霍少杰 吳繼春 杜江
【摘 要】 目的:研究苗藥三兩銀(Sarcococcahookeriana)的生物堿類成分。方法:采用溶劑提取、各種色譜分離技術(shù)進(jìn)行化合物純化,通過波譜數(shù)據(jù)對(duì)單體化合物進(jìn)行結(jié)構(gòu)鑒定。結(jié)果:分離鑒定了三個(gè)化合物,均為孕甾烷生物堿,分別為:Pachyaximine (A) (1)、(20S)-20-(N,N-dimethylamino)-16α,17α-epoxy-3β- methoxy-pregn-5-ene (2)、(2α,3β,4β,20S)-2,4-bis(acetoxy)-20-(N,N-dimethylamino)- 3-[(3-methylbut-2-enoyl)amino]-5α-pregnane (3)。結(jié)論:三個(gè)化合物均為首次從苗藥三兩銀中分離得到。
【關(guān)鍵詞】 苗藥;三兩銀;化學(xué)成分;孕甾烷生物堿
【中圖分類號(hào)】R284.1 【文獻(xiàn)標(biāo)志碼】 A【文章編號(hào)】1007-8517(2018)02-0039-03
Abstract:Objective To study the chemical constituents from Miao medicine SarcococcahookerianaLindl. Method The chemical constituents from S. hookeriana were isolated by solvent extraction and chromatographic separation technology. Structures of obtained compounds were elucidated by modern spectroscopy. Results Three known steroidal alkaloids were isolated and identified as Pachyaximine (A) (1), (20S)-20-(N,N-dimethylamino)-16α,17α-epoxy-3β-methoxy-pregn-5-ene (2), and (2α,3β,4β,20S)-2,4-bis(acetoxy)-20-(N,N-dimethylamino)-3-[(3-methylbut-2-enoyl)amino]-5α-pregnane (3). Conclusion All the compounds were isolated from this plant for the first time.
Keywords:Miao medicine; Sarcococcahookeriana; Chemicalconstitutents; Pregnane-type Steroidal Alkaloids
羽脈野扇花(Lindl Sarcococcahookeriana.)為黃楊科(Buxaceae)野扇花屬植物,常綠灌木。本屬約有20種,分布于亞洲東部和南部,我國(guó)已知有8種及幾個(gè)變種[1],是傳統(tǒng)的民族民間用藥。羽脈野扇花和野扇花(S. ruscifolia Stap)同為貴州苗族民間習(xí)用藥材,?;煊茫置齼摄y、三兩金、羊不食等,用于治療胃潰瘍、胃痛、腹瀉、發(fā)熱、咽喉痛、跌打損傷、腦憨癥等疾病[2-3]。野扇花屬植物的化學(xué)成分研究始于上世紀(jì)60年代,從中發(fā)現(xiàn)大量孕甾烷生物堿,以及三萜、甾體、糖苷等類型化合物。迄今已從該屬植物中分離得到100余個(gè)孕甾烷生物堿[4],它們中的大多數(shù)化合物因具有明顯的膽堿酯酶抑制活性而受到各國(guó)學(xué)者的廣泛關(guān)注[5-9],有望從中發(fā)現(xiàn)新的治療阿爾茨海默病的先導(dǎo)化合物。從羽脈野扇花中還分離到一系列具有抗利什曼蟲[10-12]、抗瘧原蟲[9]、抗菌[11-12]、細(xì)胞毒[13-14]、抗炎[15]、免疫抑制[16]及促雌激素生成[17]等活性的成分。本課題組前期對(duì)野扇花和羽脈野扇花中的化學(xué)成分進(jìn)行了初步研究,并對(duì)研究成果做了相關(guān)的文獻(xiàn)報(bào)道[18,19]。為了深入研究該屬植物的活性成分,課題組繼續(xù)對(duì)黔產(chǎn)羽脈野扇花的生物堿成分進(jìn)行了分離研究。
