This article was downloaded by: [University of Utah]

On: 28 November 2014, At: 08:20

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

Synthesis and Antioxidative Activity of 3′,4′,6,7-

Tetrahydroxyaurone, a Metabolite of Bidens frondosa
a a a
Somepalli VENKATESWARLU , Gopala K. PANCHAGNULA & Gottumukkala V. SUBBARAJU
a
Laila Impex R & D Centre, Unit I, Phase III, Jawahar Autonagar
Published online: 22 May 2014.

To cite this article: Somepalli VENKATESWARLU, Gopala K. PANCHAGNULA & Gottumukkala V. SUBBARAJU (2004) Synthesis
and Antioxidative Activity of 3′,4′,6,7-Tetrahydroxyaurone, a Metabolite of Bidens frondosa , Bioscience, Biotechnology,
and Biochemistry, 68:10, 2183-2185, DOI: 10.1271/bbb.68.2183

To link to this article: http://dx.doi.org/10.1271/bbb.68.2183

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., 68 (10), 2183–2185, 2004

Note
Synthesis and Antioxidative Activity of 30 ,40 ,6,7-Tetrahydroxyaurone,
a Metabolite of Bidens frondosa*
Somepalli V ENKATESWARLU, Gopala K. P ANCHAGNULA, and Gottumukkala V. S UBBARAJUy
Laila Impex R & D Centre, Unit I, Phase III, Jawahar Autonagar, Vijayawada 520 007, India

Received May 6, 2004; Accepted July 13, 2004

30 ,40 ,6,7-Tetrahydroxyaurone (1a), an aurone isolated
from Bidens frondosa, and five analogues (1b–1f) were
synthesized from pyrogallol in three steps. The antiox-
idative activity of 1a–1f was determined by the super-
oxide free radical and 1,1-diphenyl-2-picrylhydrazyl
(DPPH) free radical scavenging methods.
Downloaded by [University of Utah] at 08:20 28 November 2014
hydes in the presence of an acidic or basic reagent or
neutral alumina.3) 30 ,40 ,6,7-Tetrahydroxyaurone (1a,
maritimetin) has been isolated from Bidens frondosa4)
and other species.5) In view of the importance of dietary
antioxidants in the chemoprevention of such degener-
ative illnesses as cancer, Alzheimer’s, Parkinson’s, and

cardiovascular diseases, we synthesized 1a together with

Key words: 2-[(3,4-Dihydroxyphenyl)methylene]6,7-di-
hydroxybenzo[b]furan-3-one; maritimetin;
synthesis; antioxidative activity
Aurones, 2-benzylidenebenzofuran-3(2H)-ones, are
naturally occurring yellow pigments of plants that are
structurally related to flavonoids.1) Aurones have limited
natural occurrence and are generally synthesized from 2-
hydroxychalcones by using mercury(II) acetate,2) or by
condensation between benzofuranones and benzalde-
five new structural analogs, 1b–1f, for the first time and
evaluated their antioxidative activity.
The Friedal–Crafts reaction of 2 with chloroacetic
acid in the presence of BF3 diethyl etherate gave 3
which, upon cyclization in the presence of sodium
acetate as a base, yielded 6,7-dihydroxycoumaranone (4)
in an 85% yield. Acid-catalyzed condensation of 4 with
3,4-dihydroxybenzaldehyde afforded aurone (1a) in a
40% yield (Scheme 1). Synthetic 1a was supported by
the spectral data (UV, IR, 1 H NMR, 13 C NMR and

OH OH OH

HO OH HO OH HO O
(i) (ii)

Cl
2 4 O
O
3

R4
5'
OH 4' R3
6'
HO 6 7
(iii) O1 1' 3'
a = R1=R4=H, R2=R3=OH
R2
Z 2' b = R1=R4=H, R2=OCH3, R3=OH
5 3
R1 c = R1=R2=R4=H, R3=OH
4
O d = R2=R4=H, R1=R3=OH
e = R2=R3=R4=OH, R1=H
1 f = R1=R2=R4=H, R3=F

Scheme 1. Reagents and conditions: (i) chloroacetic acid, BF3 diethyl etherate, 65
C, 3 h, 55%; (ii) NaOAc, ethanol, reflux, 6 h, 85%; (iii)
substituted benzaldehyde, Ac2 O, 90
C, 2 h, 21–48%.

