Antiradical, antimicrobial and enzyme inhibition evaluation of sulfonamide derived esters; synthesis, X-Ray analysis and DFT studies

Accepted Manuscript Antiradical, Antimicrobial and Enzyme Inhibition Evaluation of Sulfonamide Derived Esters; Synthesis, X-Ray Analysis and DFT Studies

Muhammad Danish, Ayesha Bibi, Khola Gilani, Muhammad Asam Raza, Muhammad Ashfaq, Muhammad Nadeem Arshad, Abdullah Mohamed Asiri, Khurshid Ayub PII:

S0022-2860(18)30943-8

DOI:

10.1016/j.molstruc.2018.07.116

Reference:

MOLSTR 25521

To appear in:

Journal of Molecular Structure

Received Date:

01 June 2018

Accepted Date:

31 July 2018

Please cite this article as: Muhammad Danish, Ayesha Bibi, Khola Gilani, Muhammad Asam Raza, Muhammad Ashfaq, Muhammad Nadeem Arshad, Abdullah Mohamed Asiri, Khurshid Ayub, Antiradical, Antimicrobial and Enzyme Inhibition Evaluation of Sulfonamide Derived Esters; Synthesis, X-Ray Analysis and DFT Studies, Journal of Molecular Structure (2018), doi: 10.1016/j. molstruc.2018.07.116

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ACCEPTED MANUSCRIPT

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Antiradical, Antimicrobial and Enzyme Inhibition Evaluation of Sulfonamide Derived

2

Esters; Synthesis, X-Ray Analysis and DFT Studies

3

Muhammad Danisha*, Ayesha Bibia, Khola Gilania, Muhammad Asam Razaa, Muhammad

4

Ashfaqa, Muhammad Nadeem Arshadb, Abdullah Mohamed Asirib*, Khurshid Ayubc aDepartment

5 6

bChemistry

Department, Faculty of Science, King Abdulaziz University, Jeddah 21589, Saudi Arabia

7 8

of Chemistry, University of Gujrat, Gujrat 50700 Pakistan

cDepartment

of Chemistry, COMSATS Institute of Information Technology, Abbottabad, KPK,

9

Pakistan, 22060

10

Authors for correspondence:[email protected], [email protected]

11 12

Abstract

13

Two carboxylate esters (methyl: (I) and ethyl: (II)) of 4-{(4-methylphenylsulfonamido)-

14

methyl}cyclohexanecarboxylic acid (sulfonamide) were synthesized and characterized by FTIR

15

and X-ray crystallography. DFT studies were conducted in order to optimize the structures using

16

Gaussian software which confirmed the bond angels and bond lengths obtained from single

17

crystal analysis. Both Compounds (I and II) were evaluated for their biological studies viz;

18

antioxidant activity (DPPH ), enzyme inhibition activity (esterase and proteases), antibacterial

19

(Halomonas halophila, Halomonas salina, Shigella sonnei, Bacillus subtilis, Chromohalobacter

20

salexigens, Chromohalobacter israelensis, Staphylococcus aureus, Escherichia coli and

21

Klebsiella pneumoniae) and anti-fungal (Aspergillus niger and Alternaria alternata). Results

22

depicted that II is more active as compared to I in antioxidant and esterases while I is more

23

potent against protease while moderate results were shown by both.

24

Keywords: Antioxidant; DFT; Enzyme Inhibition; Ester;. Sulfonamide. .

25 26 27

1.

Introduction

28

Sulfonamides have been found to show broad pharmacological profile. They are stable in human

29

body and being used for the treatment of various diseases i.e. tumor, diabetes and other major 1|Page

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pathogens. Moreover, they are used in agriculture field as well as insecticides and herbicides.

31

They are less toxic as compared to other drugs and are scalable

32

that minor variation in the structure of sulfonamide gives vast range of applications both

33

qualitatively as well as quantitatively. Sulfonamides have been proved to be wonderful drugs as

34

they serve the humanity meritoriously for health e.g. as antitumor, antibacterial, antifungal and

35

inhibition against lipoxygenase enzyme [4-7]. As β3 adrenergic agonist, they serve for the

36

treatment of obesity and type 2 diabetes [8-10]. The biological potential of the sulfonamides

37

depends to which way, they attach to their respective receptor or enzyme. This aptitude of

38

binding depends upon proton-ligand complex of the sulfonamides [11]. After the discovery of

39

sulfanilamide, so many chemical alteration have been done and evaluated their therapeutic

40

results.

41

following different synthetic schemes. These syntheses have resulted in the finding of new

42

agents with changeable pharmacological properties [12]. Density functional theory (DFT) has

43

long been renowned as a best tool in the understanding of organic complex previous methods

44

used in the past. Detailed analyses on the performance of various DFT methods have been

45

carried out predominantly for the optimized geometry or structure [13]. Keeping in view such

46

therapeutic applications, new esters derived from sulfonamides were synthesized and

47

characterized. Evaluation of these synthesized compounds for their pharmacological profile

48

involving enzyme inhibition, bactericidal, fungicidal and anti-oxidant potential is also part of this

49

research task.

[1-3]. It has been reported

Up-to date greater than 20,000 sulfanilamide derivatives have been synthesized

50 51

2.

Experimental

52

Chemical for this research task were purchased from Alfa Aesar and Merck (UK). Solvents used;

53

Methanol, Ethanol and Dimethyl Sulfoxide of analytical grade were purchased from Merck

54

(UK). Perkin-Elmer System 100 FT-IR spectrophotometer was used for IR spectral data (4000-

55

400 cm-1) while X-ray diffraction analysis was done on Atlas diffractometer.

56

2.1

57

(I) and Ethyl-4-{(4-methylphenylsulfonamido)methyl} cyclohexanecarboxylate (II)

58

10 mL alcohol (methanol and ethanol) was taken in 100 mL round bottom flask and acidified

59

with 1 mL of conc. H2SO4, followed by the addition of 0.5 gram of 4-{(4-

60

methylphenylsulfonamido)methyl}cyclohexanecarboxylic acid. The mixture was refluxed for

Synthesis of Methyl-4-{(4-methylphenylsulfonamido)methyl} cyclohexanecarboxylate

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about 6 hours and stirred overnight at room temperature. After stirring it was concentrated at

62

room temperature via slow evaporation until crystalline product obtained. H3C O

CH3OH

S O

NH

H3C

OCH3

(I)

O S O

O

NH OH

H3C

O

O

C2H5OH

S O

NH OC 2 H5

(II)

63

O

64

2.2

Characterization of the Synthesized Esters

65

Both compounds were characterized on the basis of FTIR and single XRD analysis.

