Design, Synthesis and biological evaluation of
novel triazole, urea and thiourea derivatives of
Quinoline against Mycobacterium tuberculosis
Ram Shankar Upadhayaya,a Girish M. Kulkarni,a Nageswara Rao Vasireddy,a Jaya
Kishore Vandavasi,a Shailesh S. Dixit,a Vivek Sharma,a Jyoti Chattopadhyaya*,b
aInstitute
of Molecular Medicine, Pune 411 057, India
b
Program of Bioorganic Chemistry, Institute of Cell and Molecular Biology, Biomedical
Centre, Uppsala University, Sweden
S1
Table T1:
Sr. No.
Contents
Page No.
1.
1
H-NMR spectra of compound 2
S8
2.
D2O-NMR spectra of compound 2
S9
3.
13
C-NMR spectra of compound 2
S10
4.
DEPT-NMR spectra of compound 2
S11
5.
1
S12
6.
13
C-NMR spectra of compound 3
S13
7.
DEPT-NMR spectra of compound 3
S14
8.
1
S15
9.
13
C-NMR spectra of compound 4
S16
10.
DEPT-NMR spectra of compound 4
S17
11.
1
H-NMR spectra of compound 5
S18
12.
D2O-NMR spectra of compound 5
S19
13.
13
C-NMR spectra of compound 5
S20
14.
DEPT-NMR spectra of compound 5
S21
15.
1
S22
17.
13
C-NMR spectra of compound 6
S23
18.
DEPT-NMR spectra of compound 6
S24
19.
1
S25
20.
13
C-NMR spectra of compound 7
S26
21.
DEPT-NMR spectra of compound 7
S27
22.
1
H-NMR spectra of compound 8
S28
23.
D2O-NMR spectra of compound 8
S29
24.
13
S30
H-NMR spectra of compound 3
H-NMR spectra of compound 4
H-NMR spectra of compound 6
H-NMR spectra of compound 7
C-NMR spectra of compound 8
S2
25.
DEPT-NMR spectra of compound 8
S31
26.
1
S32
27.
13
C-NMR spectra of compound 9
S33
28.
DEPT-NMR spectra of compound 9
S34
29.
1
S35
30.
13
C-NMR spectra of compound 10
S36
31.
DEPT-NMR spectra of compound 10
S37
32.
1
S38
33.
13
C-NMR spectra of compound 11
S39
34.
DEPT-NMR spectra of compound 11
S40
35.
1
S41
36.
13
C-NMR spectra of compound 12
S42
37.
DEPT-NMR spectra of compound 12
S43
38.
1
S44
39.
13
C-NMR spectra of compound 13
S45
40.
DEPT-NMR spectra of compound 13
S46
41.
1
H-NMR spectra of compound 14
S47
42.
D2O-NMR spectra of compound 14
S48
43.
13
C-NMR spectra of compound 14
S49
44.
DEPT-NMR spectra of compound 14
S50
45.
1
H-NMR spectra of compound 15
S51
46.
D2O-NMR spectra of compound 15
S52
47.
13
C-NMR spectra of compound 15
S53
48.
DEPT-NMR spectra of compound 15
S54
49.
1
H-NMR spectra of compound 16
S55
50.
D2O-NMR spectra of compound 16
S56
51.
13
S57
H-NMR spectra of compound 9
H-NMR spectra of compound 10
H-NMR spectra of compound 11
H-NMR spectra of compound 12
H-NMR spectra of compound 13
C-NMR spectra of compound 16
S3
52.
DEPT-NMR spectra of compound 16
S58
53.
1
H-NMR spectra of compound 17
S59
54.
D2O-NMR spectra of compound 17
S60
55.
13
C-NMR spectra of compound 17
S61
56.
DEPT-NMR spectra of compound 17
S62
57.
1
H-NMR spectra of compound 18
S63
58.
D2O-NMR spectra of compound 18
S64
59.
13
C-NMR spectra of compound 18
S65
60.
DEPT-NMR spectra of compound 18
S66
61.
1
H-NMR spectra of compound 19
S67
62.
D2O-NMR spectra of compound 19
S68
63.
13
C-NMR spectra of compound 19
S69
64.
DEPT-NMR spectra of compound 19
S70
65.
