張東東，黃仕其，劉媛媛，孫 玉，李玉澤，張化為，鄧 翀，宋小妹，王 薇

華中五味子根皮中1個(gè)新的菖蒲烯類倍半萜

張東東，黃仕其，劉媛媛，孫 玉，李玉澤，張化為，鄧 翀，宋小妹，王 薇*

陜西中醫(yī)藥大學(xué)藥學(xué)院，陜西 咸陽(yáng) 712046

研究華中五味子根皮的化學(xué)成分及其細(xì)胞毒活性。采用硅膠柱色譜、sephadex LH-20葡聚糖凝膠柱色譜和半制備液相色譜進(jìn)行分離純化，結(jié)合NMR、MS和ECD等數(shù)據(jù)進(jìn)行化合物結(jié)構(gòu)鑒定，采用MTT法測(cè)定化合物的細(xì)胞毒活性。從華中五味子根皮中分離得到11個(gè)化合物，分別鑒定為(7,10,11)-2,11,12-三羥基菖蒲烯（1）、五味子醇丁（2）、12-demethylwuweilignan I（3）、戈米辛M1（4）、異戈米辛O（5）、schisphenlignan C（6）、marlignan F（7）、wilsonilignan A（8）、sphenlignan A（9）、schinegllignan A（10）、schinegllignan B（11），化合物2～5和7對(duì)人肺癌A549細(xì)胞、人結(jié)直腸癌HCT116細(xì)胞和SW620細(xì)胞具有細(xì)胞毒活性，半數(shù)抑制濃度（median inhibition concentration，IC50）值為12.1～38.7 μmol/L?；衔?為新的倍半萜類化合物，命名為華中五味子倍半萜A；化合物2為新天然產(chǎn)物，化合物3～5、7、8、10～11首次從華中五味子中分離得到，化合物2～5和7具有細(xì)胞毒活性。

華中五味子；華中五味子倍半萜A；五味子醇?。桓昝仔罬1；異戈米辛O；木脂素；細(xì)胞毒活性

華中五味子根皮，又名長(zhǎng)勝七，是陜西秦巴地區(qū)著名“太白七藥”之一，其來(lái)源為五味子科（Schisandraceae）五味子屬M(fèi)ichx.華中五味子Rehd. et Wils的干燥根皮，主要分布于秦嶺山脈和大巴山區(qū)各地，具有健脾化濕、澀精止瀉等功效，主要用于消化不良，肺虛咳嗽等病癥[1]。華中五味子作為我國(guó)傳統(tǒng)中藥，又稱“南五味子”，其化學(xué)成分主要為木脂素、三萜、倍半萜以及黃酮等[2-4]，現(xiàn)代藥理研究表明其提取物和單體化合物具有保肝、抗炎、抗腫瘤、抗氧化、免疫抑制等多種藥理作用[3-6]，目前對(duì)于華中五味子植物化學(xué)成分的研究主要集中在果實(shí)和藤莖等地上部位，而對(duì)于根部位的研究較少，為進(jìn)一步研究華中五味子根皮的化學(xué)成分，本研究在前期實(shí)驗(yàn)基礎(chǔ)之上[3-4]繼續(xù)對(duì)其乙醇提取物的醋酸乙酯部位進(jìn)行系統(tǒng)分離，從中共鑒定了11個(gè)化合物，分別為(7,10,11)-2,11,12-三羥基菖蒲烯[(7,10,11)- 2,11,12-trihydroxycalamenene，1]、五味子醇?。╯chisandrol D，2）、12-demethyl- wuweilignan I（3）、戈米辛M1（gomisin M1，4）、異戈米辛O（isogomisin O，5）、schisphenlignan C（6）、marlignan F（7）、wilsonilignan A（8）、sphenlignan A（9）、schinegllignan A（10）、schinegllignan B（11）。其中化合物1為新的倍半萜類化合物，命名為華中五味子倍半萜A，化合物2為新的天然產(chǎn)物，命名為五味子醇丁，化合物3～5、7、8、10～11為首次從該植物中分離得到。細(xì)胞毒活性篩選結(jié)果發(fā)現(xiàn)，化合物2～5和7具有細(xì)胞毒活性。

