Cytotoxic activity of dietary lignan and its derivatives: Structure-cytotoxic activity relationship of dihydroguaiaretic acid

Article abstract of DOI:10.1021/jf5010572

Cytotoxic activities of synthesized lignan derivatives were estimated by WST-8 reduction assay against HL-60 and HeLa cells to show the structure-activity relationship. The activities of some effective compounds were examined against Colon 26 and Vero cel

Full text of DOI:10.1021/jf5010572

Article
pubs.acs.org/JAFC
Cytotoxic Activity of Dietary Lignan and Its Derivatives: Structure−
Cytotoxic Activity Relationship of Dihydroguaiaretic Acid
Tuti Wukirsari,† Hisashi Nishiwaki,† Kosuke Nishi,† Takuya Sugahara,†,‡ Koichi Akiyama,§
Taro Kishida,†,‡ and Satoshi Yamauchi*,†,‡
§
†Faculty of Agriculture and Integrated Center for Sciences, Tarumi Station, Ehime University, 3-5-7 Tarumi, Matsuyama, Ehime
790-8566, Japan
‡South Ehime Fisheries Research Center, 1289-1 Funakoshi, Ainan, Ehime 798-4292, Japan
S
* Supporting Information
ABSTRACT: Cytotoxic activities of synthesized lignan derivatives were estimated by WST-8 reduction assay against HL-60 and
HeLa cells to show the structure−activity relationship. The activities of some effective compounds were examined against Colon
26 and Vero cells. Dietary secoisolariciresinol (SECO, 1) and its metabolite, 9,9′-anhydrosecoisolariciresinol (2), did not show
the cytotoxic activity. On the other hand, all stereoisomers of dihydroguaiaretic acid (DGA, 9,9′-dehydroxysecoisolariciresinol,
3−5) exhibited the activity (IC50: around 30 μM). The IC50 value of (8R,8′R)-9-butyl DGA derivative 13 was around 6 μM. This
fact means that the hydrophobic group was advantageous for higher activity at 9- and 9′-positions. By the evaluation of the effect
of 7and 7′-aryl group on the activity, we discovered the highest activity of (8R,8′R)-7-(3-hydroxy-4-methoxyphenyl)-7′-(2-
ethoxyphenyl) DGA derivative 47 showing around 1 μM of IC50 value, which is about 24-fold higher activity than that of natural
(8R,8′R)-DGA. The derivative of dietary lignan showed the high cytotoxic activity.
KEYWORDS: lignan, cytotoxic activity, secoisolariciresinol, dihydroguaiaretic acid
To achieve this project, the synthetic route to 9- and 9′-
derivatives 6−15 was also developed (Figure 1).
INTRODUCTION
■
Lignans are widely distributed in grains, vegetables, and fruits.1
Applications of dietary and medicinal lignan to the health and
food industry, including inhibitory activity of cAMP by
dibenzylbutyrolactone and furofuran lignans,2 inhibitor and
antagonist of PAF by neoliganan kadsurenone and (+)-dihy-
droguaiaretic acid,2 biological activity as polyphenol by
furofuran lignan,3 IgE-suppressive activity−structure relation-
ship of (−)-matairesinol,4 immunomodulatroy effect of
(−)-matairesinol,5 effect of secoisolariciresinol (1) on 3T3-L1
adipocytes,6 inhibition of the discolorlation of yellowtail dark
muscle by (−)-matairesinol,7 and cytotoxic activity−structure
relationship of morinol A8 have been reported. Dietary lignans
have served as lead structures for new clinical candidates from
natural products.
The effect of the structure between the 8- and 8′-position of
butane-type lignan on the cytotoxic activity11 and butane-type
lignan as an apoptosis inducer12 have been reported. However,
structure−activity relationships on 9- and 9′-positions and 7-
and 7′-aromatic rings have been unknown. Our purpose is to
find out the most advantageous substituent for cytotoxic
activity on each portion. The results could give information
about the structure−activity relationship of dietary and
metabolite lignans, which contribute to development of new
clinical medicine based on dietary compounds.
MATERIALS AND METHODS
■
Chemicals. (−)-Secoisolariciresinol (SECO) (1),13 9,9′-
anhydrosecoisolariciresinol (2),14 and stereoisomers of dihy-
droguaiaretic acid (3−5)15,16 were synthesized by the
previously described method. The derivatives 6−15 were
synthesized by the new synthetic method shown in Scheme 1,
and their procedures are described in Supporting Information.
The derivatives 16−59 were synthesized by the previously
described method15 with modification. NMR data of 16−18,
20−24, 26−30, and 32−43 agreed with those of the
literature.17,18 The [α]D values of 20−22, 26−28, and 32−34
were opposite but equal amounts to the corresponding
enantiomers in the literature.18 Optical rotation values were
On the other hand, the research on metabolism of plant
lignans to many types of lignans and lignan derivatives are also
continuing,9 suggesting that new lignan metabolites would be
discovered in the future. This fact suggests that the effect of
each lignan and its derivative on health should be clarified by
structure−activity relationship research because safety of food is
focused.
In this study, cytotoxic activity of dietary secoisolariciresinol
(1) and its reductive products, dihydroguaiaretic acid stereo-
isomers (3-5), were examined as a first stage. The effect of the
stereochemistry was compared and discussed for the first time.
The activity of 9,9′-anhydrosecoisolariciresinol (2), which is
proposed as a metabolite compound from secoisolariciresinol,10
was also estimated. As a next stage, we designed the 9- and 9′-
drivatives 6−15 and the aromatic derivatives 16−59 to clarify
the structure−cytotoxic activity relationship for the first time.
Received: March 2, 2014
Revised: May 13, 2014
Accepted: May 19, 2014
Published: May 19, 2014
© 2014 American Chemical Society
5305
dx.doi.org/10.1021/jf5010572 | J. Agric. Food Chem. 2014, 62, 5305−5315

Journal of Agricultural and Food Chemistry
Article
Figure 1. Butane-type lignans and their derivatives.
measured with a Horiba SEPA-200 instrument. NMR data were
obtained with a JNM-ECS400 spectrometer, using tetrame-
thylsilane and trifluoroacetic acid as a standard (0 ppm) for 1H
NMR and (−76.5 ppm) for 19F-NMR, respectively. EIMS data
were measured with a JMS-MS700 V spectrometer. The
synthetic methods are described in Supporting Information.
The numbering of compounds follows the nomenclature of
lignans.19
J = 9.5, 5.4 Hz), 3.30 (6H, s), 3.35 (2H, dd, J = 9.5, 5.4 Hz),
3.76 (6H, s), 5.58 (2H, s), 6.51 (2H, d, J = 1.9 Hz), 6.59 (2H,
dd, J = 7.9, 1.9 Hz), 6.78 (2H, d, J = 7.9 Hz); 13C NMR (100
MHz, CDCl3) δ 35.1, 40.6, 55.7, 58.7, 72.7, 111.3, 113.9, 121.9,
132.9, 143.6, 146.4; MS (EI) m/z 390 (M+, 41), 358 (21), 189
(32), 137 (100); HRMS (EI) m/z calcd for C22H30O6
390.2043, found 390.2050.
(8R,8′R)-4,4′-Dihydroxy-3,3′-dimethoxy-9a,9′a-dihomo-
25
lignane-9a,9′a- dinitrile (8). Colorless oil; [α]D +40 (c 0.5,
(8R,8′R)-4,4′-Dihydroxy-3,3′-dimethoxylignane-9,9′-diyl
1
diacetate (6). Colorless crystals; mp 108−110 °C; [α]D25 −30
CHCl3); H NMR (400 MHz, CDCl3) δ 2.24 (2H, m), 2.36−
1
2.38 (4H, m), 2.57 (2H, dd, J = 14.0, 9.4 Hz), 2.98 (2H, dd, J =
14.0, 5.0 Hz), 3.88 (6H, s), 5.58 (2H, s), 6.64 (2H, d, J = 1.9
Hz), 6.66 (2H, dd, J = 7.9, 1.9 Hz), 6.87 (2H, d, J = 7.9 Hz);
13C NMR (100 MHz, CDCl3) δ 19.0, 36.3, 39.3, 56.0, 111.0,
114.7, 118.1, 121.7, 129.6, 144.7, 146.9; EIMS m/z 380 (M+,
43), 137 (100), 122 (12); HRMS (EI) m/z calcd for
C22H24O4N2 380.1737, found 380.1739.
(c 1.3, CHCl3); H NMR (400 MHz, CDCl3) δ 2.07 (6H, s),
2.07−2.11 (2H, overlapped), 2.61 (4H, m), 3.78 (6H, s), 4.03
(2H, dd, J = 11.4, 5.7 Hz), 4.20 (2H, dd, J = 11.4, 5.9 Hz), 5.62
(2H, br. s), 6.47 (2H, d, J = 2.0 Hz), 6.55 (2H, dd, J = 8.0, 2.0
Hz), 6.79 (2H, d, J = 8.0 Hz); 13C NMR (100 MHz, CDCl3) δ
21.0, 35.1, 39.6, 55.8, 64.5, 111.2, 114.2, 121.7, 131.5, 144.0,
146.5, 171.1; MS (EI) m/z 446 (M+, 42), 189 (19), 137 (100);
HRMS (EI) m/z calcd for C24H30O8 446.1941, found
446.1940.
