观光木酚性成分研究

黄 鹰，常睿洁，金慧子*，张卫东，2*

1上海交通大学药学院，上海200240;2上海第二军医大学药学院，上海200433

Introduction

Tsoongiodendron odorum Chun(Magnoliaceae)is an evergreen tree，mainly distributed in south of China，such as Jiangxi，Guangdong，Yunnan and so on［1］.The stem barks and root barks of this plant have been used to treat cancer by the local people for a long time.Up to now，only a few chemical constituents have been isolated from this plant［2-4］.In this study on the twigs of T.odorum，eleven phenolic constituents were isolated and identified as p-hydroxybenzaldehyde(1)，vanillin (2)，1，3，5-trimethoxybenzene(3)，4-hydroxy-3，5-dimethoxy-benzaldehyde(4)，vanillic acid(5)，threo-2， 3-bis-(4-hydroxy-3-methoxyphenyl)-3-methoxy-propanol (6)，erythro-2，3-bis-(4-hydroxy-3-methoxyphenyl)-3-methoxy-propanol(7)，syringic acid(8)，3-hydroxy-1-(4-hydroxy-3，5-dimethoxyphenyl)-1-propanone(9)，p-hydroxyl phenethanol(10)，and C-veratroylglycol (11).Compounds 1-4 and 6-11 were isolated from this plant for the first time.

Experimental

General procedures

All melting points were determined on an RY-1 micromelting point apparatus(uncorrected).The ESIMS were measured on an Agilent 1100 series mass spectrometer.NMR spectra were measured on a Bruker DRX-500 spectrometer，operating at 500 MHz for1H and 125 MHz for13C NMR.Chemical shift(δ)were given in ppm relative to TMS as internal standard and coupling constants in Hz.Silica gel(200-300 mesh， Yantai，China)was used for column chromatography，and precoated silica GF254plates were used for TLC (Qingdao Haiyang Chemical Co.，Ltd.，China).Semipreparative HPLC was obtained on a Shimadzu LC-6AD series equipped with an SPD-20 spectrophotometer using a Zorbax-SB-C185 μm(9.4×250 nm，ID).

Plant material

The twigs of T.odorum were collected from Jiangxi Province of China in July，2008 and identified by Prof.Huang Bao-kang，Department of Pharmacognosy，School of Pharmacy，Second Military Medical University.A voucher specimen was deposited at School of Pharmacy，Shanghai Jiao Tong University.

Extraction and Isolation

The air-dried and powdered twigs of T.odorum(10.0 kg)were extracted with 80%EtOH for three times at room temperature.The solvents were concentrated under vacuum to yield a residue(525.0 g).The residue was dissolved in water(5.0 L)and extracted successively with petroleum ether(5.0 L×10)，ethyl acetate (5.0 L×10)and n-butanol(5.0 L×10)to give corresponding fractions A(38.5 g)，B(83.6 g)，and C (185.5 g).Fraction B was subjected to silica gel column chromatography with a step gradient of CH2Cl2-MeOH(100∶1-0∶100)to give 12 fractions B1—B12.Fraction B2 was separated on silica gel column eluted by CH2Cl2-MeOH(20∶1-1∶1)to give 8 fractions B21—B28.Fraction B21 was subjected repeatedly to silica gel column chromatography and further purified successively through semi-HPLC to yield compounds 1 (7.0 mg)，2(10.0 mg)，3(60.0 mg)，and 4(32.5 mg).Similarly，fraction B22—B24 were purified in the same manner to yield compounds 5(30.0 mg)，6(1.5 mg)，7(1.5 mg)，8(4.5 mg)，9(6.2 mg)(5-9 from B22)，10(12.0 mg，from B23)，and 11(7.5 mg，from B24).

Results and Discussion

p-Hydroxybenzaldehyde(1)C7H6O2，white needle crystal(CH3OH)，mp.110～112℃;ESI-MS m/z:121［M–H］–;1H NMR(CD3OD)δ:9.76(1H，s，CHO-1)，7.77(2H，d，J=8.5 Hz，H-2，6)，6.91 (2H，d，J=8.5 Hz，H-3，5);13C NMR(CD3OD)δ: 192.8(CHO-1)，165.2(C-4)，133.4(C-2，6)，130.3(C-1)，116.9(C-3，5).The NMR and MS data were in accordance with those reported in the literature［5］，and compound 1 was identified as p-hydroxybenzaldehyde.