1 儀器與試藥

IS50傅里葉變換紅外光譜儀（Thermo Nicolet公司）；Chirascan CD圓二色譜儀（英國(guó)應(yīng)用光物理公司）；SGW-3自動(dòng)旋光儀（上海精密儀器儀表有限公司）；Burker AVANCE 400核磁共振波譜儀（瑞士Burker公司）；6550Q-TOF質(zhì)譜儀（美國(guó) Agilent Technologies）；WFH-203三用紫外分析儀（上海精科實(shí)業(yè)有限公司）；Sephadex LH-20（美國(guó)GE公司）；島津LC-6AD型半制備液相色譜儀（檢測(cè)器SPD-20A）；CAPCELL PAK C18MGⅡ半制備色譜柱（250 mm×10 mm，5 μm）；薄層色譜用硅膠GF254（青島海洋化工有限公司）；硅膠柱色譜（100～200、200～300目；青島海洋化工廠產(chǎn)品）；色譜甲醇，色譜乙腈（天津市科密歐化學(xué)試劑有限公司）；其他試劑均為分析純（天津市天力化學(xué)試劑有限公司）；人肺癌A549細(xì)胞、人結(jié)直腸癌HCT116細(xì)胞和SW620細(xì)胞均購(gòu)于中科院上海細(xì)胞庫(kù)。順鉑（批號(hào)B21M11L110000），源葉生物科技有限公司。

華中五味子根皮（長(zhǎng)勝七）于2019年9月采自陜西眉縣境內(nèi)，經(jīng)陜西中醫(yī)藥大學(xué)王薇教授鑒定為五味子科五味子屬植物華中五味子Rehd. et Wils的根皮。

2 提取與分離

干燥的華中五味子根皮（15.0 kg），用80%乙醇6倍量回流提取3次，每次1.5 h。減壓回收乙醇得浸膏，加水分散，用石油醚，醋酸乙酯依次萃取，得到醋酸乙酯部位450 g。醋酸乙酯部位（450 g）經(jīng)硅膠柱色譜分離，二氯甲烷-甲醇（1∶0～0∶1）梯度洗脫，得到14個(gè)組分（SSE1～14），SSE5（30 g）經(jīng)硅膠柱色譜，二氯甲烷-甲醇（1∶0～0∶1）梯度洗脫，得到7個(gè)組分（SSE5-1～5-7），SSE5-2經(jīng)葡聚糖凝膠Sephadex LH-20柱色譜，二氯甲烷-甲醇（1∶1）洗脫，合并相同流分后再經(jīng)半制備液相色譜，甲醇-水（69∶31）洗脫，得到化合物2（R＝22.3 min，12.3 mg）、3（R＝24.1 min，46.4 mg）、4（R＝30.5 min，9.2 mg）和5（R＝41.0 min，8.2 mg）。SSE5-4經(jīng)葡聚糖凝膠Sephadex LH-20柱色譜，二氯甲烷-甲醇（1∶1）洗脫，得到4個(gè)組分SSE5-4-1～SSE5-4-4，SSE5-4-3經(jīng)半制備液相色譜，乙腈-水（65∶35）洗脫得到化合物6（R＝13.4 min，10.5 mg）、7（R＝16.2 min，9.7 mg）、8（R＝20.9 min，11.4 mg）和9（R＝31.7 min，10.3 mg）。SSE5-4-4經(jīng)半制備液相色譜，甲醇-水（62∶38）洗脫，得到化合物1（R＝16.4 min，13.7 mg）。SSE6（8 g）經(jīng)硅膠柱色譜，二氯甲烷-甲醇（1∶0～0∶1）梯度洗脫，合并相同流分后，得到5個(gè)組分（SSE6-1～6-5），SSE6-2經(jīng)葡聚糖凝膠Sephadex LH-20柱色譜，二氯甲烷-甲醇（1∶1）洗脫，合并相同流分后，采用半制備液相色譜，甲醇-水（38∶62）洗脫，得到化合物10（R＝16.3 min，13.6 mg）和11（R＝21.1 min，8.6 mg）。

3 結(jié)構(gòu)鑒定

圖1 化合物1的化學(xué)結(jié)構(gòu)及主要的1H-1H COSY、HMBC和NOESY相關(guān)信號(hào)

