New triterpenoids from the leaves of Cyclocarya paliurus

Article abstract of DOI:10.1055/s-0031-1280411

Six new triterpenoids including four new secodammarane triterpenoid glycosides (14), an epoxydammarane triterpenoid glycoside (5), and a new secodammarane triterpenoid (6) were isolated from the ethanolic extract of the leaves of Cyclocarya paliurus. The structures of these compounds were elucidated by spectroscopic analysis methods. Compounds 16 were evaluated for their inhibitory activities against α-glucosidase, lipase, DPP-IV, and aldose reductase. Georg Thieme Verlag KG Stuttgart - New York.

Full text of DOI:10.1055/s-0031-1280411

2
90 Letters
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7
8
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Chen YY, Wang XN, Fan CQ, Yin S, Yue JM. Swiemahogins A and B, two
novel limoids from Swietenia mahogani. Tetrahedron Lett 2007; 48:
480–7484
Luo J, Wang JS, Huang XF, Luo JG, Kong LY. Chukvelutilides A–F, phrag-
malin limonoids from the stem barks of Chukrasia tabularis var. veluti-
na. Tetrahedron 2009; 65: 3425–3431
Wu J, Xiao Q, Zhang S, Li X, Xiao ZH, Ding HX, Li QX. Xyloccensins Q–V, six
new 8,9,30-phragmalin ortho ester antifeedants from the Chinese
mangrove Xylocarpus granatum. Tetrahedron 2005; 61: 8382–8389
Abdelgaleil SAM, Doe M, Morimoto Y, Nakatani M. Rings B, D-seco limo-
noids from the leaves of Swietenia mahogany. Phytochemistry 2006;
7
New Triterpenoids from the Leaves of
Cyclocarya paliurus
1
1
1
2
1
Shuai Li , Baosong Cui , Quan Liu , Li Tang , Yongchun Yang ,
1
1
Xiangju Jin , Zhufang Shen
1
State Key Laboratory of Bioactive Substance and Function of Natural
Medicines, Institute of Materia Medica, Chinese Academy of
Medical Sciences and Peking Union Medical College, Beijing, P.R.
China
6
7: 452–458
1
0 Wikberg JES, Rasoanaivo P, Benoit R, Razafimahefa AS. Novel com-
pounds and pharmaceutical preparations from Neobeguea mahafalen-
sis extracts and their use for treatment of sexual dysfunction. US Patent
2
Division of Experimental Therapeutics, Department of Medicine,
Columbia University, New York, New York, USA
1
45996A2; 2008
1
1
1
1 Chen PY. Meliaceae. In: Chen SK, editor. Flora reipublicae popularis si-
nicae (Zhongguo Zhiwu Zhi), Volume 13. Beijing: Science Press; 1997:
Abstract
4
7–48
!
2 Editorial Committee of the Administration Bureau of Traditional Chinese
Medicine. Chinese materia medica (Zhonghua Bencao), Volume 5.
Shanghai: Shanghai Science and Technology Press; 1999: 31–32
3 Luo J, Wang JS, Luo JG, Wang XB, Kong LY. Chukvelutins A–C, 16-nor-
phragmalin limonoids with unprecedented skeletons from Chukrasia
tabularis var. velutina. Org Lett 2009; 11: 2281–2284
4 Zhang CR, Yang SP, Zhu Q, Liao SG, Wu Y, Yue JM. Nortriterpenoids from
Chukrasia tabularis var. velutina. J Nat Prod 2007; 70: 1616–1619
5 Sun YQ, Guo YW. Gymonorrhizol, an unusual macrocyclic polydisulfide
from the Chinese mangrove Bruguiera gymnorrhiza. Tetrahedron Lett
Six new triterpenoids including four new secodammarane triter-
penoid glycosides (1–4), an epoxydammarane triterpenoid glyco-
side (5), and a new secodammarane triterpenoid (6) were iso-
lated from the ethanolic extract of the leaves of Cyclocarya paliu-
rus. The structures of these compounds were elucidated by spec-
troscopic analysis methods. Compounds 1–6 were evaluated for
their inhibitory activities against α-glucosidase, lipase, DPP‑IV,
and aldose reductase.
1
1
2
004; 45: 5533–5535
1
1
1
1
2
2
6 Wang JD, Guo YW. Agallochaols A and B, two new diterpenes from the
Chinese mangrove Excoecaria agallocha L. Helv Chim Acta 2004; 87:
Key words
Cyclocarya paliurus · Juglandaceae · triterpenoid · secodam-
marane type · biological activity
2
829–2833
7 Wang JD, Zhang W, Li ZY, Xiang WS, Guo YW, Krohn K. Elucidation of ex-
cogallochaols A–D, four unusual diterpenoids from the Chinese man-
grove Excoecaria agallocha. Phytochemistry 2007; 68: 2426–2431
8 Liu HL, Huang XY, Dong ML, Xin GR, Guo YW. Piperidine alkaloids from
Chinese mangrove Sonneratia hainanensis. Planta Med 2010; 76: 920–
Supporting information available online at
http://www.thieme-connect.de/ejournals/toc/plantamedica
9
22
9 Byon JCH, Kusari AB, Kuseti J. Protein-tyrosine phosphatase-1B acts as a
negative regulator of insulin signal transduction. Mol Cell Biochem
Cyclocarya paliurus (Batal.) Iljinsk (Juglandaceae) is a Chinese en-
demic plant which has been used to treat hypertension and dia-
betes [1,2] and is known to have a hypolipemic effect [3]. Some
dammarane-, oleanane-, and ursane-type triterpenes, flavo-
noids, and phenolic acids were reported from this plant [4–18].