66

2.3

67

Quantum chemical calculations were performed with Gaussian 09 (Revision C.01) [14]. The

68

results are visualized with Gauss View 5.0. The geometries of the compounds are optimized

69

without any symmetry constraints using the hybrid functional B3LYP method 6-31G(d,p) basis

70

set [15]. B3LYP method consists of Becke’s three-parameter (B3) hybrid exchange functional in

71

conjunction with the correlation functional of Lee Yang and Parr (LYP) [16,17]. The basis set

72

chosen contains polarization functions on all atoms. The B3LYP method of DFT is quite reliable

73

for the prediction of geometric and electronic properties of neutral and charged species ranging

74

from simple molecular to polymer structures [18-26]. For optimization, the input geometries are

75

taken from the crystal structure (where available) in order to better match with the

76

experimentally obtained structures. Frequency calculations are also performed at the same level

77

in order to confirm these structures as true minima (absence of an imaginary frequency).

78

2.4

Biological Activities

79

2.4.1

Antimicrobial Assay

DFT studies

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Disc diffusion method was used to measure antimicrobial activity of synthesized compounds

81

against nine bacterial strains; Halomonas halophila, Halomonas salina, Shigella sonnei, Bacillus

82

subtilis, Chromohalobacter salexigens, Chromohalobacter israelensis, Staphylococcus aureus,

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Escherichia coli, Klebsiella pneumoniae and two fungal strains (Alternaria alternata and

84

Aspergillus niger) according to procedure of Shahid et al., (2009) [27]. Prepared the growth

85

medium, autoclaved at 121oC and transferred 30 mL of this solution to the petri dishes. After

86

solidification of medium, corresponding strain was used to seed medium. 20 µL (5 mg/mL)

87

sample solution was loaded on each disc. Kanamycin, ampicillin and streptomycin were used as

88

reference drugs for bacteria while fungivin was used as antifungal drug. The plates were

89

incubated at 37 oC (24 hours) and 25 oC (48 hours) for bacterial and fungal respectively.

90

2.4.2

91

The methodology of Shahwar et al., (2010) was used to measure the anti-oxidant activity by

92

using radical (2-2’-dienyl-1-picrylhydrazyl) [28]. DPPH solution was prepared in methanol with

93

concentration level of 0.0025g/mL. In first step, added 100µL of each of compound’s sample

94

solution into 2mL DPPH solution in test tube and allowed to stand in darkness for 30 minutes.

95

The absorbance of mixture was recorded at 517 nm keeping methanol was used as blank while

96

Gallic acid was used as standard. % age inhibition was calculated using formula given below.

97

% Inhibition 

98

2.4.3

99

Esterases inhibitory activity of synthesized compounds against acetyl cholineesterase (AChE)

100

and butyrylcholine esterase was determined by method of Ellman et al., (1961) along some

101

modifications [29]. 100 μL test compound (5 mg/mL) was mixed with 100 μL enzyme (AChE

102

and BChE) and incubate at 37 °C for 10 minutes. After incubation 0.5 mL buffer (50 mM), 50

103

μL DTNB followed by addition of 50 μL substrate acetylthiocholine iodide and

104

butyrylthiocholine iodide for AChE and BChE respectively. After 30 minutes of incubation at 37

105

°C, the absorbance was measured at 410 nm using UV/VIS spectrophotometer. All experiments

106

were carried out with their respective controls in triplicate. The %age inhibition was calculated

107 108

by the formula mentioned in antioxidant assay.

109

2.4.4

Antioxidant Assay

Absorbance (blank) - Absorbance (test)  100 Absorbance (blank)

Esterases Inhibitory Activity

α-Chymotrypsin Protease Assay 4|Page

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Protease inhibition study of compounds was done by using methodology of Raza, et al., (2013)

111

with minor modifications [30]. In first step, added 100 µL of test sample as well as enzyme,

112

shaked and waited for 10 minutes. 0.5 mL of tris-buffer was added in this solution mixture

113

followed by addition of substrate. Incubation of mixture was done at 37 oC for 30 minutes.

114

Added 0.5 mL distilled water in this solution and recorded the absorbance at 410 nm with UV-

115

VIS spectrophotometer. The %age inhibition was calculated by the formula mentioned in

116

antioxidant assay.

117

3.

118

The sulfonamide was reacted in acidic medium with methanol (I) and ethanol (II) on stirring

119

which resulted white precipitates. The products (I and II) were crystallized in ethanol and

120

characterized with FT-IR and single crystal XRD techniques. In infrared spectra of compounds

121

(I) and (II), the disappearance of broad peak around 3000 cm-1 indicated the deprotonation of

122

acid and formation of ester. Both esters showed the sharper peak in range of 3226 cm-1 (I) to

123

3298 cm-1 (II) indicating the presence of NH group [31]. CH of aliphatic group showed the peak

124

in range of 2922 cm-1 (I) to 2931cm-1 (II). SO2 group gave sharp peaks of the asymmetric and

125

symmetric stretching frequencies in range of 1314 - 1315 cm-1 and 1140 -1146 cm-1 respectively.

126

The stretching frequencies for carboxylate group (COO) were given in the range of 1718 cm-1 (I)

127

to 1720 cm-1 (II) for both compounds showing the confirmation of esters moiety in compounds

128

[32].

Result and Discussion

129

The information about the spatial arrangements of molecules (I and II) in the unit cell is

130

very important for their further physico-chemical properties. Samples of crystallized material

131

were screened out under microscope and glued over a glass fiber tip immersed in a wax on

132

copper rod with magnetic base. This holder was mounted on Agilent Super Nova (Dual source)

133

Agilent Technologies Diffractometer, equipped with graphite-monochromatic Cu/Mo Kα

134

radiation for data collection. The data collection was accomplished using CrysAlisPro software

135

at 296 K under the Mo Kα radiation [33]. The structure solution was performed using SHELXS–

136

97 and refined by full–matrix least–squares methods on F2 using SHELXL–97, in-built with

137

WinGX [34,35]. All non–hydrogen atoms were refined anisotropically by full–matrix least

138

squares methods [34]. Figures were drawn using PLATON and ORTEP-3 [36,37].

