乌头属是多种结构多样的代谢物的主要来源，具有显着的药理学意义。本研究调查了异叶乌头 (AHS) 茎部分的代谢谱、药理学研究、抗癌潜力和分子对接分析。使用两种模式 (ESI /ESI-) 的 LC-MS/MS-orbitrap 和 EI 模式的 GC-MS 对 AHS 提取物的代谢物分析进行实验检查。体外MTT模型用于研究抗癌潜力，而体内动物模型用于研究抗炎和镇痛活性。 MOE软件用于分子对接研究。在 MeOH 提取物中鉴定出 44 种代谢物（ESI 阳性模式下 26 种，ESI 阴性模式下 18 种），共鉴定出 118 种新的和先前已知的代谢物，而在 ESI 提取物中鉴定出 74 种代谢物（ESI 模式下 46 种，ESI 阴性模式下 28 种）。通过 LCMS/MS 进行正己烷萃取。鉴定的代谢物包括24种酚类化合物、18种生物碱、10种黄酮类化合物、24种萜类化合物、2种香豆素、2种木脂素和38种其他脂肪酸和有机化合物。鉴定出的主要生物活性代谢物是大麦芽碱、赫那金、芒柄花素、白杨素、N-甲基赫那金、几内酯、姜烯酚、考拉素、colneleate、zerumbone、medicarpin、boldine、miraxinthin-v 和 lariciresinol-4-O-glucoside。此外，GC-MS 研究有助于根据质谱和保留指数鉴定挥发性和非挥发性化学成分。甲醇提取物显着抑制 H9c2 和 MDCK 癌细胞的肿瘤进展，IC50 值为 186.39 和 199.63 μg/mL。相比之下，阳性对照乌头碱对 H9c2 和 MDCK 细胞系表现出有效的 IC50 值（132.32 和 141.58 μg/mL）。抗炎（角叉菜胶引起的后爪水肿）和抗伤害（乙酸引起的扭体）作用分别显着依赖于剂量（p < 0.001）和（p < 0.05）。此外，还对抗炎酶（COX-2）（PDB ID：5JVZ）和癌酶 ADAM10（PDB ID：6BDZ）的已识别配体进行了分子对接研究，证实了其抗炎和抗癌作用。计算机模型。在所有配体中，L2、L3 和 L7 表现出最有效的抑制 COX-2 炎症的潜力，结合能为 -7.3424、-7.0427 和 -8.3562 kcal/mol。相反，针对 ADAM10 癌症蛋白，配体 L1、L4、L6 和 L7 的结合能分别为 -8.0650、-7.7276、-7.0454 和 -7.2080 kcal/mol，显示出显着的有效性。总体而言，这项 AHS 研究中揭示的已鉴定代谢物有望在化学分类学和药理学学科中发现新的可能性。© 2024 作者。由美国化学会出版。

The Aconitum genus is a leading source of a wide range of structurally diverse metabolites with significant pharmacological implications. The present study investigated metabolite profiling, pharmacological investigation, anticancer potential, and molecular docking analysis of the stem part of Aconitum heterophyllum (AHS). The metabolite profiling of the AHS extract was experimentally examined using LC-MS/MS-orbitrap in both modes (ESI+/ESI-) and GC-MS in EI mode. The in vitro MTT model was used to study the anticancer potential, while the in vivo animal model was used to study the anti-inflammatory and antinociceptive activities. The MOE software was used for the molecular docking study. A total of 118 novel and previously known metabolites, among 44 metabolites (26 in ESI+ positive mode and 18 in ESI- negative mode) in the MeOH extract, while 74 metabolites (46 in ESI+ and 28 in ESI- mode) were identified in the n-hexane extract via LCMS/MS. The identified metabolites include 24 phenolic compounds, 18 alkaloids, 10 flavonoids, 24 terpenoids, 2 coumarins, 2 lignans, and 38 other fatty acids and organic compounds. The major bioactive metabolites identified were hordenine, hernagine, formononetin, chrysin, N-methylhernagine, guineesine, shogaol, kauralexin, colneleate, zerumbone, medicarpin, boldine, miraxinthin-v, and lariciresinol-4-O-glucoside. Furthermore, the GC-MS study helped in the identification of volatile and nonvolatile chemical constituents based on the mass spectrum and retention indices. The methanol extract significantly inhibited tumor progression in H9c2 and MDCK cancer cells with IC50 values of 186.39 and 199.63 μg/mL. In comparison, the positive control aconitine exhibited potent IC50 values (132.32 and 141.58 μg/mL) against H9c2 and MDCK cell lines. The anti-inflammatory (carrageenan-induced hind paw edema) and antinociceptive (acetic acid-induced writhing) effects were significantly dose-dependent, (p < 0.001) and (p < 0.05), respectively. In addition, a molecular docking study was conducted on identified ligands against the anti-inflammatory enzyme (COX-2) (PDB ID: 5JVZ) and the cancer enzyme ADAM10 (PDB ID: 6BDZ) which confirmed the anti-inflammatory and anticancer effects in an in silico model. Among all ligands, L2, L3, and L7 exhibit the most potent potential for inhibiting COX-2 inflammation with binding energies of -7.3424, -7.0427, and -8.3562 kcal/mol. Conversely, against ADAM10 cancer protein, ligands L1, L4, L6, and L7, with binding energies of -8.0650, -7.7276, -7.0454, and -7.2080 kcal/mol, demonstrated notable effectiveness. Overall, the identified metabolites revealed in this AHS research study hold promise for discovering novel possibilities in the disciplines of chemotaxonomy and pharmacology.© 2024 The Authors. Published by American Chemical Society.