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IOSR Journal of Pharmacy and Biological Sciences (IOSR-JPBS)
e-ISSN: 2278-3008, p-ISSN:2319-7676. Volume 10, Issue 6 Ver. II (Nov - Dec. 2015), PP 148-154
www.iosrjournals.org
DOI: 10.9790/3008-1062148154 www.iosrjournals.org 148 | Page
Isolation, in vitro antidiabetic, antioxidant activity and molecular
docking studies of pentacyclic triterpenoids from Syzygium
alternifolium(wt.) Walp bark
N.V.L Suvarchala Reddy Va,b*
, M. Akhilac
, C.V.S Subrahmanyamd
, G.
Trimurtulue
,N.M. Raghavendrac,f
a
Depatment of Pharmacology, GokarajuRangaraju College of Pharmacy, Osmania University, Hyderabad,
India
b
Jawaharlal Nehru Technololgical University, Hyderabad, India
c
Depatment of Pharmaceutical Chemistry, GokarajuRangaraju College of Pharmacy, Osmania University,
Hyderabad, India
d
Depatment of Pharmaceutics, GokarajuRangaraju College of Pharmacy, Osmania University, Hyderabad,
India
e
LailaNutraceuticals, Vijayawada, India
f
Department of Applied Pharmacology, FundaçãoOswaldo Cruz, Rio de Janeiro, Brasil
Abstract: Diabetes mellitus type 2 is a metabolic disorder characterized by hyperglycemia including insulin
resistance and relative lack of insulin. Type 2 diabetes is expected to reduce ten-year-shorter life expectancy. In
this research article, we have isolated two terpenoids from the stem bark of Syzygium alternifoliumby column
chromatography.The isolated compounds were identified as friedelin and 3β-friedelinol terpenoids by 1H-NMR,
13C-NMR, and Mass spectroscopy. Isolated compounds were screened for in vitro enzyme inhibition assays
against α-glucosidase & α-amylase; and were found decrease plasma glucose level significantly showing
antidiabetic activity. Isolated compounds also exhibited significant radical scavenging antioxidant activity by
DPPH inhibition assay. The results of in vitro experimentation were confirmed by the molecular docking
studies of the isolated terpenoids on α-glucosidase and α-amylase enzymes.The present researchreveals the
antidiabetic and antioxidant activity of isolated terpenoids from stem bark of Syzygium alternifolium against
diabetes mellitus type 2 disorders.
Keywords:α-glucosidase,α-amylase, DPPH,Syzygiumalternifolium, terpenoids and docking
I. Introduction
Type 2 diabetes is a public health problem that is characterized by tissue resistance to insulin combined
with a relative deficiency in insulin secretion [1,2,]. On the other hand, diminished glucose-induced insulin
secretion with shrinkage in pancreatic cell mass will ultimately lead to postprandial hyperglycemia [3].
Postprandial hyperglycemia is an independent risk factor for cardiovascular disease, stroke and mortality; it
initiates a cascade of pro-atherogenic and pro-thrombotic events [4]. Oxidative stress is also considered to be
the major cause in the development of type-II diabetes. In 2011, about 366 million people suffered with diabetes
globally and this is expected to increase to 552 million by 2030 [5]. One recent study by ICMR-INDIA reported
that about 62.4 million type-II diabetic people are from India. This statistics are expected to increase to 101
million by the year 2030 [6].
Currently, the methods used for treating the various types of diabetes can be classified into oral
hypoglycemic drugs and insulin injection therapy. However, in taking these hypoglycemic drugs, patients still
encounter serious side effects, such as abdominal distention, abdominal pain, diarrhea, gastrointestinal
spasmodic pain, constipation, bowel gurgling, nausea, vomiting, anorexia, fatigue, headache, dizziness, skin
itching and others.An effective inhibition of α-glucosidase enzyme (preventing postprandial hyperglycemia)
and α-amylase (antioxidant activity) is a good strategy for the prevention or treatment of Type 2 diabetes,
hypolipoproteinemia and obesity.
Plant-based products have been determined to function in the regulation pathophysiological signaling
pathways in diabetes and exhibit anti-diabetic activity [7,8]. The hypoglycemic effect of some herbal extracts
have been established in human and animal models of type-II diabetics and some of the conventional drugs
have been derived from the active molecules of these herbs. For example, metformin a less toxic and potent oral
hypoglycemic was developed from plant Galgeaofficinals[9]. Among natural compounds, pentacyclic
triterpenoids (PTs) are a class of pharmacologically active and structurally rich metabolites which are reported
to act on glucose metabolism [10]. PTs may exert their glucose-lowering effect through multi-target pathways,
including α-amylase, α-glucosidase, glycogen phosphorylase, Akt/protein kinase B, protein tyrosine
Isolation, in vitro antidiabetic, antioxidant activity and molecular docking studies of pentacyclic…
DOI: 10.9790/3008-1062148154 www.iosrjournals.org 149 | Page
phosphatase 1B, insulin receptor, the membrane G protein-coupled receptor, glucocorticoid receptor, 11b-
hydroxysteroid dehydrogenase, glyceraldehyde-3-phosphate dehydrogenase and glycerol-3-phosphate
dehydrogenase.
Syzygiumalternifolium(Wt.) Walp. (Myrtaceae) is an endemic aromatic tree, distributed in Assam and
Andhra Pradesh, states of India. Locally it is known as mogi/movi. The plant parts were used traditionally as a
medicine to cure various diseases viz., tender shoots and fruits for dysentery, seeds for diabetes and stem bark
was used to treat gastric ulcers [11]. In the present study, we isolated the active constituents from stem bark of
Syzygium alternifolium. Isolated compounds were identified as friedelin and 3β-friedelinol pentacyclic
terpenoids. Isolated PTs were screened for antioxidant and antidiabetic activities by in vitro DPPH inhibition
assay, α-amylase and α-glucosidaseenzyme inhibition biochemical assays. Furthermore, the mechanism of
action of the isolated PTs was explored by molecular docking analysis.
II. Materials and Methods
2.1 Plant material
The bark of Syzygium alternifolium was collected in the month of November 2011 from Seshachalam
hills of Tirupati, Andhra Pradesh. The botanical identity of the plant was confirmed with the help of botanist,
Department of Botany, S. V. University, Tirupati, Andhra Pradesh, India.
2.2 Extraction
The bark of Syzygium alternifolium was chopped into small pieces, cleaned, powdered, air-dried,
sieved (mesh size = 40) and stored in air tight container at room temperature. The powdered plant material (2
kg) was then extracted sequentially with methanol (MeOH) using soxhlet extractor. The extract (329 gm) was
concentrated to dryness using rotary evaporator. The yield of methanol extract was found to be 16.45%
respectively.
2.3 Isolation
Isolated compounds were identified as terpenoids by positive Liebermann’s Burchardt test and
Salkowski test.The stem bark (2 kg) of the Syzygium alternifolium bark was refluxed with methanol of three
hours. After refluxing the bark with methanol, the total filtrate was concentrated to dryness in vacuum at 40 °C
leaving behind the methanolic extract (329 gm). The bark methanol extract of Syzygium alternifolium was
partitioned with hexane and ethyl acetate. The respective solubles were separated. 310 gm of methanol extract
was partitioned with 1:4 ratio of hexane solvent (1200 ml) by refluxing for two hours. The obtained residue was
further fractionated with ethyl acetate: hexane (1:4), followed by column chromatography. The column was
packed with silica gel (100-120). 5 g of hexane extract was taken and adsorbed with 15 g of silica gel. The
column was filled with 50 g of silica gel. The bed volume for the column is 100 ml. The fraction was further
chromatographed to give compound 1 (170 mg) and compound 2 (100 mg).
2.4 Biological assays
2.4.1In vitro inhibition of intestinal α-glucosidase enzyme
An inhibitory α-glucosidase activity was performed according to the method by Feng[12]. In this
method inhibition of α-glucosidase enzyme was determined spectrophotometrically in a 96-well microtiter plate
which is based on p-nitrophenyl-α-D-glucopyranoside (PNPG) as a substrate. Everything considered the test
samples (Compound 1and 2) 120 μl each was mixed with 20 μl of enzyme solution [0.8 U/ml α-glucosidase in
0.01 M potassium phosphate buffer]. The whole mixture was preincubated at 37 °C prior to initiation of the
reaction by incorporating the substrate. After the preincubation period of 15 min, 20 μl PNPG solutions [5.0
mM PNPG in 0.1 M potassium phosphate buffer (pH 6.8)] was incorporated and then incubated at 37 °C. To
terminate the reaction after 15 min of incubation, 80 μl of 0.2 M Na2CO3 in 0.1 M potassium phosphate buffer
was added to the microtiter plate. The percentage inhibition of α-glucosidase activity was determined by
measuring the absorbance of the resultant mixture was recorded at 405 nm.