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All the hydrogen atoms attached to the aromatic carbon atoms were positioned geometrically and

140

treated as riding atoms with C–H = 0.93 Å and Uiso(H) = 1.2 Ueq(C) carbon atoms. The N-H =

141

0.85(1) Å, hydrogen atom was located with difference fourier map and refined using the DFIX

142

restraint with Uiso (H) = 1.2 Ueq(O). The methyl and methylene hydrogen atoms were also

143

positioned geometrically with C–H = 0.96 Å and Uiso(H) = 1.5 Ueq(C) for methyl group and C–

144

H = 0.97 Å and Uiso(H) = 1.2 Ueq(C) for methylene hydrogen atoms. The Van der Waals’

145

interactions among the molecules produce extra stability of crystal structures. The compound (I)

146

was crystallized with the emphasis to know internal geometry and Van der Waals’ interaction of

147

the molecules in their crystal structures. The study is in continuation of already reported crystal

148

structures of sulfonamides molecules by our group [38,39]. The presented molecule (Figure 1)

149

was crystallized with monoclinic crystal system and P2 1/n space group Table 1. Hydrogen

150

bonding were shown in Table 2 while selected bond lengths and bond angles are provided in

151

Table 3 and 4 respectively.

152

153 154

Figure 1: ORTEP diagram of molecule I where thermal ellipsoids were drawn at 50%

155

probability level

156 157

The cyclohexane ring adopted the chair shape conformation and the root mean square (r. m. s)

158

deviation for the fitted atoms of this ring is 0.2308 Å. The dihedral angles between the planes of

159

cyclohexane atoms and aromatic ring is 21.55 (1)º which is lesser than its parent molecule 4-{(4-

160

methylbenzenesulfonamido)methyl}cyclohexanecarboxylic acid.23 The puckering parameters for

161

the cyclohexane rings are QT = 0.565 (2) Å, θ = 177.8 (5)°, φ = 188.8 (6)° [40]. Methyl-ester

162

group is twisted with the dihedral angle of 53.20 (1) º with respect to cyclohexane ring. A very 6|Page

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beautiful network produced through the intermolecular hydrogen bonding which connects the

164

molecules to form the infinite chains along c-axis [0 0 1] as shown in Figures 2 & 3. Atoms N(1)

165

and C(12) in the molecule situated at (x, y, z) act as hydrogen bond donor via H(1N) and H12, to

166

atoms O(4) and O(1) respectively at (x, y, z-1). Both the interactions produced seventeen

167

membered ring motif R22 [41].

168 169 170 171

Figure 2: Unit Cell Diagram of I, showing intermolecular hydrogen bonding interactions

172

using dashed lines

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173 174

Figure 3: Formation of infinite one-dimensional chains along c-axes through hydrogen

175

bonding

176

The molecule II was crystalized in orthorhombic crystal system and unit cell parameters were a =

177

5.3382(3) Å, b = 9.5791(9) Å, c = 36.078(3) Å, V = 1844.8(3) Å3. The data was collected at T =

178

296.15 K with space group P212121 (no. 19) and Z = 4. The final wR2 was 0.1647 (all data)

179

and R1 was 0.0634 (I > 2\s(I)). In the crystal structure of compound II, the O-S-O angle around

180

the S atom is 120.6(2)º giving rise to formation of distorted tetrahedral geometry, where the

181

corner of tetrahedron are being occupied by the C1, N1, O1 and O2 atoms. The dihedral angle

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between the aromatic ring and plane produced through the fitted atoms of cyclohexane ring is

183

62.086 (2)º. The root mean square (r.m.s) deviation for the cyclohexane ring observed is 0.2318

184

Å and puckering parameters observed are QT = 0.568 (2) Å, θ = 176.99 (5)°, φ = 6.807 (2)°. The

185

ethyl group is disordered over two positions with the site occupancy of 0.73 and 0.27 for major

186

and minor components. Infinite chains observed, produced through the hydrogen bonding

187

interaction between the N-H and SO2 of sulfonamide group. Atoms N1 in the molecule located at

188

(x, y, z) act as hydrogen bond donor via H1N, to atoms O2 at (x-1, y, z).

189

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191

Figure 4: Crystal Structure of II

192 193

194

Figure 5: Packing Diagram of II

195 196 197

Table 1: Crystal Data and Structure Refinement of Compound I and II Crystal Data and Structure

I

Refinement Empirical formula Formula weight

C16H23NO4S 325.41

II C17H25NO4S 339.44

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Temperature/K

296(2)

Crystal system

Monoclinic

Space group

P21/n

296.15 orthorhombic P212121

a/Å

7.9687(12)

5.3382(3)

b/Å

25.817(4)

9.5791(9)

c/Å

8.2971(10)

36.078(3)

α/°

90

89.941(7)

β/°

102.959(13)

89.993(6)

γ/°

90

89.995(6)

1663.5(4)

1844.8(3)

Volume/Å3 Z

4

4

ρcalcmg/mm3

1.299

1.222

μ/mm-1

0.212

0.194

F(000)

696.0

728.0

Crystal size/mm3 2θ range for data collection Index ranges Reflections collected Independent reflections Data/restraints/parameters Goodness-of-fit on F2 Final R indexes [I>=2σ (I)]

0.27 × 0.08 × 0.04 5.946 to 57.958° -8 ≤ h ≤ 10, -24 ≤ k ≤ 35, -10 ≤ l ≤ 11 8267 3903[R(int) = 0.0558] 3903/1/202 0.984 R1 = 0.0694, wR2 =

0.420 × 0.270 × 0.210 6.208 to 59.014° -5 ≤ h ≤ 7, -13 ≤ k ≤ 9, 45 ≤ l ≤ 48 9718 4390[R(int) = 0.0472] 4390/0/213 1.019 R1 = 0.0634, wR2 = 10 | P a g e

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0.1351 Final R indexes [all data]

