薛俊娟，葉國華，魏國棟, 2, 3，王雷清，楊 丹，李 波, 2, 3

芫花枝葉三萜類化學成分及其α-葡萄糖苷酶抑制活性研究

薛俊娟1，葉國華1，魏國棟1, 2, 3，王雷清1，楊 丹1，李 波1, 2, 3

1. 山東中醫(yī)藥高等專科學校 中藥系，山東 煙臺 264199 2. 山東省高等學校 中藥生產(chǎn)與質(zhì)量控制新技術(shù)研發(fā)中心，山東 煙臺 264199 3. 黃河流域道地藥材產(chǎn)業(yè)高質(zhì)量發(fā)展協(xié)同創(chuàng)新中心，山東 煙臺 264199

研究芫花枝葉中三萜類成分及其α-葡萄糖苷酶抑制活性。運用硅膠柱色譜、Sephadex LH-20凝膠柱色譜、半制備高效液相色譜及重結(jié)晶等多種分離方法進行分離純化，根據(jù)理化性質(zhì)以及波譜數(shù)據(jù)對分離的單體化合物進行結(jié)構(gòu)鑒定，對獲得的化合物測定α-葡萄糖苷酶抑制活性。從芫花95%乙醇提取物中共分離得到20個三萜類化合物，分別鑒定為山楂酸（1）、常春藤皂苷元（2）、齊墩果酸（3）、3β,13β-dihydroxyolean-11-en-28-oic acid（4）、11-oxoerythrodiol（5）、3β-hydroxy-11-oxo-olean-12-en-28-oic acid（6）、坡模酸（7）、3β-hydroxyolean-12-en-28-aldehyde（8）、3β-hydroxyolean-12-en-11-one（9）、3-羰基齊墩果酸（10）、古柯二醇（11）、oleanolic acid 2-oxopropyl ester（12）、熊果酸（13）、ilelatifol A（14）、3β-hydroxy-11-oxours-12-en-28-oic acid（15）、2α,3β-dihydroxyurs-12-en-28-oic acid（16）、3β-hydroxy-11-oxo-ursan-12-ene（17）、2-oxopropyl(3β)-3-hydroxyurs-12-en-28-oate（18）、熊果醛（19）和11α-methoxyurs-12-ene-3β,12-diol（20）。其中化合物1、3、4、13、15和16對α-葡萄糖苷酶具有較明顯的抑制活性。化合物6為首次從芫花中分離得到?；衔?～5、7～20為瑞香屬內(nèi)首次分離得到。化合物1、3、4、13、15和16表現(xiàn)出高于陽性對照阿卡波糖的α-葡萄糖苷酶的抑制活性，這表明芫花的枝葉中含具有降糖作用的天然活性分子。

芫花；齊墩果烷型三萜；烏蘇烷型三萜；α-葡萄糖苷酶抑制活性；山楂酸；常春藤皂苷元；齊墩果酸

芫花Sieb. et Zucc是瑞香科瑞香屬植物，多分枝，花期3～5月，果期6～7月。生于海拔300～1 000 m，宜溫暖的氣候，性耐旱怕澇，廣泛分布于河北、山西、陜西、甘肅、山東、江蘇、安徽、浙江、江西、福建、臺灣、河南、湖北、湖南、四川、貴州等省。其性溫，味辛苦，有大毒?！败净ā敝麃碓从凇渡褶r(nóng)本草經(jīng)》，在其春季花尚未開時，采其花蕾干燥后入藥，具有瀉水逐飲、解毒殺蟲等功效，常用于水腫脹滿、痰飲急劇、氣逆喘咳、二便不利等癥。芫花的根皮也可藥用，有消腫解毒、活血化瘀之效，可用于急性乳腺炎、癰癤腫毒、淋巴結(jié)結(jié)核、腹水、風濕痛、牙痛、跌打損傷等癥。芫花具有廣泛的生物活性，近年來關(guān)于其提取物如羥基芫花素、瑞香烷二萜等的研究屢見報道，芫花在抗病毒、抗腫瘤以及神經(jīng)保護等方面有明顯作用[1-4]。

目前對于芫花中活性成分的研究主要集中在花蕾和根部，為了進一步探明芫花枝葉的化學成分，開發(fā)枝葉的藥用價值，本課題著重研究芫花枝葉部位三萜類化學成分及其α-葡萄糖苷酶抑制活性，共分離鑒定20個三萜類化合物，主要包括烏蘇烷型三萜與齊墩果烷型三萜，包括山楂酸（maslinic acid，1）、常春藤皂苷元（hederagenin，2）、齊墩果酸（oleanolic acid，3）、3β,13β-dihydroxyolean-11-en-28-oic acid（4）、11-oxoerythrodiol（5）、3β-hydroxy-11-oxo-olean-12-en-28-oic acid（6）、坡模酸（pomolic acid，7）、3β-hydroxyolean-12-en-28-aldehyde（8）、3β-hydroxyolean-12-en-11-one（9）、3-羰基齊墩果酸（3-oxo-oleanolic acid，10）、古柯二醇（erythrodiol，11）、oleanolic acid 2-oxopropyl ester（12）、熊果酸（ursolic acid，13）、ilelatifol A（14）、3β-hydroxy-11-oxours-12-en-28-oic acid（15）、2α,3β-dihydroxyurs-12-en-28-oic acid（16）、3β-hydroxy-11-oxo-ursan-12-ene（17）、2-oxopropyl(3β)-3-hydroxyurs-12-en-28-oate（18）、熊果醛（ursolic aldehyde，19）和11α-methoxyurs-12-ene-3β,12-diol（20）?；衔?為首次從芫花中分離得到?；衔?～5、7～20為瑞香屬首次分離得到。其中化合物1、3、4、13、15和16表現(xiàn)出高于陽性對照阿卡波糖的α-葡萄糖苷酶抑制活性。

1 儀器與材料

Bruker Avance DRX-600型核磁共振光譜儀（德國Bruker公司）；Agilent 6545 QTOF型質(zhì)譜儀、Agilent 1260-6460型三重四極桿質(zhì)譜儀（美國Agilent公司）；島津LC-20A型型高效液相色譜儀（日本Shimadzu公司）；R-100型旋轉(zhuǎn)蒸發(fā)儀（瑞士Buchi公司）；YMC-Pack ODS-A型色譜柱（250 mm×10 mm，5 μm，日本YMC株式會社）；D101-大孔吸附樹脂（上海東鴻化工有限公司）；Sephadex LH-20（瑞典GE醫(yī)療生命科學公司）；薄層色譜硅膠板GF254及300～400目硅膠（青島海洋化工有限公司）；用于柱色譜的所有溶劑均為分析級（天津富宇精細化工有限公司）；用于HPLC的溶劑為色譜級（瑞典歐森巴克化學公司）。阿卡波糖（acarbose，批號A3230505001，質(zhì)量分數(shù)≥98%），北京索萊寶科技有限公司。

芫花于2019年6月采自山東省萊陽市，由山東中醫(yī)藥高等?？茖W校中藥系魏國棟副教授鑒定為瑞香科瑞香屬植物芫花Sieb. et Zucc的枝葉。樣品標本（2019-06-DG）保藏于山東中醫(yī)藥高等專科學校中藥系。

