N. Argüelles et al. / Bioorg. Med. Chem. 18 (2010) 4238–4248
4247
compounds for 30 min at 37 °C, followed by the standard enzyme
assay. The synthetic compounds were prepared at 1, 2, 3, 4 and
5 mM in DMSO. A DMSO control was tested to exclude the inhibi-
tion of this solvent on the activity of the enzyme. After measuring
HMGR inhibition, mean IC50 values were compared to each other
and evaluated by the Tukey post hoc test with the two-way ANO-
VA. Significant differences were determined with a value of
P <0.05. Statistical analysis was carried out using the SigmaStat
software.
L.G.-S., M.C.C., G.C.-C., and J.T. are fellows of the EDI/IPN and COF-
AA/IPN programs.
References and notes
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A previously validated docking protocol we successfully used
to explain the binding mode of
a
-asarone (1) with HMGR20 was
also employed in this work. Thus, automated docking with the
program AUTODOCK, version 3.0.44 was employed to locate the
appropriate binding orientation and conformation of compounds
1, 11a–b, 17, 18, and 20a–d with HMGR. Docking was performed
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prepared with the program Molecular Operating Environment
(MOE), version 2008.10,46 and visualizations were carried out
with Maestro, version 9.0.47 All water and adenosine-50-diphos-
phate molecules were removed from the original Protein Data
Bank file. Polar hydrogen atoms were added and Kollman
charges,48 atomic solvation parameters and fragmental volumes
were assigned to the protein using AutoDock Tools (ADT). For
docking calculations, Gasteiger-Marsili partial charges49 were as-
signed to the ligands and non-polar hydrogen atoms were
merged. All torsions were allowed to rotate during docking. The
auxiliary program AutoGrid generated the grid maps. Each grid
was centered at the crystal structure of rosuvastatin. The grid
dimensions were 23 ꢁ 23 ꢁ 23 Å3 with points separated by
0.375 Å. Lennard-Jones parameters 12–10 and 12–6, supplied
with the program, were used for modeling hydrogen bonds and
van der Waals interactions, respectively. The distance-dependent
dielectric permittivity of Mehler and Solmajer50 was used for the
calculation of the electrostatic grid maps. For all ligands, random
starting positions, random orientations, and torsions were used.
The translation, quaternion, and torsion steps were taken from
default values in AutoDock. The Lamarckian genetic algorithm
and the pseudo-Solis and Wets methods were applied for minimi-
zation, using default parameters. The number of docking runs was
100. The complexes of the ligands with HMGR resulting from
molecular docking were further structurally optimized with the
MMFF94x force field implemented in MOE until the gradient
0.001 was reached. The default parameters implemented into
the MOE’s LigX46 application were used as we have previously de-
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We thank Bruce Allan Larsen for reviewing the use of English in
this manuscript and to Dr. Narender Singh for helpful discussions.
We are very grateful to Dr. Fabián López-Vallejo for critically read-
ing the manuscript and his valuable comments. We thank CONA-
CyT for financing projects G-34851-M and Salud-69984, and
DGAPA for project NI 202101. N.A., E.S.-S., A.M., and F.J. are grateful
to CONACyT and PIFI-IPN for the scholarships awarded. J.L.M.-F. is
grateful to the State of Florida, Executive Office of the Governor’s
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