0.1249

R1 = 0.1491, wR2 =

R1 = 0.1492, wR2 =

0.1770

Largest diff. peak/hole / e Å-3

0.1647 0.22/-0.23

0.18/-0.26

Flack parameter

0.77(18)

198

Table 2: Hydrogen Bonds for Compound I

199

D

A

d(D-H)/Å

C12

H12 O11

0.93

2.58

3.447(4)

156.2

N1

H1N O41

0.858(10)

2.098(19)

2.878(4)

151(3)

1+X,

H

d(H-A)/Å

d(D-A)/Å

D-H-A/°

+Y, -1+Z

200

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Table 3: Bond Lengths of synthesized compounds

202

I Atoms

II

Experimental

Theoretical

Atoms

Length/Å

Experimental

Theoretical

Length/Å 1.539

S1

O1

1.430(4)

1.464

1.525(4)

1.537

S1

O2

1.442(4)

1.464

C7

1.526(4)

1.535

S1

N1

1.617(4)

1.681

C2

C3

1.521(4)

1.535

S1

C1

1.766(6)

1.798

C3

C4

1.538(4)

1.535

O3

C14

1.192(6)

1.213

C4

C5

1.513(5)

1.548

O4

C14

1.323(6)

1.354

C4

C15

1.500(4)

1.518

O4

C15

1.467(7)

1.449

C5

C6

1.524(4)

1.534

N1

C7

1.473(6)

1.468

C7

N1

1.467(4)

1.474

C1

C2

1.373(8)

1.395

C8

C9

1.384(5)

1.397

C1

C6

1.393(7)

1.397

C8

C13

1.388(4)

1.395

C2

C3

1.398(9)

1.394

C8

S1

1.763(3)

1.797

C3

C4

1.367(9)

1.401

C9

C10

1.371(5)

1.391

C4

C5

1.374(9)

1.403

C10 C11

1.392(5)

1.403

C4

C17

1.517(9)

1.509

C11 C12

1.379(5)

1.400

C5

C6

1.379(8)

1.392

C11 C14

1.515(5)

1.509

C7

C8

1.523(6)

1.535

C12 C13

1.387(4)

1.394

C8

C9

1.516(6)

1.540

C15 O3

1.329(4)

1.355

C8

C13

1.528(6)

1.540

C15 O4

1.205(4)

1.212

C9

C10

1.524(6)

1.533

C16 O3

1.450(4)

1.436

C10

C11

1.520(6)

1.534

N1

S1

1.593(3)

1.678

C11

C12

1.532(7)

1.547

O1

S1

1.430(2)

1.464

C11

C14

1.507(6)

1.520

C1

C2

C1

C6

C1

1.517(5)

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O2

S1

1.464

1.438(3)

C12

C13

1.528(6)

1.534

C15

C16

1.347(13)

1.520

203

Table 4: Bond Angles of synthesized compounds

204

I Atom

II

Experimental Theoretical

Experimental Theoretical

Atom

Angle/˚

Angle/˚

C2 C1 C6

109.8(3)

110.36

O1

S1

O2

120.6(3)

122.81

C2 C1 C7

112.1(3)

112.15

O1

S1

N1

106.4(3)

106.43

C6 C1 C7

112.1(3)

110.40

O1

S1

C1

108.3(2)

107.19

C1 C2 C3

111.9(3)

111.87

O2

S1

N1

106.9(2)

105.0

C2 C3 C4

111.4(3)

111.27

O2

S1

C1

106.4(3)

107.30

C5 C4 C3

110.5(3)

110.94

N1

S1

C1

107.8(3)

107.25

C15 C4 C3

108.4(3)

110.53

C14 O4

C15

118.8(6)

116.59

C15 C4 C5

112.7(3)

111.32

C7

N1

S1

119.7(4)

119.62

C4 C5 C6

112.6(3)

111.53

C2

C1

S1

121.3(4)

119.45

C5 C6 C1

111.2(3)

112.0

C2

C1

C6

119.1(6)

120.22

N1 C7 C1

111.5(3)

113.30

C6

C1

S1

119.6(5)

119.81

C9 C8 C13

119.8(3)

120.71

C1

C2

C3

119.9(6)

119.26

C9 C8 S1

120.6(2)

119.48

C4

C3

C2

121.6(7)

121.18

C10 C11 C14

121.2(4)

120.62

C3

C4

C5

117.8(7)

118.39

C12 C11 C10

117.9(3)

118.39

C3

C4

C17

120.8(7)

120.69

C12 C11 C14

121.0(3)

120.95

C5

C4

C17

121.4(6)

120.89

C11 C12 C13

121.7(3)

121.20

C4

C5

C6

122.2(6)

121.20

C12 C13 C8

119.2(4)

119.2

C5

C6

C1

119.5(6)

119.23

O3 C15 C4

112.6(3)

111.01

N1

C7

C8

112.4(4)

111.57

O4 C15 C4

125.6(4)

125.92

C7

C8

C13

109.6(4)

109.82 13 | P a g e

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O4 C15 O3

121.7(3)

123.05

C9

C8

C7

112.3(4)

112.87

C7 N1 S1

123.7(3)

121.8

C9

C8

C13

111.1(4)

110.14

C15 O3 C16

117.2(3)

115.30

C8

C9

C10

111.7(4)

112.33

N1 S1 C8

108.14(15)

106.89

C11 C10

C9

111.9(4)

111.56

O1 S1 C8

108.02(17)

107.50

C10 C11 C12

109.6(4)

110.74

O1 S1 N1

106.27(14)

104.86

C14 C11 C10

112.2(4)

111.41

C14 C11 C12

110.6(4)

110.27

C13 C12 C11

110.7(4)

111.29

C8 C13 C12

112.2(4)

112.33

O3 C14

O4

122.9(5)

123.79

O3 C14 C11

126.9(5)

125.38

O4 C14 C11

110.2(5)

110.81

C16 C15

112.4(9)

111.34

O4

205 206

3.1

Biological Studies

207

Anti-oxidants serve to protect our body from harms of reactive oxygen species (ROS) like

208

hydrogen peroxide, hydroxyl ion, superoxide and hydroxyl free radical. These oxidants are

209

harmful for our body e.g. converting the enzymes from their active form to non-active one and

210

rupturing of DNA strand [42]. ROS are involved in aging and cell death [43]. In our research

211

work, anti-oxidant potential of carboxylate esters derived from sulfonamide was evaluated by

212

using DPPH scavenging method. Compound (I) (65.27 ± 0.8%: IC50; 173.42 ± 1.2 µg) and (II)

213

(71.05 ± 1.2 %: IC50; 141.18 ± 0.7 µg) showed good anti-oxidant potential which is comparable

214

to gallic acid (77.54 ± 1.4%: IC50; 14.23 ± 0.4µg) a standard anti-oxidant molecule and

215

maximum activity was depicted by (II) as shown in Table 5.

