Cô lập của SESQUITERPENOIDS từ thân rễ của CURCUMA AROMATICA SALISB Việt Nam

Cô lập của SESQUITERPENOIDS từ thân rễ của CURCUMA AROMATICA SALISB Việt Nam.

Journal of Chemistry, Vol 38, No 4, P 96 - 99, 2000

ISOLATION OF SESQUITERPENOIDS FROM THE RHIROMES OF

VIETNAMESE CURCUMA AROMATICA SALISB

Received 5-6-2000

Phan Minh Giang, Phan Tong Son

College of Natural Science, Vietnam National University, Hanoi

Summary

The petroleum ether extract from the rhizomes of Curcuma aromatica Salisb growing in Vietnam was subjected to repeated flash chromatography on silica gel to give six oxygenated

sesquiterpenes furanodiene (1), furanodienone (2), curzerenone (3), germacrone (4), curcumenone (5) and zederone (6) together with three sesquiterpene hydrocarbons -humulene,

-selinene, -selinene

INTRODUCTION

The rhizomes of C aromatica Salisb.,

Zingiberaceae, have been used medicinally in

China, Japan and Southeast Asia [1] As it

contains bioactive principles, the chemical

composition of C aromatica have been

investigated extensively and it is recognized to

be a rich source of terpenoids However, these

investigations were focused on the plant

growing in India, China and Japan [2-12], little

is known about that growing in Vietnam

Hitherto, only the essential oil from C

aromatica rhizomes collected in Northern

Vietnam has been analysed [13, 14]

Curzerenone, germacrone as the major

constituents together with minor sesquiterpene

hydrocarbons and monoterpenoids were

identified [14]

In a previous paper [15], we reported the

results of our investigation on the composition

of the extracts from the rhizomes of C

aromatica Salisb growing in Vietnam by

means of GC, GC-MS and NMR methods This

paper goes into details of the isolation and

identification of the sesquiterpenoid

constituents

RESULTS AND DISCUSSION Repeated chromatography on silica gel of the petroleum ether soluble-fraction of an

EtOH extract of C aromatica rhizomes

gave three sesquiterpene hydrocarbons ( -humulene, -selinene and -selinene) and six

oxygenated sesquiterpenes (furanodiene (1), furanodienone (2), curzerenone (3), germacrone (4), curcumenone (5) and zederone (6)) Of

these, the hydrocarbons were identified by comparison of their MS and 1H-NMR spectra with those of authentic samples together with the relative retention indices (RRI) measured

on GC capillary column CP Sil5 CB

Furanodiene (1) [16], furanodienone (2) [17] and curzerenone (3) [18] were first found

in C zedoaria, Zingiberaceae We also isolated

the interesting sesquiterpenoid germacrone (4)

which also was found in many Curcuma

species The spectral data (MS, 1H- and 13

C-NMR) of the compounds 1 - 4 are in full

agreement with those isolated by us from C aff

aeruginosa Roxb., Zingiberaceae [19] Of the

two main constituents 3 and 4 of this extract (GC: together 30.7%), curzerenone (3) has been

found to be active against Mycobacterium tuberculosis H37RV [14]

Curcumenone (5) was isolated as colorless oil from a more polar fraction; 5 has also been

isolated from C zedoaria [20], C longa [21] and C aromatica of Japan [3] Biogenetically, this

seco-guaiane-type dicetone may be formed from germacrone via a guaiane-type cationic intermediate [20]

