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Inhibition of pancreatic lipase by berberine and dihydroberberine: An investigation by docking simulation and experimental validation

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Berberine (BBR) and dihydroberberine (HBBR) were investigated as inhibitors of pancreatic lipase in an attempt to explore their potential hypolipidemic activities. The study included docking simulations and in vitro enzymatic inhibition assays. At the molecular level, docking simulations revealed several significant binding interactions between the docked natural compounds and the key amino acids in the binding pocket of the pancreatic lipase enzyme. BBR had similar pattern of binding inter-actions as HBBR; however, BBR has a permanent cationic center which is suggested to have an adverse influence on ligand–pancreatic lipase affinity. This trend is explainable by the proposition that ionized ligands favor hydration instead of docking into the binding site. This might explain the lower inhibitory activity of BBR comparing to HBBR, which appeared from their estimated IC 50 values. The logarithmic regression of PL inhibition versus concentra-tion revealed estimated IC 50 values of 106 and 8.0 lg/mL for BBR and HBBR, respectively.
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ORIGINAL RESEARCH
Inhibition of pancreatic lipase by berberine
and dihydroberberine: an investigation by docking simulation
and experimental validation
Mohammad Mohammad
Ihab M. Al-masri
Ala Issa
Ayman Khdair
Yasser Bustanji
Received: 7 June 2012 / Accepted: 24 August 2012 / Published online: 6 September 2012
Ó Springer Science+Business Media, LLC 2012
Abstract Berberine (BBR) and dihydroberberine
(HBBR) were investigated as inhibitors of pancreatic lipase
in an attempt to explore their potential hypolipidemic
activities. The study included docking simulations and in
vitro enzymatic inhibition assays. At the molecular level,
docking simulations revealed several significant binding
interactions between the docked natural compounds and
the key amino acids in the binding pocket of the pancreatic
lipase enzyme. BBR had similar pattern of binding inter-
actions as HBBR; however, BBR has a permanent cationic
center which is suggested to have an adverse influence on
ligand–pancreatic lipase affinity. This trend is explainable
by the proposition that ionized ligands favor hydration
instead of docking into the binding site. This might explain
the lower inhibitory activity of BBR comparing to HBBR,
which appeared from their estimated IC
50
values. The
logarithmic regression of PL inhibition versus concentra-
tion revealed estimated IC
50
values of 106 and 8.0 lg/mL
for BBR and HBBR, respectively.
Keywords Berberine Dihydroberberine
Pancreatic lipase Obesity Docking simulations
Enzyme inhibition
Introduction
Obesity is one of the most common nutritional dilemmas in
the modern countries and is considered a potential risk
factor for the development of a plethora of devastating
diseases, including insulin resistance and type 2 diabetes,
lipid profile disorders, osteoarthritis, hyperuricemia,
malignancies, and cardiovascular diseases which includes
hypertension, coronary heart diseases, and stroke (Arbeeny,
2004;Cairns,2005; Gurevich-Panigrahi et al., 2009).
Many recent studies have placed obesity as one of the
greatest threats to global health in this millennium. Recent
reports showed that there are more than one billion over-
weight adults worldwide including at least 300 million
classified as clinically obese with increased risk of mor-
bidity and mortality (Arbeeny, 2004; Vega, 2001). Unfor-
tunately, these numbers are growing in alarming rates.
Obesity is considered as a metabolic disorder, which is
mainly caused by an imbalance between the energy intake
and expenditure. However, comprehensive understanding
of the molecular mechanisms that tightly regulate body
weight has afforded potential opportunities for therapeutic
managements and offered renewed hope for introducing
antiobesity drugs into the market (Bray and Tartaglia,
2000; Foster-Schubert and Cummings, 2006; Halford,
2006b).
Despite the surfeit of research data available on obesity,
it still remains alarming, unsolved medical and socioeco-
nomical problem (Cooke and Bloom, 2006; Halford,
2006a; Hofbauer, 2002). The relatively few medications
that are available in the market for obesity management
have minimal efficacy and are poorly tolerated. As such,
there is an urgent need for the development of new thera-
peutic agents to overcome obesity and its malicious clinical
consequences (Shi and Burn, 2004).
M. Mohammad (&) A. Issa Y. Bustanji
Faculty of Pharmacy, University of Jordan,
Amman 11942, Jordan
e-mail: mkmohammad@ju.edu.jo
I. M. Al-masri
Faculty of Pharmacy, Al-Azhar University,
Gaza, Gaza Strip, Palestine
A. Khdair
Faculty of Pharmacy, Applied Science University,
Amman, Jordan
123
Med Chem Res (2013) 22:2273–2278
DOI 10.1007/s00044-012-0221-9
MEDICINAL
CHEMISTR
Y
RESEARCH
Recently, new approaches for fighting obesity have
involved inhibition of dietary triglyceride absorption
through inhibition of pancreatic lipase (PL) which is being
the major source of excess calories (Ballinger and Peikin,
2002). PL (triacylgycerol acyl hydrolase), the primary
lipolytic enzyme synthesized and released in the pancreatic
juice by the pancreas, plays an essential role in the efficient
digestion and absorption of triglycerides from the gut.
