Two new compounds from Potentilla multicaulis Bunge

Article abstract of DOI:10.1080/14786419.2012.740037

Two new compounds (triterpenoid saponin and heterocyclic compound), 2α,3β, 19α,23,30-pentahydroxyurs-12-en-28-oic acid-28-O-β-d-glucopyranosyl ester (1) and N-hydroxyl-hexahydroazepin-2,4- diones (2), with 11 known compounds, picein (3), (7S,8R)- dihydrod

Full text of DOI:10.1080/14786419.2012.740037

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Two new compounds from Potentilla
multicaulis Bunge
a
a
a
a
Lingyun Jia , Jing Wang , Chongning Lv , Tanye Xu , Lihua He
b
a
a
,
Yan Dong & Jincai Lu
a
School of Traditional Chinese Materia Medica, Shenyang
Pharmaceutical University , Box 79, Wenhua Road 103, Shenhe
District, Shenyang , Liaoning Province , 110016 , PR China
b
School of Medical Devices, Shenyang Pharmaceutical University ,
Shenyang , Liaoning Province , 110016 , PR China
Published online: 13 Nov 2012.
To cite this article: Lingyun Jia , Jing Wang , Chongning Lv , Tanye Xu , Lihua He , Yan Dong &
Jincai Lu (2013) Two new compounds from Potentilla multicaulis Bunge, Natural Product Research:
Formerly Natural Product Letters, 27:15, 1361-1365, DOI: 10.1080/14786419.2012.740037
To link to this article: http://dx.doi.org/10.1080/14786419.2012.740037
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Natural Product Research, 2013
Vol. 27, No. 15, 1361–1365, http://dx.doi.org/10.1080/14786419.2012.740037
Two new compounds from Potentilla multicaulis Bunge
a
Lingyun Jia , Jing Wang , Chongning Lv , Tanye Xu , Lihua He , Yan Dong and
a
a
a
b
a
a
Jincai Lu *
a
School of Traditional Chinese Materia Medica, Shenyang Pharmaceutical University, Box 79, Wenhua
Road 103, Shenhe District, Shenyang, Liaoning Province 110016, PR China; School of Medical Devices,
Shenyang Pharmaceutical University, Shenyang, Liaoning Province 110016, PR China
b
(
Received 1 July 2012; final version received 16 September 2012)
Two new compounds (triterpenoid saponin and heterocyclic compound), 2a,3b,
19a,23,30-pentahydroxyurs-12-en-28-oic acid-28-O-b-D-glucopyranosyl ester (1)
and N-hydroxyl-hexahydroazepin-2,4-diones (2), with 11 known compounds,
0
picein (3), (7S,8R)-dihydrodehydrodiconiferyl alcohol-9 -O-b-D-glucopyranoside (4),
(
þ)-1-hydroxy-2-epipinoresinol-1-b-D-glucoside (5), (þ)-1-hydroxypinoresinol-1-b-
0
D-glucoside (6), (þ)-1-hydroxypinoresinol-4 -b-D-glucoside (7), schaftside (6-C-b-D-
glucopyranosyl-8-C-a-L-arabinosyl apigenin) (8), isoschaftside (6-C-a-L-arabinosyl-8-
C-b-D-glucopyranosyl apigenin) (9), isorhamnetin-3-O-b-D-glucopyranoside (10),
quercetin-3-O-b-glucuronide (11), 8-O-methylherbacetin-3-O-sophoroside (12) and
kaempferol (13), were isolated from Potentilla multicaulis Bunge. The structure of the
compounds was elucidated by chemical and spectral evidence.
