背景:中药(TCM)具有广泛的应用,包括人类医疗保健相关的治疗和生物活性化合物的发现。然而,复杂的化学系统对基于化学材料的研究和质量控制提出了重大挑战。例如,板蓝根(BLG)颗粒是一种公认的中药制剂,广泛应用于临床治疗病毒感染。然而,其抗流感功效的化学基础尚不清楚.
目的:在本研究中,建立了基于生物活性和化学分析鉴定抗流感分子的系统发现策略,有助于板蓝根颗粒抗流感物质基础的分子阐明。
方法:用血凝素酶抑制(HAI)和神经氨酸酶抑制(NAI)试验比较不同组分BLG颗粒对H1N1、H5N1和H7N9病毒的抗流感活性。使用超高效液相色谱与四极轨道阱质谱联用(UHPLC-Q-ExactiveOrbitrapMS)对BLG颗粒及其级分中的化学成分进行了比较定性分析,其中使用了多质谱数据库平台和三种化合物鉴定策略。使用多种化学计量和数据比较策略分析了抗流感活性与化学成分特征之间的关联。
结果:结果表明,BLG颗粒的色谱级分F3和F4具有最高的抗流感活性。在BLG颗粒中共鉴定出88种化合物,包括31种生物碱,16种有机酸,10个核苷,8种苯丙素类化合物,6含硫化合物,5个氨基酸,4芳香化合物,3醛和酮,2类黄酮,1酒精,1碳水化合物,和1脂肪族化合物。在这些中,在级分F3-F4中鉴定了31个特征化合物作为具有抗流感活性的候选化合物。此外,首次在BLG颗粒及其原料板蓝根中鉴定出6-甲氧基喹啉和4-胍丁醛。
结论:在这项研究中,我们提出了一个系统的发现策略来彻底调查抗流感活性,化学鉴定,BLG颗粒的成分-活性关系。这些数据不仅为BLG颗粒活性的分子机制提供了更深入的理解,同时也为发现潜在的新型候选药物以及BLG颗粒的质量评价和控制提供了依据。
BACKGROUND: Traditional Chinese medicine (TCM) has a wide range of applications, including human healthcare-associated treatments and bioactive compound discovery. However, complex chemical systems present a significant challenge for chemical-material-based research and quality control. For instance, Banlangen (BLG) granules is a well-acknowledged TCM preparation widely used in clinical treatment of virus infection. However, its chemical basis of anti-influenza efficacy remains unclear.
OBJECTIVE: In the present study, a systematic discovery strategy for identifying anti-influenza molecules based on biological activities and chemical analysis was established to contribute to the molecular elucidation of the anti-influenza material basis of Banlangen granules.
METHODS: Hemagglutinase inhibition (HAI) and neuraminidase inhibition (NAI) assays were used to compare the anti-influenza activities of different fractions of BLG granules against H1N1, H5N1 and H7N9 viruses. A comparative qualitative analysis of the chemical constituents in BLG granules and their fractions was performed using ultra-high-performance liquid chromatography coupled with quadrupole orbitrap mass spectrometry (UHPLC-Q-Exactive Orbitrap MS), in which a multiple mass spectrometry database platform and three compound identification strategies were used. The association between anti-influenza activities and chemical constituent characteristics was analyzed using multiple stoichiometries and data comparison strategies.
RESULTS: The results showed that the chromatography fractions F3 and F4 of the BLG granules had the highest anti-influenza activity. A total of 88 compounds were identified in the BLG granules, including 31 alkaloids, 16 organic acids, 10 nucleosides, 8 phenylpropanoids, 6 sulfur-containing compounds, 5 amino acids, 4 aromatic compounds, 3 aldehydes and ketones, 2 flavonoids, 1 alcohol, 1 carbohydrate, and 1 aliphatic compound. Out of these, 31 characteristic compounds were identified in fractions F3-F4 as candidate compounds with anti-influenza activity. Additionally, 6-methoxyquinoline and 4-guanidinobutanal were identified in BLG granules and its raw material (Isatidis Radix) for the first time.
CONCLUSIONS: In this study, we proposed a systematic discovery strategy to thoroughly investigate the anti-influenza activity, chemical identification, and constituents-activity relationship of BLG granules. These data not only provided a deeper understanding of the molecular mechanism of the activity of BLG granules, but also presented a basis for the discovery of potential novel drug candidates and quality evaluation and control of BLG granules.