藏波羅花的化學(xué)成分研究

高燕萍，沈云亨，高燕霞，李 博，張衛(wèi)東*

1中國(guó)藥科大學(xué) 中藥學(xué)院，南京210009;2 第二軍醫(yī)大學(xué)藥學(xué)院 天然藥物化學(xué)教研室，上海200433

Introduction

Incarvillea younghusbandii，belonging to the genus Incarvillea (Bignoniaceae)，is a perennial herb native to the Qinghai and Tibet provinces of China［1］.As a Chinese folk medicine，this plant has been long used for the treatment of dizziness，anemia，and to stimulate lactation［2，3］.To date，phytochemical investigations on I.younghusbandii have just revealed the isolation of coumarins［4］，volatile oil［5］，phenolic glycosides［6］，and a dimeric cyclohexylethanoid［7］.In order to search for more interesting compounds with bioactivities，we investigated the chemical components of the titled plant.Ten compounds were obtained，all of which were isolated from I.younghusbandii for the first time.

Experimental

Apparatus and reagents

Melting points were determined on a RY-1 micro-melting point apparatus without correction. The ESI-MS were acquired on Agilent 1100 series mass spectrometer.NMR spectra were measured on a Bruker DRX-600 spectrometer with TMS as internal standard，operating at 600 MHz for1H and 150 MHz for13C NMR.Chemical shift (δ)was given in ppm and coupling constants in Hz. Column chromatographies (CC)were carried out on silica gel (200-300 mesh，Yantai，China)and sephadex LH-20 (GE Healthcare Bio-Sciences AB，Sweden)，and precoated silica GF254plates were used for TLC (Qingdao Haiyang Chemical Co.，Ltd.，China).

Plant material

The whole plants of I. younghusbandiii were collected in Zhongdian county，Yunnan province，P. R. China，in August 2010，and authenticated by Prof. Han-Ming Zhang of Second Military Medical University.A voucher specimen (No. 20100815)was deposited in the School of Pharmacy，Second Military Medical University.

Extraction and isolation

The air-dried whole plant (1.3 kg)were powered and extracted with 80% EtOH for four times and then partitioned with petroleum ether，EtOAc，and n-BuOH successively.The EtOAc-soluable extract was subjected to a silica gel column eluting with a gradient petroleum ether/Me2CO (30∶0→0∶1)to obtain frs.1-5.Fr.2 (1 g)were chromatographied on silica gel (200-300 mesh)eluting with a gradient of petroleum ether/Me2CO (10∶1)，and then purified by repeated column chromatography over Sephadex LH-20 (MeOH)，yielding compounds 3 (7. 5 mg)，5 (9. 0 mg)，7 (20. 0 mg)，9 (12.0 mg)，and 10 (12.5mg).Fr.3 (1.5 g)was applied to silica gel column chromatography and purified by PTLC (CHCl3:MeOH 15 ∶1)，to afford compounds 1 (8.0 mg)，2 (9.6 mg)，and 8 (27.0 mg).Fr.4 (0.5 g)，undertook to Sephadex LH-20 and combined with PTLC (CHCl3:MeOH 10 ∶1)，gave compounds 4 (15.0 mg)and 6 (17.5 mg).

Structural elucidation

Compound 1 C8H16O3;Colorless oil;ESI-MS:m/z 183［M+Na］+，159 ［M-H］-;1H NMR (CDCl3，600 MHz)δ:1.40-1.42 (2H，m，Hax-2 and Hax-6)，1.63-1.72 (2H，overlapped，m，Heq-2 and Heq-6)，1.63-1.72 (4H，overlapped，m，Hax-3 and Heq-3，Hax-5 and Heq-5 )，3.52 (1H，m，H-4)，1.69 (2H，t，J =7.2 Hz，H-1')，3.73 (2H，t，J = 6.6 Hz，H-2');13C NMR (CDCl3，150 MHz)δ:70.8 (C-1)，36.1 (C-2)，31.3 (C-3)，70.8 (C-4)，31.3 (C-5)，36.1 (C-6)，45.5 (C-1')，59.2 (C-2'). The NMR data was identical with those reported in the literature［8］，and elucidated as regyol.

Compound 2 C8H14O3;Colorless oil;ESI-MS:m/z 181［M+Na］+，157 ［M-H］-;1H NMR (CDCl3，600 MHz)δ:1.75 (2H，dt，J = 13.2，4.8 Hz，Hax-2，Hax-6)，2.10 (2H，m，Heq-2，Heq-6)，2.73 (2H，dt，J = 13.8，6.0 Hz，Hax-3，Hax-5)，2.22 (2H，m，Heq-3，Heq-5)，1.80 (2H，t，J = 6.0 Hz，H-7)，3.95(2H，t，J = 6.0 Hz，H-8);13C NMR (CDCl3，150 MHz)δ:70.5 (C-1)，36.7 (C-2)，37.0 (C-3)，212.9 (C-4)，37.0 (C-5)，36.7 (C-6)，41.5 (C-7)，59.5 (C-8). The NMR data were in accordance with those reported in the literature［9］. Therefore，compound 2 was characterized as cleroindicin B.

