Neolignans and caffeoyl derivatives from Selaginella moellendorffii

Article abstract of DOI:10.1002/hlca.201000116

Ten new phenolic compounds including the six neolignans 1-3 and 6-8 and four caffeoyl derivatives, i.e., myo-inositol 1-caffeate (9), myo-inositol 6-caffeate (10), myo-inositol 5-caffeate (11), and paucine 3′-β-D- glucopyranoside (12) were isolated from the whole plants of Selaginella moellendorffii (caffeic acid=3-(3,4-dihydroxyphenyl)prop-2-enoic acid). Their structures were established by spectroscopic and chemical methods.

Full text of DOI:10.1002/hlca.201000116

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Neolignans and Caffeoyl Derivatives from Selaginella moellendorffii
by Yue-Hu Wanga), Qian-Yun Sunb), Fu-Mei Yangb), Chun-Lin Long*a)c), Fu-Wei Zhaoa), Gui-Hua
Tanga), Hong-Mei Niua)d), Huan Wanga)e), Qiao-Qin Huanga)e), Jin-Jin Xua)e), and Li-Juan Maa)c)
a) Key Laboratory of Economic Plants and Biotechnology, Kunming Institute of Botany,
Chinese Academy of Sciences, Kunming 650204, P. R. China
(phone/fax: þ 86-10-6893 0381; e-mail: long@mail.kib.ac.cn)
b) Key Laboratory of Chemistry for Natural Products, Guizhou Province and Chinese Academy of
Sciences, Guiyang 550002, P. R. China
c) College of Life and Environmental Sciences, Minzu University of China, Beijing 100081, P. R. China
d) Southwest Forestry University, Kunming 650224, P. R. China
e) Yunnan Agricultural University, Kunming 650201, P. R. China
Ten new phenolic compounds including the six neolignans 1 – 3 and 6 – 8 and four caffeoyl
derivatives, i.e., myo-inositol 1-caffeate (9), myo-inositol 6-caffeate (10), myo-inositol 5-caffeate (11),
and paucine 3’-b-d-glucopyranoside (12) were isolated from the whole plants of Selaginella moellen-
dorffii (caffeic acid ¼ 3-(3,4-dihydroxyphenyl)prop-2-enoic acid). Their structures were established by
spectroscopic and chemical methods.
Introduction. – Selaginella moellendorffii Hieron. (Selaginellaceae), a perennial
herb, is mainly distributed in the southern area of the Changjiang River in China and
has been used to treat jaundice, gonorrhea, bleeding, acute hepatitis, and idiopathic
thrombocytopenic purpura (ITP) in traditional Chinese folk medicine. Earlier studies
found a coumarin, fatty acids [1], a lignan glucoside [2], flavone glycosides [3], and
cytotoxic [4] and antioxidative [5] biflavones in this plant. In our recent research, a
series of pyrrolidinoindoline alkaloids were found from this plant [6]. In continuation
of the studies on this plant, ten new phenolic derivatives, 1 – 3 and 6 – 12, and four
known ones, compounds 4 and 5, paucine (¼(2E)-N-(4-aminobutyl)-3-(3,4-dihydroxy-
phenyl)prop-2-enamide 13) [7], and N1-cis-p-coumaroylagmatine (¼(2Z)-N-{4-[(ami-
noiminomethyl)amino]butyl}-3-(4-hydroxyphenyl)prop-2-enamide 14) [8], were iso-
lated from the whole plants of S. moellendorffii. The structure elucidation of these new
compounds is reported.
Results and Discussion. – Compound 11) was obtained as an amorphous solid with
an m/z of 441.1518 ([M þ Na]þ) by HR-ESI-MS, leading us to assign it a molecular
formula C22H26O8. The IR spectrum of 1 showed absorptions for hydroxy (3441 cmꢀ1),
carbonyl (1731 cmꢀ1), and aromatic groups (1616, 1519, and 1500 cmꢀ1). In the
13C-NMR spectrum of 1 (Table 1), 22 signals for 4 MeO groups (d(C) 56.1 (2q), 55.8
(q), and 51.5 (q)), 12 aromatic (d(C) 147.1 (2s), 146.4 (s), 143.9 (s), 134.5 (s), 133.7 (s),
1
)
Trivial atom numbering; for systematic names, see Exper. Part.
