ABSTRACT
The nuclear hormone receptor retinoic acid receptor-related orphan C2 (RORC2, also known as RORγt) is a promising target for the treatment of autoimmune diseases. A small molecule, inverse agonist of the receptor is anticipated to reduce production of IL-17, a key proinflammatory cytokine. Through a high-throughput screening approach, we identified a molecule displaying promising binding affinity for RORC2, inhibition of IL-17 production in Th17 cells, and selectivity against the related RORA and RORB receptor isoforms. Lead optimization to improve the potency and metabolic stability of this hit focused on two key design strategies, namely, iterative optimization driven by increasing lipophilic efficiency and structure-guided conformational restriction to achieve optimal ground state energetics and maximize receptor residence time. This approach successfully identified 3-cyano- N-(3-(1-isobutyrylpiperidin-4-yl)-1-methyl-4-(trifluoromethyl)-1 H-pyrrolo[2,3- b]pyridin-5-yl)benzamide as a potent and selective RORC2 inverse agonist, demonstrating good metabolic stability, oral bioavailability, and the ability to reduce IL-17 levels and skin inflammation in a preclinical in vivo animal model upon oral administration.
Subject(s)
Drug Design , Drug Inverse Agonism , Nuclear Receptor Subfamily 1, Group F, Member 3/agonists , Pyridines/administration & dosage , Pyridines/pharmacology , Administration, Oral , Animals , Biological Availability , Drug Evaluation, Preclinical , Humans , Mice , Pyridines/pharmacokinetics , Th17 Cells/drug effects , Th17 Cells/metabolismABSTRACT
Fatty acid amide hydrolase (FAAH) is an integral membrane serine hydrolase responsible for the degradation of fatty acid amide signaling molecules such as endocannabinoid anandamide (AEA), which has been shown to possess cannabinoid-like analgesic properties. Herein we report the optimization of spirocyclic 7-azaspiro[3.5]nonane and 1-oxa-8-azaspiro[4.5]decane urea covalent inhibitors of FAAH. Using an iterative design and optimization strategy, lead compounds were identified with a remarkable reduction in molecular weight and favorable CNS drug like properties. 3,4-Dimethylisoxazole and 1-methyltetrazole were identified as superior urea moieties for this inhibitor class. A dual purpose in vivo efficacy and pharmacokinetic screen was designed to be the key decision enabling experiment affording the ability to move quickly from compound synthesis to selection of preclinical candidates. On the basis of the remarkable potency, selectivity, pharmacokinetic properties and in vivo efficacy, PF-04862853 (15p) was advanced as a clinical candidate.
Subject(s)
Amidohydrolases/antagonists & inhibitors , Analgesics/pharmacology , Drug Discovery , Enzyme Inhibitors/pharmacology , Pain/drug therapy , Spiro Compounds/pharmacology , Administration, Oral , Analgesics/administration & dosage , Analgesics/chemistry , Analgesics/therapeutic use , Animals , Aza Compounds/administration & dosage , Aza Compounds/chemistry , Aza Compounds/pharmacology , Aza Compounds/therapeutic use , Biological Availability , Chromatography, High Pressure Liquid , Drug Evaluation, Preclinical , Enzyme Inhibitors/administration & dosage , Enzyme Inhibitors/chemistry , Enzyme Inhibitors/therapeutic use , Rats , Spiro Compounds/administration & dosage , Spiro Compounds/chemistry , Spiro Compounds/therapeutic useABSTRACT
Herein we report the identification of two new fatty acid amide hydrolase (FAAH) inhibitor lead series with FAAH k(inact)/K(i) potency values greater than 1500M(-1)s(-1). The two novel spirocyclic cores, 7-azaspiro[3.5]nonane and 1-oxa-8-azaspiro[4.5]decane, clearly distinguished themselves from the other spirocyclic cores on the basis of their superior potency for FAAH. Lead compounds from these two series have suitable FAAH potency and selectivity for additional medicinal chemistry optimization.