Compound 3 C11H21N;White powder;ESI m/z 168［M + H］+;1H NMR (CDCl3，600 MHz)δ:0.91(3H，d，J = 6.6 Hz，CH3-4)，1.01 (3H，d，J = 6.6 Hz，CH3-8)，1.18 (1H，m，H-4)，1.50 (1H，m，H-5)，1.62 (2H，m，Ha-6，Ha-7)，1.81 (1H，m，H-9)，1.98 (2H，m，Hb-6，Hb-7)，2.33 (1H，m，Hax-1)，2.44 (1H，brs，Hax-3)，2.76 (3H，s，N-Me)，2.91(1H，dd，J = 12.6，4.2 Hz，H-8)，3.23 (1H，d，J =10.8 Hz，Heq-3)，3.34 (1H，d，J = 10.8 Hz，Heq-1);13C NMR (CDCl3，150 MHz)δ:53.3 (C-1)，59.8(C-3)，30.5 (C-4)，45.6 (C-5)，27.2 (C-6)，31.8(C-7)，34.0 (C-8)，42.9 (C-9)，16.9 (4-CH3)，18.9 (8-CH3)，44.7 (N-CH3). The NMR data were in agreement with those reported in the literature［10］，and determined as β-skytanthine.

Compound 4 C21H32O12;White amorphous powder;ESI-MS:m/z 499［M + Na］+，475 ［M-H］-;1H NMR(CD3OD，600 MHz)δ:6.81 (lH，d，J = 8.2 Hz，H-5)，6.73 (1H，d，J = 2.0 Hz，H-2)，6.67 (1H，dd，J=8.2，2.0 Hz，H-6)，5.15 (lH，d，J = 16 Hz，H-1'')，4.29 (lH，d，J = 7.8 Hz，H-1')，3.81 (3H，s，Ar-OCH3)，2.81 (2H，t，J = 7.27 Hz，H-β)，1.25(3H，d，J = 6.2 Hz，H-6'')，4.08-3.24 (10H of ducose and rhamnose，m，H-α);13C NMR (CDCl3，150 MHz)δ:132.8 (C-1)，112.8 (C-2)，147.4 (C-3)，147.2 (C-4)，117.0 (C-5)，121.1 (C-6)，71.9 (Cα)，36.5 (C-β)，104.1 (C-1')，75.5 (C-2')，84.4(C-3')，71.1 (C-4')，77.7 (C-5')，62.6 (C-6')，102.6 (C-1'')，72.2 (C-2'')，72.1 (C-3'')，73.9(C-4'')，70.0 (C-5'')，17.9 (C-6'')，56.5 (3-OCH3). Compound 4 was determined as deacyl isomartynoside by comparison with the spectra data reported in the literature［11］.

Compound 5 C8H10O2;Colorless oil;ESI-MS:m/z 161［M+Na］+，137 ［M-H］-;1H NMR (CDCl3，600 MHz)δ:7.08 (2H，d，J = 8.4 Hz，H-2，H-6)，6.76(2H，d，J = 7.8 Hz，H-3，H-5)，3.81 (2H，t，J =6.6 Hz，H-α)，2.76 (2H，t，J = 6.6 Hz，H-β);13C NMR (CDCl3，150 MHz)δ:154.2 (C-1)，130.5 (C-4)，130.2 (C-3，C-5)，115.4 (C-2，C-6)，63.8 (Cα)，38.2 (C-β). The above data were identical with those reported in the literature［12］. Consequently，compound 5 was identified as tyrosol.

Compound 6 Amorphous solid，C14H24O8;ESI-MS:m/z 343［M + Na］+;1H NMR (600 MHz，CD3OD)δ:aglycone 4.15 (1 H，dt，J = 11.0，8.0 Hz，H-2"a)，3.86 (1H，dt，J = 11.0，8.0 Hz，H-2b ")，2.68(2H，ddd，J = 16.0，10.5，5.0 Hz，H-2ax，H-6ax)，2.31 (2H，dt，J = 16.0，10.5 Hz，H-2eq，H-6eq)，2.03 (2H，ddd，J = 16.0，10.5，5.0 Hz，H-3ax，H-5ax)，1.97 (2H，t，J = 8.0 Hz，H-l")，1.94 (2H，dt，J = 16.0，10.5 Hz，H-3eq，H-5eq);glucose 4.49(1H，d，J = 8.0 Hz，H-1')，3.92 (1H，dd，J =12.2，2.4 Hz，H-6'a)，3.71(1H，dd，J = 12.2，6.4 Hz，H-6'b)，3.49 (1H，t，J = 9.2 Hz，H-3')，3.47(1H，ddd，J = 9.2，6.4，2.4 Hz，H-5')，3.38 (1H，t，J = 9.2 Hz，H-4')，3.25 (1H，dd，J = 9.2，8.0 Hz，H-2');13C NMR (D2O，150 MHz)δ:219.5 (Cl)，103.0 (C-l')，76.7 (C-3')，76.5 (C-5')，73.8(C-2')，70.5 (C-4)，70.4 (C-4')，67.1 (C-2")，61.5(C-6')，40.6 (C-l")，37.1" (C-2，C-6)，36.5(C-3，C-5). By comparison the spectra data with those related in the literature［13］，compound 6 was elucidated as rengioside B.

