291 lisicamina

8/11/2019 291 lisicamina

1/8

The Constituents of Lindera glauca

Yuh-Chwen Chang

a,b

( ), Fang-Rong Chang

b

( ) and Yang-Chang Wu*

b

( )aDepartment of Chemical Engineering, Kao Yuan Institute of Technology, Kaohsiung county, Taiwan, R.O.C.

bGraduate Institute of Natural Products, Kaohsiung Medical University, Kaohsiung, Taiwan, R.O.C.

Twenty-eight compounds including seven alkaloids, (+)-3-chloro-N-formylnornantenine (1), (+)-N-

formylnornantenine (2), (+)-boldine (3), (+)-norboldine (4), (-)-norboldine (5), lycicamine (6), and tetrahydro-

berber ine (7); four flavonoids, kaempferol (8), kaempferol-3-O-arabinoside (9), quercetin (10), and

quercetin-3-O-rhamnoside (11); one butanolide, akolactone A (12); onep-quinone, 2,6-dimethoxy-p-quinone

(13); one cyclohex-2-en-1-one, blumenol A (14); six benzenoids, methylparaben (15),p-hydroxybenzoic acid

(16), vanillic acid (17), syringic acid (18), 3,4,5-trimethoxybenzoic acid (19), and 3-(3,4-dihydroxyphenyl)

propionic acid (20); one diterpene, phytol (21); one triterpene, squalene (22); six steroids, -sitosterol (23),

-sitostenone (24), stigmasta-4,22-dien-3-one (25), 6 -hydroxy- -sitostenone (26), 6 -hydroxystigmasterone

(27), and -sitosteryl-D-glucoside (28) were isolated from the aerial part ofLindera glauca. These compoundswere characterized and identified by physical and spectral method. All compounds were isolated for the first

time from this plant. Among them, (+)-3-chloro-N-formylnornantenine (1) is a new one.

INTRODUCTION

Lindera glauca Sieb.& Zucc.(Lauraceae) is a small de-

ciduous tree growing in the forests at low altitudes in Japan,

China and Taiwan.1

Previously, three alkaloids, nineteen fatty

acids, eight monoterpenes, five sesquiterpenes, two naph-

thalenes, and eleven butanolideshave been reported from this

plant.2-8 As part of our continuing investigation of the phyto-

chemical and bioactive compounds of Formosan Lauraceous

plants, the methanol extract of this plant was subjected to sol-

vent partition and chromatographic separation to characterize

twenty-eight pure compounds. Among them, 1is a new com-

pound, and known compounds 2~28were isolated for the first

time from this species. The present paper deals with the isola-

tion and characterization of the isolated components.

RESULTS AND DISCUSSION

A methanolic extract ofL. glaucawas concentrated to

obtain a residue and then partitioned between CHCl3and H2O.

The aqueous layer was partitioned with n-BuOH. Each layer

was concentrated and subjected to chromatography. Fifteen

compounds including five alkaloids, (+)-3-chloro-N-formyl-

nornantenine (1), (+)-N-formylnornantenine9(2), (+)-nor -

boldine10

(4), (-)-norboldine11,12

(5), and lysicamine13

(6); one

butanolide, akolac tone A (12); one benzenoid, methyl-

paraben14

(15); one diterpene, phytol (21); one triterpene,

squalene (22); and five steroids, -sitosterol13,15 (23), -

sitostenone13 (24), stigmasta-4,22-dien-3-one13 (25), 6 -

hydroxy- -sitostenone13 (26), 6 -hydroxystigmasterone13

(27), and -sitosteryl-D-glucoside13

(28) were obtained from

Journal of the Chinese Chemical Society, 2000, 47, 373-380 373


2/8

374 J. Chin. Chem. Soc., Vol. 47, No. 2, 2000 Chang et al.


3/8

the chloroform layer. Thirteen compounds including two al-

kaloids, (+)-boldine11,12

(3) and tetrahydroberberine16

(7);

