A new secoiridoid glycoside and a new sesquiterpenoid glycoside from Valeriana jatamansi with neuroprotective activity

Article abstract of DOI:10.1016/j.phytol.2016.07.020

A new secoiridoid glycoside, isopatrinioside (1) and a new sesquiterpenoid glycoside, valeriananoid F (2), together with nine known compounds, were isolated from the roots of Valeriana jatamansi. Their structures were elucidated on the basis of spectroscopic analysis. Compound 1 was an unusual monocyclic iridoid glycoside ring-opened between C-1 and C-2 produced by the cleavage of the pyran ring. Of the eleven isolates, compounds 1 and 4 exhibited moderate neuroprotective effects against CoCl₂-induced neuronal cell death in PC12 cells.

Full text of DOI:10.1016/j.phytol.2016.07.020

Phytochemistry Letters 17 (2016) 177–180
Contents lists available at ScienceDirect
Phytochemistry Letters
journal homepage: www.elsevier.com/locate/phytol
A new secoiridoid glycoside and a new sesquiterpenoid glycoside from
Valeriana jatamansi with neuroprotective activity
Yu-Zhu Tana,b, Yan Yonga,b, Yan-Hong Donga,b, Ru-Jing Wanga,b, Hong-Xiang Lib,
a,b,**
Hai Zhangb, Da-Le Guoa,b, Shi-Jin Zhangb, Xiao-Ping Dongb, , Xiao-Fang Xie
*
a
State Key Laboratory Breeding Base of Systematic Research Development and Utilization of Chinese Medicine Resources, Sichuan Province and Ministry of
Science and Technology, Chengdu 611137, China
b
Pharmacy College, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China
A R T I C L E I N F O
A B S T R A C T
Article history:
A new secoiridoid glycoside, isopatrinioside (1) and a new sesquiterpenoid glycoside, valeriananoid F (2),
together with nine known compounds, were isolated from the roots of Valeriana jatamansi. Their
structures were elucidated on the basis of spectroscopic analysis. Compound 1 was an unusual
monocyclic iridoid glycoside ring-opened between C-1 and C-2 produced by the cleavage of the pyran
ring. Of the eleven isolates, compounds 1 and 4 exhibited moderate neuroprotective effects against
CoCl2-induced neuronal cell death in PC12 cells.
Received 4 May 2016
Received in revised form 20 June 2016
Accepted 6 July 2016
Available online xxx
Keywords:
Valeriana jatamansi
Secoiridoid glycoside
Sesquiterpenoid glycoside
Neuroprotective activity
ã 2016 Phytochemical Society of Europe. Published by Elsevier Ltd. All rights reserved.
1. Introduction
nine known compounds. Isopatrinioside (1) is a monocyclic iridoid
glycoside with pyran ring-opened between C-1 and C-2. To the best
Valeriana jatamansi Jones belongs to the family Caprifoliaceae,
an annual herb distributed widely in the southwest of China and
India, which is well known as traditional Chinese medicine with
tranquilizing hypnotic, nervous disorders, epilepsy, insanity, snake
poisoning and skin diseases (Ming et al.,1997; Mathela et al., 2005;
Fernandez et al., 2004). Its root, as an important substitute of
European V. officinalis, has been used to treat nervous disorders
(Mathela et al., 2005; Fernandez et al., 2004). Previous chemical
investigation of V. jatamansi revealed the presence of iriodoids,
sesquiterpenoids, essential oil, flavone glycosides and lignans
(Ming et al., 1997; Tang et al., 2003; Verma et al., 2011; Lin et al.,
2010).
of our knowledge, the report about this type monocyclic iridoid
from natural sources is still very rare (Wysokinska and Skrzypek,
1992; Dinda et al., 2007, 2009, 2011). Isopatrinioside(1) is so far
another positional isomer of patrinioside isolated from valeriana
(Kouno et al., 1995). Valeriananoid F(2) is one of the valeriananoid
analogues which have been isolated from V. jatamansi (Ming et al.,
1997; Dong et al., 2015). In this paper, we describe the isolation,
structural elucidation of two new compounds and neuroprotective
effects of the eleven compounds.