216

Enzymes are the bio-catalyst which acts to control whole body functioning. Any disturbance in

217

an enzyme structure or their over production in body leads to harmful effects in body thus causes

218

major disease. In order to get rid of the diseases caused by enzyme’s extra activity in body, the 14 | P a g e

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219

particular enzyme must be denatured or its activity must be ceased by the use of enzyme

220

inhibiting agents. The synthesized compounds were screened for their inhibitory activity toward

221

enzymes. Acetylcholine esterase (AChE) is the enzyme that is involved in transmission of

222

neurotransmitter acetylcholine in brain and converts the acetylcholine into choline and acetate.

223

Over-functioning of AChE resulted in deficiency of acetylcholine in brain leading to memory

224

loss and hence causes Alzheimer’s disease. In order to cure disease caused by AChE, this

225

enzyme activity must be lowered down or inhibited [44]. Butyrylcholine esterase enzyme

226

(BChE) is also involved in conversion of acetylcholine into choline and acetate and hence the

227

resultant over-activity is related to Alzheimer’s disease [45]. In our research work, esters (I) and

228

(II) have been checked for their enzyme inhibition activity toward both of these enzymes.

229

Compound (I) showed 57.25 ± 1.3 and 53.08 ± 1.1 % inhibition toward AChE and BChE

230

respectively. Whereas (II) gave 62.72 ± 1.4 % inhibition toward AChE while 59.78 ± 0.8 %

231

inhibition toward BChE. α- Chymotrypsin (Protease) enzyme mainly functions in hydrolysis of

232

peptide linkage of proteins. Any over-activity of this enzyme is associated with various diseases

233

in body i.e. joints associated diseases, lung’s diseases and tumor growth [46,47]. α-

234

Chymotrypsin protease enzyme inhibition study was also part of our research work. Synthesized

235

compounds were applied for their inhibition effects against this enzyme. Results showed that

236

synthesized compounds (I) and (II) have 72.56 ± 0.9 and 69.47 ± 1.5% inhibition potential

237

respectively (Table 5).

238

In current research task, the synthesized compounds were also evaluated for their anti-bacterial

239

potential against nine bacterial strains. Both compounds were active against H. halophila, E. coli

240

and S. aureus and their activities were near to standard drugs. However, in remaining were

241

remained inactive and against both fungal strains, synthesized compounds exhibited no activity

242

as shown in Table 6.

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Table 5: Antioxidant and enzyme inhibition potential of I and II

244

Sr #

Sample

DPPH

Enzyme inhibition AChE

*%age

245

**IC

50

*%age

BChE

**IC

50

*%age

Protease

**IC

50

*%age

**IC

50

1

I

65.27 ± 0.8

173.42 ± 1.2

57.25 ± 1.3

183.27 ± 0.7

53.08 ± 1.1

196.13 ± 1.4

72.56 ± 0.9

102.72 ± 1.5

2

II

71.05 ± 1.2

141.18 ± 0.7

62.72 ± 1.4

125.49 ± 1.3

59.78 ± 0.8

178.08 ± 1.1

69.47 ± 1.5

137.53 ± 1.3

3

GA

77.54 ± 1.4

14.23 ± 0.4

-

-

-

-

-

-

4

PMSF

-

-

-

-

-

85.06 ± 0.7

* 100 µL(5 mg/mL)

-

** µg

37.41 ± 0.5

GA; Gallic acid PMSF; standard

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Table 6: Antimicrobial activity of I and II

247

Zone of Inhibition (mm)

Sample A

B

C

D

E

F

G

H

I

J

K

I

5.8 ± 0.8

NIL

NIL

NIL

12.5 ± 1.1

NIL

11.2 ± 0.7

13.7 ± 0.6

NIL

NIL

NIL

II

6.5 ± 1.0

NIL

NIL

NIL

NIL

NIL

15.7 ± 0.5

9.1 ± 0.8

NIL

NIL

NIL

Kanamycin

NIL

NIL

NIL

NIL

NIL

NIL

20.4 ± 0.9

19.3 ± 1.2

NIL

-

-

Ampicillin

26.4 ± 1.4

25.2 ± 1.0

31.5 ± 1.3

50.2 ± 1.7

45.5 ± 0.9

45.3 ± 1.2

26.1 ± 0.8

NIL

30.5 ± 1.1

-

-

Streptomycin

24.8 ± 1.7

24.1 ± 1.1

NIL

35.7 ± 1.3

15.6 ± 0.7

15.2 ± 1.0

25.8 ± 0.9

20.4 ± 1.1

25.7 ± 0.8

-

-

-

-

-

-

-

-

-

-

30.1 ± 1.5

28.4 ± 0.9

Fungizone

-

248 249 A; H. halophila: B; H. salina: C; S. sonnei: D; B. subtilis: E; C. salexigens: F; C. israelensis: G; E. coli: H; S. aureus: I; K. pneumoniae: J; A. niger: K; A. alternata 250 251

Density functional theory calculations have been performed not only to compare the theoretical

252

data with the experimental but also to gain insight into the packing and interaction energies of

253

molecules. The optimized geometries of compounds (I) and (II) are shown in Figure 6 whereas

254

the calculated geometric parameters are compared to the experimental ones in Table 3 and 4.

