ABSTRACT
A novel series of 1,2,4-triazolyl octahydropyrrolo[2,3-b]pyrroles showing high affinity and selectivity at the DA D3 receptor is reported here. Compounds endowed with high selectivity over the hERG channel were identified and their pharmacokinetic properties thoroughly analyzed. A few derivatives with appropriate developability characteristics were selected for further studies and progression along the screening cascade. In particular, derivative 60a, (DA D3 pKi=8.4, DA D2 pKi=6.0 and hERG fpKi=5.2) showed a balanced profile and further refinements are in progress around this molecule.
Subject(s)
Pyrroles/pharmacology , Receptors, Dopamine D3/antagonists & inhibitors , Triazoles/pharmacology , Animals , Binding, Competitive/drug effects , CHO Cells , Cricetulus , Dose-Response Relationship, Drug , Ether-A-Go-Go Potassium Channels/antagonists & inhibitors , HEK293 Cells , Humans , Models, Molecular , Molecular Structure , Pyrroles/chemical synthesis , Pyrroles/chemistry , Structure-Activity Relationship , Triazoles/chemical synthesis , Triazoles/chemistryABSTRACT
Further exploration around the recently disclosed potent triple re-uptake inhibitor 6-(3,4-dichlorophenyl)-1-[(methyloxy)methyl]-3-azabicyclo[4.1.0]heptane led to the identification of a new series of potent triple re-uptake inhibitors endowed with good developability characteristics. The insertion of a further aryl moiety into the template allowed the 'titration' of the SERT/NET/DAT ratio leading to the identification of further tools in this important area.
Subject(s)
Adrenergic Uptake Inhibitors/chemistry , Dopamine Uptake Inhibitors/chemistry , Heptanes/chemistry , Selective Serotonin Reuptake Inhibitors/chemistry , Adrenergic Uptake Inhibitors/chemical synthesis , Adrenergic Uptake Inhibitors/pharmacology , Aza Compounds/chemistry , Bridged Bicyclo Compounds/chemistry , Dopamine Plasma Membrane Transport Proteins/chemistry , Dopamine Plasma Membrane Transport Proteins/metabolism , Dopamine Uptake Inhibitors/chemical synthesis , Dopamine Uptake Inhibitors/pharmacology , Heptanes/chemical synthesis , Heptanes/pharmacology , Humans , Norepinephrine Plasma Membrane Transport Proteins/chemistry , Norepinephrine Plasma Membrane Transport Proteins/metabolism , Protein Binding , Serotonin Plasma Membrane Transport Proteins/chemistry , Serotonin Plasma Membrane Transport Proteins/metabolism , Selective Serotonin Reuptake Inhibitors/chemical synthesis , Selective Serotonin Reuptake Inhibitors/pharmacology , Structure-Activity RelationshipABSTRACT
IC87114 [compound 1, (2-((6-amino-9H-purin-9-yl)methyl)-5-methyl-3-(o-tolyl)quinazolin-4(3H)-one)] is a potent PI3K inhibitor selective for the δ isoform. As predicted by molecular modeling calculations, rotation around the bond connecting the quinazolin-4(3H)-one nucleus to the o-tolyl is sterically hampered, which leads to separable conformers with axial chirality (i.e., atropisomers). After verifying that the aS and aR isomers of compound 1 do not interconvert in solution, we investigated how biological activity is influenced by axial chirality and conformational equilibrium. The aS and aR atropisomers of 1 were equally active in the PI3Kδ assay. Conversely, the introduction of a methyl group at the methylene hinge connecting the 6-amino-9H-purin-9-yl pendant to the quinazolin-4(3H)-one nucleus of both aS and aR isomers of 1 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 1 analogues to PI3Kδ enzyme.
Subject(s)
Adenine/analogs & derivatives , Phosphoinositide-3 Kinase Inhibitors , Protein Kinase Inhibitors/chemistry , Protein Kinase Inhibitors/pharmacology , Quinazolines/chemistry , Quinazolines/pharmacology , Adenine/chemistry , Adenine/pharmacology , Animals , Class Ia Phosphatidylinositol 3-Kinase/chemistry , Class Ia Phosphatidylinositol 3-Kinase/metabolism , Humans , Isomerism , Mice , Models, MolecularABSTRACT
A novel series of 1,2,4-triazolyl 5-azaspiro[2.4]heptanes with high affinity and selectivity at the dopamine (DA) D3 receptor (D3R) is described. Some of these compounds also have high selectivity over the hERG channel and were characterized with respect to their pharmacokinetic properties both in vitro and in vivo during lead identification and early lead optimization phases. A few derivatives with overall favorable developability characteristics were selected for further late lead optimization studies.
Subject(s)
Heptanes/chemistry , Heptanes/pharmacology , Receptors, Dopamine D3/antagonists & inhibitors , Spiro Compounds/chemistry , Spiro Compounds/pharmacology , Animals , CHO Cells , Cricetulus , Crystallography, X-Ray , Ether-A-Go-Go Potassium Channels/metabolism , Humans , Models, Molecular , Receptors, Dopamine D3/metabolism , Structure-Activity Relationship , Triazoles/chemistry , Triazoles/pharmacologyABSTRACT
The characterisation of a microbial community of a polycyclic aromatic hydrocarbons (PAHs) contaminated site (formerly Carbochimica, Trento, Italy) was carried out. A preliminary evaluation of the heterogeneity and the metabolic activity of the microbial community were attempted by denaturing gradient gel electrophoresis (DGGE) and reverse transcriptase-denaturing gel electrophoresis (RT-DGGE). The presence of a heterogeneous and metabolically active microbial community was found. To evaluate the PAH-transforming potential of the soil bacterial community, enrichment cultures were set up. Taxonomically diverse bacteria, showing different biochemical PAH-transforming pathways were obtained. Some of the isolates showed not nah-homologous PAH-transforming genotypes.
