全 文 ：Water-solub le phenolic glycosides from the whole plant
ofBulbophyllum odoratissimum
LIU Dai-lin1, 3 , PANG Fa-gen1 , ZHANG Xue1 , GAO H ao1 , WANG N ai-li1, 2 , YAO X in-sheng1, 2*
(1. Departm ent ofNatura l ProductsChem istry, Shenyang Pharm aceutica lUniversity, Shenyang 110016 , China;
2. Shenzhen Research Center of T rad itiona l ChineseMedicine and Natural P roducts, Shenzhen 518057 , China;
3. Departm ent of Pharm acognosy, M ed ical College of ChineseP eople’ s Armed PoliceF orces, Tian jin 300162 , Ch ina)
Abstract:A mi 　 To investiga te the w ater-soluble pheno lic g ly cosides from the w ho le p lant o f
Bu lbophy llum odoratissimum. Methods　Co lumn ch roma tography techniques w ere used to iso la te the
chem ica l constituents, phy sico-chem ical constants and spectroscop ic ana lysis w ere emp loyed for structu ra l
e lucidation. Results　Bu lbophy llinoside (1), a new pheno lic g lycoside and th ree known compoundsw ere
iso lated from the who le plan t ofBu lbophy llum odoratissimum Lind.l Their struc turesw e re de term ined as 3-
hydroxyphenethy l a lcohol 4-O-(6′-O-β-apio fu ranosy l)-β-D-g lucopy ranoside (1), 3-me thoxyphenethy l
a lcoho l 4-O-β-D-g lucopynanoside (2), 3, 5-dimethoxyphenethy l a lcohol 4-O-β-D-g lucopynanoside (3)
and sy ring in (4). Conc lusion　Bu lbophy llinoside (1) is a new compound.
Key words:Bu lbophy llum odoratissimum;pheno lic g lycosides;bu lbophy llino side
CLC num ber:R284. 1;R284. 2　　　Docum ent code:A　　　A rticle ID:0513 -4870(2006)08 - 0738 -04
Received date:2005-12-29.
* Correspond ing au thor　Tel /Fax:86 - 20 - 85225849,
E-m ail:yaoxinsh eng@ vip. tom. com
密花石豆兰中水溶性酚酸类成分研究
刘岱琳 1, 3 , 庞发根 1 , 张　雪 1 , 高　昊 1 , 王乃利 1, 2 , 姚新生 1, 2*
(1. 沈阳药科大学 天然药物化学教研室 , 辽宁 沈阳 110016;2. 深圳中药及天然药物研究中心 , 广东 深圳 518057;
3. 天津武警医学院 生药教研室 , 天津 300162)
摘要:目的　对密花石豆兰中的水溶性酚酸类成分进行研究。方法　采用大孔树脂 HP20, Sephadex LH-20和
ODS等多种柱色谱方法进行分离 , 利用化合物的理化常数并结合波谱解析鉴定化合物的结构。结果　从密花石豆
兰的水溶性成分中共分离得到 4个化合物 , 分别鉴定为 3-羟基-苯乙醇-4-O-(6′-O-β-芹呋喃糖基 )-β-D-葡吡喃糖苷
(1), 3-甲氧基-苯乙醇-4-O-β-D-葡吡喃糖苷 (2), 3, 5-二甲氧基苯乙醇-4-O-β-D-葡吡喃糖苷(3), sy ringin (4)。结论
化合物 1为新化合物 , 命名为 bu lbophy llinoside。
关键词:密花石豆兰;酚苷;bu lbophy llinoside
　　Prev ious chem ical studies show ed bibenzy ls[ 1 -4] ,
phenanth renes
[ 5 -7]
and 9, 10-dihydrophenan threnes[ 8, 9]
and the ir dimmers
[ 10, 11]
we re thema in components from
theO rch idaceae. Bu t the po lar constituents, especially
the w ater-so luble constituents, have rare ly been
exam ined. Con tinuing our w ork on the constituents of
Bu lbophy llum odoratissimum Lind.l , we have focused
on the w ater-so luble constituents of the p lant, and have
iso lated a new pheno lic g lyco side named bulbophy llino-
side, as w e ll as three known compounds. W e report
he re the iso lation and structural elucidation o f these
pheno lic glycosides obta ined from the whole plant o f
B. odoratissimum Lind.l
The n-bu tano l so luble fraction o f the e thano l
extract o f Bulbophy llum odoratissimum was applied on
D iaionHP20 co lumn ch roma tography e lu ted w ithH2O-
E tOH gradiently. Fou r w a ter-so lub le glycosides w ere
iso lated and purified by a comb ination o f chromato-
g raphic methods from the 30% E tOH so luble fraction.
