ABSTRACT
Compounds with serotonin reuptake inhibition/5-HT(1A) dual activity were used to build 3D pharmacophore model as a training molecules by Discover Studio. Based on the model, 8 novel aryl piperazine benzo[b][1,4] oxazine derivatives were designed and synthesized, and their structures were confirmed by 1H NMR and HR-MS. Biological evaluation illustrated that compounds VI(1) and VI(7) showed potent functional activities at both 5-HT transporter and 5-HT(1A) receptor, which can be used as lead compounds to guide future research of design and synthesis of potent novel compounds.
Subject(s)
Animals , Cricetinae , CHO Cells , Cricetulus , Drug Design , Genetic Vectors , Molecular Structure , Oxazines , Chemistry , Pharmacology , Piperazines , Chemistry , Pharmacology , Plasmids , Protein Binding , Receptor, Serotonin, 5-HT1A , Genetics , Metabolism , Serotonin Plasma Membrane Transport Proteins , Genetics , Metabolism , Selective Serotonin Reuptake Inhibitors , Metabolism , Structure-Activity Relationship , TransfectionABSTRACT
<p><b>OBJECTIVE</b>To explore the possible differential trafficking properties of the dopamine D1-like receptor subtypes, D1 receptor and D5 receptor.</p><p><b>METHODS</b>To visualize distributions of dopamine D1-like receptor subtypes at subcellular level, the yellow and cyan variants of green fluorescent protein (GFP) were used to tag D1 and D5 receptors. After transfection with the tagged dopamine receptors, the neuroblastoma cells NG108-15 were treated with D1 agonist SKF38393 or acetylcholine (ACh). Then we observed the subcellular distributions of the tagged receptors under the confocal microscopy and tried to determine trafficking properties by comparing their distribution patterns before and after the drug treatment.</p><p><b>RESULTS</b>In resting conditions, D1 receptors located in the plasma membrane of NG108-15 cells, while D5 receptors located in both plasma membrane and cytosol. With the pre-treatment of SKF38393, the subcellular distribution of D1 receptors was changed. The yellow particle-like fluorescence of tagged D1 receptors appeared in the cytosol, indicating that D1 receptors were internalized into cytosol from the cell surface. Same situation also occurred in ACh pre-treatment. In contrast, the subcellular distribution of D5 receptors was not changed after SKF38393 or ACh treatment, indicating that D5R was not translocated to cell surface. Interestingly, when D1 and D5 receptors were co-expressed in the same cell, both kept their distinct subcellular distribution patterns and the trafficking properties.</p><p><b>CONCLUSION</b>Our present study reveals that in NG108-15 nerve cells, dopamine D1 and D5 receptors exhibit differential subcellular distribution patterns, and only D1 receptor has a marked trafficking response to the drug stimulation. We further discuss the potential role of the differential trafficking properties of D1-like receptors in complex modulation of DA signaling.</p>
Subject(s)
Animals , Humans , Mice , Rats , 2,3,4,5-Tetrahydro-7,8-dihydroxy-1-phenyl-1H-3-benzazepine , Pharmacology , Acetylcholine , Pharmacology , Cell Line , Dopamine Agonists , Pharmacology , HeLa Cells , Luminescent Proteins , Genetics , Microscopy, Confocal , Methods , Neuroblastoma , Protein Transport , Receptors, Dopamine D1 , Metabolism , Receptors, Dopamine D5 , Metabolism , Subcellular Fractions , Metabolism , Transfection , MethodsABSTRACT
A series of nitrogen-containing benzoheterocyclic derivatives were synthesized and tested for their antipsychotic activities. Their structures were confirmed by 1H NMR and HR-MS. Preliminary in vitro pharmacological trials showed that most of the target compounds have high affinity with D2 and 5-HT(2A) receptors. Among the tested compounds, 20 exhibited the highest affinity and D2 partial agonist activity. In vivo studies showed 20 has potent antipsychotic activities on apomorphine mice model, which is a chance to find a better precursor of D2 partial agonist for further optimization.