2.4.2In vitro inhibition of bacterial α-amylase enzyme
The α-amylase inhibitory activity of isolated compounds from Syzygium alternifoliumbark was
performed according to the method of Wan [13]. Samples acarbose (ALD) were mixed and pre-incubated in 20
mM sodium phosphate buffer (pH 6.7) for 5 min at 37°C. After that the volume of reaction mixture was made
up to 2 ml by incorporation of 1 ml of 2% (w/v) starch dissolved in the buffer, and the whole reaction mixture
was incubated for 5 min at 37°C. After the incubation, 1 ml of di-nitro salicylic acid (DNS) color reagent was
incorporated and kept in boiling water bath precisely for 5 min. Then, the temperature of this mixture was
decreased to room temperature by cooling it on ice and added another 6 ml of deionized water. α-Amylase
inhibitory activity was determined by measuring the absorbance of the mixture at 540 nm.
Isolation, in vitro antidiabetic, antioxidant activity and molecular docking studies of pentacyclic…
DOI: 10.9790/3008-1062148154 www.iosrjournals.org 150 | Page
2.4.3 Spectrophotometric DPPH inhibition assay
Antioxidant capacities of compounds 1, 2 were assessed by DPPH free radical scavenging assay [14].
This method is based on decline of relatively stable radical DPPH to the development of non-radical form in the
presence of hydrogen donating test samples. The test samples depict antioxidant activity by the diminution in
purple colored DPPH to the yellow colored diphenylpicryl hydrazine derivatives. In this method, 2 mL of 0.1
mmol/L solution of DPPH in methanol was blended with 1 mL of each compound 1 and 2. The different
mixtures were incubated at room temperature in a dark room for 30 min. The absorbance was measured at 517
nm and percentage inhibition of radical scavenging was determined.
2.4.4 Molecular docking analysis
Mode of inhibition of compounds 1 (friedelin) and 2 (3β–friedelinol) on the α-glucosidase enzyme was
further assessed by molecular docking analysis using ligand fit of GLIDE software from Schrodinger Inc[15].
The protein α-glucosidase (Pdb Code: 3W37) and α-amylase (Pdb Code: 2QV4) was downloaded from Protein
Data Bank (PDB). These proteins were prepared by filling the missing loops and missing side chains using
protein preparation wizard application of Schrodinger drug design software (Maestro 9.1). The proteins were
further processed by removing nonreactive water molecules, and ligand molecules other than crystal ligand. The
ionized proteins having the lowest penalty were energy minimized using the optimized potential for liquid
simulations 2005 force field incorporated in the Impref tool of Glide programme. Then the grid was generated
in the processed protein by excluding the crystal ligand in the active site using receptor grid generation tool of
Glide programme (van der Waals radius scaling factor was limited to 1.0 with a partial charge cut off of 0.25).
The chemical structures of compounds 1 (friedelin) and 2 (3β–friedelinol) and acarbose was drawn using
ChemBioDraw software. The ligands (Acarbose, Fredeline and Fredelinol) were subjected to Ligprep
simulations to generate energy minimized 3D structures (300 steps) by investigating tautomeric, stereo
chemical, and ionization variations. The ligprepout ligands were docked flexibly in the protein grids using
Glide-extra precision (XP) simulations [16]. Energies of residues within 12 Å of grid were used for simulations.
Poses having coulomb-vdw energy greater than 0.0 kcal/mol and poses having RMS deviation 0.5Å were
discarded. Finally the interactions were scored based on non-bonded interactions.
III. Results and discussion
3.1 Chemistry
The hexane-Et2O (1:1, v/v) extracts of the root barks of stem bark of Syzygium alternifolium was
subjected to multiple chromatographic steps, involving vacuum liquid chromatography, medium-pressure liquid
chromatography and preparative TLC, on silica gel to yield two Friedelane-type triterpenoids. The structures of
the new compounds 1–2(Table 1) were deduced as described below.
3.1.1 Compound 1
The compound 1 was obtained as white crystalline powder with a melting point of 259 °C,Rf(cm) 0.71
(solvent system-hexane: ethyl acetate) (8:2). The molecular formula of compound 1 was shown to be C30H50O
based on the positive ion LC-MS [m/z 427.5 (M+H)+
and 449.4 (M+Na) +
] analysis. The IR spectrum showed
bands at 2972, 2945 (CH3 asymmetric and symmetric stretching); 2895, 2870 (CH2 asymmetric and symmetric
stretching); 1714 (OH stretching), 1454, 1386, 1246 (C=C aromatic skeletal bands). In the 1
H-NMR spectrum
(CDCl3, 400 MHz) quartet signal integrating for one proton at (δ 2.40q) assigned to H-4. The resonances of
eight methyls δ 0.72 (s, 3H, H-24), 0.87 (s, 3H, H-25), 0.87 (d, 3H, J=6.5Hz, H-23), 0.94 (s, 3H, H-30), 1.01 (s,
3H, H-29), 1.003 (s, 3H, H-27), 1.005 (s, 3H, H-26) and 1.18 (s, 3H, H-28). Eleven methylene protons showed
multiplet signals at δ2.2 and 2.30 (H-2); δ1.956 and1.75 (H-1); δ1.70 and 1.58 (H-6:); δ1.58 and1.55 (H-7);
δ1.54 and1.50 (H-11); δ1.45 and1.43 (H-12); δ1.40 and1.39 (H-15); δ δ1.40 and1.39 (H-16); δ1.35 and1.32 (H-
19); δ1.29 and1.28 (H-21); and δ 1.25 and 1.20 (H-22). The remaining three methine protons were seen as
multiplets at δ 1.28 (3H, J = 6.5Hz, H-8, H-10 and H-18). The 13
C-NMR spectrum of compound 1 indicated the
presence of 30 carbon resonances. The DEPT spectrum indicated the presence of carbonyl at δ 213.07 including
seven quaternary carbons, four CH carbons, ten CH2 carbons and eight CH3 carbons. Since the molecular
formula indicative six units of unsaturation, this compound 1 was concluded to be pentacyclic triterpene with a
ketone group. The presence of signals due to one secondary and seven quaternary methyls in the 1
H-NMR
spectrum suggested the Friedelane skeleton. The 1
Hand 13
C-NMR spectral data of compound 1 indicates that it
belongs to was identified as Friedelin. The identification of friedelin was confirmed by comparison of the
reported spectral data with compound [17].
3.1.2 Compound 2
The compound 2 was obtained as white crystalline powder having a melting point of 294°C, Rf(cm)
0.71 (solvent system-hexane: ethyl acetate) (8:2). The molecular formula of compound 2 was shown to be
Isolation, in vitro antidiabetic, antioxidant activity and molecular docking studies of pentacyclic…
DOI: 10.9790/3008-1062148154 www.iosrjournals.org 151 | Page
C30H52O based on the positive ion LC-MS [m/z 428.4(M)+
and 429.3 (M+H)+
] analysis. The IR spectrum
showed bands at 3473 (OH stretching),3003 (aromatic CH stretching), 2978, 2943 (CH3asymmetric and
symmetric stretch), 2872, 2837 (CH2 asymmetric and symmetric stretch) and 1477, 1384 (aromatic C=C skeletal
bands). The 1
H NMR spectrum (CDCl3, 400 MHz) gave a peak at δ 3.73 which corresponds to H-3. In the
HMBC spectrum C-1 showed correlations with H-3 and H-10. H-3 resonated at δ 3.73, ddJ = 10.8, 4.6 Hz, the
HMBC correlations and this chemical shift value allowed us to place the hydroxyl functionality at this position
(C-3). The resonances of eight methyls δ 0.85 (s, 3H, H-25), 0.93 (s, 3H, H-24), 1.06 (s, 6H, H-29, H-30), 0.99
(d, 3H, H-23), 1.00 (s, 3H, H-27), 1.01 (s, 3H, H-26), 1.16 (s, 3H, H-28). Eleven methylene protons showed
multiplet signals at δ1.90 and 1.75 (H-2); δ1.85 and1.70 (H-1); δ1.53 and 1.50 (H-6:); δ1.51 and1.49 (H-7);
δ1.47 and1.39 (H-11); δ1.45 and1.43 (H-12); δ1.43 and1.39 (H-15); δ1.42 and1.38 (H-16); δ1.36 and1.34 (H-
19); δ1.35 and1.33 (H-21); and δ 1.24 and 1.19 (H-22). Out of remaining four methine protons, methine proton
showed multiplet at δ 1.55 (m, J= 6.0Hz, H-4), while others were seen at 1.25-1.32 (3H, J = 6.8Hz, H-8, H-10
and H-18).H-10 resonated at δ 1.25, ddJ=6.8 Hz the multiplicity, dd was due to a coupling of this proton with
H-1 proton. C-3 (δ- 72.20) showed correlations with H-2 and H-23. These correlations and the chemical shift
value supported to phase the hydroxyl functionality at C-3 and methyl a group (C-23) at C-4. Further, in the 1D
proton NMR spectrum, the methyl group at C-23 split in to a doublets, which was due to the coupling of these
methyl protons with H-4 and fits the characteristic of this methyl group (C-23) at C-4. C-4 showed correlation
with H-2, H-23 and H-24 and the quaternary carbon C-3 showed correlations with H-1, H-3, H-6 and H-24.