The IR spectrum of compound 6 (C15H18O3,

from HR-MS) exhibited absorption bands due

to an , -unsaturated cetone (1674 cm-1), and

an epoxide ring (846 cm-1), no hydroxyl or

other carbonyl absorption bands were observed

The 13C-NMR and DEPT spectra of 6 indicated

the presence of 15 C, including a cetonic

carbonyl [ 192.1 ppm (s)], two oxygenated

carbons [ 63.9 (s) and 66.5 ppm (d)], 6

olefinic carbons (4s and 2d), three methylene

and three methyl groups The 1H-NMR

spectrum showed characteristic signals for a

trisubstituted furan ring [ 7.06 (s), -H; 2.09

ppm (s), -CH3], and a vinylic proton [5.46

ppm (dd, J = 4 Hz, 12 Hz)] The chemical shift

and coupling constants of this vinylic proton

suggested that the cetonic carbonyl could only

be in conjugation with the furan ring The

presence of an epoxy function was supported by

the signals at 1.32 (s, 4-Me) and 3.79 (s, H-5)

in 1H-NMR spectrum as well as two carbon

signals at 63.9 (s) and 66.5 (d) in 13C-NMR

spectrum The other major feature of the 1

H-NMR spectrum of 6 was an AB system (2H)

with doublets centered at 3.66 and 3.73 ppm

(J = 16 Hz) (2 H-9) Thus on the basis of

spectral data, a furanogermacrane skeleton for

6 was suggested The complete 1H-1H COSY and HMQC spectral data confirmed the

structure of zederone (6) Zederone was

reported to be found in C zedoaria [22, 23]

The relative stereochemistry of 6 was

established by a NOESY experiment to be identical with the revised stereochemical

structure proposed by I Kitagawa et al [24]

Figure 1: Important NOESY interactions of 6

EXPERIMENTAL

General procedures

All melting points are uncorrected Optical rotation was obtained on a Polartronic D

CH3 H

CH 3 O

O

H

H

H H H

H H

O

10 6

9 8

12

13 15

14

5 4 3 2

1

O

O

O 6

5

13

12 11

14 4 3

2

110

9 8 7 15

3

O O

2 1

O

O

O

O

H

(Schmidt + Haensch) H-NMR (CDCl3):

Bruker AM 400 13 C-NMR (CDCl3): Bruker AC

200 with DEPT programme For zederone (6)

all NMR spectra were taken on a Bruker AMX

600 IR (CCl4): Perkin-Elmer 881 HR-MS:

Finigan MAT 95 SQ GC-EIMS:

Hewlett-Packard 5890 II with a HP-1 column (25 m x

0.2 mm i.d., film thickness 0.5 µm) combined

with MSD HP 5971A, carrier gas He GC:

Packard 439 with a CP Sil 5 CB column (25 m x

0.25 mm i.d., film thickness 0.39 µm); carrier

gas: N2; detector: FID; plotter: Shimadzu

C-R6A Chromatopack; temperature programme:

60 - 220oC, 5oC/min; injector temperature:

270oC; detector temperature: 300oC FC: silica

gel ICN Biomedicals (32-63 µm) TLC:

DC-Alufolien 60 F254 No (Merck 5554)

Visualisation: UV light 254 nm,

anisalde-hyde/H2SO4

Extraction and Isolation

The dried rhizomes of C aromatica (1kg)

collected in Socson, Hanoi (Vietnam) were

powdered and extracted with EtOH 96% by

percolation at room temperature The combined

extract was partially concentrated under

reduced pressure, then dissolved in distilled

water, and partitioned with petroleum ether

(PE, b.p 30 - 60oC) and EtOAc successively to

afford PE (15.9 g) and EtOAc (17.9 g)

fractions The PE fraction (3 g) was subjected

to FC (applied pressure ~ 0.2 mbar) eluted with

pentane-EtOAc (95 : 5) to give 6 fractions

Fraction 1 contained 33mg selinene and

-selinene 171 mg fraction 2 was

rechromato-graphed on AgNO3-impregnated silica gel

using pentane as eluent to give 34 mg

-humulene Repeated FC of 515 mg fraction 3

eluted with PE-diethyl ether (4:1) afforded

furanodiene (1) (52 mg), curzerenone (3) (43

mg) and germacrone (4) (63 mg) Fractions 4

and 5 were concentrated under reduced

pressure and further purified by solid phase

extraction using silica gel Waters cartridges to

give furanodienone (2) in the yields of 298 mg

and 424 mg, respectively Fraction 6 was

repeatedly chromatographed using

pentane-diethyl ether (7 : 3) to yield 83 mg

curcumenone (5) (GC: 83%) and 100mg crystalline zederone (6) (GC: 100%)