PL is responsible for the hydrolysis and absorption of
50–70 % of total dietary fats.
X-ray crystallography has clarified the three-dimen-
sional structure of human PL. This enzyme is a monomeric
glycoprotein, composed of 449 amino acids, which is
divided into two main folding domains, the larger N-ter-
minal domain and a C-terminal domain (Vantilbeurgh
et al., 1992; Winkler et al., 1990). The N-terminal domain
is the catalytic domain to which substrate binds; while the
C-terminal domain binds to colipase, another enzyme that
acts as a required cofactor for activity (Vantilbeurgh et al.,
1992, Winkler et al., 1990).
Orlistat (Xenical
Ò
), widely prescribed for the manage-
ment of obesity, is the only FDA approved drug that acts
through inhibition of PL. Although Xenical
Ò
is one of the
best-selling drugs in the western countries, its use is
accompanied by uncomfortable intestinal adverse events
like oily stools, oily spotting, and flatulence (Drent et al.,
1995). Natural compounds identified from traditional
medicinal plants provide an enormous pool of PL inhibitors
that can possibly be turned into clinical products (Birari
and Bhutani, 2007; Bustanji et al., 2010, 2011a, b; Issa
et al., 2011).
Berberine (BBR) is an isoquinoline alkaloid found in
Hydrastis canadensis, Berberis, and Cortex phellodendri.It
has been comprehensively reviewed in the literature as an
attractive natural compound of broad potential medical
applications including hypolipidemic effects (Vuddanda
et al., 2010). BBR was found to ameliorate diabetes and
serum lipid profile by reducing serum cholesterol, triglyc-
erides, and LDL-cholesterol in animals and human (Bustanji
et al., 2006; Hu and Davies, 2010;Konget al., 2004;Lee
et al., 2006).
Consequently, current work was devoted to evaluate
BBR’s potential PL inhibitory activity as well as its bio-
logically available derivative dihydroberberine (HBBR).
Initially, we used computer-aided molecular docking of
BBR and HBBR into the binding pocket of PL to reach
preliminary conclusions about BBR/PL and HBBR/PL
binding energetics. Eventually, the docked active com-
pounds were tested in vitro against PL to evaluate their
inhibition potential.
Our findings help in understanding the mechanism of
action of BBR as hypolipidemic agent. It can also provide
new insights toward the binding interactions of BBR and
HBBR with PL which can lead to the screening and dis-
covery of new natural and safe PL inhibitors.
Materials and methods
Materials
Chemicals and reagents used in these experiments were
reagent grade and obtained from the following sources:
porcine PL type II, Tris–HCl buffer, orlistat; BBR, and
p-nitrophenyl butyrate (Sigma, USA). HBBR (PhytoLab
GmbH, Germany).
Docking experiment
Docking simulations and settings were employed as pre-
viously described in Bustanji et al. (2011a). Briefly, the
chemical structure of BBR and HBBR (Fig. 1) were sket-
ched in Chemdraw Ultra (7.01) and saved in MDL molfile
format. Subsequently, an ensemble of energetically acces-
sible conformers were created using OMEGA2 software
(OMEGA, 2006). OMEGA builds initial models of struc-
tures by assembling fragment templates along sigma bonds.
Once an initial model of a structure is built, OMEGA
generates additional models by enumerating ring confor-
mations and invertible nitrogen atoms. The produced
conformers are saved in SD format.
The 3D coordinates of PL were recovered from the Pro-
tein Data Bank (PDB code: 1LPB, resolution; 2.46 (A
˚
)
(Egloff et al., 1995). Hydrogen atoms were inserted to the
protein using the DS visualizer templates for protein resi-
dues. The docking study was performed in the presence of
crystallographically explicit water molecules. The tested
compounds were docked into the binding site of PL using
FRED software (FRED,
2006). The ligand conformers and
protein structure are treated as rigid items during the docking
process. FRED’s docking plan is to comprehensively score
all possible positions of each ligand in the binding pocket
(FRED, 2006). The conformational ensemble of the BBR
N
O
O
H
3
CO
OCH
3
N
O
O
H
3
CO
OCH
3
Berberine Dihydroberberine
Fig. 1 Two-dimensional structure of berberine (BBR) and dihydrob-
erberine (HBBR)
2274 Med Chem Res (2013) 22:2273–2278
123
and HBBR generated using OMEGA software was used as
input in the FRED software. For further details about FRED
parameters which were employed in docking simulations
see reference (Bustanji et al., 2011a).