These correlations supported the attachment of C-23 methyl group at C-4 and C-24 methyl group at C-5. The
methyl protons at H-26 showed correlations with C-12, C-13, C-14 and C-18. These correlations allowed us to
place the C-26 methyl group at C-13 and C-27 methyl a group at C-14 respectively. The methyl protons at H-28
showed correlations with C-16, C-17, C-18 and C-22. These correlations allowed us to place C-28 methyl group
at C-17. Finally, C-29 showed correlations with H-19, H-21 and H-30; H-30 showed correlations with H-19, H-
21 and H-29. C-20 showed correlations with H-19, H-21, H-29 and H-30; C-21 showed correlations with H-
22,H-29 and H-30 all these correlations supported the attachment of both C-29 and C-30 methyl groups to the
same carbon at C-20 and the compound is an oleane type triterpenoid. The β orientation of the hydroxyl group
at C-3 was confirmed by comparing the proton coupling between H-3 and H-4 and H-2 confirms the
stereochemistry.
The 13
C-NMR spectrum of 2 indicated the presence of 30 carbon resonances. The DEPT spectrum
indicated the presence of six quaternary carbons, five CH carbons, eleven CH2 carbons and eight CH3 carbons;
one of the carbons resonated at δ72.8 and indicative of presence of hydroxyl group.These data of compound 2
had good agreements with the reported data of 3β–friedelinol[18].
3.1.3 Biological evaluation
Pentacyclic triterpenoids (PTs) have been reported as naturally occurring active constituents having
regulatory action on glucose metabolism by many biochemical mechanisms including inhibition of α-
glucosidase and α-amylase. α-glucosidase is membrane bound enzyme integrated in the epithelium of the small
intestine and is implicated for the last step of carbohydrate hydrolysis to create absorbable monosaccharide.
Starch is the main dietary carbohydrate cause of glucose and diabetic complications depends upon the rate and
extent of digestion of glucose from starch by the pancreatic α-amylase and intestinal α-glucosidase [19].
Disaccharides and oligosaccharides are broken down by the pancreatic α-amylase, followed by
intestinal α-glucosidase enzyme which catabolizes disaccharides into glucose [20].The isolated
terpenoidscompounds were evaluated for the inhibitory activity against α-glucosidase and α-amylase protein.
Table 2 shows the inhibitory activity of terpenoidsagainst α-glucosidase and α-amylase in the percentage
inhibition at 10-50µM concentration. Compound 2 (3β–friedelinol) inhibited the enzymes α-glucosidase and α-
amylase with 15.57 and 36.35 percentage respectively and compound 1 (fredelin) inhibited the enzymes α-
glucosidase and α-amylase with 10.4 and 23.04 percentage respectively. These terpenoids have shown better
antidiabetic activity than marketed α-glucosidase inhibitor acarbose, which showed the inhibition of 9.46
percentages against both α-glucosidase and α-amylase. These results were in agreement to the in vitro α-
glucosidase inhibition of terpenoids as per reported literature [10,21]. Antioxidants have been known to exert
the protective effects against the oxidative damage and are associated with reduced risk of chronic diseases such
as inflammation, diabetes, carcinogenesis and atherosclerosis [21,22,23]. Isolated PTs when investigated for
their efficacy in scavenging free radicals through DPPH inhibition assay; Compound 2 (3β–friedelinol) was
found to have better antioxidant activity, followed by compound 1 (fredeline) (Table 3).
3.1.4 Molecular Modeling
In this research work, we also sought to give in silico evidence for the binding interaction of isolated
compounds 1 (Fredeline) and 2 (3β–friedelinol) against α-glucosidase and α-amylase enzymes. The interactions
were scored based on non-bonded interactions such as lipophilic pair term, hydrophobic enclosure reward,
Isolation, in vitro antidiabetic, antioxidant activity and molecular docking studies of pentacyclic…
DOI: 10.9790/3008-1062148154 www.iosrjournals.org 152 | Page
hydrogen bonding, and electrostatic rewards (Table 4 and Table 5). As expected, compounds 1 and 2 were
shown to have efficient antagonism of α-glucosidase and α-amylase enzymes. Acarbose is a tetrasaccharide
possessing multiple free hydroxyl groups capable of forming numerous polar interactions with the active site of
protein.While, compounds 1 (fredeline) and 2 (3β–friedelinol) being non polar in nature, exhibited enhanced
hydrophobic interactions to inhibit α-glucosidase and α-amylase enzymes (Fig 1 and Fig 2). With α-glucosidase
enzyme (Pdbcode: 3W37), compounds 1 (fredeline) and 2 (3β–friedelinol) possess hydrophobic interactions
with Phe476, Trp432, Ile 358, Trp329, Phe601, Ala602 aminoacids. The hydrophobic interaction in acarbose
were comparitively less (Phe 476, Trp432, Met470, Phe601) than polar hydrogen bonding interactions(His626,
Asp357(2), Asp568, Arg552, Asp232). With α-amylase protein (Pdb code: 2QV4), the polar interaction are due
to the hydroxyl group of 3β–friedelinol with Hie305 amino acid. The non-polar interactions of compounds 1
and 2 were found to be with Glu60, Trp58, Leu165, Val107, Ala106 aminoacids. Acarbose also possess similar
interactions with additional hydrogen bonding with various amino acids (Asp300, Arg195, Glu233, Thr163, and
Gln63).
IV. Conclusion
In summary, we report the isolation and characterization of Friedelin and 3β–Friedelinol pentacyclic
terpenoids from the stem bark of Syzygium alternifolium. To our knowledge nobody has reported the
identification of these two PTs from Syzygium alternifolium. The isolated compounds 1 (fredeline) and 2 (3β–
friedelinol) PTs showed a showed significant inhibition of α-glucosidase, α-amylase and DPPH indicating the
reduction in high blood glucose level as well as the reduction in the formation of ROS (Reactive Oxygen
Species). Inhibition of these enzymes by compounds 1 (fredeline) and 2 (3β–friedelinol) was further supported
by the molecular docking analysis. Therefore, these properties would be of therapeutic benefit in insulin
resistant states. Future research will be carried out in this direction.
Acknowledgements:
We are grateful to the management of GokarajuRangaraju College of Pharmacy, Hyderabad and
LailaNutraceuticals, Vijayawada for their support and cooperation.
References
[1]. Bazzano, L. Serdula, M.L.S. Prevention of type 2 diabetes by diet and lifestyle modification. J. Am. Coll. Nutr. 24, 2005, 310-319.
[2]. Ahmed, D. Sharma, M. Mukerjee, A. Ramteke, P.W. Kumar, V. Improved glycemic control, pancreas protective and
hepatoprotective effect by traditional poly-herbal formulation “QursTabasheer” in streptozotocin induced diabetic rats. BMC
Complementary and Alternative Medicine, 2013, doi: 10.1186/1472-6882-13-10.
[3]. Kumar, V. Ahmed, D. Gupta, P.S. Anwar, F. Mujeeb, M. Anti-diabetic, anti-oxidant and antihyperlipidemic activities of
Melastomamalabathricum Linn. leaves in streptozotocin induced diabetic rats. BMC Complementary and Alternative Medicine
13,2013, 222.
[4]. Ferrannini, E. Gastaldelli, A. Miyazaki, Y. Matsuda, M. Mari, A. DeFronzo, R. Beta-cell function in subjects spanning the range
from normal glucose tolerance to overt diabetes: a new analysis. The Journal of Clinical Endocrinology and Metabolism, 90,2005,
493-500.
[5]. Whiting, D.R. Guariguata, L. Weil, C. Shaw, J. IDF diabetes atlas: Global estimates of the prevalence of diabetes for 2011 and
2030. Diabetes Research and Clinical Practice, 94,2011, 311-321.
[6]. Anjana, R.M. Pradeepa, R. Deepa, M. Datta, M. Sudha, V. Unnikrishnan, R. Prevalence of diabetes and prediabetes (impaired
fasting glucose and/or impaired glucose tolerance) in urban and rural India: Phase I results of the Indian Council of Medical
Research-India diabetes (ICMR-INDIAB) study. Diabetologia, 54,2011, 3022-3027.
