Formulation Research, Pharmaceutical Science and Technologies Unit, Pharmaceutical Profiling & Development Function, Eisai Co., Ltd., 1 Kawashimatakehaya-machi, Kakamigahara, Gifu 501-6195, Japan.
Formulation Research, Pharmaceutical Science and Technologies Unit, Pharmaceutical Profiling & Development Function, Eisai Co., Ltd., 1 Kawashimatakehaya-machi, Kakamigahara, Gifu 501-6195, Japan.
J Chromatogr B Analyt Technol Biomed Life Sci. 2024 Oct 15;1247:124317. doi: 10.1016/j.jchromb.2024.124317. Epub 2024 Sep 13.
Lipid nanoparticles (LNPs) are emerging nucleic acid delivery systems in the development of mRNA therapeutics such as the severe acute respiratory syndrome coronavirus 2 vaccines. However, a suitable analytical method for evaluating the encapsulation efficiency (EE) of the LNPs is required to ensure drug efficacy, as current analytical methods exhibit throughput issues and require long analysis times. Hence, we developed and validated an anion-exchange HPLC method using Analytical Quality by Design. Three critical method parameters (CMPs) were identified using risk assessment and Design of Experiments: column temperature, flow rate, and sodium perchlorate concentration. The CMPs were optimized using Face-Centered Central Composite Design. The discriminating power of the optimized HPLC method and RiboGreen assay was comparable. The main advantage of this method is that LNPs can be directly injected into the HPLC system without bursting the LNPs loaded with encapsulated poly(A). The optimized HPLC method was validated as robust, high-throughput, and sufficiently sensitive according to the ICH Q2 guidelines. We believe our findings could promote efficient LNPs-based drug development.
脂质纳米粒（LNPs）是新型核酸递送系统，在开发 mRNA 疗法（如 SARS-CoV-2 疫苗）中具有重要作用。然而，为了确保药物疗效，需要一种合适的分析方法来评估 LNPs 的包封效率（EE）。目前的分析方法存在通量问题，需要较长的分析时间。因此，我们采用分析质量设计（AQD）开发并验证了一种阴离子交换 HPLC 方法。通过风险评估和实验设计（DoE）确定了三个关键方法参数（CMP）：柱温、流速和高氯酸钠浓度。使用中心复合面设计（CCD）对 CMP 进行了优化。优化后的 HPLC 方法与 RiboGreen 测定法具有相当的区分能力。该方法的主要优点是可以直接将负载有包封 poly(A)的 LNPs 注入 HPLC 系统，而无需破坏 LNPs。根据 ICH Q2 指南，优化后的 HPLC 方法被验证为稳健、高通量和足够灵敏。我们相信我们的发现将促进高效的基于 LNPs 的药物开发。
