Myrsinoic acid E, an anti-inflammatory compound from Myrsine seguinii

Article abstract of DOI:10.1271/bbb.67.2038

The methanolic extract of Myrsine seguinii yielded the novel anti-inflammatory compound, myrsinoic acid E (1), whose structure was elucidated to be 3,5-digeranyl-4-hydroxy benzoic acid. We synthesized 1- and its 3,5-diprenyl (2) and 3,5-difarnesyl analogu

Full text of DOI:10.1271/bbb.67.2038

This article was downloaded by: [The University of Manchester Library]
On: 20 November 2014, At: 12:36
Publisher: Taylor & Francis
Informa Ltd Registered in England and Wales Registered Number: 1072954 Registered office: Mortimer
House, 37-41 Mortimer Street, London W1T 3JH, UK
Bioscience, Biotechnology, and Biochemistry
Publication details, including instructions for authors and subscription information:
http://www.tandfonline.com/loi/tbbb20
Myrsinoic Acid E, an Anti-inflammatory Compound
from Myrsine seguinii
Hidefumi MAKABEa, Shintaro MIYAZAKIb, Tsunashi KAMOb & Mitsuru HIROTAb
a Sciences of Functional Foods, Graduate School of Agriculture, Shinshu
UniversityMinami-minowa, Kami-ina, Nagano 399-4598, Japan
b Department of Bioscience and Biotechnology, Faculty of Agriculture, Shinshu
UniversityMinami-minowa, Kami-ina, Nagano 399-4598, Japan
Published online: 22 May 2014.
To cite this article: Hidefumi MAKABE, Shintaro MIYAZAKI, Tsunashi KAMO & Mitsuru HIROTA (2003) Myrsinoic Acid E, an
Anti-inflammatory Compound from Myrsine seguinii , Bioscience, Biotechnology, and Biochemistry, 67:9, 2038-2041, DOI:
10.1271/bbb.67.2038
To link to this article: http://dx.doi.org/10.1271/bbb.67.2038
PLEASE SCROLL DOWN FOR ARTICLE
Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) contained
in the publications on our platform. However, Taylor & Francis, our agents, and our licensors make no
representations or warranties whatsoever as to the accuracy, completeness, or suitability for any purpose of
the Content. Any opinions and views expressed in this publication are the opinions and views of the authors,
and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be relied
upon and should be independently verified with primary sources of information. Taylor and Francis shall
not be liable for any losses, actions, claims, proceedings, demands, costs, expenses, damages, and other
liabilities whatsoever or howsoever caused arising directly or indirectly in connection with, in relation to or
arising out of the use of the Content.
This article may be used for research, teaching, and private study purposes. Any substantial or systematic
reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any
form to anyone is expressly forbidden. Terms & Conditions of access and use can be found at http://
www.tandfonline.com/page/terms-and-conditions

Biosci. Biotechnol. Biochem., 67 (9), 2038–2041, 2003
Note
Myrsinoic Acid E, an Anti-in‰ammatory Compound from Myrsine seguinii
Hidefumi MAKABE,
† Shintaro MIYAZAKI,2 Tsunashi KAMO,2 and Mitsuru HIROTA
1,
2
1Sciences of Functional Foods, Graduate School of Agriculture, Shinshu University, Minami-minowa,
Kami-ina, Nagano 399-4598, Japan
2Department of Bioscience and Biotechnology, Faculty of Agriculture, Shinshu University, Minami-minowa,
Kami-ina, Nagano 399-4598, Japan
Received April 17, 2003; Accepted May 22, 2003
The methanolic extract of Myrsine seguinii yielded
the novel anti-in‰ammatory compound, myrsinoic acid
E (1), whose structure was elucidated to be 3,5-diger-
anyl-4-hydroxy benzoic acid. We synthesized 1- and its
3,5-diprenyl (2) and 3,5-difarnesyl analogues (3). Com-
myrsinoic acid A (4).3) In the 1H-NMR spectrum, the
only diŠerences were the presence in 1 of an oleˆnic
＝
6.7 Hz,
methine triplet (H-6
2H) and a symmetrical three-methyl singlet at 1.76,
1.68, and 1.60 (H-9 , -9 , -8 , -8 , -10 , -10 ). These
?, -6!