2. Results and discussion
In the course of our continual search for bioactive natural
products on nervous systems from Valeriana genus, eleven
compounds were isolated from the roots of Valeriana jatamansi
including a new secoiridoid glycoside, isopatrinioside (1) and a
new sesquiterpenoid glycoside, valeriananoid F (2), together with
Compound 1 was obtained as an amorphous powder. The IR
spectrum showed typical absorption bands for hydroxy
(3400 cmꢀ1), ester carbonyl (1713 cmꢀ1), and glycosyl (1077,
1047 cmꢀ1). The molecular formula was determined to be
C
21H36O10 on the basis of the HRESIMS ([M + Na]+ at m/z
471.2204, calcd. for 471.2206) with 4 degrees of unsaturation.
The 1H NMR (Table 1), DEPT combinated with HSQC spectra of 1
showed a doublet methyl signals at dC 22.8 and dH 0.96, methylene
carbon signals (dC 45.3), a methine (dC 26.9), and a carbonyl carbon
signal at dC 174.4, indicating an isovaleryloxyl ester moiety[dH 0.96
(6H, d, J = 6.6 Hz), 2.05 (1H, m), 2.16 (2H, m); dC 174.4, 45.3, 26.9,
22.8], the presence of which was confirmed by correlations in the
* Corresponding author at: Pharmacy College, Chengdu University of Traditional
Chinese Medicine, Chengdu 611137, China.
** Corresponding author at: State Key Laboratory Breeding Base of Systematic
Research Development and Utilization of Chinese Medicine Resources, Sichuan
Province and Ministry of Science and Technology, Chengdu 611137, China.
E-mail addresses: dongxiaoping11@126.com (X.-P. Dong), xxf14544@163.com
(X.-F. Xie).
http://dx.doi.org/10.1016/j.phytol.2016.07.020
1874-3900/ã 2016 Phytochemical Society of Europe. Published by Elsevier Ltd. All rights reserved.

178
Y.-Z. Tan et al. / Phytochemistry Letters 17 (2016) 177–180
that 1 was a monocyclic iridoid ring-opened. 1H and 13C NMR
spectroscopic data showed the compound gross structure was
closely similar to jatamanin J except for the absence of one hexose
and isovaleryl moiety (Lin et al., 2010). Detailed analysis of 2D NMR
data verified the location of the substitutents. The HMBC
correlations (Fig. 1) between the Glc anomeric proton at dH 4.13
(1H, d, J = 7.8 Hz) and C-1 (dC 68.0) indicated that the glucose
moiety was linked to C-1 (dC 68.0) of the aglycone. In the HMBC
spectrum (Fig.1), the position of the isovaleryloxyl ester group was
determined as C-8 (dC 93.1) on the basis of weak correlation of H-
10 (dH 1.6) and the ester carbonyl of the isovaleroxyl moiety. The
stereo-structure was examined by a NOESYexperiment. The NOESY
correlations of H-9 (dH 2.57) with H-5 (dH 2.97) and H-10 (dH 1.6),
Table 1
1H and 13C NMR Data (600 and 150 MHz, resp., MeOD-d4) of 1 and 2 with J values
(Hz) in parentheses.
NO.
1
2
dH
dC
dH
dC
1a
1b
2
3a
3b
4
5
6a
6b
7a
7b
8
9
10
11a
11b
12
4.02 dd (9.6,4.0)
3.50 dd (9.6,8.6)
68.0
–
2.05 dd (14.0,7.6)
1.60 dd (14.0,6.7)
37.8
–
–
–
3.96 ddd (7.6,6.7,1.2)
1.48 t (5.9)
80.4
46.0
–
5.28 d (1.1)
4.98 bs
110.2
–
–
–
150.7
40.2
37.1
–
–
40.0
76.6
32.8
–
2.97 br q (9.1)
2.12 ddd (14.5,6.8,1.9)
1.72 ddd (14.5,8.6,3.6)
–
1.73 dd (14.3,5.2)
1.54 m
4.49 dd (6.8,3.6)
–
76.7
–
1.52 m
1.38 m
29.6
–
H-7 (dH 4.49) with H-6
1.72) showed that the H-5 (dH 2.97) and H-9 (dH 2.57) possess a cis-
configuration and indicated -OH, -isovaleroxyl groups.