圖2 化合物1的實(shí)驗(yàn)和計(jì)算ECD譜圖

化合物2：白色粉末（甲醇），ESI-MS/: 417 [M＋H]+；1H-NMR (400 MHz, CD3OD): 6.54 (1H, s, H-11), 6.46 (1H, s, H-4), 5.94 (2H, d,= 0.9 Hz, 12, 13-OCH2O), 5.91 (2H, m, 2, 3-OCH2O), 4.50 (1H, s, H-6), 3.84 (3H, s, -OCH3), 3.75 (3H, s, -OCH3), 2.46 (1H, dd,= 13.8, 9.7 Hz, H-9a), 2.02 (1H, d,= 13.8 Hz, H-9b), 1.81 (1H, m, H-8), 1.33 (3H, s, CH3-17), 1.09 (3H, d,= 7.2 Hz, CH3-18)；13C-NMR (100 MHz, CD3OD): 150.4 (C-12), 148.9 (C-3), 142.5 (C-1), 141.8 (C-14), 137.4 (C-13), 137.2 (C-2), 135.9 (C-5), 135.8 (C-10), 122.5 (C-16), 122.4 (C-15), 107.6 (C-4), 103.9 (C-11), 102.4 (12, 13-OCH2O), 102.07 (2, 3-OCH2O), 86.8 (C-6), 74.4 (C-7), 60.0 (-OCH3), 59.8 (-OCH3), 42.8 (C-8), 37.3 (C-9), 29.3 (C-17), 19.2 (C-18)。以上數(shù)據(jù)通過(guò)HSQC和HMBC譜圖進(jìn)行驗(yàn)證，與文獻(xiàn)報(bào)道基本一致[10]，該文獻(xiàn)中并沒(méi)有將此化合物命名，故將其命名為五味子醇丁。

表1 化合物1的1H-NMR (400 MHz, CD3OD) 和13C-NMR (100 MHz, CD3OD) 數(shù)據(jù)

Table 1 1H-NMR (400 MHz, CD3OD) and 13C-NMR (100 MHz, CD3OD) data of compound 1

碳位δHδC (type) 1—128.6 (C) 2—154.6 (C) 36.59 (1H, s)114.8 (CH) 4—134.0 (C) 57.16 (1H, s)122.2 (CH) 6—143.1 (C) 72.93 (1H, m)44.3 (CH) 81.75 (1H, m, H-8a)30.0 (CH2) 2.03 (1H, m, H-8b) 91.69 (1H, m, H-9a)24.3 (CH2) 1.73 (1H, m, H-9b) 102.73 (1H, m)33.8 (CH) 11—76.8 (C) 123.46 (2H, brs)68.9 (CH2) 131.13 (3H, s)24.3 (CH3) 141.27 (3H, d, J = 7.0 Hz)22.8 (CH3) 152.12 (3H, s)16.0 (CH3)

化合物3：白色粉末（甲醇），ESI-MS/: 417 [M＋H]+。1H-NMR (400 MHz, CDCl3): 6.51 (1H, s, H-11), 6.48 (1H, s, H-4), 6.00 (2H, d,= 1.2 Hz, 2, 3-OCH2O), 5.72 (1H, s, 12-OH), 3.92 (3H, s, 13-OCH3), 3.73 (6H, s, 1, 14-OCH3), 3.66 (1H, d,= 8.4 Hz, H-6), 3.02 (3H, s, 6-OCH3), 2.36 (1H, dd,= 13.0, 6.7 Hz, H-9a), 1.95 (1H, d,= 13.0 Hz, H-9b), 1.80 (1H, m, H-8), 1.65 (1H, m, H-7), 0.90 (3H, d,= 6.9 Hz, CH3-18), 0.85 (3H, d,= 7.0 Hz, CH3-17)；13C-NMR (100 MHz, CDCl3): 150.9 (C-1), 148.9 (C-12), 147.6 (C-3), 141.8 (C-14), 137.5 (C-10), 137.3 (C-13), 137.2 (C-2), 132.9 (C-5), 123.6 (C-16), 121.7 (C-15), 109.4 (C-11), 107.2 (C-4), 101.3 (2, 3- OCH2O), 90.5 (C-6), 60.9 (14-OCH3), 60.1 (13-OCH3), 59.8 (1-OCH3), 56.0 (6-OCH3), 38.8 (C-9), 36.9 (C-7), 36.8 (C-8), 18.3 (CH3-18), 17.1 (CH3-17)。以上數(shù)據(jù)與文獻(xiàn)報(bào)道基本一致[11]，故鑒定化合物3為12-demethylwuweilignan I。