Keywords: Potentilla multicaulis Bunge; triterpenoid saponin; heterocyclic
compound; structural elucidation
1
. Introduction
Potentilla multicaulis Bunge is widely distributed in the northeast province of China (Institute of
Botany, 1972). The whole plant of P. multicaulis Bunge has been used as a Chinese folk
medicine to treat diabetes as a substitute of Potentilla discolor Bunge, which is a well-known
treatment for this disease in China (Hong, Li, & Pu, 2007; Liu, Yan, & Xu, 2003; Ma & Wen,
2
002). The genus Potentilla L., which belongs to the Rosaceae family, includes more than 200
species of plants widely distributed in the north temperate zone, boreal and alpine areas, and a
few species near the equator. The aim of our work was to carry out the chemical study of P.
multicaulis Bunge that has never been reported before to provide the chemical basis for P.
multicaulis Bunge as a substitute of P. discolor Bunge. We had reported the phytochemical
studies of this plant before (He, Hua, Zhang, & Lu, 2009). This study has also focused on
investigating the remaining constituents of this herb medicine. Here, we report our recent
investigation resulting in isolation and structural characterisation of two new compounds
(
1 and 2).
*
Corresponding author. Email: jincailu@yahoo.com.cn
q 2013 Taylor & Francis

1
362
L. Jia et al.
2
. Results and discussion
The chemical studies of 70% ethanol extracts of P. multicaulis Bunge, by different
chromatographic techniques, afforded compounds 1 and 2. Compounds 1 and 2 were new
compound and were identified by 1D, 2D NMR spectroscopy (Figure 1) and high-resolution
(
comparison of their NMR data with those reported in the literature as picein (3), (7S,8R)-
HR)-ESI-MS analysis, whereas the other compounds are known compounds identified by
0
dihydrodehydrodiconiferyl alcohol-9 -O-b-D-glucopyranoside (4), (þ)-1-hydroxy-2-epipinor-
esinol-1-b-D-glucoside (5), (þ)-1-hydroxypinoresinol-1-b-D-glucoside (6), (þ)-1-hydroxypi-
0
noresinol-4 -b-D-glucoside (7), schaftside (6-C-b-D-glucopyranosyl-8-C-a-L-arabinosyl
apigenin) (8), isoschaftside (6-C-a-L-arabinosyl-8-C-b-D-glucopyranosyl apigenin) (9),
isorhamnetin-3-O-b-D-glucopyranoside (10), quercetin-3-O-b-glucuronide (11), 8-O-methyl-
herbacetin-3-O-sophoroside (12) and kaempferol (13).
Compound 1 was obtained as white needles. The molecular formula was established as
þ
C H O by the positive-ion HR-ESI-MS at m/z: 705.3751 [M þ Na] (calcd. 705.3820). The IR
3
6 58 12
21
21
spectrum also released the absorption of hydroxyl (3422cm ), carbonyl (1732 cm ) and double
21
13
band (1636 cm ). The C NMR spectrum showed 36 carbon signals, of which 30 were assigned to
3
the triterpenoid aglycone and 6 to the sugar moieties. The five sp carbons at d 14.4, 17.6, 17.6, 24.1
2
1
and 27.4, and the two sp carbons at d 128.7 and 138.8, coupled with information from the H NMR
fivemethylprotonsingletsatd1.08, 1.14, 1.23, 1.55and1.60, andabroadsinglevinyl protonsignal
(
1
3
at d 5.54), indicated that the aglycone had an urs-12-ene skeleton. Furthermore, the C NMR
spectrum showed a carbonyl signal at d 177.0 (C-28). These together with comparison of chemical
shifts for the aglycone region of nigaichigoside F (Zhang, Peng, & Wang, 2001), only chemical
1
shifts were observed at C-19 (þ1.3 ppm), C-20 (þ6 ppm), C-21 (þ4.4 ppm) and C-30
(
þ48.1ppm), which enabled the identification of the aglycone in 1 as 2,3,19,23, 30-pentahydroxy-
1
preparation layer chromatography (PTLC). The absolute configuration of the sugar was affirmed by
2-ursen. After 1 was hydrolysed inacidic condition, D-glucose was isolated fromthe water layer by
1
13
optical rotation. The H and C NMR spectra of 1 revealed signals assignable to b-glucopyranosyl
moiety with the anomeric proton signals at d 6.30 (1H, d, J ¼ 8.4 Hz, Glc-H-1), and the
Figure 1. Structures of compounds isolated from P. multicaulis Bunge.

