International Conference on Natural Products 2015

Full paper

Synthesis and Evaluation of Benzimidazole Analogues Bearing Thiourea

Derivatives as α-Glucosidase Inhibitor

Nik Khairunissa’ Nik Abdullah Zawawiab, Muhammad Tahaab, Norizan Ahmata,b*, Nor Hadiani Ismaila,b, Noraishah Abdullaha
a
Faculty of Applied Science, UiTM Shah Alam, 40450 Shah Alam, Selangor D.E. Malaysia
b
Atta-ur-Rahman Institute for Natural Product Discovery, Universiti Teknologi MARA (UiTM), Puncak Alam Campus, 42300 Bandar Puncak Alam, Selangor
D. E. Malaysia
*Corresponding author: noriz118@salam.uitm.edu.my

Article history Abstract

Received : Benzimidazole and thiourea derivatives are of interest because of their widespread biological activities. A
Received in revised form : variety of synthesized benzimidazole and thiourea derivatives have been experimentally shown to possess
pharmacological activities including antimicrobial, antidiabetic, antitumor, antiinflammatory,
Accepted : antioxidative and others. Synthesis of target compounds begans with the synthesis of sodium metasulfite
adduct according to literature protocol. The resulting sulfite adduct was refluxed with 4,5-dimethyl-O-
Graphical abstract phenylenediamine in DMF for 6 hour to give the arylester substituted benzimidazole. The benzohydrazide
of benzimidazole was formed by refluxing arylester of benzimidazole with methanolic hydrazine hydrate.
The synthesis of the thiourea derivatives can be easily done with good yield by condensation of
benzimidazole benzoyl hydrazide with various phenyl isothiocyanate derivatives. Thus, we wish to
present on the synthesis and evaluation of some benzimidazole benzoyl hydrazide bearing thiourea
derivatives as α-glucosidase inhibitor.

Keywords: synthesis, benzimidazole, thiourea, α-glucosidase inhibitor

Abstrak

Derivatif benzimidazol dan thiourea menarik tumpuan kerana aktiviti biologinya yang meluas. Pelbagai
derivatif benzimidazol dan thiourea telah disintesis dan diuji secara eksperimental menunjukkan aktiviti
farmakologi