化合物4：白色粉末（甲醇），ESI-MS/: 387 [M＋H]+。1H-NMR (400 MHz, CD3OD): 6.48 (1H, s, H-11), 6.41 (1H, s, H-4), 5.91 (2H, m, 2, 3-OCH2O), 3.86 (3H, s, 12-OCH3), 3.80 (3H, s, 13-OCH3), 3.79 (3H, s, 1-OCH3), 2.58 (1H, dd,= 13.4, 7.5 Hz, H-9a), 2.44 (1H, dd,= 13.4, 2.0 Hz, H-9b), 2.22 (1H, dd,= 13.2, 9.5 Hz, H-6a), 2.00 (1H, d,= 13.2 Hz, H-6b), 1.88 (1H, m, H-8), 1.78 (1H, m, H-7), 0.98 (3H, d,= 7.2 Hz, CH3-17), 0.74 (3H, d,= 7.1 Hz, CH3-18)；13C-NMR (100 MHz, CD3OD): 152.3 (C-12), 150.1 (C-3), 149.0 (C-1), 142.5 (C-14), 139.4 (C-10), 136.2 (C-5), 135.6 (C-13), 135.5 (C-2), 122.6 (C-15), 118.9 (C-16), 107.9 (C-4), 104.1 (C-11), 102.0 (2, 3-OCH2O), 61.1 (13-OCH3), 59.9 (1-OCH3), 56.3 (12-OCH3), 42.3 (C-8), 40.1 (C-6), 36.4 (C-9), 35.0 (C-7), 21.9 (CH3-17), 13.1 (CH3-18)。以上數(shù)據(jù)與文獻(xiàn)報(bào)道基本一致[12]，故鑒定化合物4為戈米辛M1。

化合物5：白色粉末（甲醇），ESI-MS/: 417 [M＋H]+。1H-NMR (400 MHz, CD3OD): 6.58 (1H, s, H-11), 6.56 (1H, s, H-4), 5.98 (2H, m, 2, 3-OCH2O), 4.22 (1H, d,= 9.2 Hz, H-6), 3.86 (3H, s, 1-OCH3), 3.85 (3H, s, 12-OCH3), 3.72 (3H, s, 13-OCH3), 3.69 (3H, s, 14-OCH3), 2.48 (1H, dd,= 12.8, 7.2 Hz, H-9a), 2.03 (1H, m, H-9b), 1.82 (1H, m, H-8), 1.50 (1H, m, H-7), 0.96 (3H, d,= 6.8 Hz, CH3-17), 0.88 (3H, d,= 7.0 Hz, CH3-18)；13C-NMR (100 MHz, CD3OD): 154.5 (C-12), 153.2 (C-14), 149.6 (C-3), 142.6 (C-1), 141.1 (C-13), 138.5 (C-10), 138.0 (C-2), 135.4 (C-5), 124.0 (C-16), 123.9 (C-15), 107.9 (C-11), 107.3 (C-4), 102.6 (2, 3-OCH2O), 81.5 (C-6), 61.3 (1-OCH3), 61.3 (14-OCH3), 59.9 (13-OCH3), 56.5 (12-OCH3), 40.2 (C-7), 38.9 (C-9), 34.7 (C-8), 18.5 (CH3-17), 16.9 (CH3-18)。以上數(shù)據(jù)與文獻(xiàn)報(bào)道基本一致[13]，故鑒定化合物5為isogomisin O。

化合物6：白色粉末（甲醇），ESI-MS/: 509 [M＋H]+。1H-NMR (400 MHz, CD3OD): 7.60～7.18 (5H, overlapped, -OBz), 6.75 (1H, s, H-4), 6.68 (1H, s, H-11), 5.98 (2H, m, 2, 3-OCH2O), 5.97 (1H, brs, H-6), 3.71 (3H, s, 1-OCH3), 2.80 (3H, s, 14-OCH3), 2.58 (1H, dd,= 13.9, 9.8 Hz, H-9a), 2.13 (1H, d,= 13.9 Hz, H-9b), 2.06 (1H, m, H-8), 1.25 (3H, s, CH3-17), 1.15 (3H, d,= 7.1 Hz, CH3-18)；13C-NMR (100 MHz, CD3OD): 166.7 (C-1′), 149.0 (C-3), 147.1 (C-13), 146.1 (C-14), 142.6 (C-1), 138.6 (C-2), 137.4 (C-12), 134.4 (C-2′), 134.1 (C-5′), 131.4 (C-10), 130.7 (C-5), 130.6 (C-4′, 6′), 129.4 (C-3′, 7′), 123.7 (C-16), 122.3 (C-15), 111.9 (C-11), 108.4 (C-4), 102.7 (2, 3-OCH2O), 85.7 (C-6), 73.7 (C-7), 60.2 (1-OCH3), 59.2 (14-OCH3), 43.9 (C-8), 37.0 (C-9), 28.9 (CH3-17), 19.2 (CH3-18)。以上數(shù)據(jù)與文獻(xiàn)報(bào)道基本一致[14]，故鑒定化合物6為schisphenlignan C。