Furanodiene (1), furanodienone (2),

curzerenone (3) and germacrone (4) were

obtained as colorless oils For RRISil5, MS, 1H- and 13C-NMR, see reference 19

Curcumenone (3):

Colorless oil RRISil5: 1802

HR-MS: 234.1617 (C15H22O2)

1H-NMR: 0.40 (1H, ddd, J = 4.4 Hz, 7.3

Hz, 7.3 Hz, H-1), 0.64 (1H, m, H-5), 1.08 (3H,

s, 10-CH3), 1.54 - 1.62 (2H, m, 2 H-2), 1.75 (3H, s, 11-CH3), 2.06 (3H, s, 11-CH3), 2.10 (3H, s, 4-CH3), 2.44 (2H, dd, J = 7 Hz, 7 Hz, 2 H-3), 2.52 (1H) and 2.98 (1H), AB system, J =

16 Hz (2 H-9), 2.70 - 2.80 (2H, m, 2 H-6)

13C-NMR: 24.0 (d, C-1), 23.3 (t, C-2), 43.7 (t, C-3), 208.8 (s, C=O, C-4), 23.3 (d, C-5), 28.1 (t, C-6), 127.9 (s, C-7), 201.7 (s, C=O, C-8), 48.7 (t, 9), 19.9 (s, 10), 147.5 (s, 11), 23.3 (q,

C-12), 23.3 (q, C-13), 29.9 (q, C-14), 18.9 (q, C-15)

Zederone (6):

Colorless needles mp 149-150oC RRISil5:

1961

[ ]D31 +290o(c 1.14, CHCl3)

IR: max cm-1 1674, 1558, 1428, 1235, 1108,

1022, 931, 846

HR-MS: 246.1256 (C15H18O3)

GC-MS: m/z (%) 246 (35), 231 (4), 213 (5),

188 (29), 175 (100), 161 (38), 147 (22), 133 (15),

122 (25), 119 (74), 105 (24), 91 (29), 77 (23), 65 (16), 53 (12)

1H-NMR (with COSY): 1.27 (1H, ddd, J = 4

Hz, 13.0 Hz, 13.5 Hz, H-3 ), 1.32 (3H, s, 4-CH3), 1.58 (3H, s, 10-CH3), 2.09 (3H, s br, 11-CH3), 2.21 (1H, d br, J = 13.5 Hz, H-2 ), 2.28 (1H, ddd,

J = 3.5 Hz, 3.5 Hz, 13.0 Hz, H-3 ), 2.50 (1H, dddd, J = 3,5 Hz, 12.0 Hz, 13.5 Hz, 13.5 Hz,

H-2 ), 3.66 (1H) and 3.73 (1H), AB system, J = 16

Hz (2 H-9), 3.79 (1H, s, H-5), 5.46 (1H, dd br, J =

4 Hz, 12 Hz, H-1), 7.06 (1H, s br, H-12)

13C-NMR (with HMQC): 131.1 (d, C-1), 24.6 (t, C-2), 37.9 (t, C-3), 63.9 (s, C-4), 66.5

(d, C-5), 192.1 (s, C=O, C-6), 122.1 (s); 131.0

(s) (C-7; C-10), 157.0 (s, C-8), 41.8 (t, C-9),

123.2 (s, C-11), 138.0 (d, C-12), 10.2 (q, C-13),

15.7 (q, C-14), 15.1 (q, C-15)

