RESUMEN
The NOD-like receptor pyrin domain-containing protein 3 (NLRP3) inflammasome is a central regulator of innate immunity, essential for processing and release of interleukin-1ß and pyroptotic cell death. As endogenous NLRP3 activating triggers are hallmarks of many human chronic inflammatory diseases, inhibition of NLRP3 has emerged as a therapeutic target. Here we identify NDT-19795 as a novel carboxylic acid-containing NLRP3 activation inhibitor in both human and mouse monocytes and macrophages. Remarkably, conversion of the carboxylate to an isopropyl-ester (NT-0796) greatly enhances NLRP3 inhibitory potency in human monocytes. This increase is attributed to the ester-containing pharmacophore being more cell-penetrant than the acid species and, once internalized, the ester being metabolized to NDT-19795 by carboxylesterase-1 (CES-1). Mouse macrophages do not express CES-1, and NT-0796 is ineffective in these cells. Mice also contain plasma esterase (Ces1c) activity which is absent in humans. To create a more human-like model, we generated a mouse line in which the genome was modified, removing Ces1c and replacing this segment of DNA with the human CES-1 gene driven by a mononuclear phagocyte-specific promoter. We show human CES-1 presence in monocytes/macrophages increases the ability of NT-0796 to inhibit NLRP3 activation both in vitro and in vivo. As NLRP3 is widely expressed by monocytes/macrophages, the co-existence of CES-1 in these same cells affords a unique opportunity to direct ester-containing NLRP3 inhibitors precisely to target cells of interest. Profiling NT-0796 in mice humanized with respect to CES-1 biology enables critical modeling of the pharmacokinetics and pharmacodynamics of this novel therapeutic candidate. SIGNIFICANCE STATEMENT: Inhibition of NLRP3 represents a desirable therapeutic strategy for the treatment of multiple human disorders. In this study pharmacological properties of a structurally-novel, ester-containing NLRP3 inhibitor NT-0796 are characterized. To study pharmacodynamics of NT-0796 in vivo, a mouse line was engineered possessing more human-like traits with respect to carboxylesterase biology. In the context of these hCES-1 mice, NT-0796 serves as a more effective inhibitor of NLRP3 activation than the corresponding acid, highlighting the full translational potential of the ester strategy.
Asunto(s)
Proteína con Dominio Pirina 3 de la Familia NLR , Proteínas NLR , Humanos , Animales , Ratones , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Dominio Pirina , Inflamasomas/metabolismo , Caspasa 1/metabolismo , Ésteres , Hidrolasas de Éster Carboxílico/metabolismo , Interleucina-1beta/metabolismoRESUMEN
The NLRP3 inflammasome is a component of the innate immune system involved in the production of proinflammatory cytokines. Neurodegenerative disorders, including Alzheimer's disease, Parkinson's disease, multiple sclerosis, and amyotrophic lateral sclerosis, have been shown to have a component driven by NLRP3 inflammasome activation. Diseases such as these with large unmet medical needs have resulted in an interest in inhibiting the NLRP3 inflammasome as a potential pharmacological treatment, but to date, no marketed drugs specifically targeting NLRP3 have been approved. Furthermore, the requirement for CNS-penetrant molecules adds additional complexity to the search for NLRP3 inflammasome inhibitors suitable for clinical investigation of neuroinflammatory disorders. We designed a series of ester-substituted carbamate compounds as selective NLRP3 inflammasome inhibitors, leading to NT-0796, an isopropyl ester that undergoes intracellular conversion to NDT-19795, the carboxylic acid active species. NT-0796 was shown to be a potent and selective NLRP3 inflammasome inhibitor with demonstrated in vivo brain penetration.