1 儀器與材料
1.1 儀器 JEOL ECX-400導(dǎo)核磁共振譜儀(日本電子株式會(huì)社);R-215型旋轉(zhuǎn)蒸發(fā)儀(瑞士步琦);ZF-2型紫外光譜儀(上海安亭電子儀器廠);RE-5210旋轉(zhuǎn)蒸發(fā)器(上海亞榮生化儀器廠);SHZ-DⅢ型多用真空泵(鄭州華儀器有限責(zé)任公司);SK8210HP超聲儀(上??茖?dǎo)超聲儀器有限公司)。
1.2 材料 層析硅膠:(200-300目、300-400目)青島海洋化工有限公司;顯色劑:改良碘化鉍甲試劑;其它試劑均為國(guó)產(chǎn)分析純。
實(shí)驗(yàn)藥材于2015年7月采于貴州省赫章縣韭菜坪,由貴陽中醫(yī)學(xué)院趙能武教授鑒定為羽脈野扇花(Baill Sarcococcahookeriana.)。
2 提取分離
羽脈野扇花根莖(14.5 kg)自然干燥后粉碎,用95%甲醇超聲提取四次,提取液減壓回收溶劑,濃縮后得總提取物??偺崛∥镉?% H2SO4溶液調(diào)pH到1,乙酸乙酯萃取除去脂溶性雜質(zhì),剩余酸水液用飽和Na2CO3溶液調(diào)pH至9,二氯甲烷萃取得總生物堿156 g。總生物堿進(jìn)行硅膠柱層析,二氯甲烷-甲醇(100∶0→5∶1)梯度洗脫,薄層層析檢測(cè),分為A-E五個(gè)部分。A部分進(jìn)行硅膠柱層析,石油醚-二氯甲烷-二乙胺(100∶1∶1→5∶1∶1)梯度洗脫,分為A1-A5五個(gè)部分。A1部分通過反復(fù)硅膠柱層析(分別以石油醚-二氯甲烷-二乙胺和二氯甲烷-甲醇為洗脫系統(tǒng))得化合物1(188 mg)和2(76 mg)。B部分進(jìn)行硅膠柱層析,二氯甲烷-甲醇(100∶1→10∶1)梯度洗脫,分為B1-B3三個(gè)部分。B3部分通過反復(fù)硅膠柱層析(分別以石油醚-二氯甲烷-二乙胺和二氯甲烷-甲醇為洗脫系統(tǒng))得化合物3(46 mg)。
3 結(jié)構(gòu)鑒定
化合物1白色細(xì)針狀晶體,易溶于氯仿,改良碘化鉍鉀顯色劑顯色呈橘紅色。1H-NMR (CDCl3, 400MHz) δ: 5.36 (m, 1H, H-6), 3.36 (s, 3H, OCH3), 3.07 (tt, J = 11.3, 4.5 Hz, 1H, H-3), 2.17 (s, 3H, NMe2), 1.00 (s, 3H, H-19), 0.88 (d, J = 6.4 Hz, 3H, H-21), 0.68 (s, 3H, H-18). 13C NMR (CDCl3, 100MHz) δ: 140.8 (C-5), 121.6 (C-6), 80.3 (C-3), 61.1 (C-20), 56.9 (C-14), 55.6 (OMe), 54.7 (C-17), 50.1 (C-9), 41.4 (C-13), 39.9 (NMe2), 39.5 (C-12), 38.6 (C-4), 37.2 (C-1), 36.9 (C-10), 31.9 (C-8), 31.8 (C-7), 28.0 (C-2), 27.7 (C-16), 24.1 (C-15), 21.1 (C-11), 19.4 (C-19), 12.1 (C-18), 9.8 (C-21)。以上數(shù)據(jù)與文獻(xiàn)[20]報(bào)道的NMR數(shù)據(jù)基本一致,故鑒定化合物1為Pachyaximine (A)。
化合物2 無色細(xì)針狀晶體,易溶于氯仿,改良碘化鉍鉀顯色劑顯色呈橘紅色。H-NMR (CDCl3, 400MHz) δ:5.35 (m, 1H, H-6), 3.61 (s, 1H,H-16), 3.35 (s, 3H,OMe),3.06 (tt, J = 11.3, 4.5 Hz, 1H,H-3),2.89 (d, J= 6.4 Hz, 1H,H-20), 2.28 (s, 6H,NMe2),1.01 (s, 3H,H-19), 0.91 (s, 3H,H-18),1.17 (d,J =6.7 Hz, 3H, H-21). 