Natural Product Research
1363
corresponding anomeric carbon signals at d 95.9 (Glc-C-1), which were also observed in the
heteronuclear singular quantum correlation (HSQC) experiment. The glucopyranosyl moiety bond
connected between d 177.0 (C-28)and d 6.30(1H, d, J ¼ 8.4 Hz, Glc-H-1), d 2.93(1H, s, H-18),
C
H
H
1
d 1.88 (1H, ddd, J ¼ 4.2, 13.2Hz, H-22). Other structural fragments were confirmed by H NMR,
H
3
1
C NMR and 2D NMR (HSQC and heteronuclear multiple-bond correlation (HMBC)) as showed
in Table 1. Thus, the structure of compound 1 was determined to be 2a,3b,19a, 23,30-
pentahydroxyurs-12-en-28-oic acid-28-O-b-D-glucopyranosyl ester (Figure 2).
Compound 2 was obtained as colourless needles, the molecular formula of 2 was determined
þ
as C H O N by the HR-ESI-MS at m/z: 144.0657 [M þ H] (calcd. 144.0655), as well as from
6
9 3
2
1
its NMR spectroscopic data. The IR spectrum suggested the presence of a hydroxyl (3423 cm
)
2
1
1
and two carbonyl groups (1716 and 1632 cm ). Eight protons emerged in the H NMR
spectrum at d 2.10 (2H, m), 2.47 (2H, t, J ¼ 7.8 Hz), 3.52 (2H, t, J ¼ 7.2 Hz) and 4.09 (2H, s).
Taking into consideration of the two carboxyl (d 171.6 and 176.7) and other four carbon signals
d 17.9, 30.4, 44.2 and 48.2) found in the C NMR spectra, the molecule was believed to contain
NZCH ZCH ZCH ZCOZ and ZCOZCH ZCOZ fragments. Simultaneously, the proton
1
3
(
2
2
2
2
2
1
signal at d 8.45 (1H, br s), in addition to the IR spectrum at 3423 cm (ZOH), revealed the
presence of one NZOH fragment. The complete molecule was further demonstrated by 2D
NMR (HSQC and HMBC) experiments. With all the above evidence, the structure of 2 was
determined to be N-hydroxyl-hexahydroazepin-2,4-diones (Figure 2).
3
3
. Experimental
.1. General
Melting points were measured on a General Experimental Procedures X-4 purchased from
Beijing Tech Instrument Co. Ltd (Beijing, China), and are uncorrected. IR spectra were recorded
using a Bruker IFS-55 IR spectrometer with KBr discs. HR-ESI-MS were recorded on a Bruker
micrOTOF-Q MS spectrometer. NMR spectra were recorded on a Bruker ARX-300 and Bruker
AV-600 NMR spectrometers, with tetramethylsilane (TMS) as the internal standard. Silica
GF254 for thin layer chromatography (TLC) and silica gel (200–300 mesh) for column
chromatography were obtained from Qingdao Marine Chemical Company (Qingdao, People’s
Republic of China). Semi-preparative high performance liquid chromatography (HPLC) was
carried out on a server side include (SSI) instrument consisting of a series pump and a Model 500
Figure 2. Structures and key HMBC correlations of compounds 1 and 2.

1
364
L. Jia et al.
UV detector. A YMC*Gel ODS-A was purchased from YMC Co. Ltd (YMC Karasume-Gojo
Bldg., Japan). Reagents used were of analytical grade and purchased from Yuwang Group
(
Shandong, People’s Republic of China).
3
.2. Plant material
The whole herb of P. multicaulis Bunge was collected from Liaoning Province of China
in August 2006 and was identified by Professor Qishi Sun (Department of Medicinal
Plant, College of Traditional Chinese Medicine of Shenyang Pharmaceutical University). A
voucher specimen (Voucher no.: 002155) has been deposited in the Herbarium of Shenyang
Pharmaceutical University.
