2018

NPC Natural Product Communications Vol. 13

No. 2
Chemical Constituents of Vitex trifolia Leaves 129 - 130
Ninh Khac Bana, Nguyen Thi Kim Thoab, Tran My Linha, Vu Huong Gianga, Do Thi Tranga, Nguyen Xuan
Nhiema, Bui Huu Taia, Tran Hong Quanga, Pham Hai Yena, Chau Van Minha and Phan Van Kiema,*
a
Institute of Marine Biochemistry, Vietnam Academy of Science and Technology (VAST), 18 Hoang Quoc Viet,
Cau Giay, Hanoi, Vietnam
b
Faculty of Basic Sciences, Hanoi University of Mining and Geology, 18 Pho Vien, Bac Tu Liem, Hanoi, Vietnam

[email protected]

Received: December 1st, 2017; Accepted: January 12th, 2018

Using various chromatographic methods, one new lanostane triterpene, 3α-hydroxylanosta-8,24E-dien-26-oic acid (1), one new lignan, matairesinol 4′-O-β-D-
glucopyranoside (2) along with five known compounds, ecdysone (3), 20-hydroxyecdysone (4), 20-hydroxyecdysone 2,3-monoacetonide (5), turkesterone (6),
and polypodine B (7) were isolated from the leaves of Vitex trifolia L. Their structures were elucidated by 1D-, 2D-NMR spectroscopic analysis, HR-ESI-MS,
CD, and by comparing with the NMR data reported in the literature.

Keywords: Vitex trifolia, Lamiaceae, Lanostane, 3α-Hydroxylanosta-8,24E-dien-26-oic acid, Matairesinol 4′-O-β-D-glucopyranoside.

Vitex is the genus of shrubs and trees and mainly distributed in

tropics and subtropics. Since ancient times, civilization used Vitex
plants for treating many health problems such as malaria, herpes,
itches, dermatitis or controlling menstruation [1]. Phytochemical
study of Vitex trifolia has led to the isolation of diterpenoids,
triterpenoids, flavonoids [2a-d], and many others. Some of them
exhibited the inhibitory anti-cancer and anti-inflammatory activities
[3]. Herein, we report the isolation, structure elucidation of two new
and five known compounds from V. trifolia.