Compounds 2 - 4were identified as 3-methoxyphenethy l
738 药学学报 Ac ta Pharm aceu tica S in ica 2006, 41(8):738 - 741
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alcoho l 4-O-β-D-g lucopynanoside (2), 3, 5-dime tho-
xyphenethy l alcoho l 4-O-β-D-glucopynanoside (3) and
sy ring in (4), based on the ir spectral da ta and
comparison w ith literature da ta
[ 12] (Figu re 1).
Compound 1 was obtained as wh ite amo rphous
powderw hosemo lecular formula w as de te rm ined to be
C19H28O 12 from the obse rva tion of a quasi-[M +Na] +
by high reso lu tion E I-m ass spectrometry. On acid
hydro lysis of compound 1, apiose and g lucose w ere
identified by co-TLC w ith au thentic samples. The peak
atm /z 317[M +Na - sugar] + in the positive-ion ES I-
MS
2
and the peak atm /z 293 [M -H - sugar] - in the
nega tive-ion ES I-MS2 also revealed the presence o f an
apiosemo iety. The IR spectrum indicated the presence
of hydroxy l g roups (3 300 cm -1), an a roma tic ring
(1 595 and 1 510 cm -1) and pheno lic hydroxyl g roups
(1 210 cm - 1);the UV absorption m aximas ind ica ted
the presence of an aroma tic moie ty. The
1
H NMR
spectrum in CD3OD show ed the presence of three
aromatic protons [ δ7.09(1H , d, J =8.2 H z), 6.73
(1H , d, J =2.1 H z) and 6.65(1H , dd, J =8.2, 2.1
H z)] coupled in an ABX-system and tw o anomeric
pro tons from sugars a tδ4.99(1H , d, J =2.4 Hz) and
4.65(1H , d, J =7.5 H z). O the r signals o f four
pro tons a tδ3.69(2H , t, J =7.1Hz) and 2.70(2H , t,
J =7.1H z) in the up fie ld region w e re assigned to the
fragment-CH2CH2-by 1H-1H COSY. Fu rther info rma-
tion ob tained from the
13
C NMR and HMBC spectra
indica ted that the ag lycone portion must be 3, 4-
dihydroxy-phene thy l alcoho .l The sequence of the
oligo saccharide chain w as deduced by comparing the
chem ical shifts o f their indiv idua l sugar residues w ith
published compounds
[ 13] , and confirmed by HMBC
and NOESY experiments. The C-1 of g lucose w as
attached to C-4 o f the ag ly cone, as indicated by the
co rrela tion betw eenH-1 (δ4.65) o f glucose w ith C-4
(δ145.2) of the ag lycone inHMBC spectrum. In the
same w ay, the corre lation between H-1 (δ4.99) of
apiose w ith C-6 (δ68.2) of g lucose in the HMBC
spectrum suggested that apiosemoie ty must be attached
to C-6 of glucose. From the re lative ly large H-1
coupling constants(7.5H z), the anome ric hydroxy l of
glucose moie ty should be β-configuration. Based on the
comparison of the signa ls from apiose of compound 1
w ith that in the literature
[ 14] , the anomeric
configuration of apiose w as dete rm ined to be a β-
apio fu ranose. O n the basis o f the above evidence, the
struc ture o f compound 1 was elucidated as 3-
hydroxyphenethy l alcoho l 4-O-(6′-O-β-apiofuranosyl)-
β-D-glucopy ranoside, named as bulbophy llinoside. A s
shown in Table 1, comple te assignment o f 1 was
achieved w ith the aid of the
1
H-1H COSY , HMQC and
HMBC spectra.
