【摘要】 重组人生长激素（Recombinant human growth hormone, rhGH）是一种采用基因工程技术在大肠杆菌中表达的蛋白质药物，广泛用于治疗儿童及成人生长激素缺乏症等多种疾病。在rhGH药物工艺开发与生产过程中，可能产生多种产品相关物质，对药物质量构成潜在影响。因此，对rhGH主成分及产品相关物质进行深入的结构表征和合理的质量控制对保障药品质量具有重要意义。《欧洲药典》收录了用于rhGH产品相关物质质量控制的反相高效液相色谱方法，以及高丰度色谱峰的结构鉴定信息，但对于低丰度产品相关物质的结构分析并没有详细记载。本研究采用反相高效色谱与液相色谱-质谱联用技术，成功分离并鉴定了rhGH产品中的3种低丰度产品相关物质。研究结果表明，rhGH低丰度产品相关物质普遍含有多种修饰或罕见修饰，包括N99和N149双位点脱酰胺、N端缺失两个氨基酸同时N149脱酰胺、Q122脱酰胺、N端氨基甲酰化和N端丁二酰化，其中首次在rhGH中发现N端丁二酰化修饰。本研究对rhGH产品相关物质进行了深入表征，为rhGH药物的研发和工艺改进提供了依据。更多还原

【Abstract】 Recombinant human growth hormone(rhGH) is a protein drug expressed inEscherichia colivia genetic engineering, and plays a vital role in regulating growth, metabolism and energy balance. It is widely used in treatment of various clinical diseases, including growth hormone deficiencies in children and adults. During the development and production process of rhGH, various product-related substances may be formed, potentially impacting the drug′s quality. Therefore, detailed structural characterization and rigorous quality control of rhGH and its product-related substances are crucial for ensuring pharmaceutical quality. TheEuropean Pharmacopoeia includes methods for quality control of rhGH product-related substances using reversed-phase high-performance liquid chromatography(RP-HPLC) and provides structural identification information for high abundance chromatographic peaks. However, it lacks detailed records for the structural analysis of low abundance productrelated substances. In this study, RP-HPLC coupled with liquid chromatography-mass spectrometry(LC-MS) was used to successfully separate and identify three low abundance product-related substances of rhGH. The study revealed that low abundance product-related substances of rhGH generally contained multiple modifications or rare modifications, including double deamidation at N99and N149, simultaneous loss of two amino acids at the N-terminus and deamidation at N149, and deamidation at Q122, N-terminal formylation, and N-terminal butanoylation. Notably, the discovery of N-terminal butanoylation in rhGH was the first of its kind. This research provided a comprehensive and in-depth characterization of rhGH product-related substances, offering a reliable basis for the development and process improvement of rhGH drugs.更多还原