國產(chǎn)綠奇楠沉香的化學(xué)成分研究

李薇, 梅文莉, 董文化, 蔡彩虹, 蓋翠娟, 戴好富

?

國產(chǎn)綠奇楠沉香的化學(xué)成分研究

李薇, 梅文莉, 董文化, 蔡彩虹, 蓋翠娟, 戴好富*

(中國熱帶農(nóng)業(yè)科學(xué)院熱帶生物技術(shù)研究所，農(nóng)業(yè)部熱帶作物生物學(xué)與遺傳資源利用重點(diǎn)實(shí)驗(yàn)室，海南省沉香工程技術(shù)研究中心，海南省黎藥資源天然產(chǎn)物研究與利用重點(diǎn)實(shí)驗(yàn)室，海口 571101)

為了解國產(chǎn)綠‘奇楠’沉香的化學(xué)成分，采用色譜和波譜方法從其乙醚和乙醇提取物中分離鑒定了7個(gè)化合物，分別鑒定為順式-7-羥基菖蒲烯 (1), (5,6,7,8)-2-(苯乙基)-6,7,8-三羥基-5,6,7,8-四氫-5-[2-(2-苯乙基)色酮基-6-氧代]色酮 (2), 1-羥基-1,5-二苯基戊-3-酮 (3), 丁香樹脂酚葡萄糖苷 (4), (3)-齊墩果-12-烯-3,23-二醇(5),-谷甾醇 (6)和棕櫚酸--單甘油酯 (7)?；衔?、3~5和7均為首次從沉香中分離得到，其中化合物1表現(xiàn)出非常甜的芳香氣味。乙酰膽堿酯酶體外抑制活性測試結(jié)果表明，50mol L–1的化合物1對(duì)乙酰膽堿酯酶抑制率為(49.9±1.4)%。

奇楠；沉香；化學(xué)成分；乙酰膽堿酯酶；抑制活性

‘奇楠’又名伽楠、伽羅、琪南、伽沉等，英文名有Kanankoh、Kyara、Chi-Nan、Qi-Nan等，因其神秘優(yōu)雅的香味被認(rèn)為是具有最上等品質(zhì)和最珍貴價(jià)值的沉香，且在市場上價(jià)格也最為昂貴[1–2]?！骈谖锢硖卣鞣矫嬷饕谄浞枷銡馕逗屯庥^差別而與普通沉香相區(qū)別[3–4]，并根據(jù)其外表和斷面的顏色可分為綠奇、紫奇、黑奇和黃奇等[2,5]。據(jù)記載奇楠外表油潤光滑，油性重，以指甲刻之，如錐畫沙，油隨即溢出，用刀刮削，能捻捏成丸、成餅，能散發(fā)耐久的幽香，味微苦麻辣，嚼之粘牙,燃之出油，而普通沉香質(zhì)堅(jiān)，雕剔之，如刀刮竹[5]。在化學(xué)特征方面，根據(jù)本研究組前期對(duì)‘奇楠’沉香化學(xué)成分的分析和分離研究，發(fā)現(xiàn)‘奇楠’沉香中有較多母核無取代的2-(2-苯乙基)色酮類化合物，而這種類型的2-(2-苯乙基)色酮在普通沉香中幾乎沒有過報(bào)道[6–7]。同時(shí)，在前期研究中我們還發(fā)現(xiàn)，‘奇楠’沉香中有兩個(gè)含量非常大的2-(2-苯乙基)色酮類成分，即2-(2-苯乙基)色酮和2-[2-(4-甲氧基苯)乙基]色酮，其二者相對(duì)含量之和分別占到了研究的4種‘奇楠’沉香(海南產(chǎn)的白奇、紫奇、綠奇和越南產(chǎn)的綠奇)乙醚提取物的66.47%、82.09%、84.71%和71.98%，因此，也可以將上述兩個(gè)2-(2-苯乙基)色酮在乙醚提取物中的相對(duì)含量之和作為區(qū)分‘奇楠’沉香與普通沉香的一項(xiàng)重要指標(biāo)[8]。本研究采用的沉香樣品為產(chǎn)自于我國廣東省的綠‘奇楠’沉香，其基源植物白木香[(Lour.) Gilg]是我國特有樹種，主要分布在廣東、廣西、福建、海南和臺(tái)灣等地[9]。

本研究組在前期研究中，已從國產(chǎn)綠‘奇楠’沉香中分離鑒定出一系列新的2-(2-苯乙基)色酮和倍半萜類化合物[6,10?13]，為了進(jìn)一步完善對(duì)‘奇楠’沉香的化學(xué)成分研究，本次從國產(chǎn)綠‘奇楠’沉香乙醚和乙醇提取物中共分離鑒定出7個(gè)化合物，其中化合物1、3~5和7均為首次從沉香中分離得到的化合物，且本研究還發(fā)現(xiàn)化合物1在體外對(duì)乙酰膽堿酯酶有一定的抑制作用。本文報(bào)道了化合物1~7的分離方法、結(jié)構(gòu)鑒定和乙酰膽堿酯酶抑制活性測試結(jié)果。