1
H-NMR spectra of compound 20
S71
66
D2O-NMR spectra of compound 20
S72
67
13
C-NMR spectra of compound 20
S73
68
DEPT-NMR spectra of compound 20
S74
69
1
H-NMR spectra of compound 21
S75
70
D2O-NMR spectra of compound 21
S76
71
13
C-NMR spectra of compound 21
S77
72.
DEPT-NMR spectra of compound 21
S78
73.
1
H-NMR spectra of compound 22
S79
74.
D2O-NMR spectra of compound 22
S80
75.
13
C-NMR spectra of compound 22
S81
76.
DEPT-NMR spectra of compound 22
S82
77.
1
H-NMR spectra of compound 23
S83
78.
D2O-NMR spectra of compound 23
S84
S4
79.
13
C-NMR spectra of compound 23
S85
80.
DEPT-NMR spectra of compound 23
S86
81.
1
H-NMR spectra of compound 24
S87
82.
D2O-NMR spectra of compound 24
S88
83.
13
C-NMR spectra of compound 24
S89
84.
DEPT-NMR spectra of compound 24
S90
85.
1
H-NMR spectra of compound 25
S91
86.
D2O-NMR spectra of compound 25
S92
87.
13
C-NMR spectra of compound 25
S93
88.
DEPT-NMR spectra of compound 25
S94
89.
1
H-NMR spectra of compound 26
S95
90.
D2O-NMR spectra of compound 26
S96
91.
13
C-NMR spectra of compound 26
S97
92.
DEPT-NMR spectra of compound 26
S98
93.
1
H-NMR spectra of compound 27
S99
94.
D2O-NMR spectra of compound 27
S100
95.
13
C-NMR spectra of compound 27
S101
96.
DEPT-NMR spectra of compound 27
S102
97.
1
H-NMR spectra of compound 28
S103
98.
D2O-NMR spectra of compound 28
S104
99.
13
C-NMR spectra of compound 28
S105
100.
DEPT-NMR spectra of compound 28
S106
101.
1
H-NMR spectra of compound 29
S107
102.
D2O-NMR spectra of compound 29
S108
103.
13
C-NMR spectra of compound 29
S109
104.
DEPT-NMR spectra of compound 29
S110
105.
1
S111
H-NMR spectra of compound 30
S5
106.
13
C-NMR spectra of compound 30
S112
107.
DEPT-NMR spectra of compound 30
S113
108.
1
H-NMR spectra of compound 31
S114
109.
D2O-NMR spectra of compound 31
S115
110.
13
C-NMR spectra of compound 31
S116
111.
DEPT-NMR spectra of compound 31
S117
112.
1
H-NMR spectra of compound 32
S118
113.
D2O-NMR spectra of compound 32
S119
114.
13
C-NMR spectra of compound 32
S120
115.
DEPT-NMR spectra of compound 32
S121
116.
1
H-NMR spectra of compound 33
S122
117.
D2O-NMR spectra of compound 33
S123
118.
13
C-NMR spectra of compound 33
S124
119.
DEPT-NMR spectra of compound 33
S125
120.
1
H-NMR spectra of compound 34
S126
121.
D2O-NMR spectra of compound 34
S127
122.
13
C-NMR spectra of compound 34
S128
123.
DEPT-NMR spectra of compound 34
S129
124.
1
H-NMR spectra of compound 35
S130
125.
D2O-NMR spectra of compound 35
S131
126.
13
C-NMR spectra of compound 35
S132
127.
DEPT-NMR spectra of compound 35
S133
128.
1
H-NMR spectra of compound 36
S134
129.
D2O-NMR spectra of compound 36
S135
130.
13
C-NMR spectra of compound 36
S136
131.
DEPT-NMR spectra of compound 36
S137
132.
1
S138
H-NMR spectra of compound 37
S6
133.
D2O-NMR spectra of compound 37
S139
134.
13
C-NMR spectra of compound 37
S140
135.
DEPT-NMR spectra of compound 37
S141
136.
1
H-NMR spectra of compound 38
S142
137.
D2O-NMR spectra of compound 38
S143
138.
13
C-NMR spectra of compound 38
S144
139.
DEPT-NMR spectra of compound 38
S145
140.
Biological methods
S146
141.
Table S1: % Inhibition and Standard Deviation data.
S147
142.
Theoretical methods
S148
143.
Cytotoxicity assay
S149
144.