Expermi ental section
ESI-MSwere conduced using B ruker esquire 2000
mass spectrome ter. IR spectra w ere reco rded on
Shimadzu FT /IR-8400 spectrome ter. 1H NMR and
13
C NMR , along w ith 2D NMR spectra w ere de ter-
m ined on a Bruke r AV-400 (400 MH z fo r 1H , 100
MHz for
13
C) NMR spectrome ter, using TM S as an
interna l standard. Chem ica l shifts we re expressed in δ
and coupling constants (J ) we re reported in Hertz
(Hz). TLC w as carried out on Silica gel 60F254 and
the spotsw ere v isualized by spray ing w ith 10%H 2SO 4
and hea ting. D ia ion HP-20 (M itsubishi Kasei),
Sephadex LH-20 (M itsub ish iK ase i) and ODS (40 -
63μm , Merck)were used for co lumn chroma tography.
Prepara tiveHPLC was perfo rmed using anODS co lumn
(C18 , 250 mm ×20 mm , Sh imadzu Pak;De tec to r:
UV)w ith SPD-10AVP UV-V IS de tec to r.
The who le plant of Bu lbophy llum odora tissimum
Lindlw e re collected from Guangxi prov ince o fCh ina in
2000, and identified by D r. L iping X iao. A voucher
specimen (YL-2001-75) is deposited in the Shenyang
PharmaceuticalUnive rsity, Liaoning Prov ince of China.
1　Extrac tion and iso lation
A ir-dried w ho le plan ts ofBulbophy llum odora tissi-
mum Lind l (3 kg) were ex tracted w ith 60% E tOH
(10 l×2) under reflux. The E tOH ex tract (280 g)
was then suspended in w a te r and subsequently
partitioned w ith cyc lohexane, ethy l ace tate and n-
bu tano l, respectively. The n-bu tano l extrac t (30 g)
was subjected to D iaionHP-20 chroma tography using a
g radient from H 2O to 95% E tOH. The fraction eluted
w ith 30% E tOH (7.4 g)was sepa ra ted by a Sephadex
LH-20 open co lumn w ith 40% MeOH to yie ld three
frac tions ( fraction 1-3). F raction 2 (4.2 g) was
further separated by an ODS open column w ith H2O /
M eOH gradient elution to y ield four sub fractions 21 -
24. Sub fraction 21 (640mg) e lu ted w ith 20%MeOH
was finally separated by PHPLC w ith M eOH-H 2O
(15∶85) to obta in 1 (10 mg) and 2 (12 mg).
Sub fraction 22 (716 mg) was finally separated by
PHPLC w ithM eOH-H 2O (1∶4) to afford 3 (20 mg)
and 4 (14 mg).
Bulbophyllinoside (1)　Amorphous powder. IR
υmax cm - 1:3 300, 2 900, 1 595, 1 510, 1 210, 1 165,
739 L IU Dai-lin, e t a l:W ater-so luble pheno lic g lycosides from the who le plan t o fB ulbophy llum odoratissim um
F igure 1　Structures of compounds 1 -4
Table 1　1H NMR and 13C NMR data for compounds 1 -4 in CD3OD (δ)
No.
Com pound 1
C arbon P roton
Compound 2
Carbon Proton
C om pound 3
C arb on P roton
Com pound 4
Carbon Proton
1 136. 2 135. 6 137.5 135. 3
2 117. 7 6. 73, d, (2.1) 114. 7 6.88, d, (1. 9) 108.0 6. 57, s 105. 5 6. 74, s
3 148. 3 150. 8 154.1 154. 4
4 145. 2 146. 5 134.8 135. 9
5 119. 3 7. 09, d, (8.2) 118. 4 7.07, d, (8. 2) 154.1 154. 4
6 121. 5 6. 65, dd, (8. 2, 2. 1) 122. 5 6.75, dd, (8. 2, 1. 9) 108.0 6. 57, s 105. 5 6. 74, s
7 39. 7 2. 07, 2H , t, (7. 1) 39. 8 2.76, 2H , t, (7. 0) 40.4 2. 75, 2H , t, (6. 8) 131. 3 6. 54, d, (15.8)