* Laila Impex Communication # 27.

y
To whom correspondence should be addressed. Tel: +91-866-2541303; Fax: +91-866-2546216; E-mail: subbarajugottumukkala@hotmail.com
Abbreviations: NBT, nitro blue tetrazolium; DPPH, 1,1-diphenyl-2-picrylhydrazyl; BHT, butylated hydroxytoluene; BF3 , boron trifluoride; NaCN,
sodium cyanide; EDTA, ethylenediaminetetraacetic acid
 2184
Table 1. Antioxidative Activity of Aurones 1a–1f
NBT superoxide DPPH radical
S. No Compound scavenging activity scavenging activity
(IC50
M) (IC50
M)
1 1a 6.5 8.3
2 1b 9.0 8.7
3 1c 10.0 10.4
4 1d 12.3 10.1
5 1e 4.3 7.9
6 1f 20.2 11.0
7 BHT 301.8 13.4
8 vitamin C 670.5 23.3
9 vitamin E 530.2 >1000
10 resveratrol 482.5 35.5
mass). To understand the effect of substituents to the
aurone structure on the antioxidative activity, we
synthesized five other analogs, 1b–1f, by the same
method with appropriately substituted benzaldehydes. In
Downloaded by [University of Utah] at 08:20 28 November 2014
all cases, a single geometric isomer (Z) was obtained.
The stereochemistry at the double bond was confirmed
by diagnostic 13 C-NMR data6) (exocyclic olefinic carbon
=CH, resonating at about 111 ppm). It is known that the
reaction proceeds stereoselectively and affords only the
(Z)-aurones. (Z)-Isomer is thermodynamically more
stable than the (E)-isomer.7)
We determined the antioxidative activity of 1a and its
analogs, 1b–1f, by the superoxide free radical scaveng-
ing (NBT) method8) and DPPH method.9) The IC50
values of these compounds are presented in Table 1.
Aurones 1a (IC50 : 6.5
M) and 1e (IC50 : 4.3
M) having
catechol and pyrogallol moieties were the most active
compounds, followed by 1b (IC50 : 9
M), 1c (IC50 :
10
M), 1d (IC50 : 12.3
M) and 1f (IC50 : 20.2
M).
Interestingly 1a and 1e showed several-fold more potent
activity than vitamin C (IC50 : 670
M), vitamin E (IC50 :
530
M), resveratrol (IC50 : 482
M) and BHT (IC50 :
301
M). The same order of activity was followed by
aurones 1a–1f with the DPPH method. Again 1a (IC50 :
8.3
M) and 1e (IC50 : 7.9
M) showed good DPPH free
radical scavenging activity. The superior antioxidative
activity of these compounds lends further support to the
fact that the pyrogallol or catechol system enhances the
antioxidative activity.10)
In conclusion, we synthesized 1a together with the
five analogs, 1b–1f, and evaluated their antioxidative
potential by the two commonly used methods, the
superoxide and DPPH free radical scavenging methods.
Tetrahydroxyaurone (1a) and pentahydroxyaurone (1e)
were both potent antioxidants.
Experimental
See refs. 8 and 9 for the general experimental and
antioxidative activity determination procedures. 1 H-
NMR (400 MHz) and 13 C-NMR (100 MHz) data were
recorded in DMSO-d6 .
General procedure for the preparation of 1a–1f: A
mixture of 4 (3.0 mmol) and substituted benzaldehyde
S. VENKATESWARLU et al.
(3.0 mmol) in acetic anhydride (7.5 ml) was heated at
90
C for 2 h. The cooled reaction mixture was poured