2 提取與分離

芫花干燥枝葉（5.0 kg），粉碎成粉末后用95%乙醇室溫提取3次（每次用溶劑40 L，7 d/次）。提取液經(jīng)減壓蒸餾得到粗浸膏（500.2 g），粗浸膏加水混懸后依次用醋酸乙酯和正丁醇溶液萃取分別萃取3次，減壓濃縮后得到醋酸乙酯浸膏（255.3 g）。將醋酸乙酯浸膏進行D-101大孔樹脂柱色譜，依次用30%、50%、80%、95%的乙醇-水梯度洗脫。80%的餾分減壓濃縮得到浸膏60.5 g，經(jīng)過硅膠柱色譜粗分離（100～200目），以石油醚-醋酸乙酯（10∶1～0∶1）梯度洗脫，得到9個組分（A～I）。

E段（2.8 g）經(jīng)過RP18反相柱色譜以甲醇-水體系（60%～100%）進行梯度洗脫，得到8個餾分E-F1～E-F8。E-F1（300.0 mg）通過半制備HPLC（85%乙腈-水）純化得到化合物12（3.3 mg，R＝20 min）和18（5.2 mg，R＝21 min）。E-F2（800.0 mg）經(jīng)過Sephadex LH-20凝膠（二氯甲烷-甲醇1∶1）色譜后通過半制備HPLC（85%乙腈-水）純化得到化合物10（2.1 mg，R＝13 min）和11（15.3 mg，R＝17 min）。E-F3（500.0 mg）經(jīng)過Sephadex LH-20凝膠（二氯甲烷-甲醇1∶1）色譜后通過半制備HPLC（75%乙腈-水）純化得到化合物17（2.8 mg，R＝10 min）和9（2.9 mg，R＝13 min）。E-F6（800.0 mg）經(jīng)過Sephadex LH-20凝膠（二氯甲烷-甲醇1∶1）柱色譜后通過半制備HPLC（68%乙腈-水）純化得到化合物8（10.1 mg，R＝20 min）、19（30.1 mg，R＝28 min）和20（4.4 mg，R＝40 min）。

F段（2.0 g）以二氯甲烷-甲醇（1∶0～100∶1）梯度洗脫得到5個餾份F-S1～F-S5。F-S4（300.0 mg）經(jīng)過Sephadex LH-20凝膠（二氯甲烷-甲醇1∶1）色譜后通過半制備HPLC（55%乙腈-水）純化得到化合物2（3.0 mg，R＝20 min）。F-S5（200.0 mg）經(jīng)過Sephadex LH-20凝膠（二氯甲烷-甲醇1∶1）色譜后通過半制備HPLC（60%乙腈-水）純化得到化合物1（3.0 mg，R＝34min）和16（5.1 mg，R＝42 min）。

G段（7.0 g）進行RP18反相柱色譜，以60%～100%的甲醇-水進行梯度洗脫得到10個部分，G-F1～F10。G-F10（800.0 mg）經(jīng)過Sephadex LH-20凝膠（100%甲醇）色譜后通過半制備液相HPLC（50%乙腈-水）純化得到化合物4（30.0 mg，R＝25 min）。G-F9（500.0 mg）經(jīng)過Sephadex LH-20凝膠（100%甲醇）色譜后通過半制備液相HPLC（69 %乙腈-水）純化得到化合物6（2.0 mg，R＝18 min）、15（2.3 mg，R＝20 min）和7（2 mg，R＝23 min）。G-F7（500.0 mg）經(jīng)過Sephadex LH-20凝膠（100%甲醇）色譜后通過半制備液相HPLC（70%乙腈-水）純化得到化合物5（7.6 mg，R＝39 min）和14（4.7 mg，R＝36 min）。

H段（5.0 g）以二氯甲烷-甲醇（1∶0～100∶1）梯度洗脫進行硅膠柱色譜分離，得到3個部分H-S1～H-S3。H-S2（3.0 g）經(jīng)過反復重結(jié)晶得到化合物13（2.0 g）。H-S2（500.0 mg）經(jīng)過Sephadex LH-20凝膠色譜柱色譜（二氯甲烷-甲醇1∶1）后經(jīng)過半制備HPLC（88%乙腈-水）純化得到化合物3（5.6 mg，R＝26.5 min）。

3 結(jié)構(gòu)鑒定

化合物1：白色粉末，ESI-MS/: 473.4 [M＋H]+，推測該化合物分子式為C30H48O4，1H-NMR (600 MHz, C5D5N): 5.49 (1H, t,= 3.8 Hz, H-12), 4.12 (1H, m, H-2), 3.42 (1H, d,= 9.3 Hz, H-3), 3.32 (1H, dd,= 14.0, 4.6 Hz, H-18), 2.27 (1H, dd,= 12.4, 4.5 Hz, H-1a), 2.10 (2H, m, H2-11), 2.08 (1H, m, H-22a), 2.02 (2H, m, H-16), 1.86 (1H, m, H-22b), 1.86 (1H, m, H-9), 1.84 (1H, m, H-19a), 1.57 (1H, m, H-6a), 1.56 (1H, m, H-7a), 1.49 (1H, m, H-21a), 1.45 (1H, m, H-6b), 1.36 (1H, m, H-7b), 1.34 (2H, m, H-1b, H-19b), 1.30 (3H, s, Me-23), 1.29 (3H, s, Me-27), 1.23 (2H, q,= 3.3 Hz, H2-15), 1.21 (1H, d,= 3.5 Hz, H-21b), 1.10 (3H, s, Me-24), 1.06 (1H, m, H-5), 1.05 (3H, s, Me-30), 1.02 (3H, s, Me-25), 1.01 (3H, s, Me-26), 0.96 (3H, s, Me-29)；13C-NMR (150 MHz, C5D5N): 180.6 (C-28), 145.3 (C-13), 122.9 (C-12), 84.2 (C-3), 69.0 (C-2), 56.3 (C-5), 48.6 (C-9), 48.2 (C-1), 47.1 (C-17), 46.8 (C-19), 42.6 (C-14), 42.4 (C-18), 40.3 (C-4), 40.2 (C-8), 38.9 (C-10), 34.6 (C-21), 33.7 (C-29), 33.6 (C-7), 33.6 (C-22), 31.4 (C-20), 29.7 (C-23), 28.7 (C-15), 26.6 (C-27), 24.3 (C-16), 24.2 (C-30), 24.1 (C-11), 19.3 (C-6), 18.1 (C-24), 17.9 (C-26), 17.3 (C-25)。以上波譜數(shù)據(jù)與文獻報道一致[5]，故鑒定化合物1為山楂酸。