ꢀ 2010 Verlag Helvetica Chimica Acta AG, Zꢁrich

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132.2 (s), 128.2 (s), 116.1 (d), 112.1 (d), and 103.0 (2d)), 1 ester carbonyl (d(C) 173.7
(s)), and 5 alphatic C-atoms (d(C) 88.0 (d), 63.5 (t), 53.7 (d), 36.0 (t), and 30.6 (t))
suggested that this compound might be a lignan. Careful comparison of the NMR data
of 1 (Table 1) with those of known neolignans revealed that compound 1 is a
dihydrobenzofuran neolignan with oxidation of C(9’) and methoxy substituentes at
C(3), C(3’), C(5), and C(9’) [9][10], which was confirmed by the HMBCs of
MeOꢀC(3,5)/C(3,5), MeO-C(3’)/C(3’), and MeO-C(9’)/C(9’) (Fig.). The relative
Figure. Key HMBCs (H ! C) of 1

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configuration of HꢀC(7) and HꢀC(8) was assigned as trans by the ROESY
correlations of HꢀC(7)/CH2(9); J values are useless to distinguish cis/trans diaster-
eoisomers of the dihydrobenzofuran neolignans [9]. On the other hand, a significant
difference in chemical shifts of C(8) in 9-hydroxydihydrobenzofuran neolignans,
appearing at d ca. 54 in the trans isomers and at d ca. 49 in the cis ones [9][11], is a
reliable criterion to distinguish both diastereoisomers. The chemical shift of C(8) (d
53.7) in 1 also suggested the relative trans-configuration of HꢀC(7) and HꢀC(8).
The absolute configurations of dihydrobenzofuran neolignans are usually deter-
mined by the signs of the band 1Lb (270 – 300 nm) or 1La (220 – 240 nm) in the circular-
1
dichroism (CD) spectrum. The positive signs of the Lb band predict the absolute
configuration of 7,8-trans-3-methoxydihydrobenzofuran neolignans to be (7S,8R)
[12][13]. The CD spectrum of 1 showed a positive Cotton effect at 282 nm (De þ 0.13),
indicating that the absolute configuration of 1 was the (7S,8R)-configuration.
Therefore, compound 1 was elucidated as (7S,8R)-4,9-dihydroxy-3,3’,5-trimethoxy-
4’,7-epoxy-8,5’-neolignan-9’-oic acid methyl ester1).
Compound 21) was obtained as colorless amorphous solid. The HR-ESI-MS of 2
had a molecular-ion peak at m/z 401.1236 ([M ꢀ H]ꢀ), consistent with the molecular
formula C21H22O8. The 13C-NMR spectra of 2 (Table 1) were very similar to those of 1,
except for the signals of the side chain at 1 at d(C) 30.6 (t, C(7’)), 36.0 (t, C(8’)), 173.7 (s,
C(9’)), and 51.5 (q, MeOꢀC(9’)) which were replaced by those of 2 at d(C) 146.8 (d,
C(7’)), 116.4 (d, C(8’)), and 170.9 (s, C(9’)), implying that C(7’) and C(8’) were
dehydrogenated and the carboxyl group (C(9’)) was not esterified in 2. The
C(7’)¼C(8’) bond had (E)-configuration according to the coupling constant
(J(7’,8’) ¼ 16.0 Hz, Table 1), and the relative configuration of 2 was determined as
trans by the chemical shift of C(8) (d(C) 55.0). The structure of 2 was thus confirmed to
be rel-(7R,7’E,8S)-4,9-dihydroxy-3,3’,5-trimethoxy-4’,7-epoxy-8,5’-neolign-7’-en-9’-oic
acid1).
Compound 31) was assigned the molecular formula C16H14O5 by HR-ESI-MS
analysis (m/z 285.0754 ([M ꢀ H]ꢀ, C16H13Oꢀ5 )). The IR spectrum showed absorption
bands at 3397, 1674, 1611, 1600, and 1518 cmꢀ1 due to hydroxy, carbonyl, and phenyl
groups. The 1H-NMR spectrum (Table 1) of 3 indicated the presence of a p-
disubstituted (d(H) 7.18 (d, J ¼ 8.5 Hz, 2 H) and 6.76 (d, J ¼ 8.5 Hz, 2 H)) and a 1,2,4-
trisubstituted (d(H) 7.94 (br. s), 7.91 (dd, J ¼ 8.5, 1.5 Hz), and 6.84 (d, J ¼ 8.5 Hz))
benzene ring in 3. Its 13C-NMR spectrum (Table 1) showed 16 signals, including the
characteristic ones for dihydrobenzofuran neolignans (d(C) 89.6 (d), 64.7 (t), and 54.1
(d)), 12 ones for two benzene rings (d(C) 165.4, 158.6, 133.6, 133.0, 129.7, 128.3 (2 C),
128.1, 124.4, 116.4 (2 C), and 109.9), and one for a carboxy group (d(C) 170.0), which
implied that this compound might be a 8’,9’-dinor-dihydrobenzofuran neolignan. This
was confirmed by the HMBC spectrum of 3. The relative configuration of 3 was also
determined as trans by the chemical shift of C(8) (d(C) 54.1), and the absolute
configuration of 3 was assigned as (7S,8R) by comparing its CD data with those of the
known (þ)-conocarpan [14 – 16]. Therefore, compound 3 was determined as (7S,8R)-
4,9-dihydroxy-4’,7-epoxy-8’,9’-dinor-8,5’-neolignan-7’-oic acid1).