Preparation of compounds and extract for in vitro assay
BBR or HBBR were initially dissolved in sufficient vol-
umes of DMSO to give five stock solutions with an esca-
lating concentration range of about 5–250 lg/mL.
Thereafter, 20 lL of each stock solution was accurately
transferred to the reaction mixture to produce BBR or
HBBR final concentration range of 0.1–5 lg/mL.
PL inhibition assay
The PL activity was determined using a colorimetric assay
approach as previously described, in which, the release of
p-nitrophenol (PNPB) will be measured spectrophotomet-
rically at 410 nm (Bustanji et al., 2011a). Briefly, 50 mg
(2,000 unit) of crude porcine PL type II (Sigma, USA, EC
3.1.1.3) was suspended in 10 mL of Tris–HCl buffer
(2.5 mmol, pH 7.4 with 2.5 mmol NaCl) and mixed well
using a stirrer for 15 min. Thereafter, the prepared solution
was centrifuged at 1,5009g for 10 min and the clear
supernatant was collected. The final enzyme concentration
was about 200 unit/mL.
BBR and HBBR solutions were preincubated with
0.10 mL of PL solution in Tris–HCl buffer for 5 min at
37 °C. After that, the PNPB substrate (10 mM in acetoni-
trile, 5 lL) was added to obtain a final volume of 1 mL of
the reaction mixture. The absorbance was spectrophoto-
metrically measured at 410 nm for at least 5 time points:
1–5 min. The release of PNPB is estimated as the incre-
ment increase in absorbance against blank. The percentage
of residual activity of PL was determined for BBR and
HBBR by comparing the lipase activity in the presence and
the absence of the tested inhibitors. Orlistat was used as a
positive standard inhibitor and all experiments were repe-
ated thrice.
Results and discussion
Lipid metabolism is cunningly balanced to maintain
homeostasis (Foster-Schubert and Cummings, 2006;
Mukherjee, 2003). When the balance is misplaced, obesity
or dyslipidemia appears ending with a range of serious
metabolic diseases. One of the most important strategies to
control obesity is to develop inhibitors of PL.
Phytochemicals identified from traditional medicinal
plants provide exciting opportunity for the development of
novel antiobesity therapeutics. As a part of our screening
project for biologically active antiobesity agents from
natural resources, BBR and HBBR have been investigated
for their antilipase activities. Several studies, however,
have reported the hypolipidemic activity of BBR alkaloid
(Kong et al., 2004). BBR was found to decrease weight
gain and food intake in mice. Moreover, serum glucose,
triglyceride, and total cholesterol levels were reduced
accompanied with a down-regulation of PPARc expression
in mice fed a high fat diet (Hu and Davies, 2010). Like-
wise, BBR improved lipid profile in obese mice by its
effect on central and peripheral AMP-dependent protein
kinase activities (Kim et al., 2009). Furthermore, oral
administration of BBR in hypercholesterolemic patients
ameliorated lipid profile and reduced serum triglycerides
and LDL-cholesterol with a suggested mechanism of
stimulation of LDL-receptors expression (Kong et al.,
2004).
Our investigation started by evaluating the possibility of
BBR/HBBR-PL binding via computer-aided molecular
modeling technique. Accordingly, both BBR and HBBR
were docked into the binding cleft of PL (PDB code:
1LPB). Generally, docking process consists of two stages:
(i) prediction of the conformation and pose of the bioactive
ligand into the binding pocket, and (ii) evaluation of the
rigidity of target-ligand interactions (scoring). The final
docked conformations are chosen according to their scores.
The docking experiments were performed utilizing the
docking capabilities of the software engine FRED (Fred).
However, an optimal set of parameters for the docking
experiments should be provided to FRED. Therefore, to
identify the optimal docking configuration and scoring
function for PL, the ligand CllP (Fig. 2) was first extracted
from PL crystallographic structure (PDB code: 1LPB)
(Egloff et al., 1995) and then redocked again into the same
protein (self-docking) via a variety of docking conditions.
Chemgauss2 was found to yield the closest model to the
crystallographic structure as shown in Fig. 2.
The preliminary docking study has postulated that BBR
and HBBR might have PL inhibitory activity and this has
encouraged further investigation to evaluate the antilipase
activities of the two alkaloids. Accordingly, BBR and
HBBR were used for in vitro activity bioassay. The enzy-
matic reaction progression was monitored through the
release of PNPB. The in vitro activity was expressed as the
concentration of natural alkaloids that could inhibit PL
activity by 50 % (IC
50
).
Figure 3 shows clearly the dose-dependent relation
between the BBR/HBBR concentrations and the degree of
PL inhibition. However, HBBR showed higher inhibitory
effect (Fig. 3b) than that of BBR (Fig. 3a) as indicated by
Med Chem Res (2013) 22:2273–2278 2275
123
relatively lower IC
50
value of HBBR. The logarithmic
regression of PL inhibition versus concentration revealed
estimated IC
50
values of 106 and 8.0 lg/mL for BBR and
HBBR, respectively (Table 1). Our assay was validated
using orlistat as a positive control with IC
50
equal to
0.21 lg/mL (see Table 1).