化合物2：白色粉末，ESI-MS/: 473.4 [M＋H]+，推測該化合物分子式為C30H48O4。1H-NMR (600 MHz, C5D5N): 5.48 (1H, t,= 3.7 Hz, H-12), 4.19 (1H, m, H-3), 4.17 (1H, d,= 10.0 Hz, H-23a), 3.70 (1H, d,= 10.4 Hz, H-23b), 3.28 (1H, dd,= 14.0, 4.6 Hz, H-18), 2.15 (1H, m, H-15a), 2.07 (1H, m, H-16a), 2.00 (1H, m, H-22a), 1.96 (1H, m, H-16b), 1.92 (1H, m, H-11a), 1.86 (1H, m, H-2a), 1.78 (1H, m, H-19a), 1.77 (1H, m, H-22b), 1.76 (1H, m, H-9), 1.66 (1H, m, H-6a), 1.60 (1H, m, H-7a), 1.54 (1H, m, H-1a), 1.52 (1H, m, H-5), 1.43 (1H, m, H-6b), 1.41 (1H, m, H-21a), 1.27 (1H, m, H-19b), 1.26 (1H, m, H-7b), 1.22 (3H, s, Me-27), 1.18 (1H, m, H-21b), 1.14 (1H, m, H-15b), 1.08 (1H, dt,= 7.6, 3.8 Hz, H-1b), 1.04 (3H, s, Me-24), 1.03 (3H, s, Me-26), 0.98 (3H, s, Me-30), 0.95 (3H, s, Me-25), 0.91 (3H, s, Me-29), 0.98 (3H, s, Me-30)；13C-NMR (150 MHz, C5D5N): 180.5 (C-28), 145.2 (C-13), 123.0 (C-12), 73.7 (C-3), 68.2 (C-23), 49.0 (C-5), 48.5 (C-9), 47.0 (C-17), 46.8 (C-19), 43.3 (C-4), 42.6 (C-14), 42.4 (C-18), 40.1 (C-8), 39.2 (C-1), 37.6 (C-10), 34.6 (C-21), 33.6 (C-22), 33.6 (C-29), 33.4 (C-7), 31.3 (C-20), 28.7 (C-15), 28.1 (C-2), 26.5 (C-27), 24.2 (C-30), 24.1 (C-11), 24.1 (C-16), 19.0 (C-6), 17.9 (C-26), 16.4 (C-25), 13.5 (C-24)。以上波譜數(shù)據(jù)與文獻報道一致[6-7]，故鑒定化合物2為常春藤皂苷元。

化合物3：白色粉末，ESI-MS/: 455.8 [M－H]–，推測該化合物分子式為C30H48O3。1H-NMR (600 MHz, CD3OD): 5.24 (1H, t,= 3.8 Hz, H-12), 3.15 (1H, dd,= 11.6, 4.6 Hz, H-3), 2.85 (1H, dd,= 4.6, 13.7 Hz, H-18), 2.01 (1H, td,= 13.4, 4.0 Hz, H-16a), 1.16 (3H, s, Me-27), 0.97 (3H, s, Me-23), 0.94 (3H, s, Me-25), 0.94 (3H, s, Me-30), 0.91 (3H, s, Me-29), 0.82 (3H, s, Me-26), 0.78 (3H, s, Me-24), 0.75 (1H, m, H-5)；13C-NMR (150 MHz, CD3OD): 182.1 (C-28), 145.3 (C-13), 123.6 (C-12), 79.7 (C-3), 56.8 (C-5), 49.8 (C-29), 47.7 (C-17), 47.3 (C-19), 42.9 (C-14), 42.8 (C-18), 40.6 (C-8), 39.8 (C-1), 39.8 (C-4), 38.2 (C-10), 34.9 (C-21), 34.0 (C-22), 33.9 (C-7), 33.6 (C-29), 31.6 (C-20), 28.8 (C-15), 28.7 (C-23), 27.9 (C-2), 26.4 (C-27), 24.5 (C-11), 24.1 (C-16), 24.0 (C-30), 19.5 (C-6), 17.7 (C-26), 16.3 (C-24), 15.9 (C-25)。以上波譜數(shù)據(jù)與文獻報道一致[8]，故鑒定化合物3為齊墩果酸。

化合物4：白色粉末，ESI-MS/: 471.4 [M－H]–，推測該化合物分子式為C30H48O4。1H-NMR (600 MHz, CD3OD): 6.04 (1H, dd,10.4, 1.7 Hz, H-12), 5.35 (1H, dd,= 10.3, 3.2 Hz, H-11), 3.06 (1H, dd,= 11.6, 4.8 Hz, H-3), 2.17 (1H, td,= 13.3, 5.8 Hz, H-16a), 2.02 (1H, dd,= 13.8, 3.4 Hz, H-18), 1.02 (3H, s, Me-27), 0.95 (3H, s, Me-26), 0.89 (3H, s, Me-23), 0.88 (3H, s, Me-29), 0.84 (3H, s, Me-30), 0.81 (3H, s, Me-25), 0.69 (3H, s, Me-24)；13C-NMR (150 MHz, CD3OD): 182.7 (C-28), 137.4 (C-11), 127.8 (C-12), 92.1 (C-13), 79.5 (C-3), 56.0 (C-18), 54.5 (C-5), 51.8 (C-9), 45.6 (C-17), 42.9 (C-14), 42.6 (C-8), 40.0 (C-4), 39.4 (C-1), 38.2 (C-19), 37.5 (C-10), 35.2 (C-22), 33.5 (C-29), 32.3 (C-21), 32.3 (C-7), 28.3 (C-23), 28.3 (C-20), 27.7 (C-15), 26.5 (C-2), 23.9 (C-16), 22.4 (C-30), 19.6 (C-26), 18.8 (C-27), 18.7 (C-6), 18.5 (C-25), 15.7 (C-24)。以上波譜數(shù)據(jù)與文獻報道一致[9]，故鑒定化合物4為3β,13β-dihydroxyolean-11-en-28-oic acid。

化合物5：白色粉末，ESI-MS/: 457.3 [M＋H]+，推測該化合物分子式為C30H48O3。1H-NMR (600 MHz, CDCl3): 5.56 (1H, s, H-12), 3.46 (1H, d,= 10.9 Hz, H-28a), 3.22 (1H, m, H-3), 3.21 (1H, m, H-28b), 2.77 (1H, dt,= 13.6, 3.7 Hz, H-1a), 2.33 (1H, s, H-9), 2.15 (1H, dd,= 13.6, 4.6 Hz, H-18), 1.75 (1H, m, H-15a), 1.75 (1H, m, H-19a), 1.65 (2H, m, H2-2), 1.38 (3H, s, Me-27), 1.35 (1H, m, H-16a), 1.33 (1H, m, H-21a), 1.18 (1H, m, H-15b), 1.15 (1H, m, H-19b), 1.12 (3H, s, Me-25), 1.10 (3H, s, Me-26), 1.00 (3H, s, Me-26), 0.98 (1H, m, H-1b), 0.91 (3H, s, Me-30), 0.89 (3H, s, Me-29), 0.80 (3H, s, Me-24), 0.69 (1H, dd,= 11.7, 1.9 Hz, H-5)；13C-NMR (150 MHz, CDCl3): 200.3 (C-11), 169.6 (C-13), 128.4 (C-12), 78.9 (C-3), 69.8 (C-28), 61.9 (C-9), 55.1 (C-5), 45.5 (C-8), 45.0 (C-19), 43.6 (C-14), 42.8 (C-18), 39.3 (C-1), 39.3 (C-4), 37.2 (C-17), 37.1 (C-10), 34.0 (C-21), 33.1 (C-29), 32.8 (C-7), 31.2 (C-20), 30.8 (C-22), 28.2 (C-24), 27.4 (C-2), 26.0 (C-15), 23.5 (C-27), 23.5 (C-30), 21.6 (C-16), 18.7 (C-26), 17.6 (C-6), 16.5 (C-25), 15.7 (C-23)。以上波譜數(shù)據(jù)與文獻報道一致[10]，故鑒定化合物5為11-oxoerythrodiol。

