ABSTRACT
BACKGROUND: Inflammation in the central nervous system (CNS) is observed in many neurological disorders. Nitric oxide-soluble guanylate cyclase-cyclic guanosine monophosphate (NO-sGC-cGMP) signaling plays an essential role in modulating neuroinflammation. CYR119 is a CNS-penetrant sGC stimulator that amplifies endogenous NO-sGC-cGMP signaling. We evaluated target engagement and the effects of CYR119 on markers of neuroinflammation in vitro in mouse microglial cells and in vivo in quinolinic acid (QA)-induced and high-fat diet-induced rodent neuroinflammation models. METHODS: Target engagement was verified in human embryonic kidney (HEK) cells, rat primary neurons, mouse SIM-A9 cells, and in rats by measuring changes in cGMP and downstream targets of sGC signaling [phosphorylated vasodilator-stimulated phosphoprotein (pVASP), phosphorylated cAMP-response element binding (pCREB)]. In SIM-A9 cells stimulated with lipopolysaccharides (LPS), markers of inflammation were measured when cells were treated with or without CYR119. In rats, microinjections of QA and vehicle were administered into the right and left hemispheres of striatum, respectively, and then rats were dosed daily with either CYR119 (10 mg/kg) or vehicle for 7 days. The activation of microglia [ionized calcium binding adaptor molecule 1 (Iba1)] and astrocytes [glial fibrillary acidic protein (GFAP)] was measured by immunohistochemistry. Diet-induced obese (DIO) mice were treated daily with CYR119 (10 mg/kg) for 6 weeks, after which inflammatory genetic markers were analyzed in the prefrontal cortex. RESULTS: In vitro, CYR119 synergized with exogenous NO to increase the production of cGMP in HEK cells and in primary rat neuronal cell cultures. In primary neurons, CYR119 stimulated sGC, resulting in accumulation of cGMP and phosphorylation of CREB, likely through the activation of protein kinase G (PKG). CYR119 attenuated LPS-induced elevation of interleukin 6 (IL-6) and tumor necrosis factor (TNF) in mouse microglial cells. Following oral dosing in rats, CYR119 crossed the blood-brain barrier (BBB) and stimulated an increase in cGMP levels in the cerebral spinal fluid (CSF). In addition, levels of proinflammatory markers associated with QA administration or high-fat diet feeding were lower in rodents treated with CYR119 than in those treated with vehicle. CONCLUSIONS: These data suggest that sGC stimulation could provide neuroprotective effects by attenuating inflammatory responses in nonclinical models of neuroinflammation.
Subject(s)
Anti-Inflammatory Agents/metabolism , Central Nervous System/metabolism , Cyclic GMP/metabolism , Inflammation Mediators/metabolism , Neurons/metabolism , Soluble Guanylyl Cyclase/metabolism , Animals , Anti-Inflammatory Agents/pharmacology , Biomarkers/metabolism , Cells, Cultured , Central Nervous System/drug effects , Dose-Response Relationship, Drug , HEK293 Cells , Humans , Inflammation Mediators/antagonists & inhibitors , Male , Mice , Mice, Inbred C57BL , Neurons/drug effects , Rats , Rats, Sprague-DawleyABSTRACT
Effective treatments for neurodegenerative diseases remain elusive and are critically needed since the burden of these diseases increases across an aging global population. Nitric oxide (NO) is a gasotransmitter that binds to soluble guanylate cyclase (sGC) to produce cyclic guanosine monophosphate (cGMP). Impairment of this pathway has been demonstrated in neurodegenerative diseases. Normalizing deficient NO-cGMP signaling could address multiple pathophysiological features of neurodegenerative diseases. sGC stimulators are small molecules that synergize with NO, activate sGC, and increase cGMP production. Many systemic sGC stimulators