3
.3. Extraction and isolation
The dry whole plant (10.8 kg) of P. multicaulis Bunge was extracted three times with 100 l of
0% ethanol for 1 h each time under reflux, and the solution was concentrated under reduced
7
pressure. The resulting extract (1442 g) was subjected to a column (f 15 £ 200 cm) containing
macroporous resin (AB8) and eluted with H O, 30%, 50%, 70%, 95% aqueous EtOH in six
2
volumes of resin, successively. Evaporation of the solvent under reduced pressure delivered
these fractions (frs). The viscous mass (35 g) of 95% aqueous EtOH was chromatographed on
3
1
50 g silica gel column (f 10 £ 120 cm), eluting with (P.E.)-EtOAc (50:1 ! 0:1) (20:1, 15:1,
0:1, 5:1, 2:1, 1:1) to obtain 12 frs. Fr. 10 was purified by Sephadex LH-20 eluted with MeOH:
H O (6:4) to afford a further 10 fractions (Fr. 10-1 to 10-10). Fr. 10-4 was further recrystallised
2
to obtain compound 2 (30 mg). The concentrated viscous mass (292 g) of 70% aqueous EtOH
was subjected to a column (f 10 £ 200 cm) containing silica gel 60, and then eluted with a linear
gradient system of CHCl ZMeOH (100:1 ! 0:1) in 10 volumes of resin, and 500 mL of each fr.
3
was collected to give 20 frs. Fr. 8 was further repeatedly subjected to Sephadex LH-20, modified
decylsilyl silicion (MDS), octa decylsilyl silicion (ODS) chromatography and semi-preparative
HPLC with various solvent systems to yield four known compounds 4 (125.4 mg), 5 (10 mg), 6
(
with MeOHZH O (1:9–4:6) and further recrystallised to afford compound 1 (9 mg). Fr. 10 was
23.3 mg), 7 (11 mg). Fr. 9 was repeatedly subjected to Sephadex LH-20, ODS chromatography
2
also repeatedly subjected to Sephadex LH-20, MDS, ODS chromatography and preparative
HPLC with various solvent systems to yield six known compounds 8 (13 mg), 9 (7 mg), 10
(10 mg), 11 (15 mg), 12 (23 mg), 13 (11 mg). The dissolved part of the concentrated viscous
mass of 30% aqueous EtOH was isolated by Sephadex LH-20 and then eluted with gradient
MeOHZH O (1:9–1:0) to give 11 frs. Fr. 4 was further purified by RP C-18 silica gel column (f
2
3
obtain compound 3 (27 mg).
£ 60 cm), eluting with MeOHZH O (2:8) to obtain 10 frs. Fr. 5 was further recrystallised to
2
4
. Acid hydrolysis of compound 1
An aqueous solution of 1 (5 mg) in 1 M HCl (2 mL) was heated at 808C for 2 h. After the pH
being modified to 7.0 with NaHCO solution, the reaction mixture was extracted with CHCl
3
3
(
eluent to yield an aglycone. The water layer was concentrated and subjected to PTLC (EtOAc/
3 £ 4 mL). CHCl part was evaporated and subjected to PTLC, using CHCl /CH OH (10:1) as
3
3
3
CH OH/H O ¼ 7:3:0.4) to yield the sugar which were identified by TLC (CHCl /CH OH/
3
2
3
3
2
D
5
H O ¼ 16:9:1) with authentic samples and [a] as follows: D-glucose ½aꢀ þ 9.68 (c 0.12,
2
D
H O), spot was detected by spraying with EtOHZH SO –anisaldehyde (17:2:1) followed by
2
2
4
heating, D-glucose were identified.