Phytochemical investigation of its leaves was undertaken to as-
sess the chemical and biological diversity. This report describes
the isolation and characterization of six new triterpenoids from
1
998; 182: 101–108
0 Lammer C, Wagerer S, Saffrich R, Mertens D, Ansorge W, Hoffmann I. The
cdc25B phorsphatase is essential for the G2/M phase transition in hu-
man cell. J Cell Sci 1998; 111: 2445–2453
1 Wignot TM, Stewart RP, Schray KJ, Das S, Sipos T. In vitro studies of the
effects of HAART drugs and excipients on activity of digestive enzymes.
Pharm Res 2004; 21: 420–427
received April 13, 2011
revised October 26, 2011
accepted November 7, 2011
"
the leaves of C. paliurus (l Fig. 1). Chemical and spectroscopic
methods were used to elucidate the structures of the new com-
pounds.
Bibliography
The HR‑ESI‑MS of 1 exhibited the molecular formula C37H62O .
9
DOI http://dx.doi.org/10.1055/s-0031-1280403
Published online December 1, 2011
Planta Med 2012; 78: 286–290
1
The H NMR spectrum of 1 showed seven quaternary methyls, a
pair of coupled olefinic protons at δ 5.69 and 5.44, whose cou-
pling constant (J = 16.0 Hz) showed that the configuration of the
double bond was E, terminal methylene olefinic protons at δ 4.83
and 4.72, and an oxymethine proton at δ 4.13, in addition to par-
tially overlapped mutiplets of methylenes and methines protons
between δ 0.78 and δ 2.70. These data were used to assign the tri-
terpenoid aglycone skeleton. The presence of an anomeric proton
of glycosyl at δ 4.40 as well as other oxymethine protons and one
methyl doublet of glycosyl suggested that 1 was a triterpenoid
©
Georg Thieme Verlag KG Stuttgart · New York ·
ISSN 0032‑0943
Correspondence
Prof. Dr. Yue-Wei Guo
State Key Laboratory of Drug Research
Shanghai Institute of Materia Medica
Chinese Academy of Sciences
Zu Chong Zhi Rd. 555
Zhangjiang, Hi-Tech Park
Shanghai 201203
Peopleʼs Republic of China
Phone: + 862150805813
Fax: + 862150805813
1
3
saponin. The C NMR spectra of 1 showed 37 carbon signals in-
cluding a triterpenoid skeleton as well as glycosyl and methoxy
"
moieties (l Table 1). The seven degrees of unsaturation of 1 re-
ywguo@mail.shcnc.ac.cn
quired by the molecular formula included one carboxyl, two
double bonds, and one ring of glycosyl, so that three degrees of
unsaturation remained, confirming that 1 possessed an unusual
triterpenoid skeleton with a tricyclic parent nucleus, which was
Li S et al. New Triterpenoids from… Planta Med 2012; 78: 290–296

Letters 291
Fig. 1 Structures of compounds 1–6.
finally established by detailed analyses of 2D NMR spectra of 1. A
further comparison of the NMR data of 1 with those of the related
compounds in the literature [6] indicated that 1 was in good
agreement with (23E)-(12R,20S)-20-hydroxy-3,4-secodam-
mara-4(28),23-dien-3-oic acid 12-O-β-D-quinovopyranoside ex-
cept for the data of C-25, which was methoxylated in 1. HMBC
unit was also linked at a C-12 position. The stereochemistry at C-
12 and C-20 were determined to be R and S, respectively, on the
1
3
basis of comparison to the C NMR chemical shift data and opti-
cal rotation for analogous 3,4-secodammaranes [6,19]. The struc-
ture of 3 was deduced as (23E)-(12R,20S)-12,20-dihydroxy-25-
methoxy-3,4-secodammara-4(28),23-dien-3-oic acid 12-O-α-L-
arabinopyranoside, and 3 was named cyclocarioside F.
"
correlations from H -OCH (3.08) to C-25 (75.1) (l Fig. 2) con-
3
3
firmed that the methoxy group was linked to position C-25.
HMBC correlations from H-1′ (4.40) to C-12 (75.5) (l Fig. 2) indi-
cated that the glycosyl was located at C-12. The absolute configu-
The HR‑FAB‑MS of 4 exhibited the molecular formula C38H64O9.
The NMR spectrum of 4 showed similarities to that of 3, except
that the NMR signals due to the proton of 3-carboxyl of 3 were
replaced by signals attributable to an ethyl unit (δH 4.06 and
"
rations at C-12 and C-20 were determined to be R and S, respec-
1
3
tively, on the basis of comparison to the C NMR chemical shift
and optical rotation data for analogous 3,4-secodammaranes [6,
1.22; δ 60.6 and 14.5). This suggested that 4 was a derivative of
C
1
3
3 containing an ethyl ester unit. A comparison of the C NMR da-
ta between 3 and 4 (l Table 2) indicated that the C-3 of 4 was
"
19]. The stereochemistry of C-5 and C-10 were determined on
the basis of the natural dammarane-type triterpene biosynthetic
pathway [20]. Ultimately, the structure of 1 was deduced as
shielded by ΔδC 1.1 ppm; this suggested that the location of the
ethyl unit was also at the C-3-carboxyl of 4. Therefore, the struc-
ture of 4 was deduced as (23E)-(12R,20S)-12,20 ‑dihydroxy-25-
methoxy-3,4-secodammara-4(28),23-dien-3-oic acid ethyl ester
12-O-α-L-arabinopyranoside, and 4 was named cyclocarioside G.