Compound 41) showed the [M þ Cl]ꢀ ion peak at m/z 425.1370 in the negative-ion-
mode HR-ESI-MS, indicating the molecular formula C21H26O7. The NMR spectra of 4
(Table 1) were very similar to those of (þ)-(7R,8S)-5-methoxydihydrodehydroconi-

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feryl alcohol [10]. However, the optical rotation value of 4 was negative rather than
positive, implying the absolute configuration of 4 to be (7S,8R), which is consistent
with those of 1 and 3. Consequently, 4 is (7S,8R)-3,3’,5-trimethoxy-4’,7-epoxy-8,5’-
neolignan-4,9,9’-triol1). An analog without optical activity was also reported in two
other references [17][18].
The molecular formula of 51) was determined as C27H36O12 by the [M þ Cl]ꢀ ion
peak at m/z 587.1883 in the HR-ESI-MS. The NMR data of 5 (Table 2) were very
similar to those of a known neolignan glucoside [19]. The relative configuration of
HꢀC(7) and HꢀC(8) in the known compound was suggested to be cis according to the
J value between the two protons by the authors. It should be revised as trans based on
the chemical shift of C(8) (d(C) 55.8) [9][11]. Because the optical value of this
compound has not been reported, it was difficult to determine if compound 5 is
identical to this known one. Acidic hydrolysis of 5 provided a d-glucose. Thus, 5 was
deduced as rel-(7R,8S)-3,3’,5-trimethoxy-4’,7-epoxy-8,5’-neolignan-4,9,9’-triol 4-b-d-
glucopyranoside1).
Compound 61) had the molecular formula C27H36O12 as deduced from an HR
negative-ion-mode ESI-MS. Its NMR data (Table 2) showed signals for a neolignan
moiety identical to that of 4 and a b-glucose moiety (d(H) 4.36 (d, J ¼ 7.9 Hz,
HꢀC(1’’))). The glucosyloxy group was located at C(9) by the long-range correlations
between HꢀC(9) (d(H) 4.10 (t, J ¼ 8.5 Hz)) and C(1’’) (d(C) 104.1), and HꢀC(1’’) to
C(9) (d(C) 72.3) in the HMBC spectrum of 6. Therefore, 6 is rel-(7R,8S)-3,3’,5-
trimethoxy-4’,7-epoxy-8,5’-neolignan-4,9,9’-triol 9-b-d-glucopyranoside1).
Compound 71) was found to have the formula C27H34O12, two H-atoms less than that
of 6, from the HR-ESI-MS (neg.). Comparison of the NMR spectra (Table 2) and the
formula of 7 with those of 6 indicated that 7 is a 7,8-didehydro derivative of 6, namely
3,3’,5-trimethoxy-4’,7-epoxy-8,5’-neolign-7-ene-4,9,9’-triol
which was further confirmed by the HMBC spectrum of 7.
9-b-d-glucopyranoside1),
Compound 81) was shown to have a molecular formula of C28H40O13 by HR-ESI-MS
(neg.). Signals for two 1,2,3,5-tetrasubstituted benzene moieties (d(H) 6.60 (s,
HꢀC(2,6)) and 6.50 (s, HꢀC(2’,6’))) and an anomeric proton (d(H) 4.80 (d, J ¼
7.9 Hz )) of a b-glucosyl group were observed in the 1H-NMR spectrum of 8 (Table 2).
In the 13C-NMR spectrum of 8 (Table 2), 28 signals for 4 MeO (d(C) 57.5 (2q) and 57.0
(2q)), two aromatic (d(C) 154.5 (2s), 153.8 (2s), 139.8 (s), 136.9 (s), 134.8 (s), 134.6 (s),
108.4 (2d), and 106.7 (2d)), and a glucosyl group (d(C) 105.6 (d), 78.3 (d), 77.8 (d), 75.7
(d), 71.2 (d), and 63.6 (t)), and 6 other alphatic C-atoms (d(C) 84.9 (d), 63.6 (t), 62.1
(t), 39.0 (t), 35.4 (t), and 33.4 (t)) implied that this compound might be a lignan
glucoside, which was further determined as a 8,4’-oxyneolignan 4-glucoside by the
HMBC cross-peaks HꢀC(8)/C(4’) and HꢀC(1’’)/C(4). Accordingly, compound 8 was
elucidated as 3,3’,5,5’-tetramethoxy-8,4’-oxyneolignane-4,9,9’-triol 4-b-d-glucopyrano-
side1).
Compound 9 was obtained as a colorless amorphous solid. The negative-ion-mode
HR-ESI-MS exhibited an accurate ion peak at m/z 341.0878 ([M ꢀ H]ꢀ), in accordance
with the molecular formula C15H18O9. The IR spectrum showed absorption bands due
to hydroxy (3407 cmꢀ1), conjugated carbonyl (1688 and 1631 cmꢀ1), and phenyl (1604,
1523, and 1445 cmꢀ1) groups. A caffeoyl (d(H) 7.02 (s), 6.98 (d, J ¼ 8.0 Hz), 6.75 (d, J ¼
8.0 Hz), 7.49 (d, J ¼ 16.0 Hz), and 6.24 (d, J ¼ 16.0 Hz); d(C) 166.5 (s, C(9’))) and a

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myo-inositol group (d(H) 4.50 (br. d, J ¼ 9.6 Hz, HꢀC(1)), 3.84 (br. s, HꢀC(2)), 3.21
(br. d, J ¼ 9.6 Hz, HꢀC(3)), 3.40 (t, J ¼ 9.6 Hz, HꢀC(4)), 3.00 (t, J ¼ 9.6 Hz, HꢀC(5)),
and 3.64 (t, J ¼ 9.6 Hz, HꢀC(6)) were easily deduced from its NMR spectra (Table 3).
