ABSTRACT
NF-κB-inducing kinase (NIK; also known as MAP3K14) is a central regulator of non-canonical NF-κB signaling in response to stimulation of TNF receptor superfamily members, such as the lymphotoxin-ß receptor (LTßR), and is implicated in pathological angiogenesis associated with chronic inflammation and cancer. Here, we identify a previously unrecognized role of the LTßR-NIK axis during inflammatory activation of human endothelial cells (ECs). Engagement of LTßR-triggered canonical and non-canonical NF-κB signaling promoted expression of inflammatory mediators and adhesion molecules, and increased immune cell adhesion to ECs. Sustained LTßR-induced inflammatory activation of ECs was NIK dependent, but independent of p100, indicating that the non-canonical arm of NF-κB is not involved. Instead, prolonged activation of canonical NF-κB signaling, through the interaction of NIK with IκB kinase α and ß (also known as CHUK and IKBKB, respectively), was required for the inflammatory response. Endothelial inflammatory activation induced by synovial fluid from rheumatoid arthritis patients was significantly reduced by NIK knockdown, suggesting that NIK-mediated alternative activation of canonical NF-κB signaling is a key driver of pathological inflammatory activation of ECs. Targeting NIK could thus provide a novel approach for treating chronic inflammatory diseases.
Subject(s)
Endothelial Cells/metabolism , Lymphotoxin beta Receptor/metabolism , NF-kappa B/metabolism , Protein Serine-Threonine Kinases/metabolism , Signal Transduction , Basic Helix-Loop-Helix Transcription Factors , Cell Line , Cells, Cultured , Endothelium/metabolism , Gene Expression Regulation , Humans , NF-kappa B/genetics , Neovascularization, Pathologic/metabolism , Protein Serine-Threonine Kinases/genetics , NF-kappaB-Inducing KinaseABSTRACT
One of the youngest people to ever qualify as a vet, he was a life-long practitioner whose philosophy was one of caring - whether it be for animals, student vets, farmers, local people or friends.
ABSTRACT
The cysteine protease cruzipain is considered to be a validated target for therapeutic intervention in the treatment of Chagas disease. Anti-trypanosomal activity against the CL Brener strain of T. cruzi was observed in the 0.1 µM to 1 µM range for three nitrile-based cysteine protease inhibitors based on two scaffolds known to be associated with cathepsin K inhibition. The two compounds showing the greatest potency against the trypanosome were characterized by EC50 values (0.12 µM and 0.25 µM) that were an order of magnitude lower than the corresponding Ki values measured against cruzain, a recombinant form of cruzipain, in an enzyme inhibition assay. This implies that the anti-trypanosomal activity of these two compounds may not be explained only by the inhibition of the cruzain enzyme, thereby triggering a putative polypharmacological profile towards cysteine proteases.
Subject(s)
Cysteine Proteinase Inhibitors/pharmacology , Nitriles/pharmacology , Trypanocidal Agents/pharmacology , Cysteine Endopeptidases , Cysteine Proteinase Inhibitors/chemical synthesis , Cysteine Proteinase Inhibitors/chemistry , Models, Molecular , Molecular Structure , Nitriles/chemical synthesis , Nitriles/chemistry , Parasitic Sensitivity Tests , Protozoan Proteins/antagonists & inhibitors , Trypanocidal Agents/chemical synthesis , Trypanocidal Agents/chemistry , Trypanosoma cruzi/drug effectsABSTRACT
A lead generation and optimization program delivered the highly selective and potent CatC inhibitor 10 as an in vivo tool compound and potential development candidate. Structural studies were undertaken to generate SAR understanding.
Subject(s)
Cathepsin C/antagonists & inhibitors , Protease Inhibitors/chemical synthesis , Protease Inhibitors/pharmacology , Humans , Indicators and Reagents , Models, Molecular , Molecular Conformation , Recombinant Proteins/chemistry , Spectrometry, Fluorescence , Structure-Activity Relationship , Substrate Specificity , X-Ray DiffractionABSTRACT
The discovery of potent small molecule dual antagonists of the human CCR3 and H(1) receptors is described for the treatment of allergic diseases, for example, asthma and allergic rhinitis. Optimizing in vitro potency and metabolic stability, starting from a CCR1 lead compound, led to compound 20 with potent dual CCR3/H(1) activity and in vitro metabolic stability.
Subject(s)
Drug Discovery , Hydroxamic Acids/pharmacology , Piperidines/pharmacology , Receptors, CCR3/antagonists & inhibitors , Receptors, Histamine H1/metabolism , Animals , Hepatocytes/chemistry , Hepatocytes/metabolism , Humans , Hydroxamic Acids/chemistry , Hydroxamic Acids/metabolism , Microsomes, Liver/chemistry , Microsomes, Liver/metabolism , Piperidines/chemistry , Piperidines/metabolism , Rats , Structure-Activity Relationship , Tissue DistributionABSTRACT
The second part of this communication focuses on the resolution of issues surrounding the series of hydroxyamide phenoxypiperidine CCR3/H(1) dual antagonists described in Part I. This involved further structural exploration directed at reducing metabolism and leading to the identification of compound 60 with a greatly improved in vivo pharmacokinetic profile.