1 材料和方法

1.1 材料

綠‘奇楠’沉香，表面可觀測到淺褐色細(xì)密樹脂條帶均勻分布其上(圖1)，質(zhì)地較普通沉香更軟，不點(diǎn)燃即可聞到有明顯的沁人心脾的香氣，被稱為金絲軟綠奇，于2011年8月由沉香收藏家張曉武先生及官茂有先生惠贈(zèng)，由中國熱帶農(nóng)業(yè)科學(xué)院熱帶生物技術(shù)研究所戴好富研究員與中國醫(yī)學(xué)科學(xué)院藥用植物研究所海南分所鄭希龍博士共同鑒定其源植物為白木香[(Lour.) Gilg]，憑證標(biāo)本(QN 20110830)存放于中國熱帶農(nóng)業(yè)科學(xué)院熱帶生物技術(shù)研究所。

圖1 綠奇楠沉香樣品(編號(hào): QN 20110830)

1.2 儀器和試劑

Buker amazon SL質(zhì)譜儀；Bruker AV-500型超導(dǎo)核磁儀，TMS為內(nèi)標(biāo)；XY-JH-21BC超凈工作臺(tái)(上海昕儀儀器儀表有限公司)；Multiskan FC酶標(biāo)儀(塞默飛世爾上海儀器有限公司)；旋轉(zhuǎn)蒸發(fā)儀(德國海道夫公司)；Delta 320-S pH計(jì)(梅特勒-托利多儀器上海有限公司)；柱色譜硅膠(200~300目，60~80目)和薄層層析硅膠板G (青島海洋化工廠), Sephadex LH-20和RP-18填料柱(Merck公司)，提取和分離所用試劑均為重蒸工業(yè)試劑。S-硫代乙酰膽堿，5,5?-二硫-雙-2-硝基苯甲酸(DTNB, Ellman試劑)，乙酰膽堿酯酶，他克林均購自Sigma Chemical。

1.3 提取和分離

綠奇楠沉香(干重180.2 g)粉碎后，以乙醚超聲提取4次，每次超聲0.5 h，靜置5 h。合并濾液, 減壓濃縮得油狀乙醚提取物19.3 g，得率為10.7%。將乙醚提取物留樣20.0 mg，取5.0 mg用作GC-MS分析后，再以硅膠柱色譜分離，以石油醚-乙酸乙酯梯度(體積比1∶0~0∶1) 洗脫，得到11個(gè)流分(D1~ D11)。乙醚提取后，剩余濾渣用乙醇加熱回流提取3次，減壓濃縮，得到乙醇提取物15.8 g，以減壓硅膠柱(C∶M=1∶0~0∶1)色譜分離，得到11個(gè)流分(E1~E11)。

D2 (與E1合并后1.7 g)以甲醇溶解，不溶解部分(400.0 mg)反復(fù)柱層析(P∶E=20∶1)得到化合物6 (1.0 mg)。甲醇溶解部分經(jīng)Sephadex LH-20柱色譜分離，甲醇洗脫得到D2-1和D2-2。D2-2 (400.0 mg)以硅膠柱色譜分離，石油醚-乙酸乙酯梯度(80∶1?10∶1，/, 下同)洗脫，得到化合物1 (1.3 mg)。D3 (3.8 g)以Sephadex LH-20柱色譜分離，甲醇洗脫得到D3-1 (2.3 g)和D3-2 (1.5 g)。D3-1 (2.3 g)以反相ODS柱色譜分離，甲醇-水梯度(40∶60~ 100∶0)洗脫得到10個(gè)流分(D3-1-1~D3-1-10)。D3- 1-5 (43.0 mg)以Sephadex LH-20柱色譜分離，甲醇洗脫后得到D3-1-5-1和D3-1-5-2；D3-1-5-2 (7.9 mg)以硅膠柱色譜分離，石油醚-氯仿(3∶7)洗脫得到化合物3 (5.8 mg)。D6 (1.6 g)以甲醇溶解，不溶部分(32.0 mg)以硅膠柱色譜分離，氯仿-甲醇(100∶1) 洗脫得到化合物7 (11.1 mg)。D-8 (1.7 g)以反相ODS柱色譜分離，甲醇-水梯度(30∶70~100∶0)洗脫得到D-8-1~D-8-4，D-8-4以葡聚糖凝膠色譜分離，甲醇洗脫得到化合物5 (4.7 mg)。D11 (1.2 g)以反相ODS柱色譜分離，甲醇-水梯度(3∶7?1∶0) 洗脫，得到D11-1~D11-70, D11-(62+63) (30.0 mg)以硅膠柱色譜分離，氯仿-甲醇(25∶1)洗脫得到化合物2 (5.0 mg)。E8 (2.6 g)經(jīng)ODS(甲醇-水=5∶5~1∶0)梯度洗脫，得到E8-1~E8-4。E8-4 (44.1 mg)經(jīng)Sephadex LH-20 (甲醇)再經(jīng)硅膠柱(C∶M=20∶1) 色譜分離，得到化合物4 (1.7 mg)?；衔锝Y(jié)構(gòu)見圖2。

圖2 化合物1~7的結(jié)構(gòu)

