School of Pharmacy, Xi'an Jiaotong University, 76# Yanta Westroad, Xi'an, Shaanxi, 710061, People's Republic of China.
Institute of Pharmaceutical Science and Technology, Western China Science &Technology Innovation Harbour, Xi'an, Shaanxi, 710115, People's Republic of China.
Anal Bioanal Chem. 2023 Mar;415(7):1371-1383. doi: 10.1007/s00216-023-04528-7. Epub 2023 Jan 18.
In this study, a novel cell membrane chromatography (CMC) model was developed to investigate cluster of differentiation 147 (CD147) targeted anti-tumor drug leads for specific screening and ligand-receptor interaction analysis by SNAP-tagged CD147 fusion protein conjugation and polystyrene microspheres (PS) modification. Traditional Chinese medicines (TCMs) are widely used in the treatment of cancer. CD147 plays important roles in tumor progression and acts as an attractive target for therapeutic intervention; therapeutic drugs for CD147-related cancers are limited to date. Thus, a screening method for active components in TCMs is crucial for the further research and development of CD147 antagonists. However, improvement is still needed to perform specific and accurate drug lead screening using the CMC-based method. Recently, our group developed a covalently immobilized receptor-SNAP-tag/CMC model using silica gel as carrier. Besides the carboxyl group on multi-step modified silica particles, the amino group of benzyl-guanine (BG, substrate of SNAP-tag) also possesses reactivity towards the carboxyl group on available carboxyl-modified PS. Herein, we used PS as carrier and an extended SNAP-tag with CD147 receptor to construct the PS-BG-CD147/CMC model for active compound investigation coupled with HPLC/MS and applied this coupled PS-BG-CD147/CMC-HPLC/MS two-dimensional system to drug lead screening from Nelumbinis Plumula extract (NPE) sample. In addition, to comprehensively verify the pharmacological effects of screened ingredients, a cell proliferation inhibition assay was performed, and the interaction between the ingredients and CD147 was studied by the frontal analysis method. This study developed a high-throughput PS-based CMC screening platform, which could be widely applied and utilized in chromatographic separation and drug lead discovery.
在这项研究中，开发了一种新型的细胞膜色谱（CMC）模型，通过 SNAP 标记的 CD147 融合蛋白缀合和聚苯乙烯微球（PS）修饰，用于特定的筛选和配体-受体相互作用分析，研究 CD147 靶向抗肿瘤药物先导物。传统中药（TCM）广泛用于癌症治疗。CD147 在肿瘤进展中起重要作用，是治疗干预的有吸引力的靶标；目前，针对 CD147 相关癌症的治疗药物有限。因此，筛选 TCM 中的活性成分对于 CD147 拮抗剂的进一步研究和开发至关重要。然而，使用基于 CMC 的方法进行特异性和准确的药物先导筛选仍需要改进。最近，我们小组开发了一种使用硅胶作为载体的共价固定化受体-SNAP 标记/CMC 模型。除了多步修饰硅胶颗粒上的羧基外，苯甲基鸟嘌呤（BG，SNAP 标记的底物）的氨基也对可用羧基修饰 PS 上的羧基具有反应性。在这里，我们使用 PS 作为载体和带有 CD147 受体的扩展 SNAP 标记构建 PS-BG-CD147/CMC 模型，用于与 HPLC/MS 相结合的活性化合物研究，并将这种偶联的 PS-BG-CD147/CMC-HPLC/MS 二维系统应用于从荷叶提取物（NPE）样品中进行药物先导筛选。此外，为了全面验证筛选成分的药理作用，进行了细胞增殖抑制测定，并通过前沿分析方法研究了成分与 CD147 之间的相互作用。本研究开发了一种高通量基于 PS 的 CMC 筛选平台，可广泛应用于色谱分离和药物先导物发现。
