Antiallergic activity of rosmarinic acid esters is modulated by hydrophobicity, and bulkiness of alkyl side chain

Article abstract of DOI:10.1080/09168451.2015.1010478

Methyl, propyl and hexyl esters of rosmarinic, caffeic and p-coumaric acids were tested for antiallergic activity, and rosmarinic acid propyl ester exhibited the greatest β-hexosaminidase release suppression (IC₅₀, 23.7 μM). Quadratic correlations between pIC₅₀ and cLogP (r² = 0.94, 0.98, and 1.00, respectively) were observed in each acid ester series. The antiallergic activity is modulated by hydrophobicity, and alkyl chain bulkiness.

Full text of DOI:10.1080/09168451.2015.1010478

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Antiallergic activity of rosmarinic acid esters is
modulated by hydrophobicity, and bulkiness of alkyl
side chain
Fengxian Zhua, Zhongming Xub, Lina Yonekuraac, Ronghua Yangb & Hirotoshi Tamuraac
a The United Graduate School of Agricultural Sciences, Ehime University, Matsuyama,
Japan
b College of Food Science and Biotechnology, Zhejiang Gongshang University, Hangzhou,
China
c The Graduate School of Agriculture, Kagawa University, Miki-cho, Kagawa, Japan
Published online: 16 Feb 2015.
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To cite this article: Fengxian Zhu, Zhongming Xu, Lina Yonekura, Ronghua Yang & Hirotoshi Tamura (2015): Antiallergic
activity of rosmarinic acid esters is modulated by hydrophobicity, and bulkiness of alkyl side chain, Bioscience,
Biotechnology, and Biochemistry, DOI: 10.1080/09168451.2015.1010478
To link to this article: http://dx.doi.org/10.1080/09168451.2015.1010478
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Bioscience, Biotechnology, and Biochemistry, 2015
Note
Antiallergic activity of rosmarinic acid esters is modulated by hydrophobicity,
and bulkiness of alkyl side chain
Fengxian Zhu1, Zhongming Xu2, Lina Yonekura1,3, Ronghua Yang2 and Hirotoshi Tamura1,3,
*
2
1The United Graduate School of Agricultural Sciences, Ehime University, Matsuyama, Japan; College of Food
3
Science and Biotechnology, Zhejiang Gongshang University, Hangzhou, China; The Graduate School of
Agriculture, Kagawa University, Miki-cho, Kagawa, Japan
Received November 25, 2014; accepted January 13, 2015
http://dx.doi.org/10.1080/09168451.2015.1010478
Methyl, propyl and hexyl esters of rosmarinic,
caffeic and p-coumaric acids were tested for
antiallergic activity, and rosmarinic acid propyl
ester exhibited the greatest β-hexosaminidase release
suppression (IC50, 23.7 μM). Quadratic correlations
between pIC50 and cLogP (r2 = 0.94, 0.98, and 1.00,
respectively) were observed in each acid ester
series. The antiallergic activity is modulated by
hydrophobicity, and alkyl chain bulkiness.
screening for anti-allergic reagents.10,11) Regarding
inhibition of degranulation of RBL 2H3 cells by phe-
nolic compounds, so far there are four possible mecha-
nisms. The first possibility is the inhibition of binding
between the FcεRI receptor and an allergen–IgE com-
plex. Apple extract and brown algae phlorotannins inhi-
bit binding between IgE and FcεRI receptor,
suppressing mast cell activation.12,13) The second is
due to binding inhibition between a cross-linked IgE/
FcεRI receptor and the antigen.14) The third is to form
insoluble complexes or aggregates with the allergens,
inhibit their binding to IgE, and reducing their aller-
genic capacity.15) The fourth one is the suppression of
FcεRI receptor expression in mast cells.16)
Key words: rosmarinic acid; esterification; antiallergic
activity; hydrophobicity; bulkiness
We recently isolated a RA-rich extract with high
anti-allergic activity from Perilla leaves using a supra-
molecular separation method.17) In that study, isolation
of RA was attained without the use of preparative
HPLC, therefore, enabling mass production of RA at a
relatively lower cost and widening the possibilities for
its use as a functional ingredient in food industries. RA
methyl ester, a minor component in Perilla leaf extract,
suppressed β-hexosaminidase release 14.3 times greater
than RA.12) Therefore, the esterification of such phenyl-
propanoids is highly expected to enhance antiallergic
activity. In this paper, we explored the potential of RA
and related compounds as therapeutic agents for type I
allergies, and investigated the effects of esterification as
an approach to enhance their antiallergic activity. As
the assay we have used begins by sensitizing the mast
cells with IgE, suppression of degranulation can be
attributed to the inhibition of binding of a cross-linked
IgE/FcεRI receptor and the antigen.