Compound 1 was obtained as a white amorphous powder. Its Figure 1: Chemical structures, key HMBC, and COSY of 1 and 2
molecular formula was determined as C30H48O3 by HR-ESI-MS at
m/z 455.3540 [M‒H]‒ (Calcd. for [C30H47O3]‒, 455.3531). The 1H- Compound 2 was obtained as a white amorphous powder. Its
NMR spectrum of 1 showed the proton signals of 7 methyl groups at molecular formula was determined as C26H32O11 on the basic of
0.69 (s), 0.87 (s), 0.88 (s), 0.94 (d, J = 6.5 Hz), 0.97 (s), 0.99 (s) and HR-ESI-MS ion at m/z 521.2009 [M+H]+ (Calcd. for [C26H33O11]+,
1.84 (s), one olefinic proton at δH 6.89 (1H, t, J = 7.5 Hz), and one 521.2017). The 1H-NMR spectrum of 2 (in CD3OD) showed the
oxymethine proton at δH 3.43 (br d, J = 3.0 Hz). The 13C-NMR and signals for two pairs of ABX aromatic protons at δH 6.49 (dd, J =
DEPT spectra of 1 exhibited the signals of 30 carbons, including one 1.6, 8.0 Hz), 6.55 (d, J = 1.6 Hz), and 6.66 (d, J = 8.0 Hz); 6.64 (dd,
carbonyl carbon, seven quaternary carbons, five methines, ten J = 1.6, 8.0 Hz), 6.73 (d, J = 1.6 Hz), and 7.04 (d, J = 8.0 Hz), two
methylenes, and seven methyl carbons. Analysis the NMR data of methoxy groups at δH 3.75 (s) and 3.79 (s), one anomeric proton at
compound 1 suggested that structure of 1 was lanostane skeleton and δH 4.84 (d, J = 8.0 Hz) as listed in Table 1. The 13C-NMR and
similar to the astraodoric acid D except for the disappearance of DEPT spectra of 2 revealed 26 carbon signals, of which, 18 were
acetoxy group at C-22 [4a]. The HMBC correlations from H-28 (δH assigned to a lignan moiety, 2 carbons belonged to two methoxy
0.88)/H-29 (δH 0.97) to C-3 (δC 76.1)/C-4 (δC 37.6)/C-5 (δC 44.3) groups, and 6 carbons contributed to a sugar moiety. The 1H- and
13
indicated the presence of the hydroxyl group at C-3. The C-NMR data of 2 were very similar to those of matairesinol 4-O-
configuration of the hydroxyl group is defined as α (axial) because of β-D-glucopyranoside [4d] except for the change position of
the small coupling constant between H-2 and H-3 (JH-2-H-3 = 3.0 Hz). glucopyranosyl moiety from C-4 to C-4′. Butanolide ring was
The HMBC correlations between H-19 (δH 0.99) and C-1 (δC 30.1)/C- confirmed by HMBC correlation between H-9 (δH 3.91 and 4.16)
5 (δC 44.3)/C-9 (δC 134.7)/C-10 (δC 36.9); between H-30 (δH 0.87) and C-9′ (δC 181.5) as well as COSY correlations of H-8′ (δH
and C-8 (δC 134.1)/C-13 (δC 44.6)/C-14 (δC 49.9)/C-15 (δC 30.8) 2.66)/H-8 (δH 2.47)/H-9 (δH 3.91 and 4.16). The HMBC correlations
confirmed the presence of the double bond at C-8/C-9. The HMBC between H-7 (δH 2.52) and C-1 (δC 131.3)/C-2 (δC 113.3)/C-6 (δC
correlations between H-27 (δH 1.84) and C-24 (δC 145.7)/C-25 (δC 122.2)/C-8 (δC 42.6)/C-9 (δC 72.9)/C-8′ (δC 47.6); between methoxy
126.6)/C-26 (δC 172.7) indicated the double bond at C-24/C-25 and protons (δH 3.75) and C-3 (δC 149.0); between H-7′ (δH 2.85) and C-
the carboxylic group at C-25. Moreover, the chemical shift of C-27 1′ (δC 134.2)/C-2′ (δC 114.8)/C-6′ (δC 123.0)/C-8′ (δC 47.6)/C-9′ (δC
(δC 12.0) proved E configuration of the double bond C-24/C-25, 181.5)/C-8 (δC 42.6); and between methoxy protons (δH 3.79) and
which based on the comparison of 13C-NMR chemical shift (δC 12.0, C-3′ (δC 150.6) suggested the positions of two 3-methoxy-4-
C-27) of 7-oxo-ganoderic acid Z (E configuration) [4b] and 13C-NMR hydroxyphenyl groups at C-7 and C-7′. The coupling constant (JH-
13
chemical shift (δC 22.8, C-27) of schiglauzic acid (Z configuration) 1″/H-2″ = 8.0 Hz) of the anomeric proton of sugar moiety and its C-
[4c]. From the above evidence, the new structure of 1 was NMR chemical shifts: C-1′′ (δC 102.9), C-2′′ (δC 74.9), C-3′′ (δC
elucidated to be 3α-hydroxylanosta-8,24E-dien-26-oic acid. 77.8), C-4′′ (δC 71.3), C-5′′ (δC 78.1), and C-6′′ (δC 62.5) suggested
130 Natural Product Communications Vol. 13 (2) 2018 Ban et al.

