Process development for manufacturing eCD4-Ig

eCD4-Ig 制造工艺开发

• 批准号: 10603836
• 负责人: MICHAEL DAVID ALPERT
• 金额: $ 30万
• 依托单位: EMMUNE, INC
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10603836
• 关键词: Affinity Chromatography; Anions; Antibodies; Applications Grants; Binding; Biological; Biological Assay; Buffers; Cell Line; Cells; Chromatography; Circulation; Clinic; Custom; Development; Enzyme-Linked Immunosorbent Assay; Exclusion; Formulation; Funding; Glycoengineering; Grant; HIV; HIV Envelope Protein gp120; HIV Infections; HIV Receptors; Half-Life; Hydrophobic Interactions; Immunologics; Injectable; Intravenous; Life Extension; Longevity; Marketing; Mass Spectrum Analysis; Measures; Monoclonal Antibodies; Mutation; Oral; Outsourcing; Patients; Perfusion; Pharmaceutical Preparations; Phase; Phase I Clinical Trials; Process; Production; Proteins; Proteolytic Processing; Regimen; Resistance; SIV; Site; Subcutaneous Injections; Sulfate; Technology Transfer; Temperature; Therapeutic; Time; Validation; Viscosity; Work; analytical method; antibody mimetics; improved; intravenous administration; intravenous injection; manufacture; manufacturing process development; manufacturing quality; manufacturing technology; mimetics; neonatal Fc receptor; pill; prevent; process optimization; receptor; sialylation; small molecule; small molecule therapeutics; subcutaneous; therapeutic protein; tyrosine O-sulfate

ABSTRACT eCD4-Ig is an antibody-like therapeutic for HIV that was constructed by fusing CD4 domains 1 and 2, and an HIV coreceptor-mimetic sulfopeptide, to the N- and C-termini of the constant Fc region of an antibody. eCD4-Ig will join an emerging class of long-acting therapeutics for treating and preventing HIV infection. As a consequence of being built from parts of HIV’s receptors, eCD4-Ig neutralizes 100% of HIV isolates, and even neutralizes simian immunodeficiency virus (SIV). Thus, eCD4-Ig presents fundamental barriers to escape. This grant will fund the work that is needed in between cell line development and IND-enabling studies to bring eCD4-Ig to the clinic. In Phase I, we will validate assays for critical quality attributes (CQAs) including the sulfotyrosine content of the coreceptor-mimetic sulfopeptide, and then show that the eCD4-Ig protein made by new cell lines meets pre-defined CQAs. In Phase II, we will optimize our upstream process, downstream process, formulation buffers, and generate a GLP lot of eCD4-Ig protein for IND-enabling studies. Features of the upstream process that will be optimized include media and feed selection, a temperature shift, and fed-batch versus perfusion culture. Our downstream process is based on convective chromatography, including for Protein A affinity chromatography, anion exchange (AEX) chromatography, and hydrophobic interaction (HIC) chromatography. These processes efficiently remove poorly sulfated eCD4-Ig, aggregates, and host cell protein (HCP). We will develop a formulation for intravenous injection, and a high-concentration formulation with acceptable viscosity for subcutaneous injection. Production of a GLP lot of protein for IND-enabling studies will be outsourced to a CMO after a technology transfer process. These are the essential next steps towards bringing eCD4-Ig to the clinic.

抽象的 eCD4-Ig 是一种 HIV 抗体样治疗药物，通过融合 CD4 结构域 1 和 2 构建而成，并且 HIV 辅助受体模拟磺肽，连接至抗体恒定 Fc 区的 N 端和 C 端。 eCD4-Ig 将加入一类新兴的长效疗法，用于治疗和预防 HIV 感染。 eCD4-Ig 由部分 HIV 受体构建而成，可中和 100% 的 HIV 分离株，并且 甚至可以中和猿猴免疫缺陷病毒 (SIV)，因此，eCD4-Ig 构成了根本性的障碍。 这笔赠款将资助细胞系开发和 IND 启动之间所需的工作。 将 eCD4-Ig 引入临床的研究在第一阶段，我们将验证关键质量属性 (CQA) 的检测。 包括辅助受体模拟磺肽的磺基酪氨酸含量，然后表明 eCD4-Ig 新细胞系产生的蛋白质符合预先定义的 CQA 在第二阶段，我们将优化我们的上游工艺， 下游工艺、配制缓冲液，并生成 GLP 批次的 eCD4-Ig 蛋白以用于 IND 启用 将优化的上游工艺的特征包括介质和进料选择、 温度变化以及分批补料培养与灌注培养的比较 我们的下游工艺基于对流。 色谱法，包括用于 Protein A 亲和色谱法、阴离子交换 (AEX) 色谱法、 和疏水相互作用 (HIC) 色谱法这些过程可有效去除硫酸化不良的物质。 eCD4-Ig、聚集体和宿主细胞蛋白 (HCP) 我们将开发一种用于静脉注射的制剂， 以及具有适合皮下注射的粘度的高浓度制剂。 用于 IND 研究的 GLP 批次蛋白质将在技术转让过程后外包给 CMO。 这些是将 eCD4-Ig 推向临床的重要后续步骤。