Subject(s)
Bacteria/metabolism , Polycyclic Aromatic Hydrocarbons/metabolism , Soil Microbiology , Soil Pollutants/metabolism , Bacteria/genetics , Bacteria/growth & development , Bacteria/isolation & purification , Biodegradation, Environmental , Culture Media , DNA, Bacterial/genetics , Electrophoresis, Polyacrylamide Gel , Genes, Bacterial , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
There is preclinical evidence supporting the finding that the GABA(B) receptor orthosteric agonist, baclofen, has significant effects on eating behavior suggesting the potential therapeutic application of this compound for the treatment of eating related disorders. However, the wide clinical use of baclofen might be limited by the appearance of sedative and motor impairment effects. The identification of positive allosteric modulators (PAMs) of GABA(B) receptors represents a novel therapeutic approach to reduce the centrally-mediated adverse effects typical of the GABA(B) receptor orthosteric agonist. In the present work, we report the in vitro profile of a novel chemical structure, 2-{1-[2-(4-chlorophenyl)-5-methylpyrazolo[1,5-a]pyrimidin-7-yl]-2-piperidinyl}ethanol (CMPPE) identified by screening the GSK compound collection. CMPPE potentiates GABA-stimulated [(35)S]GTPγS binding to membranes of human recombinant cell line and of rat brain cortex. GABA concentration-response curves (CRC) in the presence of fixed concentrations of CMPPE, in rat native tissue, revealed an increase of both the potency and maximal efficacy of GABA. A similar modulatory effect was observed in GABA(B) receptor-mediated activation of inwardly rectifying potassium channels in hippocampal neurons. CMPPE (30-100 mg/kg) and GS39783 (100 mg/kg) significantly decreased food consumption in rat without impairment on the animal locomotor activity. On the contrary, baclofen (2.5 mg/kg) decreased both food intake and motor performance. All together these findings confirm the role of GABA(B) system in controlling animal food intake and for the first time demonstrate that GABA(B) receptor PAMs may represent a novel pharmacological approach to treat eating disorders without unwanted sedative effects.
Subject(s)
Baclofen/pharmacology , Cyclopentanes/pharmacology , GABA-B Receptor Agonists/pharmacology , Molecular Targeted Therapy , Pyrazoles/pharmacology , Pyrimidines/pharmacology , Receptors, GABA-B/metabolism , Animals , Brain/metabolism , CHO Cells , Cerebral Cortex/drug effects , Cerebral Cortex/metabolism , Cricetinae , Drug Evaluation, Preclinical , Eating/drug effects , Eating/physiology , Electrophysiological Phenomena , Hippocampus/drug effects , Hippocampus/metabolism , Humans , Male , Membrane Proteins/analysis , Motor Activity/drug effects , Neurons/drug effects , Neurons/metabolism , Pyrazoles/chemistry , Pyrimidines/chemistry , Radioligand Assay , Rats , Rats, Sprague-Dawley , Receptors, GABA-B/drug effects , Receptors, GABA-B/genetics , Transfection , gamma-Aminobutyric Acid/physiologyABSTRACT
The discovery of new highly potent and selective triple reuptake inhibitors is reported. The new classes of 1-(aryl)-6-[alkoxyalkyl]-3-azabicyclo[3.1.0]hexanes and 6-(aryl)-6-[alkoxyalkyl]-3-azabicyclo[3.1.0]hexanes are described together with detailed SAR. Appropriate decoration of the scaffolds was achieved with the help of a triple reuptake inhibitor pharmacophore model detailed here. Selected derivatives showed good oral bioavailability (>30%) and brain penetration (B/B > 4) in rats associated with high in vitro potency and selectivity at SERT, NET, and DAT. Among these compounds, microdialysis and in vivo experiments confirm that derivative 15 has an appropriate developability profile to be considered for further progression.
Subject(s)
Azabicyclo Compounds/pharmacology , Dopamine Plasma Membrane Transport Proteins/antagonists & inhibitors , Norepinephrine Plasma Membrane Transport Proteins/antagonists & inhibitors , Serotonin Plasma Membrane Transport Proteins/metabolism , Animals , Azabicyclo Compounds/chemistry , Azabicyclo Compounds/pharmacokinetics , Binding, Competitive , Biogenic Monoamines/metabolism , Biological Availability , Biological Transport/drug effects , Cell Line , Chromatography, High Pressure Liquid , Cytochrome P-450 Enzyme Inhibitors , Cytochrome P-450 Enzyme System/metabolism , Dopamine Plasma Membrane Transport Proteins/metabolism , Humans , Male , Mice , Microdialysis , Microsomes, Liver/metabolism , Models, Chemical , Molecular Structure , Motor Activity/drug effects , Norepinephrine Plasma Membrane Transport Proteins/metabolism , Prefrontal Cortex/metabolism , Rats , Structure-Activity RelationshipABSTRACT
A pharmacophore model for triple reuptake inhibitors and the new class of 1-(aryl)-6-[alkoxyalkyl]-3-azabicyclo[3.1.0]hexanes were recently reported. Further investigation in this area led to the identification of a new series of potent and selective triple reuptake inhibitors endowed with good developability characteristics. Excellent bioavailability and brain penetration are associated with this series of 6-(3,4-dichlorophenyl)-1-[(methyloxy)methyl]-3-azabicyclo[4.1.0]heptanes together with high in vitro potency and selectivity at SERT, NET, and DAT. In vivo microdialysis experiments in different animal models and receptor occupancy studies in rat confirmed that derivative 17 showed an appropriate profile to guarantee further progression of the compound.