[7]. Hung, H.Y. Qian, K. Morris-Natschke, S.L. Hsu, C.S. Lee, K.H.Recent discovery of plant-derived anti-diabetic natural
products.Natural Products Reports, 29, 2012, 580-606.
[8]. Ahmed, D. Kumar, V. Sharma, M. Verma, A. Target guided isolation, in-vitro antidiabetic, antioxidant activity and molecular
docking studies of some flavonoids from AlbizzialebbeckBenth. Bark. BMC Complementary and Alternative Medicine, 14,2014,
155.
9. Bailey, C. Day, C. Metformin: Its botanical background. Practical Diabetes International, 21,2004, 115-117.
[9]. Ardiles, A.E. Rodriguez, A.G. Nunez, M.J. Perestelo, N.R. Studies of naturally occurring friedelanetriterpenoids as insulin
sensitizers in the treatment of type 2 diabetes mellitus. Phytochemistry, 84,2012, 116-124.
[10]. Ramesh, B. K. Abdul Nabi, S. Swapna, S. Apparao, C.Cinnamic acid as one of the antidiabetic active principle(s) from the seeds of
Syzygium alternifolium. Food and Chemical and Toxicology, 50,2012, 1425-1431.
[11]. Feng, J. Yang, X.W. Wang, R.F. Bio-assay guided isolation and identification of α-glucosidase inhibitors from the leaves of
Aquilariasinensis. Phytochemistry,72,2011, 242-247.
[12]. Wan, L.S. Chen, C.P. Xiao, Z.Q. Wang, Y.L. Min, Q.X. Yue, Y. Chen, J. In vitro and in vivo anti-diabetic activity of
Swertiakouitchensis extract. Journal of Ethnopharmacology, 147, 2013, 622-630.
[13]. Zhang, Y. Li, Q. Xing, H. Xuefeng, L. Zhao, L. Kankan, Q. Bi, K. Evaluation of antioxidant activity of ten compounds in different
tea samples by means of an on-line HPLC–DPPH assay. Food Research International, 53,2013, 847-856.
[14]. Raghavendra, N.M. Jyothsna, A. VenkateswaraRao, A. Subrahmanyam, C.V.S. Synthesis, pharmacological evaluation and docking
studies of N-(benzo[d]thiazol-2-yl)-2-(piperazin-1- yl)acetamide analogs as COX-2 inhibitors. Bioorganic and Medicinal Chemistry
Letters, 22,2012, 820-823.
[15]. Raghavendra, N.M. Ramakrishna, K.Sirisha, V.Divya, P.VenkateswaraRao,A. Computer aided discovery of potential anti-
inflammatory (s)-naproxen analogs as COX-2 inhibitors. Medicinal Chemistry,9, 2013, 553-559.
Isolation, in vitro antidiabetic, antioxidant activity and molecular docking studies of pentacyclic…
DOI: 10.9790/3008-1062148154 www.iosrjournals.org 153 | Page
[16]. Hishashi, M. Toshiyuki, M. Yashiro, K.Johji, Y. Masayuki, Y. Antidiabetic principles of natural medicines. IV. Aldose reductase
and α-glucosidase inhibitors from the roots of Salaciaoblonga Wall (Celastraceae): Structure of a new friedelane-type triterpene,
kotalagenin 16- acetate. Chemical and Pharmaceutical Bulletin, 47,1999, 1725-1729.
[17]. David Dako, C. Hanh Trinh, V.D. Wansi, J.D.New friedelane triterpenoids with antimicrobial activity from the stems of
Drypetespaxii.Chemical and Pharmaceutical Bulletin, 57, 2009, 1119-1122.
[18]. Dhital, S. Lin, AH-M. Hamaker, B.R. Gidley, M.J. Muniandy, A. Mammalian mucosal α-glucosidases coordinate with α-amylase
in the initial starch hydrolysis stage to have a role in starch digestion beyond glucogenesis. PLoS One, 8, 2013, e62546.
[19]. Kwon, Y.I. Apostolidis, E. Kim, Y.C. Shetty, K. Health benefits of traditional corn, beans and pumpkin: In vitro studies for
hyperglycemia and hypertension management. Journal of Medicinal Food, 10,2007, 266-275.
[20]. Tadera, K. Minami, Y. Takamatsu, K. Matsuoka, T. Inhibition of α-glucosidase and α-amylase by flavanoids. Journal of Nutritional
Science and Vitaminology, 52,2006, 149-153.
[21]. Liu, R.H. Adom, K.K. Antioxidant activity of grains. Journal of Agricultural and Food Chemistry, 50,2002, 6182-6187.
[22]. Maria, S.M.R. Fabiano, A.N.F. Ricardo, E.A. Debrito, E.S. Free radical scavenging behavior of some north-east Brazilian fruits in
a DPPH system. Food Chemistry, 114,2009, 693-695.
[23]. Anderson, R.A. Evans, M.L. Ellis, G.R. Graham, J. Morris, K. Jackson, S.K. Lewis, M.J. Rees, A. Frenneaux, M.P.The relationship
between post-prandial lipaemia, endothelial function and oxidative stress in healthy individuals and patients with type 2
diabetes.Atherosclerosis, 154, 2001, 475-483.
Table 1: Structural details of isolated compounds from the bark of Syzygiumalternifolium
Compound code Chemical structure IUPAC name Common names
Compound 1
2
3
4
5
10
1
6
7
8
9 14
13
12
11
15
16
17
18
22
21
20
19
O
23
24
25
27
28
26
30 29
(4R,4aS,6bR,8aR,12bS,14aS)-
octadecahydro-
4,4a,6b,8a,11,11,12b,14a-
octamethylpicen-3(4H,6bH,14bH)-
one
Friedelin
Compound 2 2
3
4
5
10
1
6
7
8
9 14
13
12
11
15
16
17
18
22
21
20
19
HO
23
24
25
27
28
26
30 29
(3S,4R,4aS,6bR,8aR,12bS,14aS)-
docosahydro-
4,4a,6b,8a,11,11,12b,14a-
octamethylpicen-3-ol
3β–Friedelinol
Table 2: In vitro enzyme inhibition activity against α- glucosidase and α-amylase
α-Glucosidase α-Amylase
Compound IC50 value ± SEM IC50 value ± SEM
Compound 1 10.40 ± 0.12** 23.04 ± 0.34*
Compound 2 15.57 ± 0.23** 36.35 ± 0.21**
Acarbose 09.46 ± 0.02 09.46 ± 0.02
SEM = Standard Error Mean; **p < 0.001, ***p < 0.0001
Table 3: Inhibition of radical scavenging activity by DPPH assay
Samples IC50 value ± SEM
Compound 1 20.5*** ± 0.39
Compound 2 30.5*** ± 0.22
Ascorbic acid 17.3 ± 1.10
SEM = Standard Error Mean; ***p < 0.0001
Table 4: Scoring Data (kcal/mol) of Glide-XP Docking of Schrodinger (Maestro 9.1) against α-amylase
enzyme(PDB Code: 2QV4)
Ligand Docking Scorea Lipophilic
EvdWb H-Bondc
Electrod
RotPenale
Acarbose -12.37 -3.87 -4.80 -1.95 0.10
Compound 2 -6.05 -3.27 -0.68 -0.13 0.00
Compound 1 -5.43 -2.93 0.00 -0.01 0.00
a
An estimate of ligand- α-amylase binding energy; b
Lipophilic van der Walls interactions of ligand- α-amylase
complex; c
Hydrogen-bonding term; d
Electrostatic interactions of ligand- α-amylase complex; e
Rotational
penalty causing the decreased ligand-protein interactions.
Isolation, in vitro antidiabetic, antioxidant activity and molecular docking studies of pentacyclic…
DOI: 10.9790/3008-1062148154 www.iosrjournals.org 154 | Page
Table 5: Scoring Data (kcal/mol) of Glide-XP Docking of Schrodinger (Maestro 9.1) against α-
glucosidase enzyme (PDB Code: 3W37)
Ligand
Docking
Scorea Lipophilic EvdWb
H-Bondc
Electrod
RotPenale
Acarbose -10.5 -1.94 -4.87 -2.00 0.00
Compound 1 -3.28 -1.17 0.00 -0.12 0.00
Compound 2 -3.57 -1.36 -0.70 -0.11 0.00
a
An estimate of ligand- α-amylase binding energy; b
Lipophilic van der Walls interactions of ligand- α-amylase
complex; c
Hydrogen-bonding term; d
Electrostatic interactions of ligand- α-amylase complex; e
Rotational
penalty causing the decreased ligand-protein interactions.
Fig. 1 Superimposition of compound 1, compound 2 and acarbose (line forms) in the binding site of α-
amylase (PDB Code: 2QV4).