Natural Product Research
1365
5
. Data of compound 1
25
White needles; m.p. 214–2158C; ½aꢀ 211.78 (c 0.22, MeOH); IR nmax (film on KBr): 3422,
D
2
1
927, 1732, 1636, 1074, 619 cm ; showed a quasi-molecular ion peak at m/z 705.3751
2
þ
1
[
C D N), d 1.38 (1H, m, H-1), 2.30 (1H, dd, 4.2, 12.0, H-1); 4.24 (1H, overlapped, H-2); 4.18
M þ Na] (calcd. for C H O Na, 705.3820) in the HR-FAB-MS; H NMR (600 MHz, in
5 5 H
3
6 58 12
(
(
(
1H, overlapped, H-3); 1.83 (1H, d, 11.4, H-5); 1.45, 1.69 (2H, m, H-6); 1.45 (1H, m, H-7), 1.75
1H, ddd, 3.6, 11.2, H-7); 2.05 (1H, m, H-9); 1.92 (1H, m, H-10); 2.14, 1.58 (2H, m, H-11); 5.54
1H, s, H-12); 1.18 (1H, dd, 3.6, 10.2, H-15), 2.45 (1H, m, H-15); 2.00 (1H, m, H-16), 3.13 (1H,
ddd, 4.2, 13.2, H-16); 2.93 (1H, s, H-18); 1.45 (1H, m, H-20); 2.42, 1.38 (2H, m, H-21); 1.88
1H, ddd, 4.2, 13.2, H-22), 2.14 (1H, m, H-22); 4.18 (1H, overlapped, H-23), 3.70 (1H, d,10.8,
H-23); 1.08 (3H, s, H-24); 1.23 (3H, s, H-25); 1.23 (3H, s, H-26); 1.60 (3H, s, H-27); 1.55 (3H, s,
(
0
H-29); 3.98 (1H, dd, 1.8, 10.8, H-30), 4.23 (1H, overlapped, H-30); 6.30 (1H, d, 8.4, H -1); 4.22
0
0
0
0
(
1H, m, H -2); 4.05 (1H, m, H -3); 4.36 (1H, t, 9.6, H -4); 4.30 (1H, t, 9.6, H -5); 4.40 (1H, dd,
0 0
.2, 12.0, H -6), 4.47 (1H, dd, 2.4, 12.0, H -6). C NMR (150 MHz, in C D N), d 48.0 (C-1);
5 5 C
9.0 (C-2); 78.4 (C-3); 43.6 (C-4); 48.0 (C-5); 18.7 (C-6); 33.2 (C-7); 40.6 (C-8); 47.9 (C-9);
8.4 (C-10); 24.2 (C-11); 128.7 (C-12); 138.8 (C-13); 42.2 (C-14); 29.2 (C-15); 26.3 (C-16);
8.6 (C-17); 54.3 (C-18); 74.0 (C-19); 48.2 (C-20); 22.3 (C-21); 37.7 (C-22); 66.6 (C-23); 14.4
13
4
6
3
4
(
(
C-24); 17.6 (C-25); 17.6 (C-26); 24.1 (C-27); 177.0 (C-28); 27.4 (C-29); 64.8 (C-30); 95.9
0
0
0
0
0
0
C-1 ); 74.1 (C-2 ); 79.3 (C-3 ); 71.2 (C-4 ); 79.0 (C-5 ); 62.3 (C-6 ).
6
. Data of compound 2
Colourless needles; m.p. 147–1498C; IR nmax (film on KBr): 3423, 2964, 1716, 1660, 1478,
1
2
397, 1342, 1264, 1002, 948, 873, 777, 658, 559, 511 cm ; showed a quasi-molecular ion peak
1
þ
1
at m/z 144.0657 [M þ H] (calcd. 144.0655) in the HR-FAB-MS; H NMR (600 MHz, in
DMSO-d ), d 8.45 (1H, br s, H-1); 4.09 (2H, s, H-3); 2.47 (2H, t, J ¼ 7.8 Hz, H-5); 2.10 (2H, m,
6
H
1
3
H-6); 3.52 (2H, t, J ¼ 7.2 Hz, H-7). C NMR (150 MHz, in DMSO-d ), d 171.6 (C-2); 44.2
6
C
(
C-3); 176.7 (C-4); 30.4 (C-5); 17.9 (C-6); 48.2 (C-7).
Supplementary material
Spectral data relating to this article are available online.
Acknowledgements
The authors are grateful to the analytical detective centre, Shenyang Pharmaceutical University, for
recording NMR, UV, IR, ESIZMS and HR-ESI-MS spectra.
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