药效动力学研究中流式细胞术方法学验证概述

Overview of Validation of Flow Cytometry-based Methodology in Pharmacodynamic Study

药效动力学研究是药物开发过程中的重要组成部分,特别是有助于加快药物的临床前和早期临床研究。在抗肿瘤和免疫相关药物的药效动力学研究方面,基于流式细胞术的检测方法已被广泛用于细胞免疫表型和功能分析、受体占有率、蛋白表达和修饰、通路研究等方面。流式细胞术强大的分析能力和多参数的特点,使其成为药效动力学研究中十分重要的技术手段。流式细胞检测技术的方法性能在很大程度上决定了所获得的药效动力学数据的可靠程度和应用价值。然而,流式细胞术相对复杂和独特的技术特点,使方法的建立、验证与检测应用具有较大挑战。在目前药物研发的监管环境下,国内外都没有流式细胞术应用于药效动力学研究的指导原则出台。本文综述了流式细胞术在药效动力学研究中的应用领域、法规要求和相关技术指南、方法建立的重要关注点、方法学验证等几个方面的研究进展,以期为业内形成更多共识提供帮助。

Pharmacodynamics is an important part of the whole drug development process,and is especially helpful to accelerating preclinical development and early phase clinical study.Flow cytometry-based assays have been widely used in cellular immunotyping,functional analysis,receptor occupancy analysis,protein expression and modification analysis,and pathway analysis for evaluating the pharmacodynamics of biomarkers during the development of drugs for cancers and immune diseases.The analytical capabilities of flow cytometry as well as the characteristics of multi-parameters make it to be an important tool for pharmacodynamic studies.The assay performance largely determines the reliability and application value of the pharmacodynamic data.However,the unique and complicated technical characteristics of flow cytometry make the establishment,verification,detection and application challenging.In the current regulatory environment,there are no guidelines for the research of flow cytometry-based methodology applied in pharmacodynamics.In this paper,the application of flow cytometry in pharmacodynamics,regulatory requirements,related technical references,critical considerations in method development,and method validation are reviewed.