化合物6：白色粉末，ESI-MS/: 469.4 [M－H]–，推測該化合物分子式為C30H46O4。1H-NMR (600 MHz, CDCl3): 5.63 (1H, s, H-12), 3.22 (1H, dd,= 10.9, 5.3 Hz, H-3), 2.96 (1H, dd,= 14.0, 4.7 Hz, H-18), 1.09 (3H, s, Me-23), 0.98 (3H, s, Me-30), 0.94(3H, s, Me-29), 0.93 (3H, s, Me-26), 0.90 (3H, s, Me-25), 0.77 (3H, s, Me-24)；13C-NMR (150 MHz, CDCl3):200.5 (C-11), 182.9 (C-28), 168.5 (C-13), 128.1 (C-12), 78.8 (C-3), 61.8 (C-9), 55.0 (C-5), 46.0 (C-17), 45.0 (C-14), 44.1 (C-19), 43.4 (C-8), 41.3 (C-18), 39.1 (C-4), 39.1 (C-1), 37.3 (C-10), 33.6 (C-21), 32.9 (C-29), 32.8 (C-7), 31.6 (C-22), 30.7 (C-2), 28.1 (C-23), 27.8 (C-15), 27.2 (C-2), 23.6 (C-27), 23.4 (C-30), 22.6 (C-16), 19.3 (C-26), 17.3 (C-6), 16.3 (C-25), 15.6 (C-24)。以上波譜數(shù)據(jù)與文獻報道一致[11]，故鑒定化合物6為3β-hydroxy-11-oxo-olean-12-en-28-oic acid。

化合物7：白色粉末，ESI-MS/: 471.4 [M－H]–，推測該化合物分子式為C30H48O4。1H-NMR (600 MHz, CD3OD): 5.29 (1H, s, H-12), 3.17 (1H, m, H-3), 2.51 (1H, s, H-18), 2.50 (1H, m, H-2a), 2.07～1.91 (2H, m, H-11), 1.93～1.79 (1H, m, H-9), 1.79～1.70 (3H, m, H-15a, 21a, 22a), 1.70～1.62 (2H, m, H-2b, 6), 1.61～1.50 (5H, m, H-1a, 1b, 16, 21b), 1.47～1.30 (2H, m, H-1b, 7b), 1.34 (3H, s, Me-27), 0.99 (3H, s, Me-23), 0.95 (3H, s, Me-25), 0.94 (3H, d,= 6.6 Hz, Me-30), 0.90 (1H, m, H-5), 0.81 (3H, s, Me-24), 0.79 (3H, s, Me-26)；13C-NMR (150 MHz, CD3OD): 140.1 (C-13), 129.4 (C-12), 79.8 (C-3), 73.6 (C-19), 56.8 (C-5), 55.1 (C-18), 43.1 (C-20), 42.6 (C-14), 41.1 (C-8), 39.9 (C-4), 39.8 (C-10), 39.1 (C-22), 34.2 (C-7), 29.6 (C-15), 28.7 (C-23), 27.9 (C-21), 27.3 (C-16), 27.1 (C-29), 26.7 (C-2), 24.8 (C-27), 24.7 (C-11), 19.6 (C-1), 17.5 (C-24), 16.6 (C-26), 16.3 (C-30), 15.9 (C-25)。以上波譜數(shù)據(jù)與文獻報道一致[12]，故鑒定化合物7為pomolic acid。

化合物8：白色粉末，ESI-MS/: 463.4 [M＋Na]+，推測該化合物分子式為C30H48O2。1H-NMR (600 MHz, CDCl3): 9.39 (1H, s, H-28), 5.34 (1H, t,= 3.7 Hz, H-12), 3.20 (1H, dd,= 11.4, 4.4 Hz, H-3), 2.62 (1H, m, H-18), 1.97 (1H, m, H-11a), 1.88 (1H, m, H-11b), 1.13 (3H, s, Me-27), 0.98 (3H, s, Me-23), 0.91 (3H, s, Me-29), 0.91 (3H, s, Me-24), 0.90 (3H, s, Me-30), 0.77 (3H, s, Me-25), 0.73 (3H, s, Me-26)；13C- NMR (150 MHz, CDCl3): 207.7 (C-28), 143.1 (C-13), 123.4 (C-12), 79.1 (C-3), 55.3 (C-5), 49.2 (C-9), 47.7 (C-17), 45.7 (C-19), 41.8 (C-14), 40.5 (C-18), 39.7 (C-8), 38.9 (C-4), 38.6 (C-1), 37.1 (C-10), 33.3 (C-21), 33.2 (C-7), 32.9 (C-22), 30.8 (C-29), 28.2 (C-20), 27.9 (C-2), 27.3 (C-23), 26.9 (C-15), 25.7 (C-27), 23.6 (C-16), 23.6 (C-30), 22.2 (C-11), 18.4 (C-6), 17.2 (C-26), 15.7 (C-24), 15.5 (C-15)。以上波譜數(shù)據(jù)與文獻報道一致[13]，故鑒定化合物8為3β-hydroxyolean-12-en-28-aldehyde。

化合物9：白色粉末，ESI-MS/: 441.6 [M＋H]+，推測該化合物分子式為C30H48O2。1H-NMR (600 MHz, CDCl3): 5.58 (1H, s, H-12), 3.23 (1H, dd,= 11.1, 5.3 Hz, H-3), 2.79 (1H, dt,= 13.5, 3.6 Hz, H-9), 1.36 (3H, s, Me-27), 1.13 (3H, s, Me-24), 1.13 (3H, s, Me-23), 1.00 (3H, s, Me-26), 0.90 (3H, s, Me-29), 0.88 (3H, s, Me-30), 0.86 (3H, s, Me-25), 0.80 (3H, s, Me-28)；13C-NMR (150 MHz, CDCl3): 200.5 (C-11), 170.8 (C-13), 128.3 (C-12), 78.9 (C-3), 61.9 (C-9), 55.1 (C-5), 47.7 (C-18), 45.6 (C-14), 45.3 (C-19), 43.5 (C-8), 39.3 (C-1), 37.2 (C-4), 36.7 (C-22), 34.6 (C-21), 33.2 (C-29), 32.9 (C-7), 32.5 (C-17), 31.2 (C-6), 28.9 (C-28) 28.2 (C-23), 27.5 (C-2), 26.6 (C-16), 26.5 (C-15), 23.6 (C-30), 23.6 (C-27), 18.9 (C-6), 17.6 (C-26), 16.5 (C-24), 15.7 (C-25)。以上波譜數(shù)據(jù)與文獻報道一致[14]，故鑒定化合物9為3β-hydroxyolean-12-en-11-one。

