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

ErythroMer（红细胞替代品）的关键前体 KC1003 的生产放大、纯化和分析方法开发、验证和鉴定

• 批准号: 10324723
• 负责人: Esma Alp
• 金额: $ 50万
• 依托单位: KALOCYTE, INC.
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-05-11 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10324723
• 关键词: Address; Affinity; Ambulances; Antioxidants; Area; Biological Assay; Biological Availability; Blood; Blood Substitutes; Blood Vessels; Caring; Cell physiology; Classification; Complex; Country; Custom; Cyclic GMP; Data; Detection; Development; Development Plans; Emergency Situation; Endotoxins; Engineering; Environment; Erythrocytes; Esterification; Evaluation; Event; Failure; Formulation; Freeze Drying; Goals; Guidelines; Hemoglobin; Hemorrhage; High Pressure Liquid Chromatography; Human; Hybrids; Hydrogenation; Hydrolysis; Laboratories; Life; Lipids; Methodology; Methods; Military Personnel; Modeling; Morphology; Multiple Trauma; Nitric Oxide; Nucleosome Core Particle; Oxygen; Patient Care; Peptides; Pharmacologic Substance; Pharmacopoeias; Preparation; Procedures; Process; Production; Reaction Time; Readiness; Reference Standards; Reflex action; Risk; Sampling; Series; Services; Shapes; Specific qualifier value; Statistical Data Interpretation; Sterility; Structure; System; Temperature; Testing; Text; Time; Transfusion; United States; Validation; Vasospasm; Weight; Work; advanced analytics; amphiphilicity; analytical method; base; biomaterial compatibility; commercialization; cost effective; design; detection limit; detection sensitivity; experience; falls; innovation; interest; manufacturing process; manufacturing scale-up; mass casualty; method development; microbial; nano; nanoparticle; novel; oxidation; particle; point of care; portability; pre-clinical; preclinical safety; preclinical study; pressure; prevent; reconstitution; scale up

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).

项目总结/摘要 ErythroMer（EM）是一种新型的生物合成血液替代品，旨在解决以下严重未满足的需求： 在无法使用库存红细胞（RBC）的情况下进行紧急输血，或 不受欢迎。EM是一种自组装的脂质-肽混合纳米颗粒，具有高的每颗粒有效载荷， 血红蛋白（Hb），其专门设计用于纠正先前基于血红蛋白的氧载体的失效， 其不保留RBC生理学的关键特征。生物灵感的EM设计超越了以前的 通过模拟RBC特征来克服障碍：长期稳定性，精确的动态响应变构效应物 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，和 通过有效载荷抗氧化剂减轻血红蛋白氧化。EM设计用于无菌冻干， 延长了环境条件下的保质期，并提供了具有成本效益的大规模生产。KaloCyte开发了 一个务实而稳健的逐步目标导向的GMP商业化发展计划，包括规模- 生产ErythroMer及其关键成分、定制脂质前体和血红蛋白。 先前完成的关于脂质前体的合成和规模扩大以及脂质前体的纯化的研究， 来自红细胞（RBC）的血红蛋白是非常有前途的。这些研究提供了强有力的证据 该方案的前提和可行性，以及我们对生产规模扩大、纯化 以及脂质前体的分析方法开发、验证和确认。Pre-IND阶段 临床前研究需要的EM体积大于CDMO服务所需的质量。成功 完成当前提交文件中提出的扩大规模和分析工作将为KaloCyte提供 对KC 1003的制造过程有信心，并且用于制造的组件 的EM具有最高质量，并符合监管机构要求的可接受规格。 KC 1003的生产规模扩大将包括其关键参数的规模特定优化 制造过程中的每一步的温度、时间、压力和反应物浓度等因素， 制造过程KC 1003的分析方法开发、验证和优化应 按照FDA推荐的ICH指南进行。方法验证数据的统计分析应 以证明每种方法的有效性。在产品开发过程中完成这些步骤 商业化，将有助于在严格的GLP指导原则下完成EM的IND使能临床前研究。EM 有能力极大地改变危及生命的失血情况下的护理， 细胞（红细胞）不可用（救护车，军事和严峻的环境，不发达国家）， 不受欢迎（输血风险超过益处的情况）或供应短缺（大规模伤亡事件）。