36 research outputs found
Quantum Mechanics/Molecular Mechanics Modeling of Covalent Addition between EGFR–Cysteine 797 and <i>N</i>‑(4-Anilinoquinazolin-6-yl) Acrylamide
Irreversible
epidermal growth factor receptor (EGFR) inhibitors
can circumvent resistance to first-generation ATP-competitive inhibitors
in the treatment of nonsmall-cell lung cancer. They covalently bind
a noncatalytic cysteine (Cys797) at the surface of EGFR active site
by an acrylamide warhead. Herein, we used a hybrid quantum mechanics/molecular
mechanics (QM/MM) potential in combination with umbrella sampling
in the path-collective variable space to investigate the mechanism
of alkylation of Cys797 by the prototypical covalent inhibitor <i>N</i>-(4-anilinoquinazolin-6-yl) acrylamide. Calculations show
that Cys797 reacts with the acrylamide group of the inhibitor through
a direct addition mechanism, with Asp800 acting as a general base/general
acid in distinct steps of the reaction. The obtained reaction free
energy is negative (Δ<i>A</i> = −12 kcal/mol)
consistent with the spontaneous and irreversible alkylation of Cys797
by <i>N</i>-(4-anilinoquinazolin-6-yl) acrylamide. Our calculations
identify desolvation of Cys797 thiolate anion as a key step of the
alkylation process, indicating that changes in the intrinsic reactivity
of the acrylamide would have only a minor impact on the inhibitor
potency
Structure-Based Virtual Screening of MT<sub>2</sub> Melatonin Receptor: Influence of Template Choice and Structural Refinement
Developing GPCR homology models for
structure-based virtual screening
requires the choice of a suitable template and refinement of binding
site residues. We explored this systematically for the MT<sub>2</sub> melatonin receptor, with the aim to build a receptor homology model
that is optimized for the enrichment of active melatoninergic ligands.
A set of 12 MT<sub>2</sub> melatonin receptor models was built using
different GPCR X-ray structural templates and submitted to a virtual
screening campaign on a set of compounds composed of 29 known melatonin
receptor ligands and 2560 drug-like decoys. To evaluate the effect
of including a priori information in receptor models, 12 representative
melatonin receptor ligands were placed into the MT<sub>2</sub> receptor
models in poses consistent with known mutagenesis data and with assessed
pharmacophore models. The receptor structures were then adapted to
the ligands by induced-fit docking. Most of the 144 ligand-adapted
MT<sub>2</sub> receptor models showed significant improvements in
screening enrichments compared to the unrefined homology models, with
some template/refinement combinations giving excellent enrichment
factors. The discriminating ability of the models was further tested
on the 29 active ligands plus a set of 21 inactive or low-affinity
compounds from the same chemical classes. Rotameric states of side
chains for some residues, presumed to be involved in the binding process,
were correlated with screening effectiveness, suggesting the existence
of specific receptor conformations able to recognize active compounds.
The top MT<sub>2</sub> receptor model was able to identify 24 of 29
active ligands among the first 2% of the screened database. This work
provides insights into the use of refined GPCR homology models for
virtual screening
MT<sub>1</sub> and MT<sub>2</sub> Melatonin Receptors: Ligands, Models, Oligomers, and Therapeutic Potential
Numerous physiological functions
of the pineal gland hormone melatonin
are mediated via activation of two G-protein-coupled receptors, MT<sub>1</sub> and MT<sub>2</sub>. The melatonergic drugs on the market,
ramelteon and agomelatine, as well as the most advanced drug candidates
under clinical evaluation, tasimelteon and TIK-301, are high-affinity
nonselective MT<sub>1</sub>/MT<sub>2</sub> agonists. A great number
of MT<sub>2</sub>-selective ligands and, more recently, several MT<sub>1</sub>-selective agents have been reported to date. Herein, we review
recent advances in the field focusing on high-affinity agonists and
antagonists and those displaying selectivity toward MT<sub>1</sub> and MT<sub>2</sub> receptors. Moreover, the existing models of MT<sub>1</sub> and MT<sub>2</sub> receptors as well as the current status
in the emerging field of melatonin receptor oligomerization are critically
discussed. In addition to the already existing indications, such as
insomnia, circadian sleep disorders, and depression, new potential
therapeutic applications of melatonergic ligands including cardiovascular
regulation, appetite control, tumor growth inhibition, and neurodegenerative
diseases are presented
Quantum Mechanics/Molecular Mechanics Modeling of Fatty Acid Amide Hydrolase Reactivation Distinguishes Substrate from Irreversible Covalent Inhibitors
Carbamate
and urea derivatives are important classes of fatty acid
amide hydrolase (FAAH) inhibitors that carbamoylate the active-site
nucleophile Ser241. In the present work, the reactivation mechanism
of carbamoylated FAAH is investigated by means of a quantum mechanics/molecular
mechanics (QM/MM) approach. The potential energy surfaces for decarbamoylation
of FAAH covalent adducts, derived from the <i>O</i>-aryl
carbamate URB597 and from the <i>N</i>-piperazinylurea JNJ1661610,
were calculated and compared to that for deacylation of FAAH acylated
by the substrate oleamide. Calculations show that a carbamic group
bound to Ser241 prevents efficient stabilization of transition states
of hydrolysis, leading to large increments in the activation barrier.