Rosmarinic acid (RA), a major bioactive compound
in rosemary, mint, basil, and perilla exerts a variety of
biological activities, such as antioxidant1) and anti-
inflammatory2) activities, and reducing liver injury.3)
Additionally, RA has been shown to suppress inflam-
mation and to alleviate the symptoms of type I allergic
conditions, such as allergic asthma4) and seasonal aller-
gic rhinoconjunctivitis, in animals and humans.5,6) RA
suppresses allergic inflammation by inhibiting IgE
response, inflammatory cytokine production, and COX-
2 expression.2)
Mast cells, widely distributed throughout the body as
a part of the immune system, play an important role in
both immediate and late-phase response of type I
allergy. The activation of mast cells is initiated by anti-
gen-induced cross-linking of IgE to IgE receptors
(FcεRI),7,8) followed by a series of signaling events,
leading to phosphorylation of tyrosine kinases, and
mobilization of Ca2+ into cytosol.9) Within minutes of
activation, mast cells secret several mediators, such as
β-hexosaminidase and histamine, that result in acute
inflammation and chronic allergic disease.7) The RBL-
2H3 mast cell line is considered an excellent model
system for the study of mast cell degranulation10) and
Esters of RA, caffeic acid (CA), p-coumaric acid
(Cou), ferulic acid (FA), and cinnamic acid (Cin) were
prepared by reaction with methanolic HCl. Three to
four hundred milligrams of the free acids were individ-
ually dissolved in 25 mL methanol containing 1% HCl.
*Corresponding author. Email: tamura@ag.kagawa-u.ac.jp
Abbreviations: RA, rosmarinic acid; RA-me, rosmarinic acid methyl ester; RA-pro, rosmarinic acid propyl ester; RA-hex, rosmarinic acid hexyl
ester; CA, caffeic acid; CA-me, caffeic acid methyl ester; CA-pro, caffeic acid propyl ester; CA-hex, caffeic acid hexyl ester; Cou, p-coumaric acid;
Cou-me, p-coumaric acid methyl ester; Cou-pro, p-coumaric acid propyl ester; Cou-hex, p-coumaric acid hexyl ester, FA,ferulic acid; FA-me, feru-
lic acid methyl ester; Cin, cinnamic acid; Cin-me, cinnamic acid methyl ester; cLogP, calculated logarithmic value of octanol–water partition coef-
ficient; pIC50, −logIC50
.
© 2015 Japan Society for Bioscience, Biotechnology, and Agrochemistry

2
F. Zhu et al.
The reaction was carried out at room temperature and
monitored by ultrafast liquid chromatography. The reac-
tion solution was diluted with water (50 mL) and
extracted with diethyl ether (50 mL) in a separating
funnel. After the diethyl ether layer was collected and
washed with water and 5% NaHCO3 aqueous solution,
the organic layer was evaporated to dryness. The pro-
pyl and hexyl esters were synthesized as described
above, using 1-propanol and 1-hexanol in place of
methanol.
