Two new dammarane-type triterpenoids from the steamed roots of Panax notoginseng

Article abstract of DOI:10.1080/10286020.2020.1825391

Two new dammarane-type triterpenoids, notoginsenoside SY3 (1) and notoginsenoside SY4 (2), were isolated from the steamed roots of Panax notoginseng. Their structures were determined to be 3β, 12β, 20(S)-trihydroxy-27-anordammar-23(24)(E)-ene-3-O-β-D-gluc

Full text of DOI:10.1080/10286020.2020.1825391

Journal of Asian Natural Products Research
ISSN: (Print) (Online) Journal homepage: https://www.tandfonline.com/loi/ganp20
Two new dammarane-type triterpenoids from the
steamed roots of Panax notoginseng
Chong-Kang Yang , Jiao Xiong , Lian Zhang , Wen-Juan Liang & Yong Shen
To cite this article: Chong-Kang Yang , Jiao Xiong , Lian Zhang , Wen-Juan Liang & Yong Shen
(2020): Two new dammarane-type triterpenoids from the steamed roots of Panaxꢀnotoginseng ,
Journal of Asian Natural Products Research, DOI: 10.1080/10286020.2020.1825391
To link to this article: https://doi.org/10.1080/10286020.2020.1825391
View supplementary material
Published online: 08 Oct 2020.
Submit your article to this journal
Article views: 3
View related articles
View Crossmark data
Full Terms & Conditions of access and use can be found at
https://www.tandfonline.com/action/journalInformation?journalCode=ganp20

JOURNAL OF ASIAN NATURAL PRODUCTS RESEARCH
https://doi.org/10.1080/10286020.2020.1825391
Two new dammarane-type triterpenoids from the
steamed roots of Panax notoginseng
a,b,c,d
a,b,c
a,b,c
e
Chong-Kang Yang
Yong Shen
, Jiao Xiong , Lian Zhang , Wen-Juan Liang and
a,b,c
a
Research & Development Department, Kunming Kangren Biotechnology CO., Ltd, Kunming 650033,
b
China; College of Agriculture and Biotechnology, Yunnan Agricultural University, Kunming 650201,
China; Research & Development Center for Functional Products, Yunnan Agricultural University,
Kunming 650201, China; Quality Management Department, Yunnan Baiyao Group Wuxi
Pharmaceutical CO., Ltd, Wuxi 214028, China; College of Food Science and Technology, Yunnan
c
d
e
Agricultural University, Kunming 650201, China
ABSTRACT
ARTICLE HISTORY
Two new dammarane-type triterpenoids, notoginsenoside SY3 (1)
and notoginsenoside SY4 (2), were isolated from the steamed
roots of Panax notoginseng. Their structures were determined
to be 3b, 12b, 20(S)-trihydroxy-27-anordammar-23(24)(E)-ene-3-O-
b-D-glucopyranosyl-(1!2)-b-D-glucopyranoside (1) and 3b, 12b,
Received 31 May 2020
Accepted 14 September 2020
KEYWORDS
Panax notoginseng;
dammarane-type
triterpenes; notoginsenoside
SY3; notoginsenoside SY4
2
0(S)-trihydroxy-25-methoxyldammar-23(24)(E)-ene-3-O-b-D-gluco-
pyranosyl-(1!2)-b-D-glucopyranoside (2) by IR, HRESIMS and
NMR experiments.
1. Introduction
Panax notoginseng (Burk.) F. H. Chen, a traditional Chinese medicine known as
Sanqi” in Chinese, is mainly distributed in Yunnan and Guangxi Provinces in China.
“
Besides, it refers to a vital herbal material resource for the drug industry of Yunnan
province [1]. The processed notoginseng, which is one critical form of this medicinal
plant, has been employed as a tonic for its blood cell-increasing function [2].
Ginsenosides 20(R/S)-Rh , Rk , Rh , 20(S/R)-Rg , Rk and Rg , the minor or even
1
3
4
3
1
5
trace components in the raw roots, were reported as the main constituents of the
processed roots [3, 4]. These mentioned saponins have gained prominence due to
CONTACT Wen-Juan Liang
liangwenjuan2007@163.com; Yong Shen
pharmsy@sina.com
Supplemental data for this article is available online at https://doi.org/10.1080/10286020.2020.1825391.
ß 2020 Informa UK Limited, trading as Taylor & Francis Group

