ABSTRACT
RORγ plays a critical role in controlling a pro-inflammatory gene expression program in several lymphocyte lineages including T cells, γδ T cells, and innate lymphoid cells. RORγ-mediated inflammation has been linked to susceptibility to Crohn's disease, arthritis, and psoriasis. Thus inverse agonists of RORγ have the potential of modulating inflammation. Our goal was to optimize two RORγ inverse agonists: T0901317 from literature and 1 that we obtained from internal screening. We used information from internal X-ray structures to design two libraries that led to a new biaryl series.
Subject(s)
Hydrocarbons, Fluorinated/chemistry , Nuclear Receptor Subfamily 1, Group F, Member 3/agonists , Structure-Activity Relationship , Sulfonamides/chemistry , Binding Sites , Crystallography, X-Ray , Drug Design , Hydrocarbons, Fluorinated/pharmacology , Models, Molecular , Molecular Docking Simulation , Molecular Structure , Nuclear Receptor Subfamily 1, Group F, Member 3/chemistry , Nuclear Receptor Subfamily 1, Group F, Member 3/metabolism , Sulfonamides/pharmacologyABSTRACT
The nuclear receptor RORγ plays a central role in controlling a pro-inflammatory gene expression program in several lymphocyte lineages including TH17 cells. RORγ-dependent inflammation has been implicated in the pathogenesis of several major autoimmune diseases and thus RORγ is an attractive target for therapeutic intervention in these diseases. Starting from a lead biaryl compound 4a, replacement of the head phenyl moiety with a substituted aminopyrazole group resulted in a series with improved physical properties. Further SAR exploration led to analogues (e.g., 4j and 5m) as potent RORγ inverse agonists.
Subject(s)
Benzamides/chemistry , Benzamides/pharmacology , Drug Inverse Agonism , Nuclear Receptor Subfamily 1, Group F, Member 3/antagonists & inhibitors , Pyrazoles/chemistry , Pyrazoles/pharmacology , Animals , Autoimmune Diseases/drug therapy , Autoimmune Diseases/immunology , Humans , Interleukin-17/immunology , Mice , Models, Molecular , Nuclear Receptor Subfamily 1, Group F, Member 3/chemistry , Nuclear Receptor Subfamily 1, Group F, Member 3/immunology , Spleen/cytology , Spleen/drug effects , Spleen/immunology , Th17 Cells/drug effects , Th17 Cells/immunologyABSTRACT
RORγt is a pivotal regulator of a pro-inflammatory gene expression program implicated in the pathology of several major human immune-mediated diseases. Evidence from mouse models demonstrates that genetic or pharmacological inhibition of RORγ activity can block the production of pathogenic cytokines, including IL-17, and convey therapeutic benefit. We have identified and developed a biaryl-carboxylamide series of RORγ inverse agonists via a structure based design approach. Co-crystal structures of compounds 16 and 48 supported the design approach and confirmed the key interactions with RORγ protein; the hydrogen bonding with His479 was key to the significant improvement in inverse agonist effect. The results have shown this is a class of potent and selective RORγ inverse agonists, with demonstrated oral bioavailability in rodents.
