Institute of Pharmaceutical Analysis, College of Pharmacy, Jinan University, Guangzhou, 510632, China.
Laboratory of Analytical Pharmaceutical Chemistry, Department of Pharmaceutical Sciences, CIRM, University of Liege, CHU B36, B-4000, Liege, Belgium.
J Chromatogr A. 2021 Jun 21;1647:462147. doi: 10.1016/j.chroma.2021.462147. Epub 2021 Apr 15.
Drug-induced phospholipidosis (DIPLD) represents a big concern for both regulatory authorities and pharmaceutical companies in drug discovery. Many researches pointed out that the negatively charged intralysosomal lipids play an important role in the formation of DIPLD. To better mimic this negatively charged lipid surface, a novel immobilized artificial membrane (IAM) column was prepared via in situ copolymerization of 12-methacryloyl n-dodecylphosphocholine (MDPC) and 12-methacryloyl n-dodecylphosphoric acid (MDPA). By introducing MDPA, the surface of the resulting monolithic column can be maintained negatively charged over a broad pH range. Scanning electron microscopy, elemental analysis and nano-HPLC experiments were carried out to characterize the physicochemical properties and chromatographic performance of the obtained monolithic IAM column. The results of ζ-potential and retention mechanism studies indicate that both hydrophobic and electrostatic interactions contribute greatly to the retention of cation analytes owing to the existence of the negatively charged MDPA under acidic conditions. To better assess the DIPLD potency of drug, the molar ratio between MDPC and MDPA in the monolithic column was carefully optimized. The results show that the poly(MDPCPA-co-EDMA) column has the best predictability with only two false-positives (donepezil, flecainide) in qualitative analysis of 61 drugs.
药物诱导的磷脂蓄积症（DIPLD）是药物发现过程中监管机构和制药公司非常关注的问题。许多研究指出，带负电荷的溶酶体内脂质在 DIPLD 的形成中起着重要作用。为了更好地模拟这种带负电荷的脂质表面，通过原位共聚 12-甲基丙烯酰基-n-十二烷基磷酸胆碱（MDPC）和 12-甲基丙烯酰基-n-十二烷基磷酸（MDPA）制备了一种新型固定化人工膜（IAM）柱。通过引入 MDPA，所得整体柱的表面可以在较宽的 pH 范围内保持带负电荷。通过扫描电子显微镜、元素分析和纳升 HPLC 实验对所得整体 IAM 柱的理化性质和色谱性能进行了表征。ζ-电位和保留机制研究的结果表明，由于在酸性条件下存在带负电荷的 MDPA，疏水性和静电相互作用极大地促进了阳离子分析物的保留。为了更好地评估药物的 DIPLD 效力，仔细优化了整体柱中 MDPC 和 MDPA 的摩尔比。结果表明，聚（MDPCPA-co-EDMA）柱具有最佳的预测能力，在对 61 种药物进行定性分析时只有两种假阳性（多奈哌齐、氟卡尼）。