On the molecular level, docking simulations have
revealed several significant binding interactions between the
docked natural compounds and the PL (Fig. 4). Comparison
of the docked poses of the BBR and HBBR (Fig. 4b, c) with
the co-crystallized ligand within the binding site of PL
(Fig. 4a) illustrates similarities in their binding profiles.
Both have potential hydrophobic interactions with the key
amino acids Phe-215, Phe-77, and Tyr-114. However, the
dioxymethylene substituted-aromatic ring of BBR and
HBBR is well fitted in the hydrophobic pocket which sug-
gests the existence of van der Waals’ aromatic stacking
attraction between them. Moreover, the two natural com-
pounds having the oxygen of the 1,3-dioxolane ring moiety
seem to be tightly hydrogen-bonded to the hydroxyl and
imidazole NH groups of Ser-152 and His-263, respectively,
albeit at closer proximities in the HBBR (Fig. 4). These
multiple strong hydrogen bonds with Ser-152 and His-263
stabilizes the ligand–protein complex and contributes to the
relatively good affinity of the two alkaloids. Similarly, the
co-crystallized ligand has analogous interactions with Ser-
152 and His-263 (Fig. 4a). In the structure of PL, His-263,
Asp-176, and Ser-152 form a triad representing the lipolytic
site. Furthermore, enzymatic activity has shown to be
diminished after chemical modification of Ser-152 indicat-
ing its essential role for the catalytic activity (Winkler et al.,
1990). Therefore, it is predictable that compounds that
strongly bind to the catalytic triad, especially Ser-152, could
inhibit the lipolytic activity. Although BBR has similar
pattern of binding interactions as HBBR (Fig. 4), it has a
permanent cationic center which may have an adverse effect
on the ligand–PL affinity. This trend is explainable by the
proposition that ionized ligands favor hydration instead of
docking into the binding site, particularly if they are misa-
ligned with their corresponding counterparts in the binding
pocket. A similar analysis was recently used to explain the
general nonspecific enhancement in ligand–receptor affinity
concomitant to increases in ligand lipophilicity (Manly et al.,
2008). This may help to explain the lower affinity of BBR
comparing to HBBR. In our study, the reported significant
decrease in serum cholesterol, triglycerides, LDL-choles-
terol, and weight gain can be attributed, at least partially, to
its inhibitory action of PL. This hypothesis can be supported
by the initial docking studies and in vitro inhibitory assay.
Fig. 2 a Comparison between the docked pose (green) of the ligand
CllP as produced by docking simulation and the crystallographic
structure of this ligand (red) within the binding pocket of PL. b The
solvent accessible surface area of the binding site of PL (1LPB) and
the co-crystallized inhibitor (CllP) [18] (Color figure online)
B
Dihydroberberine
y = 9.5701Ln(x) + 29.991
R
2
= 0.9184
0
20
40
60
80
100
% of Inhibition
A
Berberine
y = 5.0283Ln(x) + 26.526
R
2
= 0.8732
0
20
40
60
80
100
0.01 0.1 1 10
Concentration (µg/mL)
% of Inhibition
0.01 0.1 1 10
Concentration (µg/mL)
Fig. 3 The inhibitory effect of
escalating concentrations of
BBR (a) and HBBR (b) on the
PL activity
Table 1 IC
50
values for tested compounds estimated from logarith-
mic regression of PL inhibition versus concentration
Compound IC
50
(lg/mL)
Berberine (BBR) 106.0
Dihydroberberine (HBBR) 8.0
Orlistat 0.21
2276 Med Chem Res (2013) 22:2273–2278
123
Conclusion
The results of this study clearly proved, through experi-
mental testing and theoretical docking simulations, that
HBBR and BBR have substantial PL inhibition activities.
This PL inhibitory effect can be used to explain, at least
partially, the reported use of BBR as an effective hypo-
lipidemic and fat-mass reduction activities. In conclusion,
this work is considered a step toward the discovery of
new natural and safe hypolipidemic/antiobesity PL
inhibitors.
Acknowledgments This project was sponsored by the Deanship of
Scientific Research at the University of Jordan. The authors wish to
thank the Deanship of Scientific Research at the University of Jordan
for their generous funds. The authors would also like to thank the
OpenEye Scientific Software for providing us with a free license for
FRED software (FRED, version 2.1.5).
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... Based on docking simulations and in vitro enzymatic inhibition assay results, a significant interaction between the docked natural compound and the main amino acids present in the binding pocket of the pancreatic lipase was found. The IC 50 value was also found to be 106 μg/ml, showing promising activity in inhibiting the pancreatic lipase enzyme [56]. ...