Subject(s)
Amides/chemistry , Amides/pharmacology , Biphenyl Compounds/chemistry , Biphenyl Compounds/pharmacology , Drug Inverse Agonism , Nuclear Receptor Subfamily 1, Group F, Member 3/agonists , Nuclear Receptor Subfamily 1, Group F, Member 3/antagonists & inhibitors , Amides/pharmacokinetics , Animals , Biphenyl Compounds/pharmacokinetics , Cell Line , Cytokines/immunology , Drug Discovery , Humans , Hydrogen Bonding , Interleukin-17/immunology , Mice , Molecular Docking Simulation , Nuclear Receptor Subfamily 1, Group F, Member 3/immunology , RatsABSTRACT
The metalloprotease ADAMTS-5 is considered a potential target for the treatment of osteoarthritis. To identify selective inhibitors of ADAMTS-5, we employed encoded library technology (ELT), which enables affinity selection of small molecule binders from complex mixtures by DNA tagging. Selection of ADAMTS-5 against a four-billion member ELT library led to a novel inhibitor scaffold not containing a classical zinc-binding functionality. One exemplar, (R)-N-((1-(4-(but-3-en-1-ylamino)-6-(((2-(thiophen-2-yl)thiazol-4-yl)methyl)amino)-1,3,5-triazin-2-yl)pyrrolidin-2-yl)methyl)-4-propylbenzenesulfonamide (8), inhibited ADAMTS-5 with IC(50) = 30 nM, showing >50-fold selectivity against ADAMTS-4 and >1000-fold selectivity against ADAMTS-1, ADAMTS-13, MMP-13, and TACE. Extensive SAR studies showed that potency and physicochemical properties of the scaffold could be further improved. Furthermore, in a human osteoarthritis cartilage explant study, compounds 8 and 15f inhibited aggrecanase-mediated (374)ARGS neoepitope release from aggrecan and glycosaminoglycan in response to IL-1ß/OSM stimulation. This study provides the first small molecule evidence for the critical role of ADAMTS-5 in human cartilage degradation.
Subject(s)
ADAM Proteins/antagonists & inhibitors , Cartilage, Articular/drug effects , Databases, Chemical , Osteoarthritis/pathology , Sulfonamides/chemical synthesis , Triazines/chemical synthesis , ADAMTS5 Protein , Aggrecans/metabolism , Animals , Cartilage, Articular/metabolism , Cartilage, Articular/pathology , Endopeptidases/metabolism , Epitopes , Glycosaminoglycans/metabolism , Humans , In Vitro Techniques , Male , Middle Aged , Osteoarthritis/drug therapy , Rats , Rats, Sprague-Dawley , Small Molecule Libraries , Stereoisomerism , Structure-Activity Relationship , Sulfonamides/pharmacokinetics , Sulfonamides/pharmacology , Triazines/pharmacokinetics , Triazines/pharmacologyABSTRACT
Biochemical combinatorial techniques such as phage display, RNA display and oligonucleotide aptamers have proven to be reliable methods for generation of ligands to protein targets. Adapting these techniques to small synthetic molecules has been a long-sought goal. We report the synthesis and interrogation of an 800-million-member DNA-encoded library in which small molecules are covalently attached to an encoding oligonucleotide. The library was assembled by a combination of chemical and enzymatic synthesis, and interrogated by affinity selection. We describe methods for the selection and deconvolution of the chemical display library, and the discovery of inhibitors for two enzymes: Aurora A kinase and p38 MAP kinase.
Subject(s)
DNA/chemistry , Drug Design , Protein Kinase Inhibitors/chemical synthesis , Small Molecule Libraries/chemical synthesis , Animals , Aurora Kinases , Combinatorial Chemistry Techniques , DNA/genetics , Models, Molecular , Protein Kinase Inhibitors/chemistry , Protein Kinase Inhibitors/pharmacology , Protein Serine-Threonine Kinases/antagonists & inhibitors , Small Molecule Libraries/chemistry , Small Molecule Libraries/pharmacology , p38 Mitogen-Activated Protein Kinases/antagonists & inhibitorsABSTRACT
Potent and selective antagonists of the adenosine A2A receptor often contain a nitrogen-rich fused-ring heterocyclic core. Replacement of the core with an isomeric ring system has previously been shown to improve target affinity, selectivity, and in vivo activity. This paper describes the preparation, by a novel route, of A2A receptor antagonists containing the [1,2,4]triazolo[1,5-a]pyrazine nucleus, which is isomeric with the [1,2,4]triazolo[1,5-c]pyrimidine core of a series of known A2A antagonists with in vivo activity in animal models of Parkinson's disease.