Subject(s)
Amidohydrolases/antagonists & inhibitors , Aza Compounds/pharmacology , Drug Discovery , Enzyme Inhibitors/pharmacology , Spiro Compounds/pharmacology , Aza Compounds/chemistry , Enzyme Inhibitors/chemistry , Models, Molecular , Spiro Compounds/chemistry , Structure-Activity RelationshipABSTRACT
We have discovered a novel class of nonsteroidal pyrazoline antagonists of the mineralocorticoid receptor (MR) that show excellent potency and selectivity against other nuclear receptors. Early analogues were poorly soluble and had a propensity to inhibit the hERG channel. Remarkably, both of these challenges were overcome by incorporation of a single carboxylate moiety. Structural modification of carboxylate-containing lead R-4g with a wide range of substituents at each position of the pyrazoline ring resulted in R-12o, which shows excellent activity against MR and reasonable pharmacokinetic profile. Introduction of conformational restriction led to a novel series characterized by exquisite potency and favorable steroid receptor selectivity and pharmacokinetic profile. Oral dosing of 3S,3aR-27d (PF-3882845) in the Dahl salt sensitive preclinical model of salt-induced hypertension and nephropathy showed blood pressure attenuation significantly greater than that with eplerenone, reduction in urinary albumin, and renal protection. As a result of these findings, 3S,3aR-27d was advanced to clinical studies.
Subject(s)
Antihypertensive Agents/chemical synthesis , Hypertension/drug therapy , Indazoles/chemical synthesis , Kidney Diseases/drug therapy , Mineralocorticoid Receptor Antagonists , Nitriles/chemical synthesis , Animals , Antihypertensive Agents/pharmacokinetics , Antihypertensive Agents/pharmacology , Blood Pressure/drug effects , Cell Line, Tumor , Chlorobenzenes , Crystallography, X-Ray , Humans , Indazoles/pharmacokinetics , Indazoles/pharmacology , Indenes , Male , Models, Molecular , Molecular Conformation , Nitriles/pharmacokinetics , Nitriles/pharmacology , Radioligand Assay , Rats , Rats, Inbred Dahl , Rats, Sprague-Dawley , Stereoisomerism , Structure-Activity RelationshipABSTRACT
The work described herein demonstrates the utility of structure-based drug design (SBDD) in shifting the binding mode of an HTS hit from a DFG-in to a DFG-out binding mode resulting in a class of novel potent CSF-1R kinase inhibitors suitable for lead development.
Subject(s)
Protein Kinase Inhibitors/chemistry , Receptor, Macrophage Colony-Stimulating Factor/antagonists & inhibitors , Binding Sites , Crystallography, X-Ray , Drug Design , High-Throughput Screening Assays , Hydrogen Bonding , Molecular Conformation , Protein Binding , Protein Kinase Inhibitors/chemical synthesis , Protein Kinase Inhibitors/pharmacology , Receptor, Macrophage Colony-Stimulating Factor/metabolismABSTRACT
Optimization of kinase selectivity for a set of benzothiophene MK2 inhibitors provided analogs with potencies of less than 500 nM in a cell based assay. The selectivity of the inhibitors can be rationalized by examination of X-ray crystal structures of inhibitors bound to MK2.
Subject(s)
MAP Kinase Kinase 2/antagonists & inhibitors , Protein Kinase Inhibitors/chemistry , Thiophenes/chemistry , Binding Sites , Cell Line, Tumor , Crystallography, X-Ray , Cyclin-Dependent Kinase 2/antagonists & inhibitors , Cyclin-Dependent Kinase 2/metabolism , Drug Design , Humans , MAP Kinase Kinase 2/metabolism , Protein Kinase Inhibitors/chemical synthesis , Protein Kinase Inhibitors/pharmacology , Thiophenes/chemical synthesis , Thiophenes/pharmacologyABSTRACT
Identification of potent benzothiophene inhibitors of mitogen activated protein kinase-activated protein kinase 2 (MK2), structure-activity relationship (SAR) studies, selectivity assessments against CDK2, cellular potency and mechanism of action are presented. Crystallographic data provide a rationale for the observed MK2 potency as well as selectivity over CDK2 for this class of inhibitors.