four flavonoids, kaempferol17 (8), kaempferol-3-O-

arabinoside18 (9), quercetin17 (10 ), and quercetin-3-O-

rhamnoside19 (11 ); one p-quinone, 2,6-dimethoxy-p-

quinone20 (13); one cyclohex-2-en-1-one, blumenol A21,22

(14); five benzenoids,p-hydroxybenzoic acid14

(16), vanillic

acid14

(17), syringic acid14

(18), 3,4,5-trimethoxybenzoic

acid14 (19), and 3-(3,4-dihydroxyphenyl) propionic acid23

(20) were obtained from the n-Butanol layer. Among them 1is

a new compound, and others are known compounds which

were isolated for the first time from this plant and character-

ized by comparison of their physical and spectral data (UV,

IR, NMR and MS) with values previously reported in the liter-

ature.

The alkaloid 1was recrystallized from methanol as col-

orless needles. Its molecular formula was established as

C20H18O5NCl by HREI mass spectrometry m/z M+

(found

387.0867, calcd 387.0874), with a mp over 300 C. The IR

spectrum with peaks at 1660 and 1620 cm-1, was suggestive of

an amidic functional group. The UV absorptions showed max

at 272sh, 281, 302, and 312sh nm, indicating alkaloid 1pos-

sessed a 1, 2, 3, 9, 10-substituted aporphine24

. EI-MS spec-

trum showed that one chlorine is bonded to alkaloid 1with a

spectrum of 3 to 1 ratio at m/z (rel. int %): 387(23, M+)/389;

351(52, [M-Cl]+)/353; 329(100)/331. The molecular ion m/z

387(23, M+)/389, and base peak m/z329(100)/331 are due to

loss of (CH2-N-CHO+H) from the molecular ion. It followed

that the amidic function was in the shape of an N-formyl

group. Moreover, the two species were actually present in so-

lution, due to isomerism about the amidic bond could be im -

mediately derived from the 1H-NMR spectrum. Broad down-

field singlets at 8.25 and 8.36 represented theN-formyl pro-

ton. The integrals of these peaks indicated that the isomers

were present in a ratio of 2:1.

The spectra for the two isomers could be clearly differ-

entiated, even though the two isomers could not be separated.

While the differences in chemical shifts were minimal in the

case of the methoxyl and methylenedioxy signals, they were

clearly noticeable for the aromatic protons, with signals at

6.78 (H-8) and 7.91 (H-11) for the major isomer 1a, and at

6.76 and 7.92 for the minor isomer 1b . The divergence be-

tween the two isomers was quite prominent in the aliphatic

range. Some of the more salient differences in chemical shifts

occurred in the resonances for the hydrogens bonded to C-5,

C-6a and C-7. For the major isomer 1a, the signals for H-6a

The Constituents ofLindera glauca J. Chin. Chem. Soc., Vol. 47, No. 2, 2000 375


4/8

and H-7 were downfield at 4.91 and 2.97, respectively. On

the other hand, for 1b, these two protons are relatively upfield

at 4.45 and 2.75, respectively. For both isomers, the H-7

signals were characterized by small coupling constants withH-6a (ca. 4.0 Hz), but the H-7 signals were denoted by large

constants (ca. 14.0 Hz).

The disparity in geometry between the two isomers also

came to the fore when NOESY measurements were obtained.

In particular, the proximity of the formyl proton to H-5 (

3.86) in the major isomer and to H-6a ( 4.45) in the minor iso-

mer was clearly evident. With both isomers, a strong nOe

could be observed between H-7 and H-8 (see Fig. 1). A simi-

lar observation for alkaloid 2has been found in the literature.9

In view of alkaloid 2, the H-3 signal disappeared in

those of 1. It strongly indicates that the chlorine atom was sub-

stituted at 3-position. Since neither chlorine gas nor HCl was

even used during extraction or chromatography, an artifact is

excluded. On the basis of the result mentioned above, the

structure was thus determined as (+)-3-chloro-N-formyl-

nornantenine. In addition, the biological activities of these

compounds are currently under investigation.