RA, CA, Cou, FA, Cin, and their methyl esters sup-
pressed β-hexosaminidase release in a dose-dependent
manner up to the concentration of 2.0 mM, as shown
in Fig. 1. The IC50 values for all methyl esters were
below 0.4 mM (RA-me, 0.06 mM; CA-me, 0.12 mM;
Cou-me, 0.18 mM; FA-me, 0.14 mM; and Cin-me,
0.33 mM), and thus, smaller than the IC50 values of all
free acids (RA, 1.31 mM; CA, 1.46 mM; Cou,
1.04 mM; FA, 0.98 mM; and Cin, 1.50 mM). RA-me
exerted the greatest β-hexosaminidase release suppres-
sion (IC50, 0.06 mM), followed by CA-me (IC50,
0.12 mM) among all of the methyl esters. CA-me sup-
pressed β-hexosaminidase release similar to RA-me,
and then similar suppression activity was observed
between RA (1.31 mM) and CA (1.46 mM). It is indi-
cated that β-hexosaminidase release suppression of RA
and its methyl ester is mainly due to its CA moiety,
which may interact with a cross-linked IgE/FcεRI
receptor of RBL-2H3 cells. This is partly supported by
the results that 3-(3,4-dihydroxyphenyl)lactic acid (a
similar moiety of RA partial structure) related com-
pounds did not suppress β-hexosaminidase release at
all.
As a greater suppression of β-hexosaminidase release
was achieved by methyl esterification of such phenyl-
propanoids was observed, we investigated the effects of
longer alkyl chain esters of RA and its related phenyl-
propanoids on β-hexosaminidase release suppression.
Furthermore, RA-me, RA-pro, and RA-hex suppressed
β-hexosaminidase release to a greater extent than the
corresponding esters of CA and Cou (Table 1). Among
all free acids and esters, rosmarinic acid propyl ester
(RA-pro, IC50, 23.7 μM) exhibited the greatest
suppression on β-hexosaminidase release, followed by
rosmarinic acid hexyl ester (RA-hex, IC50, 34.5 μM),
caffeic acid propyl ester (CA-pro, IC50, 55.3 μM), and
rosmarinic acid methyl ester (RA-me, IC50, 57.3 μM),
as shown in Table 1. The activity of rosmarinic acid
propyl ester was only 9.1 times lower than luteolin
(IC50, 2.6 μM), a flavonoid with high antiallergic
activity that was tested in a parallel experiment of our
laboratory,19) but increased 55.4 times compared with
the activity of RA.
In addition, it was recently found that RA can be
easily extracted from natural resources in a large scale
with a newly established isolation method from Perilla
leaves using a supramolecular technique,17) implying
that RA, the precursor of RA esters, can be easily sup-
plied from natural resources such as Perilla leaves as
well as other Lamiaceae family plants that are rich in
RA. Furthermore, for applications in the food indus-
tries, we suggest that the ethyl esters might be a good
choice as their synthesis can be carried out with etha-
nol, regarded as a safe ingredient in food manufactur-
ing. Alcoholic beverages or other processed foods
containing Perilla leaves may bring great benefit for
alleviating allergy symptoms.
The octanol–water partition coefficient (LogP), a
measurement of molecular hydrophobicity, is a vital
parameter for membrane permeability, bioavailability,
and hydrophobic drug–receptor interactions. Regarding
the interaction between polyphenols and receptors, iso-
flavones are known to bind estrogen receptors and the
hydrophobic aglycone binds more than their
The antiallergic activity of the free acids and synthe-
sized esters was assessed by their ability to inhibit β-
hexosaminidase release from RBL-2H3 cells, using the
following method.17,18) In brief, the RBL-2H3 cells
were precultured in a 24-well plate (2.5 × 105 cells/
well) at 37 °C under a humidified 5% CO2 atmosphere
overnight. The cells were washed with PBS and incu-
bated with 500 μL antibody solution (mouse monoclo-
nal anti-dinitrophenyl antibody, 50 ng/mL) for 2 h’
sensitization. A test sample solution of 490 μL was
added to each well after washing with modified
Tyrode’s buffer (MT buffer). The MT buffer was used
as control instead of the sample. Albumin dinitrophenyl
(10 μL, final concentration = 50 ng/mL) was added to
each well to evoke allergic reactions (degranulation) of
the cells for 30 min. The supernatants (50 μL) were
transferred into wells (96-well microplate) and incu-
bated with 100 μL substrate (3.3 mM p-nitrophenyl-2-
acetamide-2-deoxy-β-D-glucopyranoside) at 37 °C for
25 min. The absorbance (OD) was measured at 405 nm
using a microplate reader. The gaining OD reflects β-
hexosaminidase release. The calculation was performed
using Equations (1) and (2) below. In “blank,” neither
antibody solution nor sample was added to cells to
account for spontaneous β-hexosaminidase release from
cells. In “control,” MT buffer instead of samples was
added to cells to account for β-hexosaminidase release
from cells in conditions without a sample. In “total,”
cells were lysed by 0.1% Triton X-100 in MT buffer to
measure the total amount of β-hexosaminidase con-
tained in the cells. In “sample,” both antibody solution
and samples were added to cells to measure β-hexosa-
minidase release from cells in the presence of our test
compounds.
