ABSTRACT
The nuclear export protein XPO1 is overexpressed in cancer, leading to the cytoplasmic mislocalization of multiple tumor suppressor proteins. Existing XPO1-targeting agents lack selectivity and have been associated with significant toxicity. Small molecule selective inhibitors of nuclear export (SINEs) were designed that specifically inhibit XPO1. Genetic experiments and X-ray structures demonstrate that SINE covalently bind to a cysteine residue in the cargo-binding groove of XPO1, thereby inhibiting nuclear export of cargo proteins. The clinical relevance of SINEs was explored in chronic lymphocytic leukemia (CLL), a disease associated with recurrent XPO1 mutations. Evidence is presented that SINEs can restore normal regulation to the majority of the dysregulated pathways in CLL both in vitro and in vivo and induce apoptosis of CLL cells with a favorable therapeutic index, with enhanced killing of genomically high-risk CLL cells that are typically unresponsive to traditional therapies. More importantly, SINE slows disease progression, and improves overall survival in the Eµ-TCL1-SCID mouse model of CLL with minimal weight loss or other toxicities. Together, these findings demonstrate that XPO1 is a valid target in CLL with minimal effects on normal cells and provide a basis for the development of SINEs in CLL and related hematologic malignancies.
Subject(s)
Acrylates/pharmacology , Karyopherins/metabolism , Leukemia, Lymphocytic, Chronic, B-Cell/drug therapy , Receptors, Cytoplasmic and Nuclear/metabolism , Triazoles/pharmacology , Acrylates/chemistry , Acrylates/metabolism , Active Transport, Cell Nucleus/drug effects , Animals , Antineoplastic Agents/chemistry , Antineoplastic Agents/metabolism , Antineoplastic Agents/pharmacology , Cell Line, Tumor , Cell Survival/drug effects , Cells, Cultured , Crystallography, X-Ray , Humans , Immunoblotting , Interleukin-10/metabolism , Interleukin-6/metabolism , Karyopherins/chemistry , Karyopherins/genetics , Leukemia, Lymphocytic, Chronic, B-Cell/genetics , Leukemia, Lymphocytic, Chronic, B-Cell/metabolism , Mice , Mice, SCID , Mice, Transgenic , Microscopy, Confocal , Models, Molecular , Molecular Structure , Protein Binding , Protein Structure, Tertiary , Proto-Oncogene Proteins/genetics , Proto-Oncogene Proteins/metabolism , RNA Interference , Receptors, Cytoplasmic and Nuclear/chemistry , Receptors, Cytoplasmic and Nuclear/genetics , Reverse Transcriptase Polymerase Chain Reaction , T-Lymphocytes/drug effects , T-Lymphocytes/metabolism , Triazoles/chemistry , Triazoles/metabolism , Exportin 1 ProteinABSTRACT
Novel piperidine and piperazine derivatives have been designed and tested as inhibitors of LTA(4) hydrolase (LTA(4)H). Most potent compounds showed good potency in both enzymatic and functional human whole blood assay. Crystallography studies further confirmed observed structure-activity relationship and LTA(4)H binding mode for analogs from the piperidine series.
Subject(s)
Anti-Inflammatory Agents/chemistry , Enzyme Inhibitors/chemistry , Epoxide Hydrolases/antagonists & inhibitors , Piperazines/chemistry , Piperidines/chemistry , Anti-Inflammatory Agents/chemical synthesis , Anti-Inflammatory Agents/pharmacology , Binding Sites , Crystallography, X-Ray , Enzyme Inhibitors/chemical synthesis , Enzyme Inhibitors/pharmacology , Epoxide Hydrolases/metabolism , Humans , Piperazine , Piperazines/chemical synthesis , Piperazines/pharmacology , Piperidines/chemical synthesis , Piperidines/pharmacology , Structure-Activity RelationshipABSTRACT
Both in-house human genetic and literature data have converged on the identification of leukotriene 4 hydrolase (LTA(4)H) as a key target for the treatment of cardiovascular disease. We combined fragment-based crystallography screening with an iterative medicinal chemistry effort to optimize inhibitors of LTA(4)H. Ligand efficiency was followed throughout our structure-activity studies. As applied within the context of LTA(4)H inhibitor design, the chemistry team was able to design a potent compound 20 (DG-051) (K(d) = 26 nM) with high aqueous solubility (>30 mg/mL) and high oral bioavailability (>80% across species) that is currently undergoing clinical evaluation for the treatment of myocardial infarction and stroke. The structural biology-chemistry interaction described in this paper provides a sound alternative to conventional screening techniques. This is the first example of a gene-to-clinic paradigm enabled by a fragment-based drug discovery effort.
Subject(s)
Butyrates/pharmacology , Cardiovascular Diseases/drug therapy , Drug Discovery/methods , Epoxide Hydrolases/antagonists & inhibitors , Heterocyclic Compounds/pharmacology , Biological Availability , Butyrates/chemistry , Butyrates/therapeutic use , Crystallography, X-Ray , Enzyme Inhibitors/chemistry , Enzyme Inhibitors/therapeutic use , Epoxide Hydrolases/biosynthesis , Heterocyclic Compounds/chemistry , Heterocyclic Compounds/therapeutic use , Humans , Ligands , Myocardial Infarction/drug therapy , Peptide Fragments/chemistry , Solubility , Stroke/drug therapy , Structure-Activity RelationshipABSTRACT
A series of alpha-sulfone piperidine hydroxamate TACE inhibitors 11a-n bearing a quinolinyl methyl P1' group was prepared, and their activity was compared to analogous alpha- and beta-sulfone piperidine hydroxamates with a butynyloxy P1' group. The quinolinyl methyl P1' group affords increased inhibitory enzyme activity relative to the corresponding butynyloxy P1' analogs in the alpha-sulfone piperidine hydroxamate series, and greater selectivity than the corresponding butynyloxy P1' analogs in the beta-sulfone piperidine hydroxamate series.
