Xantone Amilaza Si Glucozidaza
Xantone Amilaza Si Glucozidaza
Xantone Amilaza Si Glucozidaza
com
Abstract—We have proven that xanthones 1–8 isolated from the root of C. tricuspidata possess highly potent aa-glucosidase inhi-
bition properties. Compound 1 was identified as a new isoprenylated tetrahydroxy xanthone, 1,3,6,7-tetrahydroxy-2-(3-methylbut-2-
enyl)-8-(2-methylbut-3-en-2-yl)-9H-xanthen-9-one (1). These are the first natural xanthones documented to exhibit such inhibition.
The IC50 values of compounds 1–8 inhibiting a-glucosidase activity were determined to be up to 16.2 lM. Mechanistic analysis
showed the xanthones 1–8 exhibited full mixed inhibition.
Crown copyright Ó 2007 Published by Elsevier Ltd. All rights reserved.
a-Glucosidase inhibitors have come to the fore of bio- more dependent on the gp120-CD4 interaction. a-Glu-
medical research into the treatment of numerous dis- cosidase inhibitors have thus reduced the rate of viral
eases including diabetes mellitus type II,1 cancer,2 and proliferation in HIV infection.7 Currently the most
HIV.3 The diverse therapeutic roles of these inhibitors widely used glucosidase inhibitors, such as acarbose
stem from the paramount role played by carbohydrates and voglibose, are iminosugars. Although numerous
in biochemistry.4 For instance: by retarding the cleavage glucosidase inhibitors have been developed, including
of complex carbohydrates, postprandial glucose absorp- chalcones,8 azasugars,9 isoxazoles,10 and aminosugars,11
tion in vivo can be attenuated, thus regulating blood su- these can be tedious to synthesize and are thus not opti-
gar levels in diabetics5; the spread of cancer as well as mal. Recently, certain synthetic xanthones12 have also
the structural changes of cell surface glycoconjugates been identified as a-glucosidase inhibitors, but to date
within neoplasmic cells is proliferated by glycosidases no such inhibitors derived from natural sources have
in the sera and interstitial fluid around the tumor, thus been evaluated.
by effecting glycosidase inhibition, cancer growth may
be retarded6; finally, cellular signaling/recognition is Cundrania tricuspidata is one of the most ubiquitous
principally orchestrated by glycoproteins. In HIV glyco- traditional herbal remedies in East Asia. This plant’s
protein 120 (gp120) is responsible for choreographing beneficial effects have been traditionally associated with
the union of the virus and its CD4+ T-cell targets, via anti-inflammatory, anti-tumor, and anti-gastritis activ-
a specific interaction with CD4. ity. In our recent publications, we have elucidated the
vast biological potential of these species through various
Furthermore, the ensuing syncytium formation (aggre- biological studies.13,14 For instance, xanthones possess-
gation of healthy T cells with the infected cell), which ing a vicinal dihydroxy group on one phenyl ring
grossly accelerates the rate of initial infection, is even were shown to exhibit excellent radical scavenging
activities.15 Interestingly, xanthone-derived species were
Keywords: a-Glucosidase inhibition; Xanthone; Cudrania tricuspidata. later demonstrated to possess potent suppressing
* Corresponding authors. Tel.: +82 55 751 5472; fax: +82 55 757 0178 properties toward LDL oxidation, which we linked to
(K.H.P.); tel.: +82 42 860 4278; fax: +82 42 861 2675 (W.S.L.); e-mail the anti-atherosclerotic and anti-inflammatory nature
addresses: [email protected]; [email protected] of C. tricuspidata.13
0960-894X/$ - see front matter Crown copyright Ó 2007 Published by Elsevier Ltd. All rights reserved.
doi:10.1016/j.bmcl.2007.10.007
6422 E. J. Seo et al. / Bioorg. Med. Chem. Lett. 17 (2007) 6421–6424
In this manuscript, we present our most recent results chemistry to 7. The structure of compound 3 was fully
surveying the a-glucosidase inhibitory ability of a range confirmed with X-ray crystallographic analysis
of xanthones from C. tricuspidata, as defined by their (CCDC-222304) and HMBC correlation in our previous
IC50 values. We hope that this study can elucidate fur- work.16 Thus, compound 1 was identified as 1,2,6,7-tet-
ther the multitude of roles ascribed to this important rahydroxy-2-(3-methylbut-2-enyl)-8-(2-methylbut-3-en-
plant (Fig. 1). 2-yl)-9H-xanthen-9-one.18
The methanol extract of C. tricuspidata showed glycosi- The isolated compounds were tested for their enzymatic
dase inhibitory activity. In our ensuing investigations, inhibitory activities against a-glucosidase from baker’s
we isolated eight xanthones 1–8 from it. The identifica- yeast. The enzyme was assayed according to standard
tion of isolated xanthones 1–8 was performed using procedures by following the hydrolysis of nitrophenyl
2D-NMR together with other spetroscopic data, all glycoside spectrophotometrically.19,20 All compounds
of which was compared with previous work. Com- showed dose-dependent inhibitory effect on a-glucosi-
pounds 2–8 were identified as Macluraxanthone B (2), dase activity (Fig. 2). The inhibitory potencies and
Cudraxanthone L (3), 1,3,7-trihydroxy-4-(1,1-dimethyl- capacities of these polyphenol toward a-glucosidase
2-propenyl)-5,6-(2-2-dimethylchromeno)xanthone (4), activity were investigated.