(8R,8′R)-3,3′-Dimethoxy-4,4′,9-liganetriol (9). Colorless
oil; [α]D −24 (c 1.0, CHCl3); 1H NMR (400 MHz,
25
(8R,8′R)- 3,3′,9,9′-Tetramethoxy-4,4′-lignanediol (7). Col-
CDCl3) δ 0.90 (3H, d, J = 6.9 Hz), 1.40−1.70 (1H, br), 1.77
(1H, m), 1.88 (1H, dq, J = 6.9, 3.3 Hz), 2.39 (1H, dd, J = 13.8,
8.1 Hz), 2.53 (1H, dd, J = 13.8, 7.6 Hz), 2.65 (1H, dd, J = 13.8,
25
1
orless oil; [α]D −27 (c 1.1, CHCl3); H NMR (400 MHz,
CDCl3) δ 2.01 (2H, m), 2.61 (4H, d, J = 7.3 Hz), 3.28 (2H, dd,
5306
dx.doi.org/10.1021/jf5010572 | J. Agric. Food Chem. 2014, 62, 5305−5315

Journal of Agricultural and Food Chemistry
Article
Scheme 1. Syntheses of 9- and 9′-Derivatives of (8R,8′R)-Dihydroguaiaretic Acid
7.0 Hz), 2.67 (1H, dd, J = 13.8, 7.5 Hz), 3.57 (1H, dd, J = 10.9,
6.6 Hz), 3.72 (1H, dd, J = 10.9, 5.3 Hz), 3.77 (3H, s), 3.78 (3H,
s), 5.64 (2H, br s), 6.50 (1H, d, J = 1.9 Hz), 6.54 (1H, d, J = 1.9
Hz), 6.57 (1H, dd, J = 8.0, 1.8 Hz), 6.61 (1H, dd, J = 8.0, 1.8
Hz), 6.78 (1H, d, J = 8.0 Hz), 6.80 (1H, d, J = 8.0 Hz); 13C
NMR (100 MHz, CDCl3) δ 15.6, 35.0, 35.3, 40.5, 45.8, 55.75,
55.78, 63.0, 111.29, 111.34, 114.0, 114.1, 121.67, 121.71, 132.9,
133.2, 143.6, 143.7, 146.4, 146.5; MS (EI) m/z 346 (M+, 11),
137 (100), 122 (14); HRMS (EI) m/z calcd for C20H26O5
346.17808, found 346.17811.
(8R,8′R)-3,3′-Dimethoxy-4,4′-dihydroxyligan-9-yl acetate
25 1
(10). Colorless oil; [α]D −28 (c 0.8, CHCl3); H NMR
(400 MHz, CDCl3) δ 0.91 (3H, d, J = 7.1 Hz), 1.83 (1H, dq, J
= 7.1, 3.1 Hz), 1.94 (1H, m), 2.06 (3H, s), 2.40 (1H, dd, J =
13.7, 7.8 Hz), 2.52 (1H, dd, J = 13.9, 8.2 Hz), 2.61 (1H, dd, J =
13.7, 6.9 Hz), 2.62 (1H, dd, J = 13.9, 6.5 Hz), 3.78 (3H, s), 3.79
(3H, s), 4.00 (1H, dd, J = 11.2, 6.7 Hz), 4.19 (1H, dd, J = 11.2,
5.8 Hz), 5.53 (1H, s), 5.55 (1H, s), 6.47−6.48 (2H, m), 6.55
(1H, dd, J = 8.0, 2.0 Hz), 6.57 (1H, dd, J = 7.9, 1.9 Hz), 6.79
(1H, d, J = 8.0 Hz), 6.80 (1H, d, J = 7.9 Hz); 13C NMR (100
MHz, CDCl3) δ 15.2, 21.1, 35.0, 35.4, 40.6, 41.9, 55.7, 55.8,
5307
dx.doi.org/10.1021/jf5010572 | J. Agric. Food Chem. 2014, 62, 5305−5315

Journal of Agricultural and Food Chemistry
Article
64.7, 111.2, 111.3, 114.0, 114.1, 121.7, 132.2, 132.9, 143.7,
143.8, 146.3, 146.4, 171.3; MS (EI) m/z 388 (M+, 25), 137
(100); HRMS (EI) m/z calcd for C22H28O6 388.1886, found
388.1885.
HRMS (EI) m/z calcd for C23H32O4 372.2302, found
372.2305.
(8R,8′R)-9-(N,N-Dimethylamino)-3,3′-dimethoxylignane-
25
1
4,4′-diol (15). Colorless oil; [α]D −59 (c 0.4, CHCl3); H
NMR (400 MHz, CDCl3) δ 0.85 (3H, d, J = 7.0 Hz), 1.71 (1H,
m), 1.86 (1H, dq, J = 7.0, 2.3 Hz), 2.15−2.24 (2H, overlapped),
2.20 (6H, s), 2.33 (1H, dd, J = 13.8, 9.7 Hz), 2.39 (1H, dd, J =
13.7, 7.1 Hz), 2.49 (1H, dd, J = 13.7, 8.4 Hz), 2.77 (1H, dd, J =
13.8, 5.1 Hz), 3.71 (3H, s), 3.76 (3H, s), 6.39 (1H, d, J = 1.8
Hz), 6.46 (1H, d, J = 1.8 Hz), 6.51 (1H, dd, J = 8.0, 1.8 Hz),
6.55 (1H, dd, J = 8.0, 1.8 Hz), 6.74 (1H, d, J = 8.0 Hz), 6.77
(1H, d, J = 8.0 Hz); 13C NMR (100 MHz, CDCl3) δ 14.8, 33.9,
36.3, 40.0, 40.2, 46.2, 55.6, 55.7, 60.1, 111.1, 111.3, 113.7,
113.8, 121.7, 121.8, 133.3, 133.5, 143.49, 143.54, 146.3; MS
(EI) m/z 373 (M+, 100), 164 (20), 137 (43); HRMS (EI) m/z
calcd for C22H31O4N 373.2254, found 373.2260.
(8R,8′R)-3,3′,9-Trimethoxy-4,4′-liganediol (11). Colorless
oil; [α]D25 −30 (c 1.0, CHCl3); 1H NMR (400 MHz, CDCl3) δ
0.87 (3H, d, J = 6.9 Hz), 1.80−1.88 (2H, m), 2.36 (1H, dd, J =
13.8, 7.8 Hz), 2.49 (1H, dd, J = 13.8, 7.7 Hz), 2.65 (1H, dd, J =
13.8, 7.1 Hz), 2.66 (1H, dd, J = 13.8, 7.0 Hz), 3.25 (1H, dd, J =
9.4, 6.2 Hz), 3.32 (3H, s), 3.40 (1H, dd, J = 9.4, 5.3 Hz), 3.765
(3H, s), 3.772 (3H, s), 5.53 (1H, s), 5.55 (1H, s), 6.48 (1H, d, J
= 1.9 Hz), 6.52 (1H, d, J = 1.9 Hz), 6.57 (1H, dd, J = 8.1, 1.9
Hz), 6.59 (1H, dd, J = 8.1, 1.9 Hz), 6.77 (1H, d, J = 8.1 Hz),
6.79 (1H, d, J = 8.1 Hz); 13C NMR (100 MHz, CDCl3) δ 15.4,
34.8, 35.3, 40.7, 43.3, 55.70, 55.74, 58.8, 72.9, 111.28, 111.32,
113.9, 121.79, 121.80, 133.0, 133.4, 143.5, 143.6, 146.3, 146.4;
MS (EI) m/z 360 (M+, 14), 137 (100), 122 (14); HRMS (EI)
m/z calcd for C21H28O5 360.1937, found 360.1929.
(8R,8′R)-2-Butoxy-3′-methoxy-4′-lignanol (19). Colorless
oil; [α]D25 −42 (c 0.8, CHCl3); 1H NMR (400 MHz, CDCl3) δ
0.81 (3H, d, J = 6.9 Hz), 0.85 (3H, d, J = 6.7 Hz), 0.96 (3H, t, J
= 7.4 Hz), 1.46 (2H, m), 1.68 (2H, m), 1.75 (1H, m), 1.85
(1H, m), 2.35 (1H, dd, J = 13.6, 8.4 Hz), 2.38 (1H, dd, J = 12.9,
8.2 Hz), 2.57 (1H, dd, J = 13.6, 6.8 Hz), 2.69 (1H, dd, J = 12.9,
6.2 Hz), 3.78−3.84 (1H, overlapped), 3.80 (3H, s), 3.90 (1H,
m), 5.43 (1H, s), 6.53 (1H, d, J = 1.8 Hz), 6.58 (1H, dd, J = 7.9,
1.8 Hz), 6.78 (2H, br d, J = 7.9 Hz), 6.82 (1H, dd, J = 7.7, 7.3,
1.4 Hz), 7.03 (1H, dd, J = 7.3, 1.5 Hz), 7.12 (1H, ddd, J = 7.8,
7.7, 1.8 Hz); 13C NMR (100 MHz, CDCl3) δ 13.9, 14.1, 14.2,
19.4, 31.5, 35.9, 36.1, 38.6, 41.1, 55.8, 67.3, 110.9, 111.3, 113.9,
119.8, 121.6, 126.8, 130.2, 130.8, 133.9, 143.4, 146.2, 157.2; MS
(EI) m/z 356 (M+, 88), 163 (34), 137 (81), 107 (29), 59
(100); HRMS (EI) m/z calcd for C23H32O3 356.2351, found
356.2340.
(8R,8′R)-9-Fluoro-3,3′-dimethoxy-4,4′-lignanediol (12).
25
1
Colorless oil; [α]D +20 (c 0.2, CHCl3); H NMR (400
MHz, CDCl3) δ 0.94 (3H, d, J = 6.8 Hz), 1.85−1.98 (2H, m),
2.43 (1H, dd, J = 13.8, 7.6 Hz), 2.56 (1H, dd, J = 13.6, 7.4 Hz),
2.62−2.69 (2H, m), 3.798 (3H, s), 3.803 (3H, s), 4.45 (2H, m),
5.46 (1H, s), 5.48 (1H, s), 6.49 (1H, br s), 6.51 (1H, br s), 6.58
(1H, br d, J = 8.1 Hz), 6.60 (1H, br d, J = 7.7 Hz), 6.79 (1H,
dd, J = 7.7, 1.2 Hz), 6.81 (1H, dd, J = 8.1, 1.2 Hz); 13C NMR
(100 MHz, CDCl3) δ 15.5, 34.3, 34.8 (d, J = 77.8 Hz), 40.9,
43.8 (d, J = 17.2 Hz), 55.7, 55.8, 84.0 (d, J = 167.7 Hz), 111.15,
111.21, 113.9, 114.0, 121.7, 132.0, 132.8, 143.6, 143.8, 146.3,
146.4; 19F NMR (376 MHz, CDCl3) δ −226.4; MS (EI) m/z
(%) 348 (M+, 57), 137 (100); HRMS (EI) m/z calcd for
C20H25O4F 348.1737, found 348.1728.