化合物10：白色粉末，ESI-MS/: 477.8 [M＋Na]+，推測該化合物分子式為C30H46O3。1H-NMR (600 MHz, CDCl3): 5.29 (1H, t,= 3.7 Hz, H-12), 2.83 (1H, dd,= 13.8, 4.7 Hz, H-18), 2.54 (1H, ddd,= 15.8, 11.1, 7.3 Hz, H-2a), 2.36 (1H, ddd,= 15.9, 6.8, 3.6 Hz, H-2b), 1.14 (3H, s, Me-27), 1.08 (3H, s, Me-23), 1.04 (3H, s, Me-24), 1.02 (3H, s, Me-25), 0.92 (3H, s, Me-29), 0.90 (3H, s, Me-30), 0.80 (3H, s, Me-26)；13C-NMR (150 MHz, CDCl3): 217.9 (C-3), 184.4 (C-28), 143.8 (C-13), 122.5 (C-12), 55.4 (C-5), 47.6 (C-4), 47.0 (C-9), 46.7 (C-17), 45.9 (C-19), 41.8 (C-14), 41.1 (C-18), 39.4 (C-1), 39.2 (C-8), 36.9 (C-10), 34.3 (C-2), 33.9 (C-21), 33.2 (C-29), 32.5 (C-22), 32.3 (C-7), 30.8 (C-20), 27.8 (C-15), 26.6 (C-24), 26.0 (C-27), 23.7 (C-30), 23.6 (C-11), 23.0 (C-16), 21.6 (C-23), 19.7 (C-6), 17.1 (C-26), 15.1 (C-25)。以上波譜數(shù)據(jù)與文獻報道一致[15]，故鑒定化合物10為3-羰基齊墩果酸。

化合物11：白色粉末，ESI-MS/: 465.4 [M＋Na]+，推測該化合物分子式為C30H50O2。1H-NMR (600 MHz, CDCl3): 5.19 (1H, t,= 3.7 Hz, H-12), 3.55 (1H, d,= 11.0 Hz, H-28a), 3.22 (1H, m, H-3), 3.21 (1H, d,= 11.3 Hz, H-28b), 1.98 (1H, dd,= 13.5, 4.7 Hz, H-18), 1.92～1.83 (3H, m, H2-11, 16α), 1.75～1.68 (2H, m, H-19α, 15b), 1.17 (3H, s, Me-27), 1.00 (3H, s, Me-23), 0.94 (3H, s, Me-26), 0.93 (3H, s, Me-25), 0.89 (3H, s, Me-29), 0.87 (3H, s, Me-30), 0.79 (3H, s, Me-24)；13C-NMR (150 MHz, CDCl3):144.3 (C-13), 122.5 (C-12), 79.2 (C-3), 69.9 (C-28), 55.3 (C-5), 47.7 (C-9), 46.6 (C-19), 42.5 (C-18), 41.9 (C-14), 39.9 (C-8), 38.9 (C-4), 38.7 (C-1), 37.1 (C-10), 37.1 (C-17), 34.2 (C-21), 33.3 (C-29), 32.7 (C-7), 31.2 (C-22), 31.1 (C-20), 28.2 (C-23), 27.4 (C-2), 26.1 (C-27), 25.7 (C-15), 23.73 (C-30), 23.67 (C-11), 22.1 (C-16), 18.5 (C-6), 16.9 (C-26), 15.7 (C-24), 15.7 (C-25)。以上波譜數(shù)據(jù)與文獻報道一致[16]，故鑒定化合物11為古柯二醇。

化合物12：白色粉末，ESI-MS/: 535.4 [M＋Na]+，推測該化合物分子式為C33H52O4。1H-NMR (600 MHz, CDCl3): 5.29 (1H, t,= 3.7 Hz, H-12), 4.59 (1H, d,= 16.8 Hz, H-1￠a), 4.54 (1H, d,= 16.7, H-1￠b), 3.21 (1H, dd,= 11.3, 4.3 Hz, H-3), 2.87 (1H, dd,= 13.8, 4.7 Hz, H-18), 2.16 (3H, s, 3￠-Me), 1.14 (3H, s, 27-Me), 0.99 (3H, s, 23-Me), 0.93 (3H, s, 29-Me), 0.91 (3H, s, Me-24), 0.90 (3H, s, Me-30), 0.78 (3H, s, Me-25), 0.72 (3H, s, Me-26)；13C-NMR (150 MHz, CDCl3): 202.5 (C-2￠), 177.2 (C-28), 143.7 (C-13), 122.7 (C-12), 79.2 (C-3), 68.2 (C-1￠), 55.4 (C-5), 47.8 (C-9), 47.0 (C-17), 46.1 (C-19), 41.9 (C-14), 41.4 (C-18), 39.5 (C-8), 38.9 (C-4), 38.6 (C-1), 37.2 (C-10), 34.0 (C-21), 33.2 (C-30), 32.8 (C-22), 32.5 (C-7), 30.8 (C-20), 28.2 (C-23), 27.8 (C-15), 27.3 (C-2), 26.5 (C-3￠), 26.0 (C-27), 23.8 (C-29), 23.6 (C-16), 23.3 (C-11), 18.5 (C-6), 17.1 (C-26), 15.7 (C-25), 15.5 (C-24)。以上波譜數(shù)據(jù)與文獻報道一致[17]，故鑒定化合物12為oleanolic acid 2-oxopropyl ester。

化合物13：白色粉末，ESI-MS/: 457.4 [M＋H]+，推測該化合物分子式為C30H48O3。1H NMR (600 MHz, CD3OD): 5.23 (1H, t,= 3.7 Hz, H-12), 3.15 (1H, dd,= 11.7, 4.6 Hz, H-3), 2.20 (1H, dd,= 11.4, 1.8 Hz, H-18), 1.12 (3H, s, Me-23), 0.98 (3H, s, Me-27), 0.96 (3H, d,= 5.0 Hz, Me-29), 0.96 (3H, s, Me-26), 0.89 (3H, d,= 6.5 Hz, Me-30), 0.85 (3H, s, Me-24), 0.78 (3H, s, Me-25)；13C-NMR (150 MHz, CD3OD): 181.7 (C-28), 139.6 (C-13), 126.9 (C-12), 79.7 (C-3), 56.7 (C-5), 54.4 (C-18), 49.0 (C-9), 43.2 (C-14), 40.8 (C-8), 40.4 (C-19), 40.0 (C-1), 39.8 (C-4), 38.1 (C-10), 38.1 (C-22), 34.3 (C-7), 31.8 (C-21), 29.2 (C-15), 28.8 (C-23), 27.9 (C-2), 25.3 (C-16), 24.4 (C-11), 24.1 (C-27), 21.6 (C-30), 19.5 (C-6), 17.8 (C-29), 17.7 (C-26), 16.4 (C-24), 16.0 (C-25)。以上波譜數(shù)據(jù)與文獻報道一致[18-19]，故鑒定化合物13為熊果酸。

化合物14：白色粉末，ESI-MS/: 457.3 [M＋H]+，推測該化合物分子式為C30H48O3。1H-NMR (600 MHz, CDCl3):5.51 (1H, s, H-12), 3.46 (1H, d,= 10.9 Hz, H-28a), 3.22 (1H, dd,= 11.4, 4.9 Hz, H-3), 3.16 (1H, d,= 11.0 Hz, H-28b), 2.32 (1H, s, H-9), 1.32 (3H, s, Me-27), 1.16 (3H, s, Me-23), 1.15 (3H, d,= 6.9 Hz, Me-30), 0.96 (3H, s, Me-24), 0.82 (3H, d,= 6.5 Hz, Me-29), 0.81 (3H, s, Me-25)；13C-NMR (150 MHz, CDCl3): 199.8 (C-11), 163.8 (C-13), 130.7 (C-12), 78.9 (C-3), 69.9 (C-28), 61.7 (C-9), 55.0 (C-5), 54.1 (C-18), 45.2 (C-8), 43.8 (C-14), 39.4 (C-20), 39.4 (C-1), 39.3 (C-4), 39.1 (C-19), 38.5 (C-17), 37.1 (C-10), 35.0 (C-22), 32.9 (C-7), 30.4 (C-21), 28.2 (C-23), 27.4 (C-2), 26.7 (C-15), 22.8 (C-16), 21.3 (C-30), 20.7 (C-27), 18.5 (C-26), 17.7 (C-6), 17.5 (C-29), 16.7 (C-25), 15.7 (C-24)。以上波譜數(shù)據(jù)與文獻報道一致[20-21]，故鑒定化合物14為ilelatifol A。