255

Compound (I) and (II) are structurally very similar, differing only in the ester part. Compound

256

(I) is a methoxy ester whereas compound (II) is an ethoxy ester. The rest of the structures are

257

identical therefore, their geometric parameters are also comparable. Both compounds contain a

258

sulfonamide and an ester functionality each. The calculated S=O bond length (for both S=O

259

bonds) is 1.464 Å for compounds (I) and (II) whereas the experimental S=O bond lengths differ

260

not only between the molecules but also for two S=O bonds in a molecule. The experimental

261

S=O bond lengths in compound (I) are 1.430 and 1.438 Å whereas the similar bond lengths in

262

compound (II) are 1.430 and 1.442 Å. The experimental and calculated C-S bond lengths in

263

compound (I) are 1.763 and 1.797 Å, respectively whereas the corresponding bond lengths in

264

compound (II) are 1.766 and 1.798 A, respectively. The calculated geometric parameters, in

265

general, show good agreement with the experimental geometric parameters. However, the 17 | P a g e

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266

maximum deviation in the bond lengths is observed for S-N bond lengths where the calculated

267

value for compound (I) (1.678 Å) is somewhat overestimated than the experimental 1.593 Å.

268

The calculated geometric parameters of the ester moiety also agree with the experimental ones.

269

For example, the calculated C=O bond length is 1.212 Å compared to 1.205 Å for the X-ray

270

structure. The C=O bond length of compound (II) shows even better corroboration with the

271

experiment (compare 1.192 and 1.213 for experimental and theoretical, respectively). The

272

efficiency of the theoretical methods for geometric parameters is presented here through root

273

mean square deviations (RMSD). The RMSD for bond lengths for compound (I) and (II) are

274

0.025 and 0.042.

275

276 277

Figure 6: Comparison of the optimized (right) and X-ray geometries (left) of compound 1

278

(top) and 2 (bottom)

279

Bond angles are also compared between the calculated and the X-ray structure. The B3LYP

280

functional performed quite good here as well. Most of the calculated bond angles were within 1

281

degree to the experimental values except a few where the deviation exceeds more than a degree.

282

The maximum deviation of 2.13 degrees in bond angle in compound (I) is observed for C15-C4-

283

C3, an angle which describes the orientation of the ester moiety with respect to the benzene ring.

284

For compound (II), a couple of bond angles deviate more than two degrees; O1-S1-O2 and C14-

285

O4-C15. The former bond angle is present within a sulfonamide group whereas the latter angle

286

represents the orientation of the ethoxy group with respect to carbonyl.

287

3.2

Packing behavior 18 | P a g e

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288

Next, the packing of both compounds in X-ray crystal was considered. Theoretical calculations

289

have been performed for the dimers of compound (I) and (II) and the interaction energies are

290

calculated. For this purpose, input structures were taken from the X-ray geometry. Analysis of

291

the results in Figure 7 reveals that the interaction sites are different for both compounds. In

292

compound (I), the sulfonamide group is in interaction with the protons of cyclohexane moiety.

293

On the other hand, in compound (II), the sulfonamide interacts with the hydrogen atoms of the

294

ethoxy chain. The difference in the interaction is also reflected in the interaction energies.

295

The calculated interaction energies between two molecules in (I) and (II) are 3.18 and 5.30 kcal

296

mol-1. For the packing of third molecule, compound (II) has different behavior than that of

297

compound (I). For compound (I), the nature of interaction is the same as that of dimer. But, for

298

compound (II), the third molecule orients itself in a different way (Figure 8) where aromatic ring

299

interacts with the sulfonamide group. The strength of this additional interaction is estimated

300

about 2.69 kcal mol-1.

301 302

Figure 7: Illustration of the optimized geometries of dimeric (I) and (II), calculated at

303

B3LYP/6-31G(d,p)

304

19 | P a g e

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305 306

Figure 8: Illustration of the packing of compound (II) in the optimized geometry, calculated at

307

B3LYP/6-31G(d,p)

308

309

HOMO

310

LUMO

311

Figure 9. Illustration of HOMOs and LUMOs of compounds (I) (top) and (II) (bottom),

312

calculated at B3LYP/6-31G(d,p)

313

3.3

314

The orbitals of compounds (I) and (II) are also analyzed to gain insight into the distribution of

315

densities in the frontier molecular orbitals (Figure 9). Both compounds show similar distribution

316

of densities in their HOMOs and LUMOs. The HOMOs are mainly localized on the nitrogen

317

atoms of the sulfonamide with some density on the aromatic ring of the sulfonamide whereas the

HOMO-LUMO Study

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318

LUMOs are mainly centered on the aromatic ring. The HOMO-LUMO gaps for compounds (I)

319

and (II) are 5.96 and 5.97 eV. The HOMO-LUMO gaps for both compounds are quite high

320

which indicate their kinetic stability. Since both structures differe only in the aliphatic chain

321

therefore, any significant different in the HOMO-LUMO gaps are not expected.

322

Conclusion

323

Sulfonamide’s carboxylate esters (I) and (II) were synthesized successfully by reacting alcohol

324

(ethanol and methanol) and sulfonamide ligand in acid catalyzed medium. Structures of

325

synthesized compounds were elucidated by using FTIR and single crystal X-ray crystallography.

326

Furthermore, density functional theory (DFT) was done through Gaussian software which also

327

supported the experimental crystallographic data. Both (I) and (II) were screened for their

328

biological potential as enzyme inhibition, antioxidant, antibacterial and antifungal agents. (I) and

329

(II) exhibited appreciable antioxidant and enzyme inhibition potential. They also showed

330

moderate behavior toward anti-bacterial and no activity against fungal strains.

331 332 333 334

ACKNOWLEDGMENT

335

The help of Higher Education Commission is acknowledged for funding this study under the

336

Project No. 20-2549/NRPU/R&D/HEC/12.

337

References

338

[1]

J.E. Toth, G.B. Grindey, W.J. Ehlhardt, J.E. Ray, G.B. Boder, J.R. Bewley,

339

Klingerman, S.B.

340

Sulfonimidamide Analogs of Oncolytic Sulfonylureas, J. Med. Chem. 40 (1997) 1018-

341

1025.

342

[2]

Gates, S.M.

Rinzel, R.M.

Schultz, L.C.

K.K.

Weir, J.F. Worzalla,

H.Yoshino, N.Ueda, J. Niijima, H. Sugumi, Y. Kotake, N. Koyanagi, K. Yoshimatsu,

343

M. Asada, T. Watanabe, Novel sulfonamides as potential, systemically active antitumor

344

agents, J. Med. Chem. 35 (1992) 2496-2497.