(8R,8′R)-3-Butoxy-3′-methoxy-4′-lignanol (25). Colorless
oil; [α]D25 −28 (c 0.6, CHCl3); 1H NMR (400 MHz, CDCl3) δ
0.83 (6H, d, J = 6.7 Hz), 0.98 (3H, t, J = 7.4 Hz), 1.49 (2H, m),
1.72−1.82 (4H, m), 2.37 (1H, dd, J = 13.6, 8.1 Hz), 2.41 (1H,
dd, J = 13.5, 8.1 Hz), 2.55 (1H, dd, J = 13.6, 6.7 Hz), 2.59 (1H,
dd, J = 13.5, 6.7 Hz), 3.82 (3H, s), 3.91 (2H, t, J = 6.5 Hz), 5.45
(1H, s), 6.54 (1H, d, J = 1.7 Hz), 6.59 (1H, dd, J = 8.1, 1.7 Hz),
6.63 (1H, m), 6.67 (1H, br d, J = 7.5 Hz), 6.70 (1H, m), 6.80
(1H, d, J = 8.1 Hz), 7.14 (1H, dd, J = 7.8, 7.8 Hz); 13C NMR
(100 MHz, CDCl3) δ 13.9, 14.0, 19.3, 31.5, 37.6, 37.9, 41.1,
41.5, 55.8, 67.5, 111.3, 111.5, 113.9, 115.3, 121.3, 121.6, 128.9,
133.5, 143.3, 143.5, 146.2, 159.0; MS (EI) m/z 356 (M+, 23),
164 (62), 137 (100), 107 (74), 58 (62); HRMS (EI) m/z calcd
for C23H32O3 356.2351, found 356.2342.
(8R,8′R)-3,3′-Dimethoxy-9a,9b,9c,9d-tetrahomolignane-
25
1
4,4′-diol (13). Colorless oil; [α]D −37 (c 0.9, CHCl3); H
NMR (400 MHz, CDCl3) δ 0.83 (3H, d, J = 6.9 Hz), 0.89 (3H,
t, J = 7.2 Hz), 1.10 (1H, m), 1.24−1.44 (7H, m), 1.49 (1H, m),
1.73 (1H, dq, J = 6.9, 2.3 Hz), 2.33 (1H, dd, J = 13.7, 2.4 Hz),
2.35 (1H, dd, J = 13.7, 4.1 Hz), 2.47 (1H, dd, J = 13.7, 7.8 Hz),
2.57 (1H, dd, J = 13.7, 5.9 Hz), 3.75 (3H, s), 3.77 (3H, s), 5.49
(2H, s), 6.41 (1H, d, J = 1.8 Hz), 6.44 (1H, d, J = 1.8 Hz), 6.51
(1H, dd, J = 8.0, 1.9 Hz), 6.55 (1H, dd, J = 8.0, 1.9 Hz), 6.77
(1H, d, J = 8.0 Hz), 6.78 (1H, d, J = 8.0 Hz); 13C NMR (100
MHz, CDCl3) δ 14.1, 14.8, 22.7, 27.7, 28.9, 32.4, 35.4, 37.5,
40.3, 42.3, 55.65, 55.73, 111.19, 111.24, 113.79, 113.83, 121.69,
121.73, 133.7, 133.8, 143.4, 146.2, 146.3; MS (EI) m/z 386
(M+, 49), 137 (100); HRMS (EI) m/z calcd for C24H34O4
386.2458, found 386.2449.
(8R,8′R)-4-Butoxy-3′-methoxy-4′-lignanol (31). Colorless
oil; [α]D25 −18 (c 0.7, CHCl3); 1H NMR (400 MHz, CDCl3) δ
0.81 (3H, d, J = 6.8 Hz), 0.82 (3H, d, J = 6.8 Hz), 0.98 (3H, t, J
= 7.3 Hz), 1.49 (2H, m), 1.70−1.79 (4H, m), 2.36 (1H, dd, J =
13.6, 3.2 Hz), 2.38 (1H, dd, J = 13.5, 3.2 Hz), 2.55 (1H, dd, J =
13.5, 3.5 Hz), 2.56 (1H, dd, J = 13.6, 3.5 Hz), 3.82 (3H, s), 3.93
(2H, t, J = 6.5 Hz), 5.46 (1H, s), 6.54 (1H, d, J = 1.8 Hz), 6.57
(1H, dd, J = 8.1, 1.8 Hz), 6.78 (2H, d, J = 8.5 Hz), 6.80 (1H, d,
J = 8.1 Hz), 6.98 (2H, d, J = 8.5 Hz); 13C NMR (100 MHz,
CDCl3) δ 13.9, 19.3, 31.5, 37.86, 37.91, 40.5, 41.1, 55.8, 67.7,
111.3, 113.9, 114.1, 121.6, 129.8, 133.5, 133.6, 143.5, 146.2,
157.2; MS (EI) m/z 356 (M+, 12), 208 (39), 137 (99), 107
(74), 58 (100); HRMS (EI) m/z calcd for C23H32O3 356.2351,
found 356.2341.
(8R,8′R)-3,3′-Dimethoxy-9a,9b,9b-trihomoligane-4,4′-diol
25
(14). Colorless crystals; mp 70−71 °C; [α]D −41 (c 0.4,
1
CHCl3); H NMR (400 MHz, CDCl3) δ 0.83 (3H, d, J = 6.9
Hz), 0.84 (3H, d, J = 6.5 Hz), 0.89 (3H, d, J = 6.6 Hz), 1.08
(1H, ddd, J = 18.2, 4.9, 4.9 Hz), 1.18 (1H, ddd, J = 18.2, 3.9, 3.9
Hz), 1.58−1.66 (2H, m), 1.75 (1H, dq, J = 6.6, 2.2 Hz), 2.31
(1H, dd, J = 14.0, 9.6 Hz), 2.34 (1H, dd, J = 14.0, 6.8 Hz), 2.43
(1H, dd, J = 13.8, 8.0 Hz), 2.56 (1H, dd, J = 13.8, 5.4 Hz), 3.74
(3H, s), 3.77 (3H, s), 5.44 (1H, s), 5.45 (1H, s), 6.40 (1H, d, J
= 1.9 Hz), 6.43 (1H, d, J = 1.9 Hz), 6.51 (1H, dd, J = 8.0, 1.9
Hz), 6.55 (1H, dd, J = 8.0, 1.9 Hz), 6.76 (1H, d, J = 8.0 Hz),
6.78 (1H, d, J = 8.0 Hz); 13C NMR (100 MHz, CDCl3) δ 14.6,
22.3, 24.0, 25.5, 35.1, 37.6, 38.4, 39.6, 40.2, 55.6, 55.7, 111.16,
111.21, 113.7, 113.8, 121.7, 121.8, 133.6, 133.8, 143.4, 146.2,
146.3; MS (EI) m/z 372 (M+, 33), 137 (100), 122 (13);
25
(8R,8′R)-4-Methoxy-3-lignanol (44). Colorless oil; [α]D
−24 (c 0.5, CHCl3); 1H NMR (400 MHz, CDCl3) δ 0.81 (3H,
d, J = 6.7 Hz), 0.82 (3H, d, J = 6.7 Hz), 1.75−1.83 (2H, m),
5308
dx.doi.org/10.1021/jf5010572 | J. Agric. Food Chem. 2014, 62, 5305−5315

Journal of Agricultural and Food Chemistry
Article
2.33 (1H, dd, J = 13.5, 5.6 Hz), 2.41 (1H, dd, J = 13.5, 8.7 Hz),
2.57 (1H, dd, J = 13.5, 6.1 Hz), 2.65 (1H, dd, J = 13.5, 6.0 Hz),
3.86 (3H, s), 5.54 (1H, s), 6.56 (1H, dd, J = 8.1, 1.9 Hz), 6.69
(1H, d, J = 1.9 Hz), 6.73 (1H, d, J = 8.1 Hz), 7.10 (2H, d, J =
7.3 Hz), 7.16 (1H, dd, J = 7.3, 7.3 Hz), 7.23−7.27 (2H, m); 13C
NMR (100 MHz, CDCl3) δ 13.9, 14.0, 38.2, 38.3, 40.7, 41.4,
56.0, 110.4, 115.1, 120.3, 125.6, 128.1, 129.0, 135.0, 141.7,
144.5, 145.2; MS (EI) m/z 284 (M+, 69), 137 (100), 101 (75),
91 (30); HRMS (EI) m/z calcd for C19H24O2 284.1776, found
284.1768.
31.5, 35.8, 36.6, 38.9, 40.7, 56.0, 67.3, 110.3, 110.9, 115.1,
119.8, 120.3, 126.7, 130.2, 130.8, 135.4, 144.5, 145.2, 157.1; MS
(EI) m/z 356 (M+, 61), 208 (33), 163 (20), 137 (100), 107
(26); HRMS (EI) m/z calcd for C23H32O3 356.2351, found
356.2347.