Kata kunci: sintesis, benzimidazol, thiourea

1.0 INTRODUCTION
Diabetes mellitus is a chronic metabolic disorder, characterized
by persistent hyperglycaemia resulting from defects in insulin
secretion, insulin action or both [1]. Type 2 diabetes mellitus
(T2DM), also called non-insulin-dependent diabetes mellitus,
is a common chronic disease that is characterized by
progressive β-cell dysfunction on a background of peripheral
insulin resistance and relative insulin deficiency [2]. Although
T2DM is prevalent mainly in adults, it currently occurs more
often in children and teenagers because of an increase in
obesity in children and adolescents [3]. Therefore, T2DM has
become an increasingly important public health issue
throughout the world [4].
α-Glucosidase inhibitors plays a key role in intestinal
carbohydrate digestion which are shown to control postprandial
hyperglycemia. Leading inhibitors such as acarbose aand
miglitol, are often reported to cause diarrhea and other intestinal
disturbances, with corresponding bloating, flatulence, cramping
and abdominal pain [5]. Hence, alternative α-glucosidase
inhibitors have been proposed as a potential therapeutic target
for drug discovery in the treatment of type 2 diabetes.
Benzimidazole nucleus is an important
pharmacophore with unique chemical and biological properties
[6-8]. Benzimidazoles have been found to possess anti-
inflammatory, antidiabetic, antispasmodic, antihistaminic,
analgesic, antimicrobial, antiproliferative, antitumor, anti-HIV-
RT, antiulcer, anticancer, anti-tubercular, and cycloxygenase
inhibitor activities [9-15]. Meanwhile, urea and thiourea
derivatives also exhibited many promising biological activities,
such as antimicrobial [16], antioxidant [17], antiviral [18],
antitumor [19], anti-inflammatory [20], antimalarial [21] and
antidiabetic [22] activities. Hence, the synergy effect from both
pharmacophore were hoping to enhance on the α-glucosidase
inhibitory activity.
2.0 EXPERIMENTAL
2.1 General
Melting point was taken on Buchi M-560 melting point
instrument and was uncorrected. IR spectra were recorded on a
Spectrum One FT-IR spectrometer (Perkin Elmer), using KBr
discs and values were signified in cm-1. The 1H NMR and 13C
NMR spectra were measured on Bruker 500 Ultrashield Plus
NMR (500 MHz) in DMSO-d6 as solvent, using
tetramethylsilane (TMS) as an internal standard, and chemical
shifts are expressed as ppm. ESI MS were determined on
Agilent 6330 Ion Trap using positive/negative mode at Faculty
of Pharmacy, UiTM Puncak Alam, Malaysia
2.2 Synthesis of methyl 4-(5,6-dimethyl-1H-
benzo[d]imidazol-2-yl)benzoate (a)
Sodium metasulfite adduct was synthesized according to
literature protocol [23]. Equimolar solution of sulphite adduct
and 4,5-dimethyl-O-phenylenediamine in DMF was refluxed for
6 hours. After the completion of reaction, it was poured into ice
water and the solid product (a) was filtered, dried and
crystallized.
Yield 96%. m.p. 208.9 °C; IR(KBr) (νmax, cm-1): 2954 (NH),
1720 (C=O), 1273, 1107 (C-O). 1H NMR (500 MHz, DMSO) δ:
2.33 (s, 6H, CH3-5,6-benzimidazole), 3.88 (s, 3H, OCH3-
benzoate), 7.39 (s, 2H, H-4,7-benzimidazole), 8.09 (d, J = 8.4