1.4 乙酰膽堿酯酶抑制活性的測試方法

化合物的乙酰膽堿酯酶抑制活性測定參照Ellman法[14]并略有改進(jìn)。化合物溶解于DMSO中, 含有磷酸緩沖液(pH=8.0)、測試化合物(50mol L–1)和乙酰膽堿酯酶(0.02 U mL?1)的最終反應(yīng)體系(200L),先在30℃孵育20 min，于405 nm紫外光下測兩次背景值, 加入20L的DTNB (2.48 mg mL?1)和20L的硫代乙酰膽堿(1.81 mg mL?1)開始反應(yīng)，將反應(yīng)體系于405 nm紫外波長下監(jiān)測1 h。他克林用作陽性對(duì)照，反應(yīng)終濃度為0.08g mL?1；DMSO用作陰性對(duì)照，反應(yīng)終濃度為0.1%。每個(gè)樣品重復(fù)3次實(shí)驗(yàn)。選擇陰性對(duì)照組吸光值平均約為1時(shí)的樣品吸光值，計(jì)算化合物吸光值平均值(化合物測定值-背景值)，化合物的抑制率(%)=(–)/×100, 式中,為陰性對(duì)照組的平均吸光值；為化合物組的平均吸光值。

2 結(jié)果和分析

2.1 結(jié)構(gòu)鑒定

順式-7-羥基菖蒲烯(-7-hydroxycalamenene, 1) 芳香味無色油狀；遇體積分?jǐn)?shù)5%的硫酸顯藍(lán)色。分子式C15H22O。ESI-MS: 241.1 (100) [M + Na]+。1H NMR (CDCl3, 500 MHz):6.88 (1H, s, H-5), 6.64 (1H, s, H-8), 2.80 (1H, m, H-1), 2.54 (1H, m, H-4), 2.20 (1H, m, H-12), 2.16 (3H, s, H-15), 1.77 (1H, m, H-2a), 1.68 (2H, m, H-3), 1.63 (1H, m, H-2b), 1.21 (3H, d,= 7.1 Hz, H-11), 1.02 (3H, d,= 6.9 Hz, H-14), 0.75 (3H, d,= 6.8 Hz, H-13)；13C NMR (CDCl3, 125 MHz):32.2 (C-1), 29.0 (C-2), 19.3 (C-3), 43.5 (C-4), 131.1 (C-5), 121.1 (C-6), 151.6 (C-7), 114.0 (C-8), 139.0 (C-9), 135.4 (C-10), 23.6 (C-11), 31.1 (C-12), 17.4 (C-13), 21.5 (C-14), 15.5 (C-15)。以上數(shù)據(jù)與文獻(xiàn)[15]一致，確定化合物1為順式-7-羥基菖蒲烯。

(5,6,7,8)-2-(苯乙基)-6,7,8-三羥基-5,6,7,8-四氫-5-[2-(2-苯乙基)色酮基-6-氧代]色酮(AH14) [(5,6,7,8)-2-(phenylethyl)-6,7,8-trihydroxy-5,6,7,8-tetrahydro-5-[2-(2-phenylethyl)chromonyl-6-oxy] chromone, 2] 白色粉末；遇碘顯黃色，遇體積分?jǐn)?shù)5%的硫酸顯淺紅色。分子式C34H30O8。ESI-MS: 589.4 (100) [M+Na]+。1H NMR (CDCl3, 500 MHz)H: Unit A: 7.29 (2H, m, H-2″, H-6″), 6.96 (2H, d,= 7.0 Hz, H-3″, H-5″), 7.20 (1H, m, H-4″), 6.11 (1H, s, H-3), 5.35(1H, dd,= 7.4, 1.3 Hz, H-5), 4.86 (1H, dd,= 7.5, 1.3 Hz, H-8),4.10 (1H, dd,= 10.2, 7.4 Hz, H-6), 3.91 (1H, dd,= 10.2, 7.5 Hz, H-7), 2.68 (4H, m, H-7″, H-8″), Unit B: 7.20 (5H, m, H-2?~6?), 7.93 (1H, d,= 1.8 Hz, H-5′), 7.42 (1H, d,= 9.2 Hz, H-8′), 7.32 (1H, dd,= 9.2, 1.8 Hz, H-7′),6.15 (1H, s, H-3′), 3.05 (2H, m, H-7?), 2.94 (2H, m, H-8?);13C NMR (CDCl3, 125 MHz)C: Unit A: 169.4(C-2), 113.8 (C-3), 180.6 (C-4), 78.4 (C-5), 72.5 (C-6), 74.0 (C-7), 69.9 (C-8), 157.9 (C-9), 121.1 (C-10), 139.2 (C-1″), 128.8 (C-2″, C-6″), 128.2 (C-3″, C-5″), 126.8 (C-4″), 32.6 (C-7″), 35.4 (C-8″), Unit B:168.8 (C-2′), 109.7 (C-3′), 178.0 (C-4′), 109.6 (C-5′), 156.5 (C-6′), 124.7 (C-7′), 119.7 (C-8′), 152.1 (C-9′), 124.4 (C-10′), 139.8 (C-1?), 128.8 (C-2?, C-6?), 128.4 (C-3?, C-5?), 126.8 (C-4?), 33.1 (C-7?), 36.2 (C-8?)。上述數(shù)據(jù)與文獻(xiàn)[16]報(bào)道的AH14的數(shù)據(jù)一致，確定化合物2為(5,6,7,8)-2-(苯乙基)-6,7,8-三羥基-5,6,7,8-四氫-5-[2-(2-苯乙基)色酮基-6-氧代]色酮。

