ABSTRACT
COT (Tpl2 in mice) is a serine/threonine MAP3 kinase that regulates production of TNF-alpha and other pro-inflammatory cytokines such as IL-1beta via the ERK/MAP kinase pathway. As TNF-alpha and IL-1beta are clinically validated targets for therapeutic intervention in rheumatoid arthritis (RA), blocking COT provides a potential avenue for amelioration of disease. Herein we describe identification of a cellular active selective small molecule inhibitor of COT kinase.
Subject(s)
Enzyme Inhibitors/chemical synthesis , Enzyme Inhibitors/pharmacology , MAP Kinase Kinase Kinases/antagonists & inhibitors , Proto-Oncogene Proteins/antagonists & inhibitors , Pyridines/chemical synthesis , Tumor Necrosis Factor-alpha/antagonists & inhibitors , Animals , Arthritis, Rheumatoid/drug therapy , Chemistry, Pharmaceutical/methods , Drug Design , Extracellular Signal-Regulated MAP Kinases/metabolism , Humans , Hydrogen Bonding , Inhibitory Concentration 50 , Interleukin-1beta/metabolism , Ligands , MAP Kinase Kinase Kinases/chemistry , Mice , Molecular Structure , Proto-Oncogene Proteins/chemistry , Pyridines/pharmacology , Tumor Necrosis Factor-alpha/chemistry , Tumor Necrosis Factor-alpha/metabolismABSTRACT
Evaluation of hit chemotypes from high throughput screening identified a novel series of 2,4-disubstituted thieno[2,3-c]pyridines as COT kinase inhibitors. Structural modifications exploring SAR at the 2- and 4-positions resulting in inhibitors with improved enzyme potency and cellular activity are disclosed.
Subject(s)
MAP Kinase Kinase Kinases/antagonists & inhibitors , Models, Molecular , Proto-Oncogene Proteins/antagonists & inhibitors , Pyridines/chemical synthesis , Pyridines/pharmacology , Thiophenes/chemical synthesis , Thiophenes/pharmacology , Combinatorial Chemistry Techniques , Crystallography, X-Ray , Humans , Molecular Conformation , Molecular Structure , Pyridines/chemistry , Structure-Activity Relationship , Thiophenes/chemistryABSTRACT
A series of benzoisoxazoles and benzoisothiazoles have been synthesized and evaluated as inhibitors of receptor tyrosine kinases (RTKs). Structure-activity relationship studies led to the identification of 3-amino benzo[ d]isoxazoles, incorporating a N, N'-diphenyl urea moiety at the 4-position that potently inhibited both the vascular endothelial growth factor receptor (VEGFR) and platelet-derived growth factor receptor families of RTKs. Within this series, orally bioavailable compounds possessing promising pharmacokinetic profiles were identified, and a number of compounds demonstrated in vivo efficacy in models of VEGF-stimulated vascular permeability and tumor growth. In particular, compound 50 exhibited an ED 50 of 2.0 mg/kg in the VEGF-stimulated uterine edema model and 81% inhibition in the human fibrosarcoma (HT1080) tumor growth model when given orally at a dose of 10 mg/kg/day.
Subject(s)
Isoxazoles/chemical synthesis , Models, Molecular , Oxazoles/chemical synthesis , Phenylurea Compounds/chemical synthesis , Receptors, Platelet-Derived Growth Factor/antagonists & inhibitors , Receptors, Vascular Endothelial Growth Factor/antagonists & inhibitors , Animals , Anti-Inflammatory Agents, Non-Steroidal/chemical synthesis , Anti-Inflammatory Agents, Non-Steroidal/pharmacokinetics , Anti-Inflammatory Agents, Non-Steroidal/pharmacology , Antineoplastic Agents/chemical synthesis , Antineoplastic Agents/pharmacokinetics , Antineoplastic Agents/pharmacology , Binding Sites , Biological Availability , Capillary Permeability/drug effects , Cell Line , Cell Line, Tumor , Edema/drug therapy , Female , Humans , Isoxazoles/pharmacokinetics , Isoxazoles/pharmacology , Mice , Mice, Inbred BALB C , NIH 3T3 Cells , Oxazoles/pharmacokinetics , Oxazoles/pharmacology , Phenylurea Compounds/pharmacokinetics , Phenylurea Compounds/pharmacology , Phosphorylation , Structure-Activity Relationship , Uterus/blood supply , Xenograft Model Antitumor AssaysABSTRACT
In our continued efforts to search for potent and novel receptor tyrosine kinase (RTK) inhibitors as potential anticancer agents, we discovered, through a structure-based design, that 3-aminoindazole could serve as an efficient hinge-binding template for kinase inhibitors. By incorporating an N,N'-diaryl urea moiety at the C4-position of 3-aminodazole, a series of RTK inhibitors were generated, which potently inhibited the tyrosine kinase activity of the vascular endothelial growth factor receptor and the platelet-derived growth factor receptor families. A number of compounds with potent oral activity were identified by utilizing an estradiol-induced mouse uterine edema model and an HT1080 human fibrosarcoma xenograft tumor model. In particular, compound 17p (ABT-869) was found to possess favorable pharmacokinetic profiles across different species and display significant tumor growth inhibition in multiple preclinical animal models.
