Zhuang Xiao-Cui, Zhang Yong-Li, Chen Gui-Lin, Liu Ye, Hu Xiao-Lan, Li Na, Wu Jian-Lin, Guo Ming-Quan
Key Laboratory of Plant Germplasm Enhancement and Specialty Agriculture, Wuhan Botanical Garden, Chinese Academy of Sciences, Wuhan 430074, China.
University of Chinese Academy of Sciences, Beijing 100049, China.
Pharmaceuticals (Basel). 2021 Apr 1;14(4):313. doi: 10.3390/ph14040313.
Previous reports have illustrated that the incidence and mortality of cancer are increasing year by year worldwide. In addition, the occurrence, development, recurrence and metastasis of cancer are closely related to inflammation, which is a kind of defensive response of human body to various stimuli. As an important medicinal plant in Africa, has been reported to have certain anti-inflammatory and anti-proliferative activities, but its specific components and mechanisms of action remain elusive. To tackle this challenge, affinity ultrafiltration with drug targets of interest coupled to high-performance liquid chromatography-mass spectrometry (AUF-HPLC-MS/MS) could be utilized to quickly screen out bioactive constituents as ligands against target enzymes from complex extracts of this plant. AUF-HPLC-MS/MS with four drug targets, i.e., cyclooxygenase-2 (COX-2), 5-lipoxygenase (5-LOX), topoisomerase I (Top I) and topoisomerase II (Top II) were used to rapidly screen and characterize the anti-inflammatory and anti-proliferative natural ligands from , and the resulting potential active compounds as ligands with specific binding affinity to COX-2, 5-LOX, Top I and Top II, were isolated with modern separation and purification techniques and identified with spectroscopic method like NMR, and then their antiinflammatory and anti-proliferative activities were tested to verify the screening results from AUF-HPLC-MS/MS. Compounds and , which screened out and identified from showed remarkable binding affinity to COX-2, 5-LOX, Top I and Top II with AUF-HPLC-MS/MS. In addition, 1 new compound (compound ), together with 5 known compounds were also isolated and identified from . The structure of compound was elucidated by extensive 1D, 2D NMR data and UPLC-QTOF-MS/MS. Furthermore, compounds and were further proved to possess both anti-inflammatory and anti-proliferative activities which are in good agreement with the screening results using AUF-HPLC-MS/MS. This work showcased an efficient method for quickly screening out bioactive components with anti-inflammatory and anti-proliferative activity from complex medicinal plant extracts using AUF-HPLC-MS/MS with target enzymes of interest, and also demonstrated that neolignanamides (compounds and ) from would be the active components responsible for its anti-inflammatory and anti-proliferative activity with the potential to treat cancer and inflammation.
先前的报道表明，全球范围内癌症的发病率和死亡率逐年上升。此外，癌症的发生、发展、复发和转移与炎症密切相关，炎症是人体对各种刺激的一种防御反应。作为非洲一种重要的药用植物，据报道具有一定的抗炎和抗增殖活性，但其具体成分和作用机制仍不清楚。为应对这一挑战，可利用与感兴趣的药物靶点相结合的亲和超滤-高效液相色谱-质谱联用技术（AUF-HPLC-MS/MS），从该植物的复杂提取物中快速筛选出作为针对靶酶配体的生物活性成分。采用具有四种药物靶点，即环氧合酶-2（COX-2）、5-脂氧合酶（5-LOX）、拓扑异构酶I（Top I）和拓扑异构酶II（Top II）的AUF-HPLC-MS/MS，快速筛选和鉴定该植物中的抗炎和抗增殖天然配体，并利用现代分离纯化技术分离出与COX-2、5-LOX、Top I和Top II具有特异性结合亲和力的潜在活性化合物作为配体，并用核磁共振等光谱方法进行鉴定，然后测试它们的抗炎和抗增殖活性，以验证AUF-HPLC-MS/MS的筛选结果。通过AUF-HPLC-MS/MS从该植物中筛选并鉴定出的化合物和，对COX-2、5-LOX、Top I和Top II表现出显著的结合亲和力。此外，还从该植物中分离并鉴定出1个新化合物（化合物）以及5个已知化合物。化合物的结构通过广泛的一维、二维核磁共振数据和超高效液相色谱-四极杆飞行时间质谱联用技术（UPLC-QTOF-MS/MS）得以阐明。此外，化合物和进一步被证明具有抗炎和抗增殖活性，这与使用AUF-HPLC-MS/MS的筛选结果高度一致。这项工作展示了一种利用带有感兴趣靶酶的AUF-HPLC-MS/MS从复杂药用植物提取物中快速筛选出具有抗炎和抗增殖活性的生物活性成分的有效方法，同时也证明了该植物中的新木脂酰胺类化合物（化合物和）将是其具有抗炎和抗增殖活性、有潜力治疗癌症和炎症的活性成分。