References
S150
S7
O2N
2
N
H
O
S8
O2N
2
N
H
O
S9
O2N
2
N
H
O
S10
O2N
2
N
H
O
S11
O2N
N
Cl
3
S12
O2N
N
Cl
3
S13
O2N
N
Cl
3
S14
O2N
N
O
O
4
F
S15
O2N
N
O
O
4
F
S16
O2N
N
O
O
4
F
S17
H2N
N
O
O
5
F
S18
H2N
N
O
O
5
F
S19
H2N
N
O
O
5
F
S20
H2N
N
O
O
5
F
S21
N3
N
O
O
6
F
S22
N3
N
O
O
6
F
S23
N3
N
O
O
6
F
S24
N N
N
N
O
O
7
F
S25
N N
N
N
O
O
7
F
S26
N N
N
N
O
O
7
F
S27
N N
N
N
O
OH
8
F
S28
N N
N
N
O
OH
8
F
S29
N N
N
N
O
OH
8
F
S30
N N
N
N
O
OH
8
F
S31
N N
N
N
O
Cl
9
F
S32
N N
N
N
O
Cl
9
F
S33
N N
N
N
O
Cl
9
F
S34
N N
N
N
N
O
N
10
F
S35
N N
N
N
N
O
N
10
F
S36
N N
N
N
N
O
N
10
F
S37
N N
N
N
O
N
11
F
S38
N N
N
N
O
N
11
F
S39
N N
N
N
O
N
11
F
S40
F
N N
N
N
N
O
N
12
F
S41
F
N N
N
N
N
O
N
12
F
S42
F
N N
N
N
N
O
N
12
F
S43
N N
N
N
N
N
O
N
13
F
S44
N N
N
N
N
N
O
N
13
F
S45
N N
N
N
N
N
O
N
13
F
S46
H
N
H
N
O
NO 2
N
O
O
14
F
S47
H
N
H
N
O
NO 2
N
O
O
14
F
S48
H
N
H
N
O
NO 2
N
O
O
14
F
S49
H
N
H
N
O
NO 2
N
O
O
14
F
S50
H
N
O
H
N
O
N
O
O
15
F
S51
H
N
O
H
N
O
N
O
O
15
F
S52
H
N
O
H
N
O
N
O
O
15
F
S53
H
N
O
H
N
O
N
O
O
15
F
S54
H
N
H
N
O
O
N
O
O
16
F
S55
H
N
H
N
O
O
N
O
O
16
F
S56
H
N
H
N
O
O
N
O
O
16
F
S57
H
N
H
N
O
O
N
O
O
16
F
S58
H
N
O
H
N
O
N
O
O
17
F
S59
H
N
O
H
N
O
N
O
O
17
F
S60
H
N
O
H
N
O
N
O
O
17
F
S61
H
N
O
H
N
O
N
O
O
17
F
S62
H
N
H
N
S
N
O
O
18
Cl
F
S63
H
N
H
N
S
N
O
O
18
Cl
F
S64
H
N
H
N
S
N
O
18
Cl
F
S65
O
H
N
H
N
S
N
O
O
18
Cl
F
S66
H
N
O
H
N
S
N
O
O
19
F
S67
H
N
O
H
N
S
N
O
O
19
F
S68
H
N
O
H
N
S
N
O
O
19
F
S69
H
N
O
H
N
S
N
O
O
19
F
S70
H
N
H
N
S
O
N
O
O
20
F
S71
H
N
H
N
S
O
N
O
O
20
F
S72
H
N
H
N
S
O
N
O
O
20
F
S73
H
N
H
N
S
O
N
O
O
20
F
S74
H
N
O
H
N
S
N
O
O
21
F
S75
H
N
O
H
N
S
N
O
O
21
F
S76
H
N
O
H
N
S
N
O
O
21
F
S77
H
N
O
H
N
S
N
O
O
21
F
S78
H
N
H
N
O
NO 2
N
O
OH
22
F
S79
H
N
H
N
O
NO 2
N
O
OH
22
F
S80
H
N
H
N
O
NO 2
N
O
OH
22
F
S81
H
N
H
N
O
NO 2
N
O
OH
22
F
S82
H
N
O
H
N
O
N
O
OH
23
F
S83
H
N
O
H
N
O
N
O
OH
23
F
S84
H
N
O
H
N
O
N
O
OH
23
F
S85
H
N
O
H
N
O
N
O
OH
23
F
S86
H
N
H
N
O
O
N
O
OH
24
F
S87
H
N
H
N
O
O
N
O
OH
24
F
S88
H
N
H
N
O
O
N
O
OH
24
F
S89
H
N
H
N
O
O
N
O
OH
24
F
S90
H
N
O
H
N
O
N
O
OH
25
F
S91
H
N
O
H
N
O
N
O
OH
25
F
S92
H
N
O
H
N
O
N
O
OH
25
F
S93
H
N
O
H
N
O
N
O
OH
25
F
S94
H
N
H
N
S
N
O
OH
26
Cl
F
S95
H
N
H
N
S
N
O
OH
26
Cl
F
S96
H
N
H
N
S