(23E)-(12R,20S)-12,20-dihydroxy-25-methoxy-3,4-secodam-
mara-4(28),23-dien-3-oic acid 12-O-β-D-quinovopyranoside,
and 1 was named cyclocarioside D.
The HR‑FAB‑MS of 2 exhibited the molecular formula C39H66O9.
The NMR spectrum of 2 showed similarities to that of 1, except
that the NMR signals due to the proton of 3-carboxyl of 1 was re-
placed by signals attributable to an ethyl unit (δH 4.02 and 1.20;
δC 60.4 and 14.6). This suggested that 2 was a derivative of 1 con-
taining an ethyl ester unit at the 3-carboxyl. In the HMBC spec-
trum of 2, long-range heteronuclear correlations from δH 4.02 to
δC 175.6 (C-3) proved the location of the ethyl unit at C-3. Ulti-
mately, the structure of 2 was deduced as (23E)-(12R,20S)-20-hy-
droxy-25-methoxy-3,4-secodammara-4(28),23-dien-3-oic acid
ethyl ester 12-O-β-D-quinovopyranoside, and 2 was named cy-
clocarioside E.
The HR‑FAB‑MS of 5 exhibited the molecular formula C43H72O13.
The NMR spectrum of 5 showed that it was a triterpenoid glyco-
side with a dammarane triterpenoid aglycone skeleton and two
"
sugars (l Table 3). The configuration of the hydroxyl group at C-
3
3 was deduced to be α-positioned by a small JH,H coupling con-
stant. The NMR signals of protons and protonated carbons were
assigned by the HSQC experiment. Further comparison of the
NMR data of 5 with the related compound cyclocarioside I in the
literature [4] indicated that the NMR data of 5 were in good
agreement with those of (20S,24R)-epoxydammarane-(3β,12β)-
25-hydroxyl-12-O-β-D-quinovopyranosyl-3-O-α-L-arabinofura-
noside except for the data of C-5′ and C-3. Substitution patterns
and position of C-5′ and C-3 were defined in the HMBC spectrum;
the correlations from H-3 (δH 3.26) to C-1′ (106.0) and H-1′ (δH
4.93) to C-3 (79.6) revealed the linkage position of arabinofura-
nose at C-3; the correlations from H-5′ (4.10, 4.26) to C=O
(170.8) of the acetyl group confirmed that the acetyl group was
attached to H-5′ of the arabinofuranose. The correlations from
H-1′′ (δH 4.40) to C-12 (76.8) indicated that the quinovopyranose
The HR‑ESI‑MS of 3 exhibited the molecular formula C36H60O9.
The NMR spectrum of 3 showed similarities to those of 1 and re-
lated compounds in the literature [6], except that the NMR sig-
nals due to the quinovopyranosyl of 1 were replaced by signals
"
attributable to an arabinopyranosyl unit (l Table 2). This sug-
gested that 3 was a triterpenoid arabinopyranoside. In the HMBC
spectrum of 3, long-range heteronuclear correlations from δH H-
"
1
′ (4.32) to δC C-12 (74.8) indicated that the arabinopyranosyl
unit was linked at C-12 (l Fig. 3). The stereochemistry at C-20
Li S et al. New Triterpenoids from… Planta Med 2012; 78: 290–296

2
92 Letters
Table 1 NMR spectroscopic data for cyclocarioside D (1) and cyclocarioside E (2).