into ice-cooled water, extracted with diethyl ether, and
the solvent was removed. The residue was dissolved in
methanol (7.5 ml) and HCl (20%, 7.5 ml) and then
refluxed for 1 h. The cooled reaction mixture was diluted
with ice-cooled water and extracted with ethyl acetate.
The residue obtained after evaporating the solvent was
chromatographed in a silica gel column, using chloro-
form-methanol (90:10) as the eluent, to give aurones
1a–1f.
2-[(3,4-Dihydroxyphenyl)methylene]6,7-dihydroxyben-
zo[b]furan-3-one (1a): mp 286–288
C (lit.11) mp 280–
292
C); UV max (MeOH) nm ("): 205 (63,860), 411
(53,180); IR max (KBr) cm1 : 3366, 1604, 1290, 1192,
1125, 1036; NMR H : 6.63 (1H, s, =CH), 6.74 (1H, d,
J ¼ 8:3 Hz, H-5), 6.86 (1H, d, J ¼ 8:2 Hz, H-50 ), 7.13
(1H, d, J ¼ 8:3 Hz, H-4), 7.38 (1H, dd, J ¼ 8:4 &
1.8 Hz, H-60 ), 7.44 (1H, d, J ¼ 1:9 Hz, H-20 ), 9.1–9.8
(3H, brs, 3  Ar–OH), 10.65 (1H, brs, Ar–OH); NMR
C : 182.0, 155.1, 154.2, 147.9, 145.9, 145.4, 130.1,
124.6, 123.6, 118.4, 116.0, 115.2, 114.6, 112.6, 111.8;
MS (ESI, negative scan): m=z 285 (M  H) .
2-[(4-Hydroxy-3-methoxyphenyl)methylene]6,7-dihy-
droxybenzo[b]furan-3-one (1b): mp 264–266
C; IR
max (KBr) cm1 : 3518, 3361, 1669, 1278, 1198, 1150,
1034; NMR H : 3.86 (3H, s, –OCH3 ), 6.74 (1H, s,
=CH), 6.75 (1H, d, J ¼ 8:2 Hz, H-5), 6.92 (1H, d,
J ¼ 8:0 Hz, H-50 ), 7.13 (1H, d, J ¼ 8:2 Hz, H-4), 7.50–
7.60 (2H, m, H-20 , 60 ), 9.72 (2H, brs, 2  Ar–OH), 10.61
(1H, brs, Ar–OH); NMR C : 182.0, 155.1, 154.4, 148.8,
147.7, 146.1, 130.0, 125.7, 123.6, 116.1, 115.5, 115.4,
114.5, 112.7, 111.6, 55.8; MS (ESI, negative scan): m=z
299 (M  H) . Elemental analysis. Found: C, 63.78; H,
3.98%. Calcd. for C16 H12 O6 : C, 64.00; H, 4.03%.
2-[(4-Hydroxyphenyl)methylene]6,7-dihydroxybenzo-
[b]furan-3-one (1c): mp 296–299
C; IR max (KBr)
cm1 : 3400, 1678, 1631, 1297, 1251, 1162, 1041; NMR
H : 6.72 (1H, s, =CH), 6.73 (1H, d, J ¼ 8:3 Hz, H-5),
6.90 (2H, d, J ¼ 8:6 Hz, H-30 ,50 ), 7.13 (1H, d, J ¼
8:3 Hz, H-4), 7.92 (2H, d, J ¼ 8:6 Hz, H-10 ,60 ), 9.80–
10.40 (3H, br s, 3  Ar–OH); NMR C : 182.0, 159.1,
155.0, 154.2, 145.9, 133.4, 130.0, 123.2, 115.9, 115.2,
114.5, 112.7, 111.2; MS (ESI, negative scan): m=z 269
(M  H) . Elemental analysis. Found: C, 66.29; H,
3.67%. Calcd. for C15 H10 O5 : C, 66.67; H, 3.73%.
2-[(2,4-Dihydroxyphenyl)methylene]6,7-dihydroxyben-
zo[b]furan-3-one (1d): mp 226–229
C; IR max (KBr)
cm1 : 3233, 1655, 1615, 1283, 1159, 1095, 1042; NMR
H : 6.41 (1H, s, =CH), 6.42 (1H, d, J ¼ 8:8 Hz, H-50 ),
6.73 (1H, d, J ¼ 8:3 Hz, H-5), 7.07 (1H, s, H-30 ), 7.10
(1H, d, J ¼ 8:3 Hz, H-4), 8.16 (1H, d, J ¼ 8:8 Hz, H-60 ),
9.49 (1H, brs, Ar–OH), 9.98 (1H, brs, Ar–OH), 10.24
(1H, brs, Ar–OH), 10.50 (1H, brs, Ar–OH); NMR C :
182.0, 160.8, 159.1, 154.8, 153.9, 145.4, 133.0, 130.1,
115.1, 114.9, 112.6, 110.8, 108.3, 105.9, 102.3; MS
(ESI, negative scan): m=z 285 (M  H) . Elemental
analysis. Found: C, 62.67; H, 3.47%. Calcd. for
 Synthesis and Antioxidative Activity of 30 ,40 ,6,7-Tetrahydroxyaurone
C15 H10 O6 : C, 62.94; H, 3.52%.