Fig. 2 Overlay view of compounds 1, 2 and acarbose (stick forms) in α-glucosidase binding pocket
surrounded by non-polar amino acids (space fill form) (PDB Code: 3W37).

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Isolation, in vitro antidiabetic, antioxidant activity and molecular docking studies of pentacyclic triterpenoids from Syzygium alternifolium(wt.) Walp bark

  • 1. IOSR Journal of Pharmacy and Biological Sciences (IOSR-JPBS) e-ISSN: 2278-3008, p-ISSN:2319-7676. Volume 10, Issue 6 Ver. II (Nov - Dec. 2015), PP 148-154 www.iosrjournals.org DOI: 10.9790/3008-1062148154 www.iosrjournals.org 148 | Page Isolation, in vitro antidiabetic, antioxidant activity and molecular docking studies of pentacyclic triterpenoids from Syzygium alternifolium(wt.) Walp bark N.V.L Suvarchala Reddy Va,b* , M. Akhilac , C.V.S Subrahmanyamd , G. Trimurtulue ,N.M. Raghavendrac,f a Depatment of Pharmacology, GokarajuRangaraju College of Pharmacy, Osmania University, Hyderabad, India b Jawaharlal Nehru Technololgical University, Hyderabad, India c Depatment of Pharmaceutical Chemistry, GokarajuRangaraju College of Pharmacy, Osmania University, Hyderabad, India d Depatment of Pharmaceutics, GokarajuRangaraju College of Pharmacy, Osmania University, Hyderabad, India e LailaNutraceuticals, Vijayawada, India f Department of Applied Pharmacology, FundaçãoOswaldo Cruz, Rio de Janeiro, Brasil Abstract: Diabetes mellitus type 2 is a metabolic disorder characterized by hyperglycemia including insulin resistance and relative lack of insulin. Type 2 diabetes is expected to reduce ten-year-shorter life expectancy. In this research article, we have isolated two terpenoids from the stem bark of Syzygium alternifoliumby column chromatography.The isolated compounds were identified as friedelin and 3β-friedelinol terpenoids by 1H-NMR, 13C-NMR, and Mass spectroscopy. Isolated compounds were screened for in vitro enzyme inhibition assays against α-glucosidase & α-amylase; and were found decrease plasma glucose level significantly showing antidiabetic activity. Isolated compounds also exhibited significant radical scavenging antioxidant activity by DPPH inhibition assay. The results of in vitro experimentation were confirmed by the molecular docking studies of the isolated terpenoids on α-glucosidase and α-amylase enzymes.The present researchreveals the antidiabetic and antioxidant activity of isolated terpenoids from stem bark of Syzygium alternifolium against diabetes mellitus type 2 disorders. Keywords:α-glucosidase,α-amylase, DPPH,Syzygiumalternifolium, terpenoids and docking I. Introduction Type 2 diabetes is a public health problem that is characterized by tissue resistance to insulin combined with a relative deficiency in insulin secretion [1,2,]. On the other hand, diminished glucose-induced insulin secretion with shrinkage in pancreatic cell mass will ultimately lead to postprandial hyperglycemia [3]. Postprandial hyperglycemia is an independent risk factor for cardiovascular disease, stroke and mortality; it initiates a cascade of pro-atherogenic and pro-thrombotic events [4]. Oxidative stress is also considered to be the major cause in the development of type-II diabetes. In 2011, about 366 million people suffered with diabetes globally and this is expected to increase to 552 million by 2030 [5]. One recent study by ICMR-INDIA reported that about 62.4 million type-II diabetic people are from India. This statistics are expected to increase to 101 million by the year 2030 [6]. Currently, the methods used for treating the various types of diabetes can be classified into oral hypoglycemic drugs and insulin injection therapy. However, in taking these hypoglycemic drugs, patients still encounter serious side effects, such as abdominal distention, abdominal pain, diarrhea, gastrointestinal spasmodic pain, constipation, bowel gurgling, nausea, vomiting, anorexia, fatigue, headache, dizziness, skin itching and others.An effective inhibition of α-glucosidase enzyme (preventing postprandial hyperglycemia) and α-amylase (antioxidant activity) is a good strategy for the prevention or treatment of Type 2 diabetes, hypolipoproteinemia and obesity. Plant-based products have been determined to function in the regulation pathophysiological signaling pathways in diabetes and exhibit anti-diabetic activity [7,8]. The hypoglycemic effect of some herbal extracts have been established in human and animal models of type-II diabetics and some of the conventional drugs have been derived from the active molecules of these herbs. For example, metformin a less toxic and potent oral hypoglycemic was developed from plant Galgeaofficinals[9]. Among natural compounds, pentacyclic triterpenoids (PTs) are a class of pharmacologically active and structurally rich metabolites which are reported to act on glucose metabolism [10]. PTs may exert their glucose-lowering effect through multi-target pathways, including α-amylase, α-glucosidase, glycogen phosphorylase, Akt/protein kinase B, protein tyrosine
  • 2. Isolation, in vitro antidiabetic, antioxidant activity and molecular docking studies of pentacyclic… DOI: 10.9790/3008-1062148154 www.iosrjournals.org 149 | Page phosphatase 1B, insulin receptor, the membrane G protein-coupled receptor, glucocorticoid receptor, 11b- hydroxysteroid dehydrogenase, glyceraldehyde-3-phosphate dehydrogenase and glycerol-3-phosphate dehydrogenase. Syzygiumalternifolium(Wt.) Walp. (Myrtaceae) is an endemic aromatic tree, distributed in Assam and Andhra Pradesh, states of India. Locally it is known as mogi/movi. The plant parts were used traditionally as a medicine to cure various diseases viz., tender shoots and fruits for dysentery, seeds for diabetes and stem bark was used to treat gastric ulcers [11]. In the present study, we isolated the active constituents from stem bark of Syzygium alternifolium. Isolated compounds were identified as friedelin and 3β-friedelinol pentacyclic terpenoids. Isolated PTs were screened for antioxidant and antidiabetic activities by in vitro DPPH inhibition assay, α-amylase and α-glucosidaseenzyme inhibition biochemical assays. Furthermore, the mechanism of action of the isolated PTs was explored by molecular docking analysis. II. Materials and Methods 2.1 Plant material The bark of Syzygium alternifolium was collected in the month of November 2011 from Seshachalam hills of Tirupati, Andhra Pradesh. The botanical identity of the plant was confirmed with the help of botanist, Department of Botany, S. V. University, Tirupati, Andhra Pradesh, India. 2.2 Extraction The bark of Syzygium alternifolium was chopped into small pieces, cleaned, powdered, air-dried, sieved (mesh size = 40) and stored in air tight container at room temperature. The powdered plant material (2 kg) was then extracted sequentially with methanol (MeOH) using soxhlet extractor. The extract (329 gm) was concentrated to dryness using rotary evaporator. The yield of methanol extract was found to be 16.45% respectively. 2.3 Isolation Isolated compounds were identified as terpenoids by positive Liebermann’s Burchardt test and Salkowski test.The stem bark (2 kg) of the Syzygium alternifolium bark was refluxed with methanol of three hours. After refluxing the bark with methanol, the total filtrate was concentrated to dryness in vacuum at 40 °C leaving behind the methanolic extract (329 gm). The bark methanol extract of Syzygium alternifolium was partitioned with hexane and ethyl acetate. The respective solubles were separated. 310 gm of methanol extract was partitioned with 1:4 ratio of hexane solvent (1200 ml) by refluxing for two hours. The obtained residue was further fractionated with ethyl acetate: hexane (1:4), followed by column chromatography. The column was packed with silica gel (100-120). 5 g of hexane extract was taken and adsorbed with 15 g of silica gel. The column was filled with 50 g of silica gel. The bed volume for the column is 100 ml. The fraction was further chromatographed to give compound 1 (170 mg) and compound 2 (100 mg). 2.4 Biological assays 2.4.1In vitro inhibition of intestinal α-glucosidase enzyme An inhibitory α-glucosidase activity was performed according to the method by Feng[12]. In this method inhibition of α-glucosidase enzyme was determined spectrophotometrically in a 96-well microtiter plate which is based on p-nitrophenyl-α-D-glucopyranoside (PNPG) as a substrate. Everything considered the test samples (Compound 1and 2) 120 μl each was mixed with 20 μl of enzyme solution [0.8 U/ml α-glucosidase in 0.01 M potassium phosphate buffer]. The whole mixture was preincubated at 37 °C prior to initiation of the reaction by incorporating the substrate. After the preincubation period of 15 min, 20 μl PNPG solutions [5.0 mM PNPG in 0.1 M potassium phosphate buffer (pH 6.8)] was incorporated and then incubated at 37 °C. To terminate the reaction after 15 min of incubation, 80 μl of 0.2 M Na2CO3 in 0.1 M potassium phosphate buffer was added to the microtiter plate. The percentage inhibition of α-glucosidase activity was determined by measuring the absorbance of the resultant mixture was recorded at 405 nm. 2.4.2In vitro inhibition of bacterial α-amylase enzyme The α-amylase inhibitory activity of isolated compounds from Syzygium alternifoliumbark was performed according to the method of Wan [13]. Samples acarbose (ALD) were mixed and pre-incubated in 20 mM sodium phosphate buffer (pH 6.7) for 5 min at 37°C. After that the volume of reaction mixture was made up to 2 ml by incorporation of 1 ml of 2% (w/v) starch dissolved in the buffer, and the whole reaction mixture was incubated for 5 min at 37°C. After the incubation, 1 ml of di-nitro salicylic acid (DNS) color reagent was incorporated and kept in boiling water bath precisely for 5 min. Then, the temperature of this mixture was decreased to room temperature by cooling it on ice and added another 6 ml of deionized water. α-Amylase inhibitory activity was determined by measuring the absorbance of the mixture at 540 nm.
