聚多巴胺修饰有序大-微孔MOF仿生矿化蛋白质新型复合材料用于HPLC手性药物分离研究

Chiral stationary phase in HPLC approach quickly dominates the field of chiral drug resolution, protein-based CSPs remain one of the most widely used CSPs. But their chromatographic columns have shortcomings to be solved in the aspect of low capacity, low efficiency, poor stability and the narrow separation“spectrum”. We have recently reported that MOFs with micro porous could effectively extract and adsorbe the small molecules, exclude of proteins. As a result, we construct highly oriented and ordered macro-pores within micro-pores metal-organic framework (MOF) single crystal materials called SOM-ZIF-8, adjusting aperture to load a series of proteins. Here we intend to adopt poly-dopamine modified SOM-ZIF-8 as the general matrix of CSPs, and to build a novel composite materials PDA@SOM-ZIF-8 by covalently immobilizing proteins, inspired by natural bio-mineralization processes. The afforded proteins-MOF serve as versatile and highly efficient chiral stationary phases towards various racemates in both normal and reverse phase of HPLC with low column backpressure, and demonstrating good column capacity, reusability and reproducibility due to the protection of MOF. Then, we try to adapt a combination strategy of the dynamics, thermodynamics and molecular docking method, to clarify the separation mechanism and law. At last, a novel micro-solid phase extraction coupled with HPLC would develop and validate for the pretreatment and the determination of enantiomers accurately and sensitively in practical pharmaceuticals and biological samples. Moreover, our study provides a better understanding of the quality control of chiral drugs, and the mechanisms of pharmacokinetic, pharmacology and toxicology, and is to provide valuable guidance on design of new drug separation system.

HPLC法中的手性固定相法在手性药物拆分领域占据主导，其中，蛋白质类是应用最广泛的CSPs。但其色谱柱容量低、柱效低、稳定性差、分离“谱”窄的问题亟待解决。我们前期发现微孔MOFs能够吸附小分子，排阻蛋白质。因此，我们定制有序大孔-微孔MOF单晶材料SOM-ZIF-8，调节孔径以负载不同类型的蛋白质。本项目拟采用多巴胺修饰的SOM-ZIF-8作为通用型CSPs基质，基于仿生矿化的策略构建PDA@SOM-ZIF-8共价键合蛋白质复合材料，将其作为HPLC固定相，在正、反相两种模式下实现对药物的“广”谱手性分离，提高柱容量、耐用性和分离效能，加速传质。并结合动力学、热力学和分子对接的手段，阐明分离机制和规律。采用微固相萃取结合HPLC检测，建立实际药品和生物样品中对映体高效灵敏的分析方法。这对于手性药物的质量控制、药动学，药理和毒理作用机制研究具有重要意义，并为构建手性药物分离新体系提供思路。

本项目的研究工作围绕功能导向MOFs及其复合材料的设计合成及其手性分离和环境方面应用的性能开展研究，致力于发展基于金属有机骨架材料及其蛋白质复合材料的手性选择性吸附剂、色谱填料和催化剂，取得了一系列研究成果。开展了MOFs共价键合蛋白质复合材料的可控制备及手性拆分的研究。我们探索复合材料在模型手性分子的对映选择性吸附和分离应用方面的研究，并探讨了对映选择性吸附和分离性能与MOFs复合蛋白质材料的构效关系。制备了基于MIL-101复合材料的液相色谱固定相，在正相和反相两种模式下实现对药物分子等有机混合物的“广谱”分离，提高了柱分离效能并加速传质。本项目通过动力学、热力学和分子对接的手段，阐明分离机制和规律及MOFs固定相与被分离物质之间的关系与相互作用。为MOFs材料在液相色谱领域的研究提供重要的理论依据和实验指导。此外，在自然基金的支持下，本项目围绕有序大孔和微孔MOF单晶材料ZIF-67在环境保护领域的应用开展了创新性研究，为去除对环境有害的有机和贵金属污染物的MOFs复合吸附剂和催化剂材料的研发开辟了新途径并提供了科学依据。