Position
Cyclocarioside D (1)
Cyclocarioside E (2)
δC, type
δH (J in Hz)
δC
δH (J in Hz)
1
2
37. 9, CH2
2.49, dt (5.0, 13.2)
37.9
2.49, dt (5.4, 13.2)
1.50, dt (3.0, 13.2)
2.22, dt (3.0, 13.2)
2.69, dt (5.4, 13.2)
1
.51, m
2.22, dt (3.0, 13.2)
.66, dt (5.0, 13.2)
29.8 ca, CH2
30.4 ca
2
3
4
5
6
177.1, C
149.0, C
52.3, CH
25.7, CH2
175.6
149. 0
52.3
2.07, dd (12.9, 2.7)
1.87, m
2.08, dd (12.6, 2.4)
1.87, m
25.7
1
.36, m
1.59, dt (3.6, 13.2)
.18 overlapping
1.36, m
7
35.3, CH2
35.3
1.58, dt (3.3, 13.2)
1.18 overlapping
1
8
9
41.4, C
41.4
40.8
40.4
33.8
40.8, CH
40.4, C
1.89, m
1.88, m
1
1
0
1
33.6, CH2
2.44, td (4.2, 13.2)
2.44, td (4.2, 12.6)
1.40, m
1.40, m
1
1
1
1
2
3
4
5
75.5, CH
44.8, CH
51.3, C
4.13, dt (4.8, 10.8)
1.94 br, d (10.8)
75.6
44.8
51.3
31.6
4.12, dt (4.8, 10.8)
1.94 br, d (10.8)
31.6, CH2
1.06 overlapping
1.06 overlapping
1.42, m
1
.42, m
1.72, m
.22 overlapping
1
6
25.5, CH2
25.5
1.72, m
1
1.22, overlapping
1.81, dt (6.0, 10.2)
1.05, s
1
1
1
2
2
2
2
2
2
2
2
2
7
8
9
0
1
2
3
4
5
6
7
8
50.2, CH
16.8, CH3
20.2, CH3
74.6, C
1.80, dt (4.8, 9.6)
1.06, s
50.2
16.8
20.2
74.6
27.1
45.1
127.3
138.9
75.1
26.5
26.1
113.8
1.11, s
1.10, s
27.1, CH3
45.1, CH2
127.3, CH
138.9, CH
75.1, C
1.17 br, s
1.17 br, s
2.24 br, d (7.8)
5.69, dt (16.0, 7.8)
5.44, d (16.0)
2.23 br, d (7.8)
5.69, dt (16.0, 7.8)
5.44, d (16.0)
26.5, CH3
26.1, CH3
113.7, CH2
1.20, s
1.20, s
1.21, s
1.21, s
4.83 br, s
4.83 br, s
4.71, d (1.8)
1.76 br, s
0.96, s
4.72, d (1.2)
2
3
9
0
1
2
3
4
5
6
24.0 ca, CH3
16.8, CH3
99.8, CH
75.0, CH
77.6, CH
76.7, CH
72.5, CH
18.2, CH3
50.2, CH3
1.76 br, s
24.1 ca
16.7
99.9
75.1
77.6
76.7
72.5
18.2
50.2
60.4
14.6
0.97, s
′
′
′
′
′
′
4.40, d (7.2)
3.21, dd (7.2, 9.0)
3.30, t (9.0)
3.01, t (9.0)
3.27, dd (6.6, 9.0)
1.23, d (6.6)
3.08, s
4.40, d (7.2)
3.25, m
3.30, m
3.01, m
3.27, m
1.23, d (6.0)
3.09, s
OCH3
OCH2CH3
OCH2CH3
4.02, q (7.2)
1.20, t (7.2)
Fig. 2 HMBC (H→C) correlations of compounds 1
and 6.
Li S et al. New Triterpenoids from… Planta Med 2012; 78: 290–296

Letters 293
Table 2 NMR spectroscopic data for cyclocarioside F (3) and cyclocarioside G (4).
Position
Cyclocarioside F (3)
Cyclocarioside G (4)
δC δH (J in Hz)
38.2
δC, type
δH (J in Hz)
1
2
38.4, CH2
2.50, dt (4.8, 13.2)
2.50, dt (4.8, 13.2)
1.50, dt (3.6, 13.2)
2.24, m
1
.50, dt (3.6, 13.2)
2.24, m
.68, ddd (12.6, 5.4, 4.8)
30.4, CH2
30.7
2
2.68, ddd (12.6, 5.4, 4.8)
a
3
4
5
6
177.2 , C
176.1
148.9
52.2
149.0, C
52.3, CH
25.8, CH2
2.08, dd (11.7, 2.7)
1.88, m
2.08, dd (11.7, 2.7)
1.88, m
25.7
1
.33, m
1.60, dt (3.3, 13.2)
.20 overlapping
1.33, m
7
35.4, CH2
35.3
1.60, dt (3.3, 13.2)
1.20 overlapping
1
8
9
41.4, C
41.4
40.8
40.4
33.2
40.8, CH
40.4, C
1.88, m
2.48, m
1.88, m
1
1
0
1
33.0, CH2
2.48, m
1.34, m
1.34, m
1
1
1
1
2
3
4
5
74.8, CH
44.8, CH
51.3, C
4.16, dt (4.8, 11.0)
1.94 br, d (11.0)
75.0
44.8
51.3
31.6
4.16, ddd (4.8, 10.8, 10.8)
1.94 br, d (11.0)
31.6, CH2
1.08 overlapping
1.08, overlapping
1.43, dt (10.8, 9.6)
1.70, m
1
.43, dt (10.8, 9.6)
1.72, m
.22 overlapping
1
6
25.5, CH2
25.5
1
1.22, overlapping
1.81, dt (6.3, 10.5)
1.06, s
1
1
1
2
2
2
2
2
2
2
2
2
7
8
9
0
1
2
3
4
5
6
7
8
50.1, CH
16.9, CH3
20.5, CH3
74.4, C
1.81, dt (6.3, 10.5)
1.06, s
50.2
16.9
20.4
74.5
27.1
45.1
127.2
138.9
75.1
26.5
26.1
113.8
1.10, s
1.10, s
27.1, CH3
45.0, CH2
127.3, CH
138.9, CH
75.1, C
1.18 br, s
1.18 br, s
2.24 br, d (7.8)
5.70, dt (16.0, 7.8)
5.45, d (16.0)
2.24 br, d (7.8)
5.70, dt (16.0, 7.8)
5.45, d (16.0)
26.5, CH3
26.1, CH3
113.8, CH2
1.21, s
1.21, s
1.22, s
1.22, s
4.85 br, s
4.85 br, s
4.73, d (1.8)
1.78 br, s
0.97, s
4.73, d (1.8)
2
3
9
0
1
2
3
4
5
24.0 ca, CH3
16.8, CH3
100.0, CH
72.4, CH
1.78 br, s
24.0 ca
16.8
0.97, s
′
′
′
′
′
4.32, d (7.2)
3.54, m
100.2
72.6
4.32, d (7.2)
3.54, m
73.8, CH
3.52, m
74.0
3.52, m
69.5, CH
3.77 br, s
69.6
3.77, m
67.2, CH2
3.89, dd (10.2, 2.4)
67.2
3.89, dd (10.2, 2.4)
3.54, m
3.54, m
OCH3
50.2, CH3
3.09, s
50.1
60.6
14.5
3.09, s
OCH2CH3
OCH2CH3
4.06, q (6.9)
1.22, t (6.9)
a
Determined from HMBC experiment
and C-24 of 5 were determined to be S and R, respectively, on the
The HR‑EI‑MS of 6 exhibited the molecular formula C31H50O4.