1-羥基-1,5-二苯基戊-3-酮 (1-hydroxy-1,5-di- phenylpentan-3-one, 3) 無色油狀；遇體積分?jǐn)?shù)5%的硫酸淺黃色。分子式C17H18O2。ESI-MS: 277.1 (100) [M+Na]+。1H NMR (CDCl3, 500 MHz):7.35 (4H, m, H-2″, H-6″, H-3￠, H-5￠), 7.29 (3H, m, H-2￠, H-4￠, H-6￠), 7.21 (1H, m, H-4″), 7.17 (2H, m, H-3″, H-5″), 5.15 (1H, dd,= 3.0, 9.3 Hz, H-1), 2.91 (2H, t,= 7.6 Hz, H-5), 2.85 (1H, dd,= 9.3, 17.3 Hz, H-2a), 2.78 (2H, m, H-4), 2.74 (1H, dd,= 3.0, 17.3 Hz, H-2b);13C NMR (CDCl3, 125 MHz):70.1 (C-1), 51.4 (C-2), 210.4 (C-3), 45.2 (C-4), 29.6 (C-5), 140.8 (C-1￠), 128.4 (C-2￠, C-6￠), 125.7 (C-3￠, C-5￠), 127.8 (C-4￠), 142.8 (C-1￠￠), 128.7 (C-2″, C-3″, C-5″, C-6″), 126.4 (C-4″)。以上數(shù)據(jù)與文獻(xiàn)[17]一致，確定化合物3為1-羥基-1,5-二苯基戊-3-酮。

丁香樹脂酚葡萄糖苷(syringaresinol-glyco- side, 4) 淡黃色粉末(MeOD); ESI-MS: 603 [M + Na]+, C28H36O13;1H NMR (MeOD, 500 MHz):6.71 (2H, s, H-2″, H-6″), 6.65 (2H, s, H-2′, H-6′), 4.90 (1H, d,= 7.5 Hz, G-H-1), 4.76 (1H, d,= 4.0 Hz, H-6), 4.71 (1H, d,= 4.3 Hz, H-2), 3.85 (6H, s, 3″, 5″-OCH3), 3.83 (6H, s, 3′, 5′-OCH3);13C NMR (MeOD, 125 MHz):154.7 (C-3″, C-5″), 149.7 (C-3', C-5'), 139.8 (C-4″), 136.5 (C-4′), 135.8 (C-1″), 133.4 (C-1′), 105.6 (G-C-1), 105.2 (C-2″, C-6″), 104.8 (C-2′, C-6′), 87.8 (C-2), 87.5 (C-6), 78.6 (G-C-5), 78.1 (G-C-3), 75.9 (G-C-2), 73.2 (C-4), 73.2 (C-8), 71.6 (G-C-4), 62.9 (G-C-6), 57.4 (2×OCH3), 57.1 (2× OCH3), 56.1 (C-1), 55.8 (C-5)。以上數(shù)據(jù)與文獻(xiàn)[18?19]對(duì)照，鑒定該化合物為丁香樹脂酚葡萄糖苷。

(3)-齊墩果-12-烯-3,23-二醇 [(3)-olean-12- ene-3,23-diol, 5] 白色無定型固體；遇體積分?jǐn)?shù)5%的硫酸顯紫色。分子式C30H50O2。ESI-MS: 465 (100) [M+Na]+。1H NMR (CDCl3, 500 MHz):5.18 (1H, t,= 4.8 Hz, H-12), 3.68 (1H, d,= 9.8 Hz, H-23a), 3.52 (1H, t,= 7.6 Hz, H-3), 3.42 (1H, d,= 9.8 Hz, H-23b), 2.24 (1H, m, H-18), 2.01 (1H, m, H-15a), 1.93 (2H, m, H-11), 1.78 (1H, m, H-16a), 1.73 (1H, m, H-1a), 1.70 (2H, m, H-2), 1.62 (1H, m, H-9), 1.60 (1H, m, H-7a), 1.52 (1H, m, H-6a), 1.50 (1H, m, H-22a), 1.42 (1H, m, H-6b), 1.40 (1H, m, H-21a), 1.37 (1H, m, H-7b), 1.25 (H, m, H-22b), 1.14 (3H, s, H-27), 1.08 (1H, m, H-21b), 1.03 (1H, m, H-19b), 1.02 (2H, m, H-1b, H-16b), 1.00 (3H, s, H-25), 0.98 (3H, s, H-26), 0.92 (3H, s, H-24), 0.88 (6H, s, H-29, H-30), 0.86 (1H, m, H-5), 0.84 (3H, s, H-28), 0.79 (1H, s, H-15b);13C NMR (CDCl3, 125 MHz):37.8 (C-1), 27.4 (C-2), 77.4 (C-3), 41.5 (C-4), 49.1 (C-5), 18.0 (C-6), 32.4 (C-7), 38.9 (C-8), 47.3 (C-9), 36.6 (C-10), 23.1 (C-11), 121.8 (C-12), 143.8 (C-13), 41.3 (C-14), 25.7 (C-15), 23.1 (C-16), 46.2 (C-17), 41.1 (C-18), 45.8 (C-19), 30.4 (C-20), 33.6 (C-21), 32.1 (C-22), 70.2 (C-23), 11.2 (C-24), 15.3 (C-25), 16.5 (C-26), 25.5 (C-27), 25.5 (C-28), 32.7 (C-29), 23.1 (C-30)。以上數(shù)據(jù)與文獻(xiàn)[20]一致，確定化合物5為(3)-齊墩果-12-烯-3,23-二醇。

