Abstract：
The chemical composition of Ganoderma lucidum ethanol extracts was systematically analyzed and identified by ultra-high performance liquid chromatography-quadrupole electrostatic field orbitrap high-resolution mass spectrometry(UPLC-Orbitrap-HRMS). The fragmentation pattern of the representative chemical compounds was summarized, and the potential anti-liver fibrosis active compounds of G. lucidum acting on the farnesoid X receptor(FXR) target were studied to elucidate its pharmacodynamic substance basis. Preliminarily, 95 chemical constituents of G. lucidum ethanol extracts were identified, including 24 ganoderic acids, 9 ganoderenic acids, 13 lucidenic acids, 3 ganolucidic acids, 1 ganoderma lactone, 40 other triterpenoids, 4 fatty acids, and 1 other constituent. In addition, the fragmentation patterns of the representative compounds were also analyzed. The structural characteristics of ganoderic acids and ganoderenic acids were the C30 skeleton, containing free-COOH and-OH groups, which could easily lose H_2O and CO_2 to form fragment ions. The D-ring was mostly a five-membered ring, which was prone to breakage. Lucidenic acids were the lanosterolane-type of the C27 skeleton, and the side-chain structure became shorter and contained the same free-COOH and-OH compared with ganoderic acids, which had been reduced from 8 to 5 cartons and prone to lose H_2O and CO_2. Then, six reported FXR receptor agonists were selected to form a training set for establishing a pharmacophore model based on FXR ligands. The 95 identified chemical constituents of G. lucidum were matched with the pharmacophore, and the optimal pharmacophore model 02(sensitivity=0.750 00, specificity=0.555 56, ROC=0.750) was selected for the virtual screening of the G. lucidum compound library through the validation of the test set. Finally, 31 potential G. lucidum active constituents were screened and chosen to activate the FXRs. The ADMET results showed that ganoderic acid H and lucidenic acid J had less than 90% plasma protein binding rate and no hepatotoxicity, which could be used as FXR activators for developing clinical drugs for the treatment of liver fibrosis, either alone or in combination.
摘要：
采用超高效液相色谱-四极静电场轨道阱高分辨质谱(UPLC-Orbitrap-HRMS)对灵芝乙醇提取物的化学成分进行了系统分析和鉴定。总结了代表性化合物的碎裂模式，并对灵芝的潜在抗肝纤维化活性化合物作用于法尼醇X受体（FXR）靶点进行了研究，以阐明其药效学物质基础。初步而言,鉴定了灵芝乙醇提取物的95个化学成分，包括24种灵芝酸,9根肾上腺素酸，13种Lucidenicacids,3甲戊酸，1个灵芝内酯,40种其他三萜类化合物，4脂肪酸，和1个其他成分。此外,还分析了代表性化合物的断裂模式。灵芝酸和甘戊酸的结构特征为C30骨架，含有游离-COOH和-OH基团，容易失去H_2O和CO_2形成碎片离子。D环主要是五元环，容易破损。Lucidenicacids是C27骨架的羊毛甾醇型，与灵芝酸相比，侧链结构变得更短，含有相同的游离-COOH和-OH，从8箱减少到5箱，容易损失H_2O和CO_2。然后，我们选择了6种报道的FXR受体激动剂,以形成基于FXR配体建立药效团模型的训练集.鉴定出的95个灵芝化学成分与药效团相匹配,通过对测试集的验证，选择最佳药效团模型02（灵敏度=0.75000，特异性=0.55556，ROC=0.750）用于灵芝化合物文库的虚拟筛选。最后,筛选并选择31种潜在的灵芝活性成分以激活FXR。ADMET结果显示,灵芝酸H和葡萄糖酸J血浆蛋白结合率低于90%,无肝脏毒性,可用作FXR激活剂，用于开发治疗肝纤维化的临床药物，单独或组合。