13C-NMR (CDCl3, 100MHz) δ:141.1(C-5), 121.4(C-6), 80.3(C-3), 73.6(C-17), 61.1(C-16), 56.0(C-20), 55.8(OMe), 50.5(C-9), 45.6(C-14), 43.2(NMe2), 42.1(C-13), 38.8(C-4), 37.2(C-10), 37.2(C-1), 32.9(C-12), 31.7(C-7), 30.4(C-8), 28.1(C-2), 27.4(C-15), 20.8(C-11), 19.5(C-19), 15.8(C-18), 13.6(C-21)。以上數(shù)據(jù)與文獻(xiàn)[17]報(bào)道的NMR數(shù)據(jù)基本一致,故鑒定化合物2為(20S)-20-(N,N-dimethylamino)-16α,17α-epoxy-3β-methoxy-pregn-5-ene。
化合物3白色細(xì)針狀晶體易溶于氯仿,改良碘化鉍鉀顯色劑顯色呈橘紅色。H-NMR(CDCl3,400MHz)δ: 5.62 (d,J=8.9 Hz, 1H,NH),5.50(s,1H,H-2′),5.26(t,J=2.8Hz,1H,H-4),5.11(d,J=3.6Hz,1H,H-2),4.32(dt,J=8.4,4.0Hz,1H,H-3),2.46(t,J=8.6Hz,1H),2.10(s,3H,MeCO),2.08(s,3H,MeCO),1.12(s,3H,H-19),0.89(d,J=6.5Hz,3H,H-21),0.63(s,3H,H-18). 13C-NMR (CDCl3,100MHz)δ:170.7(MeCO), 170.5(MeCO), 166.1(C-1′), 152.2(C-3′), 118.1(C-2′), 74.2(C-4), 72.0(C-2), 61.4(C-20), 56.4 (C-9), 55.7(C-17), 49.3(C-3), 48.8(C-5), 41.8(C-13), 40.9(C-1), 39.9(NMe2), 39.6(C-12), 34.9(C-8), 34.8(C-10), 31.8(C-7), 27.7(C-16), 27.3(C-6), 25.2(C-15), 24.1(C-5′), 21.5(C-11), 21.2(MeCO), 20.7(MeCO), 20.0(C-4′), 15.4(C-19), 12.4(C-18),10.1(C-21)。以上數(shù)據(jù)與文獻(xiàn)[21]報(bào)道的NMR數(shù)據(jù)基本一致,故鑒定化合物3為(2α,3β,4β,20S)-2,4-bis(acetoxy)-20-(N,N-dimethylamino)-3-[(3-methylbut-2-enoyl)amino]-5α-pregnane。
4 討論
野扇花屬植物在貴州省資源豐富,民間使用廣泛,國(guó)外學(xué)者從該屬植物中分離出大量的孕甾烷生物堿,然而關(guān)于該屬植物的化學(xué)成分研究,國(guó)內(nèi)報(bào)道較少。本實(shí)驗(yàn)從黔產(chǎn)苗藥三兩銀中分離鑒定了3個(gè)孕甾烷生物堿,分別為Pachyaximine (A) (1)、(20S)-20-(N,N-dimethylamino)-16α,17α-epoxy-3β- methoxy-pregn-5-ene (2)、(2α,3β,4β,20S)-2,4-bis(acetoxy)-20-(N,N-dimethylamino)-3-[(3-methylbut-2-enoyl)amino]-5α-pregnane (3),化合物結(jié)構(gòu)見圖1。3個(gè)化合物均為首次從該植物中分離得到。
苗族人民在長(zhǎng)期與疾病斗爭(zhēng)的過程中形成了獨(dú)特的診療體系,在治療外傷和一些地方性多發(fā)病方面往往有獨(dú)到的效果。課題組在民間調(diào)研時(shí)發(fā)現(xiàn)野扇花屬植物在苗族民間廣泛應(yīng)用于治療胃腸潰瘍、腦憨癥等疾病。結(jié)合三兩銀在民間醫(yī)藥中的應(yīng)用,本實(shí)驗(yàn)對(duì)其進(jìn)行系統(tǒng)的化學(xué)成分和生物活性研究,為合理開發(fā)苗藥三兩銀的藥用價(jià)值提供理論基礎(chǔ)。
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