21 | P a g e

ACCEPTED MANUSCRIPT

345

[3]

T. Owa, H. Yoshino, T. Okauchi, K. Yoshimatsu, Y. Ozawa, N.H. Sugi, T. Nagasu, N.

346

Koyanagi, K. Kitoh, Discovery of novel antitumor sulfonamides targeting G1 phase of the

347

cell cycle, J. Med. Chem. 42 (1999) 3789-3799.

348

[4]

some pyrazole-sulfonamide derivatives. Med. Chem. Res. 23 (2014) 1278-1289.

349 350

S. Mert, S.Y. Ayse, I. Demirtas, K. Rahmi, Synthesis and antiproliferative activities of

[5]

S.H. Abdel-Hafez,

M.A. Hussein, Selenium-Containing Heterocycles: Synthesis and

351

Pharmacological Activities of Some New 4-Methylquinoline-2(1H) Selenone Derivatives,

352

Arch. Pharm. Chem. Life. Sci. 341 (2008) 240 - 246.

353

[6]

237 (1962) 536-540.

354 355

[7]

G. Mustafa, I.U. Khan, M. Ashraf, I. Afzal, S.A. Shahzad, M. Shafiq, Synthesis of new sulfonamides as lipoxygenase inhibitors, Bioorg. Med. Chem. 20 (2012) 2535-2539.

356 357

G. M. Brown, The Biosynthesis of Folic Acid: ii. Inhibition by sulfonamides, J. Bio. Chem.

[8]

C.P. Kordik, A.B. Reitz, Synthesis and Evaluation of a Series of Novel 2-[(4-

358

Chlorophenoxy)methyl]- benzimidazoles as Selective Neuropeptide Y Y1 Receptor

359

Antagonists, J. Med. Chem. 42 (1999) 181-201.

360

[9]

L.J. Beeley, J. M. Berge, H. Chapman, D.K. Dean, J. Kelly, K. Lowden, N.R. Kotecha,

361

H.K.A. Morgan, H.K. Rami, M. Thompson, A.K.K. Vong, R.W. Ward, A simplified

362

template approach towards the synthesis of a potent beta-3 adrenoceptor agonist at the

363

human receptor, Bioorg. Med. Chem. Lett. 7 (1997) 219-224.

364

[10] B. Hu, J. Ellingboe, S. Han, E. Largis, R. Mulvey, A. Oliphant, F.W. Sum, J. Tillett, (4-

365

Piperidin-1-yl)phenyl Amides:  Potent and Selective Human β3 Agonists, J. Med. Chem.

366

44 (2001) 1456-1466.

367

[11] M. Thakur, A. Thakur, P.V. Khadikar, C.T. Supuran, QSAR study on pKa vis-à-vis

368

physiological activity of sulfonamides: a dominating role of surface tension (inverse steric

369

parameter), Bioorg. Med. Chem. Lett. 15 (2005) 203-209.

370 371

[12] A. Kołaczek, I. Fusiarz,

J. Ławecka,

D. Branowsk, Biological activity and synthesis of

sulfonamide derivatives: a brief review, CHEMIK, 68 (2014) 620-628.

372

[13] H.M. Guo, L.T. Wang , J. Zhang , P.S. Zhao, F,F, Jian, Synthesis, IR Spectra, Crystal

373

Structure and DFT Studies on 1-Acetyl-3-(4-Chlorophenyl)-5-(4-Methylphenyl)-2-

374

Pyrazoline, Molecules 13 (2008) 2039-2048.

22 | P a g e

ACCEPTED MANUSCRIPT

375 376 377 378 379 380 381 382

[14] M.J. Frisch, G.W. Trucks, H.B. Schlegel, G.E. Scuseria, M.A. Robb, J.R. Cheeseman, G. Scalmani, V. Barone, B. Mennucci, G.A. Petersson, Gaussian 09, Wallingford, CT, 2009. [15] A.D. Becke, Density functional thermochemistry. III. The role of exact exchange, J. Chem. Phys. 98 (1993) 5648-5652. [16] C. Lee, W. R. Yang, G. Parr, Development of the Colle-Salvetti correlation-energy formula into a functional of the electron density, Phys. Rev. B: 37 (1988) 785-789. [17] P.C. Hariharan, J.A. Pople, The influence of polarization functions on molecular orbital hydrogenation energies, Theor. Chim. Acta, 28 (1973) 213-222.

383

[18] A. Ilyas, N. Muhammad, M.A. Gilani, K. Ayub, I.F.J. Vankelecom, A.L. Khan, Supported

384

protic ionic liquid membrane based on 3-(trimethoxysilyl) propan-1-aminium acetate for

385

the highly selective separation of CO2, J. Memb. Sci. 543 (2017) 301-309.

386

[19] S. Sherzaman, S.U. Rehman, M.N. Ahmed, B.A. Khan, T. Mahmood, K. Ayub, M. N.

387

Tahir, Thiobiuret based Ni(II) and Co(III) complexes: Synthesis, molecular structures and

388

DFT studies, J. Mol. Struct. 1148 (2017) 388-396.

389

[20] M. Nadeem,

U. Yunus, M.H. Bhatti,

K. Ayub,

M. Mehmood,

M.J. Saif,

390

Crystal structure, spectroscopic, electronic, luminescent and nonlinear optical properties of

391

(S)-4-Amino-5-(1-hydroxy-ethyl)-2,4-dihydro-[1,2,4]triazole-3-thione:

392

experimental and DFT study, J. Phys. Chem. Solids. 110 (2017) 218-226.

393 394

A

combined

[21] R.U. Nisa, K. Ayub, Mechanism of Zn(OTf)2 catalyzed hydroamination–hydrogenation of alkynes with amines: insight from theory, New J. Chem. 41 (2017) 5082-5090.

395

[22] T.U. Rahman, G. Uddin, R.U. Nisa, R. Ludwig, W. Liaqat, T. Mahmood, G. Mohammad,

396

M.I. Choudhary, K. Ayub, Spectroscopic and density functional theory studies of 7-

397

hydroxy-3'-methoxyisoflavone: A new isoflavone from the seeds of Indigofera heterantha

398

(Wall), Acta Part A Mol. Biomol. Spectrosc. 148 (2015) 375-381.