N
O
OH
26
Cl
F
S97
H
N
H
N
S
N
O
OH
26
Cl
F
S98
H
N
O
H
N
S
N
O
OH
27
F
S99
H
N
O
H
N
S
N
O
OH
27
F
S100
H
N
O
H
N
S
N
O
OH
27
F
S101
H
N
O
H
N
S
N
O
OH
27
F
S102
H
N
H
N
S
O
N
O
OH
28
F
S103
H
N
H
N
S
O
N
O
OH
28
F
S104
H
N
H
N
S
O
N
O
OH
28
F
S105
H
N
H
N
S
O
N
O
OH
28
F
S106
H
N
O
H
N
S
N
O
OH
29
F
S107
H
N
O
H
N
S
N
O
OH
29
F
S108
H
N
O
H
N
S
N
O
OH
29
F
S109
H
N
O
H
N
S
N
O
OH
29
F
S110
O2N
N
O
O
Br
30
F
S111
O2N
N
O
O
Br
30
F
S112
O2N
N
O
O
Br
30
F
S113
O2N
N
O
OH
O
N
31
F
S114
O2N
N
O
OH
O
N
31
F
S115
O2N
N
O
OH
O
N
31
F
S116
O2N
N
O
OH
O
N
31
F
S117
O2N
N
O
OH
N
32
F
S118
O2N
N
O
OH
N
32
F
S119
O2N
N
O
OH
N
32
F
S120
O2N
N
O
OH
N
32
F
S121
O2N
N
O
N
OH
N
33
F
S122
O2N
N
O
N
OH
N
33
F
S123
O2N
N
O
N
OH
N
33
F
S124
O2N
N
O
N
OH
N
33
F
S125
O2N
N
O
34
OH
N
N
F
S126
O2N
N
O
34
OH
N
N
F
S127
O2N
N
O
34
OH
N
N
F
S128
O2N
N
O
34
OH
N
N
F
S129
O
O2N
N
O
OH
N
N
35
F
S130
O
O2N
N
O
OH
N
N
35
F
S131
O
O2N
N
O
OH
N
N
35
F
S132
O
O2N
N
O
OH
N
N
35
F
S133
O
O
2N
N
O
O
H
N
N
36
F
S134
O
O2N
N
O
36
OH
N
N
F
S135
O
O2N
N
O
36
OH
N
N
F
S136
O
O 2N
N
O
OH
N
N
36
F
S137
F
F
F
O2N
N
O
37
OH
N
N
F
S138
F
F
F
O2N
N
O
37
OH
N
N
F
S139
F
F
F
O2N
N
O
37
OH
N
N
F
S140
F
F
F
O2N
N
O
37
OH
N
N
F
S141
Cl
O2N
N
O
38
OH
N
N
F
S142
Cl
O2N
N
O
38
OH
N
N
F
S143
Cl
O2N
N
O
38
OH
N
N
F
S144
Cl
O2N
N
O
38
OH
N
N
F
S145
Antimycobacterial activity:
All the compounds were screened for antimycobacterial activity against mycobacterium tuberculosis by BACTEC 460 radiometric methods.
The broth based BACTEC 460 TB system was used for the growth of Mycobacteria. In this growth system, Mycobacterium tuberculosis
H37RV was grown in the C14 labelled substrate in 7H12 medium, substrate is utilized by growing mycobacteria and 14CO2 is produced, which
is detected in the form of ‘Growth Index’ (GI), which reflects the rate and amount of growth in the medium vial. If an antituberculosis drug
will be added to the medium vial, it will suppress the growth of the bacteria which is detected by the decrease of the GI values as compared to
the control vial. The positive controls taken in the experiment were the standard therapeutic drugs presently used for the treatment of the
tuberculosis, Isoniazid and Rifampin.