) at
d
5.08 (t,
J
?
!
?
!
?
!
pounds 1–3 suppressed 12-
acetate (TPA)-induced in‰ammation of mouse ears by
59 , 14 , and 69 at a dose of 1.4 mol.
O
-tetradecanoylphorbol-13-
data indicate that compound 1 possessed two sym-
metric geranyl group at the 3, 5 position of 4-hydroxy
benzoic acid. The 13C-NMR spectrum of 1 was in
complete accordance with the proposed structure.
The synthesis of compound 1 and its analogues 2
and 3 is shown in Scheme 1. 4-Iodophenol (5) was
used as a starting material. The introduction of a 3,5-
digeranyl group was performed by using Hori's
procedure to give 6.5) Pd-catalyzed carbonylation6)
%
%
%
m
Key words: anti-in‰ammatory; Myrsine seguinii
;
12 - - tetradecanoylphorbol - 13 - acetate
O
(TPA)-induced edema; Pd-catalyzed
carbonylation
Skin tumor promoters such as 12-
O
-tetradecanoyl-
aŠorded ester 9 in a 91
z
yield. Finally, hydrolysis of
yield. The spectral
phorbol-13-acetate (TPA) induce edema on the skin
with cell proliferation.1,2) Inhibitors of TPA-induced
edema can be expected to suppress various physiolog-
ical responses involving cell proliferation as well as
in‰ammation. In a search for new types of anti-
in‰ammatory compounds, we found that the metha-
nolic extract of Myrsine seguinii showed potent anti-
in‰ammatory activity. We have already reported the
isolation and identiˆcation of four novel related
compounds, myrsinoic acid A (4),3) B, C and F.4)
In our continuing search for other anti-in‰amma-
tory compounds from this plant, we isolated the
novel terpeno-benzoic acid, myrsinoic acid E (1).
However, we could not evaluate its anti-in‰ammato-
ry activity because of its small amount. This prompt-
ed us to synthesize compound 1, as well as several
analogues for further anti-in‰ammatory activity
studies.
the methyl ester gave 1 in an 88
z
data for synthetic 1 are in good accordance with
those of natural 1. To examine the importance of the
C-3 and C-5 side chain length on the anti-in‰amma-
tory activity, we designed target molecule 2 with a
prenyl group and 3 with a farnesyl group. The syn-
thesis of 2 and 3 was performed as just described.
The anti-in‰ammatory activities of compounds
1–4, and 4-hydroxybenzoic acid are summarized in
Table 1. Compounds 1, 3 and 4 showed strong anti-
in‰ammatory activity, while compound 2 and 4-
hydroxybenzoic acid exhibited low activity. The
long terpene moieties of terpeno-benzoic acid-type
compounds seem to have been essential for the anti-
in‰ammatory activity.
Experimental
The methanolic extract of M. seguinii yielded
myrsinoic acid E (1). The HR-EIMS analysis of this
compound gave the molecular formula of C27H38O3
(found 410.2836) with nine unsaturations. The IR
spectrum of 1 showed the presence of hydroxyl
Instruments. The following instruments were used:
1
a Bruker DR 500 FT-NMR spectrometer for H- and
13C-NMR spectra, a Jeol JMS 700 mass spectrometer
for mass spectra, and a Jasco FT IR-480 Plus for IR
W
spectra.