Vanillin (2) C8H8O3，white needle crystal (CH3OH)，mp.68～71℃;ESI-MS m/z:151［M–H］–;1H NMR(CDCl3)δ:9.83(1H，s，CHO-1)，7.43(2H，br s，H-2，6)，7.04(1H，d，J=7.0 Hz，H-5)，3.97(3H，s，CH3O-3);13C NMR(CDCl3)δ: 190.7(CHO-1)，151.7(C-4)，147.2(C-3)，129.9 (C-1)，127.3(C-6)，114.4(C-5)，108.9(C-2)，56.1(CH3O-3).The NMR and MS data were in accordance with those reported in the literature［6］，and compound 2 was identified as vanillin.

1，3，5-Trimethoxybenzene(3) C9H12O3，yellow needle crystal(CH2Cl2)，mp.53～55℃;ESI-MS m/ z:191［M+Na］+;1H NMR(CDCl3)δ:5.85(3H，s，H-2，4，6)，3.81(9H，s，CH3O-1，3，5);13C NMR (CDCl3)δ:157.3(C-1，3，5)，107.4(C-2，4，6)，56.4(CH3O-1，3，5).The NMR and MS data of 3 were in accordance with those reported in the literature［7］.

4-Hydroxy-3，5-dimethoxy-benzaldehyde(4)

C9H10O4，light yellow solid，mp.110～113℃;ESI-MS m/z:181［M–H］–;1H NMR(CDCl3)δ:9.81 (1H，s，CHO-1)，7.15(2H，s，H-2，6)，3.96(6H，s，CH3O-3，5);13C NMR(CDCl3)δ:190.7(CHO-1)，147.4(C-3，5)，140.9(C-4)，128.3(C-1)，106.7 (C-2，6)，56.4(CH3O-3，5).The NMR and MS data were consistent with those reported in the literature［8］. Vanillic acid(5) C8H8O4，white needle crystal (CH3OH)，mp.159-161℃;ESI-MS m/z:167［M–H］–;1H NMR(CD3OD)δ:7.55(2H，m，H-2，6)，6.84(1H，d，J=8.5 Hz，H-5)，3.89(3H，s，CH3O-3);13C NMR(CD3OD)δ:170.0(COOH-1)，152.7 (C-4)，148.7(C-3)，125.3(C-6)，123.1(C-1)，115.8(C-5)，113.8(C-2)，56.4(CH3O-3).The NMR and MS data were in accordance with those reported in the literature［9］，and compound 5 was identified as vinillic acid.

threo-2，3-Bis-(4-hydroxy-3-methoxyphenyl)-3-methoxy-propanol(6) C18H22O6，white oil;ESI-MS m/z:333［M–H］–;1H NMR(CD3OD)δ:6.46-6.72(6H，m，ArH)，4.30(1H，d，J=8.5 Hz，H-7)，4.06(1H，m，H-9a)，3.87(1H，m，H-9b)，3.75，3.67(each 3H，s，CH3O-3，3')，3.21(3H，s，CH3O-7)，3.00(1H，m，H-8);13C NMR(CD3OD)δ: 148.6，148.3(C-3，3')，146.9，146.2(C-4，4')，133.2，132.6(C-1，1')，123.2，121.5(C-6，6')，115.7，115.5，114.7，112.4(C-2，2'，5，5')，87.4 (C-7)，64.9(C-9)，57.0，56.5，56.3(CH3O-3，3'，7)，56.1(C-8).The NMR and MS data were consistent with those reported in the literature［10］.

erythro-2，3-Bis-(4-hydroxy-3-methoxyphenyl)-3-methoxy-propanol(7)C18H22O6，white oil;ESI-MS m/z:333［M–H］–;1H NMR(CD3OD)δ:6.46-6.72(6H，m，ArH)，4.47(1H，d，J=5.5 Hz，H-7)，3.89(2H，m，H-9)，3.68，3.65(each 3H，s，CH3O-3，3')，3.15(3H，s，CH3O-7)，2.85(1H，m，H-8);13C NMR(CD3OD)δ:148.5，148.3(C-3，3')，146.8，146.0(C-4，4')，132.6，132.3(C-1，1')，122.6，121.3 (C-6，6')，115.6，115.5，114.4，118.8(C-2，2'，5，5')，84.9(C-7)，64.4(C-9)，56.8，56.4，56.2(CH3O-3，3'，7)，56.0(C-8).The NMR and MS data of 7 were in accordance with those reported in the literature［11］.

Syringic acid(8) C9H10O5，white solid，mp.187～190℃;ESI-MS m/z:197［M– H］–;1H NMR (CD3OD)δ:7.33(2H，s，H-2，6)，3.88(6H，s，CH3O-3，5);13C NMR(CD3OD)δ:170.0(COOH-1)，148.7(C-3，5)，141.0(C-4)，123.9(C-1)，108.3(C-2，6)，56.8(CH3O-3，5).The NMR and MS data were in accordance with those reported in the literature［12］.