The rate of increase in GI values or the change in GI over the previous day GI is called delta (∆) GI, is compared with that of control values.
If ∆GI of the drug-containing vial is equal to or greater than (≥) than that in control vial the organisms are considered resistant to the drug. On
the contrary if ∆GI of the drug-containing vial is less (<) than the control vial the organisms are considered susceptible.
On the basis of the values calculated for ∆G, out of all screened compounds, four compounds i.e. 10, 22 and 24 were found to act as active
antimycobacterial agents. Below given table depicts ∆G of the compounds tested.
S146
Table S1: % Inhibition and Standard Deviation data.
Days
1
2
3
4
5
6
7
8
9
10
11
GI
3
3
4
23
15
13
43
29
21
58
33
31
63
46
37
69
52
27
74
53
44
66
55
44
61
55
43
53
49
36
51
41
32
∆
GI
-2
-8
-4
9
16
31
8
18
43
6
21
41
9
21
40
9
18
32
8
16
36
8
17
33
8
15
29
9
15
30
-
-
1
0
1
3
3
4
12
10
14
15
16
22
17
20
26
18
23
28
22
28
37
25
35
42
30
39
48
33
48
53
34
51
57
41
64
68
7
13
11
95.9
93.6
93.2
2.06
17
18
16
30
18
23
14
14
18
13
13
17
11
12
16
14
16
18
14
14
20
13
14
19
-
-
-
-1
0
-1
20
26
18
40
46
34
61
70
52
93
101
80
152
152
134
220
232
219
345
344
346
466
446
450
605
618
547
617
719
720
813
818
796
196
99
76
99.7
99.6
98.0
0.53
-
N N
N
N
N
O
N
10
SD
GI
F
H
N
H
N
O
N
O2N
O
OH
SD
%
Inhibition
94.9
95.9
96.8
1.34
99.1
98.5
97.0
1.30
22
F
GI
H
N
H
N
O
N
O
O
OH
SD
24
F
Isoniazid
GI
Untreated Control
SD
GI
S147
Theoretical methods:
Homology model consisting of one subunit A and two subunit C of the ATP-synthase of M. tuberculosis was developed from
corresponding subunits of Escherichia coli ATP synthase.1 MODELLER2 was used for generating homology model and protocol used
is same as mentioned before.3 All acidic and basic residues were kept in their default protonation states i.e. arginine and lysine,
protonated and glutamic and aspartic acid, deprotonated, except Glu61, which was protonated. 1,3
The selected ligand molecules were sketched (irrespective of stereochemistry) and energy minimized for 2500 steps in
Ghemical.4 Subsequently a random conformational search was performed on each molecule with 2000 steps of conformational search
and 300 steps of energy minimization. Final lowest energy structure of each ligand molecule was converted to *.pdbqt file using
AutoDock Tools5 (http://mgltools.scripps.edu/) and used for automated docking in AutoDock 4.6 Same as discussed before,3 using
AutoDock Tools and AutoDock 4, the ligand was docked into a cavity located near Arg186 (subunit A) and Glu61 (subunit C) of M.
tuberculosis ATP-synthase. The grid dimensions used were 64Åx42Åx44Å. AutoDock parameters like population size, energy
evaluations and number of Lamarckian Genetic Algorithm runs were 250, 25000000 and 100, respectively. Initial coordinates (tran0
option in AutoDock) were set to the defined grid center. The docked protein-ligand complex with lowest binding energy was selected
for further analysis. Further we analyzed the intermolecular energies as calculated by AutoDock docking calculation as they give more
precise description of binding.
The docked complexes of certain compounds were subjected to forcefield-based refinement. The ligand coordinates were
extracted from *.pdbqt files of complexes generated by AutoDock. Hydrogens were added with Ghemical and converted to *.mol2
format using Openbabel. The antechamber7 module of AMBER8 was used for writing the necessary files (*.prepin and *.frcmod) for
ligand molecules. The AM1-BCC9 charge model was used for ligand molecules, which has been suggested to perform very well,
relative to other well known methods for partial charge calculations.10 The General Amber Force Field (GAFF)11 and ff038 were used
for ligand and protein, respectively. The complexes were minimized for 10000 steps using sander tool available in AMBER. The
interaction energy (van der Waals and Electrostatic) between ligand and protein were computed using NAMDEnergy tool available in
NAMD12 for the minimized complex. During analysis, hydrogen bond criteria used was Donor-H......Acceptor distance < 3Å and
Donor...H...Acceptor angle > 150º.
