ABSTRACT
High-throughput screening for the induction of a luciferase reporter gene in a thrombopoietin (TPO)-responsive cell line resulted in the identification of 4-diazo-3-hydroxy-1-naphthalenesulfonic acids as TPO mimics. Modification of the core structure and adjustment of unwanted functionality resulted in the development of (5-oxo-1,5-dihydropyrazol-4-ylidene)hydrazines which exhibited efficacies equivalent to those of TPO in several cell-based assays designed to measure thrombopoietic activity. Furthermore, these compounds elicited biochemical responses in TPO-receptor-expressing cells similar to those in TPO itself, including kinase activation and protein phosphorylation. Potencies for the best compounds were high for such low molecular weight compounds (MW < 500) with EC(50) values in the region of 1-20 nM.
Subject(s)
Azo Compounds/chemical synthesis , Hydrazines/chemical synthesis , Megakaryocytes/drug effects , Naphthalenesulfonates/chemical synthesis , Neoplasm Proteins , Pyrazoles/chemical synthesis , Receptors, Cytokine , Thrombopoietin/chemistry , Animals , Azo Compounds/chemistry , Azo Compounds/pharmacology , Cell Division , Cell Line , Drug Evaluation, Preclinical , Electrophoretic Mobility Shift Assay , Enzyme Activation , Genes, Reporter , Hydrazines/chemistry , Hydrazines/pharmacology , Luciferases/genetics , Luciferases/metabolism , Mice , Molecular Mimicry , Molecular Weight , Naphthalenesulfonates/chemistry , Naphthalenesulfonates/pharmacology , Phosphorylation , Phosphotransferases/metabolism , Proto-Oncogene Proteins/metabolism , Pyrazoles/chemistry , Pyrazoles/pharmacology , Receptors, Thrombopoietin , Structure-Activity Relationship , Thrombopoietin/metabolismABSTRACT
The discovery of a series of phenylalanine derived CCR3 antagonists is reported. Parallel, solution-phase library synthesis has been utilized to delineate the structure-activity relationship leading to the synthesis of highly potent, CCR3-selective antagonists.
Subject(s)
Phenylalanine/chemistry , Phenylalanine/pharmacology , Receptors, Chemokine/antagonists & inhibitors , Humans , Receptors, CCR3 , Receptors, Chemokine/metabolism , Structure-Activity RelationshipABSTRACT
Highly potent CCR3 antagonists have been developed from a previously reported series of phenylalanine ester-based leads. Solution-phase, parallel synthesis optimization was utilized to identify highly potent, functional CCR3 antagonists.
Subject(s)
Phenylalanine/pharmacology , Receptors, Chemokine/antagonists & inhibitors , Humans , Phenylalanine/chemical synthesis , Phenylalanine/chemistry , Receptors, CCR3 , Receptors, Chemokine/metabolism , Structure-Activity RelationshipABSTRACT
A versatile synthetic route to a novel series of bis-imidazolemethanes designed to inhibit the hCMV protease has been developed and a series of potential metal binding inhibitors has been identified. In selectivity assays, the compounds were highly specific for CMV protease and showed no inhibition (IC50 > 100 microM) of other prototypical serine proteases such as trypsin, elastase, and chymotrypsin. Although the presence of free zinc ions was found to be an absolute requirement for the in vitro biological activity of this class of inhibitor, the potency of the inhibitors could not be improved beyond the micromolar level.
Subject(s)
Imidazoles/chemical synthesis , Serine Endopeptidases/metabolism , Serine Proteinase Inhibitors/chemical synthesis , Chymotrypsin/antagonists & inhibitors , Drug Design , Humans , Imidazoles/chemistry , Metals , Molecular Conformation , Molecular Structure , Pancreatic Elastase/antagonists & inhibitors , Serine Proteinase Inhibitors/chemistry , Serine Proteinase Inhibitors/pharmacology , Structure-Activity Relationship , Trypsin/metabolismABSTRACT
Eosinophils have been implicated in the pathogenesis of asthma and other allergic diseases. Several CC chemokines including eotaxin (CCL-11), eotaxin-2 (CCL-24), RANTES (CCL-5), and monocyte chemotactic protein-3 (MCP-3, CCL-7) and 4 (MCP-4, CCL-13) are potent eosinophil chemotactic and activating peptides acting through CC chemokine receptor-3 (CCR3). Thus, antagonism of CCR3 could have a therapeutic role in asthma and other eosinophil-mediated diseases. A high throughput, cellular functional screen was configured using RBL-2H3 cells stably expressing CCR3 (RBL-2H3-CCR3) to identify non-peptide receptor antagonists. A small molecule CCR3 antagonist was identified, SK&F 45523, and chemical optimization led to the generation of a number of highly potent, selective CCR3 antagonists including SB-297006 and SB-328437. These compounds were further characterized in vitro and demonstrated high affinity, competitive inhibition of (125)I-eotaxin and (125)I-MCP-4 binding to human eosinophils. The compounds were potent inhibitors of eotaxin- and MCP-4-induced Ca(2+) mobilization in RBL-2H3-CCR3 cells and eosinophils. Additionally, SB-328437 inhibited eosinophil chemotaxis induced by three ligands that activate CCR3 with similar potencies. Selectivity was affirmed using a panel of 10 seven-transmembrane receptors. This is the first description of a non-peptide CCR3 antagonist, which should be useful in further elucidating the pathophysiological role of CCR3 in allergic inflammatory diseases.
Subject(s)
Benzamides/pharmacology , Cell Movement/drug effects , Chemokines, CC/antagonists & inhibitors , Cytokines/antagonists & inhibitors , Eosinophils/drug effects , Monocyte Chemoattractant Proteins/antagonists & inhibitors , Naphthalenes/pharmacology , Phenylalanine/analogs & derivatives , Receptors, Chemokine/antagonists & inhibitors , Receptors, HIV/antagonists & inhibitors , Asthma/physiopathology , Binding, Competitive , Calcium/metabolism , Cell Line , Chemokine CCL11 , Chemokine CCL24 , Humans , Phenylalanine/pharmacology , Receptors, CCR3 , Receptors, Chemokine/physiologyABSTRACT
A novel class of CMV protease inhibitors based on a benzothiopyran-S,S-dioxide nucleus has been discovered. Enzyme kinetic data supports a reversible mode of inhibition for a representative member of this class, 2-(3-pyridyl-N-oxide)benzothiopyran-4-one-S,S-dioxide, 1. Experiments in the presence and absence of the disulfide reducing agent DTT suggest that the inhibition by 1 is not due to oxidative inactivation of the enzyme. Also presented are results of some SAR studies of the benzothiopyranone ring system.
