Rapidly screening of pancreatic lipase inhibitors from Sibiraea angustata (rehd.) Hand.-Mazz. leaves using affinity ultrafiltration combined with HPLC-QTOFMS, molecular docking, targeted separation, and in vitro experimental verification

An integrated strategy was proposed for the rapid screening of pancreatic lipase inhibitors from Sibiraea angustata (Rehd.) Hand.-Mazz. (S. angustata) leaves based on affinity ultrafiltration, high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (HPLC-QTOFMS), molecular docking, targeted separation and in vitro experimental verification. A total of fifteen target compounds, including five flavonoids, one phenylpropanol glycoside and nine terpenoids, had been screened as pancreatic lipase inhibitors from the ethyl acetate and n-butanol fractions of S. angustata leaves. Among these target compounds, thirteen of them are reported as pancreatic lipase inhibitors for the first time. Hyperin, quercetin-3,4′-diglucoside and peltatoside showed stronger binding abilities with pancreatic lipase with ΔG values −9.9, −10.2 and −9.2 kcal/mol while the IC50 values were 0.987 ± 0.079, 0.718 ± 0.054 and 0.916 ± 0.069 mmol/L, respectively. The target compounds quercetin-3,4′-diglucoside and peltatoside were separated with purities higher than 98% using macroporous resin and preparative chromatography.本研究提出一种基于亲和超滤、高效液相色谱-四极杆飞行时间质谱（HPLC-QTOFMS）、分子对接、靶向分离及体外实验验证的整合策略，用于从窄叶鲜卑花（Sibiraea angustata (Rehd.) Hand.-Mazz.，简称S. angustata）叶片中快速筛选胰脂肪酶抑制剂。从窄叶鲜卑花叶片的乙酸乙酯萃取分部和正丁醇萃取分部中，共筛选得到15种胰脂肪酶抑制剂类目标化合物，其中包括5种黄酮类、1种苯丙醇苷类及9种萜类化合物。其中13种化合物为首次被报道具有胰脂肪酶抑制活性。金丝桃苷（Hyperin）、槲皮素-3,4′-二葡萄糖苷（quercetin-3,4′-diglucoside）及盾叶苷（peltatoside）与胰脂肪酶的结合能力更强，其吉布斯自由能变化（ΔG）值分别为-9.9、-10.2和-9.2 kcal/mol，半抑制浓度（IC50）分别为0.987 ± 0.079、0.718 ± 0.054及0.916 ± 0.069 mmol/L。本研究通过大孔树脂结合制备型色谱法，将目标化合物槲皮素-3,4′-二葡萄糖苷及盾叶苷进行分离纯化，其纯度均高于98%。