Russo Giacomo, Grumetto Lucia, Baert Mathijs, Lynen Frederic
Separation Science Group, Department of Organic and Macromolecular Chemistry, Ghent University, Krijgslaan 281, S4-bis, B-9000, Gent, Belgium; School of Applied Sciences, Sighthill Campus, Edinburgh Napier University, 9 Sighthill Ct, EH11 4BN, Edinburgh, United Kingdom.
Department of Pharmacy, School of Medicine and Surgery, University of Naples Federico II, Via D. Montesano, 49, I-80131, Naples, Italy; Consorzio Interuniversitario INBB, Viale Medaglie D'Oro, 305, I-00136, Rome, Italy.
Anal Chim Acta. 2021 Jan 15;1142:157-168. doi: 10.1016/j.aca.2020.11.003. Epub 2020 Nov 11.
A comprehensive two-dimensional liquid chromatography-based biomimetic platform (LCxLC) has been developed and validated for drug diffusion studies. Human serum albumin (HSA) and immobilized artificial membrane (IAM) were thereby used in the first (D) and second (D) separation dimension, respectively. While the former was meant to emulate the blood, the latter was instead intended to mimic the intestinal mucosa epithelium. Therefore, the experimental conditions, i.e. pH, temperature and buffer composition, were modulated to reflect faithfully in vivo conditions. 30 compounds, whose effective intestinal permeability (P) assayed in situ on humans by a validated technique was known from the literature, were used as model drugs. A good and orthogonal separation was achieved for the whole dataset, although for a better distribution of the most polar compounds in the elution window a segmented gradient elution program had to be employed. Interestingly, the passively uptaken compounds having the most favourable P populated a specific area of the 2D plots, implying that the affinity for HSA and IAM has to lie in specific ranges in order for a compound to be satisfactorily absorbed from the intestinal lumen. Although these results should be regarded as preliminary, this work paves an entirely new and unprecedented way to profile pharmaceutically relevant compounds for their in vivo absorption and distribution potential.
一种基于二维液相色谱的仿生平台（LCxLC）已被开发并验证用于药物扩散研究。在第一（D）和第二（D）分离维度中分别使用了人血清白蛋白（HSA）和固定化人工膜（IAM）。前者旨在模拟血液，而后者旨在模拟肠黏膜上皮。因此，对实验条件，即pH、温度和缓冲液组成进行了调节，以忠实地反映体内条件。30种化合物，其通过一种经过验证的技术在人体原位测定的有效肠通透性（P）从文献中可知，被用作模型药物。对于整个数据集实现了良好且正交的分离，尽管为了在洗脱窗口中更好地分布极性最强的化合物，必须采用分段梯度洗脱程序。有趣的是，具有最有利P值的被动摄取化合物聚集在二维图的特定区域，这意味着为了使化合物从肠腔中得到满意的吸收，其对HSA和IAM的亲和力必须处于特定范围内。尽管这些结果应被视为初步结果，但这项工作为分析药物相关化合物的体内吸收和分布潜力开辟了一条全新且前所未有的途径。