Hz, 2H, Ar), 8.27 (d, J = 8.4 Hz, 2H, Ar). 13C NMR (125 MHz,
DMSO) δ: 20.4 (CH3), 52.7 (OCH3), 126.8 (CH), 130.2 (CH),
130.4 (C), 131.7 (C), 135.0 (C), 149.5 (C), 166.3 (C). ESI MS
(m/z): 280.1 (M+).
2.3 Synthesis of 4-(5,6-dimethyl-1H-benzo[d]imidazol-2-
yl)benzohydrazide (b)
A solution of compound (a) (40 mmol) in methanol (50 ml) was
refluxed for 12 hours in the presence of hydrazine hydrate
(95%) mixture (10 ml). The reaction mixture was evaporated
and the residue was washed with water, filtered, dried, and
crystallized.
Yield 93%. m.p. 316.5 °C; IR(KBr) (νmax, cm-1): 3020 (NH),
1625 (C=N), 1560 (C-N), 1350 (N-N=C). 1H NMR (500 MHz,
DMSO) δ: 2.33 (s, 6H, CH3-5,6-benzimidazole), 4.56 (s, 2H,
NH2), 7.32 (s, 1H, H-7-benzimidazole), 7.46 (s, 1H, H-4-
benzimidazole), 7.98 (d, J = 7.5 Hz, 2H, Ar), 8.21 (d, J = 7.5
Hz, 2H, Ar), 9.88 (s, 1H, NH-hydrazide), 12.74 (s, 1H, NH-
benzimidazole). 13C NMR (125 MHz, DMSO) δ: 20.5 (CH 3),
111.9 (CH), 119.5 (CH), 126.5 (CH), 128.0 (CH), 130.7 (C),
132.1 (C), 133.2 (C), 134.1 (C), 134.2 (C), 143.0 (C), 150.0 (C),
165.8 (C). ESI MS (m/z): 280.1 (M+).
2.4 General procedure for the synthesis of benzimidazole
bearing thiourea derivatives (1-18)
Equimolar mixture of 4-(5,6-dimethyl-1H-benzo[d]imidazol-2-
yl) benzohydrazide (0.280 g, 0.01 mol) selected phenyl
isothiocanate (0.01 mol), in DMF (25 mL) was stirred overnight
at room temperature. The reaction mixture was filtered, dried
and crystallized.
N-(2-bromophenyl)-2-(4-(5,6-dimethyl-1H-benzo[d]imidazol-2-
yl)benzoyl)hydrazinecarbothioamide (1)
Yield 93%. m.p. 261.3°C; IR(KBr) (νmax, cm-1): 3265, 1676,
1529, 1396,1250, 1075, 1060, 852. 1H NMR (500 MHz, DMSO)
δ 10.69 (s, 1H), 9.85 (s, 1H), 9.64 (s, 1H), 8.26 (d, J = 8.3 Hz,
2H), 8.12 (d, J = 7.7 Hz, 2H), 7.66 (d, J = 7.9 Hz, 1H), 7.51 (s,
1H), 7.41 (d, J = 7.9 Hz, 3H), 7.21 (t, J = 7.7 Hz, 1H), 2.34 (s,
6H). 13C NMR (126 MHz, DMSO) δ 149.9, 133.9, 133.5, 132.9,
129.0, 128.1, 126.4, 20.5. EI-MS: 494.1 (M+1).
N-(2-chlorophenyl)-2-(4-(5,6-dimethyl-1H-benzo[d]imidazol-2-
yl)benzoyl)hydrazinecarbothioamide (2)
Yield 89%. m.p. 261.0°C; IR(KBr) (νmax, cm-1): 3269, 1675,
1616, 1458, 1250, 1128, 1059, 852. 1H NMR (500 MHz,
DMSO) δ 10.70 (s, 2H), 9.87 (s, 2H), 9.67 (s, 2H), 8.26 (d, J =
8.3 Hz, 4H), 8.12 (d, J = 7.9 Hz, 4H), 7.50 (d, J = 7.8 Hz, 3H),
7.45 – 7.33 (m, 6H), 7.28 (t, J = 7.3 Hz, 2H), 2.34 (s, 12H). 13C
NMR (126 MHz, DMSO) δ 149.9, 133.9, 133.4, 129.8, 129.0,
127.5, 126.3, 20.5. EI-MS: 450.1 (M+1).
2-(4-(5,6-dimethyl-1H-benzo[d]imidazol-2-yl)benzoyl)-N-(2-
fluorophenyl)hydrazinecarbothioamide (3)
Yield 91%. m.p. 196.6°C; IR(KBr) (νmax, cm-1): 3199, 1615,
1
1517, 1237, 854, 750. H NMR (500 MHz, DMSO) δ 10.70
(s, 1H), 9.89 (s, 1H), 9.65 (s, 1H), 8.26 (d, J = 8.3 Hz, 2H), 8.12
(d, J = 8.0 Hz, 2H), 7.40 (s, 2H), 7.32 – 7.17 (m, 3H), 2.34 (s,
6H). 13C NMR (126 MHz, DMSO) δ 149.8, 133.8, 133.5, 131.6,
128.9, 126.4, 124.4, 116.2, 20.5. EI-MS: 434.2 (M+1).
2-(4-(5,6-dimethyl-1H-benzo[d]imidazol-2-yl)benzoyl)-N-(2-
methoxyphenyl)hydrazinecarbothioamide (4)
Yield 93%. m.p. 267.4°C; IR(KBr) (νmax, cm-1): 3252, 1672,
1615, 1523, 1356, 1311, 1238, 1116, 1031, 852, 746. 1H NMR
(500 MHz, DMSO) δ 10.70 (s, 1H), 10.04 – 9.58 (m, 1H), 9.27