ratio of b ꢀ hexosaminidase release ð%Þ
ðODcontrol or ODsample ꢀ ODblank
Þ
¼
ꢁ 100
(1)
ðODtotal ꢀ ODblank
Þ
The ratio of β-hexosaminidase release (%) of “con-
trol” against “total” (Triton X-100) ranged from 29 to
50%. Relative rate of β-hexosaminidase release of
“sample” against “control” was expressed as β-hexosa-
minidase release (%) as shown below,
b-hexosaminidase release ð%Þ
ODsample ꢀ ODblank
¼
ꢁ 100
(2)
ODcontrol ꢀ ODblank
The data are presented as mean standard derivation
(SD) of quadruplicate wells. Statistical differences were
assessed using one-way ANOVA, followed by the
New–Keuls test. p values below 0.05 were considered
statistically significant. Statistical analysis was carried
out by GraphPad PRISM 5.

Rosmarinic acid esters contribute to antiallergic activity
3
Fig. 1. Antiallergic activity of RA, its related compounds, and their methyl esters by β-hexosaminidase release suppression.
Note: Each IC50 value represents mean SD in quadruplicate. Abbreviation of compounds shown here was followed the abbreviation list.
Table 1. β-Hexosaminidase release suppression and calculated
octanol–water partition coefficient of esters of RA, CA, and p-coumaric
acid.
the acid esters series, with correlation coefficients equal
to 0.94 (p < 0.05) for the RA series, 0.98 (p < 0.01)
for the CA series, and 1.00 (p < 0.01) for the Cou ester
series (Fig. 2). LogP values for maximal suppression of
β-hexosaminidase release were calculated from the indi-
vidual quadratic equations (Fig. 2) as 3.7 for RA esters,
2.6 for CA esters, and 2.9 for Cou esters. The alkyl
esters chain lengths deduced from these values would
correspond to butyl ester for RA, propyl ester for CA,
and propyl ester for Cou (Fig. 2). Furthermore, RA
butyl ester is expected to have a higher pIC50 value
than that was observed for the propyl ester, as deduced
from the quadratic equation (y = −0.47 x2 + 3.44 x − 1.
35) to be 11.4 μM. The quadratic equations reflect the
lowering of antiallergic effects of hexyl esters after the
very strong effects of shorter esters. Our results showed
that the hydrophobic propyl esters of RA, CA, and
Cou strongly suppressed β-hexosaminidase release,
thus, it is possible that propyl esters have higher affin-
ity to the cross-linked IgE/FcεRI receptor, while the
bulky hexyl esters would bind less efficiently. It has
already been reported that steric effects of ligands can
interfere with the binding to the cross-linked IgE/FcεRI
receptor, influencing mast cell degranulation.22,23)
Regarding enzyme reactions, a quadratic correlation
between the hydrophobicity of 4-sulfamate derivatives
with different alkyl chain length and estrone sulfa-
tase inhibition has been reported.24) Furthermore,
Handlogten et al. pointed out that heterobivalent
ligands selectively bind to IgE, and competitively inhi-
bit allergen binding,14) similar to enzyme reaction.