化合物7：白色粉末（甲醇），ESI-MS/: 419 [M＋H]+。1H-NMR (400 MHz, CD3OD): 6.57 (1H, s, H-4), 6.41 (1H, s, H-11), 3.87 (3H, s, 2-OCH3), 3.84 (3H, s, 13-OCH3), 3.71 (3H, s, 14-OCH3), 3.42 (3H, s, 1-OCH3), 3.00 (3H, s, 6-OCH3), 3.66 (1H, m, H-6), 2.33 (1H, m, H-9a), 1.90 (1H, dd,= 12.9, 9.9 Hz, H-9b), 1.83 (1H, m, H-8), 1.62 (1H, m, H-7), 0.91 (3H, d,= 6.8 Hz, CH3-18), 0.86 (3H, d,= 6.9 Hz, CH3-17)；13C-NMR (100 MHz, CD3OD): 152.9 (C-1), 152.8 (C-3), 151.3 (C-14), 150.0 (C-12), 141.8 (C-2), 139.8 (C-13), 138.2 (C-10), 135.2 (C-5), 124.0 (C-16), 122.6 (C-15), 117.0 (C-4), 111.2 (C-11), 91.5 (C-6), 61.1 (1-OCH3), 61.0 (2-OCH3), 60.6 (13-OCH3), 60.3 (14-OCH3), 56.0 (6-OCH3), 39.7 (C-7), 38.5 (C-9), 38.3 (C-8), 18.3 (CH3-17), 16.8 (CH3-18)。以上數(shù)據(jù)與文獻(xiàn)報(bào)道基本一致[15]，故鑒定化合物7為marlignan F。

化合物8：白色粉末（甲醇），ESI-MS/: 403 [M＋H]+。1H-NMR (400 MHz, CD3OD): 6.93 (1H, s, H-4), 6.48 (1H, s, H-11), 5.93 (2H, brs, 12, 13-OCH2O), 4.42 (1H, d,= 1.5 Hz, H-6), 3.86 (3H, s, 1-OCH3), 3.75 (3H, s, 2-OCH3), 3.46 (3H, s, 3-OCH3), 2.15 (1H, m, H-7), 1.97 (2H, m, CH2-9), 1.87 (1H, m, H-8), 1.02 (3H, d,= 7.1 Hz, CH3-17), 0.66 (3H, d,= 7.0 Hz, CH3-18)；13C-NMR (100 MHz, CD3OD): 152.1 (C-1), 150.6 (C-3), 150.5 (C-12), 142.1 (C-2), 140.5 (C-14), 139.3 (C-5), 138.2 (C-10), 136.3 (C-13), 121.5 (C-16), 121.1 (C-15), 111.9 (C-4), 103.9 (C-11), 102.2 (12, 13-OCH2O), 74.0 (C-6), 61.2 (2-OCH3), 60.7 (1-OCH3), 59.9 (3-OCH3), 44.2 (C-7), 40.8 (C-8), 35.5 (C-9), 22.4 (CH3-18), 8.1 (CH3-17)。以上數(shù)據(jù)與文獻(xiàn)報(bào)道基本一致[16]，故鑒定化合物8為wilsonilignan A。