a (dH 2.12), H-5 (dH 2.97) with H-6b (dH
–
93.1
51.4
19.2
66.7
–
1.96 m
1.36 m
29.0
44.3
41.5
25.0
–
7b
8a
2.57 td (8.6,4.0)
1.6 s
4.10 br d (14.5)
4.00 br d (14.5)
–
–
Comparisions of NOESY NMR data for 1 with those from jatamanin
J allowed the relative configurations at C-5 (dC 40.2), C-7 (dC 76.7),
C-8 (dC 93.1), and C-9 (dC 51.4) to be identically assigned (Lin et al.,
2010). The above data strongly suggested that 1 is a position isomer
of patrinioside with an isovaleroxyl group at C-8 (dC 93.1) and
sugar moiety at C-1 (dC 68.0). Thus, the structure of 1 was
1.55 m
1.32 m
0.81 d (6.7)
1.09 s
1.17 s
0.89 s
4.33 d (7.8)
3.15 dd (8.8,7.9)
3.35 br t (8.8)
3.29 m
3.24 m
3.86 dd (11.9,2.3)
–
19.3
25.7
28.9
21.0
103.3
75.3
77.8
71.8
76.6
62.9
–
13
14
15
–
–
–
–
–
–
Glc-1
Glc-2
Glc-3
Glc-4
Glc-5
Glc-6a
Glc-6b
10
4.13 d (7.8)
3.13 dd (8.8,7.9)
3.33 br t (8.8)
3.27 br t (9.5)
3.23 m
3.85 dd (11.9,2.3)
3.66 dd (11.9,5.6)
–
104.6
75.2
78.1
71.6
77.8
62.7
–
established as 8-O-isovaleroxyl-1-O-b-D-glucopyranosyl jatama-
nin J and named isopatrinioside.
Compound 2 was obtained as a white, amorphous powder. 2
had the molecular formula C21H36O7, which was determined by
HRESIMS ([M + Na]+ at m/z 423.2352, calcd. for 423.2359),
requiring 4 unsaturation. The IR spectrum of 2 displayed the
3.67 dd (11.9,5.6)
174.4
45.3
26.9
22.8
22.8
–
–
–
–
–
–
presence of the hydroxyl (3414 cmꢀ1
)
and glycosyl
20
2.16 m
2.05 m
0.96 d (6.6)
0.96 d (6.6)
–
(1074,1045 cmꢀ1). The 1H NMR spectrum (Table 1) of 2 exhibited
four methyls [dH 0.81 (3H, d, J = 6.7 Hz),1.17 (3H, s),1.09 (3H, s), 0.89
30
–
40
–
50
–
(3H, s)], and anomeric proton signal of b-glycosyl group at dH 4.33
(1H, d, J = 7.8 Hz), which gave correlation in the HMQC spectrum at
dC 103.3. The 13C NMR (Table 1) and DEPT spectroscopic data, in
combination with HMQC spectrum, revealed the existence of one
tricyclic sesquiterpenoid skeleton, including four methyls [dH 1.17
(1H, s),1.09 (1H, s), 0.89 (1H, s), 0.81 (1H, d, J = 6.7 Hz); dC 28.9, 25.7,
21.0, 19.3]; four methylenes [dH 2.05 (1H, dd, J = 14.0, 7.6 Hz), 1.60
(1H, dd, J = 14.0, 6.7 Hz), 1.73 (1H, dd, J = 14.3, 5.2 Hz), 1.54 (1H,m),
1.52 (1H,m), 1.38 (1H,m), 1.55 (1H, m), 1.32 (1H, m); dC 37.8, 32.8,
29.6, 25.0], four methines [dH 3.96 (1H, ddd, J = 7.6, 6.7, 1.2 Hz), 1.48
(1H, t, J = 5.9 Hz), 1.36 (1H,m), 1.96 (1H,m); dC 80.4, 46.0, 44.3, 29.0]
as well as three quaternary carbons [dC 40.0, 41.5, 76.6], which
according to remaining three unsaturated degree. A careful
comparison of the NMR data of 2 with those of the known
compound valeriananoid B (Mathela et al., 2005) indicated that
they were structural analogues. The only difference was absence
1H-1H COSY and HSQC spectrum. In addition, the presence of a
-glycosyl moiety was established by an anomeric proton [dH 4.13
b
(1H, d, J = 7.8 Hz)] and partially overlapped signals attributed to
oxymethylene and oxymethine protons between dH 3.13 and 3.85.