The NMR spectrum of 6 showed a triterpenoid skeleton and a
methoxy group. The seven degrees of unsaturation of 6 con-
firmed that it possessed a 3,4 secodammarane triterpene skele-
1
3
basis of comparison to the C NMR chemical shift data (δC 87.0
and 84.4) of analogous expoxydammaranes, cyclocarioside I [4]
and cyclocarioside A [8], B, and C [7]. In order to determine the
stereochemistry of C-3 and C-12, a NOESY-experiment with a
mixing time of 1.301 sec was carried out. The correlations be-
tween H-3 (3.26) and H-29 (0.88) as well as between H-12
"
ton. The NMR signals of protons and protonated carbons (l Table
3) were assigned by the HSQC and HMBC spectrum correlations
"
(l Fig. 2), together with resolvable homonuclear vicinal coupling
(
4.04) and H-30 (0.94) suggested that H-3 was present in β-orien-
correlations. Long-range heteronuclear correlations from δH 3.64
(-OCH ) to δ 174.4 (C-3) proved that the carboxyl of C-3 was an
tation and H-12 in α-orientation. Therefore, the structure of 5
was deduced as (20S,24R)-epoxydammarane (3α,12β)-25-hy-
droxyl-12-O-β-D-quinovopyranosyl-3-O-(5′-O-acetyl)-α-L-arab-
inofuranoside, and 5 was named cyclocarioside H.
3
C
acetate methyl ester in 6. The stereochemistries at C-12 and C-20
of 6 were determined to be R and S, respectively, on the basis of
1
3
comparison to the C NMR chemical shift data for those corre-
Li S et al. New Triterpenoids from… Planta Med 2012; 78: 290–296

2
94 Letters
Table 3 NMR spectroscopic data for cyclocarioside H (5) and cyclocarin A (6).
Position
Cyclocarioside H (5)
Cyclocarin A (6)
δC, type
δH (J in Hz)
δC, type
δH (J in Hz)
1
2
35.7, CH2
2.50, dt (13.8, 3.6)
34.3, CH2
1.63, m
1
.30, m
1.54, m
.70, m
21.2, CH2
28.4, CH2
2.18, m
2.33, m
1
3
4
5
6
79.6, CH
38.1, C
3.26, t (2.4)
174.4, C
147.2, C
50.6, CH
24.6, CH2
51.2, CH
18.6, CH2
1.28, m
1.42, m
1.97, br, dd (12.5, 2.5)
1.38, m
1
.79, t (10.5)
1.50, m
.24, br, d (13.0)
7
36.8, CH2
1.25, m
33.4, CH2
1
8
9
41.9, C
39.0, C
54.5, CH
40.2, C
1.68, m
40.5, CH
39.3, C
1.63, m
1
1
0
1
34.6, CH2
2.46, dt (12.6, 4.8)
30.9, CH2
1.06 br, d (8.8)
1.32, m
1.32, m
1
1
1
1
2
3
4
5
76.8, CH
41.4, CH
50.6, C
4.04, m
1.70, m
70.6, CH
48.2, CH
52.1, C
3.60, m
1.79, t (10.5)
32.0, CH2
1.06, m
31.6, CH2
1.38, m
1.94, m
1.87, m
1
.42, m
1.43, m
.84, m
1
6
27.1, CH2
26.4, CH2
1
17
18
19
20
21
22
49.6, CH
17.0, CH3
16.7, CH3
87.0, C
1.84, m
1.00, s
1.10, s
52.5, CH
15.4, CH3
20.1, CH3
74.2, C
2.12, m
1.02, s
0.88, s
24.8, CH3
34.2, CH2
1.12, s
27.6, CH3
39.4, CH2
1.19, s
1.66, m
2.27, dd (13.0, 8.5)
2
.45, dd (13.0, 8.0)
23
24
25
26
26.3, CH2
84.4, CH
71.7, C
1.84, m
125.7, CH
136.5, CH
141.8, C
5.74, dt (16.0, 8.0)
6.22, d (16.0)
3.74, d (7.2)
26.3, CH3
1.16, s
115.2, CH2
4.90, br, s
4
.89, br, s
2
2
7
8
26.9, CH3
1.14, s
0.92, s
18.7, CH3
1.85, s
29.8 ca, CH3
113.6, CH2
4.86, br, s
4
.66, br, s
2
3
1
2
3
4
5
9
0
′
22.9, CH3
17.2, CH3
106.0, CH
83.8, CH
79.8, CH
82.1, CH
65.0, CH2
0.88, s
23.2, CH3
16.7, CH3
1.73, s
0.89, s
0.94, s
4.93, br, s
′
4.08, m
51.6, CH3
3.64, s
′
3.84, dd (6.3, 6.3)
4.08, m
′
′
4.26, dd (11.4, 3.0)
4.10, dd (11.4, 6.6)
CH3COO
CH3COO
170.8, C
20.7, CH3
100.8, CH
75.3, CH
78.0, CH
76.8, CH
72.4, CH
18.2, CH3
2.00, s
1
2
3
4
5
6
′′
′′
′′
′′
′′
′′
4.40, d (7.8)
3.10, dd (9.0, 7.8)
3.31, dd (9.0, 9.0)
2.99, dd (9.0, 9.0)
3.28, dd (9.0, 6.0)
1.22, d (6.0)
sponding to 3,4-secodammaranes compounds [19]. Therefore,
the structure of 6 was deduced as (23E)(12R,20S)-12,20-dihy-
droxy-3,4-secodammarane-4(28),23,25-trien-3-oic acid methyl
ester, and 6 was named cyclocarin A.