Compound 7 C29H50O;White needles;mp.139-140° C;ESI-MS m/z 437.4 ［M + Na］+;13C NMR(CDCl3，150 MHz)δ:37.3 (C-1)，31.7 (C-2)，71.8(C-3)，42.3 (C-4)，140.8 (C-5)，121.7 (C-6)，31.7 (C-7)，31.9 (C-8)，50.2 (C-9)，36.5 (C-10)，21.1 (C-11)，39.8 (C-12)，42.3 (C-13)，56.8(C-14)，24.3 (C-15)，28.2 (C-16)，56.1 (C-17)，11.9 (C-18)，19.1 (C-19)，36.1 (C-20)，18.8 (C-21)，34.0 (C-22)，26.1 (C-23)，45.9 (C-24)，29.2(C-25)，19.8 (C-26)，19.4 (C-27)，23.1 (C-28)，12.0 (C-29). The above data were in accordance with those reported in the literature［14］. Consequently，compound 7 was identified as β-sitosterol.

Compound 8 C8H10O3;Colorless oil;ESI-MS:m/z 155［M + H］+，177 ［M + Na］+，153 ［M-H］-;1H NMR (CDCl3，600 MHz)δ:4.07 (1H，m，H-2a)，3.93 (1H，m，H-2b)，2.33 (1H，m，H-3a)，2.20(1H，m，H-3b)，6.77 (1H，d，J = 10.2 Hz，H-5)，6.00 (1H，d，J = 10.2 Hz，H-6)，2.78 (1H，dd，J =16.8，4.8 Hz，Hax-8)，2.60 (1H，dd，J = 16.5，5.4 Hz，Heq-8)，4.23 (1H，m，H-9);13C NMR (CDCl3，150 MHz)δ:66.2 (C-2)，39.5 (C-3)，75.4 (C-4)，148.3 (C-5)，128.5 (C-6)，197.1 (C-7)，40.1 (C-8)，81.4 (C-9). The NMR data were in accordance with those reported in the literature［15］，and determined as cleroindicin F.

Compound 9 C11H22NO;Colorless needles which showed a positive reaction to the Dragendorff reagent;mp.103-104 °C;ESI-MS:m/z 184 ［M + H］+;1H NMR (CDCl3，600 MHz)δ:2.68 (1H，ddd，J =2.0，6.0，12.0 Hz，H-1a)，1.56 (1H，t，J = 12.0 Hz，H-1b)，2.51 (1H，ddd，J =2.0，5.0，11.5 Hz，H-3a)，1.67 (1H，t，J = 12.0 Hz，H-3b)，2.08 (1H，m，H-4)，2.41 (1H，ddd，J = 2.0，6.0，12.0 Hz，H-5)，1.80 (1H，m，H-6a)，1.50 (1H，br q，J = 5.0，7.0，13.0 Hz H-6b)，4.31 (1H，td，J =2.0，6.5 Hz，H-7)，1.82 (1H，m，H-8)，1.93 (1H，pent，J = 6.0，6.0，12.0 Hz，H-9)，2.27 (3H，s，N-Me)，0.86 (3H，d，J = 6.6 Hz，Me-4)，1.02 (3H，d，J = 6.6 Hz，Me-8);13C NMR (CDCl3，150 MHz)δ:73.4 (C-7)，58.0 (C-1，C-3)，46.2 (N-Me)，45.8 (C-9)，42.3(C-8)，37.5 (C-5)，32.7 (C-6)，30.5 (C-4)，17.4(Me-8)，14.2 (Me-4). Comparing NMR data with those reported in the literature［16］，compound 9 was determined as incarvilline.

Compound 10 C8H8O4;White powder;ESI-MS:m/z 167［M-H］-;1H NMR (CD3OD，600 MHz)δ:7.55(2H，s，H-2 and H-6)，6.82 (1H，d，J = 7.8 Hz，H-5)，3.88 (3H，s，3-OCH3);13C NMR (CD3OD，150 MHz)δ:123.0 (C-1)，113.9 (C-2)，152.5 (C-3)，148.6 (C-4)，115.8 (C-5)，125.2 (C-6)，170.0 (C-7)，56.4 (3-OCH3). The above data were in agreement with those dealt with in the literature［17］，and identified as 3-O-methyl-4-hydroxy benzoic acid.

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