2-[(3,4,5-Trihydroxyphenyl)methylene]6,7-dihydroxy-
benzo[b]furan-3-one (1e): mp 294–296
C; IR max
(KBr) cm1 : 3313, 1671, 1605, 1201, 1134, 1042;
NMR H : 6.52 (1H, s, =CH), 6.74 (1H, d, J ¼ 8:3 Hz,
H-5), 6.99 (2H, s, H-20 ,60 ), 7.12 (1H, d, J ¼ 8:3 Hz, H-
4), 9.00–9.40 (5H, br s, 5  Ar–OH); NMR C : 182.1,
155.1, 154.4, 146.1, 136.4, 130.3, 122.4, 115.3, 114.6,
112.7, 112.2, 111.2; MS (ESI, negative scan): m=z 301
(M  H) . Elemental analysis. Found: C, 59.46; H,
3.29%. Calcd. for C15 H10 O7 : C, 59.61; H, 3.34%.
2-[(4-Fluorophenyl)methylene]6,7-dihydroxybenzo[b]-
furan-3-one (1f): mp 290–293
C; IR max (KBr) cm1 :
3525, 3057, 1655, 1229, 1164, 1132, 1045; NMR H :
6.76 (1H, d, J ¼ 8:3 Hz, H-5), 6.83 (1H, s, =CH), 7.16
(1H, d, J ¼ 8:3 Hz, H-4), 7.36 (2H, dd, J ¼ 8:8 Hz, H-
20 ,60 ), 8.12 (2H, dd, J ¼ 8:7 & 5.7 Hz, H-30 ,50 ), 9.50–
10.40 (2H, br s, 2  Ar–OH); MS (ESI, negative scan):
m=z 271 (M  H) . Elemental analysis. Found: C, 65.83;
Downloaded by [University of Utah] at 08:20 28 November 2014
H, 3.29%. Calcd. for C15 H9 FO4 : C, 66.18; H, 3.33%.
Acknowledgments
We thank Sri G. Ganga Raju, Chairman, and Mr. G.
Rama Raju, Director, of Laila Impex for encouragement,
and Mr. A. V. Krishnaraju for his help in screening
compounds for antioxidant data.
References
1) Harborne, J. B., ‘‘The Flavonoids’’, Chapman and Hall,
London, pp. 340–342 (1988).
2185
2) Sekizaki, H., Synthesis of 2-benzylidene-3(2H)-benzo-
furan-3-ones (aurones) by oxidation of 20 -hydroxychal-
cones with mercury(II) acetate. Bull. Chem. Soc. Jpn.,
61, 1407–1409 (1988).
3) Varma, R. S., and Varma, M., Alumina-mediated
condensation. A simple synthesis of aurones. Tetrahe-
dron Lett., 33, 5937–5940 (1992).
4) Romussi, G., and Pagani, F., Constituents of Bidens
frondosa. Boll. Chim. Farm., 109, 467–475 (1970).
5) Shimokoriyama, M., and Geissman, T. A., Anthochlor
pigments. XIV. The pigments of Viguiera multiflora
(Nutt.) and Baeria chrysostoma (F. and M.). J. Org.
Chem., 25, 1956–1959 (1960).
6) Pelter, A., and Ward, R. S., Carbon-13 nuclear magnetic
resonance spectra of (Z)- and (E)-aurones. J. Chem. Soc.
Perkin Trans. 1, 328–329 (1979).
7) Hastings, J. S., and Heller, H. G., The stereochemistry of
aurones [2-substituted benzylidenebenzofuran-3(2H)-
ones]. J. Chem. Soc. Perkin Trans. 1, 2128–2132 (1972).
8) Venkateswarlu, S., Satyanarayana, B., Sureshbabu, C.
V., and Subbaraju, G. V., Synthesis and antioxidant
activity of 4-[2-(3,5-dimethoxyphenyl)ethenyl]-1,2-ben-
zenediol. Biosci. Biotechnol. Biochem., 67, 2463–2466
(2003).
9) Lamaison, J. I., Ptitjean-Freytet, C., and Carnet, A.,
DPPH scavenging activity. Pharm. Acta Helv., 66, 185–
188 (1991).
10) Wang, M., Jin, Y., and Ho, C.-T., Evaluation of
resveratrol derivatives as potential antioxidants and
identification of a reaction product of resveratrol and
2,2-diphenyl-1-picryhydrazyl radical. J. Agric. Food
Chem., 47, 3974–3977 (1999).
11) Shimokoriyama, M., Anthochlor pigments of Coreopsis
tinctoria. J. Am. Chem. Soc., 79, 214–220 (1957).