Compounds 1–6 were evaluated against several human cancer
cell lines and were not active (IC50>10 µM). They were screened
for inhibitory activities against α-glucosidase (40 µM), lipase
(5 µM), DPP‑IV (10 µM), and aldose reductase (10 µM), respective-
ly. These compounds showed weak activities at the same concen-
tration as the positive control drugs (acarbose, orlistat, INDP-2,
"
and epalrestat, see l Table 4).
Li S et al. New Triterpenoids from… Planta Med 2012; 78: 290–296

Letters 295
Table 4 Assessment of α-glucosidase, lipase, DPP‑IV, and aldose reductase in-
hibitory activity (%) of compounds 1–6.
Com-
α-Gluco-
Lipase
DPP‑IV
Aldose
pound
sidase
reductase
1
11.4
10.7
8.6
8.9
14.7
9.5
11.1
11.4
23.2
11.1
17.1
15.7
9.5
6.8
2
3
5.3
4
3.6
3.7
10.0
9.1
5
1.5
5.7
6
2.9
8.4
9.1
Acarbose
Orlistat
INDP-2
Epalrestat
100
100
97.50
100
Isolates: Cyclocarioside D (1): colorless needles (MeOH), m.p.
2
0
1
36.5–139.5°C; R = ca. 0.55 (CHCl -MeOH‑HAc, 5:1:0.1); [α]
D
f
3
"
Fig. 3 Main HMBC (H→C) correlations of compound 5.
+ 4.6 (c 0.09, MeOH); NMR data, see l Table 1; FAB‑MS m/z (%):
73 [M + Na] (12); HR‑ESI‑MS m/z 673.4288 [M + Na] (calcd.
+
+
6
for C37H62O Na, 673.4286).
9
Cyclocarioside E (2): colorless powder (MeOH), m.p. 127.5–
2
0
Materials and Methods
129.6°C; R = ca. 0.63 (CHCl -MeOH, 10:1); [α]
D
+ 4.8 (c 0.10,
f
3
MeOH). NMR data, see l
Table 1; FAB‑MS m/z (%): 701 [M + Na]+
(13); HR‑ESI‑MS 701.4616 [M + Na]+ (calcd. for C39H66O Na
"
!
The leaves of C. paliurus were collected at Qimen, Anhui Province,
China, in September 2001, and were identified by Mr. Ma-Lin (In-
stitute of Materia Medica, Chinese Academy of Medical Sciences).
A voucher specimen (No. ZH02001) has been deposited at the
Herbarium of the Department of Medicinal Plants, Institute of
Materia Medica, Chinese Academy of Medical Sciences.
9
701.4599).
CyclocariosideF(3):colorlesspowder(MeOH),m.p. 133.0–136.5°C;
+ 12.4 (c 0.10, MeOH).
NMR data, see l Table 2; FAB‑MS m/z (%): 659 [M + Na] (4);
2
0
R = ca. 0.34 (CHCl -MeOH, 10:1); [α]
D
f
3
"
+
+
HR‑ESI‑MS 659.4133 [M + Na] (calcd. for C36H60O Na 659.4135).
9
The air-dried and powdered leaves of C. paliurus (1 kg) were
extracted with 95% ethanol (6 L × 1 h × 3) by ultrasonication
Cyclocarioside G (4): colorless powder (MeOH), m.p. 114.5–
20
116.8°C; R = ca. 0.60 (CHCl -MeOH, 10:1); [α]
MeOH); NMR data, see l Table 2; FAB‑MS m/z (%): 687[M + Na]
(3); HR‑FAB‑MS 687.4425 [M + Na]+ (calcd. for C38H64O Na
D
+ 2.8 (c 0.11,
f
3
"
+
(230 W, 35 KHz) at room temperature. The extracts were com-
bined and concentrated under reduced pressure (at < 60°C) to
give a dark brown residue (51 g). The residue was suspended in
water (3 L) and then partitioned with ethyl acetate (3 L × 4, 29 g).
The ethyl acetate fraction (28 g) was subjected to column chro-
matography (CC) [silica gel (mesh 45–75 µm; 500 g), column,
9
687.4448).