Asunto(s)
Inflamasomas , Proteína con Dominio Pirina 3 de la Familia NLR , Humanos , Inflamasomas/metabolismo , Proteína con Dominio Pirina 3 de la Familia NLR/fisiología , Enfermedades Neuroinflamatorias , Encéfalo/metabolismo , ÉsteresRESUMEN
The NLRP3 inflammasome is a multiprotein complex that facilitates activation and release of the proinflammatory cytokines interleukin-1ß (IL-1ß) and IL-18 in response to infection or endogenous stimuli. It can be inappropriately activated by a range of danger signals resulting in chronic, low-grade inflammation underlying a multitude of diseases, such as Alzheimer's disease, Parkinson's disease, osteoarthritis, and gout. The discovery of potent and specific NLRP3 inhibitors could reduce the burden of several common morbidities. In this study, we identified a weakly potent triazolopyrimidone hit (1) following an in silico modeling exercise. This was optimized to furnish potent and selective small molecule NLRP3 inflammasome inhibitors. Compounds such as NDT-30805 could be useful tool molecules for a scaffold-hopping or pharmacophore generation project or used as leads toward the development of clinical candidates.
RESUMEN
Histamine H3 receptor (H3R) inverse agonists that have been in clinical trials for the treatment of excessive sleep disorders, have been plagued with insomnia as a mechanism-based side effect. We focused on the identification of compounds that achieve high receptor occupancy within a short time, followed by rapid disengagement from the receptor, a target profile that could provide therapeutic benefits without the undesired side effect of insomnia. This article describes the optimization work that led to the discovery of 1-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)piperidin-4-yl 4-cyclobutylpiperazine-1-carboxylate (18 b, LML134).
Asunto(s)
Agonistas de los Receptores Histamínicos/uso terapéutico , Piperazina/química , Piperazinas/química , Receptores Histamínicos H3/metabolismo , Trastornos del Sueño-Vigilia/tratamiento farmacológico , Animales , Evaluación Preclínica de Medicamentos , Agonismo Inverso de Drogas , Semivida , Agonistas de los Receptores Histamínicos/química , Agonistas de los Receptores Histamínicos/farmacocinética , Humanos , Masculino , Microsomas Hepáticos/metabolismo , Piperazina/farmacocinética , Piperazina/uso terapéutico , Piperazinas/farmacocinética , Piperazinas/uso terapéutico , Ratas , Ratas Sprague-Dawley , Receptores Histamínicos H3/química , Relación Estructura-ActividadRESUMEN
Ergolines were recently identified as a novel class of H3 receptor (H3R) inverse agonists. Although their optimization led to drug candidates with encouraging properties for the treatment of narcolepsy, brain penetration remained low. To overcome this issue, ergoline 1 ((6aR,9R,10aR)-4-(2-(dimethylamino)ethyl)-N-phenyl-9-(pyrrolidine-1-carbonyl)-6,6a,8,9,10,10a-hexahydroindolo[4,3-fg]quinoline-7(4H)-carboxamide)) was transformed into a series of indole derivatives with high H3R affinity. These new molecules were profiled by simultaneous determination of their brain receptor occupancy (RO) levels and pharmacodynamic (PD) effects in mice. These efforts culminated in the discovery of 15 m ((R)-1-isopropyl-5-(1-(2-(2-methylpyrrolidin-1-yl)ethyl)-1H-indol-4-yl)pyridin-2(1H)-one), which has an ideal profile showing a strong correlation of PD effects with RO, and no measurable safety liabilities. Its desirably short duration of action was confirmed by electroencephalography (EEG) measurements in rats.