化合物9：淡綠色粉末（甲醇），ESI-MS/: 403 [M＋H]+。1H-NMR (400 MHz, CD3OD): 6.55 (1H, s, H-4), 6.43 (1H, s, H-11), 5.97 (2H, brs, OCH2O), 4.20 (1H, d,= 9.3 Hz, H-6), 3.86 (3H, s, 1-OCH3), 3.72 (3H, s, 2-OCH3), 3.69 (3H, s, 3-OCH3), 2.38 (1H, m, H-9a), 1.90 (1H, m, H-9b), 1.77 (1H, m, H-8), 1.54 (1H, m, H-7), 0.96 (3H, d,= 6.8 Hz, CH3-17), 0.86 (3H, d,= 6.9 Hz, CH3-18)；13C-NMR (100 MHz, CD3OD): 153.1 (C-1), 151.5 (C-3), 149.5 (C-12), 142.6 (C-2), 139.9 (C-14), 138.3 (C-10), 137.9 (C-13), 124.2 (C-16), 122.4 (C-5), 122.3 (C-15), 111.3 (C-4), 107.3 (C-11), 102.5 (12, 13-OCH2O), 81.5 (C-6), 61.1 (2-OCH3), 60.9 (1-OCH3), 59.9 (3-OCH3), 40.1 (C-7), 39.9 (C-9), 39.1 (C-8), 18.6 (CH3-18), 16.2 (CH3-17)。以上數(shù)據(jù)與文獻(xiàn)中化合物數(shù)據(jù)基本一致[17]，故鑒定化合物9為sphenlignan A。

化合物10：白色粉末（甲醇），ESI-MS/: 389 [M＋H]+。1H-NMR (400 MHz, CD3OD): 6.53 (1H, s, H-4), 6.43 (1H, s, H-11), 3.86 (3H, s, 12-OCH3), 3.82 (3H, s, 2-OCH3), 3.79 (3H, s, 14-OCH3), 3.54 (3H, s, 1-OCH3), 2.60 (1H, dd,= 13.5, 7.6 Hz, H-6a), 2.45 (1H, dd,= 13.5, 1.9 Hz, H-6b), 2.21 (1H, dd,= 13.0, 9.4 Hz, H-9a), 1.97 (1H, d,= 13.0 Hz, H-9b), 1.74 (1H, m, H-8), 1.88 (1H, m, H-7), 0.98 (3H, d,= 7.1 Hz, CH3-18), 0.74 (3H, d,= 7.1 Hz, CH3-17)；13C-NMR (100 MHz, CD3OD): 152.4 (C-12), 152.3 (C-14), 151.4 (C-1), 148.9 (C-3), 141.3 (C-2), 139.7 (C-13), 135.7 (C-5), 135.6 (C-10), 122.0 (C-16), 119.1 (C-15), 112.4 (C-4), 108.0 (C-11), 61.1 (14-OCH3), 61.0 (2-OCH3), 60.8 (1-OCH3), 56.2 (12-OCH3), 42.6 (C-8), 40.1 (C-6), 36.2 (C-9), 35.2 (C-7), 22.1 (C-17), 13.0 (C-18)。以上數(shù)據(jù)與文獻(xiàn)報(bào)道基本一致[18]，故鑒定化合物10為schinegllignan A。

化合物11：白色粉末（甲醇），ESI-MS/: 389 [M＋H]+。1H-NMR (400 MHz, CD3OD): 6.55 (1H, s, H-4), 6.41 (1H, s, H-11), 3.87 (3H, s, 12-OCH3), 3.84 (3H, s, 13-OCH3), 3.80 (3H, s, 2-OCH3), 3.55 (3H, s, 14-OCH3), 2.52 (1H, dd,= 13.4, 7.5 Hz, H-6a), 2.41 (1H, dd,= 13.4, 2.0 Hz, H-6b), 2.24 (1H, dd,= 13.1, 9.4 Hz, H-9a), 2.04 (1H, d,= 13.1 Hz, H-9b), 1.86 (1H, m, H-7), 1.43 (1H, m, H-8), 1.00 (3H, d,= 7.1 Hz, CH3-18), 0.73 (3H, d,= 7.1 Hz, CH3-17)；13C-NMR (100 MHz, CD3OD): 153.6 (C-14), 152.7 (C-12), 150.1 (C-3), 148.7 (C-1), 140.9 (C-13), 140.0 (C-2), 135.9 (C-10), 135.3 (C-5), 123.1 (C-16), 118.1 (C-15), 115.7 (C-4), 104.4 (C-11), 61.2 (13-OCH3), 61.1 (14-OCH3), 60.8 (2-OCH3), 56.2 (12-OCH3), 42.4 (C-8), 39.8 (C-6), 36.5 (C-9), 35.3 (C-7), 22.1 (C-17), 13.0 (C-18)。以上數(shù)據(jù)與文獻(xiàn)報(bào)道基本一致[18]，故鑒定化合物11為schinegllignan B。

