Manufacturing Scale-up, Purification, and Analytical Method Development, Validation and Qualification for KC1003, Key Precursor for ErythroMer (RBC substitute)

项目摘要/摘要 ErythroMer(EM)是一种新型的生物合成血液替代品，旨在解决严重未满足的血液需求 在无法使用储存的红血球(RBC)的情况下紧急输血或 不受欢迎。EM是一种自组装的脂质-多肽杂化纳米颗粒，每颗粒有效载荷高达 专门设计用于纠正以前的基于血红蛋白的氧气载体的故障的血红蛋白(Hb)， 这并不保留红细胞生理学的关键特征。受生物启发的EM设计超越了以前的 模仿红细胞特性的障碍：长期稳定，精确的动态反应变构效应器 控制Hb氧亲和力，控制Hb与一氧化氮(NO)的相互作用，防止血管痉挛，以及 通过有效载荷抗氧化剂减轻血红蛋白氧化。EM专为无菌冷冻干燥而设计 在环境条件下延长了保质期，并提供了规模化的成本效益生产。KaloCyte已经研发出 一个务实而有力的循序渐进的GMP商业化发展计划，包括规模- UP和cGMP制造红血球及其关键成分、定制的脂类前体和血红蛋白。 先前已完成的脂类前体的合成、放大和纯化的研究 从红血球(RBC)中提取的血红蛋白非常有前景。这些研究提供了强有力的证据 这一提议的前提和可行性，以及我们对制造规模、提纯 以及脂类前体的分析方法开发、验证和鉴定。Pre-Ind阶段 临床前研究要求EM数量大于CDMO服务所需的数量，并达到CDMO服务所需的质量。成功 完成本文件中提议的放大和分析工作将为KaloCyte提供 对KC1003的制造工艺以及用于制造的组件的信心 EM的质量是最高的，符合监管机构要求的可接受规格。 KC1003的制造放大将包括关键参数的特定规模优化 生产，如温度，时间，压力和反应物浓度在其每一步 制造过程。KC1003的分析方法开发、验证和优化应 按照FDA推荐的非物质文化遗产指南执行。方法验证数据的统计分析应 用来证明每种方法的有效性。完成产品开发过程中的这些步骤 商业化，将有助于在严格的GLP指南下完成EM的启用IND的临床前研究。埃姆 有能力在危及生命的失血情况下戏剧性地改变护理，这些情况下储存着红色血液 细胞(RBC)不可用(救护车、军事和严峻环境、不发达国家)， 不受欢迎的(输血风险超过益处的情况)，或供应不足(大规模伤亡事件)。

Project Summary/Abstract ErythroMer (EM) is a novel biosynthetic blood substitute developed to address the critically unmet need for emergency transfusion in situations where the use of banked red blood cells (RBCs) are inaccessible or undesirable. EM is a self-assembled lipid-peptidic hybrid nanoparticle with a high per particle payload of hemoglobin (Hb) that is specifically designed to rectify failures of previous hemoglobin-based oxygen carriers, which do not preserve key features of RBC physiology. The bio-inspired EM design surmounts previous obstacles by emulating RBC features: long term stability, precise dynamically-responsive allosteric effector control of Hb oxygen affinity, control of Hb interaction with nitric oxide (NO), preventing vasospasm, and mitigation of hemoglobin oxidation via payload antioxidants. EM is designed for sterile lyophilization enabling extended shelf life at ambient conditions and offers cost-effective production at scale. KaloCyte has developed a pragmatic yet robust stepwise goal-oriented development plan for GMP commercialization including the scale- up and cGMP manufacturing of ErythroMer and its key ingredients, customized lipid precursor, and hemoglobin. Previously completed studies on the synthesis and scale-up of the lipid precursor, and the purification of hemoglobin from red blood cells (RBCs) have been extremely promising. These studies provide strong evidence of the premise for and feasibility of this proposal and our commitment for the manufacturing scale-up, purification and analytical method development, validation, and qualification of the lipid precursor. The pre-IND stage preclinical studies require EM volumes larger than and at a quality that necessitates CDMO services. Successful completion of the scale-up and analytical work proposed in the current submission will provide KaloCyte the confidence in the manufacturing processes for KC1003, and that the components being used for the fabrication of EM are of the highest quality and fall within the acceptable specifications required by the regulatory agencies. The manufacturing scale-up of KC1003 will include scale specific optimization of key parameters for it manufacturing such as temperature, time, pressure, and reactant concentrations at each step of its manufacturing process. Analytical methods development, validation, and optimization for KC1003, shall be performed as per ICH guidelines recommended by the FDA. Statistical analysis of methods validation data shall be performed to demonstrate the validity of each method. Completion of these steps in the path to product commercialization, will help complete IND-enabling preclinical studies for EM under strict GLP guidelines. EM has the capability to dramatically transform care in life-threatening blood loss situations where stored red blood cells (RBCs) are unavailable (ambulances, military and austere environments, undeveloped countries), undesirable (situations in which risk of transfusion exceeds benefit), or in short supply (mass casualty events).