化合物15：白色粉末，ESI-MS/: 505.4 [M＋Cl]–，推測該化合物分子式為C30H46O4。1H-NMR (600 MHz, CDCl3): 5.56 (1H, s, H-12), 1.31 (3H, s, Me-27), 1.12 (3H, s, Me-25), 0.99 (3H, s, Me-24), 0.97 (3H, s, Me-20), 0.97 (3H, d,= 6.2, Hz, Me-30), 0.87 (3H, d,= 6.4 Hz, Me-29), 0.79 (3H, s, Me-5)；13C-NMR (150 MHz, CDCl3): 200.5 (C-11), 179.7 (C-28), 163.7 (C-13), 130.6 (C-12), 78.7 (C-3), 61.5 (C-9), 55.0 (C-5), 52.9 (C-18), 47.4 (C-17), 44.7 (C-8), 43.9 (C-14), 39.2 (C-1), 39.1 (C-4), 38.7 (C-20), 38.6 (C-19), 37.2 (C-10), 36.1 (C-22), 33.1 (C-7), 30.4 (C-21), 28.5 (C-15), 28.1 (C-23), 27.1 (C-2), 23.9 (C-16), 21.0 (C-27), 21.0 (C-30), 18.9 (C-26), 17.5 (C-6), 17.1 (C-29), 16.3 (C-25), 15.6 (C-24)。以上波譜數(shù)據(jù)與文獻報道一致[22-23]，故鑒定化合物15為3β-hydroxy-11-oxours-12-en-28-oic acid。

化合物16：白色粉末，ESI-MS/: 473.8 [M＋H]+，推測該化合物分子式為C30H48O4。1H-NMR (600 MHz, C5D5N): 5.46 (1H, t, H-12), 4.08 (1H, m, H-2), 3.39 (1H, d,= 9.5 Hz, H-3), 2.62 (1H, d,= 11.5 Hz, H-18), 1.25 (3H, s, Me-23), 1.18 (3H, s, Me-27), 1.06 (3H, s, Me-26), 1.05 (3H, s, Me-24), 0.98 (3H, d,= 6.5 Hz, Me-30), 0.97 (3H, s, Me-25), 0.93 (3H, d,= 6.5 Hz, Me-29)；13C-NMR (150 MHz, C5D5N): 180.3 (C-28), 139.7 (C-13), 125.9 (C-12), 84.2 (C-3), 68.9 (C-2), 56.3 (C-5), 53.9 (C-18), 48.4 (C-1), 48.4 (C-17), 48.4 (C-9), 42.9 (C-14), 40.4 (C-4), 40.2 (C-8), 39.8 (C-19), 39.8 (C-20), 38.8 (C-10), 37.8 (C-22), 33.9 (C-7), 31.4 (C-16), 29.7 (C-21), 29.0 (C-23), 25.3 (C-15), 24.3 (C-27), 24.1 (C-11), 21.8 (C-29), 19.2 (C-6), 18.1 (C-24), 17.9 (C-30), 17.8 (C-26), 17.3 (C-25)。以上波譜數(shù)據(jù)與文獻報道一致[24]，故鑒定化合物16為2α,3β-dihydroxyurs-12-en-28-oic acid。

化合物17：白色粉末，ESI-MS/: 441.4 [M＋H]+，推測該化合物分子式為C30H48O2。1H-NMR (600 MHz, CDCl3): 5.54 (1H, s, H-12), 3.23 (1H, dd,= 11.4, 5.0 Hz, H-3), 2.32 (1H, s, H-9), 1.67 (1H, m, H-18), 1.29 (3H, s, Me-27), 1.17 (3H, s, Me-26), 1.00 (3H, s, Me-25), 0.94 (3H, s, Me-24), 0.81 (3H, s, Me-23), 0.81 (3H, d,= 3.9 Hz, Me-30), 0.80 (3H, d,= 6.5 Hz, Me-29)；13C-NMR (150 MHz, CDCl3): 200.0 (C-1), 165.1 (C-13), 130.6 (C-12), 79.0 (C-3), 61.7 (C-9), 59.1 (C-18), 55.1 (C-5), 45.3 (C-14), 43.8 (C-8), 41.1 (C-22), 39.4 (C-20), 39.4 (C-1), 39.4 (C-19), 39.3 (C-4), 37.1 (C-10), 34.1 (C-17), 33.0 (C-7), 31.0 (C-21), 29.0 (C-28), 28.2 (C-23), 27.7 (C-16), 27.5 (C-2), 27.4 (C-15), 21.3 (C-30), 20.7 (C-27), 18.7 (C-26), 17.7 (C-6), 17.6 (C-29), 16.7 (C-25), 15.7 (C-24)。以上波譜數(shù)據(jù)與文獻報道一致[25]，故鑒定化合物17為3β-hydroxy-11-oxo-ursan-12-ene。

化合物18：白色粉末，ESI-MS/: 513.8 [M＋H]+，推測該化合物分子式為C33H52O4。1H-NMR (600 MHz, CDCl3):5.26 (1H, d,= 3.1 Hz, H-12), 4.54 (2H, m, H2-1￠), 3.22 (1H, dd,= 11.4, 4.7 Hz, H-3), 2.15 (3H, s, Me-3￠), 1.09 (3H, s, Me-27), 0.99 (3H, s, Me-23), 0.95 (3H, d,= 6.4 Hz, Me-30), 0.92 (3H, s, Me-25), 0.86 (3H, d,= 6.7 Hz, Me-29), 0.78 (3H,s, Me-24), 0.74 (3H, s, Me-26)；13C-NMR (150 MHz, CDCl3): 202.6 (C-2￠), 177.0 (C-28), 138.1 (C-13), 125.9 (C-12), 79.2 (C-3), 68.2 (C-1￠), 55.4 (C-5), 53.0 (C-18), 48.4 (C-17), 47.7 (C-9), 42.2 (C-14), 39.7 (C-8), 39.2 (C-19), 39.0 (C-20), 38.9 (C-1), 38.8 (C-4), 37.1 (C-10), 36.8 (C-22), 33.1 (C-7), 30.8 (C-21), 28.3 (C-23), 28.1 (C-15), 27.4 (C-2), 26.5 (C-3￠), 24.4 (C-16), 23.7 (C-27), 23.4 (C-11), 21.3 (C-30), 18.4 (C-6), 17.2 (C-29), 15.8 (C-24), 15.6 (C-25)。以上波譜數(shù)據(jù)與文獻報道一致[26]，故鑒定化合物18為2-oxopropyl(3β)-3-hydroxyurs-12-en-28-oate。