  • 3. Isolation, in vitro antidiabetic, antioxidant activity and molecular docking studies of pentacyclic… DOI: 10.9790/3008-1062148154 www.iosrjournals.org 150 | Page 2.4.3 Spectrophotometric DPPH inhibition assay Antioxidant capacities of compounds 1, 2 were assessed by DPPH free radical scavenging assay [14]. This method is based on decline of relatively stable radical DPPH to the development of non-radical form in the presence of hydrogen donating test samples. The test samples depict antioxidant activity by the diminution in purple colored DPPH to the yellow colored diphenylpicryl hydrazine derivatives. In this method, 2 mL of 0.1 mmol/L solution of DPPH in methanol was blended with 1 mL of each compound 1 and 2. The different mixtures were incubated at room temperature in a dark room for 30 min. The absorbance was measured at 517 nm and percentage inhibition of radical scavenging was determined. 2.4.4 Molecular docking analysis Mode of inhibition of compounds 1 (friedelin) and 2 (3β–friedelinol) on the α-glucosidase enzyme was further assessed by molecular docking analysis using ligand fit of GLIDE software from Schrodinger Inc[15]. The protein α-glucosidase (Pdb Code: 3W37) and α-amylase (Pdb Code: 2QV4) was downloaded from Protein Data Bank (PDB). These proteins were prepared by filling the missing loops and missing side chains using protein preparation wizard application of Schrodinger drug design software (Maestro 9.1). The proteins were further processed by removing nonreactive water molecules, and ligand molecules other than crystal ligand. The ionized proteins having the lowest penalty were energy minimized using the optimized potential for liquid simulations 2005 force field incorporated in the Impref tool of Glide programme. Then the grid was generated in the processed protein by excluding the crystal ligand in the active site using receptor grid generation tool of Glide programme (van der Waals radius scaling factor was limited to 1.0 with a partial charge cut off of 0.25). The chemical structures of compounds 1 (friedelin) and 2 (3β–friedelinol) and acarbose was drawn using ChemBioDraw software. The ligands (Acarbose, Fredeline and Fredelinol) were subjected to Ligprep simulations to generate energy minimized 3D structures (300 steps) by investigating tautomeric, stereo chemical, and ionization variations. The ligprepout ligands were docked flexibly in the protein grids using Glide-extra precision (XP) simulations [16]. Energies of residues within 12 Å of grid were used for simulations. Poses having coulomb-vdw energy greater than 0.0 kcal/mol and poses having RMS deviation 0.5Å were discarded. Finally the interactions were scored based on non-bonded interactions. III. Results and discussion 3.1 Chemistry The hexane-Et2O (1:1, v/v) extracts of the root barks of stem bark of Syzygium alternifolium was subjected to multiple chromatographic steps, involving vacuum liquid chromatography, medium-pressure liquid chromatography and preparative TLC, on silica gel to yield two Friedelane-type triterpenoids. The structures of the new compounds 1–2(Table 1) were deduced as described below. 3.1.1 Compound 1 The compound 1 was obtained as white crystalline powder with a melting point of 259 °C,Rf(cm) 0.71 (solvent system-hexane: ethyl acetate) (8:2). The molecular formula of compound 1 was shown to be C30H50O based on the positive ion LC-MS [m/z 427.5 (M+H)+ and 449.4 (M+Na) + ] analysis. The IR spectrum showed bands at 2972, 2945 (CH3 asymmetric and symmetric stretching); 2895, 2870 (CH2 asymmetric and symmetric stretching); 1714 (OH stretching), 1454, 1386, 1246 (C=C aromatic skeletal bands). In the 1 H-NMR spectrum (CDCl3, 400 MHz) quartet signal integrating for one proton at (δ 2.40q) assigned to H-4. The resonances of eight methyls δ 0.72 (s, 3H, H-24), 0.87 (s, 3H, H-25), 0.87 (d, 3H, J=6.5Hz, H-23), 0.94 (s, 3H, H-30), 1.01 (s, 3H, H-29), 1.003 (s, 3H, H-27), 1.005 (s, 3H, H-26) and 1.18 (s, 3H, H-28). Eleven methylene protons showed multiplet signals at δ2.2 and 2.30 (H-2); δ1.956 and1.75 (H-1); δ1.70 and 1.58 (H-6:); δ1.58 and1.55 (H-7); δ1.54 and1.50 (H-11); δ1.45 and1.43 (H-12); δ1.40 and1.39 (H-15); δ δ1.40 and1.39 (H-16); δ1.35 and1.32 (H- 19); δ1.29 and1.28 (H-21); and δ 1.25 and 1.20 (H-22). The remaining three methine protons were seen as multiplets at δ 1.28 (3H, J = 6.5Hz, H-8, H-10 and H-18). The 13 C-NMR spectrum of compound 1 indicated the presence of 30 carbon resonances. The DEPT spectrum indicated the presence of carbonyl at δ 213.07 including seven quaternary carbons, four CH carbons, ten CH2 carbons and eight CH3 carbons. Since the molecular formula indicative six units of unsaturation, this compound 1 was concluded to be pentacyclic triterpene with a ketone group. The presence of signals due to one secondary and seven quaternary methyls in the 1 H-NMR spectrum suggested the Friedelane skeleton. The 1 Hand 13 C-NMR spectral data of compound 1 indicates that it belongs to was identified as Friedelin. The identification of friedelin was confirmed by comparison of the reported spectral data with compound [17]. 3.1.2 Compound 2 The compound 2 was obtained as white crystalline powder having a melting point of 294°C, Rf(cm) 0.71 (solvent system-hexane: ethyl acetate) (8:2). The molecular formula of compound 2 was shown to be
  • 4. Isolation, in vitro antidiabetic, antioxidant activity and molecular docking studies of pentacyclic… DOI: 10.9790/3008-1062148154 www.iosrjournals.org 151 | Page C30H52O based on the positive ion LC-MS [m/z 428.4(M)+ and 429.3 (M+H)+ ] analysis. The IR spectrum showed bands at 3473 (OH stretching),3003 (aromatic CH stretching), 2978, 2943 (CH3asymmetric and symmetric stretch), 2872, 2837 (CH2 asymmetric and symmetric stretch) and 1477, 1384 (aromatic C=C skeletal bands). The 1 H NMR spectrum (CDCl3, 400 MHz) gave a peak at δ 3.73 which corresponds to H-3. In the HMBC spectrum C-1 showed correlations with H-3 and H-10. H-3 resonated at δ 3.73, ddJ = 10.8, 4.6 Hz, the HMBC correlations and this chemical shift value allowed us to place the hydroxyl functionality at this position (C-3). The resonances of eight methyls δ 0.85 (s, 3H, H-25), 0.93 (s, 3H, H-24), 1.06 (s, 6H, H-29, H-30), 0.99 (d, 3H, H-23), 1.00 (s, 3H, H-27), 1.01 (s, 3H, H-26), 1.16 (s, 3H, H-28). Eleven methylene protons showed multiplet signals at δ1.90 and 1.75 (H-2); δ1.85 and1.70 (H-1); δ1.53 and 1.50 (H-6:); δ1.51 and1.49 (H-7); δ1.47 and1.39 (H-11); δ1.45 and1.43 (H-12); δ1.43 and1.39 (H-15); δ1.42 and1.38 (H-16); δ1.36 and1.34 (H- 19); δ1.35 and1.33 (H-21); and δ 1.24 and 1.19 (H-22). Out of remaining four methine protons, methine proton showed multiplet at δ 1.55 (m, J= 6.0Hz, H-4), while others were seen at 1.25-1.32 (3H, J = 6.8Hz, H-8, H-10 and H-18).H-10 resonated at δ 1.25, ddJ=6.8 Hz the multiplicity, dd was due to a coupling of this proton with H-1 proton. C-3 (δ- 72.20) showed correlations with H-2 and H-23. These correlations and the chemical shift value supported to phase the hydroxyl functionality at C-3 and methyl a group (C-23) at C-4. Further, in the 1D proton NMR spectrum, the methyl group at C-23 split in to a doublets, which was due to the coupling of these methyl protons with H-4 and fits the characteristic of this methyl group (C-23) at C-4. C-4 showed correlation