Acid hydrolysis afforded D-glucose as the sole sugar, identified by
20
TLC comparison and the positive optical rotation ½aꢁD þ 47:0ꢂ (Gan
et al., 2008; Hudson and Dale, 1917). Five of the remaining 10
carbon signals were ascribable to a methyl (dC 19.2), exo-
methylene (dC 150.7 and 110.2), and two oxy-methylene (dC
66.7 and 68.0).
The connectivities of the proton coupling sequence for the C-1-
C-9-C-5-C-6-C-7 fragment in an iridoid skeleton were observed in
the 1H-1H COSY spectrum (Fig. 1). The absence of HMBC
correlations between H-3 (dH 5.28 and 4.98) and C-1 (dC 68.0)
and/or between H-1 (dH 4.02 and 3.50) and C-3 (dC 110.2) showed
the
b-D-glucose moiety in valeriananoid B. The position of b-D-
glucose at C-2 (dC 80.4) was confirmed based on the HMBC
15
OH
11
H
OH
H
H
H
8
O
1'
H
9
4
12
5
O
H
O
3
1
HO
HO
OH
1''
HO
O
O
OH
OH
HO
HO
1
10
OH
13
O
14
2
1
1H,1H COSY
HMBC(H
C)
NOESY
Fig. 1. Key 1H, 1H COSY, HMBC and NOSEY correlations of compound 1 and 2.

Y.-Z. Tan et al. / Phytochemistry Letters 17 (2016) 177–180
179
correlation (Fig. 1) from anomeric proton dH 4.33 (1H, d, J = 7.8 Hz)
to C-2 (dC 80.4). A detailed analysis of HMQC, HMBC and 1H-1H
COSY spectrum (Fig.1) supported the assignment of all protons and
carbon signals. The relative configuration of 2 was established by
Preparative TLC was conducted with glass plates precoated silica
gel GF254 (Yantai). TLC was carried out with GF254 plates (Qingdao
Marine Chemical Factory). Spots were visualized by spraying with
10% H2SO4 in 95% EtOH followed by heating. Column chromatog-
raphy (CC) was performed with silica gel and Sephadex LH-20. All
the solvent used were of analytical grade.
the NOESY experiment. The NOESY correlations of H-1
with H-3 (dH 1.48), H-15 (dH 0.89) with H-3 (dH 1.48), H-8 (dH 1.96)
with H-15 (dH 0.89) and H-14 (dH 1.17) with H-1 dH 2.05). The
a (dH 2.05)
a
(
relative configuration of 2 was the same as that of valeriananoid B
and detailed comparison of the 1H NMR coupling constants of 2
with valeriananoid B (Mathela et al., 2005). Consequently, 2 was
defined and named as valeriananoid F.
3.2. Plant material
The root of Valeriana jatamansi Jones was collected in June of
2014 from the medicinal herbs market in Chengdu, Sichuan
Province, China. The medical material identity was verified by Prof.