4 細(xì)胞毒活性測(cè)試

采用MTT法測(cè)定化合物1～11對(duì)A549細(xì)胞、HCT116細(xì)胞和SW620細(xì)胞的細(xì)胞毒活性，順鉑為陽(yáng)性對(duì)照，實(shí)驗(yàn)所用儀器材料以及實(shí)驗(yàn)過(guò)程均與本課題組前期報(bào)道一致[19-20]。實(shí)驗(yàn)結(jié)果（表2）表明化合物2～5和7對(duì)A549、HCT116、SW620細(xì)胞株具有細(xì)胞毒活性，IC50值為12.1～38.7 μmol/L，其余化合物的IC50值均大于50 μmol/L。

5 討論

從華中五味子根皮中分離鑒定了11個(gè)化合物，包括1個(gè)新的倍半萜（1），10個(gè)聯(lián)苯環(huán)辛烯型木脂素（2～11），其中化合物2為新的天然產(chǎn)物，化合物3～5、7、8、10和11為首次從該植物中分離得到，進(jìn)一步豐富了華中五味子的化學(xué)成分。細(xì)胞毒活性實(shí)驗(yàn)顯示化合物2～5和7具有細(xì)胞毒活性，但本文僅測(cè)定了11個(gè)化合物的體外細(xì)胞毒活性，關(guān)于這些化合物的其他活性有待于進(jìn)一步研究。

表2 化合物1～11對(duì)3種腫瘤細(xì)胞的細(xì)胞毒活性

Table 2 Cytotoxic activities of 1－11 in three cancer cell lines

化合物IC50/(μmol·L?1)A549HCT116SW620 1＞50＞50＞50 231.7±2.4＞50＞50 317.4±1.523.6±0.625.2±1.2 413.3±1.412.1±0.518.8±0.7 516.9±0.214.4±0.618.4±0.4 6＞50＞50＞50 738.7±0.5＞50＞50 8＞50＞50＞50 9＞50＞50＞50 10＞50＞50＞50 11＞50＞50＞50 順鉑32.1±1.343.5±3.332.5±3.4

利益沖突 所有作者均聲明不存在利益沖突

[1] 毛水龍. 秦嶺七藥 [M]. 西安: 西安交通大學(xué)出版社, 2011: 68-69.

[2] 劉媛媛, 黃仕其, 李玉澤, 等. 五味子屬植物木脂素類化學(xué)成分及其藥理作用研究進(jìn)展[J]. 中草藥, 2022, 53(6): 1903-1918.

[3] Huang S Q, Zhang D D, Li Y Z,.: A comprehensive review of its botany, phytochemistry, pharmacology, and clinical applications [J]., 2021, 49(7): 1577-1622.

[4] Huang S Q, Liu Y Y, Li Y Z,. Dibenzocyclooctadiene lignans from the root bark of[J]., 2021, 45: 137-141.

[5] 黃澤豪, 秦路平. 華中五味子藤莖的化學(xué)成分研究 [J]. 中草藥, 2016, 47(19): 3374-3378.

[6] 金銀萍, 焉石, 劉俊霞, 等. 五味子科植物中降三萜類成分及其藥理作用研究進(jìn)展 [J]. 中草藥, 2014, 45(11): 1643-1650.

[7] Salmoun M, Braekman J C, Ranarivelo Y,. New calamenene sesquiterpenes from[J]., 2007, 21(2): 111-120.

[8] Zhang D D, Shi Y H, Shi S S,. Isatisindigoticanine A, a novel indole alkaloid with an unpresented carbon skeleton from the roots of[J]., 2021, 35(8): 1249-1255.

[9] Zhang D D, Sun Y, Chen Z Q,. Bisindole alkaloids with nitric oxide inhibitory activities from an alcohol extract of theindigotica roots [J]., 2020, 146:104654.

[10] Ikeya Y, Taguchi H, Yosioka I. The constituents ofBaill. XII. Isolation and structure of a new lignan, gomisin R, the absolute structure of wuweizisu C and isolation of schisantherin D [J]., 1982, 30(9): 3207-3211.