(8R,8′R)-2′-Isopropoxy-4-methoxy-3-lignanol (49). Color-
25
1
less oil; [α]D −26 (c 0.4, CHCl3); H NMR (400 MHz,
CDCl3) δ 0.79 (3H, d, J = 6.5 Hz), 0.85 (3H, d, J = 6.8 Hz),
1.24 (3H, d, J = 5.9 Hz), 1.26 (3H, d, J = 6.0 Hz), 1.75−1.88
(2H, m), 2.30 (1H, dd, J = 13.0, 8.7 Hz), 2.32 (1H, dd, J = 13.5,
8.6 Hz), 2.58 (1H, dd, J = 13.5, 6.4 Hz), 2.72 (1H, dd, J = 13.0,
5.2 Hz), 3.85 (3H, s), 4.48 (1H, m), 5.52 (1H, s), 6.57 (1H, dd,
J = 8.1, 2.0 Hz), 6.69 (1H, d, J = 2.0 Hz), 6.72 (1H, d, J = 8.1
Hz), 6.78−6.82 (2H, m), 7.04 (1H, dd, J = 7.4, 1.7 Hz), 7.11
(1H, ddd, J = 7.8, 7.1, 1.7 Hz); 13C NMR (100 MHz, CDCl3) δ
14.0, 14.3, 22.0, 22.1, 36.0, 36.4, 39.0, 40.7, 56.0, 69.2, 110.3,
112.3, 115.1, 119.6, 120.3, 126.6, 131.00, 131.04, 135.4, 144.5,
145.2, 155.7; MS (EI) m/z 342 (M+, 92), 137 (100), 107 (50);
HRMS (EI) m/z calcd for C22H30O3 342.2195, found
342.2177.
(8R,8′R)-4-Methoxy-2′,3-lignanediol (45). Colorless oil;
25
1
[α]D −30 (c 0.4, CHCl3); H NMR (400 MHz, CDCl3) δ
0.83 (3H, d, J = 6.5 Hz), 0.85 (3H, d, J = 6.6 Hz), 1.79 (1H,
m), 1.87 (1H, m), 2.34 (1H, dd, J = 13.6, 8.7 Hz), 2.42 (1H,
dd, J = 13.6, 8.9 Hz), 2.58 (1H, dd, J = 13.6, 6.2 Hz), 2.67 (1H,
dd, J = 13.6, 5.9 Hz), 3.85 (3H, s), 4.78 (1H, s), 5.58 (1H, s),
6.58 (1H, dd, J = 8.1, 2.1 Hz), 6.70−6.74 (3H, m), 6.83 (1H,
ddd, J = 7.4, 7.3, 0.9 Hz), 7.03 (1H, dd, J = 7.6, 1.5 Hz), 7.07
(1H, d, J = 7.7, 1.7 Hz); 13C NMR (100 MHz, CDCl3) δ 14.1,
35.4, 36.6, 38.6, 40.7, 56.0, 110.4, 115.1, 115.2, 120.4, 120.5,
127.0, 127.5, 131.1, 135.0, 144.5, 145.1, 153.6; MS (EI) m/z
300 (M+, 90), 137 (100), 107 (27); HRMS (EI) m/z calcd for
C19H24O3 300.1725; found 300.1730.
(8R,8′R)-4-Methoxy-2′-methyl-3-lignanol (50). Colorless
25
oil; [α]D −36 (c 0.4, CHCl3); 1H NMR (400 MHz,
CDCl3) δ 0.81 (3H, d, J = 6.6 Hz), 0.87 (3H, d, J = 6.8
Hz), 1.73−1.85 (2H, m), 2.21 (3H, s), 2.35 (1H, dd, J = 13.5,
8.3 Hz), 2.39 (1H, dd, J = 13.5, 9.1 Hz), 2.56 (1H, dd, J = 13.5,
6.6 Hz), 2.65 (1H, dd, J = 13.5, 5.2 Hz), 3.85 (3H, s), 5.53 (1H,
s), 6.57 (1H, dd, J = 8.1, 2.2 Hz), 6.69 (1H, d, J = 2.2 Hz), 6.73
(1H, d, J = 8.1 Hz), 7.03 (1H, m), 7.07−7.12 (3H, m); 13C
NMR (100 MHz, CDCl3) δ 13.7, 14.3, 19.4, 36.5, 38.8, 39.0,
40.6, 56.0, 110.4, 115.1, 120.3, 125.5, 125.7, 129.9, 130.1, 135.0,
136.2, 139.8, 144.6, 145.2; EIMS m/z (%) 298 (M+, 82), 137
(100), 105 (21); HRMS (EI) m/z calcd for C20H26O2
298.1933, found 298.1932.
25
(8R,8′R)-2′,4-Methoxy-3-lignanol (46). Colorless oil; [α]D
−32 (c 0.4, CHCl3); 1H NMR (400 MHz, CDCl3) δ 0.81 (3H,
d, J = 6.2 Hz), 0.82 (3H, d, J = 6.1 Hz), 1.76 (1H, m), 1.84
(1H, m), 2.33 (1H, dd, J = 13.6, 8.6 Hz), 2.39 (1H, dd, J = 13.2,
8.7 Hz), 2.57 (1H, dd, J = 13.6, 6.4 Hz), 2.69 (1H, dd, J = 13.2,
5.8 Hz), 3.74 (3H, s), 3.85 (3H, s), 5.52 (1H, s), 6.57 (1H, dd,
J = 8.2, 2.0 Hz), 6.69 (1H, d, J = 2.0 Hz), 6.73 (1H, d, J = 8.2
Hz), 6.81 (1H, br d, J = 8.1 Hz), 6.85 (1H, ddd, J = 7.6, 7.3, 1.0
Hz), 7.04 (1H, dd, J = 7.3, 1.7 Hz), 7.15 (1H, ddd, J = 7.8, 7.6,
1.7 Hz); 13C NMR (100 MHz, CDCl3) δ 14.06, 14.12, 35.7,
36.3, 38.6, 40.7, 55.1, 56.0, 110.2, 110.3, 115.1, 120.0, 120.4,
126.8, 130.2, 130.6, 135.3, 144.5, 145.2, 157.6; MS (EI) m/z
314 (M+, 100), 137 (88), 121 (45); HRMS (EI) m/z calcd for
C20H26O3 314.1882, found 314.1891.
(8R,8′R)-2′-Fluoro-4-methoxy-3-lignanol (51). Colorless
oil; [α]D −37 (c 0.4, CHCl3); 1H NMR (400 MHz,
25
CDCl3) δ 0.83 (3H, d, J = 6.8 Hz), 0.84 (3H, d, J = 6.7
Hz), 1.77 (1H, m), 1.83 (1H, m), 2.32 (1H, dd, J = 13.5, 8.9
Hz), 2.46 (1H, dd, J = 13.5, 8.9 Hz), 2.58 (1H, dd, J = 13.5, 5.9
Hz), 2.69 (1H, dd, J = 13.5, 6.0 Hz), 3.86 (3H, s), 5.54 (1H, s),
6.57 (1H, dd, J = 8.1, 1.9 Hz), 6.68 (1H, d, J = 1.9 Hz), 6.73
(1H, d, J = 8.1 Hz), 6.97 (1H, br d, J = 8.8 Hz), 7.02 (1H, dd, J
= 7.3, 6.6 Hz), 7.08 (1H, dd, J = 7.4, 1.6 Hz), 7.14 (1H, m); 13C
NMR (100 MHz, CDCl3) δ 13.97, 13.99, 34.1, 37.4, 38.5, 40.7,
56.0, 110.4, 115.09, 115.10 (d, J = 22.3 Hz), 120.3, 123.7 (d, J =
3.7 Hz), 127.3 (d, J = 7.9 Hz), 128.5 (d, J = 15.9 Hz), 131.3 (d,
J = 4.8 Hz), 134.9, 144.6, 145.2, 161.3 (d, J = 244.3 Hz); 19F
NMR (376 MHz, CDCl3) δ −119.1; MS (EI) m/z 302 (M+,
90), 137 (100); HRMS (EI) m/z calcd for C19H23O2F
302.1682, found 302.1680.
(8R,8′R)-2′-Ethoxy-4-methoxy-3-lignanol (47). Colorless
oil; [α]D −28 (c 0.5, CHCl3); 1H NMR (400 MHz,
25
CDCl3) δ 0.80 (3H, d, J = 6.9 Hz), 0.85 (3H, d, J = 6.7
Hz), 1.33 (3H, t, J = 7.0 Hz), 1.75 (1H, m), 1.85 (1H, m), 2.32
(1H, dd, J = 13.6, 8.6 Hz), 2.35 (1H, dd, J = 13.1, 8.7 Hz), 2.58
(1H, dd, J = 13.6, 6.4 Hz), 2.73 (1H, dd, J = 13.1, 5.6 Hz), 3.85
(3H, s), 3.95 (2H, m), 5.51 (1H, s), 6.57 (1H, dd, J = 8.2, 2.1
Hz), 6.68 (1H, d, J = 2.1 Hz), 6.72 (1H, d, J = 8.2 Hz), 6.79
(1H, d, J = 8.0 Hz), 6.83 (1H, dd, J = 7.4, 7.4 Hz), 7.04 (1H,
dd, J = 7.4, 1.7 Hz), 7.13 (1H, ddd, J = 7.4, 7.4, 1.7 Hz); 13C
NMR (100 MHz, CDCl3) δ 14.2, 14.3, 14.9, 35.8, 36.4, 38.8,
40.6, 56.0, 63.2, 110.3, 111.0, 115.1, 119.9, 120.3, 126.7, 130.3,
130.8, 135.4, 144.5, 145.2, 157.0; MS (EI) m/z 328 (M+, 78),
137 (100), 135 (54); HRMS (EI) m/z calcd for C21H28O3
328.2038, found 328.2020.