Min Li (Chengdu University of TCM, Sichuan, China). A voucher
specimen (ZZX-1407) was deposited at the School of Pharmacy,
Chengdu University of TCM, Chengdu, China.
In addition to the two new compounds 1 and 2, the known
iridoids and sesquiterpenoid were identified by comparison of
spectroscopic data with those reported in the literature as 8-
methoxy-3-methoxy-10-methylene-2,9-dioxatricyclo[4.3.1.03,7
]
decan-4-ol (3) (Inouye et al., 1974), vibutinal (4) (Liu et al., 2010),
baldrinal (5) (Chen et al., 2005),11-ethoxyviburtinal (6) (Boros and
Stermitz, 1991), jatamanvaltrate E (7) (Lin et al., 2009), jataman-
valtrate A (8) (Lin et al., 2009), jatamanvaltrate F (9) (Lin et al.,
2009), valeriotetrate C (10) (Lin et al., 2010), chlorovaltrate A (11)
(Lin et al., 2013).
3.3. Extraction, isolation and characterization of compounds
The air-dried roots of Valeriana jatamansi Jones (5 kg), was
powdered and exhaustively extracted using by percolating with
75% EtOH (3 ꢃ 30 L). The combined extract was concentrated under
reduced pressure to dryness. The residue was suspended in water
(2 L) and then the successively partitioned with Petroleum Ether
(3 ꢃ 2 L), EtOAc (3 ꢃ 2 L)and n-BuOH (3 ꢃ 2 L). The EtOAc extract
(170 g) was subjected to CC over silica gel and eluted with a
gradient of CHCl3-MeOH [95:5; 90:10; 85:15; 80:20; 75:25; 70:30;
60:40; 50:50; MeOH] to afford nine fractions A-I. Fraction D (3 g)
and E (4 g) were separated by MHPLC (3.6 cm ꢃ 46 cm, ODS, 10 mL/
min, 6 h), eluted with 40%, 50%, 60%, 70%, 80%, 90%, 100% MeOH-
H2O to afford about 7 subfractions respectively. The fraction D4
(847 mg) was chromatographed over Sephadex LH-20 eluted with
MeOH as mobile phase to give three subfractions. Successive
separation of the D4-2 (85 mg) by preparative TLC with CHCl3-
MeOH (4:1, v/v) to yield 1 (11.5 mg). The subfraction D4-3 (127 mg)
was separated by preparative TLC with Pe-EtOAc (15:1, v/v) to yield
4 (15.4 mg) and 5 (10.7 mg). Fraction D5 (215 mg)was further
purified by CC over Sephadex LH-20 eluted with MeOH-H2O (9:1, v/
v) as mobile phase to give five subfractions. The subfraction D5-2
(58 mg) was purified by preparative TLC with petroleum ether-
EtOAc (20:1, v/v) to 7 (11.2 mg). The Fraction D5-3 (48 mg)was
purified by preparative HPLC using 75% MeOH-H2O (3 mL/min) as
mobile phase to afford 2 (8.8 mg) and 6 (6.7 mg). At last, Fraction
D5-4 (45 mg) was purified by preparative HPLC using 70% MeOH-
H2O (3 mL/min) as mobile phase to afford 10 (10.5 mg), 11(8.5 mg).
Fraction E3 (86 mg) was purified by preparative HPLC using 75%
MeOH-H2O (3 mL/min) as mobile phase to yield 3 (7.5 mg) and 8
(10.5 mg). The fraction E4 (118 mg) was chromatographed over
Sephadex LH-20 eluted with MeOH as mobile phase to give four
subfractions. The subfraction E4-3 (50 mg) was purified by
preparative TLC with petroleum ether-EtOAc (10:1,v/v) to yield 9
(12.5 mg).