化合物19：白色粉末，ESI-MS/: 441.7 [M＋H]+，推測該化合物分子式為C30H48O2。1H-NMR (600 MHz, CDCl3): 9.32 (1H, s, H-28), 5.31 (1H, t,= 3.7 Hz, H-12), 3.21 (1H, dd,= 11.3, 4.7 Hz, H-3), 1.09 (3H, s, Me-27), 0.99 (3H, s, Me-23), 0.96 (3H, d,= 6.3 Hz, Me-30), 0.92 (3H, s, Me-24), 0.88 (3H, d,= 6.5 Hz, Me-29), 0.78 (3H, s, Me-25), 0.77 (3H, s, Me-26)；13C-NMR (150 MHz, CDCl3): 207.6 (C-28), 137.9 (C-13), 126.3 (C-12), 79.1 (C-3), 55.3 (C-5), 52.7 (C-18), 50.3 (C-17), 47.7 (C-9), 42.3 (C-14), 39.9 (C-8), 39.1 (C-19), 38.9 (C-20), 38.9 (C-4), 38.8 (C-1), 37.0 (C-10), 33.2 (C-22), 32.0 (C-7), 30.3 (C-21), 28.3 (C-15), 27.3 (C-2), 27.0 (C-23), 23.4 (C-16), 23.3 (C-11), 23.3 (C-27), 21.2 (C-30), 18.4 (C-6), 17.3 (C-26), 16.8 (C-29), 15.8 (C-24), 15.6 (C-25)。以上波譜數(shù)據(jù)與文獻報道一致[27]，故鑒定化合物19為熊果醛。

化合物20：白色粉末，ESI-MS/: 507.2 [M＋Cl]–，推測該化合物分子式為C31H52O3。1H-NMR (600 MHz, CDCl3):4.53 (1H, s, 12-OH), 4.25 (1H, d,= 10.3 Hz, H-11), 3.24 (1H, q,= 8.1, 5.6 Hz, H-3), 3.17 (3H, s, 11-OMe), 2.23 (1H, m, H-18), 1.87 (1H, d,= 10.4 Hz, H-9), 1.79 (1H, td,= 13.5, 5.0 Hz, H-15a), 1.20 (3H, s, Me-27), 1.11 (3H, s, Me-25), 1.11 (3H, s, Me-26), 1.02 (3H, s, Me-23), 0.93 (3H, d,= 6.4 Hz, Me-30), 0.92 (3H, d,= 6.4 Hz, Me-29), 0.82 (3H, s, Me-24), 0.81 (3H, s, Me-28)；13C-NMR (150 MHz, CDCl3): 142.2 (C-12), 118.4 (C-13), 78.8 (C-3), 76.8 (C-11), 55.6 (C-5), 51.5 (11-OMe), 47.8 (C-18), 46.4 (C-9), 43.0 (C-14), 41.8 (C-22), 40.9 (C-19), 40.7 (C-8), 39.6 (C-20), 39.3 (C-4), 39.1 (C-1), 38.5 (C-10), 34.4 (C-7), 33.5 (C-17), 31.4 (C-21), 28.7 (C-28), 28.5 (C-23), 27.7 (C-16), 27.6 (C-2), 27.3 (C-15), 24.0 (C-27), 21.4 (C-30), 18.4 (C-6), 18.2 (C-26), 17.1 (C-29), 16.3 (C-25), 15.9 (C-24)。以上波譜數(shù)據(jù)與文獻報道一致[28]，故鑒定化合物20為11α-methoxyurs-12-ene-3β,12-diol。

4 活性測試

采用Omar等[29]的方法測試化合物1～20對α-葡萄糖苷酶的抑制活性。測試化合物在100mmol/L時對α-葡萄糖苷酶活性的抑制率，對抑制率>50%的化合物進行半數(shù)抑制濃度（median inhibition concentration，IC50）值測定。其中化合物1、3、4、13、15和16對α-葡萄糖苷酶的抑制率＞50%?；衔?、3、4、13、15和16測試IC50值：用倍比稀釋法設置濃度范圍為100、50、25、12.5、6.25、3.13、1.56 μmol/L，通過SPSS 18.0軟件中的probit回歸法計算得到IC50值分別為（15.75±0.40）、（34.18±2.26）、（20.55±1.35）、（24.76±0.21）、（34.74±0.54）、（35.60±1.66）μmol/L，抑制活性均高于陽性對照阿卡波糖[IC50為（536.9±24.0）μmol/L]。其中，化合物1、3、4為齊墩果烷型三萜，化合物13、15、16為烏蘇烷型三萜，化合物1的IC50值最小，對α-葡萄糖苷酶的抑制活性最為顯著?？梢钥闯?，在同個骨架中，化合物1相比其他2個化合物，其C-2上多連了1個α-羥基，而化合物4的羥基在C-13上，說明2位上的羥基也許由于化學位阻相較于13位羥基的更小，所以能夠與α-葡萄糖苷酶的結(jié)合更為緊密，從而更好地抑制起發(fā)揮作用。同時，20位的偕二甲基對于抑制活性也有發(fā)揮一定作用的可能性。

5 討論

本課題對芫花枝葉部分的三萜類化學成分及α-葡萄糖苷酶抑制活性進行了研究，從芫花干燥枝葉的95%乙醇提取物的醋酸乙酯部位中分離鑒定了20個三萜類化合物。芫花的天然產(chǎn)物中，有從花蕾中分離得到的具有抗腫瘤活性顯著的瑞香烷型二萜，也有從根部分離得到的具有神經(jīng)保護活性的愈創(chuàng)木烷型倍半萜，而鮮少對其枝葉進行系統(tǒng)的化學成分研究。本課題則對芫花的枝葉部分進行了系統(tǒng)的三萜類成分的分離鑒定，并得到了具有抑制α-葡萄糖苷酶的活性化合物。但本研究仍不夠深入，后續(xù)應進一步分離得到更多此結(jié)構(gòu)類型的成分以及活性化合物，即可進一步探討其構(gòu)效關(guān)系；同時也可能挖掘到結(jié)構(gòu)新穎的化合物從而擴大天然產(chǎn)物庫，后期將對其他極性部位和藥理作用展開進一步研究，以便為芫花的綜合利用提供科研數(shù)據(jù)和理論基礎(chǔ)。

利益沖突 所有作者均聲明不存在利益沖突

[1] Gupta D P, Park S H, Lee Y S,.flower extract promotes the neuroprotective effects of microglia [J]., 2023, 108: 154486.

[2] Wu J, Qiu Z Z, Wei P H,. Study on the mechanism of potential pharmacological action ofof anti-tumor based on data mining, network pharmacology and molecular docking [J].:, 2021, 2004(1): 012010.

[3] 宗明月, 張慶然, 王璐瓊, 等. 基于網(wǎng)絡藥理學和分子對接法研究芫花抗炎作用機制 [J]. 煙臺大學學報: 自然科學與工程版, 2021, 34(2): 178-185.

[4] 宗明月. 芫花根中倍半萜類成分的抗炎活性及機制研究 [D]. 煙臺: 煙臺大學, 2021.

[5] Lia Z J, Wan C P, Cai L,. Terpenoids with cytotoxic activity from the branches and leaves of[J]., 2015, 13: 246-251.

[6] Joshi B S, Singh K L, Roy R. Complete assignments of 1H and13C NMR spectra of the pentacyclic triterpene hederagenin fromLinn [J]., 1999, 37(4): 295-298.