3-Hydroxy-1-(4-hydroxy-3，5-dimethoxyphenyl)-1-propanone(9)C11H14O5，light yellow solid，mp.78～81℃;ESI-MS m/z:225［M– H］–;1H NMR (CD3OD)δ:7.30(2H，s，H-2，6)，3.94(2H，t，J= 6.0 Hz，H-9)，3.87(6H，s，CH3O-3，5)，3.15(2H，t，J=6.0 Hz，H-8);13C NMR(CD3OD)δ:199.6(C-7)，149.7(C-3，5)，138.0(C-4)，127.0(C-1)，107.6(C-2，6)，59.2(C-9)，56.8(CH3O-3，5)，41.5(C-8).The NMR and MS data were consistent with those reported in the literature［13］.

p-Hydroxyl phenethanol(10) C8H10O2，light yellow oil;ESI-MS m/z:137［M– H］–;1H NMR (CD3OD)δ:6.95(2H，d，J=8.0 Hz，H-2，6)，6.62 (2H，d，J=8.0 Hz，H-3，5)，3.60(2H，t，J=7.0 Hz，H-8)，2.64(2H，t，J=7.0 Hz，H-7);13C NMR (CD3OD)δ:156.9(C-4)，131.2(C-1)，131.0(C-2，6)，116.3(C-3，5)，64.7(C-8)，39.5(C-7).The NMR and MS data were in accordance with those reported in the literature［14］.

C-veratroylglycol(11) C10H12O5，light yellow oil; ESI-MS m/z:211［M–H］–;1H NMR(CD3OD)δ: 7.50(2H，m，H-2，6)，6.80(1H，d，J=8.0 Hz，H-5)，5.04(1H，m，H-8)，3.83(3H，s，CH3O-3)，3.80 (1H，m，H-9a)，3.66(1H，m，H-9b);13C NMR (CD3OD)δ:199.7(C-7)，152.3(C-4)，149.4(C-3)，127.3(C-1)，125.3(C-6)，116.1(C-2)，112.6 (C-5)，75.7(C-8)，66.4(C-9)，56.5(CH3O-3).

The NMR and MS data were in accordance with those reported in the literature［15］.

1 Editorial committee of flora of china.Flora of China.Beijing: Science Press，1999，30:194.

2 Song XK，Tu PF，Wu LJ，et al.A new sesquiterpene lactone from Tsoongiodendron odorum Chun.J Asian Nat Prod Res，2001，3:285-291.

3 Wang BG，Hong X，Li L，et al.Chemical constituents of two Chinese Magnoliaceae plants，Tsoongiodendron odorum and Maglietiastrum sinicum，and their inhibition of platelet aggregation.Planta Med，2000，66:511-515.

4 Song XK，Wu LJ，Tu PF.Studies on bioactive constituents in bark of Tsoongiodendron odorum.Chin Tradit Herb Drugs，2002，33:676-678.

5 Li M，Guo SX，Wang CL，et al.Studies on chemical constituents of the Tubers of Gymnadenia conopsea.Chin Pharm J，2008，43:409-412.

6 Ren YL，Tang QR，Zhang Z，et al.Chemical constituents from Tinospora sinensis.Nat Prod Res Dev(天然产物研究与开发)，2008，20:278-279.

7 Hu YM，Wang XR，Ye ZW，et al.A new approach for synthesis and characterization of 1，3，5-trimethoxybenzene.Chin J Synth Chem，1999，7:191-193.

8 Liu DL，Pang FG，Zhang JX，et al.Studies on the chemical constituents of Bulbophyllum odoratissimum Lindl.Chin J Med Chem，2005，15:103-107.

9 Akiyo S，Maksut C，Takashi M.Hydroxybenzoic acid from Boreava orientalis.Phytochemistry，1995，40:257-261.

10 Hsiao JJ，Chiang HC.Lignans from the wood of Aralia bipinnata.Phytochemistry，1995，39:899-902.

11 Alder E，Brunow G，Lundquist K.Investigation of the acid-catalysed alkylation of lignins by means of NMR spectroscopic methods.Holzforschung，1987，41:199-207.

12 Deng MC，Jiao W，Dong WW，et al.Study on the chemical constituents of Uncaria rhynchophylla.Nat Prod Res Dev(天然产物研究与开发)，2009，21:242-245.

13 Ma ZJ，Zhao ZJ.Study on chemical constituents from stem barks of Fraxinus paxiana.China J Chin Mat Med，2008，33:1990-1993.

14 Qiong MX，Yan LL，Xiao RL，et al.Two new phenylglycol derivatives isolated from Syringa reticulate var.mandshurica and their antifungal activities.Chem Pharm Bull，2009，57: 863-866.

15 Jones L，Bartholomew B，Latif Z，et al.Constituents of Cassia laevigata.Fitoterapia，2000，71:580-583.