2
C.-K. YANG ET AL.
Figure 1. Structures of compounds 1 and 2.
their potential pharmaceutical values of anticancer and enhancement on immune
[
5–9]. To be specific, ginsenoside Rg is the main component of ‘Shenyi capsule’
3
which has been used for the treatment of cancer in clinic [10]. In the present study,
two new saponins, notoginsenoside SY3 (1) and notoginsenoside SY4 (2) (Figure 1)
were isolated from the steamed roots of P. notoginseng and the isolation and struc-
tural elucidation were also described.
2. Results and discussion
Compound 1 was obtained as white amorphous powder, and its molecular formula
was assigned as C H O based on the HRESIMS ion peak at m/z 807.4508
4
1
68 14
þ
ꢀ1
[
M þ Na] . The IR spectrum exhibited absorption bands for hydroxyl (3408 cm ),
ꢀ1 ꢀ1
carbonyl (1658 cm ) and olefinic carbons (1453 cm ) groups in 1. Compound 1
1
3
displayed 41 resonances in the C NMR (DEPT) spectrum (Table 1) attributing to
seven methyls, ten methylenes, eighteen methines and six quaternary C-atoms, which
covered two anomeric carbons (d 105.6, 106.6) of sugar moieties, a pair of olefinic
C
1
carbons (d 134.1, 147.4) and one carbonyl carbon (d 198.3). In the H NMR spec-
C
C
trum (Table 1) of 1, seven singlet methyls [dH 0.85, 0.97, 1.06, 1.13, 1.30, 1.39 and
2
7
.26 (each 3H, s)], two olefinic protons [dH 6.37 (1H, d, J ¼ 16.0 Hz) and d 7.40-
H
0
.43 (1H, m)], and two anomeric protons at d 4.96 (1H, d, J ¼ 7.5 Hz, H-1 ), 5.41
H
0
0
(
d, J ¼ 7.2 Hz, H-1 ) were observed. The large coupling constants of the anomeric
protons were compatible with the b-configuration for all sugar moieties [11], which
were determined to be D-glucosyl moieties based on acidic hydrolysis experiment.
1
13
The H and C NMR spectral data (Table 1) of compound 1 were identical to those
of 3b, 12b, 20(S), 25- tetrahydroxydammar-23(24)(E)-ene-3-O-b-D-glucopyranosyl-
(
1!2)-b-D-glucopyranoside [12], except a ketone signal (d 198.3) appeared on the
C
side chain of 1 and one methyl signal was lost. Detailed analysis of the HMBC corre-
lations (Figure 2) from the olefinic protons at d 7.40-7.43 (1H, m) and 6.37 (1H, d,
H
J ¼ 16.0 Hz), and H -26 at d 2.26 to the ketone carbonyl carbon at d 198.3 indi-
3
H
C
cated the ketone carbonyl group bound to C-25. Moreover, the long-range correla-
0
00
0
tions between d 4.96 (H-1 ) and d 89.4 (C-3), d 5.41 (H-1 ) and d 83.9 (C-2 ) in
H
C
H
C