-谷甾醇 (-sitosterol, 6) 白色粉末，遇體積分?jǐn)?shù)5%的硫酸顯紫色。分子式為C29H50O。ESI- MS: 414 [M]+。1H NMR (CDCl3, 500 MHz):5.36 (1H, br s, H-6), 3.51 (1H, m, H-3), 1.01 (3H, s, H-19), 0.92 (3H, d,= 6.4 Hz, H-21), 0.84 (3H, d,= 7.8 Hz, H-29), 0.84 (3H, d,= 6.8 Hz, H-26), 0.80 (3H, d,= 6.9 Hz, H-27), 0.69 (3H, s, H-18);13C NMR (CDCl3, 125 MHz):37.4 (C-1), 31.8 (C-2), 72.0 (C-3), 42.4 (C-4), 140.9 (C-5), 121.9 (C-6), 32.0 (C-7), 31.8 (C-8), 50.3 (C-9), 36.6 (C-10), 21.2 (C-11), 39.8 (C-12), 42.4 (C-13), 56.9 (C-14), 24.4 (C-15), 28.4 (C-16), 56.2 (C-17), 12.1 (C-18), 19.5 (C-19), 36.3 (C-20), 18.9 (C-21), 34.1 (C-22), 26.2 (C-23), 46.0 (C-24), 29.3 (C-25), 20.0 (C-26), 19.2 (C-27), 23.2 (C-28), 12.0 (C-29)。以上數(shù)據(jù)與文獻(xiàn)[21]報(bào)道的一致，確定化合物6為-谷甾醇。

棕櫚酸--單甘油酯 (hexadecanoic acid, 2,3- dihydroxypropyl ester, 7) 白色粉末，遇體積分?jǐn)?shù)5%的硫酸顯淺黃色。分子式為C19H38O4。ESI-MS:329.3 (10)[M-H]-。1H NMR (CDCl3, 500 MHz):4.13 (1H, dd,= 4.6, 11.4 Hz), 4.06 (1H, dd,= 6.2, 11.6 Hz), 3.83 (1H, dt,= 5.4, 10.7 Hz), 3.57 (1H, dd,= 5.0, 11.4 Hz), 3.53 (1H, dd,= 5.8, 11.4 Hz), 2.33 (2H, d,= 7.6 Hz), 1.59 (2H, dd,= 7.0, 14.4 Hz), 1.26 (24H, m), 0.86 (3H, t,= 6.9 Hz, H-16);13C NMR (CDCl3, 125 MHz):173.9 (C-1), 69.2 (C-1￠), 64.6 (C-2￠), 62.3 (C-3￠), 33.3 (C-2), 31.2 (C-14), 28.4~28.9 (C-4-14), 24.2 (C-3), 21.9 (C-15), 13.0 (C- 16)。以上數(shù)據(jù)與文獻(xiàn)[22]一致，確定化合物7為棕櫚酸--單甘油酯。

2.2 乙酰膽堿酯酶抑制活性

參照Ellman法[14]并略有改進(jìn)后，對(duì)國產(chǎn)綠奇楠沉香中化合物1~7的乙酰膽堿酯酶抑制活性進(jìn)行測定，結(jié)果表明，50mol L–1的化合物1具有一定的乙酰膽堿酯酶抑制活性，其抑制率為(49.9±1.4)%，化合物2~7的抑制率均小于10.0%。

3 討論

本研究組前期報(bào)道了國產(chǎn)‘綠奇楠’沉香的化學(xué)成分主要是倍半萜和2-(2-苯乙基)色酮類化合物[6,10?13]，本研究從國產(chǎn)‘綠奇楠’沉香的乙醚和乙醇提取物中共分離鑒定出7個(gè)化合物，其中有1個(gè)倍半萜類化合物順式-7-羥基菖蒲烯 (1), 1個(gè)2-(2-苯乙基)色酮聚合物(5,6,7,8)-2-(苯乙基)-6,7,8-三羥基-5,6,7,8-四氫-5-[2-(2-苯乙基)色酮基-6-氧代]色酮 (2), 1個(gè)酚類化合物1-羥基-1,5-二苯基戊-3-酮(3), 1個(gè)木脂素苷丁香樹脂酚葡萄糖苷 (4), 1個(gè)三萜類化合物(3)-齊墩果-12-烯-3,23-二醇(5), 1個(gè)甾體類化合物-谷甾醇 (6)和1個(gè)脂肪酸類化合物棕櫚酸--單甘油酯 (7)，其中，化合物1、3~5和7均為首次從沉香中分離得到。

