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基于我们最近的免疫疗法 治疗慢性乙型肝炎(CHB)的专利病毒样水泡(VLV)平台。目标是 该项目的主要任务是完成CAPG-201的临床前研究和产品开发 疫苗。我们设计了这个载体来实现慢性乙肝患者的功能性治愈，其特征是 由于乙肝表面抗原(HBs Ag)的持续丢失(有或没有HBs Ag抗体 血清转换)，使用我们基于VLV的乙肝病毒(乙肝)疗法。我们已经开发出一种 能够诱导病毒特异性CD8+T细胞清除乙肝病毒感染的治疗性疫苗。AS 进度报告详述(见附载的报告)，我们已完成大部分阶段工作 II里程碑。使用VLV双重启动子和糖蛋白开关，我们增强了疗效， 正如在腺相关病毒(AAV)小鼠模型中所证明的那样。我们有 同时含有MHBs和HBcAg抗原的Carg-201可控制乙肝病毒 复制，小鼠血清中的乙肝表面抗原水平降低就是证据。血清浓度的降低 高抗原血症AAV-HBV80%以上小鼠的生物标记物水平具有高度显著性 提示Carg-201可打破免疫耐受，促进T细胞对乙肝病毒抗原的应答。AS 这是生产良好制造规范(GMP)级VLV的首次尝试，我们的任务是 这一点至关重要，需要创新的方法。美国食品和药物管理局(FDA) 在2019年4月提供了关于CAPG-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标/终点和我们的第一阶段研究设计的积极反馈， 包括给药方案和患者资格标准。因此，随着GMP的完成， 制造和动物毒理学研究，我们现在正在将Carg-201推向临床 审判(S)。为了完成FDA对IND申请所要求的研究，我们建议 第二阶段的以下具体目标：目标1.完全安全、非临床毒理学和 Carg-201的药代动力学研究目标2.进行嘉吉公司的产品开发研究- 201，包括制造工艺开发、放大可行性研究、提纯、 分析化验开发、配方和稳定性研究。目标3.启动GMP 临床级CAPG-201材料的制造和发布，并提交IND申请到 支持第一阶段评估。影响：该项目解决了未满足的紧急需求 CHB治疗性疫苗，利用之前由NIH SBIR第一和第二阶段资助的工作 格兰特。这一项目处于有利地位，能够取得持续而有力的进展，实现 一种治疗慢性乙型肝炎的有效免疫疗法的开发。

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.