8 64. 3 3. 69, 2H , t, (7. 1) 64. 3 3.72, 2H , t, (7. 0) 64.1 3. 75, m;3. 73, d, (6.8) 130. 1 6. 30, d t, (15. 8, 5. 6)
9 63. 6 4. 21, dd, (5.6, 1. 4)
3-OCH
3 56. 7 3.84, 3H , s 57.0 3. 83, 6H , s 57. 0 3. 85, 3H , s
5-OCH3 57.0 3. 83, 6H , s 57. 0 3. 85, 3H , s
G lu-1　 104. 7 4. 65, d, (7.5) 103. 1 4.82, o 105.6 4. 85, d, (7. 4) 105. 4 4. 85, m
2 74. 9 3. 46, o 74. 9 3.45, o 75.7 3. 45, m 75. 7 3. 46, m
3 77. 6 3. 45, o 78. 2 3.37, o 78.3 3. 19, m 78. 4 3. 21, m
4 71. 6 3. 35, m 71. 4 3.37, o 71.3 3. 39, o 71. 4 3. 40, o
5 77. 2 3. 51, m 77. 9 3.45, o 77.8 3. 40, o 77. 8 3. 41, o
6 68. 2 4. 01, dd, (11. 1, 1. 9) 62. 5 3.86, o;3. 68, m 62.6 3. 78, o 62. 6 3. 77, dd, (11. 9, 2. 4)
3. 63, dd, (11. 1, 6. 5) 3. 65, dd, (12. 0, 5. 1) 3. 65, dd, (11. 9, 5. 1)
A pi-1　 111. 1 4. 99, d, (2.4)
2 78. 1 3. 92, d, (2.4)
3 80. 5
4 75. 0 3. 97, d, (9.7)
3. 75, d, (9.7)
5 65. 6 3. 58, 2H , s
a:A ll o f the signa ls we re a ssigned by 1H-1H COSY, HMQC and HMBC spectra. Overlapped signa ls are ind ica ted by “ o”;b:J
va lue s( in parenthe ses) are repo rted in H z
1 072, 1 010, 825. UV(MeOH) λmax nm( logε):206
(3.95), 218(3.75), 278(3.26). HRE I-MS(m /z):
[ M +Na ] + 471.150 4 ( ca lcd for C19H28O12Na,
471.147 8). Positive ES I-MS(m /z):471, 317, 229.
N egative ESI-MS(m /z):447 , 293, 233, 191, 161,
153, 123, 99. 1H NMR and 13C NMR data (Tab le 1).
3-M ethoxyphenethyl alcohol 4-O-β-D-gluco-
pynanoside (2)　White pow der. Positive ESI-MS
(m /z):353, 338, 307, 185. 1H NMR and 13C NMR
data (Tab le 1).
3, 5-Dmi ethoxyphene thyl a lcohol 4-O-β-D-
glucopynanoside (3)　Wh ite pow der. Po sitive ESI-
MS (m /z):383, 353, 221, 189, 185. 1H NMR and
13
C NMR data (Table 1).
Syr ing in (4)　W hite pow de r. Positive ESI-MS
(m /z):383. Nega tive ESI-MS(m /z):371. 1H NMR
740 药学学报 Ac ta Pharm aceu tica S in ica 2006, 41(8):738 - 741
and
13
C NMR data (Table 1).
2　Ac id hydrolysis
Bulbophy llinoside (2 mg) was heated w ith 6%
aq. HC l at 80 ℃ for 5 h. The reac tion m ix ture w as
neu tra lized w ith 1 mo l L - 1 aq. NaOH and filte red.
The filtrate w as partitioned w ith e thy l aceta te. The
aqueous layerw as concentrated and apiose and g lucose
w ere identified by TLC in compa rison w ith authentic
samp les using so lven t system pyridine-E tOAc-E tOH-
H2O (5∶5∶1∶3).
Acknow ledgem ents:The au thors g ra tefully acknow ledge
D r. L ip ing X iao of Kadoorie Fa rm ＆ Bo tanic Garden in
H ongkong for iden tification o f the p lan.t Thanks are a lso
ex tended to Shangha i Institute o fM ateriaMedica o f CAS fo r their
assistance in the HRES I-M S.
References
[ 1] M ajumder PL, Basak M. Two bibenzy l deriva tives from
the orch idCirrhopetalum anderson ii [ J] . Phy tochem istry,
1991, 30:321 -324.
[ 2] M ajumder PL, Ghosal S. Two stilbeno ids from the orch id
Arundina bam busifolia [ J] . Phytochem istry, 1993, 32:
439 - 444.