Taken together, those reports support the quadratic
correlation between hydrophobicity and antiallergic
effect found in our study, and provide some evidence
that hydrophobicity and steric effects play a role in the
β-hexosaminidase suppressive effect within each of our
phenolic acid esters series.
A
Compounds
IC50 (μM)
cLogP
RA
1313.7 189.2d
57.3 4.0a
1.6
2.2
3.1
4.7
0.9
1.6
2.4
4.0
1.4
2.1
2.9
4.5
RA-me
RA-pro
RA-hex
CA
CA-me
CA-pro
CA-hex
Cou
23.7 2.5a
34.5 1.8a
1457.0 35.1d
120.4 0.1a
55.3 3.3a
523.1 14.7b
1039.8 252.4c
182.4 6.8a
78.6 8.2a
Cou-me
Cou-pro
Cou-hex
1162.7 99.0c
AIC50 values were calculated from dose-dependent curves of β-hexosamini-
dase suppression using Graphpad prism software. Each IC50 value represents
mean SD in quadruplicate. Means with different superscript lower case let-
ters are significantly different (p < 0.05). Abbreviation of compounds is
shown in the abbreviation list.
corresponding glycosides.20) The hydrophobicity of per-
oxisome proliferators was found to be positively corre-
lated with their binding affinity to mouse peroxisome
proliferator-activated receptor (mPPARα).21) Therefore,
it is plausible that in our study the hydrophobicity of
RA and related compounds may have affected their
interaction with a cross-linked IgE/FcεRI receptor8) at
the cell surface, inhibiting the signaling events leading
to degranulation of RBL-2H3 cells. LogP values were
calculated (cLogP in Table 1) using the software,
Molinspiration Cheminformatics 2014 (http://www.mol
inspiration.com/cgi-bin/properties), and plotted against
pIC50 (−logIC50) values for β-hexosaminidase release
(Fig. 2). For the RA ester series, as cLogP increased
from 1.6 (RA) to 4.7 (RA-hex), β-hexosaminidase
release suppression was enhanced from IC50 value of
1313.7 μM (RA) to 23.7 μM (RA-pro), and then
decreased to 34.5 μM (RA-hex) (Table 1). For CA and
Cou ester series, a similar tendency of β-hexosamini-
dase release suppression was observed to that of RA
ester series. Consequently, strong quadratic correlations
were observed between cLogP and pIC50 for each of
Another point that should be emphasized in this
experiment is the dramatic decrease of antiallergic
activity of CA-hex (IC50 523 μM) and Cou-hex (IC50
1163 μM) compared with the mild decrease of activity
of the RA-hex (IC50 35 μM) against the propyl ester.
The IC50 values of the 5 methyl esters, RA-me,

4
F. Zhu et al.
Fig. 2. The relationship between calculated octanol–water partition coefficient (cLogP) and β-hexosaminidase release suppression (pIC50) of RA,
CA, p-coumaric acid, and their esters.
Note: Abbreviation of compounds shown here was followed the abbreviation list.
CA-me, Cou-me, FA-me, and Cin-me were 57 μM,
120 μM, 182 μM, 144 μM, and 334 μM, respectively.
RA-me, CA-me, and FA-me exhibited greater suppres-
sion activity than the other two methyl esters, indicat-
ing that the binding affinity of those compounds to
binding sites depends on the number of hydroxyl
groups and methoxy groups on the backbone aromatic
rings. The methyl esterification of these compounds
dramatically improved the suppression effect (Fig. 1),
and that could be due to the increased hydrophobicity
as well as the negligible steric hindrance of short alkyl
chains when binding to the IgE/FcεRI receptor. We
suggest that the increase of bulkiness of alkyl groups
could have interfered with the binding affinity between
the binding sites and esters with long alkyl chain, such
as CA-hex (IC50 523 μM) and Cou-hex (IC50
1163 μM). However, the higher number of hydroxyl
groups, as mentioned above, in the RA backbone may
imply a stronger affinity (e.g. by hydrogen bonds) for
the binding sites, which may be less affected by the
steric hindrance caused by the bulky hexyl groups.
Molecular docking experiments are necessary to clarify
this mechanism, and will be done in the near future.