EXPERIMENTAL SECTION

General Methods

Uncorrected melting points were determined on a

Mel-Temp II apparatus. Optical rotation was taken on a Jasco

DIP-370 digital polarimeter. Ultraviolet (UV) and infrared

(IR) spectra were obtained using a Hitachi U-2000 spectro-

photometer and a Hitachi 260-30 spectrophotometer, respec-

tively. NMR spectra were recorded with a Varian Gemini

NMR Spectrometer (200 MHz) and a Varian Unity Plus NMR

Spectrometer (400 MHz) using TMS as an internal standard.

Mass Spectra (MS) were recorded on a JEOL JMS-HX 110

mass spectrometer. Active charcoal (Wako) and silica gel 60

(E. Merck, 230-400 mesh) were used for open column chro-

matography and precoated silica gel plates (E. Merck,

Kieselgel 60 F-254, 0.25 mm) were used for preparative TLC.

Plant MaterialL. glauca Sieb. Zucc.(Lauraceae) was collected from

Taipei county, Taiwan in July 1996. A voucher specimen is on

deposit in the Graduate Institute of Natural Products,

Kaohsiung Medical University, Kaohsiung, Taiwan, Republic

of China.

Extraction and Separation

Air-dried, aerial parts (9.8 Kg) of L. glauca were ex-

tracted repea tedly with MeOH at room temperature. The

methanolic extracts were concentrated and partitioned be-

tween chloroform and water to form a chloroform layer and an

aqueous layer. The chloroform layer was concentrated to

leave a brownish viscous residue (8.8 g). The residue was

placed on a silica gel column and eluted with CHCl3 which

was gradually enriched with MeOH to afford eleven fractions.

Fr.2 (2.1 g) eluted with a gradient of n-hexane/CHCl3/EtOAC

was separated using silica gel CC and prep. TLC and gave

(+)-3-chloro-N-formylnornantenine (1) (3 mg), (+)-N-

formylnornantenine (2)(5 mg),akolactone A (12) (3 mg),

phytol (21) (20 mg) and squalene (22) (20 mg), respectively.

Fr.5 (1.4 g) eluted with CHCl3 was isolated using repeatedly

silica gel CC and prep. TLC (CHCl3) and gave -sitosterol

(23) (50 mg). (+)-norboldine (4) (10 mg), (-)-norboldine (5) (4

mg)andmethylparaben (15) (10 mg) were obtained from Fr.7

(1.3 g) by means of repea tedly silica gel CC eluting with

CHCl3-MeOH (20:1). Fr.8 (0.6 g) eluted with CHCl3-MeOH

(15:1) was further separated using silica gel CC and prep.

TLC and gave lysicamine (6) (5 mg). Fr.10 (0.8 g) eluted with

CHCl3-MeOH (10:1) was separated using silica gel CC and

prep. TLC and gave -si tostenone (24) (10 mg), stigmasta-

4,22-dien-3-one (25)(15 mg), 6 -hydroxy- -sitostenone (26)

(10 mg) and 6 -hydroxystigmasterone (27) (10 mg), respec-

tively. Fr.11(1.1 g) eluted with CHCl3-MeOH (8:1) was fur -

ther separated using silica gel CC and gave -sitosteryl-D-glucoside (28) (30 mg).

The aqueous layer was partitioned with n-Butanol to

give a n-Butanol layer and an aqueous layer. The n-Butanol

layer was concentrated (4.8 g) and chromatographed over sil-

ica gel using CHCl3/EtOAC/MeOH as an eluent to produce

ten fractions. Fr.3 (0.8 g) eluted with CHCl3was further puri-

fied in silica gel column using the same solvent to obtain com-

pound 2,6-dimethoxy-p-quinone (13) (10 mg). Fr.4 (2.3 g)

eluted with CHCl3-MeOH (10:1) was further separated using

silica gel CC and prep. TLC and gave kaempferol (8) (10 mg),

quercetin-3-O-rhamnoside (11) (50 mg); blumenol A (14) (30

mg),p-hydroxybenzoic acid (16) (20 mg), vanillic acid (17)


Fig. 1. NOESY correlations for 1.