Cyclocarioside H (5): was obtained as colorless powder (MeOH),
m.p. 129.5–133.4°C; R = ca. 0.54 (CHCl -MeOH‑HAc, 5:1:0.1);
f
3
20
"
[α]
D
− 31.4 (c 0.11, MeOH); NMR data, see l Table 3; FAB‑MS m/z
(%): 797 [M + H] (0.7); HR‑ESI‑MS 819.4867 [M + Na] (calcd. for
C43H72O13Na 819.4865).
+
+
6
× 70 cm] eluted with a gradient of increasing methanol (0–20%)
in chloroform and methanol to yield fourteen fractions (A–N) on
the basis of TLC analyses. Fraction J [CHCl -MeOH (100:1, 6.0 L),
Cyclocarin A (6): colorless powder (MeOH), m.p. 129.2–130.2°C;
3
20
2.3 g] was applied to CC [silica gel (mesh 45–75 µm; 60 g), col-
R = ca. 0.43 (CHCl -MeOH, 20:1); [α]
NMR data, see l Table 3; EI‑MS m/z (%): 486 (0.4), 468 (2.3), 405
(22), 387 (100), 355 (35), 85 (22), 71 (29), 57 (42); HR‑EI‑MS m/z
D
+ 18.6 (c 0.06, MeOH).
f
3
"
umn, 2 × 45 cm] eluted with CHCl -MeOH (40:1, 3.5 L) to give
3
the subfractions J2 (651 mg) and J3 (839 mg). The subfraction J2
was purified by CC [Sephadex LH-20 (50 g), column, 1.5 × 60 cm]
486.3702 (calcd. for C31H50O 486.3709).
4
eluted with CHCl -MeOH and was applied to preparative HPLC
Acid hydrolysis: Solutions of 1, 2, 3, 4, or 5 were mixed with 0.5 N
HCl and refluxed for 1 h. Each reaction mixture was diluted with
water and extracted exhaustively with CHCl . The CHCl layers
3
[
column, RP-18 (250 × 10 mm, 5 µm); MeOH‑H O (70:30), 2.0 L,
2
2
mL/min flow rate] to obtain 2 (35 mg) and 4 (44 mg). Fraction
3
3
K (0.9 g) was purified by CC [Sephadex LH-20 (50 g), column,
were dried over anhydrous Na SO4 and evaporated under re-
2
1
.5 × 60 cm] eluted with petroleum ether-CHCl -MeOH (5:5:1,
duced pressure. Examination of each CHCl layer by TLC showed
3
3
4
50 mL) to give the subfraction K2 (690 mg) which was applied
a number of products which were less polar than the starting ma-
terials. The aqueous layer was neutralized with sodium bicarbon-
ate and allowed to dry at room temperature. TLC analysis of the
residues using CHCl -MeOH‑H O (6:4:1) showed the presence
to CC [silica gel (mesh 45–75 µm; 30 g), column, 1.5 × 30 cm]
eluted with CHCl -MeOH (50:1, 1.9 L) to yield 6 (12 mg). Fraction
L (2.5 g) was purified by CC [Sephadex LH-20 (50 g), column,
3
3
2
1
.5 × 60 cm] eluted with petroleum ether-CHCl -MeOH (5:5:1,
of quinovose or arabinose as the only sugars for 1, 2, 3, and 4,
and both quinovose and arabinose for 5.
Assessment of several human cancer cell lines inhibitory activity:
See previous report [21].
3
1.6 L) to give six subfractions (L -L ). The L subfraction (1.2 g)
1
6
3
was subjected to CC [silica gel (mesh 45–75 µm; 50 g), column,
× 45 cm] eluted with CHCl -MeOH (20:1, 3.0 L) and then ap-
2
3
plied to preparative HPLC [column, RP-18 (250 × 10 mm, 5 µm);
Assessment of DPP‑IV inhibitory activity: See previous report
[22].
MeOH‑H O (70:30), 2.0 L, 2 mL/min flow rate] to obtain 1
2
(30 mg), 3 (80 mg), and 5 (221 mg).
Li S et al. New Triterpenoids from… Planta Med 2012; 78: 290–296

2
96 Letters
1
1
0 Zhong RJ, Gao YH, Xu CR, Li LN. Pentacyclic triterpenoids from round-
uingfruit Cyclocarya (Cyclocarya paliurus). Chin Tradit Herb Drugs
Supporting information
UV, IR, MS, 1H NMR, and 13C NMR spectra as well as 2D NMR cor-
1
996; 27: 387–388
relation spectra of compounds 1–6, the key chemical method of
sugar moieties acid hydrolysis, and TLC identification are avail-
able as Supporting Information.
The detailed protocols for assaying α-glucosidase, lipase, and al-
dose reductase inhibition are also available as Supporting Infor-
mation.
1 Shu RG, Song ZR, Shu JC. Study on the chemical constituents of the bu-
tanol extraction of Cyclocarya paliurus (Batal.) Iljinsk. J Chin Med Mater
2006; 29: 1304–1307
2 Shu RG, Shu JC. Phenolic constituents of Cyclocarya paliurus. Chin Tradit
Herb Drugs 2007; 38: 507–508
1
1
1
3 Li J, Lu YY, Li F, Zhang YJ, Huang XS, Su XJ. Study on chemical constitu-
ents of Cyclocarya paliurus. J Chin Med Mater 2006; 29: 441–442
4 Li J, Lu YY, Xu ZJ, Li F, Wen GQ, Su XJ. Studies on flavones in the leaves of
Cyclocarya paliurus (Batal.) Iljinsk. J Chin Med Mater 2005; 28: 1058–
Acknowledgements
!