Furthermore, the binding affinity of RA-me and CA-
me, bearing 3,4-dihydroxyphenyl group(s), to the
cross-linked IgE/FcεRI receptor may be stronger than
those of Cou-me, FA-me, and Cin-me. The 3,4-dihy-
droxyphenyl moieties of RA- and CA- free/ester series
could be essential for the binding of these acids/esters
and the cross-linked IgE/FcεRI receptor. The higher
inhibitory activity of RA-esters (possessing two
3,4-dihydroxyphenyl moieties) compared with that of
CA-esters (possessing only one 3,4-dihydroxyphenyl
moiety) support this hypothesis.
Furthermore, our data indicates that the 3,4-dihydroxy-
phenyl moieties in phenolic ester backbone could be
essential for the inhibitor–binding site interaction. The
elucidation of mechanism underlying the interactions
between phenolic acids/esters warrants further investi-
gations.
Funding
This study was supported by Monbukagakusho Scholarship
(Ministry of Education, Culture, Sports, Science and Technol-
ogy, Japan).
References
[1] Jun H-I, Kim B-T, Song G-S, Kim Y-S. Structural characteriza-
tion of phenolic antioxidants from purple perilla (Perilla frutes-
cens var. acuta) leaves. Food Chem. 2014;148:367–372.
[2] Oh H, Park C-S, Ahn HJ, Park YS, Kim HM. Effect of Perilla
frutescens var. acuta Kudo and rosmarinic acid on allergic
inflammatory reactions. Exp. Biol. Med. 2011;236:99–106.
[3] Osakabe N, Yasuda A, Natsume M, Sanbongi C, Kato Y, Osawa
T, Yoshikawa T. Rosmarinic acid,
a major polyphenolic
component of Perilla frutescens, reduces lipopolysaccharide
(LPS)-induced liver injury in D-galactosamine (D-GalN)-sensi-
tized mice. Free Radical Biol. Med. 2002;33:798–806.
[4] Sanbongi C, Takano H, Osakabe N, Sasa N, Natsume M,
Yanagisawa R, Inoue K, Sadakane K, Ichinose T, Yoshikawa T.
Rosmarinic acid in perilla extract inhibits allergic inflammation
induced by mite allergen, in a mouse model. Clin. Exp. Allergy.
2004;34:971–977.
[5] Osakabe N, Takano H, Sanbongi C, Yasuda A, Yanagisawa R,
Inoue K, Yoshikawa T. Anti-inflammatory and anti-allergic effect
of rosmarinic acid (RA); inhibition of seasonal allergic rhinocon-
junctivitis (SAR) and its mechanism. BioFactors. 2004;21:
127–131.
[6] Takano H, Osakabe N, Sanbongi C, Yanagisawa R, Inoue K,
Yasuda A, Natsume M, Baba S, Ichiishi E-I, and Yoshikawa T.
Extract of Perilla frutescens enriched for rosmarinic acid, a poly-
phenolic phytochemical, inhibits seasonal allergic rhinoconjuncti-
vitis in humans. Exp. Biol. Med. 2004;229:247–254.
[7] Abramson J, Licht A, Pecht I. Selective inhibition of the
FcɛRI-induced de novo synthesis of mediators by an inhibitory
receptor. EMBO J. 2006;25:323–334.
We conclude that the esterification of rosmarinic,
caffeic, and p-coumaric acids with short alkyl chains
influences their antiallergic activity. Propyl esters
exhibited the highest antiallergic activity. Hydrophobic-
ity and steric bulkiness of the alkyl chain are likely to
be modulating the interaction between phenolic acid/
esters, and the binding sites in the IgE/FcεRI receptor.

Rosmarinic acid esters contribute to antiallergic activity
5
[8] Abramson J, Pecht I. Regulation of the mast cell response to the
type I Fcε receptor. Immunol. Rev. 2007;217:231–254.
[9] Pecht I, Corcia A. Stimulus-secretion coupling mechanisms in
mast cells. Biophys. Chem. 1987;26:291–301.