JOURNAL OF ASIAN NATURAL PRODUCTS RESEARCH
3
1
13
Table 1. H and C NMR spectral data for compounds 1 and 2.
1
ꢀ
2
Position
d
C
d
H
d
C
d
H
ꢁ
ꢁ
ꢁ
ꢁ
1
2
3
4
5
6
7
8
9
39.6
27.2
89.4
40.2
56.8
18.9
35.6
40.6
50.9
37.4
31.7
71.4
49.5
52.3
32.7
27.2
54.7
16.3
16.9
73.7
28.4
40.5
147.4
134.1
198.3
27.1
0.76-0.78 m, 1.51
1.84-1.86 m, 2.18
3.32 dt (10.2, 5.0)
40.6
27.2
91.3
40.9
57.5
19.2
35.9
41.0
51.3
37.9
31.9
72.2
51.3
52.6
32.1
27.2
54.7
16.2
16.7
74.4
27.2
40.2
128.2
138.4
76.6
26.2
26.3
28.4
16.8
17.1
50.6
1.03 , 1.71
1.72 , 1.85-1.87m
3.19
ꢁ
ꢁ
0.70 dd (11.8, 2.2)
0.78 d (10.1)
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
1.41 , 1.56
1.46 , 1.56
ꢁ
ꢁ
1.25 , 1.52-1.53 m
1.29 , 1.54
ꢁ
ꢁ
1.41
1.45
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
ꢁ
1.03-1.06 m, 1.56-1.57 m
1.23-1,25 m, 1.82
ꢁ
ꢁ
3.96
2.01
3.56
ꢁ
1.75-1.76 m
ꢁ
ꢁ
1.03 , 2.05-2.06 m
1.37 , 1.90-1.91 m
1.03-1.04 m, 1.54
ꢁ
ꢁ
ꢁ
1.32 , 1.97
2.38 td, (10.7, 7.1)
2.02-2.05 m
1.06 s
0.85 s
1.03 s
0.85 s
1.39 s
1.11 s
2.51 dd (14.0, 8.9), 2.89 d (14.3)
7.40-7.43 m
2.19-2.21 m, 2.35-2.37 m
5.73-5.77 m
5.47 d (16.0)
6.37 d, (16.0)
2.26 s
1.26 s
1.26 s
1.06 s
0.91 s
0.92 s
3.15 s
28.6
17.1
17.5
1.30 s
1.13 s
0.97 s
OCH
3
3
1
2
3
4
5
6
-O-Glc
0
105.6
83.9
78.8
72.1
78.8
63.3
4.96 d, (7.5)
4.28 t, (8.7)
105.4
81.0
77.9
71.9
78.4
63.1
4.45 d (7.4)
0
0
0
0
0
ꢁ
3.55
3.26
3.21
3.38
ꢁ
ꢁ
4.35
4.17
3.95
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
4.38 , 4.59 d (11.7)
3.64 , 3.81
Glc
1
2
3
4
5
6
"
"
"
"
"
"
106.6
77.7
78.5
72.1
78.6
63.2
5.41 d (7.2)
104.5
76.3
78.5
71.6
78.4
62.8
4.68 d (7.5)
ꢁ
ꢁ
4.17
4.28
4.35
3.95
3.23
3.62
3.36
3.38
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
4.51 , 4.52
3.66 , 3.86
ꢁ
Overlapped.
HMBC spectrum revealed the connectivity between the aglycone and the glycosyl
2
3,24
moieties (Figure 2). The geometry of the D
double bond was assigned as E owing
to the large coupling constant between H-23 and H-24 (J ¼ 16.0 Hz) [13]. The 20 R
configuration of 1 was confirmed by comparing the chemical shifts of C-17 and C-21
with literature data [11, 12, 14]. Accordingly, the structure of compound 1 was char-
acterized as 3b, 12b, 20(S)-trihydroxy-27-anordammar-23(24)(E)-ene-3-O-b-D-gluco-
pyranosyl-(1!2)-b-D-glucopyranoside, named as notoginsenoside SY3.
Compound 2 was isolated as a white amorphous powder. Its molecular formula
was determined to be C H O based on the pseudo-molecular ion at m/z 837.4971
4
3
74 14