(s, 1H), 8.27 (d, J = 8.3 Hz, 2H), 8.09 (d, J = 8.2 Hz, 2H), 8.06 –
7.88 (m, 1H), 7.40 (s, 2H), 7.22 – 7.12 (m, 1H), 7.06 (d, J = 8.1
Hz, 1H), 6.95 (t, J = 7.7 Hz, 1H), 3.79 (s, 3H), 2.35 (s, 6H). 13C
NMR (126 MHz, DMSO) δ 149.8, 134.0, 133.3, 128.8, 128.4,
126.5, 120.3, 112.1, 56.3, 20.5. EI-MS: 446.2 (M+1).
N-(3,4-dichlorophenyl)-2-(4-(5,6-dimethyl-1H-
benzo[d]imidazol-2-yl)benzoyl)hydrazinecarbothioamide (5)
Yield 90%. m.p. 220.2°C; IR(KBr) (νmax, cm-1): 3277, 1644,
1609, 1596, 1459, 1264, 1132, . 1H NMR (500 MHz, DMSO) δ
10.67 (s, 1H), 10.09 – 9.88 (m, 2H), 8.27 (d, J = 8.4 Hz, 2H),
8.27 (d, J = 8.4 Hz, 2H), 8.11 (d, J = 8.2 Hz, 2H), 7.88 (s, 1H),
7.63 – 7.54 (m, 2H), 7.40 (s, 2H), 2.35 (s, 6H). 13C NMR (126
MHz, DMSO) δ 149.8, 140.0, 133.9, 133.3, 130.3, 128.9, 126.4,
20.4. EI-MS: 484.1 (M+1).
N-(3-bromophenyl)-2-(4-(5,6-dimethyl-1H-benzo[d]imidazol-2-
yl)benzoyl)hydrazinecarbothioamide (6)
Yield 92%. m.p. 212.1°C; IR(KBr) (νmax, cm-1): 3289, 1686,
1667, 1615, 1578, 1509, 1347, 1292, 1253, 1185, 1134. 1H
NMR (500 MHz, DMSO) δ 10.66 (s, 1H), 9.91 (s, 2H), 8.27 (d,
J = 8.3 Hz, 2H), 8.12 (d, J = 8.1 Hz, 2H), 7.79 (s, 1H), 7.56 (d,
J = 7.7 Hz, 1H), 7.40 (s, 2H), 7.33 (dt, J = 15.9, 7.9 Hz, 2H),
2.35 (s, 6H). 13C NMR (126 MHz, DMSO) δ 149.8, 141.5,
133.9, 133.4, 131.6, 130.3, 129.0, 126.4, 20.5. EI-MS: 494.1
(M+1).
2-(4-(5,6-dimethyl-1H-benzo[d]imidazol-2-yl)benzoyl)-N-(3-
fluorophenyl)hydrazinecarbothioamide (7)
Yield 90%. m.p. 208.6°C; IR(KBr) (νmax, cm-1): 3263, 1667,
1614, 1530, 1488, 1449, 1357, 1265, 1226, 1146. 1H NMR (500
MHz, DMSO) δ 10.66 (s, 1H), 9.92 (s, 2H), 8.28 (d, J = 8.2 Hz,
2H), 8.12 (d, J = 8.1 Hz, 2H), 7.54 (s, 1H), 7.44 – 7.33 (m, 4H),
6.99 (t, J = 7.5 Hz, 1H), 2.35 (s, 6H). 13C NMR (126 MHz,
DMSO) δ 149.8, 141.6, 141.5, 133.9, 133.4, 131.6, 128.9,
126.4, 20.4. EI-MS: 434.1 (M+1).
2-(4-(5,6-dimethyl-1H-benzo[d]imidazol-2-yl)benzoyl)-N-(3-
methoxyphenyl)hydrazinecarbothioamide (8)
Yield 94%. m.p. 218.8°C; IR(KBr) (νmax, cm-1): 3271, 1682,
1602, 1542, 1494, 1356, 1311, 1248, 1040. 1H NMR (500 MHz,