Acknowlegements

The authors are grateful to the

International Foundation for Science (IFS,

Grant No F/2841-1) for the financial support

We thank Prof Dr Peter Weyerstahl,

Technische Universitat Berlin, Germany, for

the GC-MS and NMR spectra

REFERENCES

1 Vo Van Chi Dictionary of Vietnamese

medicinal plants, Publishing House

Medicine, Ho Chi Minh City (1997)

2 C A N Catalan, A Bardon, J A Retamar,

E G Gros, J Verghese, M T Joy Flavour

Fragrance J., 4 (1), 25 - 30 (1989)

3 M Kuroyanagi, A Ueno, K Ujiie, S Sato

Chem Pharm Bull., 35 (1), 53 - 59 (1987)

4 M Kuroyanagi, K Ujiie, A Ueno, S Sato

Tennen Yuki Kagobutsu Toronkai Koen

Yoshishu, 29, 528 - 535 (1987); see Chem

Abstr 109, 190589p (1988)

5 M Kuroyanagi, A Ueno, K Koyama and

S Natori Chem Pharm Bull., 38 (1), 55

-58 (1990)

6 F D Carter, F C Copp, B S Rao, J L

Simonsen and K S Subramaniam J Chem

Soc., 1939, 1504 - 1509

7 A Gopalam, M J Ratnambal Indian

Perfum., 31 (3), 245 - 248 (1987)

8 Y T Guo, X K Chu, Y R Chen, X Y

Wu, G Chen, X R Lu Yao Hsueh Hsueh

Pao, 15 (4), 251 - 252 (1980); see Chem

Abstr 93, 235081m (1980)

9 J T Rao, S S Nigam Flavour Ind., 5

(9-10), 234 - 236 (1974)

10 B S Rao, V P Shintre and J L Simonsen

J Indian Inst Sci., 9A, 140 - 144 (1926); see Chem Abstr 21, 7988(1927)

11 B S Rao and J L Simonsen J Chem Soc.,

2496 - 2505 (1939)

12 J H Zwaving, R Bos Flavour Fragrance

J., 7 (1), 19 - 22 (1992)

13 Phan Tong Son, Van Ngoc Huong, Nguyen Van Dau, Luong Si Binh, M A Posthumus

J of Chemistry (Vietnam), 27 (3), 18 - 19

(1989)

14 Luong Si Binh Ph.D Thesis, Hanoi University (1987)

15 Phan Minh Giang, Van Ngoc Huong, Phan

Tong Son J of Chemistry (Vietnam), 37

(1), 57-59 (1999)

16 H Hikino, K Agatsuma, T Takemoto

Tetrahedron Lett., 8, 931 - 933 (1968)

17 H Hikino, C Konno, K Agatsuma, and T Takemoto, I Horibe, K Tori, M Ueyma, and

Ken'ichi Takeda J Chem Soc., Perkin Trans.

I, 1975 (5), 478 - 484

18 H Hikino, K Agatsuma, T Takemoto

Tetrahedron Lett., 24, 2855 - 2858 (1968)

19 Phan Minh Giang, Van Ngoc Huong, Phan

Tong Son J of Chemistry (Vietnam), 36

(3), 67 - 72 (1998)

20 Y Shiobara, Y Asakawa, M Kodama, K Yasuda and T Takemoto Phytochemistry,

24 (11), 2629 - 2633 (1985)

21 M Ohshiro, M Kuroyanagi and A Ueno,

Phytochemistry, 29(7), 2201 - 2205 (1990)

22 H Hikino, H Takahashi, Y Sakurai, T Takemoto, N S Bhacca Chem Pharm

Bull., 14 (5), 850 - 851 (1966)

23 H Hikino, K Tori, I Horibe, and K Kuriyama J Chem Soc (C), 1971, 688

-691

24 H Shibuya, Y Hamamoto, Y Cai, and I

Kitagawa Chem Pharm Bull., 35 (2),

924-927 (1987)

100