[7] Mizui F M, Kasai R, Ohtani K,. Saponins from brans of quinoa,Willd. I [J]., 1988, 36(4): 1415-1418.

[8] Kishikawa A, Amen Y, Shimizu K. Anti-allergic triterpenes isolated from olive milled waste [J]., 2017, 69(2): 307-315.

[9] Misra L, Laatsch H. Triterpenoids, essential oil and photo-oxidative 28: > 13-lactonization of oleanolic acid from[J]., 2000, 54(8): 969-974.

[10] Kim D K, Nam I Y, Kim J W,. Pentacyclic triterpenoids from[J]., 2002, 25(5): 617-620.

[11] Zhang Q, Lu Z, Li X,. Triterpenoids and steroids from the leaves of[J]., 2015, 51(1): 178-180.

[12] Thao T T P, Chi N L, Luu N T,. Phytochemistry and anti-inflammatory activity of iridoids fromcollected in the mangrove forest of Phu Loc district, Thua Thien Hue Province, Vietnam [J]., 2021, 59(6): 943-950.

[13] Monaco P, Caputo R, Palumbo G,. Triterpenes from the galls of[J]., 1973, 12(10): 2533-2534.

[14] Herath H, Athukoralage P. Oleanane triterpenoids from[J]., 1998, 4: 253-256.

[15] Kwon H C, Lee K R, Zee O P. Cytotoxic constituents of[J]., 1997, 20(2): 180-183.

[16] Lee C K, Chang M H. The chemical constituents from the heartwood of[J]., 2000, 47(3): 555-560.

[17] Cao F, Gao Y H, Wang M,. Propylene glycol-linked amino acid/dipeptide diester prodrugs of oleanolic acid for PepT1-mediated transport: Synthesis, intestinal permeability, and pharmacokinetics [J]., 2013, 10(4): 1378-1387.

[18] Liu M C, Hu D Y, Song B AChemical constituents from(Oliv.) Havil [J]., 2011, 23(6): 1058-1060.

[19] Liu M C, Yang S J, Jin L H,. Chemical constituents of the ethyl acetate extract ofchinensis (L.) DC roots and their antitumor activities [J]., 2012, 17(5): 6156-6169.

[20] Nishimura K, Miyase T, Noguchi HAcyl-CoA: cholesterol acyltransferase (ACAT) inhibitors fromspp[J]., 2000, 54(6): 297-305.

[21] Ali M S, Ibrahim S A, Jalil S,. Ursolic acid: A potent inhibitor of superoxides produced in the cellular system [J]., 2007, 21(6): 558-561.

[22] Niesen A, Barthel A, Kluge R,. Antitumoractive endoperoxides from triterpenes [J]., 2009, 342(10): 569-576.

[23] Seebacher W, Simic N, Weis R,. Complete assignments of1H and13C NMR resonances of oleanolic acid, 18α-oleanolic acid, ursolic acid and their 11-oxo derivatives [J]., 2003, 41(8): 636-638.

[24] 莫小宇, 麥景標. 生藤乙酸乙酯部位的化學成分研究 [J]. 中國藥房, 2013, 24(15): 1409-1410.

[25] 黃火強, 閆美娜, 樸香蘭, 等. 三角葉鳳毛菊中的三萜化合物 [J]. 中國實驗方劑學雜志, 2011, 17(16): 50-53.

[26] Ding Q, Wang X H. Modulators of ROR-gamma receptors, composition and use thereof: US20170298090 [P]. 2017-10-19.

[27] Kim D H, Han K M, Chung I S,. Triterpenoids from the flower ofK. Schum. as human acyl-CoA: Cholesterol acyltransferase inhibitors [J]., 2005, 28(5): 550-556.

[28] Kaweetripob W, Mahidol C, Prawat H,. Lupane, friedelane, oleanane, and ursane triterpenes from the stem ofGriff [J]., 2013, 96: 404-417.

[29] Omar R, Li L, Yuan Tα-Glucosidase inhibitory hydrolyzable tannins fromseeds [J]., 2012, 75(8): 1505-1509.

Chemical constituents from branches and leaves ofand their α-glucosidase inhibitory activities

XUE Junjuan1, YE Guohua1, WEI Guodong1, 2, 3, WANG Leiqing1, YANG Dan1, LI Bo1, 2, 3

1. Department of Traditional Chinese Medicine, Shandong College of Traditional Chinese Medicine, Yantai 264199, China 2. New Technology Research and Development Center of Traditional Chinese Medicine Production and Quality Control in Universities of Shandong Province, Yantai 264199, China 3. Collaborative Innovation Center for High-quality Development of Genuine Medicinal Materials Industry in the Yellow River Basin, Yantai 264199, China

To investigate the triterpenoid constituents and their inhibitory effect on α-glucosidase found in the branches and leaves of Yuanhua (Sieb. et Zucc).The 95% ethanol extract ofwas separated and purified by silica gel, Sephadex LH-20, semi-preparative HPLC and other separation methods, and the structures of the isolates were identified according to the physical and chemical properties as well as spectral data. The inhibitory effect on α-glucosidase of the obtained compounds was examined.A total of 20 triterpenoids were elucidated as maslinic acid (1), hederagenin (2), oleanolic acid (3), 3β,13β-dihydroxyolean-11-en-28-oic acid (4), 11-oxoerythrodiol (5), 3β-hydroxy-11-oxo-olean-12-en-28-oic acid (6), pomolic acid (7), 3β-hydroxyolean-12-en-28-aldehyde (8), 3β-hydroxyolean-12-en-11-one (9), 3-oxo-oleanolic acid (10), erythrodiol (11), oleanolic acid 2-oxopropyl ester (12), ursolic acid (13), ilelatifol A (14), 3β-hydroxy-11-oxours-12-en-28-oic acid (15), 2α,3β-dihydroxyurs-12-en-28-oic acid (16), 3β-hydroxy-11-oxo-ursan-12-ene (17), 2-oxopropyl(3β)-3-hydroxyurs-12-en-28-oate (18), ursolic aldehyde (19) and 11α-methoxyurs-12-ene-3β,12-diol (20). Amongst, compounds 1, 3, 4, 13, 15 and 16 were observed with good inhibitory activity against α-glucosidase.Compound 6 was reported fromSieb. et Zucc for the first time. Compounds 1-5 and 7-20 were reported fromgenus for the first time. Compounds 1, 3, 4, 13, 15 and 16 displayed superior inhibitory activity compared to the positive control acarbose against α-glucosidase. This suggests the presence of bioactive constituents with hypoglycemic activity in the branches and leaves of.

Sieb. et Zucc; oleanane triterpenoid; ursane triterpenoid; α-glucosidase inhibitory activity; maslinic acid; hederagenin; oleanolic acid

R284.1

A

0253 - 2670(2024)08 - 2524 - 09

10.7501/j.issn.0253-2670.2024.08.003

2024-01-24

國家自然科學基金資助項目（22177016）；山東省中醫(yī)藥科技發(fā)展計劃項目（2020M037）；2019年醫(yī)療服務與保障能力提升補助資金“全國中藥資源普查項目”（財社［2019］39號）

薛俊娟（1979—），女，講師，碩士，從事天然藥物化學研究。E-mail: 304132729@qq.com

[責任編輯 王文倩]