have been characterized and advanced into clinical development for a variety of non-central nervous system (CNS) pathologies. Here, we disclose the discovery of CY6463, the first brain-penetrant sGC stimulator in clinical development for the treatment of neurodegenerative diseases, and demonstrate its ability to improve neuronal activity, mediate neuroprotection, and increase cognitive performance in preclinical models. In several cellular assays, CY6463 was demonstrated to be a potent stimulator of sGC. In agreement with the known effects of sGC stimulation in the vasculature, CY6463 elicits decreases in blood pressure in both rats and mice. Relative to a non-CNS penetrant sGC stimulator, rodents treated with CY6463 had higher cGMP levels in cerebrospinal fluid (CSF), functional-magnetic-resonance-imaging-blood-oxygen-level-dependent (fMRI-BOLD) signals, and cortical electroencephalographic (EEG) gamma-band oscillatory power. Additionally, CY6463 improved cognitive performance in a model of cognitive disruption induced by the administration of a noncompetitive N-methyl-D-aspartate (NMDA) receptor antagonist. In models of neurodegeneration, CY6463 treatment increased long-term potentiation (LTP) in hippocampal slices from a Huntington's disease mouse model and decreased the loss of dendritic spines in aged and Alzheimer's disease mouse models. In a model of diet-induced obesity, CY6463 reduced markers of inflammation in the plasma. Furthermore, CY6463 elicited an additive increase in cortical gamma-band oscillatory power when co-administered with donepezil: the standard of care in Alzheimer's disease. Together, these data support the clinical development of CY6463 as a novel treatment for neurodegenerative disorders.
ABSTRACT
Soluble guanylate cyclase (sGC) is a clinically validated therapeutic target in the treatment of pulmonary hypertension. Modulators of sGC have the potential to treat diseases that are affected by dysregulation of the NO-sGC-cGMP signal transduction pathway. This letter describes the SAR efforts that led to the discovery of CYR715, a novel carboxylic acid-containing sGC stimulator, with an improved metabolic profile relative to our previously described stimulator, IWP-051. CYR715 addressed potential idiosyncratic drug toxicity (IDT) liabilities associated with the formation of reactive, migrating acyl glucuronides (AG) found in related carboxylic acid-containing analogs and demonstrated high oral bioavailability in rat and dose-dependent hemodynamic pharmacology in normotensive Sprague-Dawley rats.
Subject(s)
Carboxylic Acids/chemistry , Glucuronides/chemistry , Hypertension, Pulmonary/drug therapy , Soluble Guanylyl Cyclase/metabolism , Vasodilator Agents/chemistry , Administration, Oral , Animals , Dose-Response Relationship, Drug , Drug Evaluation, Preclinical , Glucuronides/administration & dosage , Glucuronides/pharmacokinetics , Humans , Male , Metabolome , Models, Molecular , Nitric Oxide/metabolism , Oxidative Stress/drug effects , Protein Binding , Rats, Sprague-Dawley , Signal Transduction , Structure-Activity Relationship , Vasodilator Agents/administration & dosage , Vasodilator Agents/pharmacokineticsABSTRACT
A highly potent and selective DGAT-1 inhibitor was identified and used in rodent models of obesity and postprandial chylomicron excursion to validate DGAT-1 inhibition as a novel approach for the treatment of metabolic diseases. Specifically, compound 4a conferred weight loss and a reduction in liver triglycerides when dosed chronically in DIO mice and depleted serum triglycerides following a lipid challenge in a dose-dependent manner, thus, reproducing major phenotypical characteristics of DGAT-1(-/-) mice.