Cudraxanthone M (5), Cudraxanthone D (6), Cudratri-
cusxanthone F (7), and Cudratricusxanthone A (8).16,17 As shown in Table 1, all xanthones investigated apart
from xanthone 5 exhibited a significant degree of a-glu-
Compound 1 was obtained as yellowish solid having the cosidase inhibition (IC50 16.2–52.9 lM). However the
molecular formula C23H24O6 and 12 degrees of unsatu- activity was significantly affected by subtle changes in
ration from its HREIMS data. The UV and IR spectra
display the properties of 1,3,6,7-tetrahydroxyxanthone
derivatives. The presence of the 1,1-dimethylallyl group 100
was deduced from the connectivity between H-18 and
the vinylic proton H-19a/b (dH 5.34) and the correlation
between C-16, 17 (dC 27.6) and H-18 in the HMBC 80
experiment. This group resided at C-2 because C-15 cor-
Activity (%)
13
11 17
10 O OH O OH O OH O OH
1 19
HO 8a 9a HO HO HO
9
7 2 18
HO 6
5
O 3 OH HO O OH HO O OH O O OH
4
1 2 3 4
O OH O OH
HO HO O OH O OH
HO HO
HO O O HO O O
HO O OCH3 HO O OH
5 6 7 8
1/v (O.D/min)
3 52.9 ± 2.1 7.4
4 24.9 ± 1.1 5.8 80
5 >100 NT
6 32.0 ± 2.7 15.7 60
7 16.2 ± 0.4 7.0
8 37.7 ± 1.8 12.4 40
a
Values are means of three experiments, standard deviation is given in 20
parentheses (NT, not tested).
0
3 -2 -1 0 1 2 3
structure. Perhaps most poignantly, inactive xanthone 5 1/[S, mM]
differed only by the relative position of the 3,3-dimethyl-
allyl appendage (C-4 vs C-5) from one of the most effec- Figure 4. Lineweaver–Burk plots of compound 6 (d, control; .,
tive inhibitors screened, xanthone 6. However, C-4 and 10 lM; ,, 20 lM).
C-8 regioisomeric xanthones 1 and 2 were of almost
equal efficacy, as were regioisomers 1 and 8. Perhaps
most importantly, compound 3, a regioisomer of 1, 2, tion of a-glucosidase by compound 6 is illustrated in
and 8 possessing substitution at C-4, exhibited the Figure 3, representatively. Plots of the initial velocity
second lowest activity. It thus seems that alkyl substitu- versus enzyme concentrations in the presence of different
tion in the 4-position diminishes the potency of the concentrations of compound 6 gave a family of straight
inhibitors greatly, whilst other positions show significant lines, all of which passed through the origin. Increasing
tolerance. the inhibitor concentration resulted in the lowering of
the slope of the line, indicating that these compounds
This was however not the only effect at play in this sys- were reversible inhibitors. We progressed to analyze
tem. C3O-Methylated species 7 was by far the most the mode of inhibition using Lineweaver–Burk plots
effective inhibitor, but its demethylated analog 8 was (Fig. 4), which showed that all xanthones displayed
much less efficient. This implicates H-bond donor ability mixed inhibition.
or polarity as an important facet in defining the activity.
Finally, it has been shown previously in synthetic xant- In conclusion, we have proven that numerous xanthones
hones that large conjugated systems serve as optimal isolated from the root bark of C. tricuspidata possess
inhibitors.12 The potency of the most effective inhibitor highly potent a-glucosidase inhibition properties. These
7 (IC50 = 16.2 lM) compares with sugar-derived gluco- represent the first natural xanthone-derived a-glucosi-
sidase inhibitors currently used for therapeutic pur- dase inhibitors documented.
poses, such as voglibose (IC50 = 23.4 lM)21 and
deoxynojirimycin (IC50 = 3.5 lM).22
Acknowledgments
All inhibitors manifested the same relationship between
enzyme activity and enzyme concentration. The inhibi- This work was financially supported by a grant (Bio-
Green 21 project) from the Rural Development Admin-
istration and the MOST/KOSEF to the Environmental
Biotechnology National Core Research Center (Grant
R15-2003-012-02001-0), Korea. E.J. Seo was supported
0.12 by a grant from the BK21 program.
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