Visualization of molecules was done with the help of Visual Molecular Dynamics (VMD)13 and all simulations were run on
2.2GHz AMD Dual Core machine, running with Red Hat Enterprise Linux 5.
S148
Cytotoxicity assay:
Cellular viability in the presence and absence of test compounds was determined by MTT (3- (4,5-dimethylthiazol-2yl)-2,5-dimethyl
tetrazolium bromide; Sigma-Aldrich) microcultured tetrazolium assay.14,15 The cells (monocytic cell line, THP-1) were plated in flatbottomed 96-well plates (10,000 cells/100µL) and cultured in controlled atmosphere (5% CO2 at 37 °C). Cells were cultured in the
presence of medium along with DMSO (live controls). Different concentrations of compounds (1µg/100µl to 10µg/100µl) were added
to the cells. After 24h and 72h, stock MTT solution (5 mg/mL) was added to the culture. Cells were again kept in CO2 incubator for
4h. After 4h, 100µl isopropanol was added and mixed 5-6 times. The absorbance was read at 540 nm in a plate reader (Bio-Rad—
450). The results were represented as percentage cell viability (Table 3). All the experiments were carried out in triplicates and the
readings were the average mean of three readings.
S149
References
1. Rastogi, V.; Girvin, M. Nature 1999, 402, 263.
2. Sali, A.; Blundell, T. L. J. Mol. Biol. 1993, 234, 779.
3. Upadhayaya, R. S.; Jaya Kishore, V.; Nageswar Rao, V.; Sharma, V.; Dixit, S. S.; Chattopadhyaya, J. Bioorg. Med. Chem.
2009, 17, 2830.
4. Hassinen, T.; Peräkylä, M. J. Comput. Chem. 2001, 22, 1229.
http://www.uku.fi/~thassine/projects/ghemical/
5. Sanner, M. F. J. Mol. Graph. Mod. 1999, 17, 57.
6. Morris, G. M.; Goodsell, D. S.; Halliday, R.S.; Huey, R.; Hart, W. E.; Belew, R. K.; Olson, A. J. J. Comput. Chem. 1998, 19,
1639.
7. Wang, J.; Wang, W.; Kollman, P. A.; Case, D. A. J. Mol. Graph. Mod. 2006, 25, 247.
8. Duan, Y.; Wu, C.; Chowdhury, S.; Lee, M. C.; Xiong, G.; Zhang, W.; Yang, R.; Cieplak, P.; Luo, R.; Lee, T.; Caldwell, J.;
Wang, J.; Kollman, P. J. Comput. Chem. 2003, 24, 1999.
9. Jakalian, A.; Jack, D. B.; Bayly, C. I. J. Comput. Chem. 2002, 23, 1623.
10. Mobley, D. L.; Dumont, I.; Chodera, J. D.; Dill, K. A. J. Phys. Chem. 2007, 111, 2242.
11. Wang, J.; Wolf, R. M.; Caldwell, J. W.; Kollman, P. A.; Case, D. A. J. Comput. Chem. 2004, 25, 1157.
12. Kalé, L.; Skeel, R.; Bhandarkar, M.; Brunner, R.; Gursoy, A.; Krawetz, N.; Phillips, J.; Shinozaki, A.; Varadarajan, K.;
Schulten, K. J. Comput. Phys. 1999, 151, 283.
13. Humphrey, W.; Dalke, A.; Schulten, K. J. Mol. Graph. 1996, 14, 33.
14. Souza, M. C.; Siani, A. C.; Ramos, M. F. S.; Limas, O. M., Jr.; Henrique, M. G. M. O. Pharmazie, 2003, 58, 582.
S150
15. Carvalho, M. V.; Penido, C.; Siani, A. C.; Valente, L. M. M.; Henriques, M. G. M. O. Gmelin Inflammopharmacol. 2006, 14, 48.
S151