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Obesity is a major lifestyle disorder and it is correlated with several ailments. The prevalence of obesity has elevated over the years and it has become a global health problem. The drugs presently used for managing obesity have several side-effects associated with them such as diarrhoea, leakage of oily stools, etc. On the contrary, herbal plants and natural products are considered safe for use because they have lesser side effects. New compounds isolated from medicinal plants are screened and identified to determine their effectiveness and potential in preventing abnormal weight gain. In this review, the medicinal plants and natural materials were surveyed across the literature to cover those that have potential for managing and controlling weight gain, and their mechanism of action, active component, and experimental methodologies are also included. These herbal products can be developed as formulations for therapeutic use in obesity. The herbal plants mentioned in the review are classified based on their mechanism of action: inhibition of pancreatic lipase and appetite suppression activities. The ability to inhibit pancreatic lipase enzyme has been used to determine the effectiveness of herbal products for the prevention of abnormal weight gain because of its action on dietary fat and suppression of appetite. This review is an attempt to summarize the herbal plants and natural products that can be used to develop formulations effective in controlling weight gain and obesity.
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Diabetes is a chronic medical condition due to the lack of insulin or ineffective use of insulin. In recent years, diabetes has become one of the rapidly growing chronic diseases, and it has caused a burden on the individual and the economy. Rhodanine has been identified as an important scaffold in drug discovery because of its wide range of biological properties including antidiabetic. A number of studies have been reported regarding the activity of rhodanine-based compounds in the treatment of diabetes mellitus and its complications. This review aimed to explore the antidiabetic activity of rhodanine-based compounds and their structure–activity relationship (SAR) based on studies that have been reported. Our study discovered several studies about the compounds’ biological screening against a variety of targets, including aldose reductase, α-glucosidase, α-amylase, PPAR-γ and PTPT1B. Some compounds mentioned are also tested in vivo in living organisms. This review offers a resource for future antidiabetic drug development, as many of the hit compounds listed exhibited good antidiabetic properties.
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Pancreatic lipase is a well-known target for obesity drug development. The inhibition of this enzyme mitigates the digestion and absorption of dietary fat. Despite some inconvenient side effects, orlistat is currently the only medication with FDA approval that works by inhibition of pancreatic lipase. Several natural flavonoids, including chalcones, have been reported to possess promising lipase inhibitory activity. In this paper, we describe the evaluation of the lipase inhibitory activity of our in-house natural mimetic chalcone library via virtual screening, followed by the synthesis and biological evaluation of the hits. Compound C82 showed low-micromolar activity against pancreatic lipase in vitro (IC50 = 1.01 ± 0.86 µM). The interaction of C82 and lipase was additionally confirmed by a fluorescence quenching study.
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The ethanolic extract from leaves of Phyllanthus chamaepeuce Ridl. was reported to have the ability to inhibit pancreatic lipase activity in vitro. In order to find the most potent of pancreatic lipase inhibitors from this plant, roots, barks, woods, and leaves which were extracted by using different solvents were used to investigate their ability to inhibit pancreatic lipase activity by in vitro test assay. The findings have shown that ethanolic and methanolic leaf extracts (at a concentration of 100 µg/mL) exhibited the greatest inhibition against porcine pancreatic lipase with 96.13% (IC50: 2.98 ± 1.11 µg/mL) and 95.93% (IC50: 4.36 µg/mL), respectively. Statistical analysis demonstrated strong positive significant correlations between the contents of phytochemical compounds (polyphenols, flavonoids, and alkaloids) and % pancreatic lipase inhibition. The result from enzyme kinetic assay indicated that ethanolic and methanolic leaf extracts from P. chamaepeuce Ridl. noncompetitively inhibited pancreatic lipase activity. These findings seem to indicate the use of P. chamaepeuce Ridl. extracts as an anti-obesity agent.
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The burden of obesity is increasing all over the world. Except for Orlistat, no effective anti-obesity drug is currently available. Therefore, a search for the new anti-obesity compound is need of time. This study demonstrates macromolecular interaction and inhibitory effect of pentacyclic triterpenoids (PTT) on pancreatic lipase (PL). In the present study PTTs from endophytic Colletotrichum gigasporum were found to show significant inhibitory activity against PL with IC50 of 16.62 ± 1.43 μg/mL. The PTT isolated through bioassay-guided isolation showed a dose-dependent (R² = 0.915) inhibition against porcine PL and the results were comparable with the standard (Orlistat). Based on inhibition kinetic data, the gradual increase in Km (app) with increasing PTT concentration indicated that the mode of interaction of PTT with PL was a competitive type, and it directly competed with the substrate (pNPB) for the active site of PL. In vivo studies in Wistar rats at the oral dose (100 mg/kg body weight) of PTT significantly decreased (p < 0.05) incremental plasma triglyceride levels as compared to group B and TG absorption was down-regulated up to 49.18% vis a vis group D animals. The isolated PTT was identified as lupeol based on chromatographic and spectral data. The endophytic isolate was identified as Colletotrichum gigasporum based on morphology and ITS gene sequencing. The present study indicated that PTT had the potential to be used as a natural PL inhibitor in the treatment of obesity and the isolated endophyte can be a valuable bioresource for it.