Table 1: The 1H- and 13C-NMR data for compounds 1 and 2. The known compounds were identified as ecdysone (3) [5a], 20-
C 1 C 2 hydroxyecdysone (4) [5b], 20-hydroxyecdysone 2,3-monoacetonide
δCa,c δHa,d (mult., J, Hz) δCb,e δHb,f (mult., J, Hz) (5) [5c], turkesterone (6) [5d], and polypodine B (7), [5e] by
1 30.1 1.27 (m)/1.48 (m) 1 131.3 -
2 25.7 1.63 (m)/1.95 (m) 2 113.3 6.55 (d, 1.6)
comparison their NMR with the reported values in the literature.
3 76.1 3.43 (br d, 3.0) 3 149.0 -
4 37.6 - 4 146.2 - Experimental
5 44.3 1.53 (m) 5 116.2 6.66 (d, 8.0)
6 18.2 1.53 (m)/1.60 (m) 6 122.2 6.49 (dd, 1.6, 8.0)
General: See supporting information.
7 25.9 2.05 (m) 7 38.9 2.52 (m)
8 134.1 - 8 42.6 2.47 (m) Plant materials: The leaves of V. trifolia L. were collected in Bach
9 134.7 - 9 72.9 3.91 (dd, 8.0, 8.8, α) Ma National Park, Thua Thien Hue, Vietnam in September, 2015,
10 36.9 - 4.16 (dd, 7.2, 8.8, β) and authenticated by one of the authors, Prof. N. K. Ban. A voucher
11 21.0 2.04 (m) 3-OMe 56.4 3.75 (s) specimen was deposited at Institute of Marine Biochemistry, VAST.
12 31.0 1.60 (m)/1.72 (m) 1′ 134.2 -
13 44.6 - 2′ 114.8 6.73 (d, 1.6)
14 49.9 - 3′ 150.6 -
Extraction and isolation: The dried leaves of V. trifolia were
15 30.8 1.20 (m)/1.61 (m) 4′ 146.8 - extracted with hot MeOH three times using sonicator to yield
16 28.2 1.32 (m)/1.93 (m) 5′ 117.8 7.04 (d, 8.0) extract after evaporation of the solvent. This extract was suspended
17 50.3 1.52 (m) 6′ 123.0 6.64 (dd, 1.6, 8.0) in H2O and successively partitioned with CH2Cl2, EtOAc to obtain
18 15.8 0.69 (s) 7′ 35.3 2.85 (dd, 6.8, 12.8) the CH2Cl2, EtOAc, and H2O extracts, respectively after removal of
19 19.0 0.99 (s) 9′ 181.5 -
20 36.4 1.44 (m) 8′ 47.6 2.66 (m)
the solvents in vacuo. Using various chromatographic methods,
21 18.5 0.94 (d, 6.5) 3′-OMe 56.7 3.79 (s) seven compounds 1-7 were isolated from above extracts. For detail,
22 34.8 1.18 (m)/1.58 (m) 4′-O-glc see supporting information.
23 26.1 2.10 (m)/2.27 (m) 1′′ 102.9 4.84 (d, 8.0)
24 145.7 6.89 (t, 7.5) 2′′ 74.9 3.46 (dd, 8.0, 9.2) Acid hydrolysis: See supporting information.
25 126.6 - 3′′ 77.8 3.45 (m)
26 172.7 - 4′′ 71.3 3.38 (m) 3α-Hydroxylanosta-8,24E-dien-26-oic acid (1)
27 12.0 1.84 (s) 5′′ 78.1 3.38 (m)
28 24.3 0.88 (s) 6′′ 62.5 3.67 (dd, 4.0, 10.8)
White amorphous powder.
29 28.0 0.97 (s) 3.85 (br d, 10.8) [α]D25: +50.0 (c 0.1, MeOH).
1
30 22.1 0.87 (s) H- and 13C-NMR (CD3OD): Table 1.
a)
Recorded in CDCl3; b)CD3OD; c)125 MHz; d)
500 MHz; e)100 MHz; f)400 MHz HR-ESI-MS m/z: 455.3540 [M-H]‒ (Calcd. for [C30H47O3]‒,
455.3531).
the presence of β-D-glucopyranosyl moiety. This was further
confirmed by acid hydrolysis of 2 (identified as TMS derivative). In Matairesinol 4′-O-β-D-glucopyranoside (2)
addition, the HMBC correlation between glc H-1″ (δH 4.84) and C- White amorphous powder.
4′ (δC 146.8) determined the glucose moiety at C-4′ of the aglycone. [α]D25: –18.0 (c 0.1, MeOH).
The absolute configuration of aglycone was determined by the CD (c=1.0×10‒3, MeOH): [θ]25 (rel, nm) ‒1.00 (226), ‒0.21 (275).
NOESY and CD spectra. The NOE correlations between H-8′ (δH 1
H- and 13C-NMR (CD3OD): Table 1.
2.66) and Hα-9 (δH 3.91); H-8 (δH 2.48) and Hβ-9 (δH 4.16)/H-7 (δH HR-ESI-MS m/z: 521.2009 [M+H]+ (Calcd. for [C26H33O11]+,
2.52); H-7 (δH 2.52) and Hα-9 (δH 3.91), suggested the 521.2017).
configurations of H-8 and H-8′ to be trans. The two negative Cotton
effects at 226 and 275 nm in the CD spectrum indicated a (8R,8′R)- Acknowledgment - This research is funded by Vietnam National
configurations in matairesinol [4d]. Consequently, the new Foundation for Science and Technology Development
compound 2 was determined to be matairesinol 4′-O-β-D- (NAFOSTED) under grant number 104.01-2014.02.
glucopyranoside.

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