5/8


6/8


7/8


8/8

2113-M-037-009)

Received August 4, 1999.

Key Words

Lindera glauca; Alkaloids; Flavonoids;

Benzenoids; Diterpenes; Triterpens; Steroids;

(+)-3-Chloro-N-formylnornantenine.

REFERENCES

1. Chang, C. E. In Flora of Taiwan; Li, H. L.; Liu, T. S.;

Huang, T. C.; Koyama, T.; Devol, C. E. Eds.; Epoch Pub-

lishing Co.; Taipei, 1976, Vol. II, p. 429.

2. Mutsuo, K.; Masahiko, S. J. Nat. Prod.1984, 47, 1066.

3. Nii, H.; Furukawa, K.; Iwakiri, M. Nippon Nogei Kagaku

Kaishi 1983, 57, 725.

4. Nii, H.; Furukawa, K.; Iwakiri, M. Nippon Nogei Kagaku

Kaishi 1983, 57, 733.

5. Nii, H.; Hanafusa, M.; Furukawa, K. Yukagakyu1983, 32,

277.

6. Komae, H.; Hayashi, N. Phytochemistry 1972, 11, 853.

7. Sasaki, T.; Haga, K.; Kaneko, R. Phytochemistry1994,

36, 949.

8. Hatsura, S.; Takayuki, S.; Satoshi, W.; Kazuo, H.; Ryohei,

K. Phytochemistry 1995,40, 1175.

9. Bamrung, T.; Tharadol, P.; Mataya, P. J. Nat. Prod.1989,

52, 652.

10. Borthakur, N.; Rastagi, R. C. Phytochemistry 1979, 18,

910.

11. Hema, U.; Bhakuni, D. S.; Dhar, M. M. Phytochemistry

1972, 11, 3057.12. Torrero, Y. M.; Docon, M. P. J. Nat. Prod.1990, 53, 503.

13. Chen, C. Y.; Chang, F. R.; Wu, Y. C. J. Chin. Chem. Soc.

1997, 44, 313.

14. Chen, C. Y.; Chang, F. R.; Teng, C. M.; Wu, Y. C. J. Chin.

Chem. Soc.1999,46, 77.

15. Chen, K. S.; Chang, F. R.; Chia, Y. C.; Wu, T. S.; Wu, Y. C.

J. Chin. Chem. Soc. 1998, 45, 103.

16. Malhotra, S.; Taneja, S. C.; Dhar, K, L. Phytochemistry

1989, 28, 1998.

17. Shimizu, M.; Ito, T.; Terashima, S.; Hayashi, T.; Arisawa,

M.; Morita, N.; Kurokawa, S.; Ito, K.; Hashimoto, Y.

Phytochemistry 1984, 23, 1885.

18.Nakano, K.; Takatani, M.; Tomimatsu, T.; Nohara , T.


19. Markham, R.; Ternai, B.; Stanley, R.; Geiger, H.; Mabry,

T. J. Tetrahedron1978, 34, 1389.

20. Kokpol, U.; Warinthom, C.; Vallapa, C.; Marc, B.;

Cunningham, G. N. Phytochemistry 1993, 33, 1129.

21. Bhakuni, D. S.; Josh, P. P.; Uprety, H.; Kapil, R. S.


22. Galbraith, M. N.; Horn, D. H. S. J. Chem. Soc ., Chem.

Comm., 1972, 113.

23. Meselhy, M. R.; Nakamura, N.; Hattori, M. Chem. Pharm.

Bull.1977, 45, 888.

24. Shamma, M. The Isoquinoline Alkaloids, Academic

Press: New York, 1972;Vol. 25, pp.194-314.


291 lisicamina

Documents