4
C.-K. YANG ET AL.
Figure 2. Key HMBC correlations of compounds 1 and 2.
þ
[
M þ Na] . The IR spectrum of 1 displayed strong absorption bands at 3385, 2878
ꢀ
1
and 1637 cm due to hydroxyl, methoxy and olefinic carbonyl functional groups.
Compound 2 exhibited 43 resonances in the C NMR (DEPT) spectrum, including 8
13
methyls, a methoxyl (d 50.6), 10 methylenes, 18 methines, as well as 6 quaternary
C
13
C-atoms. The signals at d 138.4 and 128.2 in the C NMR spectrum indicated that
two olefinic carbons were present. In the H NMR spectrum (Table 1), seven singlet
C
1
methyls [d 0.85, 0.91, 0.92, 1.03, 1.06, 1.11, 1.26 ꢂ 2 (each 3H, s)], a methoxyl pro-
H
ton [d 3.15 (3H, s)], a pair of olefinic protons [dH 5.47 (1H, d, J ¼ 16.0 Hz), 5.73-
H
5
.77 (1H, m)] and two anomeric protons signals [dH 4.45 (d, J ¼ 7.4 Hz), 4.68 (d,
J ¼ 7.5 Hz)] were observed. As revealed from acid hydrolysis of 2, D-glucose was the
only sugar component. According to the coupling constant of the anomeric protons,
1
the b configuration for both of the glucose residues was verified [11]. The H and
1
3
C NMR spectra (Table 1) of compound 2 were similar to those of 3b, 12b, 20(S),
5-tetrahydroxydammar-23(24)(E)-ene-3-O-b-D-glucopyranosyl-(1!2)-b-D-glucopyr-
2
anoside [12], expect for the appearance of an additional methoxy signal (d 3.15 and
H
dC 50.6) in 2. In detailed inspection of the HMBC spectrum (Figure 2), a cross-peak
between d 3.15 (OCH ) and d 76.6 (C-25) indicated the methoxyl group was at C-
H
3
C
25 position. The long-range correlations between the aglycone and the glycosyl moi-
ety of 2 were the same as those of 1. The chemical shifts of C-17 and C-21 observed
at d 54.7 and 27.2, respectively, suggested that the configuration of C-20 was S [14].
C
The E configuration for the double bond between C-23 and C-24 was determined as
described previously. From the above data, the structure of compound 2 was estab-
lished to be 3b, 12b, 20(S)-trihydroxy-25-methoxyldammar-23(24)(E)-ene-3-O-b-D-
glucopyranosyl-(1!2)-b-D-glucopyranoside, named as notoginsenoside SY4.
3
. Experimental
.1. General experimental procedures
Optical rotations were determined on a Jasco model 1020 polarimeter (Horiba,
3
ꢀ
1
Tokyo, Japan). IR (KBr discs, cm ) spectra were recorded on a Bio-Rad FTS-135
spectrometer (Bio-Rad, Hercules, CA). 1D and 2D NMR were recorded on Bruker