The neuroprotective effects of the compounds against CoCl2-
induced neuronal cell death in PC12 cells were investigated by an
established CCK-8 assay. At the concentrations of 0.1
mM, 1 mM,
10 M, compound 1 and 4 showed potent neuroprotective effects.
m
Compound 1 increased the cell viability from 56.1% to 60.9%, 63.4%
and 65.4%, respectively. For compound 4, the weak neuroprotective
activities were exhibited with the viability by 60.4% only with the
concentrations of 10
inactive even at the high concentrations of 10
m
M. However, the other compounds were
M. CCK-8 assay
m
indicated that all of the compounds had no significant cytotoxicity
to the PC12 cells at their effective concentration required for
neuroprotective effects (data not shown).
In the course of our survey on pharmacologically active
substances in Chinese herb medcine, much attention has been
paid to the occurrence of compounds with neuroprotective effects,
since these compounds are expected to be potentially useful for the
treatment and prevention of Parkinson's disease (PD). During our
search for new types of natural products possessing neuro-
protective activities, we investigated the chemical constituents of
the roots of V. jatamansi, of which the ethylacetate fractions
showed moderate neuroprotective effects. By bioactivity-guided
isolation, we isolated and identified a new secoiridoid glycoside (1)
and a new sesquiterpenoid glycoside (2) and nine known (3–11)
compounds successfully from the roots of V. jatamansi. This study
discovered secoiridoid and sesquiterpenoid glycoside from Valeri-
ana and first reported the activity of these compounds against
PC12 cell damage. It can be seen that secoiridoid glycoside was
bioactive component in V. jatamansi, which has been used as
neurological disorder drug for thousands of years in East Asia. Of
course, the neuroprotective effect of the secoiridoid aglycone is
also worth of our concern in the further research.
3.3.1. Isopatrinioside (1)
20
Amorphous powder; ½aꢁD ꢀ 15:6ꢂ (c = 0.02, MeOH); UV (MeOH)
3. Experimental
lmax (log e
): 206 (3.87), 220 (3.77); IR (KBr) nmax cmꢀ1: 3400, 2958,
3.1. General experimental procedures
1713, 1605, 1077, 1047; HRESIMS m/z: 471.2204 [M+Na]+ (calcd for
C21 H36 O10Na 471.2206); 1H and 13C NMR data see Table 1.
NMR measurements were performed on a Bruker-AVII-600
spectrometer. HRESIMS were obtained using Waters Synapt
G2HDMS. IR spectra were measured by PerkinElmer one FT-IR
spectrometer (KBr). UV spectra were obtained on a Shimadzu UV-
260 spectrophotometer. Optical rotations were measured with
Anton Paar MCP 200. Preparative HPLC was performed on
Shimadazu LC-10AT instrument with an SPD-10AVP detector
3.3.2. Valeriananoid F(2)
20
Amorphous powder; ½aꢁD ꢀ 18:6ꢂ (c = 0.05, MeOH); UV (MeOH)
lmax (loge :
): 209 (3.27), 220 (3.14), 228 (3.16); IR (KBr) nmax cmꢀ1
3414, 2926, 2870, 1574, 1416, 1074, 1045; HRESIMS m/z: 423.2352
[M+Na]+ (calcd for C21H36O7Na 423.2359); 1H and 13C NMR data
see Table 1.
and
a
YMC-Pack ODS-A column (250 mm ꢃ 10 mm,
5 mm).

180
Y.-Z. Tan et al. / Phytochemistry Letters 17 (2016) 177–180
3.4. Acid hydrolysis
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20
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Financial support from the project for National Science
Foundation for Fostering Talents in Basic Research of the National
Natural Science Foundation of China fund (grant No. J13100340-
11), the key fund project for Education Department of Sichuan
(grant No.15ZA0093), the youth science and technology innovative
research team fund project of Sichuan (grant No. 2016TD0006) and
the project for administration of Traditional Chinese Medicine
(grant No. 2016Q049).
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Appendix A. Supplementary data
Supplementary data associated with this article can be found, in
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Phytochemistry 31, 1235–1237.
the
online
version,
at
http://dx.doi.org/10.1016/j.
phytol.2016.07.020.