DMSO) δ 10.61 (s, 1H), 9.81 (s, 1H), 9.73 (s, 1H), 8.26 (d, J =
8.3 Hz, 2H), 8.11 (d, J = 8.2 Hz, 2H), 7.40 (s, 2H), 7.24 (dd, J =
18.1, 10.0 Hz, 2H), 7.08 (d, J = 7.9 Hz, 1H), 6.75 (dd, J = 8.2,
1.9 Hz, 1H), 3.76 (s, 3H), 2.35 (s, 6H). 13C NMR (126 MHz,
DMSO) δ 149.8, 140.9, 133.8, 133.5, 128.9, 126.4, 55.6, 20.4.
EI-MS: 446.2 (M+1).
2-(4-(5,6-dimethyl-1H-benzo[d]imidazol-2-yl)benzoyl)-N-(3-
nitrophenyl)hydrazinecarbothioamide (9)
Yield 93%. m.p. 214.5°C; IR(KBr) (νmax, cm-1): 3259, 1671,
1615, 1529, 1345, 1278, 1145, 1052. 1H NMR (500 MHz,
DMSO) δ 10.71 (s, 1H), 10.10 (dd, J = 26.2, 1.5 Hz, 2H), 8.50
(s, 1H), 8.27 (d, J = 8.3 Hz, 2H), 8.12 (d, J = 8.1 Hz, 2H), 8.08 –
7.93 (m, 2H), 7.63 (t, J = 8.2 Hz, 1H), 7.40 (d, J = 1.0 Hz, 2H),
2.35 (s, 6H). 13C NMR (126 MHz, DMSO) δ 149.8, 141.1,
139.4, 134.0, 133.3, 133.2, 129.0, 126.4, 20.5. EI-MS: 461.1
(M+1).
N-(4-bromophenyl)-2-(4-(5,6-dimethyl-1H-benzo[d]imidazol-2-
yl)benzoyl)hydrazinecarbothioamide (10)
Yield 86%. m.p. 227.9°C; IR(KBr) (νmax, cm-1): 3323, 1672,
1612, 1528, 1489, 1353, 1290, 1255, 1190, 1136, 1073, 1013.
1
H NMR (500 MHz, DMSO) δ 12.79 (s, 1H), 10.66 (s, 1H),
9.98 – 9.80 (m, 2H), 8.26 (d, J = 8.3 Hz, 2H), 8.11 (d, J = 8.1
Hz, 2H), 7.53 (d, J = 8.5 Hz, 2H), 7.48 (s, 2H), 7.40 (s, 2H),
2.34 (s, 6H).
13
C NMR (126 MHz, DMSO) δ 149.8, 139.2, 133.9, 133.4,
131.3, 129.0, 126.3, 20.5. EI-MS: 494.1 (M+1).
N-(4-chlorophenyl)-2-(4-(5,6-dimethyl-1H-benzo[d]imidazol-2
yl)benzoyl)hydrazinecarbothioamide (11)
Yield 88%. m.p. 213.2°C; IR(KBr) (νmax, cm-1): 3312, 1694,
1613, 1528, 1354, 1291, 1257, 1190, 1137, 1094, 1016. 1H
NMR (500 MHz, DMSO) δ 12.76 (s, 1H), 10.63 (s, 1H), 9.88(s,
1H), 9.82 (s, 1H), 8.26 (d, J = 8.4 Hz, 2H), 8.10 (d, J = 8.3 Hz,
2H), 7.53 (d, J = 5.7 Hz, 2H), 7.39 (d, J = 8.7 Hz, 4H), 2.35 (s,
6H). 13C NMR (126 MHz, DMSO) δ 149.8, 138.8, 133.9, 133.4,
128.9, 128.4, 126.3, 20.5. EI-MS: 450.2 (M+1).
2-(4-(5,6-dimethyl-1H-benzo[d]imidazol-2-yl)benzoyl)-N-(4-
fluorophenyl)hydrazinecarbothioamide (12)
Yield 93%. m.p. 203.1°C; IR(KBr) (νmax, cm-1): 3280, 1672,
1615, 1521, 1363, 1290, 1260, 1224, 1147, 1058. 1H NMR (500