399

[23] M.N. Arshad, A.M. Asiri, K.A. Alamry, T. Mahmood, M.A. Gilani, K. Ayub, A.S. Birinji,

400

Synthesis, crystal structure, spectroscopic and density functional theory (DFT) study of N-

401

[3-anthracen-9-yl-1-(4-bromo-phenyl)-allylidene]-N-benzenesulfonohydrazine,

402

Spectrochim, Acta Part A Mol. Biomol. Spectrosc. 142 (2015) 364-374.

403 404

[24] F. Sattar, Z. Ullah, A. Rahman, A. Rauf, M.Tariq,

A.A. Tahir, K. Ayub, H. Ullah,

Phytochemical, spectroscopic and density functional theory study of Diospyrin, and non-

23 | P a g e

ACCEPTED MANUSCRIPT

405

bonding interactions of Diospyrin with atmospheric gases, Spectrochim. Acta Part A Mol.

406

Biomol. Spectrosc. 141 (2015) 71-79.

407

[25] T. Mahmood, N. Kosar, K. Ayub, DFT study of acceleration of electrocyclization in

408

photochromes under radical cationic conditions: Comparison with recent experimental

409

data, Tetrahed. 73(2017) 3521-3528.

410

[26] S. Bibi, H. Ullah, S.M. Ahmad, S.A.U.H. Ali, S. Bilal, A.A. Tahir, K. Ayub, Molecular

411

and electronic structure elucidation of polypyrrole gas sensors, J. Phys. Chem. C. 119

412

(2015) 15994-16003.

413

[27] S. Shahid, M.A. Raza, S.U.Rehman, Synthesis, characterization and antimicrobial potential

414

of transition metal complexes of triacetic lactone, Afr. J. Biotech. 8 (2009) 5116-5121.

415

[28] D. Shahwar, M.A. Raza, A.S.M. Mirza, M.A. Abbasi, V.U. Ahmed, Antioxidant potential

416 417

of phenolic extracts of Mimusops elengi, J. Chem. Soc. Pak. 32 (2010) 357-362. [29] G.L. Ellman,

K.D. Courtney,

V.Andres,

R.M. Feather-stone, A new and rapid

418

colorimetric determination of acetylcholinesterase activity, Biochem. Pharmocol. 7 (1961)

419

88-95.

420 421

[30] M. A. Raza, D.Shahwar, V.U. Ahmed, Protease inhibition activity of artemisinin and its biotransformed product, J. Chem. Soc. Pak. 35 (2013) 135-138.

422 423 424

[31] E.C. Jensen, C. Ogg, K. Nickerson, Lipoxygenase inhibitors shift the yeast/mycelium dimorphism in Ceratocystisulmi. Appl. Environ. Microb. 58 (1992) 2505-2508.

425

[32] C. Kemal, P. Louis-Flemberg, R. Krupinski-Olsen, A.L. Shorter, Reductive inactivation of

426

soybean lipoxygenase 1 by catechols: a possible mechanism for regulation of lipoxygenase

427

activity, J. Biochem. 26 (1987) 7064-7072.

428

[33] Agilent CrysAlis PRO Agilent Technologies, Yarnton, England (2012)

429

[34] G.M. Sheldrick, A short history of SHELXL, Acta Crystallogr. Sect. A, 64 (2008) 112-122.

430

[35] J.L. Farrugia, WinGX and ORTEP for Windows: an update, J. Appl. Cryst. 45 (2012) 849-

431 432 433

854. [36] A.L. Spek, Structure validation in chemical crystallography, Acta Crystallogr. Sect. D. 65 (2009), 148-155.

434 435

[37] J.L. Farrugia, WinGX Program Features, J. App. Cryst. 32 (1999) 837-838. 24 | P a g e

ACCEPTED MANUSCRIPT

436

[38] M. Ashfaq, S. Iram, M. Akkurt, I.U. Khan, G. Mustafa, S. Sharif, 4-[(4-Methyl-benzene-

437

sulfonamido)-meth-yl]cyclo-hexa-necarb-oxy-lic acid, Acta Crystallogr. Sect. A. 67

438

(2011), 01563.

439

[39] M. Ashfaq, N.M. Arshad, M. Danish M.A. Asiri, S. Khatoon, G. Mustafa, N.P.

440

Zolotarev, A.R. Butt, O. Şahin, Synthesis and description of intermolecular interactions in

441

new sulfonamide derivatives of tranexamic acid, J. Mol. Struct. 1103 (2016) 271-280.

442

[40] D. Cremer, A.J. Pople, General definition of ring puckering coordinates, J. Am. Chem. Soc. 97 (1975) 1354-1358.

443 444

[41] J. Bernstein,

E.R. Davis, L. Shimoni, N.L. Chang, Patterns in Hydrogen Bonding:

445

Functionality and Graph Set Analysis in Crystals, Angew. Chem. Int. Ed. Engl. 34 (1995)

446

1555-1573.

447

[42] V.O. Dolomanov, J.L. Bourhis, J.R. Gildea, K.A. J. Howard,

Puschmann, H. OLEX2: a

448

complete structure solution, refinement and analysis program, J. Appl. Cryst. 42 (2009)

449

339-341.

450

[43] J.R. Soares, T.C. Dinis, A.P. Cunha, L.M. Almeida, Antioxidant activities of some extracts of Thymus zygis. Rad. Res. 26 (1997) 469-478.

451 452

[44] L. Stefanis, R.E. Burke, L.A. Greene, Apoptosis in neurodegenerative disorders, Curr. Opin. Neurol. 10 (1997) 299-305.

453 454

[45] P.T. Francis, A.M. Palmer, M. Snape, K.G. Wilcock, The cholinergic hypothesis of

455

Alzheimer's disease: a review of progress, J. Neurology Neurosurgery

456

(1999) 137-147.

Psychiatry, 66

457

[46] A. Giovanni, P.F. David, Protease inhibitors in the clinic. Med. Chem. (2005) 71-104.

458

[47] J. Kaur, M.Q. Zhang, Molecular modelling and QSAR of reversible acetylcholines-terase inhibitors, Curr. Med. Chem. 7 (2000) 273-294.

459 460

1.

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Highlights  Two ester were synthesized and characterized through XRD analysis  DFT studies used to compare experimental and theoretical parameters  In-Vitro antimicrobial, antioxidant and enzyme inhibition potential were checked in order to explore their biological importance.