Subject(s)
Angiogenesis Inhibitors/chemical synthesis , Indazoles/chemical synthesis , Phenylurea Compounds/chemical synthesis , Receptor Protein-Tyrosine Kinases/antagonists & inhibitors , Adenosine Triphosphate/chemistry , Administration, Oral , Angiogenesis Inhibitors/chemistry , Angiogenesis Inhibitors/pharmacology , Animals , Binding Sites , Edema/chemically induced , Edema/pathology , Estradiol , Female , Humans , Hydrophobic and Hydrophilic Interactions , Indazoles/chemistry , Indazoles/pharmacology , Male , Mice , Models, Molecular , NIH 3T3 Cells , Phenylurea Compounds/chemistry , Phenylurea Compounds/pharmacology , Phosphorylation , Receptor Protein-Tyrosine Kinases/chemistry , Receptor Protein-Tyrosine Kinases/metabolism , Structure-Activity Relationship , Uterus/drug effects , Uterus/pathology , Xenograft Model Antitumor AssaysABSTRACT
We report the synthesis and biological evaluation of 5-substituted 1,4-dihydroindeno[1,2-c]pyrazoles as multitargeted kinase inhibitors. Initial efforts focused on the development of selective KDR inhibitors, while later strategies involved the improvement of potency toward multiple kinase targets. Thus, several compounds were identified as potent KDR, Flt1, Flt3, and c-Kit inhibitors.
Subject(s)
Protein Kinase Inhibitors/chemistry , Protein Kinase Inhibitors/pharmacology , Pyrazoles/chemistry , Pyrazoles/pharmacology , Receptor Protein-Tyrosine Kinases/antagonists & inhibitors , Animals , Humans , Molecular Structure , Protein Kinase Inhibitors/chemical synthesis , Pyrazoles/chemical synthesisABSTRACT
A series of substituted thienopyridine ureas was prepared and evaluated for enzymatic and cellular inhibition of KDR kinase activity. Several of these analogs, such as 2, are potent inhibitors of KDR (<10 nM) in both enzymatic and cellular assays. Further characterization of inhibitor 2 indicated that this analog possessed excellent in vivo potency (ED50 2.1 mg/kg) as measured in an estradiol-induced mouse uterine edema model.
Subject(s)
Pyridines/chemical synthesis , Urea/chemical synthesis , Vascular Endothelial Growth Factor Receptor-2/antagonists & inhibitors , Animals , Disease Models, Animal , Edema/chemically induced , Estradiol , Female , Mice , Models, Molecular , Pyridines/pharmacology , Structure-Activity Relationship , Urea/chemistry , Urea/pharmacology , Uterine Diseases/pathologyABSTRACT
A series of isothiazolopyrimidines and isoxazolopyrimidines were synthesized and identified as potent KDR inhibitors. SAR studies led to isothiazolopyrimidine urea analogs that potently inhibit VEGFR tyrosine kinases (KDR enzymatic and cellular IC(50) values below 10 nM) as well as cKIT and TIE2. The selected compounds 8 and 13 display 56% and 48% oral bioavailability in mice, respectively.
Subject(s)
Enzyme Inhibitors/pharmacology , Pyrimidines/chemistry , Receptor Protein-Tyrosine Kinases/antagonists & inhibitors , Administration, Oral , Animals , Enzyme Inhibitors/chemistry , Humans , Inhibitory Concentration 50 , Kinetics , Mice , Models, Chemical , Models, Molecular , Proto-Oncogene Proteins c-kit/metabolism , Pyrimidines/pharmacology , Receptor Protein-Tyrosine Kinases/chemistry , Receptor, TIE-2/metabolism , Structure-Activity Relationship , Vascular Endothelial Growth Factor Receptor-1/metabolismABSTRACT
A series of novel thienopyrimidine-based receptor tyrosine kinase inhibitors has been discovered. Investigation of structure-activity relationships at the 5- and 6-positions of the thienopyrimidine nucleus led to a series of N,N'-diaryl ureas that potently inhibit all of the vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF) receptor tyrosine kinases. A kinase insert domain-containing receptor (KDR) homology model suggests that these compounds bind to the "inactive conformation" of the enzyme with the urea portion extending into the back hydrophobic pocket adjacent to the adenosine 5'-triphosphate (ATP) binding site. A number of compounds have been identified as displaying excellent in vivo potency. In particular, compounds 28 and 76 possess favorable pharmacokinetic (PK) profiles and demonstrate potent antitumor efficacy against the HT1080 human fibrosarcoma xenograft tumor growth model (tumor growth inhibition (TGI) = 75% at 25 mg/kg.day, per os (po)).
Subject(s)
Antineoplastic Agents/chemical synthesis , Pyrimidines/chemical synthesis , Receptors, Platelet-Derived Growth Factor/antagonists & inhibitors , Urea/analogs & derivatives , Urea/chemical synthesis , Vascular Endothelial Growth Factor Receptor-2/antagonists & inhibitors , Adenosine Triphosphate/chemistry , Animals , Antineoplastic Agents/chemistry , Antineoplastic Agents/pharmacology , Edema/chemically induced , Edema/pathology , Estradiol , Female , Humans , Mice , Mice, Inbred BALB C , Mice, SCID , Models, Molecular , NIH 3T3 Cells , Phosphorylation , Pyrimidines/chemistry , Pyrimidines/pharmacology , Structure-Activity Relationship , Urea/chemistry , Urea/pharmacology , Uterus/drug effects , Uterus/pathology , Vascular Endothelial Growth Factor Receptor-2/chemistry , Vascular Endothelial Growth Factor Receptor-2/metabolism , Xenograft Model Antitumor AssaysABSTRACT
A series of substituted isoindolinone ureas was prepared and evaluated for enzymatic and cellular inhibition of KDR kinase activity. Several of these analogs, such as 14c, are potent inhibitors of KDR both enzymatically (< 50 nM) and cellularly < or = 100 nM). A 3D KDR/CDK2/MAP kinase overlay model with several structurally related tyrosine kinase inhibitors was used to predict the binding interactions of the isoindolinone ureas with the KDR active site.