Subject(s)
Anti-Obesity Agents/chemical synthesis , Cycloheptanes/chemical synthesis , Diacylglycerol O-Acyltransferase/antagonists & inhibitors , Hypolipidemic Agents/chemical synthesis , Keto Acids/chemical synthesis , Urea/analogs & derivatives , Urea/chemical synthesis , Animals , Anti-Obesity Agents/pharmacokinetics , Anti-Obesity Agents/pharmacology , Biphenyl Compounds/chemical synthesis , Biphenyl Compounds/pharmacokinetics , Biphenyl Compounds/pharmacology , Cycloheptanes/pharmacokinetics , Cycloheptanes/pharmacology , Diacylglycerol O-Acyltransferase/genetics , Eating/drug effects , Humans , Hypolipidemic Agents/pharmacokinetics , Hypolipidemic Agents/pharmacology , Isoenzymes/antagonists & inhibitors , Isoenzymes/genetics , Keto Acids/pharmacokinetics , Keto Acids/pharmacology , Liver/metabolism , Mice , Mice, Mutant Strains , Stereoisomerism , Structure-Activity Relationship , Triglycerides/metabolism , Urea/pharmacokinetics , Urea/pharmacology , Weight LossABSTRACT
AMP-activated protein kinase (AMPK) is well established as a sensor and regulator of intracellular and whole-body energy metabolism. A high-throughput screen was performed in order to identify chemotypes that are bound by AMPK. A novel thienopyridone compound (1) was identified and subsequently optimized. The structure-activity relationships that emerged from this effort are described.
Subject(s)
Energy Metabolism/drug effects , Enzyme Activation/drug effects , Enzyme Activators/pharmacology , Multienzyme Complexes/metabolism , Protein Serine-Threonine Kinases/metabolism , Pyridines/pharmacology , AMP-Activated Protein Kinases , Biological Assay , Energy Metabolism/physiology , Enzyme Activation/physiology , Enzyme Activators/chemistry , Pyridines/chemistry , Structure-Activity RelationshipABSTRACT
A series of potent 2-carboxychromone-based melanin-concentrating hormone receptor 1 (MCHr1) antagonists were synthesized and evaluated for hERG (human Ether-a-go-go Related Gene) channel affinity and functional blockade. Basic dialkylamine-terminated analogs were found to weakly bind the hERG channel and provided marked improvement in a functional patch-clamp assay versus previously reported antagonists of the series.
Subject(s)
Amides/pharmacology , Chromones/pharmacology , Ether-A-Go-Go Potassium Channels/metabolism , Receptors, Pituitary Hormone/antagonists & inhibitors , Animals , Ether-A-Go-Go Potassium Channels/drug effects , Humans , Inhibitory Concentration 50 , Mice , Obesity/drug therapy , Patch-Clamp Techniques , PharmacokineticsABSTRACT
The optimization of potent MCHr1 antagonist 1 with respect to improving its in vitro profile by replacement of the 3,4-methylenedioxy phenyl (piperonyl) moiety led to the discovery of 19, a compound that showed excellent MCHr1 binding and functional potencies in addition to possessing superior hERG separation, CYP3A4 profile, and receptor cross-reactivity profiles.
Subject(s)
Piperidines/chemical synthesis , Piperidines/pharmacology , Receptors, Somatostatin/antagonists & inhibitors , Alkylation , Animals , Chemical Phenomena , Chemistry, Physical , Chromones , Cross Reactions , Cytochrome P-450 CYP3A , Cytochrome P-450 Enzyme Inhibitors , Dose-Response Relationship, Drug , ERG1 Potassium Channel , Enzyme Inhibitors/chemical synthesis , Enzyme Inhibitors/pharmacology , Ether-A-Go-Go Potassium Channels/pharmacology , Heart Rate/drug effects , Heterocyclic Compounds/chemical synthesis , Heterocyclic Compounds/pharmacology , Humans , Mice , Structure-Activity RelationshipABSTRACT
The incorporation of constrained tertiary amines into an existing class of N-benzyl-4-aminopiperidinyl chromone-based MCHr1 antagonists led to the identification of a series of chiral racemic compounds that displayed good to excellent functional potency, binding affinity, and selectivity over the hERG channel. Further separation of two distinct chiral racemic compounds into their corresponding pairs of enantiomers revealed a considerable selectivity for MCHr1 for one configuration, in addition to a striking difference in oral exposure between one pair of enantiomers in diet-induced obese mice. Oral administration of the most potent compound in this class in the same animal model led to significant reduction of fat mass in a semi-chronic model for weight loss.