Moreover, the energy barrier for the piperazine carboxylate was significantly
lower than that for the cyclohexyl carbamate derived from URB597.
This is consistent with experimental data showing slowly reversible
FAAH inhibition for the <i>N</i>-piperazinylurea inhibitor
and irreversible inhibition for URB597
Metadynamics Simulations Distinguish Short- and Long-Residence-Time Inhibitors of Cyclin-Dependent Kinase 8
The duration of drug efficacy in
vivo is a key aspect primarily
addressed during the lead optimization phase of drug discovery. Hence,
the availability of robust computational approaches that can predict
the residence time of a compound at its target would accelerate candidate
selection. Nowadays the theoretical prediction of this parameter is
still very challenging. Starting from methods reported in the literature,
we set up and validated a new metadynamics (META-D)-based protocol
that was used to rank the experimental residence times of 10 arylpyrazole
cyclin-dependent kinase 8 (CDK8) inhibitors for which target-bound
X-ray structures are available. The application of reported methods
based on the detection of the escape from the first free energy well
gave a poor correlation with the experimental values. Our protocol
evaluates the energetics of the whole unbinding process, accounting
for multiple intermediates and transition states. Using seven collective
variables (CVs) encoding both roto-translational and conformational
motions of the ligand, a history-dependent biasing potential is deposited
as a sum of constant-height Gaussian functions until the ligand reaches
an unbound state. The time required to achieve this state is proportional
to the integral of the deposited potential over the CV hyperspace.
Average values of this time, for replicated META-D simulations, provided
an accurate classification of CDK8 inhibitors spanning short, medium,
and long residence times
Fatty Acid Amide Hydrolase (FAAH), Acetylcholinesterase (AChE), and Butyrylcholinesterase (BuChE): Networked Targets for the Development of Carbamates as Potential Anti-Alzheimer’s Disease Agents
The modulation of the endocannabinoid
system is emerging as a viable
avenue for the treatment of neurodegeneration, being involved in neuroprotective
and anti-inflammatory processes. In particular, indirectly enhancing
endocannabinoid signaling to therapeutic levels through FAAH inhibition
might be beneficial for neurodegenerative disorders such as Alzheimer’s
disease, effectively preventing or slowing the progression of the
disease. Hence, in the search for a more effective treatment for Alzheimer’s
disease, in this paper, the multitarget-directed ligand paradigm was
applied to the design of carbamates able to simultaneously target
the recently proposed endocannabinoid system and the classic cholinesterase
system, and achieve effective dual FAAH/cholinesterase inhibitors.
Among the two series of synthesized compounds, while some derivatives
proved to be extremely potent on a single target, compounds <b>9</b> and <b>19</b> were identified as effective dual FAAH/ChE
inhibitors, with well-balanced nanomolar activities. Thus, <b>9</b> and <b>19</b> might be considered as new promising candidates
for Alzheimer’s disease treatment
Fatty Acid Amide Hydrolase (FAAH), Acetylcholinesterase (AChE), and Butyrylcholinesterase (BuChE): Networked Targets for the Development of Carbamates as Potential Anti-Alzheimer’s Disease Agents
The modulation of the endocannabinoid
system is emerging as a viable
avenue for the treatment of neurodegeneration, being involved in neuroprotective
and anti-inflammatory processes. In particular, indirectly enhancing
endocannabinoid signaling to therapeutic levels through FAAH inhibition
might be beneficial for neurodegenerative disorders such as Alzheimer’s
disease, effectively preventing or slowing the progression of the
disease. Hence, in the search for a more effective treatment for Alzheimer’s
disease, in this paper, the multitarget-directed ligand paradigm was
applied to the design of carbamates able to simultaneously target
the recently proposed endocannabinoid system and the classic cholinesterase
system, and achieve effective dual FAAH/cholinesterase inhibitors.