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Diabesity represents a classic example of a multifactorial disease. The current strategy for the treatment of diabesity is the inhibition of enzymes involved in carbohydrate and lipid metabolism using various drugs. Side effects caused by the drugs available in the market for the treatment of diabesity have necessitated the discovery and development of new novel drugs. In recent years natural molecules have gained importance in the management of multifactorial disease. These molecules are multitargeted in nature and can be useful in the management of diabesity. Some natural molecules are able to regulate gene expression involved in metabolic pathways like adiponectin gene expression, peroxisome proliferator-activated receptor gamma. Apart from gene regulation they are able to inhibit the metabolic enzymes like lipases, amylases and glucosidases. Plants present good sources of natural multifunctional molecules. A number of natural molecules including flavonoids have been reported for multifunctionality. The structural configuration of flavonoids affects the enzymatic activity of lipases and glucosidases which can be useful for designing and development of new drugs for targeting diabesity. The consumption of food in the form of medicine can add a new dimension to the existing drug market. The further acceptance of natural molecules will need to be supported by large clinical studies including studies for toxicity and bioavailability. The present review will provide insight into the anti-diabesity potential of multifunctional natural molecules and will also explain some of the natural molecules in terms of biodistribution and safety. Data from clinical studies of a few such natural molecules is also presented.
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Dihydroberberine (DHBER), the partially reduced form of the alkaloid berberine (BER), is known to exhibit important biological activities. Despite this fact, there have been only few studies that concern the biological properties of functionalized DHBER. Attracted by the potentiality of this latter compound, we have realized the preparation of new arylhydrazono-functionalized DHBERs, starting from BER and some α-bromohydrazones. On the other hand, also the fully reduced form of BER, namely tetrahydroberberine (THBER), and its derivatives have proven to present different biological activities. Therefore, the obtained arylhydrazono-functionalized DHBERs were reduced to the corresponding arylhydrazono-THBERs. The antiproliferative activity of both arylhydrazono-DHBERs and -THBERs has been evaluated on NCI-H1975 lung cancer cells.
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Intense efforts by pharmaceutical industry have been made to identify new targets for obesity-diabetes (Diabesity). Pancreatic triacylglycerol lipase (PL) inhibition is an interesting putative target for obesity management. Fluoroquinolones (FQs) have been identified as potent inhibitors of PL. The aim of this research was to synthesize novel FQs and evaluate their in vitro antilipolytic and antiproliferative properties. Characterization of the synthesized FQs was carried out with NMR, MS, IR and EA. Like orlistat, potential FQs' modulation of PL was quantified colorimetrically (n = 3); further supported by docking studies. Compared to cisplatin, FQs' antiproliferative propensities against a panel of obesity related colorectal cancer cell lines were investigated with Sulforhodamine B assay. Twelve novel FQs (2A-5A, 2B-5B, and 2C-5C) were synthesized and characterized. The PL-IC 50 values of tested FQs were in the range of 6. 8-165.7 µM. FQ 4A was the most active antiproliferative compound against HCT116 with IC 50 value of 3.5 µM. Their selectivity of growth inhibition for safety examination using normal periodontal ligament fibroblasts (PDL) in comparison to cisplatin's lack of differential cytotoxicity was reported. Lipophilicity and hydrogen bonding were found essential for both activities. Conclusively FQs are robustly proven for their emerging in vitro antiobesity and antiproliferative activities.
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Obesity is a main risk factor for cardiovascular, metabolic and endocrine disorders. Despite significant improvements in public education and pharmacologic management in the last two decades, obesity rates have continued to be alarmingly high. The need for a combination of multiple therapy approaches to overcome obesity has become widely accepted by the majority of health care systems and guidelines. The rich potential of nature to combat obesity has not been fully explored yet. Several phytochemicals have been investigated for their potential as lipid lowering agents. We have investigated a total of 23 medicinal plants, belonging to 15 different families and compared their pancreatic lipase inhibitory effects. Inhibition of the pancreatic lipase was chosen as the criteria for therapeutic efficacy since such inhibition would serve two functions. It would provide an adjunctive therapy to the pharmacological agents and would minimize systemic adverse reactions by acting topically in the GI tract. Thirteen plants were found to show in vitro inhibitory activities. The nine most active plants showed an IC;50; range of 107.7-342.7 μg/mL. The plants are Anthemis palaestina Boiss., Salvia spinosa L. Ononis natrix L, Fagonia arabica L., Origanum syriaca L. (Syn. Majorana syriaca (L.) Rafin.), Hypericum triquetrifolium Turra, Malva nicaeensis All., Chrysanthemum coronarium L., Paronychia argentea Lam. Further isolation, identification and characterization of phytoactive compounds responsible for anti-lipase action is required to evaluate the full therapeutic potentials of these plants. © 2011 DAR Publishers/University of Jordan. All Rights Reserved.