[ 3] M ajumder PL, Pal S. Cumu la tin and tristin, tw o b ibenzy l
deriva tive s from the o rch idsD endrobium cum ula tum and
Bulbophy llum triste [ J] . Phy tochem istry, 1993, 32:
1561 -1565.
[ 4] M ajumder PL, Ghosal S. Two stilbeno ids from the orch id
Arundina bam busifolia [ J] . Phytochem istry, 1994, 35:
205 - 208.
[ 5] M ajum de r PL, Basak M. C irrhope talin, a phenanthrene
de riva tive from Cirrhopetalum anderson ii [ J] . Phyto-
chem istry, 1990, 29:1002 -1004.
[ 6] M ajum de r PL, Sen RC. Pendu lin, a po lyoxygenated
phenanth rene derivative from the o rchid Cym bidium
pendulum [ J] . Phy tochem istry, 1991, 30:2432 - 2434.
[ 7] Yuan-W ah L, Leslie JH , Andrew DP. Phenanthrene and
o ther arom atic constituents of Bu lbophy llum vaginatum
[ J] . Phy tochem istry, 1999, 50:1237 -1241.
[ 8] M ajum de r PL, Lahiri S. Lusianthrin and lusianthrid in,
tw o stilbeno ids from the orch id Lusia indivisa [ J] .
Phy to chem istry, 1990, 29:621 -624.
[ 9] M ajum de r PL, Basak M. Two stilbenoids from the o rchid
Cirrhopeta lum andersonii [ J] . Phy tochem istry, 1991, 30:
3429 - 3432.
[ 10] M ajumde r PL, Pa l A, Joarda rM. C irrhopeta lanthrin, a
dim eric phenanth rene derivative from the o rchid
Cirrhopeta lum macu losum [ J] . Phy tochem istry, 1990,
29:271 - 274.
[ 11] M ajumde rPL, Lahiri S. Two stilbeno ids from the o rchid
E ria flava [ J] . Phy tochem istry, 1990, 29:3052 - 3055.
[ 12] M arina DG , M aria F, An tonio F, et a.l An tia lga l
com pounds from Zan tedeschia aethiopica [ J ] .
Phy to chem istry, 1998, 49:1299 - 1304.
[ 13] Otsuka H , Takeda Y, Yam asak i K. G lycosides of
m egastigmane and o f the sim ple a lcoho ls from Alangium
premn ifolium [ J] . Phy to chem istry, 1996, 42:723 -727.
[ 14] H idehiko K, Tosh inori I, H ideake O, e t a.l W a ter-
so luble pheno lic g lycosides from leaves of Alangium
premn ifolium [ J] . Phy tochem istry, 1997, 44:1551 -
1557.
基于实例的药物设计新平台
美国 CD I(Com puDrug Interna tiona l)开发的 CompuD rug药物设计软件包的引进 ,给国内药物设计 、药物合成专家提供了
研究新平台。目前 ,北京美康生物技术研究中心作为美国 CDI药物设计软件中国及香港地区推广的总代理 , 承担着业务培
训 、技术推广工作。
美国 CD I成立于 1983年 , 是最早建立药物研发软件平台的公司之一。 美国 CD I由 Com pudrug公司 (致力于药物
ADM E /Tox理化参数的预测), ComG enx公司(致力于药物设计及合成)和 C omGrid公司(致力于化学软件开发)三家国际公
司共同组成。从 1983年开始 ,美国 CDI便致力于与 ADME /Tox有关的理化参数的预测和研发。
Com puDrug软件包括:Pa llas系统和 EM IL工作组。它们是基于实例的药物研发的重要工具:Pa llas系统是关于
ADM E /Tox理化参数 、代谢产物和毒性预测的专业软件 , 帮助科研人员对所合成或设计的药物的理化参数 、代谢产物和毒
性进行预测 ,使该药物更加符合药物的 ADM E /Tox过程。 EM IL工作组是基于实例的关于药物先导化合物优化和演化的专
业软件 ,结合 Pa llas系统帮助科研人员设计具有良好 ADME /Tox特性的药物。
741 L IU Dai-lin, e t a l:W ater-so luble pheno lic g lycosides from the who le plan t o fB ulbophy llum odoratissim um