Asunto(s)
Ergolinas/química , Antagonistas de los Receptores Histamínicos/química , Indoles/química , Piridonas/química , Receptores Histamínicos H3/química , Animales , Encéfalo/metabolismo , Células CHO , Cricetinae , Cricetulus , Electroencefalografía , Ergolinas/farmacocinética , Ergolinas/uso terapéutico , Semivida , Antagonistas de los Receptores Histamínicos/farmacocinética , Antagonistas de los Receptores Histamínicos/uso terapéutico , Humanos , Indoles/farmacocinética , Indoles/uso terapéutico , Masculino , Ratones , Narcolepsia/tratamiento farmacológico , Narcolepsia/metabolismo , Narcolepsia/patología , Unión Proteica , Piridonas/farmacocinética , Piridonas/uso terapéutico , Ratas , Ratas Sprague-Dawley , Receptores Histamínicos H3/metabolismo , Relación Estructura-ActividadRESUMEN
We describe the synthesis and characterization of 3-alkoxy-pyrrolo[1,2-b]pyrazolines as novel selective androgen receptor (AR) modulators that possess excellent physicochemical properties for transdermal administration. Compound 26 bound to human AR with an IC50 of 0.7 nM with great selectivity over other nuclear hormone receptors and potently activated AR in a C2C12 muscle cell reporter gene assay with an EC50 of 0.5 nM. It showed high aqueous solubility of 1.3 g/L at pH 7.4, and an in silico model as well as a customized parallel artificial membrane permeability assay indicated good skin permeation. Indeed, when measuring skin permeation through excised human skin, an excellent flux of 2 µg/(cm(2)·h) was determined without any permeation enhancers. In a 2 week Hershberger model using castrated rats, the compound showed dose-dependent effects fully restoring skeletal muscle weight at 0.3 mg/kg/day after subcutaneous administration with high selectivity over prostate stimulation.
Asunto(s)
Antagonistas de Receptores Androgénicos/química , Andrógenos/química , Compuestos de Azabiciclo/química , Pirazoles/química , Receptores Androgénicos/química , Administración Cutánea , Antagonistas de Receptores Androgénicos/metabolismo , Antagonistas de Receptores Androgénicos/farmacocinética , Andrógenos/metabolismo , Animales , Área Bajo la Curva , Compuestos de Azabiciclo/metabolismo , Compuestos de Azabiciclo/farmacocinética , Sitios de Unión , Unión Competitiva , Línea Celular , Fenómenos Químicos , Cristalografía por Rayos X , Humanos , Masculino , Tasa de Depuración Metabólica , Ratones , Modelos Químicos , Modelos Moleculares , Estructura Molecular , Estructura Terciaria de Proteína , Pirazoles/metabolismo , Pirazoles/farmacocinética , Ratas Wistar , Receptores Androgénicos/metabolismo , Piel/metabolismoRESUMEN
Ergoline derivative (6aR,9R)-4-(2-(dimethylamino)ethyl)-N-phenyl-9-(pyrrolidine-1-carbonyl)-6,6a,8,9-tetrahydroindolo[4,3-fg]quinoline-7(4H)-carboxamide (1), a CXCR3 antagonist, also inhibits human histamine H3 receptors (H3R) and represents a structurally novel H3R inverse agonist chemotype. It displays favorable pharmacokinetic and in vitro safety profiles, and served as a lead compound in a program to explore ergoline derivatives as potential drug candidates for the treatment of narcolepsy. A key objective of this work was to enhance the safety and efficacy profiles of 1, while minimizing its duration of action to mitigate the episodes of insomnia documented with previously reported clinical candidates during the night following administration. Modifications to the ergoline core at positionsâ 1, 6 and 8 were systematically investigated, and derivative 23 (1-((4aR,8R,9aR)-8-(hydroxymethyl)-1-(2-((R)-2-methylpyrrolidin-1-yl)ethyl)-4,4a,7,8,9,9a-hexahydroindolo[1,14-fg]quinolin-6(1H)-yl)ethanone) was identified as a promising lead compound. Derivative 23 has a desirable pharmacokinetic profile and demonstrated efficacy by enhancing brain concentrations of tele-methylhistamine, a major histamine metabolite. This validates the potential of the ergoline scaffold to serve as a template for the development of H3R inverse agonists.
Asunto(s)
Ergolinas/química , Agonistas de los Receptores Histamínicos/química , Receptores Histamínicos H3/química , Animales , Células CACO-2 , Línea Celular , Perros , Agonismo Inverso de Drogas , Ergolinas/farmacocinética , Ergolinas/uso terapéutico , Semivida , Agonistas de los Receptores Histamínicos/farmacocinética , Agonistas de los Receptores Histamínicos/uso terapéutico , Humanos , Células de Riñón Canino Madin Darby , Masculino , Ratones , Microsomas Hepáticos/metabolismo , Narcolepsia/tratamiento farmacológico , Unión Proteica , Ratas , Ratas Sprague-Dawley , Receptores Histamínicos H3/metabolismo , Relación Estructura-ActividadRESUMEN
A new class of indazole-derived bradykinin B(1) antagonists and their structure-activity relationships (SAR) is reported. A number of compounds were found to have low-nanomolar affinity for the human B(1) receptor and possess acceptable P-gp and pharmacokinetics properties.