Among the two series of synthesized compounds, while some derivatives
proved to be extremely potent on a single target, compounds <b>9</b> and <b>19</b> were identified as effective dual FAAH/ChE
inhibitors, with well-balanced nanomolar activities. Thus, <b>9</b> and <b>19</b> might be considered as new promising candidates
for Alzheimer’s disease treatment
Atropisomerism and Conformational Equilibria: Impact on PI3Kδ Inhibition of 2‑((6-Amino‑9<i>H</i>‑purin-9-yl)methyl)-5-methyl-3‑(<i>o</i>‑tolyl)quinazolin-4(3<i>H</i>)‑one (IC87114) and Its Conformationally Restricted Analogs
IC87114
[compound <b>1</b>, (2-((6-amino-9<i>H</i>-purin-9-yl)Âmethyl)-5-methyl-3-(<i>o</i>-tolyl)Âquinazolin-4Â(3<i>H</i>)-one)] is
a potent PI3K inhibitor selective for the δ
isoform. As predicted by molecular modeling calculations, rotation
around the bond connecting the quinazolin-4Â(3<i>H</i>)-one
nucleus to the <i>o</i>-tolyl is sterically hampered, which
leads to separable conformers with axial chirality (i.e., atropisomers).
After verifying that the a<i>S</i> and a<i>R</i> isomers of compound <b>1</b> do not interconvert in solution,
we investigated how biological activity is influenced by axial chirality
and conformational equilibrium. The a<i>S</i> and a<i>R</i> atropisomers of <b>1</b> were equally active in
the PI3Kδ assay. Conversely, the introduction of a methyl group
at the methylene hinge connecting the 6-amino-9<i>H</i>-purin-9-yl
pendant to the quinazolin-4Â(3<i>H</i>)-one nucleus of both
a<i>S</i> and a<i>R</i> isomers of <b>1</b> had a critical effect on the inhibitory activity, indicating that
modulation of the conformational space accessible for the two bonds
departing from the central methylene considerably affects the binding
of compound <b>1</b> analogues to PI3Kδ enzyme
Atropisomerism and Conformational Equilibria: Impact on PI3Kδ Inhibition of 2‑((6-Amino‑9<i>H</i>‑purin-9-yl)methyl)-5-methyl-3‑(<i>o</i>‑tolyl)quinazolin-4(3<i>H</i>)‑one (IC87114) and Its Conformationally Restricted Analogs
IC87114
[compound <b>1</b>, (2-((6-amino-9<i>H</i>-purin-9-yl)Âmethyl)-5-methyl-3-(<i>o</i>-tolyl)Âquinazolin-4Â(3<i>H</i>)-one)] is
a potent PI3K inhibitor selective for the δ
isoform. As predicted by molecular modeling calculations, rotation
around the bond connecting the quinazolin-4Â(3<i>H</i>)-one
nucleus to the <i>o</i>-tolyl is sterically hampered, which
leads to separable conformers with axial chirality (i.e., atropisomers).
After verifying that the a<i>S</i> and a<i>R</i> isomers of compound <b>1</b> do not interconvert in solution,
we investigated how biological activity is influenced by axial chirality
and conformational equilibrium. The a<i>S</i> and a<i>R</i> atropisomers of <b>1</b> were equally active in
the PI3Kδ assay. Conversely, the introduction of a methyl group
at the methylene hinge connecting the 6-amino-9<i>H</i>-purin-9-yl
pendant to the quinazolin-4Â(3<i>H</i>)-one nucleus of both
a<i>S</i> and a<i>R</i> isomers of <b>1</b> had a critical effect on the inhibitory activity, indicating that
modulation of the conformational space accessible for the two bonds
departing from the central methylene considerably affects the binding
of compound <b>1</b> analogues to PI3Kδ enzyme
Amino Acid Derivatives as Palmitoylethanolamide Prodrugs: Synthesis, <i>In Vitro</i> Metabolism and <i>In Vivo</i> Plasma Profile in Rats
<div><p>Palmitoylethanolamide (PEA) has antinflammatory and antinociceptive properties widely exploited in veterinary and human medicine, despite its poor pharmacokinetics. Looking for prodrugs that could progressively release PEA to maintain effective plasma concentrations, we prepared carbonates, esters and carbamates at the hydroxyl group of PEA. Chemical stability (pH 7.4) and stability in rat plasma and liver homogenate were evaluated by in vitro assays. Carbonates and carbamates resulted too labile and too resistant in plasma, respectively. Ester derivatives, prepared by conjugating PEA with various amino acids, allowed to modulate the kinetics of PEA release in plasma and stability in liver homogenate. L-Val-PEA, with suitable PEA release in plasma, and D-Val-PEA, with high resistance to hepatic degradation, were orally administered to rats and plasma levels of prodrugs and PEA were measured at different time points. Both prodrugs showed significant release of PEA, but provided lower plasma concentrations than those obtained with equimolar doses of PEA. Amino-acid esters of PEA are a promising class to develop prodrugs, even if they need further chemical optimization.</p></div