(8R,8′R)-4-Methoxy-3,3′-lignanediol (52). Colorless oil;
25
1
[α]D −19 (c 0.5, CHCl3); H NMR (400 MHz, CDCl3) δ
0.80 (3H, d, J = 6.7 Hz), 0.81 (3H, d, J = 6.7 Hz), 1.73−1.81
(2H, m), 2.36 (1H, dd, J = 13.5, 8.5 Hz), 2.38 (1H, dd, J = 13.5,
8.6 Hz), 2.52 (1H, dd, J = 13.4, 6.5 Hz), 2.57 (1H, dd, J = 13.4,
6.4 Hz), 3.86 (3H, s), 4.90−5.40 (1H, br), 5.65 (1H, br s), 6.54
(1H, dd, J = 2.1, 1.7 Hz), 6.58 (1H, dd, J = 8.2, 2.1 Hz), 6.64−
6.69 (3H, m), 6.74 (1H, d, J = 8.2 Hz), 7.11 (1H, dd, J = 7.8,
7.8 Hz); 13C NMR (100 MHz, CDCl3) δ 13.8, 13.9, 37.1, 37.6,
40.6, 41.1, 56.0, 110.4, 112.7, 115.1, 115.8, 120.6, 121.6, 129.2,
134.9, 143.5, 144.6, 145.0, 155.4; MS (EI) m/z 300 (M+, 91),
137 (100); HRMS (EI) m/z calcd for C19H24O3 300.1725,
found 300.1733.
(8R,8′R)-2′-Butoxy-4-methoxy-3-lignanol (48). Colorless
25
oil; [α]D −17 (c 0.5, CHCl3); 1H NMR (400 MHz,
CDCl3) δ 0.80 (3H, d, J = 6.8 Hz), 0.84 (3H, d, J = 6.8
Hz), 0.97 (3H, t, J = 7.4 Hz), 1.48 (2H, m), 1.71 (2H, m), 1.76
(1H, m), 1.86 (1H, m), 2.30 (1H, dd, J = 13.5, 8.9 Hz), 2.36
(1H, dd, J = 13.0, 8.9 Hz), 2.59 (1H, dd, J = 13.5, 6.1 Hz), 2.72
(1H, dd, J = 13.0, 5.6 Hz), 3.86 (3H, s), 3.89 (2H, m), 5.51
(1H, s), 6.57 (1H, dd, J = 8.1, 2.0 Hz), 6.68 (1H, d, J = 2.0 Hz),
6.72 (1H, d, J = 8.1 Hz), 6.79 (1H, d, J = 8.2 Hz), 6.84 (1H, d, J
= 7.3 Hz), 7.04 (1H, dd, J = 7.3, 1.4 Hz), 7.13 (1H, ddd, J = 7.8,
7.7, 1.4 Hz); 13C NMR (100 MHz, CDCl3) δ 13.9, 14.1, 19.4,
5309
dx.doi.org/10.1021/jf5010572 | J. Agric. Food Chem. 2014, 62, 5305−5315

Journal of Agricultural and Food Chemistry
Article
Table 1. Cytotoxic Activities of (8R,8′R)-Secoisolariciresinol (1), (8R,8′R)-9,9′-Anhydrosecoisolariciresinol (2), All
Stereoisomers of Dihydroguaiaretic Acid (3−5), and 9-,9′-Derivatives of (8R,8′R)-Dihydroguaiaretic Acid 6−15 against HL-60
and HeLa Cells (IC50 SD, n = 3)
25
(8R,8′R)-3′,4-Methoxy-3-lignanol (53). Colorless oil; [α]D
−25 (c 0.6, CHCl3); 1H NMR (400 MHz, CDCl3) δ 0.82 (6H,
d, J = 6.8 Hz), 1.73−1.84 (2H, m), 2.33 (1H, dd, J = 13.6, 8.5
Hz), 2.39 (1H, dd, J = 13.4, 8.6 Hz), 2.56 (1H, dd, J = 13.6, 6.3
Hz), 2.62 (1H, dd, J = 13.4, 6.2 Hz), 3.78 (3H, s), 3.86 (3H, s),
5.55 (1H, s), 6.56 (1H, dd, J = 8.2, 1.9 Hz), 6.65 (1H, m),
6.68−6.74 (4H, m), 7.16 (1H, dd, J = 7.9, 7.8 Hz); 13C NMR
(100 MHz, CDCl3) δ 14.0, 38.0, 38.2, 40.7, 41.5, 55.1, 56.0,
110.4, 110.9, 114.6, 115.1, 120.3, 121.5, 129.0, 135.0, 143.3,
144.6, 145.2, 159.4; MS (EI) m/z 314 (M+, 100), 137 (96), 122
(37); HRMS (EI) m/z calcd for C20H26O3 314.1882, found
314.1890.
CDCl3) δ 0.815 (3H, d, J = 6.8 Hz), 0.821 (3H, d, J = 6.7
H), 1.72−1.83 (2H, m), 2.34 (1H, dd, J = 13.5, 8.5 Hz), 2.41
(1H, dd, J = 13.5, 8.6 Hz), 2.55 (1H, dd, J = 13.5, 6.4 Hz), 2.64
(1H, dd, J = 13.5, 6.3 Hz), 3.87 (3H, s), 5.54 (1H, s), 6.56 (1H,
d, J = 8.1, 2.1 Hz), 6.68 (1H, d, J = 2.1 Hz), 6.74 (1H, d, J = 8.1
Hz), 6.79 (1H, ddd, J = 10.0, 2.1, 1.7 Hz), 6.83−6.89 (2H, m),
7.20 (1H, ddd, J = 7.8, 7.7, 6.3 Hz): 13C NMR (100 MHz,
CDCl3) δ 13.9, 14.0, 37.9, 38.2, 40.7, 41.2, 56.0, 110.4, 112.5
(d, J = 20.8 Hz), 115.1, 115.7 (d, J = 21.0 Hz), 120.3, 124.7,
129.4 (d, J = 8.5 Hz), 134.8, 144.3 (d, J = 7.1 Hz), 144.6, 145.3,
162.8 (d, J = 245.3 Hz); 19F NMR (376 MHz, CDCl3) δ
−115.2; MS (EI) m/z 302 (M+, 74), 137 (100); HRMS (EI)
m/z calcd for C19H23FO2 302.1682, found 302.1682.
(8R,8′R)-4-Methoxy-3′-methyl-3-lignanol (54). Colorless
25
oil; [α]D −26 (c 0.6, CHCl3); 1H NMR (400 MHz,
(8R,8′R)-4-Methoxy-3,4′-lignanediol (56). Colorless oil;
25
1
CDCl3) δ 0.81 (3H, d, J = 6.6 Hz), 0.82 (3H, d, J = 6.7
Hz), 1.73−1.84 (2H, m), 2.30−2.35 (1H, overlapped), 2.31
(3H, s), 2.37 (1H, dd, J = 13.4, 8.6 Hz), 2.57 (1H, dd, J = 13.4,
6.0 Hz), 2.61 (1H, dd, J = 13.4, 6.0 Hz), 3.86 (3H, s), 5.54 (1H,
s), 6.57 (1H, dd, J = 8.2, 1.9 Hz), 6.69 (1H, d, J = 1.9 Hz), 6.73
(1H, d, J = 8.2 Hz), 6.90 (1H, br d, J = 7.2 Hz), 6.91 (1H, br s),
[α]D −21 (c 0.5, CHCl3); H NMR (CDCl3) δ 0.797 (3H,
d, J = 6.5 Hz), 0.801 (3H, d, J = 6.7 Hz), 1.70−1.78 (2H, m),
2.32 (1H, dd, J = 13.5, 8.9 Hz), 2.35 (1H, dd, J = 13.5, 8.9 Hz),
2.55 (1H, dd, J = 13.5, 6.7 Hz), 2.56 (1H, dd, J = 13.5, 6.7 Hz),
3.86 (3H, s), 4.77 (1H, s), 5.56 (1H, s), 6.57 (1H, dd, J = 8.1,
2.0 Hz), 6.68 (1H, d, J = 2.0 Hz), 6.72 (2H, d, J = 8.5 Hz), 6.74
(1H, d, J = 8.1 Hz), 6.95 (2H, d, J = 8.5 Hz); 13C NMR (100
MHz, CDCl3) δ 13.9, 38.1, 38.3, 40.5, 40.8, 56.0, 110.4, 115.0,
115.1, 120.4, 130.0, 133.9, 135.0, 144.5, 145.2, 153.4; MS (EI)
m/z 300 (M+, 95), 137 (100), 107 (35); HRMS (EI) m/z calcd
for C19H24O3 300.1725, found 300.1731.
6.98 (1H, br d, J = 7.5 Hz), 7.14 (1H, dd, J = 7.5, 7.1 Hz); 13
C
NMR (100 MHz, CDCl3) δ 14.0, 21.4, 38.0, 38.3, 40.7, 41.3,
56.0, 110.3, 115.1, 120.3, 126.0, 126.3, 128.0, 129.8, 135.0,
137.6, 141.6, 144.5, 145.2; MS (EI) m/z 298 (M+, 25), 137
(100); HRMS (EI) m/z calcd for C20H26O2 298.1933, found
298.1929.
25
(8R,8′R)-4,4′-Methoxy-3-lignanol (57). Colorless oil; [α]D
(8R,8′R)-3′-Fluoro-4-methoxy-3-lignanol (55). Colorless
−22 (c 0.8, CHCl3); 1H NMR (400 MHz, CDCl3) δ 0.80 (3H,
d, J = 6.7 Hz), 0.81 (3H, d, J = 6.5 Hz), 1.75 (2H, m), 2.32
25
oil; [α]D −26 (c 0.5, CHCl3); 1H NMR (400 MHz,
5310
dx.doi.org/10.1021/jf5010572 | J. Agric. Food Chem. 2014, 62, 5305−5315

Journal of Agricultural and Food Chemistry
Article
Table 2. Cytotoxic Activities of 7-Aryl Derivatives of (8R,8′R)-Dihydroguaiaretic Acid against HL-60 and HeLa Cells (IC50
SD, n = 3)
no.
R
HL-60 IC50 (μM) SD HeLa IC50 (μM) SD no.