The two moieties were linked through the bonds C(1)ꢀOꢀC(9’) by the long-range
correlation HꢀC(1)/C(9’) in the HMBC spectrum of 9. Therefore, compound 9
was elucidated as myo-inositol 1-caffeate. Its absolute configuration was not deter-
mined.
Table 3. 1H- and 13C- NMR Data (400 and 100 MHz, resp.; (D6)DMSO) of 9 – 11. d in ppm, J in Hz.
9
10
11a)
d(H)
d(C) d(H)
d(C) d(H)
d(C)
HꢀC(1) 4.50 (br. d, J ¼ 9.6)
HꢀC(2) 3.84 (br. s)
HꢀC(3) 3.21 (br. d, J ¼ 9.6)
HꢀC(4) 3.40 (t, J ¼ 9.6)
HꢀC(5) 3.00 (t, J ¼ 9.6)
HꢀC(6) 3.64 (t, J ¼ 9.6)
C(1’)
74.6 3.40 (dd, J ¼ 9.6, 2.4)
70.1 3.74 (t, J ¼ 2.4)
71.4 3.19 (dd, J ¼ 9.6, 2.4)
72.4 3.43 (t, J ¼ 9.6)
75.2 3.13 (t, J ¼ 9.6)
70.1 5.03 (t, J ¼ 9.6)
125.5
69.8 3.20 – 3.27 (m)
73.0 3.69 – 3.75 (m)
71.6 3.20 – 3.27 (m)
72.8 3.55 (ddd, J ¼ 10.0, 10.0, 5.5)
75.3 4.64 (t, J ¼ 10.0)
75.3 3.55 (ddd, J ¼ 10.0, 10.0, 5.5)
125.8
115.3 7.03 (s)
145.6
148.2
71.8
72.6
71.8
70.7
76.7
70.7
125.8
114.7
145.6
148.2
115.8
121.2
144.3
115.3
166.2
HꢀC(2’) 7.02 (s)
114.6 7.03 (d, J ¼ 1.7)
C(3’)
145.7
C(4’)
148.6
HꢀC(5’) 6.75 (d, J ¼ 8.0)
HꢀC(6’) 6.98 (d, J ¼ 8.0)
HꢀC(7’) 7.49 (d, J ¼ 16.0)
HꢀC(8’) 6.24 (d, J ¼ 16.0)
C(9’)
115.8 6.74 (d, J ¼ 8.2)
115.8 6.73 (d, J ¼ 8.0)
121.3 6.97 (dd, J ¼ 8.2, 1.7) 121.2 6.97 (d, J ¼ 8.0)
144.9 7.49 (d, J ¼ 15.8)
114.5 6.23 (d, J ¼ 15.8)
166.5
144.3 7.40 (d, J ¼ 16.0)
114.6 6.24 (d, J ¼ 16.0)
166.4
a) 1H-NMR Signals of the OH groups: 9.56 (s, OHꢀC(4’)); 9.18 (s, OHꢀC(3’)); 4.76 (d, J ¼ 5.5, OHꢀC(4),
OHꢀC(6)); 4.66 (overlapped, OHꢀC(2)); 4.60 (d, J ¼ 5.5, OHꢀC(1), OHꢀC(3)).
Compounds 10 and 11 had the same molecular formula C15H18O9 as compound 9, as
determined by HR-ESI-MS (neg.). Based on the MS and NMR spectra (Table 3), 10
and 11 also comprised a caffeoyl and a myo-inositol moiety. The differences concerned
the linkage positions of the two fragments. The caffeoyl group was located at OꢀC(6)
in 10 and at OꢀC(5) in 11 as shown by the HMBC spectra. Thus, compounds 10 and 11
were deduced as myo-inositol 6-caffeate and myo-inositol 5-caffeate, respectively.
Generally, hydroxy cinnamic acids including caffeic acid are esters bound to various
alcohols, most frequently sugars [20]. However, only a few inositol hydroxycinnamates
were reported. As far as known, myo-inositol trans- and cis-p-coumarates were isolated
from two Taxus plants [21][22] and several myo-insitol ferulate derivatives were
synthesized [23]. Compounds 10 and 11 represent the first examples of inositol caffeate
derivatives.