JOURNAL OF ASIAN NATURAL PRODUCTS RESEARCH
5
DRX-500 or AVANCE 800 spectrometers (Bruker, Bremerhaven, Germany). High
resolution electrospray ionization mass spectrum (HRESIMS) were run on a VG
Spec3000 spectrometer (VG, Manchester, UK) and Waters AutoSpec Premier P776
(Waters, Milford, MA). D101 macroreticular resin (Tianjin Haiguang Chemical Co.,
Ltd, Haiguang, Tianjing, China), silica gel (200–300 mesh, Qingdao Makall Chemical
Company, Makall, Qingdao, China) and RP C18 silica gel (300–400 mesh, Merck
KGaA, Merck, Darmstadt, Germany) were used in the open column chromatography.
Gas chromatography (GC) was performed on the Agilent technologies 6890 N appar-
atus (Agilent, Santa Clara, USA). TLC was carried on the GF254 plates (Makall,
Qingdao, China). Analytical-grade reagents of chloroform (CHCl ), ethanol (EtOH),
3
and methanol (MeOH) were purchased from Kemiou Pure Chemical Co. Ltd.
(Kemiou Tianjin, China). Fractions were visualized by silica gel plates sprayed with
1
0% H SO ethanol solution followed by heating.
2 4
3
.2. Plant material
The air-dried roots of Panax notoginseng (Burk.) F. H. Chen were collected from
Wenshan county, Yunnan province, China, in December 2017, and authenticated by
Prof. Qing-Er Yang from South China Botanical Garden, Chinese Academy of
Sciences. The voucher specimen (No. YAU 2017-12-1) has been deposited in the
College of Agriculture and Biotechnology, Yunnan Agricultural University. The raw
plant sample was crushed into small pieces and then steamed at high temperature
ꢃ
(
120 C) and pressure (0.12 MPa) for 12 h, yielding the steamed roots of P. notogin-
seng preparation.
2
.3. Extraction and isolation
The steamed roots of P. notoginseng (15 kg) were extracted with 70% EtOH for three
times (3 h ꢂ 3) under reflux. After removal of the ethanol under reduced pressure, the
ethanolic extract was subjected to D101 resin column, eluting with H O and 90%
2
EtOH, successively. The EtOH eluent was combined and concentrated under reduced
pressure to give the total saponin fraction (2.1 kg), which was subjected to column
chromatography (CC) on silica gel (10 kg, 200–300 mesh) eluted with a gradient
CHCl -MeOH-H O (9:1:0.1!7:3:0.5) to provide eight fractions (Frs. A–G). Fr.D
3
2
(
418 g) was carried out on silica gel CC (CHCl -MeOH-H O, 8.5:1.5:0.15) to give six
3 2
fractions (Frs.D1 ꢀ D6). Fr.D3 (8.2 g) was repeatedly chromatographed over silica gel
(CHCl -MeOH-H O, 8.5:1.5:0.15) and Rp-18 columns (MeOH-H O, 8:2) to obtain 1
3 2 2
(5 mg) and 2 (6 mg).
2.3.1. Notoginsenoside SY3 (1)
White amorphous powder; Libermann-Burchard and Molish reactions were positive;
2
D
2
½
aꢄ þ9.78 (c 0.12, MeOH); IRꢀ
3408, 2968, 2945, 2878, 1658, 1453, 1388, 1367,
max
ꢀ
1
1
13
1
197, 1077, 1034 and 895 cm ; H NMR (800 MHz, Pyr) and C NMR (200 MHz,
þ
Pyr) spectral data are shown in Table 1; HRESIMS: m/z 807.4508 [M þ Na] (calcd
for C H O Na, 807.4501).
4
1
68 14