(3435 cm－1) and carboxyl (1683 cm－ and 3200–
1
2400 cm－1) groups. The 1H-NMR and 13C-NMR
Anti-in‰ammatory test. Mouse in‰ammatory tests
spectra of 1 showed close similarity with those of
were conducted by Gshwendt's method.7) TPA was
†
To whom correspondence should be addressed. Fax:
＋
81-265-77-1700; E-mail: makabeh
＠
gipmc.shinshu-u.ac.jp
Abbreviations: DMF,
N
,
N
-dimethylformamide; TPA, 12-O
-tetradecanoylphorbol-13-acetate

Myrsinoic Acid E from Myrsine seguinii
2039
Table 1. Anti-in‰ammatory Activities of Compounds 1–4, and
Benzoic Acid Derivatives in the Mouse Ear In‰ammatory Test
Inhibitory eŠect
Test compound
(IE
z)
±
1
2
3
4
59
14
69
65
2
z(
z(
z(
z(
z(
0.79)
1.15)
1.17)
1.03)
0.38)
±
±
±
±
*
p
-hydroxybenzoic acid
In parentheses: A sample (1.4
after 30 min, TPA (0.5 g) was applied to both ears of the mouse. The ede-
ma was evaluated after 7 hr, the inhibitory eŠect being expressed as the per-
mmol) was applied on one mouse ear and,
m
*
centage ratio of the edema. Five mice were used for each experiment. At a
dose of 500 g (3.6 mol).
m
m
(M＋ ): calcd: for C27 H38O3, 410.2821; found,
1
410.2836. H-NMR (500 MHz, CDCl3)
2H, H-2, -6), 5.36 (s, 1H, -OH), 5.32 (t,
d: 7.74 (s,
＝
J
6.6 Hz,
, -6 ),
), 2.09 (m, 8H,
), 1.76 (s, 6H, H-9 , -9 ), 1.68 (s,
), 1.60 (s, 6H, H-10 , -10
). 13C-NMR
(125 MHz, CDCl3) : 170.44 (C-7), 158.01 (C-4),
138.84 (C-3 , -3 ), 131.94 (C-7 , -7 ), 130.46 (C-2,
-6), 127.25 (C-3, -5), 123.86 (C-6 , -6 ), 121.26
(C-2 , -2 ), 120.92 (C-1), 39.73 (C-4 , -4 ), 29.48
(C-1 , -1 ), 26.46 (C-5 , -5 ), 25.68 (C-8 , -8 ),
17.72 (C-10 , -10 ), 16.24 (C-9
＝
2H, H-2
3.38 (d,
H-4 , -4
6H, H-8
?
, -2
6.8 Hz, 4H, H-1
, -5 , -5
, -8
!
), 5.08 (t,
J
6.7 Hz, 2H, H-6
?
!
J
＝
?
, -1
!
?
!
?
!
!
?
!
?
?
!
Fig. 1. Structures of Myrsinoic Acid E and Its Analogues.
d
?
!
?
!
?
!
?
!
?
!
?
!
?
!
?
!
?
!
?, -9!
). IR nmax (ˆlm)
cm－ : 3435 (OH), 3200–2400 (COOH), 1683 (C O),
1
＝
1601 (C
＝
C), 1436, 1260, 800.
Dialkylation of 4-Iodophenols. 3,5-Dialkyl-4-
iodophenols were prepared according to the reported
procedure.5)
Scheme 1. Reagents and Conditions.
(a) 1.0 eq. of alkyl bromide, 10
z
NaOH; (b) 5 molz
Cl2Pd(PPh3)2, CO (1 atm), 2.0 eq. of Et3N, 4.0 eq. of MeOH;
(c) NaOH, MeOH, re‰ux.
(2
?
E,2
!
E)-4-Iodo-2-(3
?
,7
?
-dimethylocta-2
?
,6
?
-
dienyl)-6-(3
!
,7 -dimethylocta-2
!
!
,6!
-dienyl)phenol
(
6
). IR (ˆlm)
n
max cm－1: 3456, 2968, 2924, 1668, 1458,
1
1198. H-NMR (500 MHz, CDCl3)
d
: 7.26 (s, 2H),
purchased from Sigma Chemical Co. This experi-
ment complied with the regulations concerning
animal experimentation and the care of experimental
animals of the Faculty of Agriculture at Shinshu
University.
5.36 (s, 1H, -OH), 5.27 (t,
J
＝
7.1 Hz, 2H), 5.08 (t,
J
＝
＝
7.1 Hz, 4H), 2.00–2.20
6.6 Hz, 2H), 3.29 (d,
J
(m, 8H), 1.74 (s, 6H), 1.69 (s, 6H), 1.60 (s, 6H). 13C-
NMR (125 MHz, CDCl3)
d: 152.84, 138.64, 136.28,
131.94, 129.97, 123.85, 121.21, 82.69, 39.70, 29.15,
26.42, 25.74, 17.73, 16.19. HREIMS m z (M＋ ):
W
Isolation of compound
1
.
