ABSTRACT
Using structure-based design, a new class of inhibitors of protein tyrosine phosphatase-1B (PTP1B) has been identified, which incorporate the 1,2,5-thiadiazolidin-3-one-1,1-dioxide template.
Subject(s)
Enzyme Inhibitors/pharmacology , Protein Tyrosine Phosphatases/antagonists & inhibitors , Thiadiazines/chemistry , Thiadiazines/pharmacology , Crystallography, X-Ray , Drug Design , Enzyme Inhibitors/chemistry , Ligands , Magnetic Resonance Spectroscopy , Molecular Conformation , Protein Tyrosine Phosphatase, Non-Receptor Type 1 , Structure-Activity RelationshipABSTRACT
Modification of imidazo[1,2-a]pyridine CDK inhibitors lead to identification of less lipophilic imidazo[1,2-b]pyridazine series of CDK inhibitors. Although several equivalent compounds from these two series have similar structure and show similar CDK activity, the SAR of the two series differs significantly. Protein inhibitor structure determination has confirmed differences in binding mode and given some understanding of these differences in SAR. Potent and selective imidazo[1,2-b]pyridazine inhibitors of CDK2 have been identified, which show >1 microM plasma levels following a 2mg/kg oral dose to mice.
Subject(s)
Cyclin-Dependent Kinases/antagonists & inhibitors , Enzyme Inhibitors/pharmacology , Pyridazines/chemistry , Pyridazines/pharmacology , Animals , Enzyme Inhibitors/blood , Enzyme Inhibitors/chemistry , Mice , Models, Molecular , Pyridazines/bloodABSTRACT
Human thymidine phosphorylase (HTP), also known as platelet-derived endothelial cell growth factor (PD-ECGF), is overexpressed in certain solid tumors where it is linked to poor prognosis. HTP expression is utilized for certain chemotherapeutic strategies and is also thought to play a role in tumor angiogenesis. We determined the structure of HTP bound to the small molecule inhibitor 5-chloro-6-[1-(2-iminopyrrolidinyl) methyl] uracil hydrochloride (TPI). The inhibitor appears to mimic the substrate transition state, which may help explain the potency of this inhibitor and the catalytic mechanism of pyrimidine nucleotide phosphorylases (PYNPs). Further, we have confirmed the validity of the HTP structure as a template for structure-based drug design by predicting binding affinities for TPI and other known HTP inhibitors using in silico docking techniques. This work provides the first structural insight into the binding mode of any inhibitor to this important drug target and forms the basis for designing novel inhibitors for use in anticancer therapy.
Subject(s)
Models, Molecular , Protein Binding , Protein Folding , Pyrrolidines/chemistry , Thymidine Phosphorylase/metabolism , Uracil/analogs & derivatives , Uracil/chemistry , Crystallization , Enzyme Inhibitors/chemistry , Enzyme Inhibitors/pharmacology , Humans , Protein Structure, Tertiary , Pyrrolidines/pharmacology , Uracil/pharmacologyABSTRACT
Using a high-throughput screening campaign, we identified the 4,6-bis anilino pyrimidines as inhibitors of the cyclin-dependent kinase, CDK4. Herein we describe the further chemical modification and use of X-ray crystallography to develop potent and selective in vitro inhibitors of CDK4.
Subject(s)
Antineoplastic Agents/pharmacology , Cyclin-Dependent Kinases/antagonists & inhibitors , Proto-Oncogene Proteins , Pyrimidines/pharmacology , Animals , Antineoplastic Agents/chemical synthesis , Binding Sites , Crystallography, X-Ray , Cyclin-Dependent Kinase 4 , Humans , Inhibitory Concentration 50 , Molecular Structure , Pyrimidines/chemical synthesis , Structure-Activity RelationshipABSTRACT
Through chemical modification and X-ray crystallography we identified the 2,4-bis anilino pyrimidines as potent inhibitors of CDK4. Herein, we describe the optimisation of this series.
Subject(s)
Antineoplastic Agents/pharmacology , Cyclin-Dependent Kinases/antagonists & inhibitors , Proto-Oncogene Proteins , Pyrimidines/pharmacology , Adenosine Triphosphate/chemistry , Animals , Antineoplastic Agents/chemical synthesis , Binding Sites , Crystallography, X-Ray , Cyclin-Dependent Kinase 4 , Humans , Inhibitory Concentration 50 , Molecular Structure , Pyrimidines/chemical synthesis , Structure-Activity RelationshipABSTRACT
High-throughput screening identified the imidazo[1,2-a]pyridine and bisanilinopyrimidine series as inhibitors of the cyclin-dependent kinase CDK4. Comparison of their experimentally-determined binding modes and emerging structure-activity trends led to the development of potent and selective imidazo[1,2-a]pyridine inhibitors for CDK4 and in particular CDK2.
Subject(s)
Cyclin-Dependent Kinases/antagonists & inhibitors , Proto-Oncogene Proteins , Pyridines/chemical synthesis , CDC2-CDC28 Kinases/antagonists & inhibitors , CDC2-CDC28 Kinases/chemistry , Crystallography, X-Ray , Cyclin-Dependent Kinase 2 , Cyclin-Dependent Kinase 4 , Cyclin-Dependent Kinases/chemistry , Enzyme Inhibitors/chemical synthesis , Enzyme Inhibitors/chemistry , Enzyme Inhibitors/pharmacology , Humans , Inhibitory Concentration 50 , Molecular Structure , Protein Binding , Pyridines/chemistry , Pyridines/pharmacology , Structure-Activity RelationshipABSTRACT
Matrix metalloproteinases (MMPs) and their inhibitors are important in connective tissue re-modelling in diseases of the cardiovascular system, such as atherosclerosis. Various members of the MMP family have been shown to be expressed in atherosclerotic lesions, but MMP9 is consistently seen in inflammatory atherosclerotic lesions. MMP9 over-expression is implicated in the vascular re-modelling events preceding plaque rupture (the most common cause of acute myocardial infarction). Reduced MMP9 activity, either by genetic manipulation or through pharmacological intervention, has an impact on ventricular re-modelling following infarction. MMP9 activity may therefore represent a key mechanism in the pathogenesis of heart failure. We have determined the crystal structure, at 2.3 A resolution, of the catalytic domain of human MMP9 bound to a peptidic reverse hydroxamate inhibitor as well as the complex of the same inhibitor bound to an active-site mutant (E402Q) at 2.1 A resolution. MMP9 adopts the typical MMP fold. The catalytic centre is composed of the active-site zinc ion, co-ordinated by three histidine residues (401, 405 and 411) and the essential glutamic acid residue (402). The main differences between the catalytic domains of various MMPs occur in the S1' subsite or selectivity pocket. The S1' specificity site in MMP9 is perhaps best described as a tunnel leading toward solvent, as in MMP2 and MMP13, as opposed to the smaller pocket found in fibroblast collagenase and matrilysin. The present structure enables us to aid the design of potent and specific inhibitors for this important cardiovascular disease target.