Subject(s)
ADAM Proteins/antagonists & inhibitors , Hydroxamic Acids/chemical synthesis , Piperidines/chemistry , Protease Inhibitors/chemical synthesis , Sulfones/chemistry , ADAM Proteins/metabolism , ADAM17 Protein , Animals , Humans , Hydroxamic Acids/chemistry , Hydroxamic Acids/pharmacology , Mice , Piperidines/chemical synthesis , Protease Inhibitors/chemistry , Protease Inhibitors/pharmacology , Sulfones/chemical synthesisABSTRACT
Potent and selective TACE and MMP inhibitors utilizing the diazepine and thiazepine ring systems were synthesized and evaluated for biological activity in in vitro and in vivo models of TNF-alpha release. Oral activity in the mouse LPS model of TNF-alpha release was seen. Efficacy in the mouse collagen induced arthritis model was achieved with diazepine 20.
Subject(s)
Azepines/chemistry , Azepines/pharmacology , Matrix Metalloproteinase Inhibitors , Metalloendopeptidases/antagonists & inhibitors , ADAM Proteins , ADAM17 Protein , Animals , Azepines/chemical synthesis , Female , Mice , Mice, Inbred BALB C , Models, Chemical , Molecular Structure , Structure-Activity RelationshipABSTRACT
Class A-class C mechanism-based beta-lactamase inhibitors were designed on the basis of the intermediacy of an oxycarbenium species capable of cross-linking with amino acids residues in the active site. Penams 24 and 27 were very potent against AmpC in vitro. The MIC values of 24 in combination with piperacillin against class A and class C producing organisms showed improvement over clinically used tazobactam.
Subject(s)
Anti-Bacterial Agents/chemical synthesis , Bacterial Proteins , Cyclopropanes/chemical synthesis , Diazonium Compounds/chemistry , Enzyme Inhibitors/chemical synthesis , Rhodium , Serine/chemistry , Spiro Compounds/chemical synthesis , Sulbactam/chemistry , beta-Lactamase Inhibitors , beta-Lactams/chemical synthesis , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/pharmacology , Catalysis , Crystallography, X-Ray , Cyclopropanes/chemistry , Cyclopropanes/pharmacology , Enzyme Inhibitors/chemistry , Enzyme Inhibitors/pharmacology , Escherichia coli/drug effects , Microbial Sensitivity Tests , Models, Molecular , Pseudomonas aeruginosa/drug effects , Serratia marcescens/drug effects , Spiro Compounds/chemistry , Spiro Compounds/pharmacology , Structure-Activity Relationship , Sulbactam/analogs & derivatives , beta-Lactamases/chemistry , beta-Lactams/chemistry , beta-Lactams/pharmacologyABSTRACT
The matrix metalloproteinases (MMPs) are a family of zinc-containing endopeptidases that play a key role in both physiological and pathological tissue degradation. In our preceding paper, we have reported on a series of novel and orally active N-hydroxy-alpha-phenylsulfonylacetamide derivatives. However, these compounds had two drawbacks (moderate selectivity and chirality issues). To circumvent these two problems, a series of novel and orally active N-substituted 4-benzenesulfonylpiperidine-4-carboxylic acid hydroxyamide derivatives have been synthesized. The present paper deals with the synthesis and SAR of these compounds. Among the several compounds synthesized, derivative 55 turned out to be a potent, selective, and an orally active MMP inhibitor in the clinically relevant advanced rabbit osteoarthritis model. Detailed pharmacokinetics and metabolism data are described.
Subject(s)
Hydroxamic Acids/chemical synthesis , Matrix Metalloproteinase Inhibitors , Osteoarthritis/drug therapy , Piperidines/chemical synthesis , Protease Inhibitors/chemical synthesis , Sulfones/chemical synthesis , ADAM Proteins , ADAM17 Protein , Administration, Oral , Animals , Binding Sites , Biological Assay , Cartilage/drug effects , Cartilage/enzymology , Cattle , Crystallography, X-Ray , Dialysis , Dogs , Haplorhini , Humans , Hydroxamic Acids/pharmacokinetics , Hydroxamic Acids/pharmacology , Male , Matrix Metalloproteinase 13 , Matrix Metalloproteinases/chemistry , Metalloendopeptidases/antagonists & inhibitors , Mice , Models, Molecular , Piperidines/pharmacokinetics , Piperidines/pharmacology , Protease Inhibitors/pharmacokinetics , Protease Inhibitors/pharmacology , Rabbits , Rats , Structure-Activity Relationship , Sulfones/pharmacokinetics , Sulfones/pharmacologyABSTRACT
Resistance to antibiotics is currently a major health concern in treating infectious diseases. The most common mechanism of resistance to beta-lactam antibiotics is the production of beta-lactamases, which destroy beta-lactam antibiotics before they reach the bacterial target. Combination therapy, which involves treatment with a beta-lactam antibiotic and a beta-lactamase inhibitor, has been successfully used to control resistance during last two decades. Due to the lack of effectiveness of the currently available beta-lactamase inhibitors against class C enzymes and new variants of beta-lactamases, there is a need to develop an inhibitor with broad-spectrum activity. Since the discovery of clavulanic acid, there has been an enormous research effort in this area to identify better antibiotic/inhibitor combinations and to understand the molecular bases for interactions between beta-lactam antibiotics, beta-lactamases, and beta-lactamase inhibitors. This review describes some of the structure- and mechanism-based approaches to design of new beta-lactamase inhibitors and the study of probable mechanisms of inhibition using X-ray, electrospray ionization mass spectrometry, and molecular modeling techniques.