with H-2, H-23 and H-24 and the quaternary carbon C-3 showed correlations with H-1, H-3, H-6 and H-24. These correlations supported the attachment of C-23 methyl group at C-4 and C-24 methyl group at C-5. The methyl protons at H-26 showed correlations with C-12, C-13, C-14 and C-18. These correlations allowed us to place the C-26 methyl group at C-13 and C-27 methyl a group at C-14 respectively. The methyl protons at H-28 showed correlations with C-16, C-17, C-18 and C-22. These correlations allowed us to place C-28 methyl group at C-17. Finally, C-29 showed correlations with H-19, H-21 and H-30; H-30 showed correlations with H-19, H- 21 and H-29. C-20 showed correlations with H-19, H-21, H-29 and H-30; C-21 showed correlations with H- 22,H-29 and H-30 all these correlations supported the attachment of both C-29 and C-30 methyl groups to the same carbon at C-20 and the compound is an oleane type triterpenoid. The β orientation of the hydroxyl group at C-3 was confirmed by comparing the proton coupling between H-3 and H-4 and H-2 confirms the stereochemistry. The 13 C-NMR spectrum of 2 indicated the presence of 30 carbon resonances. The DEPT spectrum indicated the presence of six quaternary carbons, five CH carbons, eleven CH2 carbons and eight CH3 carbons; one of the carbons resonated at δ72.8 and indicative of presence of hydroxyl group.These data of compound 2 had good agreements with the reported data of 3β–friedelinol[18]. 3.1.3 Biological evaluation Pentacyclic triterpenoids (PTs) have been reported as naturally occurring active constituents having regulatory action on glucose metabolism by many biochemical mechanisms including inhibition of α- glucosidase and α-amylase. α-glucosidase is membrane bound enzyme integrated in the epithelium of the small intestine and is implicated for the last step of carbohydrate hydrolysis to create absorbable monosaccharide. Starch is the main dietary carbohydrate cause of glucose and diabetic complications depends upon the rate and extent of digestion of glucose from starch by the pancreatic α-amylase and intestinal α-glucosidase [19]. Disaccharides and oligosaccharides are broken down by the pancreatic α-amylase, followed by intestinal α-glucosidase enzyme which catabolizes disaccharides into glucose [20].The isolated terpenoidscompounds were evaluated for the inhibitory activity against α-glucosidase and α-amylase protein. Table 2 shows the inhibitory activity of terpenoidsagainst α-glucosidase and α-amylase in the percentage inhibition at 10-50µM concentration. Compound 2 (3β–friedelinol) inhibited the enzymes α-glucosidase and α- amylase with 15.57 and 36.35 percentage respectively and compound 1 (fredelin) inhibited the enzymes α- glucosidase and α-amylase with 10.4 and 23.04 percentage respectively. These terpenoids have shown better antidiabetic activity than marketed α-glucosidase inhibitor acarbose, which showed the inhibition of 9.46 percentages against both α-glucosidase and α-amylase. These results were in agreement to the in vitro α- glucosidase inhibition of terpenoids as per reported literature [10,21]. Antioxidants have been known to exert the protective effects against the oxidative damage and are associated with reduced risk of chronic diseases such as inflammation, diabetes, carcinogenesis and atherosclerosis [21,22,23]. Isolated PTs when investigated for their efficacy in scavenging free radicals through DPPH inhibition assay; Compound 2 (3β–friedelinol) was found to have better antioxidant activity, followed by compound 1 (fredeline) (Table 3). 3.1.4 Molecular Modeling In this research work, we also sought to give in silico evidence for the binding interaction of isolated compounds 1 (Fredeline) and 2 (3β–friedelinol) against α-glucosidase and α-amylase enzymes. The interactions were scored based on non-bonded interactions such as lipophilic pair term, hydrophobic enclosure reward,
  • 5. Isolation, in vitro antidiabetic, antioxidant activity and molecular docking studies of pentacyclic… DOI: 10.9790/3008-1062148154 www.iosrjournals.org 152 | Page hydrogen bonding, and electrostatic rewards (Table 4 and Table 5). As expected, compounds 1 and 2 were shown to have efficient antagonism of α-glucosidase and α-amylase enzymes. Acarbose is a tetrasaccharide possessing multiple free hydroxyl groups capable of forming numerous polar interactions with the active site of protein.While, compounds 1 (fredeline) and 2 (3β–friedelinol) being non polar in nature, exhibited enhanced hydrophobic interactions to inhibit α-glucosidase and α-amylase enzymes (Fig 1 and Fig 2). With α-glucosidase enzyme (Pdbcode: 3W37), compounds 1 (fredeline) and 2 (3β–friedelinol) possess hydrophobic interactions with Phe476, Trp432, Ile 358, Trp329, Phe601, Ala602 aminoacids. The hydrophobic interaction in acarbose were comparitively less (Phe 476, Trp432, Met470, Phe601) than polar hydrogen bonding interactions(His626, Asp357(2), Asp568, Arg552, Asp232). With α-amylase protein (Pdb code: 2QV4), the polar interaction are due to the hydroxyl group of 3β–friedelinol with Hie305 amino acid. The non-polar interactions of compounds 1 and 2 were found to be with Glu60, Trp58, Leu165, Val107, Ala106 aminoacids. Acarbose also possess similar interactions with additional hydrogen bonding with various amino acids (Asp300, Arg195, Glu233, Thr163, and Gln63). IV. Conclusion In summary, we report the isolation and characterization of Friedelin and 3β–Friedelinol pentacyclic terpenoids from the stem bark of Syzygium alternifolium. To our knowledge nobody has reported the identification of these two PTs from Syzygium alternifolium. The isolated compounds 1 (fredeline) and 2 (3β– friedelinol) PTs showed a showed significant inhibition of α-glucosidase, α-amylase and DPPH indicating the reduction in high blood glucose level as well as the reduction in the formation of ROS (Reactive Oxygen Species). Inhibition of these enzymes by compounds 1 (fredeline) and 2 (3β–friedelinol) was further supported by the molecular docking analysis. Therefore, these properties would be of therapeutic benefit in insulin resistant states. Future research will be carried out in this direction. Acknowledgements: We are grateful to the management of GokarajuRangaraju College of Pharmacy, Hyderabad and LailaNutraceuticals, Vijayawada for their support and cooperation. References [1]. Bazzano, L. Serdula, M.L.S. Prevention of type 2 diabetes by diet and lifestyle modification. J. Am. Coll. Nutr. 24, 2005, 310-319. [2]. Ahmed, D. Sharma, M. Mukerjee, A. Ramteke, P.W. Kumar, V. Improved glycemic control, pancreas protective and hepatoprotective effect by traditional poly-herbal formulation “QursTabasheer” in streptozotocin induced diabetic rats. BMC Complementary and Alternative Medicine, 2013, doi: 10.1186/1472-6882-13-10. [3]. Kumar, V. Ahmed, D. Gupta, P.S. Anwar, F. Mujeeb, M. Anti-diabetic, anti-oxidant and antihyperlipidemic activities of Melastomamalabathricum Linn. leaves in streptozotocin induced diabetic rats. BMC Complementary and Alternative Medicine 13,2013, 222. [4]. Ferrannini, E. Gastaldelli, A. Miyazaki, Y. Matsuda, M. Mari, A. DeFronzo, R. Beta-cell function in subjects spanning the range from normal glucose tolerance to overt diabetes: a new analysis. The Journal of Clinical Endocrinology and Metabolism, 90,2005, 493-500. [5]. Whiting, D.R. Guariguata, L. Weil, C. Shaw, J. IDF diabetes atlas: Global estimates of the prevalence of diabetes for 2011 and 2030. Diabetes Research and Clinical Practice, 94,2011, 311-321. [6]. Anjana, R.M. Pradeepa, R. Deepa, M. Datta, M. Sudha, V. Unnikrishnan, R. Prevalence of diabetes and prediabetes (impaired fasting glucose and/or impaired glucose tolerance) in urban and rural India: Phase I results of the Indian Council of Medical Research-India diabetes (ICMR-INDIAB) study. Diabetologia, 54,2011, 3022-3027. [7]. Hung, H.Y. Qian, K. Morris-Natschke, S.L. Hsu, C.S. Lee, K.H.Recent discovery of plant-derived anti-diabetic natural products.Natural Products Reports, 29, 2012, 580-606. [8]. Ahmed, D. Kumar, V. Sharma, M. Verma, A. Target guided isolation, in-vitro antidiabetic, antioxidant activity and molecular docking studies of some flavonoids from AlbizzialebbeckBenth. Bark. BMC Complementary and Alternative Medicine, 14,2014, 155. 