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Diabetic dyslipidemia is a key contributor to insulin resistance and a risk factor for cardiovascular complications in patients with DM type 2. Lavandula angustifolia (lavender) is native of the northern region of Jordan and has been long used in Jordanian folkloric medicine in the management of diabetes. To our knowledge, this is the first manuscript to study the potential role of the methanolic extract of the plant in the management of diabetic dyslipidemia. Our results demonstrated in vitro inhibitory effects of L. angustifolia on both hormone sensitive lipase (HSL) and pancreatic lipase (PL) activities. The results indicated that lavender extract inhibited HSL activity in a dose dependent manner with an IC 50 of 175.5 µg/ml. Likewise, it inhibited the PL activity in a dose dependent manner with an IC50 of 56.5 µg/ml. Such inhibitory activities could be attributed, but is not exclusive, to the presence of rosmarinic acid (IC 50 of 125.2 and 51.5 µg/ml for PL and HSL, respectively) and gallic acid (IC 50 of 10.1 and 14.5 µg/ml for PL and HSL, respectively) in the extract. Interestingly, the inhibitory pattern of the lavender on the enzymatic activities of HSL and PL matched the inhibitory pattern of orlistat.
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Diabetic dyslipidemia is a key contributor to insulin resistance and a risk factor for cardiovascular complications in patients with DM type 2. Lavandula angustifolia (lavender) is native of the northern region of Jordan and has been long used in Jordanian folkloric medicine in the management of diabetes. To our knowledge, this is the first manuscript to study the potential role of the methanolic extract of the plant in the management of diabetic dyslipidemia. Our results demonstrated in vitro inhibitory effects of L. angustifolia on both hormone sensitive lipase (HSL) and pancreatic lipase (PL) activities. The results indicated that lavender extract inhibited HSL activity in a dose dependent manner with an IC 50 of 175.5 µg/ml. Likewise, it inhibited the PL activity in a dose dependent manner with an IC50 of 56.5 µg/ml. Such inhibitory activities could be attributed, but is not exclusive, to the presence of rosmarinic acid (IC 50 of 125.2 and 51.5 µg/ml for PL and HSL, respectively) and gallic acid (IC 50 of 10.1 and 14.5 µg/ml for PL and HSL, respectively) in the extract. Interestingly, the inhibitory pattern of the lavender on the enzymatic activities of HSL and PL matched the inhibitory pattern of orlistat.
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Rosmarinus officinalis L. (Rosemary) has been long claimed to have hypogylcemic-hypolipidemic dual effects in folkloric medicine. In an effort to explain rosemary's claimed benefits, numerous published studies have investigated an array of pharmacologic activities of the plant including anti-inflammatory, anticarcinogenic and metabolic effects. The question remained, however, as how rosemary would target both plasma lipids and glucose levels simultaneously. A newer mechanism has been suggested, in which targeting the hormone sensitive lipase (HSL) would be the common link between the two metabolic effects. In fact, HSL has been extensively studied for its effects on the metabolic switch between glucose and free fatty acids (FFAs) as an energy source. The current manuscript summarizes a significant amount of work that was undertaken to identify plant species native to Jordan with potential HSL and pancreatic lipase (PL) inhibitory activities. Our results demonstrated in vitro inhibitory effects of R. officnalis on both HSL and PL in a dose dependent manner. Interestingly, the rosemary extract had an IC 50 for PL that was several fold lower than the IC 50 for HSL, indicating a higher affinity to the former enzyme (13.8 and 95.2 g/mL for PL and HSL, respectively). In addition, we have compared the inhibitory activities of purified constituents found in rosemary to the parent plant [rosmarinic acid (RA), chlorogenic acid (CA), caffeic acid (CaA) and gallic acid (GA)]. Our results showed that all the tested compounds (RA, CA, CaA, and GA) were able to inhibit the PL and HSL activities in a dose dependent manner, but with different potencies. PL and HSL IC 50 values were calculated for each compound and GA was found to be the most potent (IC 50 10.1 and 14.5 for PL and HSL, respectively). Further work is necessary to determine whether our in vitro findings would correlate with the in vivo effects. Nonetheless, our results are a first step in fully understanding the long claimed hypoglycemic-hypolipidemic dual effects of rosemary. Simultaneous targeting of both HSL and PL is likely to open the door for a new era in our continuous battle against DM type 2 and its cardiovascular complications. Currently we are working on identifying the most active constituents of the plant to evaluate a structure-activity relationship which would pave the road for future therapeutic use.