本研究組前期報(bào)道‘綠奇楠’沉香中具有乙酰膽堿酯酶抑制活性的艾里莫酚烷型倍半萜，而本研究分離出1個(gè)杜松烷型倍半萜：順式-7-羥基菖蒲烯(1)，也顯示出較好的抑制活性，且表現(xiàn)出非常甜的芳香氣味。值得一提的是，從沉香分離鑒定出的300余個(gè)化學(xué)成分中，僅有3個(gè)杜松烷型倍半萜[7]，且均不是從國產(chǎn)沉香中得到的，而倍半萜類化合物是沉香的兩大類主要成分之一，基于此，沉香中杜松烷型倍半萜還是有較大分離潛力的。在本研究中,我們還得到了(5,6,7,8)-2-(苯乙基)-6,7,8-三羥基-5,6,7,8-四氫-5-[2-(2-苯乙基)色酮基-6-氧代]色酮(2)，這是目前從‘奇楠’沉香中分離鑒定出的唯一1個(gè)2-(2-苯乙基)色酮聚合物[3?4,7,23?26]，而目前報(bào)道沉香中的2-(2-苯乙基)色酮聚合物約有40個(gè)[16,27?33],由此推斷，從‘奇楠’沉香繼續(xù)分離出2-(2-苯乙基)色酮聚合物的可能性是較大的。1-羥基-1,5-二苯基戊-3-酮 (3)在結(jié)構(gòu)上與沉香特征性沉香分2-(2-苯乙基)色酮類具有相關(guān)性，且據(jù)報(bào)道，此類化合物主要分布在瑞香科植物中[34]，本次也是首次從沉香樹脂中得到此類成分。此外，木脂素類化合物在未結(jié)香的白木部分大量分布[35?36]，而本研究首次在結(jié)香后的沉香樹脂中發(fā)現(xiàn)了1個(gè)木脂素苷丁香樹脂酚葡萄糖苷 (4)；同時(shí)，三萜和甾體類化合物在白木香的樹干、葉子、果實(shí)等部位均有分布，且-谷甾醇(6)也從白木香分離得到[35?36]。本研究分離得到的化合物3~7并不屬于沉香的兩類特征性成分倍半萜和2-(2-苯乙基)色酮，同時(shí)結(jié)合前人對(duì)未結(jié)香的白木部分的研究，不難推斷，這些成分很可能是殘存的白木部分的成分，也即雖然‘奇楠’沉香是品質(zhì)非常高的沉香，但其還是殘存白木部分的化學(xué)成分，這為后續(xù)高品質(zhì)‘奇楠’沉香的深入研究提供了一些參考。綜上，本研究果進(jìn)一步揭示了國產(chǎn)‘綠奇楠’沉香的化學(xué)成分和生物活性，為高品質(zhì)‘奇楠’沉香的后續(xù)研究和開發(fā)利用提供了一定的研究基礎(chǔ)。

[1] BARDEN A, AWANG ANAK N, MULLIKEN T A, et al. Heart of the Matter: Agarwood Use and Trade and CITES Implementation for[M]. Cambridge: TRAFFIC International, 2000: 1–51.

[2] YAMAGATA E, YONEDA K. Pharmacognostical studies on the crude drug of agarwood (VI): On “Kanankoh” [J]. Nat Med, 1987, 41(2): 142–146.

[3] NAKANISHI T, INADA A, NISHI M, et al. A new and a known derivatives of 2-(2-phenylethyl) chromone from a kind of agarwood (“Kanankoh,” in Japanese) originating from[J]. J Nat Prod, 1986, 49(6): 1106–1108. doi: 10.1021/np50048a024.

[4] ISHIHARA M, TSUNEYA T, UNEYAMA K. Fragrant sesquiterpenes from agarwood [J]. Phytochemistry, 1993, 33(5): 1147–1155. doi: 10. 1016/0031-9422(93)85039-T.

[5] XIE Z W. Discussion on Varieties of Chinese Medicinal Herb, Vol. 2 [M]. Shanghai: Shanghai Science and Technology Press, 1984.謝宗萬. 中藥材品種論述, 中冊[M]. 上海: 上?？茖W(xué)技術(shù)出版社, 1984.

[6] YANG D L, MEI W L, ZENG Y B, et al. 2-(2-Phenylethyl)chromone derivatives in Chinese agarwood “Qi-Nan” from[J]. Planta Med, 2013, 79(14): 1329–1334. doi: 10.1055/s-0033-1350647.

[7] DAI H F. Research Progress of Agarwood [M]. Beijing: Science Press, 2017. 戴好富. 沉香的現(xiàn)代研究[M]. 北京: 科學(xué)出版社, 2017.

[8] YANG D L, MEI W L, YANG J L, et al. GC-MS analysis of the fragrant sesquiterpenes and 2-(2-phenylethyl) chromone derivatives in four types of agarwood “Qi-Nan” [J]. Chin J Trop Crops, 2014, 35(6): 1235–1243. doi: 10.3969/j.issn.1000-2561.2014.06.033. 楊德蘭, 梅文莉, 楊錦玲, 等. GC-MS分析4種奇楠沉香中致香的倍半萜和2-(2-苯乙基)色酮類成分 [J]. 熱帶作物學(xué)報(bào), 2014, 35(6): 1235–1243. doi: 10.3969/j.issn.1000-2561.2014.06.033.

[9] FU L G. Red Book of Chinese Plant: Rare and Endangered Plants [M]. Beijing: Science Press, 1992: 670. 傅立國. 中國植物紅皮書——稀有瀕危植物 [M]. 北京: 科學(xué)出版社, 1992: 670.

[10] YANG D L, WANG H, GUO Z K, et al. A new 2-(2-phenylethyl) chromone derivative in Chinese agarwood ‘Qi-Nan’ from[J]. J Asian Nat Prod Res, 2014, 16(7): 770–776. doi: 10.1080/ 10286020.2014.896342.