9. Bailey, C. Day, C. Metformin: Its botanical background. Practical Diabetes International, 21,2004, 115-117. [9]. Ardiles, A.E. Rodriguez, A.G. Nunez, M.J. Perestelo, N.R. Studies of naturally occurring friedelanetriterpenoids as insulin sensitizers in the treatment of type 2 diabetes mellitus. Phytochemistry, 84,2012, 116-124. [10]. Ramesh, B. K. Abdul Nabi, S. Swapna, S. Apparao, C.Cinnamic acid as one of the antidiabetic active principle(s) from the seeds of Syzygium alternifolium. Food and Chemical and Toxicology, 50,2012, 1425-1431. [11]. Feng, J. Yang, X.W. Wang, R.F. Bio-assay guided isolation and identification of α-glucosidase inhibitors from the leaves of Aquilariasinensis. Phytochemistry,72,2011, 242-247. [12]. Wan, L.S. Chen, C.P. Xiao, Z.Q. Wang, Y.L. Min, Q.X. Yue, Y. Chen, J. In vitro and in vivo anti-diabetic activity of Swertiakouitchensis extract. Journal of Ethnopharmacology, 147, 2013, 622-630. [13]. Zhang, Y. Li, Q. Xing, H. Xuefeng, L. Zhao, L. Kankan, Q. Bi, K. Evaluation of antioxidant activity of ten compounds in different tea samples by means of an on-line HPLC–DPPH assay. Food Research International, 53,2013, 847-856. [14]. Raghavendra, N.M. Jyothsna, A. VenkateswaraRao, A. Subrahmanyam, C.V.S. Synthesis, pharmacological evaluation and docking studies of N-(benzo[d]thiazol-2-yl)-2-(piperazin-1- yl)acetamide analogs as COX-2 inhibitors. Bioorganic and Medicinal Chemistry Letters, 22,2012, 820-823. [15]. Raghavendra, N.M. Ramakrishna, K.Sirisha, V.Divya, P.VenkateswaraRao,A. Computer aided discovery of potential anti- inflammatory (s)-naproxen analogs as COX-2 inhibitors. Medicinal Chemistry,9, 2013, 553-559.
  • 6. Isolation, in vitro antidiabetic, antioxidant activity and molecular docking studies of pentacyclic… DOI: 10.9790/3008-1062148154 www.iosrjournals.org 153 | Page [16]. Hishashi, M. Toshiyuki, M. Yashiro, K.Johji, Y. Masayuki, Y. Antidiabetic principles of natural medicines. IV. Aldose reductase and α-glucosidase inhibitors from the roots of Salaciaoblonga Wall (Celastraceae): Structure of a new friedelane-type triterpene, kotalagenin 16- acetate. Chemical and Pharmaceutical Bulletin, 47,1999, 1725-1729. [17]. David Dako, C. Hanh Trinh, V.D. Wansi, J.D.New friedelane triterpenoids with antimicrobial activity from the stems of Drypetespaxii.Chemical and Pharmaceutical Bulletin, 57, 2009, 1119-1122. [18]. Dhital, S. Lin, AH-M. Hamaker, B.R. Gidley, M.J. Muniandy, A. Mammalian mucosal α-glucosidases coordinate with α-amylase in the initial starch hydrolysis stage to have a role in starch digestion beyond glucogenesis. PLoS One, 8, 2013, e62546. [19]. Kwon, Y.I. Apostolidis, E. Kim, Y.C. Shetty, K. Health benefits of traditional corn, beans and pumpkin: In vitro studies for hyperglycemia and hypertension management. Journal of Medicinal Food, 10,2007, 266-275. [20]. Tadera, K. Minami, Y. Takamatsu, K. Matsuoka, T. Inhibition of α-glucosidase and α-amylase by flavanoids. Journal of Nutritional Science and Vitaminology, 52,2006, 149-153. [21]. Liu, R.H. Adom, K.K. Antioxidant activity of grains. Journal of Agricultural and Food Chemistry, 50,2002, 6182-6187. [22]. Maria, S.M.R. Fabiano, A.N.F. Ricardo, E.A. Debrito, E.S. Free radical scavenging behavior of some north-east Brazilian fruits in a DPPH system. Food Chemistry, 114,2009, 693-695. [23]. Anderson, R.A. Evans, M.L. Ellis, G.R. Graham, J. Morris, K. Jackson, S.K. Lewis, M.J. Rees, A. Frenneaux, M.P.The relationship between post-prandial lipaemia, endothelial function and oxidative stress in healthy individuals and patients with type 2 diabetes.Atherosclerosis, 154, 2001, 475-483. Table 1: Structural details of isolated compounds from the bark of Syzygiumalternifolium Compound code Chemical structure IUPAC name Common names Compound 1 2 3 4 5 10 1 6 7 8 9 14 13 12 11 15 16 17 18 22 21 20 19 O 23 24 25 27 28 26 30 29 (4R,4aS,6bR,8aR,12bS,14aS)- octadecahydro- 4,4a,6b,8a,11,11,12b,14a- octamethylpicen-3(4H,6bH,14bH)- one Friedelin Compound 2 2 3 4 5 10 1 6 7 8 9 14 13 12 11 15 16 17 18 22 21 20 19 HO 23 24 25 27 28 26 30 29 (3S,4R,4aS,6bR,8aR,12bS,14aS)- docosahydro- 4,4a,6b,8a,11,11,12b,14a- octamethylpicen-3-ol 3β–Friedelinol Table 2: In vitro enzyme inhibition activity against α- glucosidase and α-amylase α-Glucosidase α-Amylase Compound IC50 value ± SEM IC50 value ± SEM Compound 1 10.40 ± 0.12** 23.04 ± 0.34* Compound 2 15.57 ± 0.23** 36.35 ± 0.21** Acarbose 09.46 ± 0.02 09.46 ± 0.02 SEM = Standard Error Mean; **p < 0.001, ***p < 0.0001 Table 3: Inhibition of radical scavenging activity by DPPH assay Samples IC50 value ± SEM Compound 1 20.5*** ± 0.39 Compound 2 30.5*** ± 0.22 Ascorbic acid 17.3 ± 1.10 SEM = Standard Error Mean; ***p < 0.0001 Table 4: Scoring Data (kcal/mol) of Glide-XP Docking of Schrodinger (Maestro 9.1) against α-amylase enzyme(PDB Code: 2QV4) Ligand Docking Scorea Lipophilic EvdWb H-Bondc Electrod RotPenale Acarbose -12.37 -3.87 -4.80 -1.95 0.10 Compound 2 -6.05 -3.27 -0.68 -0.13 0.00 Compound 1 -5.43 -2.93 0.00 -0.01 0.00 a An estimate of ligand- α-amylase binding energy; b Lipophilic van der Walls interactions of ligand- α-amylase complex; c Hydrogen-bonding term; d Electrostatic interactions of ligand- α-amylase complex; e Rotational penalty causing the decreased ligand-protein interactions.
  • 7. Isolation, in vitro antidiabetic, antioxidant activity and molecular docking studies of pentacyclic… DOI: 10.9790/3008-1062148154 www.iosrjournals.org 154 | Page Table 5: Scoring Data (kcal/mol) of Glide-XP Docking of Schrodinger (Maestro 9.1) against α- glucosidase enzyme (PDB Code: 3W37) Ligand Docking Scorea Lipophilic EvdWb H-Bondc Electrod RotPenale Acarbose -10.5 -1.94 -4.87 -2.00 0.00 Compound 1 -3.28 -1.17 0.00 -0.12 0.00 Compound 2 -3.57 -1.36 -0.70 -0.11 0.00 a An estimate of ligand- α-amylase binding energy; b Lipophilic van der Walls interactions of ligand- α-amylase complex; c Hydrogen-bonding term; d Electrostatic interactions of ligand- α-amylase complex; e Rotational penalty causing the decreased ligand-protein interactions. Fig. 1 Superimposition of compound 1, compound 2 and acarbose (line forms) in the binding site of α- amylase (PDB Code: 2QV4). Fig. 2 Overlay view of compounds 1, 2 and acarbose (stick forms) in α-glucosidase binding pocket surrounded by non-polar amino acids (space fill form) (PDB Code: 3W37).