Subject(s)
Drug Discovery , Piperidines/pharmacology , Receptors, CCR3/antagonists & inhibitors , Receptors, Histamine H1/metabolism , Animals , Dogs , Hepatocytes/chemistry , Hepatocytes/metabolism , Humans , Microsomes, Liver/chemistry , Microsomes, Liver/metabolism , Piperidines/chemistry , Piperidines/metabolism , Rats , Structure-Activity Relationship , Tissue DistributionABSTRACT
Enantioselective multicomponent reaction: in the presence of a catalytic amount of chiral BINOL-derived phosphoric acid (TRIP), the reaction of an α-isocyanoacetate 1, an aldehyde 2, and an aniline 3, followed by addition of a toluene solution of α,ß-unsaturated acyl chloride 4 afforded the oxa-bridged tricycle 5 in excellent yield, diastereoselectivity, and enantioselectivity. Six chemical bonds, five stereogenic centers, and three cycles were formed in this one-pot four-component reaction.
Subject(s)
Matrix Metalloproteinase Inhibitors , Protease Inhibitors/chemistry , Amino Acid Sequence , Binding Sites , Catalytic Domain , Computer Simulation , Crystallography, X-Ray , Drug Evaluation, Preclinical , High-Throughput Screening Assays , Matrix Metalloproteinase 7/metabolism , Molecular Sequence Data , Protease Inhibitors/chemical synthesis , Protease Inhibitors/pharmacology , Sequence AlignmentABSTRACT
We describe the discovery of small molecule benzazepine derivatives as agonists of human peroxisome proliferator-activated receptor δ (PPARδ) that displayed excellent selectivity over the PPARα and PPARγ subtypes. Compound 8 displayed good PK in the rat and efficacy in upregulation of pyruvate dehydrogenase kinase, isozyme 4 (PDK4) mRNA in human primary myotubes, a biomarker for increased fatty acid oxidation.
Subject(s)
Anilides/chemical synthesis , Benzazepines/chemistry , PPAR delta/agonists , Anilides/chemistry , Anilides/pharmacokinetics , Animals , Benzazepines/chemical synthesis , Benzazepines/pharmacokinetics , Binding Sites , Computer Simulation , Hepatocytes/metabolism , Humans , Microsomes, Liver/metabolism , PPAR alpha/agonists , PPAR alpha/metabolism , PPAR delta/metabolism , PPAR gamma/agonists , PPAR gamma/metabolism , Protein Serine-Threonine Kinases/genetics , Protein Serine-Threonine Kinases/metabolism , Pyruvate Dehydrogenase Acetyl-Transferring Kinase , Rats , Up-RegulationABSTRACT
Small molecule isoindoline and tetrahydroisoquinoline derivatives have been identified as selective agonists of human peroxisome proliferator-activated receptor δ (PPARδ. Compound 18 demonstrated efficacy in a biomarker for increased fatty acid oxidation, with upregulation of pyruvate dehydrogenase kinase, isozyme 4 (PDK4) in human primary myotubes.
Subject(s)
Indoles/chemistry , PPAR delta/agonists , Tetrahydroisoquinolines/chemistry , Binding Sites , Catalytic Domain , Computer Simulation , Humans , Indoles/chemical synthesis , Indoles/pharmacology , PPAR delta/metabolism , Protein Serine-Threonine Kinases/genetics , Protein Serine-Threonine Kinases/metabolism , Pyruvate Dehydrogenase Acetyl-Transferring Kinase , Structure-Activity Relationship , Tetrahydroisoquinolines/chemical synthesis , Tetrahydroisoquinolines/pharmacologyABSTRACT
The P2X7 receptor (P2X7R) has been implicated in the process of multinucleation and cell fusion. We have previously demonstrated that blockade of P2X7Rs on osteoclast precursors using a blocking antibody inhibited multinucleated osteoclast formation in vitro, but that P2X7R KO mice maintain the ability to form multinucleated osteoclasts. This apparent contradiction of the role the P2X7R plays in multinucleation has prompted us to examine the effect of the most commonly used and recently available P2X7R antagonists on osteoclast formation and function. When added to recombinant RANKL and M-CSF human blood monocytes cultures, all but one compound, decreased the formation and function of multinucleated TRAP-positive osteoclasts in a concentration-dependent manner. These data provide further evidence for the role of the P2X7R in the formation of functional human multinucleated osteoclasts and highlight the importance of selection of antagonists for use in long-term experiments.
Subject(s)
Drug Discovery , Purinergic P2 Receptor Antagonists , Amides/chemistry , Amides/pharmacology , Amides/therapeutic use , Amidines/chemistry , Amidines/pharmacology , Amidines/therapeutic use , Animals , Arthritis, Rheumatoid/drug therapy , Azoles/chemistry , Azoles/pharmacology , Azoles/therapeutic use , Humans , Receptors, Purinergic P2X7 , Structure-Activity RelationshipABSTRACT
A novel series of antagonists of the human P2X7 receptor is described. Modification of substituents enabled identification of compounds selective for the rat P2X7 receptor and provides useful pharmacological tools for evaluation of the role of P2X7 in disease.
Subject(s)
Adamantane/analogs & derivatives , Adamantane/chemical synthesis , Benzamides/chemical synthesis , Interleukin-1beta/antagonists & inhibitors , Purinergic P2 Receptor Antagonists , Adamantane/pharmacology , Animals , Benzamides/pharmacology , Blood Proteins/metabolism , Crystallography, X-Ray , Humans , In Vitro Techniques , Molecular Conformation , Monocytes/metabolism , Protein Binding , Rats , Receptors, Purinergic P2X7 , Structure-Activity RelationshipABSTRACT
Palladium catalysed selective double insertion of isonitriles into aryl bromides with trapping by sodium alkoxides provides an efficient 4-component synthesis of unusual alpha-iminoimidates.