R
HL-60 IC50 (μM) SD HeLa IC50 (μM) SD
16
17
18
19
20
21
22
23
24
25
26
27
28
29
H
10.3 2.4
9.6 2.7
6.3 0.2
7.2 2.4
5.9 1.5
13.5 0.8
10.0 0.5
26.1 3.4
27.1 3.3
7.5 2.2
15.9 2.9
11.0 0.1
9.1 2.6
5.9 0.5
31.8 1.5
9.3 2.0
30
31
32
33
34
35
36
37
38
39
40
41
42
43
4-OCH3
12.5 1.5
6.6 1.9
16.1 2.0
13.4 2.5
10.6 2.5
5.9 0.2
10.8 0.6
31.9 3.5
8.7 1.2
7.3 1.9
28.6 1.9
9.0 3.6
12.6 1.0
28.9 1.7
27.6 2.6
10.7 1.1
17.5 2.5
19.1 0.3
18.3 0.4
13.6 1.1
7.6 1.0
2-OH
4-O(CH2)3CH3
4-CH3
2-OCH3
2-O(CH2)3CH3
2-CH3
24.4 0.5
9.3 0.3
4-F
17.3 0.4
18.7 0.3
18.5 0.7
26.5 2.1
24.3 0.4
10.6 1.4
18.7 0.4
17.9 1.1
17.0 1.8
10.8 0.8
4-Cl
2-F
3,4-OH
2-Cl
3,5-OH
3-OH
3,4-OCH3
3-OH-4-OCH3
3-OCH2CH3-4-OH
3,4-OCH2O
3,5-OCH3-4-OH
3,4,5-OH
30.5 1.9
7.5 0.5
3-OCH3
3-O(CH2)3CH3
3-CH3
27.9 2.8
32.7 2.5
6.8 0.5
3-F
3-Cl
34.9 2.4
>100 (36% inhibition)
4-OH
(1H, dd, J = 13.6, 8.5 Hz), 2.36 (1H, dd, J = 13.6, 8.5 Hz), 2.55
(1H, dd, J = 13.6, 5.9 Hz), 2.59 (1H, dd, J = 13.6, 5.9 Hz), 3.79
(3H, s), 3.86 (3H, s), 5.55 (1H, s), 6.57 (1H, dd, J = 8.1, 2.0
Hz), 6.69 (1H, d, J = 2.0 Hz), 6.73 (1H, d, J = 8.1 Hz), 6.80
(2H, d, J = 8.6 Hz), 7.01 (2H, d, J = 8.6 Hz); 13C NMR (100
MHz, CDCl3) δ 13.9, 38.2, 38.3, 40.5, 40.8, 55.2, 56.0, 110.4,
113.5, 115.1, 120.3, 129.8, 133.7, 135.0, 144.5, 145.2, 157.6; MS
(EI) m/z 314 (M+, 89), 137 (75), 121 (100), 101 (28); HRMS
(EI) m/z calcd for C20H26O3 314.1882, found 314.1885.
(8R,8′R)-4-Methoxy-4′-methyl-3-lignanol (58). Colorless
Cytotoxic Assay. Human cervical carcinoma cell line HeLa
cells and human promyelocytic leukemia cell line HL-60 cells
were obtained from American Type Culture Collection
(Manassas, VA). Colon 26 colon adenocarcinoma cells derived
from BALB/c mouse were kindly provided by the Institute of
Development, Aging and Cancer, Tohoku University (Sendai,
Japan). Vero cells (JCRB0111) were purchased from JCRB Cell
Banks. These cells were cultured in the DMEM medium
(Sigma, St. Louis, MO) supplemented with 10% fetal bovine
serum (FBS) at 37 °C in a humidified atmosphere of 5% CO2.
HeLa, Colon 26, and Vero cells were inoculated in 96-well
culture plates at 2.0 × 104 cells/well suspended in the DMEM
medium, and HL-60 cells were inoculated in 96-well culture
plates at 1.0 × 104 cells/well suspended in RPMI1640 with FBS
and various concentrations of different compounds. After
cultivation for 48 h, the cell viability was assessed by a WST-8
reduction assay (Dojin Laboratories, Japan). The WST-8
reduction activity of the cells is presented as the ratio of cell
activity. Briefly, a WST-8 solution was added to the culture
medium at 10% concentration and incubated for 1 h at 37 °C
prior to colorimetry at 450 nm. The single cytotoxic assay was
performed in triplicate.
oil; [α]D −23 (c 0.5, CHCl3); 1H NMR (400 MHz,
25
CDCl3) δ 0.80 (3H, d, J = 6.7 Hz), 0.81 (3H, d, J = 6.7
Hz), 1.77 (2H, m), 2.29−2.35 (1H, overlapped), 2.31 (3H, s),
2.37 (1H, dd, J = 13.3, 8.5 Hz), 2.56 (1H, dd, J = 13.3, 5.9 Hz),
2.61 (1H, dd, J = 13.3, 5.7 Hz), 3.85 (3H, s), 5.55 (1H, s), 6.57
(1H, dd, J = 8.2, 1.9 Hz), 6.69 (1H, d, J = 1.9 Hz), 6.73 (1H, d,
J = 8.2 Hz), 6.99 (2H, d, J = 7.9 Hz), 7.06 (2H, d, J = 7.9 Hz);
13C NMR (100 MHz, CDCl3) δ 13.9, 21.0, 38.2, 38.3, 40.8,
41.0, 56.0, 110.3, 115.1, 120.3, 128.8, 128.9, 134.9, 135.0, 138.5,
144.5, 145.2; MS (EI) m/z 298 (M+, 58), 137 (100), 105 (41);
HRMS (EI) m/z calcd for C20H26O2 298.1933, found
298.1930.
(8R,8′R)-4′-Fluoro-4-methoxy-3-lignanol (59). Colorless
25
oil; [α]D −26 (c 0.6, CHCl3); 1H NMR (400 MHz,
RESULTS AND DISCUSSION
■
CDCl3) δ 0.80 (3H, d, J = 6.7 Hz), 0.81 (3H, d, J = 6.5
Hz), 1.70−1.78 (2H, m), 2.33 (1H, dd, J = 13.5, 8.3 Hz), 2.38
(1H, dd, J = 13.5, 8.4 Hz), 2.54 (1H, dd, J = 13.5, 6.1 Hz), 2.60
(1H, dd, J = 13.5, 6.1 Hz), 3.86 (3H, s), 5.54 (1H, s), 6.55 (1H,
dd, J = 8.2, 2.0 Hz), 6.68 (1H, d, J = 2.0 Hz), 6.73 (1H, d, J =
8.2 Hz), 6.93 (2H, m), 7.03 (2H, m); 13C NMR (100 MHz,
CDCl3) δ 13.8, 13.9, 38.1, 38.2, 40.6, 40.7, 56.0, 110.4, 114.8
(d, J = 21.1 Hz), 115.1, 120.3, 130.2 (d, J = 7.8 Hz), 134.9,
137.2 (d, J = 3.0 Hz), 144.6, 145.2, 161.1 (d, J = 242.6); 19F
NMR (376 MHz, CDCl3) δ −119.1; MS (EI) m/z 302 (M+,
92), 137 (100); HRMS (EI) m/z calcd for C19H23FO2
302.1682, found 302.1679.
Syntheses of Derivatives. As shown in Scheme 1, 9- and
9′-derivatives of (8R,8′R)-DGA 6−15 were synthesized from
dibenzyloxymatairesinol 61. The aromatic derivatives 16−59
were synthesized by the previously described method15 with
modification.
Cytotoxic Activities of 9- and 9′-Derivatives against
HL-60 and HeLa Cells. Table 1 shows the cytotoxic activity of
dietary (8R,8′R)-secoisolariciresinol (SECO) (1) and its 9 and
9′-derivatives against HL-60 and HeLa cells. (8R,8′R)-SECO
(1) and (8R,8′R)-9,9′-anhydrosecoisolariciresinol (2), which is
a metabolite from (−)-(8R,8′R)-SECO, did not show the
remarkable activity at 100 μM, suggesting that the oxidations at
5311
dx.doi.org/10.1021/jf5010572 | J. Agric. Food Chem. 2014, 62, 5305−5315

Journal of Agricultural and Food Chemistry
Article
were reduced. All stereoisomers of dihydroguaiaretic acids
exhibited the almost same level of activities (IC50 = 22−41
μM). The stereochemistry of 8 and 8′-positions did not affect
the activity. The activity of 9, 9′-acetoxy derivative 6 was 2−3-
fold less potent than DGA stereoisomers, and the activity of 9,
9′-nitrile derivative 8 was very weak (IC50 > 100 μM); however,
a similar level of activity to DGA stereoisomers was observed in
9, 9′-methoxy derivative 7. Compared with inactive tetrahy-
drofuran structure 2, the disadvantage of the ring structure was
suggested. Even if the oxygen atoms are present at 9- and 9′-
positions, the methoxy derivative kept the activity. For further
research on the 9-position, the activities of 9-derivatives of
(8R,8′R)-DGA 9−15 were tested. Compared with inactive
(−)-(8R,8′R)- SECO (1), 9′-dehydroxy SECO 9 showed the
weak activity (IC50 = 79−98 μM). The activities of 9-acetoxy
and 9-methoxy DGA derivative 10 and 11 were the same level
as those of all stereoisomers of DGA 3−5, 9,9′-acetoxy 6, and
9,9′-methoxy 7 derivative. On the other hand, 9-alkyl DGA
derivative 13 and 14 displayed 3−4-fold higher activity than
that of (8R,8′R)-DGA (3). The activity level of 9-butyl DGA
13 and 9-isopropyl DGA 14 was similar, indicating the
importance of higher hydrophobic structure for the higher
activity. Because 9-fluoro derivative 12 showed comparable
activity to that of (8R,8′R)-DGA (3), the activity would not be
affected by an electron-withdrawing atom. The activity of basic
derivative, 9-dimethylamino DGA derivative 15, was very low.
The relationship between cytotoxic activity and 9-, 9′-structure
of butane-type lignan containing dietary secoisolariciresinol
became clear. The higher hydrophobic compounds exhibited
higher activity.
Table 3. Cytotoxic Activities of (8R,8′R)-7-(3-Hydroxy-4-
methoxyphenyl)-7′-aryl Dihydroguaiaretic Acid Derivatives
against HL-60 and HeLa Cells (IC50 SD, n = 3)
no.
