这表明CARG-201可以打破耐受性并驱动T细胞对HBV抗原的应答。作为 这是首次尝试生产符合良好生产规范（GMP）的超大型运载火箭，我们的任务是 这是非常重要的，需要创新的方法。美国食品药品监督管理局（FDA） 于2019年4月提供了关于CARG-201的研究前新药（IND）反馈，我们已经 解决了他们所有的担忧。FDA同意了我们提出的动物毒理学计划 程序、GMP生产方法和质量控制测试。FDA还提供了 对我们的临床研究目的/终点和1期研究设计的积极反馈， 包括给药方案和患者资格标准。随着GMP的完成， 生产和动物毒理学研究，我们现在正在进行CARG-201的临床 审判。为了完成FDA要求的IND申请研究，我们建议 第二阶段b的具体目标如下：目标1。完整的安全性、非临床毒理学和 CARG-201的药代动力学研究。目标2.进行CARG的产品开发研究- 201，包括制造工艺开发、规模扩大可行性研究、纯化、 分析测定开发以及制剂和稳定性研究。目标3：启动GMP 临床级CARG-201材料的生产和放行，并向 支持第一阶段评估。影响：该项目解决了未得到满足的紧急需求， CHB治疗性疫苗，利用先前由NIH SBIR I期和II期资助的工作 格兰特.该项目完全有能力取得持续和有力的进展， 开发有效的CHB免疫疗法。

The ultimate objective of this SBIR project is to develop an immunotherapy based on our recently patented virus-like vesicle (VLV) platform for the treatment of chronic hepatitis B (CHB). The goal of this project is to complete preclinical studies and product development of the CARG-201 vaccine. We have designed this vector to achieve a functional cure in CHB patients, characterized by a sustained loss of hepatitis B surface antigen (HBsAg) (with or without HBsAg antibody seroconversion), using our VLV-based hepatitis B virus (HBV) therapies. We have developed a therapeutic vaccine capable of inducing virus-specific CD8+ T cells that clear HBV infection. As detailed in the progress report (see attached RPRR), we have attained the majority of the Phase II milestones. Using a VLV dual promoter and a glycoprotein switch, we have enhanced efficacy, as demonstrated in an adeno-associated virus (AAV) mouse model of persistent HBV. We have generated CARG-201 harboring both MHBs and HBcAg antigens, which can control HBV replication, as evidenced by reduced serum HBsAg levels in mice. The reduction in serum biomarker levels in more than 80% AAV-HBV mice with high antigenemia is highly significant as it indicates that CARG-201 can break tolerance and drive T-cell responses to HBV antigens. As this is the first attempt to produce good manufacturing practice (GMP)-grade VLVs, our task is critically important and requires innovative approaches. The Food and Drug Administration (FDA) provided pre-investigational new drug (IND) feedback on CARG-201 in April 2019, and we have addressed all of their concerns. The FDA agreed with our proposed plan on the animal toxicology program, GMP manufacturing methods, and quality control testing. The FDA also provided positive feedback on our clinical study objectives/endpoints and our Phase 1 study design, including the dosing regimen and patient eligibility criteria. Thus, with the completion of GMP manufacturing and animal toxicology studies, we are now progressing CARG-201 to clinical trial(s). To complete the studies required by the FDA for an IND application, we propose the following specific aims in Phase IIb: Aim 1. Complete safety, non-clinical toxicology, and pharmacokinetics studies of CARG-201. Aim 2. Conduct product development studies of CARG- 201, including manufacturing process development, scale-up feasibility studies, purification, analytic assay development, and formulation and stability studies. Aim 3. Initiate GMP manufacture and release of clinical-grade CARG-201 material and submit an IND application to support Phase I evaluation. IMPACT: This project addresses the urgent unmet need for a therapeutic vaccine for CHB, harnessing work previously funded by an NIH SBIR Phase I and II grant. This project is well positioned to yield sustained and powerful progress towards the development of an efficacious immunotherapy for CHB.