MHz, DMSO) δ 10.62 (s, 1H), 9.83 (s, 1H), 9.75 (s, 1H), 8.26
(d, J = 8.4 Hz, 2H), 8.11 (d, J = 8.3 Hz, 2H), 7.47 (s, 2H), 7.40
(s, 2H), 7.17 (t, J = 8.8 Hz, 2H), 2.35 (s, 6H). 13C NMR (126
MHz, DMSO) δ 149.8, 136.1, 133.8, 133.5, 129.0, 126.3, 115.2,
115.0, 20.5. EI-MS: 434.2 (M+1).
2-(4-(5,6-dimethyl-1H-benzo[d]imidazol-2-yl)benzoyl)-N-(4-
methoxyphenyl)hydrazinecarbothioamide (13)
Yield 83%. m.p. 215.2°C; IR(KBr) (νmax, cm-1): 3256, 1668,
1614, 1509, 1292, 1251, 1043. 1H NMR (500 MHz, DMSO) δ
10.60 (s, 1H), 9.80 (s, 1H), 9.73 (s, 1H), 8.26 (d, J = 8.3 Hz,
2H), 8.11 (d, J = 8.3 Hz, 2H), 7.40 (s, 2H), 7.18 (dt, J = 62.4,
8.0 Hz, 3H), 6.75 (dd, J = 8.2, 2.0 Hz, 1H), 3.76 (s, 3H), 2.35 (s,
6H). 13C NMR (126 MHz, DMSO) δ 149.8, 140.9, 133.9, 133.5,
129.0, 126.3, 55.6, 20.5. EI-MS: 446.1 (M+1).
2-(4-(5,6-dimethyl-1H-benzo[d]imidazol-2-yl)benzoyl)-N-(4-
nitrophenyl)hydrazinecarbothioamide (14)
Yield 91%. m.p. 196.5°C; IR(KBr) (νmax, cm-1): 3167, 1592,
1508, 1465, 1323, 1248, 1176, 1112, 1001. 1H NMR (500 MHz,
DMSO) δ 10.75 (s, 1H), 10.17 (s, 1H), 8.27 (d, J = 9.2 Hz, 2H),
8.22 (d, J = 9.1 Hz, 2H), 8.11 (t, J = 8.6 Hz, 2H), 7.95 (d, J =
8.5 Hz, 2H), 7.41 (s, 2H), 2.34 (s, 6H). 13C NMR (126 MHz,
DMSO) δ 149.8, 133.9, 133.3, 131.6, 128.9, 126.5, 20.5. EI-
MS: 461.2 (M+1).
2-(4-(5,6-dimethyl-1H-benzo[d]imidazol-2-yl)benzoyl)-N-(4-
(trifluoromethyl)phenyl)hydrazinecarbothioamide (15)
Yield 90%. m.p. 220.1°C; IR(KBr) (νmax, cm-1): 3321, 1672,
1613, 1533, 1351, 1259, 1164, 1125, 1070, 1018. 1H NMR (500
MHz, DMSO) δ 10.67 (s, 1H), 9.97 (bs, 2H), 8.26 (d, J = 8.3
Hz, 2H), 8.11 (d, J = 8.1 Hz, 2H), 7.80 (s, 2H), 7.70 (d, J = 8.5
Hz, 2H), 7.40 (d, J = 1.2 Hz, 2H), 2.35 (s, 6H). 13C NMR (126
MHz, DMSO) δ 149.8, 143.6, 133.9, 133.4, 128.9, 126.4, 100.0,
20.5. EI-MS: 484.1 (M+1).
2-(4-(5,6-dimethyl-1H-benzo[d]imidazol-2-yl)benzoyl)-N-(m-
tolyl)hydrazinecarbothioamide (16)
Yield 86%. m.p. 209.7°C; IR(KBr) (νmax, cm-1): 3251, 1666,
1615, 1542, 1487, 1440, 1355, 1266, 1228, 1143, 1056. 1H
NMR (500 MHz, DMSO) δ 10.59 (s, 1H), 9.76 (s, 1H), 9.66 (s,