R
HL-60 IC50 (μM) SD HeLa IC50 (μM) SD
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
H
8.2 1.8
3.5 0.6
1.6 0.2
0.8 0.4
7.2 0.4
3.8 0.3
2.7 0.04
3.7 0.8
8.7 0.6
7.1 1.2
5.9 0.3
9.5 0.6
4.3 0.4
4.5 1.2
5.3 1.3
5.8 0.7
9.5 1.3
3.8 1.1
2.6 0.6
1.7 0.6
6.7 1.2
4.3 1.0
4.7 1.3
4.8 0.6
9.7 0.7
6.2 1.0
5.1 0.5
9.5 0.2
9.8 1.3
5.4 1.5
5.9 1.1
9.9 0.3
2′-OH
2′-OCH3
2′-OCH2CH3
2′-O(CH2)3CH3
2′-OCH(CH3)2
2′-CH3
2-F
3′-OH
3′-OCH3
3′-CH3
3′-F
4′-OH
4′-OCH3
4′-CH3
4′-F
9- and 9′-positions on the butane chain are disadvantageous for
the activity. This fact was ascertained by the examination using
dihydroguaiaretic acids (DGA) 3−5, whose 9- and 9′-positions
Cytotoxic Activities of 7-Aryl Derivatives against HL-
60 and HeLa Cells. As a next step, the effect of the substituent
Table 4. Cytotoxic Activities of 3, 38, and 47 against Colon 26 and Vero Cells (IC50 SD, n = 3)
5312
dx.doi.org/10.1021/jf5010572 | J. Agric. Food Chem. 2014, 62, 5305−5315

Journal of Agricultural and Food Chemistry
Article
of the aromatic ring on the activity was examined. Table 2
shows the activities of 7-derivatives of (8R,8′R)-DGA. Without
a substituent on the 7-aromatic ring, derivative 16 showed 2.6-
fold higher activity than (8R,8′R)-DGA (3) against HL-60 cells
and similar activity to 3 against HeLa cells. It could be assumed
that the 3-methoxy group and 4-hydroxy group of 3 do not play
an important role to keep 10−30 μM of IC50 values. As for 2-
substituted derivatives 17−22, all derivatives showed 2−4-fold
higher activity than that of (8R,8′R)-DGA (3) against HL-60
cells. The activities of 2-methoxy derivative 18, 2-butoxy
derivative 19, and 2-methyl derivative 20 bearing an electron
donor group were about 2-fold higher than those of hydrophilic
2-hydroxy derivative 17 and 2-halo derivative 21 and 22 bearing
an electron-withdrawing group, suggesting that the hydro-
phobic and electron donor substituents on the 2-position are
favorable for higher activity against HL-60 cells. Because the
activity of 2-butoxy derivative 19 was similar to that of 2-
methoxy derivative 18 against HL-60 cells, the length of alkoxy
group would not affect the activity. Against HeLa cells,
hydrophilic 2-hydroxy derivative 17 and 2-butoxy derivative
19 bearing a longer alkoxy group exhibited 2-fold higher activity
than that of (8R,8′R)-DGA (3). On the other hand, the same
level of activities as that of (8R,8′R)-DGA (3) were observed in
2-methoxy derivative 18, 2-methyl derivative 20, and 2-halo
derivative 21 and 22. The 2-hydroxy derivative 17 and 2-butoxy
derivative 19 showed higher activity than (8R,8′R)-DGA (3)
against both cells. The effect of 3-substituent was also
examined. It was shown that the activities of 3-hydroxy
derivative 23 and 3-methoxy derivative 24 were similar to that
of (8R,8′R)-DGA (3) against both HL-60 and HeLa cells. The
more hydrophobic derivatives, 3-butoxy derivative 25, 3-methyl
derivative 26, 3-fluoro derivative 27, and 3-chloro derivative 28
were 2−3-fold more effective than (8R,8′R)-DGA (3) against
HL-60 cells. Against HeLa cells, the activity level of 3-methyl
derivative 26, 3-fluoro derivative 27, and 3-chloro derivative 28
was almost the same as that of (8R,8′R)-DGA (3); however, 3-
butoxy derivative 25 exhibited 2-fold more potent activity than
(8R,8′R)-DGA (3). The 3-butoxy derivative 25 bearing a
longer and hydrophobic group displayed the highest activity
among the 3-substituted derivatives against both cells. In the
case of the 4-position, 4-hydroxy derivative 29 and 4-butoxy
derivative 31 were 4-fold more potent and 4-methoxy derivative
30 and 4-halide derivative 33 and 34 were 2-fold more potent
than that of (8R,8′R)-DGA (3) against HL-60 cells. Against
HeLa cells, 2-fold higher activities than that of (8R,8′R)-DGA
(3) were observed in 4-hydroxy derivative 29 and 4-butoxy
derivative 31. The activities of 4-methyl derivative 32 against
HL-60 and HeLa cells and 4-methoxy derivative 30 and 4-
halide derivative 33 and 34 against HeLa cells were similar to
that of (8R,8′R)-DGA (3). The phenolic derivative and longer
alkoxy derivative were more effective at the 4-position as well as
the 2-position against both cells. The activities of all
monosubstituted derivatives are not less potent than that of
(8R,8′R)-DGA (3). Especially, the IC50 values of 2-hydroxy
derivative 17 and 2-butoxy derivative 19 were less than 10 μM
against both HL-60 and HeLa cells. The activities of
disubstituted compounds 35−40 were also compared. The
3,4-dihydroxy derivative 35 was 5-fold more potent against HL-
60 cells and 2-fold more potent against HeLa cells than 3,4-
dimethoxy derivative 37 and 3,4-methylenedioxy derivative 40,
whose activities were similar to that of (8R,8′R)-DGA (3). The
3,5-dihydroxy derivative 36 showed 3-fold and 4-fold higher
activity against HL-60 cells and HeLa cells, respectively, than
3,4-dimethoxy derivative 37. These facts mean that the
hydrophilic disubstituted compound is advantageous for higher
activity. The longer-chain derivative, 3-ethoxy-4-hydroxy
derivative 39 enhanced the activity, showing 4-fold higher
activity than (8R,8′R)-DGA (3) against HL-60 cells; however,
similar activity to (8R,8′R)-DGA (3) was observed against
HeLa cells. It is noteworthy that 3-hydroxy-4-methoxy
derivative 38, in which the positions of substituents are
exchanged from that of natural DGA, showed 3-fold more
potent activity than (8R,8′R)-DGA (3) against both cells. The
positions of the hydrophobic and hydrophilic group affected
the activity. The addition of a methoxy group to the 5-position
of (8R,8′R)-DGA (3) enhanced the activity. The activity of 3,5-
methoxy-4-hydroxy derivative 41 was 3-fold more potent than
(8R,8′R)-DGA (3) against both cells. Compared with HeLa
cells, HL-60 cells were more sensitive to more hydrophilic
DGA derivatives. The activity of 3,4,5-hydroxy derivative 42
and pentahydroxy derivative 43 was 2-fold higher and
comparable to (8R,8′R)-DGA (3), respectively. On the other
hand, against HeLa cells, the activities of 3,4,5-hydroxy
derivative 42 and pentahydroxy derivative 43 were similar to
and much lower than (8R,8′R)-DGA (3), respectively. The
position and the number of hydrophobic and hydrophilic
groups affected the activity. The 7-(3-hydroxy-4-methoxy)-7′-
(3-methoxy-4- hydroxy) derivative 38 showed less than 9 μM
IC50 value against both cells.
Cytotoxic Activities of 7′-Aryl Derivatives against HL-
60 and HeLa Cells. With the results of the cytotoxic
experiment on 7-aryl derivatives, we selected 3-hydroxy-4-
methoxyphenyl group as an aryl group for 7-position to detect
the effect of the 7′-aryl group on the activity. The cytotoxic
activities of 7-(3-hydroxy-4-methoxyphenyl)-7′-aryl derivatives
44−59 were evaluated. The activity of 7′-phenyl derivative 44
was comparable to that of 7′-(3-methoxy-4-hydroxyphenyl)
derivative 38. The presence of substituent on 7′-aryl group is
not necessary to exhibit the activity level of 38. Introduction of
the substituent to the 2′-position of 44 enhanced the activity
against both cells. The 2′-hydroxy derivative 45 and 2′-F
derivative 51 showed 2-fold higher activity, and 2′-methoxy
derivative 46 and 2′-methyl derivative 50 were 5-fold and 3-fold
more potent, respectively, than 38 against HL-60 cells. These
facts mean that the electron donor substituent is more effective
than the hydrophilic and electron-withdrawing substituent on
the 2′-position. The activity of 2′-ethoxy derivative 47 was 2-
fold potent than 2′-methoxy derivative, showing the highest
activity among 2′-derivatives; however, the longer and bulky
substituent at the 2′-position decreased activity. The butoxy 48
and isopropoxy 49 derivative exhibited 2−4-fold less potent
activity than that of the 2′-methoxy derivative. Against HeLa
cells, 2′-ethoxy derivative 47 was also most effective among the
2′-derivatives, showing 4-fold higher activity than 38. The
shorter alkoxy derivative 46 and longer and bulker alkoxy
derivative 48 and 49 showed weaker activity than that of 2′-
ethoxy derivative 47 against HeLa cells as well as against HL-60
cells. The activities of 2′-methyl derivative 50, 2′-fluoro
derivative 51, and 2′-hydroxy derivative 45 were 1.6−2-fold
higher than that of 38 against HeLa cells. The 3′- and 4′-
derivatives 52−59 showed comparable activity to 38 against
both cells except for 4′-hydroxy derivative 56 and 4′-methoxy
derivative 57, which were 1.6-fold more potent than 38 against
HL-60 cells. The presence of substituent on 2′-position of 7-(3-
hydroxy-4-methoxyphenyl)-7′-aryl derivatives was important
for the higher activity. 7-(3-Hydroxy-4-methoxyphenyl)-7′-(2′-
5313
dx.doi.org/10.1021/jf5010572 | J. Agric. Food Chem. 2014, 62, 5305−5315

Journal of Agricultural and Food Chemistry
Article
Funding
ethoxyphenyl) derivative 47 showed the most potent activity
against both cells in this research. The shorter and longer
alkoxy group at the 2′-position decreased activity (Table 3).