Fresh leaves and twigs of
calcd. for C26H37OI, 492.1889; found, 492.1901.
M. seguinii (12 kg) were extracted with methanol,
and the extract was concentrated in vacuo. The aque-
ous solution was successively partitioned with hexane
water and ethyl acetate water. Strong activity was
found in the hexane-soluble portion (69 g), which
was chromatographed in a silica gel column with
4-Iodo-2-(3
butenyl)phenol (
with those reported.5)
?
-methyl-2
?-butenyl)-6-(3!-methyl-2!-
7
). 1H-NMR data were identiˆed
W
W
(2
trimethylocta-2
trimethylocta-2 ,6
max cm－1: 3459, 2966, 2922, 2853, 1665, 1457, 1198.
1H-NMR (500 MHz, CDCl3)
: 7.26 (s, 2H), 5.34 (s,
7.1 Hz, 2H), 5.08 (m, 4H), 3.29 (d,
7.1 Hz, 4H), 1.95–2.20 (m, 16H), 1.74 (s, 6H),
?
E, 6
?
E,2
!
E,6
,6
,10
!
E)-4-Iodo-2-(2
,10 -trienyl)-6-(2
-trienyl)phenol (
?
!
,6
?
!
,10
?
!
-
hexane ethyl acetate by 10
z
stepwise elution. The
?
!
?
?
,6
,10
-
W
20
matographed on ODS with gradient elution from
70 methanol water to methanol. The active frac-
tion was further puriˆed by HPLC (95
water) to give compound 1 (1.6 mg). HREIMS m z
z
ethyl acetate hexane eluate (13.8 g) was chro-
!
!
8). IR (ˆlm)
W
n
z
d
W
＝
z
methanol
1H), 5.27 (t,
＝
J
J
W
W

2040
H. MAKABE et al.
1.67 (s, 6H), 1.60 (s, 12H). 13C-NMR (125 MHz,
CDCl3) : 152.82, 138.69, 136.28, 135.53, 131.27,
Hydrolysis of methyl ester. 4-Hydroxy-3,5-
dialkylated benzoic acids were prepared according to
the reported procedure apart from using NaOH.3)
d
129.92, 125.91, 124.40, 123.74, 121.18, 82.70, 29.15,
26.76, 26.44, 26.38, 25.69, 17.71, 16.25, 16.06.
HREIMS m z (M＋ ): calcd. for C H OI, 628.3141;
(2
dienyl)-5-(3
acid (
). IR, 1H-NMR, 13C-NMR and HREIMS were
identiˆed with those of myrsinoic acid E (1).
?
E,2
!
E)-4-hydroxy-3-(3
?
,7
?
-dimethylocta-2
?,6?-
W
36
53
found, 628.3131.
!
,7 -dimethylocta-2
!
! !
,6 -dienyl)benzoic
1
Carbonylation of 2,6-dialkylated 4-iodophenols.
To solution of 2,6-dialkylated 4-iodophenol
(0.5 mmol) in 1 ml of -dimethylformamide
(DMF) were successively added NEt3 (0.14 ml,
1 mmol), MeOH (64 mg, 2.0 mmol), and
Cl2Pd(PPh3)2 (0.025 mmol, 17.6 mg). The mixture
was charged with CO (1 atm), heated to 60 C, and
stirred for 6 h. The usual work up gave a pure methyl
ester after column chromatography (hexane ethyl
a
N
,
N
4-Hydroxy-3-(3
?
-methyl-2
?
-butenyl)-5-(3
!
-methyl-
1
2
!
-butenyl)benzoic acid (
2
). IR (ˆlm) nmax cm－
:
3437, 3200–2800, 2960, 2926, 2856, 1687, 1603, 1434,
1
1287, 713. H-NMR (500 MHz, CDCl3)
d
: 7.76 (s,
7.2 Hz, 2H), 3.37 (d,
＝
7.2 Hz, 4H), 1.78 (s, 6H), 1.79 (s, 6H). 13C-NMR
＝
2H), 5.90 (br., 1H), 5.32 (t, J
9
J
(125 MHz, CDCl3)
d: 171.57, 157.86, 135.21,
W
＝
acetate 8:1).
