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评估生物类似药的分析可比性:以粒细胞集落刺激因子(GCSF)为例

Assessing analytical comparability of biosimilars: GCSF as a case study.

作者信息

Nupur Neh, Singh Sumit Kumar, Narula Gunjan, Rathore Anurag S

机构信息

Department of Chemical Engineering, IIT Delhi, Hauz Khas, New Delhi 110016, India.

Department of Chemical Engineering, IIT Delhi, Hauz Khas, New Delhi 110016, India.

出版信息

J Chromatogr B Analyt Technol Biomed Life Sci. 2016 Oct 1;1032:165-171. doi: 10.1016/j.jchromb.2016.05.027. Epub 2016 May 21.

Abstract

The biosimilar industry is witnessing an unprecedented growth with the newer therapeutics increasing in complexity over time. A key step towards development of a biosimilar is to establish analytical comparability with the innovator product, which would otherwise affect the safety/efficacy profile of the product. Choosing appropriate analytical tools that can fulfil this objective by qualitatively and/or quantitatively assessing the critical quality attributes (CQAs) of the product is highly critical for establishing equivalence. These CQAs cover the primary and higher order structures of the product, product related variants and impurities, as well as process related impurities, and host cell related impurities. In the present work, we use such an analytical platform for assessing comparability of five approved Granulocyte Colony Stimulating Factor (GCSF) biosimilars (Emgrast, Lupifil, Colstim, Neukine and Grafeel) to the innovator product, Neupogen(®). The comparability studies involve assessing structural homogeneity, identity, secondary structure, and product related modifications. Physicochemical analytical tools include peptide mapping with mass determination, circular dichroism (CD) spectroscopy, reverse phase chromatography (RPC) and size exclusion chromatography (SEC) have been used in this exercise. Bioactivity assessment include comparison of relative potency through in vitro cell proliferation assays. The results from extensive analytical examination offer robust evidence of structural and biological similarity of the products under consideration with the pertinent innovator product. For the most part, the biosimilar drugs were found to be comparable to the innovator drug anomaly that was identified was that three of the biosimilars had a typical variant which was reported as an oxidized species in the literature. But, upon further investigation using RPC-FLD and ESI-MS we found that this is likely a conformational variant of the biotherapeutic been studied.

摘要

随着新型治疗药物的复杂性与日俱增,生物类似药行业正经历前所未有的增长。生物类似药研发的关键一步是与创新产品建立分析可比性,否则会影响产品的安全性/有效性。选择能够通过定性和/或定量评估产品关键质量属性(CQAs)来实现这一目标的合适分析工具,对于确立等效性至关重要。这些CQAs涵盖产品的一级和高级结构、与产品相关的变体和杂质,以及与工艺相关的杂质和宿主细胞相关的杂质。在本研究中,我们使用这样一个分析平台来评估五种已批准的粒细胞集落刺激因子(GCSF)生物类似药(Emgrast、Lupifil、Colstim、Neukine和Grafeel)与创新产品Neupogen(®)的可比性。可比性研究包括评估结构均一性、同一性、二级结构以及与产品相关的修饰。本研究使用了物理化学分析工具,包括带质量测定的肽图分析、圆二色(CD)光谱、反相色谱(RPC)和尺寸排阻色谱(SEC)。生物活性评估包括通过体外细胞增殖试验比较相对效价。广泛分析检测的结果提供了有力证据,证明所研究的产品与相关创新产品在结构和生物学上具有相似性。在很大程度上,发现生物类似药与创新药具有可比性,所发现的异常情况是,三种生物类似药有一种典型变体,文献报道该变体为氧化形式。但是,通过RPC-FLD和ESI-MS进一步研究发现,这可能是所研究生物治疗药物的一种构象变体。

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