1H), 8.25 (d, J = 8.4 Hz, 2H), 8.10 (d, J = 8.4 Hz, 2H), 7.32 (dd,
J = 34.5, 26.8 Hz, 4H), 7.22 (t, J = 7.7 Hz, 1H), 6.99 (d, J = 7.5
Hz, 1H), 2.33 (s, 6H), 2.31 (s, 3H). 13C NMR (126 MHz,
DMSO) δ 149.8, 139.6, 133.8, 133.5, 128.9, 126.3, 21.4, 20.5.
EI-MS: 430.2 (M+1).
2-(4-(5,6-dimethyl-1H-benzo[d]imidazol-2-yl)benzoyl)-N-(o-
tolyl)hydrazinecarbothioamide (17)
Yield 86%. m.p. 225.7°C; IR(KBr) (νmax, cm-1): 3265, 1668,
1617, 1509, 1341, 1286, 1249, 1231, 1135, 1002. 1H NMR (500
MHz, DMSO) δ 12.75 (s, 1H), 10.61 (s, 1H), 9.62 (s, 1H), 9.57
(s, 1H), 8.24 (d, J = 8.5 Hz, 2H), 8.10 (d, J = 8.3 Hz, 2H),
7.45(s, 1H), 7.33 (s, 1H), 7.35 – 6.90 (m, 4H), 2.34 (s, 12H),
2.22 (s, H). 13C NMR (126 MHz, DMSO) δ 149.9, 133.8, 133.6,
130.5, 129.0, 127.1, 126.3, 20.5, 18.2. EI-MS: 430.2 (M+1).
2-(4-(5,6-dimethyl-1H-benzo[d]imidazol-2-yl)benzoyl)-N-(p-
tolyl)hydrazinecarbothioamide (18)
Yield 92%. m.p. 206.2°C; IR(KBr) (νmax, cm-1): 3318, 1672,
1614, 1530, 1357, 1291, 1256, 1135, 1017. 1H NMR (500 MHz,
DMSO) δ 10.58 (s, 1H), 9.76 (s, 1H), 9.63 (s, 1H), 8.25 (d, J =
8.3 Hz, 2H), 8.10 (d, J = 8.3 Hz, 2H), 7.40 (s, 2H), 7.34 (d, J =
7.2 Hz, 2H), 7.15 (d, J = 8.1 Hz, 2H), 2.35 (s, 6H), 2.30 (s, 3H).
13
C NMR (126 MHz, DMSO) δ 149.8, 137.2, 133.8, 133.5,
129.0, 126.3, 21.0, 20.5. EI-MS: 430.1 (M+1).
derivatives using Baker’s yeast α-glucosidase enzyme.
Compounds 1-18 exhibited a varying degree of α-glucosidase
inhibitory activity with IC50 values between 35.83±0.66 -
297.99±1.20 μM when compared with standard acarbose (IC50 =
774.5±1.94 μM). However, only compound 10 and 14 showed
significant inhibitory effects with IC50 = 50.57±0.81 and
35.83±0.66 μM, respectively than the rest of the series (Table
1). It was observed that α-glucosidase inhibitors mainly depend
upon three parameters: the functional groups on substituted
phenyl moiety, and the number and the position of the
functional groups on the phenyl ring. As shown in Table 1, both
compounds having electron withdrawing group (Br and NO 2) at
para-position.
NH2
O O Na2S2O5, MeOH O OH NH2
+
O H H2O 80%, rt O SO3Na