Asunto(s)
Antagonistas del Receptor de Bradiquinina B1 , Indazoles/farmacología , Miembro 1 de la Subfamilia B de Casetes de Unión a ATP , Humanos , Indazoles/farmacocinética , Unión Proteica , Relación Estructura-ActividadRESUMEN
The discovery and synthesis of 4,4-disubstituted quinazolinones as T-type calcium channel antagonists is reported. Based on lead compounds 2 and 3, a focused SAR campaign driven by the optimization of potency, metabolic stability, and pharmacokinetic profile identified 45 as a potent T-type Ca(2+) channel antagonist with minimized PXR activation. In vivo, 45 suppressed seizure frequency in a rat model of absence epilepsy and showed significant alterations of sleep architecture after oral dosing to rats as measured by EEG.
Asunto(s)
Bloqueadores de los Canales de Calcio/farmacología , Canales de Calcio Tipo T/efectos de los fármacos , Quinazolinonas/química , Quinazolinonas/farmacología , Animales , Disponibilidad Biológica , Bloqueadores de los Canales de Calcio/química , Bloqueadores de los Canales de Calcio/farmacocinética , Cromatografía Líquida de Alta Presión , Descubrimiento de Drogas , Haplorrinos , Humanos , Quinazolinonas/farmacocinética , Ratas , Relación Estructura-ActividadRESUMEN
A novel series of quinazolinone T-type calcium channel antagonists have been prepared and evaluated using in vitro and in vivo assays. Optimization of the screening hit 3 by modifications of the 3- and 4-positions of the quinazolinone ring afforded potent and selective antagonists that displayed in vivo central nervous system efficacy in epilepsy and tremor models, as well as significant effects on rat active wake as measured by electrocorticogram.
RESUMEN
A series of amiloride derivatives modified at the 5-position of the pyrazine ring were evaluated as inhibitors of acid-sensing ion channel-3 (ASIC3), a novel target for the treatment of chronic pain.
Asunto(s)
Amilorida/análogos & derivados , Química Farmacéutica/métodos , Proteínas del Tejido Nervioso/química , Dolor/tratamiento farmacológico , Canales de Sodio/química , Canales Iónicos Sensibles al Ácido , Acidosis , Amilorida/química , Aminas/química , Animales , Diseño de Fármacos , Electrofisiología , Concentración 50 Inhibidora , Masculino , Modelos Químicos , Pirazinas/química , Ratas , Ratas Sprague-DawleyRESUMEN
Bradykinin B(1) receptor antagonists embody a potentially novel approach for the treatment of chronic pain and inflammation. The identification of non-peptide B(1) antagonists has been a notable advance in the kinin field that will allow evaluation of their therapeutic potential in the clinical realm. The current review is a high level summary of our contributions to the area that culminated in the discovery of a clinical candidate.
Asunto(s)
Analgésicos/química , Analgésicos/farmacología , Antagonistas del Receptor de Bradiquinina B1 , Analgésicos/uso terapéutico , Animales , Química Farmacéutica , Humanos , Inflamación/tratamiento farmacológico , Dolor/tratamiento farmacológico , Receptor de Bradiquinina B1/metabolismo , Relación Estructura-ActividadRESUMEN
The discovery of a novel series of potent and selective T-type calcium channel antagonists is reported. Initial optimization of high-throughput screening leads afforded a 1,4-substituted piperidine amide 6 with good potency and limited selectivity over hERG and L-type channels and other off-target activities. Further SAR on reducing the basicity of the piperidine and introducing polarity led to the discovery of 3-axial fluoropiperidine 30 with a significantly improved selectivity profile. Compound 30 showed good oral bioavailability and brain penetration across species. In a rat genetic model of absence epilepsy, compound 30 demonstrated a robust reduction in the number and duration of seizures at 33 nM plasma concentration, with no cardiovascular effects at up to 5.6 microM. Compound 30 also showed good efficacy in rodent models of essential tremor and Parkinson's disease. Compound 30 thus demonstrates a wide margin between CNS and peripheral effects and is a useful tool for probing the effects of T-type calcium channel inhibition.