Compound 12 was obtained as a colorless amorphous solid having the molecular
formula C19H28N2O8 as determined by HR-ESI-MS ([M þ H]þ at m/z 413.1933). The
IR spectrum of 12 showed absorptions for hydroxy (3439 cmꢀ1), carbonyl (1639 cmꢀ1),
and aromatic (1630, 1547, and 1511 cmꢀ1) groups. The NMR spectra of 12 indicated the

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presence of a b-glucopyranose, a caffeoyl, and a putrescine (¼ butane-1,4-diamine).
Comparison of the NMR data of 12 with those of paucine (13) revealed that compound
12 is a glucoside of paucine [7]. The glucose was located at C(3’) according to the
HMBC cross-peak HꢀC(1’’)/C(3’). Therefore, compound 12 was elucidated as paucine
3’-b-d-glucopyranoside.
Compounds 7 and 11, and N1-cis-p-coumaroylagmatine (14) were tested for
biological activity against the leukemia K562 cell line, and compound 12 and paucine
against acetylcholinesterase (AChE). However, none of these compounds showed
inhibitory effects up to the 200 mg/ml dose used.
This work was financially supported by grants from the National Natural Science Foundation of
China (No. 20972166), the National Basic Research Program of China (973 Program) (No.
2009CB526512), the Ministry of Education of China through its 111 & 985 Projects (B08044 & MUC
98503-03-001006), and the Knowledge Innovation Project of the Chinese Academy of Sciences.
Experimental Part
General. Optical rotations: Jasco-DIP-370 automatic digital polarimeter. UV Spectra: Shimadzu-
210A double-beam spectrometer; lmax (log e) in nm. CD Spectra: Jasco-J-715 spectropolarimeter; l (De)
in nm. IR Spectra: Bio-Rad-FTS-135 spectrophotometer; KBr pellets; in cmꢀ1. NMR Spectra: Bruker-
AM-400 and -DRX-500 spectrometers; d in ppm rel. to Me4Si, J in Hz. FAB-MS: VG-Autospec-3000
magnetic-sector instrument; glycerol as matrix in the negative-ion mode and 3-nitrobenzyl alcohol in the
positive-ion mode; in m/z. ESI-MS and HR-ESI-MS: API-Qstar-Pulsar-1 instrument; in m/z. Anal. TLC:
precoated silica-gel-F254 plates (Qingdao Meigao Chemical Co., Qingdao, P. R. China); detection by
spraying with 5% H2SO4 in EtOH, followed by heating. Prep. TLC: silica gel F254 (Qingdao Meigao
Chemical Co.). Column chromatography (CC): silica gel (200 – 300 mesh; Qingdao Meigao Chemical
Co.), C-18 silica gel (40 – 75 mm; Fuji Silysia Chemical Ltd.), D101 resin (Qingdao Meigao Chemical Co.),
Sephadex LH-20 (40 – 70 mm; Amersham Pharmacia Biotech AB, Uppsala, Sweden). HPLC: Agilent-
1200 chromatograph; semi-prep. column Zorbax SB-C18 (9.4 ꢁ 250 mm, 5 mm).
Plant Material. Selaginella moellendorffii was collected from the Jingxi County of the Guangxi
Zhuang Autonomous Region, P. R. China, in June 2008. The plant was identified by Dr. Guang-Wan Hu
(Kunming Institute of Botany, Chinese Academy of Sciences), and a voucher specimen (No. JX0801)
was deposited with the Laboratory of Ethnobotany, Kunming Institute of Botany.
Extraction and Isolation. The air-dried whole plants of S. moellendorffii (6 kg) were exhaustively
extracted with MeOH (6 l; 4, 3, and 3 h, resp.) at 708. The solvent was evaporated, and the remaining
residue (780 g) was partitioned by CC (silica gel) into three fractions, with CHCl3 (A, 32 g), Me2CO (B,
154 g), and MeOH (C, 350 g). Fraction C was separated by CC (D101 resin) into Fr. C1 with H2O (270 g)
and Fr. C2 with MeOH (71 g). Fr. C2 was fractionated by CC (C18, MeOH/H2O 5 :95 ! 95 :5): Fr. C21
with 5% MeOH (18 g), Fr. C22 with 10% MeOH (18 g), Fr. C23 with 20% MeOH (11 g), Fr. C24 with 30%
MeOH (14 g), and Fr. C25 with 50% MeOH (13 g). Fr. C21 was subjected to CC (silica gel, CHCl3/MeOH/
HCOOH 90 :30 :1) and semi-prep. HPLC (MeOH/H2O (containing 0.05% CF3COOH) 10 :90): 11
(29.0 mg; tR 10.3 min), 10 (16.0 mg; tR 12.4 min), and 9 (12.0 mg; tR 18.3 min). Fr. C22 was subjected to CC
(silica gel, CHCl3/MeOH/HCOOH 50 :10 :1, and CHCl3/MeOH 1:1) and semi-prep. HPLC (MeOH/
H2O (containing 0.05% CF3COOH) 20 :80): 12 (17.4 mg; tR 12.8 min) and 14 (11.0 mg; tR 14.0 min).