130.47, 127.16, 121.30, 121.08, 51.77, 29.55, 25.82,
17.92. HREIMS m z (M＋ ): calcd. for C17H22O3,
W
(2
ta-2
dienyl)benzoate (
?
?
E,2
!
?
E)-Methyl4-hydroxy-3-(3
-dienyl)-5-(3 ,7 -dimethylocta-2
). IR (ˆlm) nmax cm－1: 3437, 2966,
?
,7
?
-dimethyloc-
274.1569; found, 274.1597.
,6
!
!
!
,6
!-
9
(2
?
E,6
?
E,2
!
E,6
,6
,10
!
?
E)-4-Hydroxy-3-(3
?
,7
?
,11
?
!
-
-
2919, 2855, 1718, 1696, 1603, 1434, 1317, 1280, 1199,
trimethylocta-2
?
,6!
,10 -trienyl)-5-(3
?
!
,7
!
,11
1
771. H-NMR (500 MHz, CDCl3)
d
: 7.70 (s, 2H),
trimethylocta-2
!
!-trienyl)benzoic acid (3). IR
5.84 (s, 1H), 5.31 (t,
J
7.0 Hz, 2H), 5.06 (t,
J
(ˆlm) nmax cm－1: 3434, 3200–2800, 2965, 2922, 2856,
＝
＝
1
6.6 Hz, 2H), 3.88 (s, 3H), 3.39 (d,
2.05–2.15 (m, 8H), 1.76 (s, 6H), 1.68 (s, 6H), 1.60 (s,
6H). 13C-NMR (125 MHz, CDCl3)
: 167.27, 157.25,
J
＝
7.0 Hz, 4H),
1683, 1602, 1438, 1412, 1281, 1207, 775. H-NMR
(500 MHz, CDCl3)
5.32 (t,
d
: 7.77 (s, 2H), 5.91 (br., 1H),
＝
＝
d
J
7.0 Hz, 2H), 5.10 (m, 4H), 3.37 (d, J
138.65, 131.90, 129.76, 127.13, 123.89, 122.01,
121.42, 51.76, 39.74, 29.44, 26.48, 25.69, 17.71,
7.0 Hz, 4H), 1.95–2.20 (m, 16H), 1.78 (s, 6H), 1.66
(s, 6H), 1.60 (s, 6H), 1.59 (s, 6H). 13C-NMR (125
16.24. HREIMS m z (M＋ ): calcd. for C28H40O3,
MHz, CDCl3) d: 171.43, 157.99, 138.88, 135.53,
W
424.2977; found, 424.2971.
131.26, 130.47, 127.22, 124.41, 123.76, 121.23,
121.00, 39.75, 39.71, 29.47, 26.76, 26.47, 25.68,
Methyl 4-hydroxy-3-(3
?
-methyl-2
?
-butenyl)-5-(3
!
:
-
17.68, 16.30, 16.05. HREIMS m z (M＋ ): calcd. for
W
1
methyl-2 -butenyl)benzoate (10). IR (ˆlm)
!
n
max cm－
C37H54O3, 546.4073; found, 546.4084.
3439, 2966, 2921, 2853, 1718, 1697, 1602, 1434, 1317,
1279, 1197, 771. 1H-NMR (500 MHz, CDCl3)
d
: 7.69
7.2 Hz, 2H), 3.36 (d,
7.2 Hz, 4H), 1.95–2.20 (m, 16H), 1.78 (s, 12H).
Acknowledgments
＝
J
(s, 2H), 5.81 (s, 1H), 5.31 (t,
J
＝
This work was supported in part by grant aid from
Japan Society for the Promotion of Science
(13760085 to H. M.). We also thank Ms. Keiko
Hashimoto of the Faculty of Agriculture at Shinshu
University for the 500 MHz NMR measurements.