Asunto(s)
Bloqueadores de los Canales de Calcio/síntesis química , Bloqueadores de los Canales de Calcio/farmacología , Canales de Calcio Tipo T/metabolismo , Piperidinas/síntesis química , Piperidinas/farmacología , Animales , Bloqueadores de los Canales de Calcio/química , Sistema Cardiovascular/efectos de los fármacos , Evaluación Preclínica de Medicamentos , Humanos , Estructura Molecular , Piperidinas/química , Ratas , Relación Estructura-ActividadRESUMEN
A series of carbo- and heterocyclic alpha-hydroxy amide-derived bradykinin B1 antagonists was prepared and evaluated. A 4,4-difluorocyclohexyl alpha-hydroxy amide was incorporated along with a 2-methyl tetrazole in lieu of an oxadiazole to afford a suitable compound with good pharmacokinetic properties, CNS penetration, and clearance by multiple metabolic pathways.
Asunto(s)
Amidas/síntesis química , Amidas/farmacología , Antagonistas del Receptor de Bradiquinina B1 , Tetrazoles/síntesis química , Tetrazoles/farmacología , Amidas/química , Amidas/farmacocinética , Animales , Sistema Nervioso Central/efectos de los fármacos , Técnicas Químicas Combinatorias , Diseño de Fármacos , Humanos , Estructura Molecular , Ratas , Relación Estructura-Actividad , Tetrazoles/química , Tetrazoles/farmacocinéticaRESUMEN
The novel T-type antagonist ( S)- 5 has been prepared and evaluated in in vitro and in vivo assays for T-type calcium ion channel activity. Structural modification of the piperidine leads 1 and 2 afforded the fluorinated piperidine ( S)- 5, a potent and selective antagonist that displayed in vivo CNS efficacy without adverse cardiovascular effects.
Asunto(s)
Bloqueadores de los Canales de Calcio/síntesis química , Bloqueadores de los Canales de Calcio/farmacología , Canales de Calcio Tipo T/metabolismo , Diseño de Fármacos , Piperidinas/síntesis química , Piperidinas/farmacología , Piranos/síntesis química , Piranos/farmacología , Animales , Presión Sanguínea/efectos de los fármacos , Bloqueadores de los Canales de Calcio/química , Perros , Relación Dosis-Respuesta a Droga , Haplorrinos , Frecuencia Cardíaca/efectos de los fármacos , Modelos Animales , Estructura Molecular , Piperidinas/química , Piranos/química , Ratas , Relación Estructura-ActividadRESUMEN
Selective bradykinin (BK) B 1 receptor antagonists could be novel therapeutic agents for the treatment of pain and inflammation. Elucidation of the structure activity relationships of the structurally novel HTS lead compound 1 provided potent hBK B 1 receptor antagonists with excellent receptor occupancy in the CNS of hBK B 1 transgenic rats.