2.5 Baker’s Yeast α-glucosidase inhibition assay DMF, reflux

6 hours
The enzyme inhibition was evaluated according to the method
previously reported by Shibano et al. (1997) [24]. with minor N O Hydrazine N O
modification. Test sample (10 μl) was premixed with 95 μl of 50 N HN NH2 MeOH N O
mM phosphate buffer (pH 6.8), 25 μl of α-glucosidase solution H H
(a stock solution of 1 mg/ml in phosphate buffer, was diluted to RSCN,
0.0625 U/ml with the same buffer just before the assay) and pre- THF, rt
incubated at 37°C for 10 min. The reaction was initiated with 4 3
2' 3'
the addition 25 μl of 5 mM PNPG (dissolve 1.5 mg in 1 ml of N O

phosphate buffer) and absorbance at time 0 minutes was N HN NH

measured. The reaction mixture was then incubated at 37°C for 7 H 6' 5'
S
1
30 min and absorbance at time 30 minutes was measured. For (1-18) HN
R
negative control, the test samples were replaced with 10 μl of
Scheme 1 Synthesis of benzimidazole thiourea derivatives 1-18
DMSO and acarbose was used as positive control. The
enzymatic hydrolysis of the substrate was monitored based on 4.0 CONCLUSION
the amount of p-nitrophenol released in the reaction mixture by
observation at 405 nm using a microplate reader. All
experiments were carried out in triplicate and the results are Acknowledgement
expressed as the mean ± S.E.M of three determinations. The
percentage (%) inhibition of α-glucosidase inhibitory activity The authors would like to acknowledge the Ministry of Higher
was calculated using the equation: Education (MOHE) and Universiti Teknologi MARA for the
30 min
– A0 min)control – (A30 min– A0 min) exp financial support under RAGS grant 600-RMI/RAGS/5/3/
% Inhibition = (A × 100%
(2/2012).
(A30 min – A0 min)control
References
2.6 Statistical analysis
[1] Muhlisah, F., 2007. Tanaman Obat Keluarga TOGA. Penebar
Swadaya., 1, 46-47.
[2] Harmanto, N., 2003. Conquering Disease in Unison with Mahkota
3.0 RESULTS AND DISCUSSION Dewa, Ir. Harmantop (Ed.), pg 14, PT. Mahkota Dewa Indonesia,
North Jakarta.
[3] Madhuri, S. and Pandey, G., 2009. Some Anticancer Medicinal Plants
tabIdentification of potential α-glucosidase inhibitors were done of Foreign Origin. Current Science, 96 (6), 779- 783.
by in vitro screening of 18 benzimidazole bearing thiourea

Table 1 α-Glucosidase inhibitory activity of benzimidazole thiourea derivatives 1-18

No Substituent IC50±SEMa (μM) No Substituent IC50±SEMa (μM) No Substituent IC50±SEMa (μM)

7 13
1 2'' Br 129.45±0.46 246.83±98 305.25±1.19

3''
F 4''
OCH3
 8 14
2 2'' Cl 284.47±1.10 71.82±0.28 35.83±0.66

3''
OCH3 4''
NO2
9 15
3 2'' F 230.68±0.92 117.26±0.44 163.39±0.59

3''
NO2 4''

F F
F
10 16
4 2'' OCH3 345.65±1.35 50.57±0.81 218.84±0.88

3''

4''
Br
11 17
5 61.93±0.22 186.68±0.71 2'' *NA

3''
4'' Cl 4''
Cl Cl
12 18
6 177.99±0.63 276.82±0.64 297.99±1.20

3''
Br 4'' 4''
F
*NA= not active 19 Acarbose 774.5±1.94
a
SEM is the standard error of the mean, Acarbose is standard inhibitor for α-glucosidase.