13C-NMR (125 MHz, CDCl3)
d: 167.26, 157.09,
135.02, 129.77, 127.04, 122.08, 121.45, 51.77, 29.61,
25.82, 17.92. HREIMS m z (M＋ ): calcd. for
W
C18H24O3, 288.1725; found, 288.1736.
Methyl
,7 ,11
,7 ,11
(2
?
E,6
?
E,2
!
E,6
,6
,10
!
E)-4-hydroxy-3-
,10 -trienyl)-5-
References
(3
(3
?
!
?
!
?
!
-trimethylocta-2
?
!
?
?
1) Hecker, E., Structure-activity relationships in diter-
pene esters irritant and cocarcinogenic to mouse skin.
In Carcinogenesis, vol. 2, ``Mechanisms of Tumor
Promotion and Cocarcinogenesis'', eds. Slaga, T. J.,
Sivak, A., and Boutwell, R. K., Raven Press, New
York, pp. 11–48 (1978).
2) Fischere, S. M., Cameron, G. S., Baldwin, J. K.,
Jasheway, D. W., Patrick, K. E., and Belury, M. A.,
The arachidonic acid cascade and multistage carcino-
genesis in mouse skin. In ``Skin Carcinogenesis:
Mechanisms and Human Relevance'', eds. Slaga, T.
J., Klein-Szanto, A. J. P., Boutwell, R. K., Stevenson,
D. E., Spitzer, H. L., and D'Motto, B., Alan R. Liss,
New York, pp. 249–264 (1989).
-trimethylocta-2
!
,6
!-trienyl)benzoate
(
11). IR (ˆlm) nmax cm－1: 3437, 2966, 2919, 2854,
1
1719, 1697, 1603, 1435, 1317, 1281, 1198, 771. H-
NMR (500 MHz, CDCl3)
d: 7.72 (s, 2H), 5.80 (s,
＝
＝
1H), 5.32 (t,
3H), 3.37 (d,
J
6.9 Hz, 2H), 5.09 (m, 4H), 3.88 (s,
6.9 Hz, 4H), 1.95–2.20 (m, 16H),
J
1.77 (s, 6H), 1.66 (s, 6H), 1.60 (s, 6H), 1.59 (s, 6H).
13C-NMR (125 MHz, CDCl3)
d
: 167.24, 157.20,
138.70, 135.49, 131.26, 129.76, 127.09, 124.41,
123.79, 122.05, 121.37, 51.73, 39.76, 39.71, 29.62,
29.50, 26.76, 26.50, 25.68, 17.68, 16.30, 16.04.
HREIMS m z (M＋ ): calcd. for C H O , 560.4229;
W
38
56
3
found, 560.4239.

Myrsinoic Acid E from Myrsine seguinii
2041
3) Dong, M., Nagaoka, M., Miyazaki, S., Iriye, R., and
Hirota, M., 3-Geranyl-4-hydroxy-5-(3 -methyl-2
butenyl)benzoic acid as an anti-in‰ammatory com-
pound from Myrsine seguinii Biosci. Biotechnol.
Biochem., 63, 1650–1653 (1999).
4) Hirota, M., Miyazaki, S., Minakuchi, T., Takagi, T.,
and Shibata, H., Myrsinoic acids B, C and F, anti-in-
artepillin C established by o,o?-diprenylation of p-
halophenols in water. J. Org. Chem., 67, 2355–2357
(2002).
?
?-
.
6) Schoenberg, A., and Heck, R. F., Palladium-catalyzed
amidation of aryl, heterocyclic, and vinylic halides. J.
Org. Chem., 43, 3327–3331 (1974).
7) Gschwendt, M., Kittstein, W., Furstenberger, G., and
Marks, F., The mouse ear edema: A quantitatively
evaluable assay for tumor promoting compounds and
for inhibitors of tumor promotion. Cancer Lett., 25,
177–185 (1984).
‰ammatory compounds from Myrsine seguinii
Biotechnol. Biochem., 66, 655–659 (2002).
. Biosci.
5) Uto, Y., Hirata, A., Fujita, T., Takubo, S.,
Nagasawa, H., and Hori, H., First total synthesis of