Asunto(s)
Aminas/química , Benzofenonas/química , Benzofenonas/farmacología , Antagonistas del Receptor de Bradiquinina B1 , Animales , Benzofenonas/síntesis química , Línea Celular , Perros , Humanos , Estructura Molecular , Ratas , Receptor de Bradiquinina B1/metabolismo , Relación Estructura-ActividadRESUMEN
After oral treatment (once daily) for 4 weeks with the potent bradykinin B(1) receptor antagonist methyl 3-chloro-3'-fluoro-4'-{(1R)-1-[({1-[(trifluoroacetyl)amino]cyclopropyl}carbonyl)-amino]ethyl}-1,1'-biphenyl-2-carboxylate (MK-0686), rhesus monkeys (Macaca mulatta) exhibited significantly reduced systemic exposure of the compound in a dose-dependent manner, suggesting an occurrence of autoinduction of MK-0686 metabolism. This possibility is supported by two observations. 1) MK-0686 was primarily eliminated via biotransformation in rhesus monkeys, with oxidation on the chlorophenyl ring as one of the major metabolic pathways. This reaction led to appreciable formation of a dihydrodiol (M11) and a hydroxyl (M13) product in rhesus liver microsomes supplemented with NADPH. 2) The formation rate of these two metabolites determined in liver microsomes from MK-0686-treated groups was > or = 2-fold greater than the value for a control group. Studies with recombinant rhesus P450s and monoclonal antibodies against human P450 enzymes suggested that CYP2C75 played an important role in the formation of M11 and M13. The induction of this enzyme by MK-0686 was further confirmed by a concentration-dependent increase of its mRNA in rhesus hepatocytes, and, more convincingly, the enhanced CYP2C proteins and catalytic activities toward CYP2C75 probe substrates in liver microsomes from MK-0686-treated animals. Furthermore, a good correlation was observed between the rates of M11 and M13 formation and hydroxylase activities toward probe substrates determined in a panel of liver microsomal preparations from control and MK-0686-treated animals. Therefore, MK-0686, both a substrate and inducer for CYP2C75, caused autoinduction of its own metabolism in rhesus monkeys by increasing the expression of this enzyme.
Asunto(s)
Acetamidas/farmacocinética , Benzoatos/farmacocinética , Antagonistas del Receptor de Bradiquinina B1 , Sistema Enzimático del Citocromo P-450/metabolismo , Acetamidas/sangre , Acetamidas/orina , Animales , Benzoatos/sangre , Benzoatos/orina , Bilis/metabolismo , Línea Celular Tumoral , Células Cultivadas , Sistema Enzimático del Citocromo P-450/genética , Femenino , Hepatocitos/metabolismo , Humanos , Macaca mulatta , Masculino , Microsomas Hepáticos/metabolismo , Receptor X de Pregnano , Receptor de Bradiquinina B1/metabolismo , Receptores de Esteroides/metabolismo , Proteínas Recombinantes/metabolismoRESUMEN
The design and synthesis of a novel class of human bradykinin B1 antagonists featuring difluoroethyl ether and isoxazole carboxamide moieties are disclosed. Compound 7g displayed excellent pharmacokinetic properties, efficient ex vivo receptor occupancy, and low potential for P450 induction via PXR activation.
Asunto(s)
Antagonistas del Receptor de Bradiquinina B1 , Isoxazoles/farmacología , Receptores de Esteroides/efectos de los fármacos , Administración Oral , Animales , Disponibilidad Biológica , Perros , Humanos , Isoxazoles/farmacocinética , Macaca mulatta , Receptor X de Pregnano , Ratas , Ratas Sprague-DawleyRESUMEN
Antagonism of the bradykinin B(1) receptor represents a potential treatment for chronic pain and inflammation. Novel antagonists incorporating alpha-hydroxy amides were designed that display low-nanomolar affinity for the human bradykinin B(1) receptor and good bioavailability in the rat and dog. In addition, these functionally active compounds show high passive permeability and low susceptibility to phosphoglycoprotein mediated efflux, predictive of good CNS exposure.
Asunto(s)
Amidas/farmacología , Antagonistas del Receptor de Bradiquinina B1 , Amidas/química , Amidas/farmacocinética , Animales , Disponibilidad Biológica , Barrera Hematoencefálica , Inhibidores Enzimáticos del Citocromo P-450 , Perros , Semivida , Humanos , Ratas , Ratas Sprague-Dawley , Relación Estructura-ActividadRESUMEN
SAR study of the biphenyl region of cyclopropanecarboxamide derived bradykinin B(1) antagonists was examined. Incorporation of a pyridine in place of the proximal phenyl ring and chlorination of the distal phenyl ring proved to be well tolerated and provided compounds with improved pharmacokinetic profiles, CNS penetration, and enhanced receptor occupancy.