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The prevalence of obesity is increasing at an alarming rate, but, unfortunately, only a few drugs are currently available on the market. In the present study, the methanolic extract of Ginkgo biloba L. (Ginkgoaceae) was investigated as an inhibitor of pancreatic lipase (PL) in an attempt to explain its hypolipidaemic activity. In vitro assay of G. biloba leaves extract revealed a substantial PL inhibition activity (IC(50) = 16.5 µg/mL). Further investigation was performed by employing theoretical docking simulations and experimental testing to uncover the active constituents responsible for G. biloba anti-lipase activity. Virtually, terpene trilactones, including ginkgolides and bilobalide, were found to fit within the binding pocket of PL via several attractive interactions with key amino acids. Experimentally, ginkgolides A, B, and bilobalide were found to inhibit PL significantly (IC(50) = 22.9, 90.0, and 60.1 µg/mL, respectively). Our findings demonstrated that the hypolipidaemic effects of G. biloba extract can be attributed to the inhibition of PL by, at least in part, terpene trilactones. In conclusion, this work can be considered a new step towards the discovery of new natural safe hypolipidaemic PL inhibitors.
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Therapy for transplant rejection, autoimmune disease and allergy must target mature lymphocytes that have escaped censoring during their development. FK506 and cyclosporin are immunosuppressants which block three antigen-receptor signalling pathways (NFAT, NFκB and JNK), through inhibition of calcineurin, and inhibit mature lymphocyte proliferation to antigen. Neither drug induces long-lived tolerance in vivo, however, necessitating chronic use with adverse side effects. Physiological mechanisms of peripheral tolerance to self-antigens provide an opportunity to emulate these processes pharmacologically. Here we use gene-expression arrays to provide a molecular explanation for the loss of mitogenic response in peripheral B-cell anergy, one aspect of immunological tolerance. Self-antigen induces a set of genes that includes negative regulators of signalling and transcription but not genes that promote proliferation. FK506 interferes with calcium-dependent components of the tolerance response and blocks an unexpectedly small fraction of the activation response. Many genes that were not previously connected to self-tolerance are revealed, and our findings provide a molecular fingerprint for the development of improved immunosuppressants that prevent lymphocyte activation without blocking peripheral tolerance.
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The major human lipases include the gastric, pancreatic and bile-salt-stimulated lipase that aid in the digestion and assimilation of dietary fats, and the hepatic, lipoprotein and endothelial lipase that function in the metabolism of lipoproteins. The triacylglycerol and phopholipase activities of these enzymes enable these varied functions. The lipase enzymes exhibit a high degree of sequence homology not only within but also across species. This and the diverse chromosomal location of their genes point to a multigenic family of enzymes involved in absorption and transport of lipids. Inactivation of lipolytic activity of microorganisms to control infection, inhibition of digestive lipase to control obesity, stimulation of metabolic lipase to reduce hyperlipidemia or procoagulant state, or use of pancreatic lipase supplement in the management of cystic fibrosis are examples of how lipase activity modulation can impact human health.
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The use of traditional medicines of natural origin is being encouraged for the treatment of chronic disorders, as synthetic drugs in such cases may cause unpredictable adverse effects. Berberine, a traditional plant alkaloid, is used in Ayurvedic and Chinese medicine for its antimicrobial and antiprotozoal properties. Interestingly, current clinical research on berberine has revealed its various pharmacological properties and multi-spectrum therapeutic applications. An extensive search in three electronic databases (Unbound Medline, PubMed and ScienceDirect) and internet search engines (Scirus and Google Scholar) were used to identify the clinical studies on berberine, without any time constraints. This review elaborates the recent studies which reveal that with time, the drug has evolved with superior therapeutic activities. In addition, this review will also attract the attention of formulation scientists towards the issues and challenges associated in its drug delivery and the probable approaches that may be explored to help patients reap the maximum benefit of this potentially useful drug. A relatively large number of studies discussed here have revealed the possible areas where this phytochemical constituent can exhibit its therapeutic activities in the treatment of chronic ailments or diseases including diabetes, cancer, depression, hypertension and hypercholesterolemia. The potential of the drug remains to be harvested by designing a suitable formulation that could overcome its inherent low bioavailability.
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Our previous studies illustrated that berberine inhibited adipogenesis in murine-derived 3T3-L1 preadipocytes and human white preadipocytes. In this study, the effects of berberine on the adipogenesis of high-fat diet-induced obesity (FD) or normal diet (ND) mice and possible transcriptional impact are investigated. The results demonstrated that in FD mice, berberine reduced mouse weight gain and food intake and serum glucose, triglyceride, and total cholesterol levels accompanied with a down-regulation of PPARgamma expression and an up-regulation of GATA-3 expression. Berberine had no adverse effects on ND mice. These encouraging findings suggest that berberine has excellent pharmacological potential to prevent obesity.