6
C.-K. YANG ET AL.
2.3.2. Notoginsenoside SY4 (2)
White amorphous powder; Libermann-Burchard and Molish reactions were positive;
2
D
2
½
aꢄ þ8.65 (c 0.19, MeOH); IRꢀ
3385, 2968, 2944, 2878, 1719, 1637, 1453, 1386,
max
ꢀ
1
1
13
1
199, 1076, 1035 and 893 cm ; H NMR (500 MHz, MeOD) and C NMR
(
[
125 MHz, MeOD) spectral data are shown in Table 1; HRESIMS: m/z 837.4971
þ
M þ Na] (calcd for C H O Na, 837.4971).
4
3
74 14
2
.4. Acid hydrolysis
The acid hydrolysis of new compounds and sugar determination were based on Gu’s
method by GC analysis [15]. Compounds 1 and 2 (each 5 mg) were hydrolyzed in
ꢃ
2
M HCl (5 ml) at 65 C for 6 h, respectively. The reaction mixture was extracted with
CHCl for three times (3 ꢂ 5 ml). The aqueous layer was neutralized with 2 M NaOH
3
and dried to produce a monosaccharide mixture. Then, a solution of the sugar mix-
ture in pyridine (2 ml) was added to L-cysteine methyl ester hydrochloride (1.5 mg)
ꢃ
and kept at 60 C for 1 h. After this, trimethylsilylimidazole (1.5 ml) was added to the
ꢃ
reaction mixture in an ice ꢀ water bath and kept at 60 C for 30 min. The mixture
was subjected to GC analysis, performed on an Agilent 6890 N gas chromatograph,
equipped with a 30 m ꢂ 0.32 mm HP-5 column and a H flame ionization detector
2
ꢃ
with the following conditions: column temperature, 200–260 C; programmed
increase, 10 C/min; carrier gas, N (1 ml/min); injector and detector temperature,
60 C; injection volume, 1 ll; and split ratio 1/50. The retention times for the
ꢃ
2
ꢃ
2
authentic samples (Sigma), after being treated in the similar manner, were 19.81 min
D-glucose) and 19.93 min (L-glucose), respectively. The peaks of the hydrolysate of
(
two compounds were detected at t 19.82 for D-glucose. Co-injection of each hydrol-
R
ysate with standard D-glucose gave single peaks.
Acknowledgments
The authors are grateful to the staff of analytical group at the State Key Laboratory of
Phytochemistry and Plant Resources in West China, Kunming Institute of Botany, Chinese
Academy of Sciences for the spectral data.
Disclosure statement
No potential conflict of interest was reported by the authors.
References
[1] T. Wang, R.X. Guo, G.H. Zhou, X.D. Zhou, Z.Z. Kou, F. Sui, C. Li, L.Y. Tang, and Z.J.
Wang, J. Ethnopharmacol. 188, 234 (2016).
[2] C.Z. Gu, J.J. Lv, X.X. Zhang, H. Yan, H.T. Zhu, H.R. Luo, D. Wang, C.R. Yang, M. Xu,
and Y.J. Zhang, Fitoterapia 103, 97 (2015).
[
[
3] P.Y. Liao, D. Wang, Y.J. Zhang, and C.R. Yang, J. Agric. Food Chem. 56, 1751 (2008).
4] D. Wang, P.Y. Liao, H.T. Zhu, K.K. Chen, M. Xu, Y.J. Zhang, and C.R. Yang, Food
Chem. 132, 1808 (2012).
[5] B. Chen, and X.B. Jia, Molecules 18, 8109 (2013).

JOURNAL OF ASIAN NATURAL PRODUCTS RESEARCH
7
[
[
6] B. Chen, Y.P. Shen, D.F. Zhang, J. Cheng, and X.B. Jia, Nat. Prod. Res. 27, 2351 (2013).
7] S. Sun, L.W. Qi, G.J. Du, S.R. Mehendale, C.Z. Wang, and C.S. Yuan, Food Chem. 125,
1299 (2011).
[
[
8] S. Sun, C.Z. Wang, R. Tong, X.L. Li, A. Fishbein, Q. Wang, T.C. He, W. Du, and C.S.
Yuan, Food Chem 118, 307 (2010).
9] D.F. Toh, D.N. Patel, E.C.Y. Chan, A. Teo, S.Y. Neo, and H.L. Koh, Chin. Med. 6, 4
(
2011).
10] P. Lu, W. Su, Z-h Miao, H-r Niu, J. Liu, and Q-l Hua, Chin. J. Integr. Med. 14, 33
2008).
11] R.G. Teng, H.Z. Li, J.T. Chen, D.Z. Wang, Y.N. He, and C.R. Yang, Magn. Reson.
Chem. 40, 483 (2002).
12] J.R. Wang, L.F. Yau, R. Zhang, Y. Xia, J. Ma, H.M. Ho, P. Hu, M. Hu, L. Liu, and Z.H.
Jiang, J. Agric. Food Chem. 62, 2558 (2014).
13] T.L. Tran, Y.R. Kim, J.L. Yang, D.R. Oh, T.T. Dao, and W.K. Oh, Bioorg. Med. Chem.
[
[
[
[
(
2
2, 499 (2014).
[
[
14] H. Yang, J.Y. Kim, S.O. Kim, Y.H. Yoo, and S.H. Sung, J. Ginseng Res. 38, 194 (2014).
15] C.Z. Gu, J.J. Lv, X.X. Zhang, Y.J. Qiao, H. Yan, Y. Li, D. Wang, H.T. Zhu, H.R. Luo,
C.R. Yang, M. Xu, and Y.J. Zhang, J. Nat. Prod. 78, 1829 (2015).