Cytotoxic Activity against Colon 26 and Vero Cells.
Finally, the natural (8R,8′R)-DGA (3) and the higher active
derivatives 38 and 47 against HL-60 and HeLa cells were
applied to cytotoxic assay against colon 26 as colorectal cancer
cells and Vero cells as normal cells (Table 4). Although the
activities against colon 26 cells were weaker than against HL-60
and HeLa cells, these derivatives were also effective. Especially,
the IC50 value of compounds 47 against colon 26 was less than
10 μM. Theses derivatives were also effective against Vero cells;
however, their activities were weaker than against HL-60 and
HeLa cells. The activity of derivative 38 was 3-fold less active,
and derivative 47 showed 4−8.5-fold less activity against Vero
cells than against HL-60 and HeLa cells, respectively. The most
specific activity against cancer cells was observed in derivative
47.
In conclusion, we clarified the structure-cytotoxic activity
relationship of butane-type lignan containing dietary lignan,
secoisolariciresinol, for the first time. The hydroxy groups on 9-
and 9′-positions of secoisolariciresinol are disadvantageous for
the activity. This fact was confirmed by the appearance of
activity of 9- and 9′-reductive type lignan, dihydroguaiaretic
acid (DGA), whose all stereoisomers showed same level of
activities. In the course of the evaluation of activities of the
derivatives, 9-butyl (8R,8′R)-DGA derivative 13, 7-(2-hydrox-
yphenyl) (8R,8′R)-DGA derivative 17, 7-(2-butoxyphenyl)
(8R,8′R)- DGA derivative 19, 7-(3,5-dimethoxy-4-hydroxy
(8R,8′R)-DGA derivative 41, and 7-(3-hydroxy-4-methoxy-
phenyl) (8R,8′R)-DGA derivative 38 showed 2−4-fold higher
activity than (8R,8′R)-DGA (3). Finally, 7-(3-hydroxy-4-
methoxyphenyl)-7′-(2′-ethoxyphenyl) DGA derivative 47
showed highest cytotoxic activity in this research, showing
We are grateful to Marutomo Co. for financial support.
Notes
The authors declare no competing financial interest.
ACKNOWLEDGMENTS
■
Part of this study was performed at INCS (Johoku station) of
Ehime University. Our thanks to the president of Ehime
University for supporting this project.
REFERENCES
■
(1) Penalvo, J. L.; Adlercreutz, H.; Uehara, M.; Ristimaki, A.;
Watanabe, S. Lignan content of selected foods from Japan. J. Agric.
Food Chem. 2008, 56, 401−409.
(2) Ayres, D. C.; Loike, J. D. Biological and clinical properties of
podophyllotoxin and other lignans. Lignans; Cambridge University
Press: Cambridge, U.K., 1990; pp 85−112.
(3) Osawa, T. Phenolic antioxidants on dietary plants as
antimutagens. In Phenolic compounds in food and their effects on health
II, antioxidants and cancer prevention; Huang, M.-T, Ho, C.-T, Lee, C.
Y., Eds.; American Chemical Society: Washington, DC, 1992; pp 135−
149.
(4) Kawahara, S.; Iwata, I.; Fujita, E.; Yamawaki, M.; Nishiwaki, H.;
Sugahara, T.; Yamauchi, S.; Akiyama, K.; Kishida, T. IgE-suppressive
activity of (−)-matairesinol and its structure-activity relationship.
Biosci. Biotechnol. Biochem. 2010, 74, 1878−1883.
(5) Yamawaki, M.; Nishi, K.; Nishimoto, S.; Yamauchi, S.; Akiyama,
K.; Kishida, T.; Maruyama, M.; Nishiwaki, H.; Sugahara, T.
Immunomodulatory effect of (−)-matairesinol in vivo and ex vivo.
Biosci. Biotechnol. Biochem. 2011, 75, 859−863.
(6) Tominaga, S.; Sugahara, T.; Nishimoto, S.; Yamawaki, M.;
Nakashima, Y.; Kishida, T.; Akiyama, K.; Maruyama, M.; Yamauchi, S.
Biosci. Biotechnol. Biochem. 2009, 73, 35−39.
(7) Yamauchi, S.; Kishida, T.; Sugahara, T.; Yamawaki, M.;
Nishimoto, S.; Shinomiya, Y.; Yamamoto, T. Inhibition of the
discoloration of yellowtail dark muscle by lignan. Biosci. Biotechnol.
Biochem. 2009, 73, 1718−1721.
(8) Yamauchi, S.; Kawahara, S.; Wukirsari, T.; Nishiwaki, H.; Nishi,
K.; Sugahara, T.; Akiyama, K.; Kishida, T. Structure−cytotoxic activity
relationship of sesquilignan, morinol A. Bioorg. Med. Chem. Lett. 2013,
23, 4923−4930.
(9) Niemeyer, H. B.; Honig, D. M.; Kulling, S. E.; Metzler, M.
Studies on the metabolism of the plant lignans secoisolariciresinol and
matairesinol. J. Agric. Food Chem. 2003, 51, 6317−6325.
(10) Struijs, K.; Vincken, J.-P; Gruppen, H. Bacterial conversion of
secoisolariciresinol and anhydrosecoisolariciresinol. J. Appl. Microbiol.
2009, 107, 308−317.
(11) Ho, S. S. H.; Go, M. L. Restraining the flexibility of the central
linker in terameprocol results in constrained analogs with improved
growth inhibitory activity. Bioorg. Med. Chem. Lett. 2013, 23, 6127−
6133.
32-fold and 13-fold higher activity against HL-60 cells (IC50
=
0.8 μM) and HeLa cells (IC50 = 1.7 μM), respectively, than
(8R,8′R)-DGA (3). This derivative 47 was also effective against
colon 26 cells. Although (8R,8′R)-9,9′-anhydrosecoisolaricir-
esinol (2), which is a candidate for metabolite of dietary
secoisolariciresinol, did not show the cytotoxic activity in this
research, the higher cytotoxic active dihydroguaiaretic acid
(DGA) derivative was prepared as a new lead compound,
whose main chemical structure is same as that of dietary
secoisolariciresinol. Since there is a possibility that dietary
lignan derivatives will be detected as a metabolite, the study on
structure−activity relationship of lignan should be promoted to
examine the effect of dietary lignan on health and safety. This
result would contribute to development of newer lead
compounds of medicine based on natural dietary compounds.
(12) Mansoor, T. A.; Ramalho, R. M.; Rodrigues, C. M. P.; Ferreira,
M.-J. U. Dibenzylbutane- and butyrolactone-type lignans as apoptosis
inducers in human hepatoma HuH-7 cells. Phytother. Res. 2012, 26,
692−696.
(13) Yamauchi, S.; Sugahara, T.; Matsugi, J.; Someya, T.; Masuda, T.;
Kishida, T.; Akiyama, K.; Maruyama, M. Effect of the benzylic
structure of lignan on antioxidant activity. Biosci. Biotechnol. Biochem.
2007, 71, 2283−2290.
ASSOCIATED CONTENT
■
S
* Supporting Information
Supplementary data includes melting points and both NMR
and MS data for investigated compounds. Supporting
Information also includes additional details for compound
syntheses. This material is available free of charge via the
Internet at http://pubs.acs.org.
(14) Yamauchi, S.; Hayashi, Y.; Nakashima, Y.; Kirikihira, T.;
Yamada, K.; Masuda, T. Effect of benzylic oxygen on the antioxidant
activity of phenolic lignans. J. Nat. Prod. 2005, 68, 1459−1470.
(15) Yamauchi, S.; Masuda, T.; Sugahara, T.; Kawaguchi, Y.; Ohuchi,
M.; Someya, T.; Akiyama, J.; Tominaga, S. Biosci. Biotechnol. Biochem.
2008, 72, 2981−2986.
AUTHOR INFORMATION
■
Corresponding Author
*E-mail: syamauch@agr.ehime-u.ac.jp. Fax: +81-89-977-4364.
Tel.: +81-89-946-9846.
(16) Kawaguchi, Y.; Yamauchi, S.; Masuda, K.; Nishiwaki, H.;
Akiyama, K.; Maruyama, M.; Sugahara, T.; Kishida, T.; Koba, Y.
5314
dx.doi.org/10.1021/jf5010572 | J. Agric. Food Chem. 2014, 62, 5305−5315

Journal of Agricultural and Food Chemistry
Article
Antimicrobial activity of stereoisomers of butane-type lignans. Biosci.
Biotechnol. Biochem. 2009, 73, 1806−1810.
(17) Nishiwaki, H.; Hasebe, A.; Kawaguchi, Y.; Akamatsu, M.; Shuto,
Y.; Yamauchi, S. Larvicidal activity of (−)-dihydroguaiaretic acid
derivatives against Culex pipiens. Biosci. Biotechnol. Biochem. 2011, 75,
1735−1739.
(18) Hasebe, A.; Nishiwaki, H.; Akiyama, K.; Sugahara, T.; Kishida,
T.; Yamauchi, S. Quantitative structure−activity relationship analysis
of antifungal (+)-dihydroguaiaretic acid using 7-phenyl derivatives. J.
Agric. Food Chem. 2013, 61, 8548−8555.
(19) Moss, G. P. Nomenclature of lignans and neolignans (IUPAC
recommendations 2000). Pure Appl. Chem. 2000, 72, 1493−1523.
5315
dx.doi.org/10.1021/jf5010572 | J. Agric. Food Chem. 2014, 62, 5305−5315