Fr. C23 was subjected to CC (silica gel, CHCl3/MeOH 1:1; Sephadex LH-20, MeOH: 13 (16.0 mg). Fr. C24
was subjected to CC (silica gel, CHCl3/MeOH/HCOOH 40 :10 :1; Sephadex LH-20, MeOH): 8 (7.0 mg;
Rf 0.6) and 6 (6.2 mg; Rf 0.4), purified by prep. TLC (CHCl3/MeOH/HCOOH, 40 :10 :1), and 5
(13.4 mg; Rf 0.6) and 7 (5.3 mg; Rf 0.7), purified by prep. TLC (AcOEt/MeOH 3: 1). Fr. C25 was
subjected to CC (silica gel, CHCl3/MeOH 50 :1, 20 :1, 10 :1, and 5 :1; Sephadex LH-20, MeOH): 1
(23.0 mg), 4 (28.0 mg), 2 (3.5 mg), and 3 (9.0 mg).

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(7S,8R)-4,9-Dihydroxy-3,3’,5-trimethoxy-4’,7-epoxy-8,5’-neolignan-9’-oic Acid Methyl Ester
(¼(2S,3R)-2,3-Dihydro-2-(4-hydroxy-3,5-dimethoxyphenyl)-3-(hydroxymethyl)-7-methoxybenzofuran-
5-propanoic Acid Methyl Ester; 1): Colorless amorphous solid. [a]2D6 ¼ ꢀ4.4 (c ¼ 0.36, MeOH). UV
(MeOH): 379 (3.07), 290 (3.52), 209 (4.45). CD (c ¼ 0.03, MeOH): 282 (þ0.13), 204 (þ1.22). IR: 3441,
1
1731, 1616, 1519, 1500, 1462, 1213, 1143, 1113, 1033. H- and 13C-NMR: Table 1. FAB-MS (pos.): 419
([M þ H]þ). HR-ESI-MS (pos.): 441.1518 ([M þ Na]þ, C22H26NaO8þ ; calc. 441.1525).
rel-(7R,7’E,8S)-4,9-Dihydroxy-3,3’,5-trimethoxy-4’,7-epoxy-8,5’-neolign-7’-en-9’-oic Acid (¼ rel-
(2E)-3-[(2R,3S)-2,3-Dihydro-2-(4-hydroxy-3,5-dimethoxyphenyl)-3-(hydroxymethyl)-7-methoxybenzo-
furan-5-yl]prop-2-enoic Acid; 2): Colorless amorphous solid. [a]2D5 ¼ ꢀ48.0 (c ¼ 0.083, MeOH). UV
(MeOH): 391 (3.03), 294 (3.63), 207 (4.21). CD (c ¼ 0.05, MeOH): 302 (ꢀ0.52), 238 (þ0.87), 217
(ꢀ1.26), 203 (þ2.26). 1H- and 13C-NMR: Table 1. ESI-MS (neg.): 401 ([M ꢀ H]ꢀ). HR-ESI-MS (neg.):
401.1236 ([M ꢀ H]ꢀ, C21H21Oꢀ8 ; calc. 401.1236).
(7S,8R)-4,9-Dihydroxy-4’,7-epoxy-8’,9’-dinor-8,5’-neolignan-7’-oic Acid (¼(2S,3R)-2,3-Dihydro-3-
(hydroxymethyl)-2-(4-hydroxyphenyl)benzofuran-5-carboxylic Acid; 3): Colorless amorphous solid.
[a]2D6 ¼ þ96.3 (c ¼ 0.52, MeOH). UV (MeOH): 373 (2.96), 258 (3.82), 204 (4.27). CD (c ¼ 0.04, MeOH):
260 (þ 7.54), 229 (ꢀ6.06), 203 (þ6.76). IR: 3397, 1674, 1611, 1600, 1518, 1490, 1250, 1172, 1118, 833, 775.
1H- and 13C-NMR: Table 1. FAB-MS (neg.): 285 ([M ꢀ H]ꢀ). HR-ESI-MS (neg.): 285.0754 ([M ꢀ H]ꢀ,
C16H13Oꢀ5 ; calc. 285.0762).
(7S,8R)-3,3’,5-Trimethoxy-4’,7-epoxy-8,5’-neolignan-4,9,9’-triol (¼(2S,3R)-2,3-Dihydro-2-(4-hy-
droxy-3,5-dimethoxyphenyl)-3-(hydroxymethyl)-7-methoxybenzofuran-5-propanol; 4): Colorless amor-
phous solid. [a]1D6 ¼ꢀ 3.9 (c ¼ 0.80, MeOH). UV (MeOH): 284 (3.44), 208 (4.39). IR: 3425, 1613, 1518,
1499, 1462, 1214, 1142, 1115. 1H- and 13C-NMR: Table 1. ESI-MS (neg.): 425 ([M þ Cl]ꢀ). HR-ESI-MS
(neg.): 425.1370 ([M þ Cl]ꢀ, C21H26ClO7ꢀ ; calc. 425.1367).