[11] Luo X, Pu J X, Fan P,. Four new dibenzocyclooctadiene lignans from[J]., 2011, 9(3): 167-172.

[12] Ikeya Y, Taguchi H, Yosioka I. The Constituents ofBaill. X. The Structures of.GAMMA.-schizandrin and four new lignans, (?)-gomisins L1 and L2, (+)-gomisin M1 and (+)-gomisin M2 [J]., 1982, 30(1): 132-139.

[13] Ikeya Y, Ookawa N, Taguchi H,. The constituents ofBaill. XI. The structures of three new lignans, angeloylgomisin O, and angeloyl- and benzoylisogomisin O [J]., 1982, 30(9): 3202-3206.

[14] Liang C Q, Hu J, Shi Y M,. Schisphenlignans A-E: Five new dibenzocyclooctadiene lignans from[J]., 2013, 61(1): 96-100.

[15] Yang G Y, Li Y K, Wang R R,. Dibenzocyclooctadiene lignans fromand their anti-HIV-1 activities [J]., 2010, 73(5): 915-919.

[16] Yang G Y, Li Y K, Wang R R,. Dibenzocyclooctadiene lignans from the fruits ofand their anti-HIV-1 activities [J]., 2010, 12(6): 470-476.

[17] 賀飛. 三種藥用植物的化學(xué)成分和生物活性研究 [D]. 北京: 中國(guó)科學(xué)院昆明植物研究所, 2009.

[18] Duan Y X, Cao J L, Wen R R,. Dibenzocyclooctadiene lignans fromand their anti-HIV-1 activities [J]., 2011, 13(7): 592-598.

[19] Sun Y, Ding C, Wang F R,. Pregnane alkaloids with BRD4 inhibitory and cytotoxic activities from[J]., 2021, 45: 63-67.

[20] 張東東, 樊浩, 孫玉, 等. 纈草中1個(gè)新的單環(huán)氧木脂素 [J]. 中草藥, 2022, 53(1): 25-30.

A new calamenene sesquiterpenoid from root bark of

ZHANG Dong-dong, HUANG Shi-qi, LIU Yuan-yuan, SUN Yu, LI Yu-ze, ZHANG Hua-wei, DENG Chong, SONG Xiao-mei, WANG Wei

School of Pharmacy, Shaanxi University of Chinese Medicine, Xianyang 712046, China

To study the chemical constituents from the root bark ofand their cytotoxic activities.The compounds were isolated and purified by silica, sephadexLH-20 and semi preparative-HPLC and the chemical structures were identified by NMR, MS and ECD data analysis.Eleven compounds were isolated and deduced as: (7,10,11)-2,11,12-trihydroxycalamenene (1), schisandrol D (2), 12-demethylwuweilignan I (3), gomisin M1 (4), isogomisin O (5), schisphenlignan C (6), marlignan F (7), wilsonilignan A (8), sphenlignan A (9), schinegllignan A (10), schinegllignan B (11). Compounds 2—5 and 7 showed cytotoxic activity against A549, HCT116 and SW620 cell lines with IC50values ranging from 12.1 to 38.7 μmol/L.Compound 1 is a new sesquiterpene, named as schisphensesquiterpene A, and compound 2 is a new natural product. Compounds 3—5, 7, 8, 10—11 were isolated from theplant for the first time, and compounds 2—5 and 7 exhibited cytotoxic activities.

Rehd. et Wils; schisphensesquiterpene A; schisandrol D;gomisin M1; isogomisin O; lignans; cytotoxity

R284.1

A

0253 - 2670(2022)14 - 4270 - 06

10.7501/j.issn.0253-2670.2022.14.006

2022-01-21

國(guó)家自然科學(xué)基金項(xiàng)目（82174111）；國(guó)家自然科學(xué)基金項(xiàng)目（82104368）；陜西中醫(yī)藥大學(xué)學(xué)科創(chuàng)新團(tuán)隊(duì)項(xiàng)目（2019-YL12）

張東東，男，博士，講師，主要從事中藥藥效物質(zhì)基礎(chǔ)研究。E-mail: zhangnatprod@163.com

王 薇，女，博士，教授，主要從事中藥藥效物質(zhì)基礎(chǔ)及中藥炮制研究。E-mail: 2051003@sntcm.edu.cn

[責(zé)任編輯 王文倩]