[11] YANG D L, WANG H, GUO Z K, et al. Fragrant agarofuran and eremophilane sesquiterpenes in agarwood ‘Qi-Nan’from[J]. Phytochem Lett, 2014, 8: 121–125. doi: 10.1016/j.phytol. 2014.03.003.

[12] YANG D L, LI W, DONG W H, et al. Five new 5, 11-epoxyguaiane sesquiterpenes in agarwood “Qi-Nan” from[J]. Fitoterapia, 2016, 112: 191–196. doi: 10.1016/j.fitote.2016.05.014.

[13] SHAO H, MEI W L, KONG F D, et al. A new 2-(2-phenylethyl) chromone glycoside in Chinese agarwood “Qi-Nan” from[J]. J Asian Nat Prod Res, 2017, 19(1): 42?46. doi: 10.1080/ 10286020.2016.1187602.

[14] ELLMAN G L, COURTNEY K D, ANDRES V Jr, et al. A new and rapid colorimetric determination of acetylcholinesterase activity [J]. Biochem Pharmacol, 1961, 7(2): 91–95. doi: 10.1016/0006-2952(61) 90145-9.

[15] DAVILA-HUERTA G, HAMADA H, DAVIS G D, et al. Cadinane-type sesquiterpenes induced inby bacterial inocu- lation [J]. Phytochemistry, 1995, 39(3): 531–536. doi: 10.1016/0031- 9422(94)00958-V.

[16] IWAGOE K, KAKAE T, KONISHI T, et al. Studies on the agalwood (Jinko): VIII. Structures of bi-phenylethylchromone derivatives [J]. Chem Pharm Bull, 1989, 37(1): 124–128.

[17] NIWA M, JIANG P F, HIRATA Y. Constituents of: II. Absolute configurations of 1,5-diphenylpentane-1,3-diols [J]. Chem Pharm Bull, 1987, 35(1): 108–111.

[18] KOBAYASHI H, KARASAWA H, MIYASE T, et al. Studies on the constituents of: V. Isolation and structures of two new phenylpropanoid glycosides, cistanosides E and F [J]. Chem Pharm Bull, 1985, 32: 1452–1457.

[19] SONG Z H, MO S H, CHEN Y, et al. Studies on chemical constituents of(Schenk) R. Wight [J]. China J Chin Mat Med, 2000, 25(12): 728–730. 宋志宏, 莫少紅, 陳燕, 等. 管花肉蓯蓉化學(xué)成分的研究[J]. 中國中藥雜志, 2000, 25(12): 728–730.

[20] WANG K W, SUN H X, WU B, et al. Two novel olean triterpenoids from[J]. Helv Chim Acta, 2005, 88(5): 990–995. doi: 10.1002/hlca.200590094.

[21] CUI Y L, MU Q, HU C Q. Studies on the chemical constituents of[J]. Chin Pharm J, 2004, 39(3): 173–175. doi: 10.3321/ j.issn:1001-2494.2004.03.005. 崔益冷, 穆青, 胡昌奇. 紅花錦雞兒化學(xué)成分的研究[J]. 中國藥學(xué)雜志, 2004, 39(3): 173–175. doi: 10.3321/j.issn:1001-2494.2004.03.005.

[22] LU L M, SONG S J, WU Y, et al. Study on the chemical constituents of the ovum oil ofDavid (II) [J]. J Shenyang Pharm Univ, 2002, 19(1): 25–26. doi: 10.3969/j.issn.1006- 2858.2002.01.007. 盧立明, 宋少江, 武勇, 等. 哈士蟆卵油化學(xué)成分研究(II) [J]. 沈陽藥科大學(xué)學(xué)報(bào), 2002, 19(1): 25–26. doi: 10.3969/j.issn.1006-2858. 2002.01.007.

[23] ISHIHARA M, TSUNEYA T, SHIGA M, et al. Three sesquiterpenes from agarwood [J]. Phytochemistry, 1991, 30(2): 563–566. doi: 10. 1016/0031-9422(91)83727-3.

[24] ISHIHARA M, TSUNEYA T, UNEYAMA K. Guaiane sesquiterpenes from agarwood [J]. Phytochemistry, 1991, 30(10): 3343–3347. doi: 10. 1016/0031-9422(91)83206-Z.

[25] HASHIMOTO K, NAKAHARA S, INOUE T, et al. A new chromone from agarwood and pyrolysis products of chromone derivatives [J]. Chem Pharm Bull, 1985, 33(11): 5088?5091.

[26] UEDA J Y, IMAMURA L, TEZUKA Y, et al. New sesquiterpene from Vietnamese agarwood and its induction effect on brain-derived neurotrophic factor mRNA expression[J]. Bioorg Med Chem, 2006, 14(10): 3571?3574. doi: 10.1016/j.bmc.2006.01.023.

[27] YANG Y, CHEN H Q, KONG F D, et al. Dimeric sesquiterpenoid--chromone derivatives from agarwood ofand their cytotoxicity [J]. Phytochemistry, 2018, 145: 207?213. doi: 10. 1016/j.phytochem.2017.08.007.

[28] YANG Y, MEI W L, KONG F D, et al. Four new bi-2-(2-phenylethyl) chromone derivatives of agarwood from[J]. Fito- terapia, 2017, 119: 20?25. doi: 10.1016/j.fitote.2017.03.008.