rel-(7R,8S)-3,3’,5-Trimethoxy-4’,7-epoxy-8,5’-neolignan-4,9,9’-triol 4-b-d-Glucopyranoside (¼4-
[(2S,3R)-2,3-Dihydro-3-(hydroxymethyl)-5-(3-hydroxypropyl)-7-methoxybenzofuran-2-yl]-2,6-dimeth-
oxyphenyl b-d-Glucopyranoside; 5): Colorless amorphous solid. [a]1D6 ¼ ꢀ33.0 (c ¼ 0.67, MeOH). UV
(MeOH): 281 (3.39), 208 (4.60). CD (c ¼ 0.03, MeCN): 206 (ꢀ1.89). IR: 3424, 1599, 1501, 1463, 1422,
1
1124, 1069. H- and 13C-NMR: Table 2. ESI-MS (neg.): 587 ([M þ Cl]ꢀ). HR-ESI-MS (neg.): 587.1883
([M þ Cl]ꢀ, C27H36ClO1ꢀ2 ; calc. 587.1895).
rel-(7R,8S)-3,3’,5-Trimethoxy-4’,7-epoxy-8,5’-neolignan-4,9,9’-triol 9-b-d-Glucopyranoside (¼ rel-
[(2R,3S)-2,3-Dihydro-2-(4-hydroxy-3,5-dimethoxyphenyl)-5-(3-hydroxypropyl)-7-methoxybenzofuran-
3-yl]methyl b-d-Glucopyranoside; 6): Colorless amorphous solid. [a]2D6 ¼ ꢀ19.3 (c ¼ 0.20, MeOH). UV
(MeOH): 377 (3.00), 291 (3.52), 208 (4.26). CD (c ¼ 0.075, MeOH): 310 (ꢀ0.28), 272 (ꢀ0.10), 246
1
(þ0.30), 229 (ꢀ0.18), 206 (ꢀ0.75). IR: 3423, 2930, 1603, 1516, 1501, 1463, 1116, 1075, 1043. H- and
13C-NMR: Table 2. ESI-MS (neg.): 587 ([M þ Cl]ꢀ). HR-ESI-MS (neg.): 587.1881 ([M þ Cl]ꢀ,
C27H36ClOꢀ12 ; calc. 587.1895).
3,3’,5-Trimethoxy-4’,7-epoxy-8,5’-neolign-7-ene-4,9,9’-triol 9-b-d-Glucopyranoside (¼ [2-(4-Hy-
droxy-3,5-dimethoxyphenyl)-5-(3-hydroxypropyl)-7-methoxybenzofuran-3-yl]methyl b-d-Glucopyrano-
side; 7): Colorless amorphous solid. [a]2D8 ¼ ꢀ46.6 (c ¼ 0.27, MeOH). UV (MeOH): 307 (4.04), 219
(4.21), 204 (4.21). IR: 3421, 1614, 1516.6, 1462, 1219, 1110. 1H- and 13C-NMR: Table 2. FAB-MS (neg.):
549 ([M ꢀ H]ꢀ). HR-ESI-MS (neg.): 549.1956 ([M ꢀ H]ꢀ, C27H33O1ꢀ2 ; calc. 549.1972).
3,3’,5,5’-Tetramethoxy-8,4’-oxyneolignan-4,9,9-triol 4-b-d-Glucopyranoside (¼4-{3-Hydroxy-2-[4-(3-
hydroxypropyl)-2,6-dimethoxyphenoxy]propyl}-2,6-dimethoxyphenyl b-d-Glucopyranoside; 8): Color-
less amorphous solid. [a]2D6 ¼ ꢀ25.0 (c ¼ 0.35, MeOH). UV (MeOH): 269 (3.56), 207 (4.57). IR: 3419,
1
1594, 1504, 1462, 1422, 1243, 1226, 1125, 1067. H- and 13C-NMR: Table 2. FAB-MS (neg.): 583 ([M ꢀ
H]ꢀ). HR-ESI-MS (neg.): 619.2145 ([M þ Cl]ꢀ, C28H40ClO1ꢀ3 ; calc. 619.2157).
myo-Inositol 1-Caffeate (¼ myo-Inositol 1-[(2E)-3-(3,4-Dihydroxyphenyl)prop-2-enoate]; 9): Color-
less amorphous solid. [a]2D6 ¼ ꢀ6.4 (c ¼ 0.14, MeOH). UV (MeOH): 329 (3.96), 244 (3.78), 234 (3.78),
1
218 (3.94). IR: 3407, 1688, 1631, 1604, 1523, 1445, 1281, 1182, 1116, 1034. H- and 13C-NMR: Table 3.
FAB-MS (neg.): 341 ([M ꢀ H]ꢀ). HR-ESI-MS (neg.): 341.0878 ([M ꢀ H]ꢀ, C15H17Oꢀ9 ; calc. 341.0872).
myo-Inositol 6-Caffeate (¼ myo-Inositol 6-[(2E)-3-(3,4-Dihydroxyphenyl)prop-2-enoate]; 10): Col-
orless amorphous solid. [a]2D7 ¼ ꢀ7.9 (c ¼ 0.24, MeOH). UV (MeOH): 327 (3.68), 297 (3.61), 206 (3.83).