[17] Zhu F, Asada T, Sato A, Koi Y, Nishiwaki H, Tamura H.
Rosmarinic acid extract for antioxidant, antiallergic, and α-gluco-
sidase inhibitory activities, isolated by supramolecular technique
and solvent extraction from perilla leaves. J. Agric. Food. Chem.
2014;62:885–892.
[18] Shimoda H, Tanaka J, Yamada E, Morikawa T, Kasajima N,
Yoshikawa M. Anti type I S of Japanese butterbur extract and
its mast cell degranulation inhibitory ingredients. J. Agric. Food.
Chem. 2006;54:2915–2920.
[19] Sato A, Shinozaki N, Tamura H. Secoiridoid type of antiallergic
substances in olive waste materials of three Japanese varieties of
Olea europaea. J. Agric. Food. Chem. 2014;62:7787–7795.
[20] Morito K, Hirose T, Kinjo J, Hirakawa T, Okawa M, Nohara T,
Ogawa S, Inoue S, Muramatsu M, Masamune Y. Interaction of
phytoestrogens with estrogen receptors alpha and beta. Biol.
Pharm. Bull. 2001;24:351–356.
[21] Lewis DFV, Lake BG. Quantitative structure-activity relationship
(QSAR) analysis for a series of rodent peroxisome proliferators:
interaction with the mouse liver peroxisome proliferator-activated
receptor α (mPPARα). Toxicol. In Vitro. 1997;11:99–105.
[22] Hlavacek WS, Posner RG, Perelson AS. Steric effects on multi-
valent ligand-receptor binding: exclusion of ligand sites by
bound cell surface receptors. Biophys. J. 1999;76:3031–3043.
[23] Holowka D, Sil D, Torigoe C, Baird B. Insights into immuno-
globulin E receptor signaling from structurally defined ligands.
Immunol. Rev. 2007;217:269–279.
[10] Aketani S, Teshima R, Umezawa Y, Sawada J. Correlation
between cytosolic calcium concentration and degranulation in
RBL-2H3 cells in the presence of various concentrations of
antigen-specific IgEs. Immunol. Lett. 2001;75:185–189.
[11] Huang F, Yamaki K, Tong X, Fu L, Zhang R, Cai Y,
Yanagisawa R, Inoue K, Takano H, Yoshino S. Inhibition of the
antigen-induced activation of RBL-2H3 cells by sinomenine. Int.
Immunopharmacol. 2008;8:502–507.
[12] Tokura T, Nakano N, Ito T, Matsuda H, Nagasako-Akazome Y,
Kanda T, Ikeda M, Okumura K, Ogawa H, Nishiyama N. Inhibi-
tory effect of polyphenol-enriched apple extracts on mast cell
degranulation in vitro targeting the binding between IgE and
FcεRI. J. Agric. Food. Chem. 2008;56:12073–12080.
[13] Li Y, Lee S-H, Le Q-T, Kim M-M, Kim S-K. Anti-allergic
effects of phlorotannins on histamine release via binding inhibi-
tion between IgE and FcεRI. J. Agric. Food. Chem.
2008;56:12073–12080.
[14] Handlogten MW, Kiziltepe T, Moustakas DT, Bilgiçer B.
Design of a heterobivalent ligand to inhibit IgE clustering on
mast cells. Chem. Biol. 2011;18:1179–1188.
[15] Chung SY, Champagne ET. Reducing the allergenic capacity of
peanut extracts and liquid peanut butter by phenolic compounds.
Food Chem. 2009;115:1345–1349.
[16] Yamashita S, Tsukamoto S, Kumazoe M, Kim Y, Yamada K,
Tachibana H. Isoflavones suppress the expression of the FcεRI
high-affinity immunoglobulin E receptor independent of the
estrogen receptor. J. Agric. Food. Chem. 2012;60:8379–8385.
[24] Ahmed S, James K, Owen CP, Patel CK, Patel M. Hydrophobic-
ity, a physicochemical factor in the inhibition of the enzyme
estrone sulfatase (ES). Bioorg. Med. Chem. Lett. 2001;11:
2525–2528.