[29] XIANG P, MEI W L, CHEN H Q, et al. Four new biphenylethyl- chromones from artificial agarwood [J]. Fitoterapia, 2017, 120: 61?66. doi: 10.1016/j.fitote.2017.05.012.

[30] HUO H X, ZHU Z X, SONG Y L, et al. Anti-inflammatory dimeric 2-(2-phenylethyl)chromones from the resinous wood of[J]. J Nat Prod, 2018, 81(3): 543?553. doi: 10.1021/acs.jnat prod.7b00919.

[31] IWAGOE K, KONISHI T, KIYOSAWA S, et al. The structures of AH10 and AH11, nobel bi-phenylethylchromones from agalwood [J]. Chem Pharm Bull, 1986, 34(11): 4889?4891.

[32] IWAGOE K, KODAMA S, KONISHI T, et al. The structures of AH15 and AH18, new bi- and tri-phenylethylchromones from agalwood [J]. Chem Pharm Bull, 1987, 35(11): 4680?4682.

[33] KONISHI T, IWAGOE K, KIYOSAWA S, et al. Tri-2-(2-phenylethyl) chromones from agalwood [J]. Phytochemistry, 1989, 28(12): 3548– 3550. doi: 10.1016/0031-9422(89)80391-7.

[34] PENG Y J, HUANG C S, LIU H X, et al. Recent advances in the structure and biological activity of 1,5-dihpenylpentadions [J]. Nat Prod Res Dev, 2009, 21(6): 1069–1075,1079. doi: 10.3969/j.issn.1001-6880.2009.06.041. 彭艷君, 黃初升, 劉紅星, 等. 1,5-二芳基戊烷類化合物生物活性與結(jié)構(gòu)關(guān)系的研究進(jìn)展 [J]. 天然產(chǎn)物研究與開發(fā), 2009, 21(6): 1069– 1075,1079. doi: 10.3969/j.issn.1001-6880.2009.06.041.

[35] LI W, CAI C H, DONG W H, et al. 2-(2-Phenylethyl)chromone derivatives from Chinese agarwood induced by artificial holing [J]. Fitoterapia, 2014, 98: 117–123. doi: 10.1016/j.fitote.2014.07.011.

[36] LI W, MEI W L, ZUO W J, et al. Advances in chemical constituents and biological activities from[J]. J Trop Subtrop Bot, 2014, 22(2): 201–212. doi: 10.3969/j.issn.1005-3395.2014.02.015. 李薇, 梅文莉, 左文健, 等. 白木香的化學(xué)成分與生物活性研究進(jìn)展[J]. 熱帶亞熱帶植物學(xué)報(bào), 2014, 22(2): 201–212. doi: 10.3969/ j.issn.1005-3395.2014.02.015.

Study on Chemical Constituents of Chinese Agarwood ‘Qi-Nan’

LI Wei, MEI Wen-li, DONG Wen-hua, CAI Cai-hong, GAI Cui-juan, DAI Hao-fu*

(Key Laboratory of Biology and Genetic Resources of Tropical Crops, Ministry of Agriculture, Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Hainan engineering research center of agarwood,Haikou Key Laboratory for Research and Development of Tropical Natural Products, Haikou 571101, China)

To understand the chemical constituents of Chinese agarwood “Qi-Nan”, seven compounds were isolated by various column chromatographic methods. On the basis of spectral data, they were identified as-7- hydroxycalamenene (1), (5,6,7,8)-2-(phenylethyl)-6,7,8-trihydroxy-5,6,7,8-tetrahydro-5-[2-(2-pheny-lethyl) chromonyl-6-oxy]chromone (2), 1-hydroxy-1,5-diphenylpentan-3-one (3),syringaresinol-glycoside(4), (3)- olean-12-ene-3,23-diol(5),-sitosterol (6) and hexadecanoic acid, 2,3-dihydroxypropyl ester (7). Compounds 1, 3-5, and 7 were isolated from agarwood for the first time, and compound 1 showed a very sweet aromatic odor. Compound 1 showedacetylcholinesterase (AChE) inhibitory activity with the inhibition rate of (49.9±1.4)% at the concentration of 50mol L–1.

‘Qi-Nan’; Chinese agarwood; Chemical constituent; Acetylcholinesterase (AChE); Inhibitory activity

10.11926/jtsb.3958

2018-06-11

2018-07-13

海南自然科學(xué)基金創(chuàng)新研究團(tuán)隊(duì)項(xiàng)目(2017CXTD020)；中國熱帶農(nóng)業(yè)科學(xué)院院級(jí)創(chuàng)新團(tuán)隊(duì)南藥產(chǎn)業(yè)技術(shù)創(chuàng)新團(tuán)隊(duì)(17CXTD-15)；現(xiàn)代農(nóng)業(yè)產(chǎn)業(yè)技術(shù)體系建設(shè)專項(xiàng)(CARS-21)資助

This work was supported by Innovative Research Team Grant of the Natural Science Foundation of Hainan Province (Grant No. 2017CXTD020), Central Public-interest Scientific Institution Basal Research Fund for Innovative Research Team Program of CATAS (Grant No. 17CXTD-15), and the Special Project for China Agriculture Research System (Grant No. CARS-21).

李薇(1988~)，女，助理研究員，從事天然產(chǎn)物化學(xué)研究。E-mail: liwei@itbb.org.cn

E-mail: daihaofu@itbb.org.cn