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Helvetica Chimica Acta – Vol. 93 (2010)
IR: 3345, 1690, 1633, 1607, 1519, 1460, 1401, 1380, 1260, 1180, 1112, 1040. 1H- and 13C-NMR: Table 3. ESI-
MS (neg.): 341 ([M ꢀ H]ꢀ). HR-ESI-MS (neg.): 341.0874 ([M ꢀ H]ꢀ, C15H17Oꢀ9 ; calc. 341.0872).
myo-Inositol 5-Caffeate (¼ myo-Inositol 5-[(2E)-3-(3,4-Dihydroxyphenyl)prop-2-enoate]; 11): Col-
orless amorphous solid. UV (MeOH): 327 (3.75), 300 (3.66), 216 (3.79). IR: 3372, 1688, 1632, 1598,
1
1528, 1445, 1116, 1042, 718. H- and 13C-NMR: Table 3. ESI-MS (neg.): 341 ([M ꢀ H]ꢀ). HR-ESI-MS
(neg.): 341.0879 ([M ꢀ H]ꢀ, C15H17O9ꢀ ; calc. 341.0872).
Paucine 3’-b-d-Glucopyranoside (¼ N-(4-Aminobutyl)caffeamide 3’-b-d-Glucopyranoside ¼ (2E)-N-
(4-Aminobutyl)-3-[3-(b-d-glucopyranosyloxy)-4-hydroxyphenyl]prop-2-enamide; 12): Colorless amor-
phous solid. [a]2D6 ¼ ꢀ23.3 (c ¼ 0.35, MeOH). UV (MeOH): 387 (2.18), 373 (2.30), 312 (3.64), 293 (3.61),
230 (3.60), 216 (3.62). IR: 3439, 1639, 1630, 1547, 1511, 1250, 1173, 1033. 1H-NMR (CD3OD, 500 MHz):
3.33 (t, J ¼ 7.1, CH2CH2CH2CH2NCO), 1.66 – 1.73 (m, CH2CH2CH2CH2NCO), 1.60 – 1.66 (m,
CH2CH2CH2CH2NCO), 2.96 (t, J ¼ 6.6, CH2CH2CH2CH2NCO), 7.46 (s, HꢀC(2’)); 6.84 (d, J ¼ 8.0,
HꢀC(5’)); 7.12 (d, J ¼ 8.0, HꢀC(6’)); 7.43 (d, J ¼ 16.0, HꢀC(3)); 6.41 (d, J ¼ 16.0, HꢀC(2)); 4.80 (d, J ¼
7.7, H ꢀC(1’’)); 3.47 – 3.54 (m, Hꢀ(2’’)); 3.46 – 3.52 (m, HꢀC(3’’)); 3.38 (t, J ¼ 9.0, HꢀC(4’’)); 3.43 – 3.50
(m, HꢀC(5’’)); 3.94 (d, J ¼ 11.5, HaꢀC(6’’)); 3.71 (dd, J ¼ 11.5, 6.0, HbꢀC(6’’)). 13C-NMR (CD3OD,
125 MHz): 39.6 (t, C(1) (Bu)); 25.9 (t, C(2) (Bu)); 27.6 (t, C(3) (Bu)); 40.4 (t, C(4) (Bu)); 128.4 (s, C(1’));
117.4 (d, C(2’)); 147.1 (s, C(3’)); 150.4 (s, C(4’)); 117.4 (d, C(5’)); 125.7 (d, C(6’)); 141.9 (d, C(3)); 119.0 (d,
C(2)); 169.3 (s, C(1)); 104.2 (d, C(1’’)); 74.9 (d, C(2’’)); 77.6 (d, C(3’’)); 71.5 (d, C(4’’)); 78.5 (d, C(5’’));
62.6 (t, C(6’’)). ESI-MS (pos.): 413 ([M þ H]þ). HR-ESI-MS (pos.): 413.1933 ([M þ H]þ, C19H29N2Oþ8 ;
calc. 413.1923).
Acid Hydrolysis of 5. Compound 5 (5 mg) was dissolved in 2n HCl (2 ml) and hydrolyzed (4 h) at
908. The acidic soln. was concentrated, and the residue separated by CC (silica gel, CHCl3/MeOH/H2O
40 :10 :0.5): 0.8 mg of d-glucose, detected by TLC and optical rotation. [a]2D2 ¼ þ28.2 (c ¼ 0.08, H2O).
Biological Testing. The activity of compounds 7 and 11, and N1-cis-p-coumaroylagmatine (14) against
K562 cells were measured by the MTT method with adriamycin as pos. control (IC50 ¼ 0.32 mm) [24], and
that of compound 12 and paucine (13) against AChE was assessed by Ellmanꢂs method with tacrine as
pos. control (IC50 ¼ 0.20 mm) [25], resp.
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Received March 22, 2010