1
059
1
5 Zhang XQ, Ye WC, Yin ZQ, Zhang ZH, Zhao SX. Study on the chemical con-
stituents of Cyclocarya paliurus. Chin J Chin Mater Med 2005; 30: 791–
Thanks are due to Prof. Xiaoguang Chen for the human cancer
cells screening and Prof. Jiangong Shi for guidance. Thanks are
also due to Dr. Alison Rinderspacher for revision of the manu-
script. Financial support by the National Science and Technology
Project of China (No. 2009ZX09301-003-4-1) for part of this work
is acknowledged.
7
92
16 Zhang J, Lu JC, Xiao K, Lü T, Fan JW, Zhang LM, Zhao J, Yang TJ. Studies on
water-soluble constituents in the leaves of Cyclocarya paliurus. J Pharm
Pract 2007; 25: 82–84
1
7 Yi X, Shi JG, Zhou GX, Xie MY. Studies on the chemical constituents in the
leaves of Cyclocarya paliurus. Chin J Chin Mater Med 2002; 27: 43–45
8 Shu RG, Liu YF, Chen J, Shu JC. Studies on the triterpenoids of Cyclocarya
paliurus (Batal.) Iljinsk. J Chin Med Mater 2005; 28: 558–559
1
Conflict of Interest
!
19 Tadashi A, Shinji O, Takayuki S. Six novel secodammarane-tape triter-
penes from male flower of Alnus japonica. Phytochemistry 1988; 27:
The corresponding authors declare that the manuscript is sub-
mitted on behalf of all authors. There are no conflicts of interest
among all authors of the manuscript. Copyright belongs to the
publisher upon acceptance of the manuscript.
2
915–2920
2
0 Dewick PM. Medicinal natural products – a biosynthetic approach. Chi-
chester: John Wiley & Sons, Ltd.; 2002
21 Xue Z, Li S, Wang SJ, Wang YH. Mono-, bi-, and triphenanthrenes from
the tubers of Cremastra appendiculata. J Nat Prod 2006; 69: 907–913
2
2 Hu CX, Huang H, Zhang L, Huang Y, Shen ZF, Cheng KD, Du GH, Zhu P.
A new screening method based on yeast-expressed human dipeptidyl
peptidase IV and discovery of novel inhibitors. Biotechnol Lett 2009;
References
1
Editor Committee for Flora of China of the Chinese Academy of Science.
Flora of China, Volume 21. Beijing: Science Publishing House; 1979:
3
1: 979–984
1
8–19
2
The Editorial Committee of the Administration Bureau of Traditional Chi-
nese Medicine. Chinese materia medica (Zhonghua Bencao), Volume 2.
Shanghai: Shanghai Science and Technology Publishing House; 1999:
received July 10, 2011
revised
November 11, 2011
3
70–371
accepted November 14, 2011
3
Kurihara H, Asami S, Shibata H, Fukami H, Tanaka T. Hypolipemic effect
of Cyclocarya paliurus (Batal) Iljinskaja in lipid-loaded mice. Biol Pharm
Bull 2003; 26: 383–385
Bibliography
DOI http://dx.doi.org/10.1055/s-0031-1280411
Published online December 7, 2011
Planta Med 2012; 78: 290–296
4
5
Shu RG, Xu CR, Li LN. Studies on the sweet principles from the leaves of
Cyclocarya paliurus (Batal.) Iljinsk. Acta Pharm Sin 1995; 30: 757–761
Shu RG, Xu CR, Li LN, Yu ZL. Cyclocariosides II and III: two secodammar-
ane triterpenoid saponins from Cyclocarya paliurus. Planta Med 1995;
©
Georg Thieme Verlag KG Stuttgart · New York ·
ISSN 0032‑0943
6
1: 551–553
6
Kennelly EJ, Cai LN, Long LN, Shamon L, Zaw K, Zhou BN, Pezzuto JM,
Kinghorn AD. Novel highly sweet secodammarane glycosides from
Pterocarya puliurus. J Agric Food Chem 1995; 43: 2602–2607
Jiang ZY, Zhang XM, Zhou J, Qiu SX, Chen JJ. Two new triterpenoid glyco-
sides from Cyclocarya paliurus. J Asian Nat Prod Res 2006; 8: 93–98
Yang DJ, Zhong ZC, Xie ZM. Studies on the sweet principles from the
leaves of Cyclocarya paliurus (Batal.) Iljinskaya. Acta Pharm Sin 1992;
Correspondence
Dr. Shuai Li
Institute of Materia Medica, Department of Natural Product Chemistry
Chinese Academy of Medical Sciences and Peking Union Medical College
1 Xian Nong Tan Street
Beijing 100050
P.R. China
7
8
2
7: 841–844
Phone: + 861063164628
Fax: + 861063017757
lishuai@imm.ac.cn
9
Zhong RJ, Shu RG, Ni XL, Xu CR, Li LN. Studies on the chemical structure
of cyclocaric acid A. Acta Pharm Sin 1996; 31: 398–400
Li S et al. New Triterpenoids from… Planta Med 2012; 78: 290–296

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