RESUMEN

Multiple sclerosis (MS) is a chronic and debilitating neurological disease that results in inflammatory demyelination. While endogenous remyelination helps to recover function, this restorative process tends to become less efficient over time. Currently, intense efforts aimed at the mechanisms that promote remyelination are being considered promising therapeutic approaches. The M1 muscarinic acetylcholine receptor (M1R) was previously identified as a negative regulator of oligodendrocyte differentiation and myelination. Here, we validate M1R as a target for remyelination by characterizing expression in human and rodent oligodendroglial cells (including those in human MS tissue) using a highly selective M1R probe. As a breakthrough to conventional methodology, we conjugated a fluorophore to a highly M1R selective peptide (MT7) which targets the M1R in the subnanomolar range. This allows for exceptional detection of M1R protein expression in the human CNS. More importantly, we introduce PIPE-307, a brain-penetrant, small-molecule antagonist with favorable drug-like properties that selectively targets M1R. We evaluate PIPE-307 in a series of in vitro and in vivo studies to characterize potency and selectivity for M1R over M2-5R and confirm the sufficiency of blocking this receptor to promote differentiation and remyelination. Further, PIPE-307 displays significant efficacy in the mouse experimental autoimmune encephalomyelitis model of MS as evaluated by quantifying disability, histology, electron microscopy, and visual evoked potentials. Together, these findings support targeting M1R for remyelination and support further development of PIPE-307 for clinical studies.

Asunto(s)

Esclerosis Múltiple , Oligodendroglía , Receptor Muscarínico M1 , Remielinización , Animales , Humanos , Ratones , Ratas , Encéfalo/metabolismo , Encéfalo/efectos de los fármacos , Encéfalo/patología , Encefalomielitis Autoinmune Experimental/tratamiento farmacológico , Encefalomielitis Autoinmune Experimental/metabolismo , Ratones Endogámicos C57BL , Esclerosis Múltiple/tratamiento farmacológico , Esclerosis Múltiple/metabolismo , Antagonistas Muscarínicos/farmacología , Vaina de Mielina/metabolismo , Oligodendroglía/metabolismo , Oligodendroglía/efectos de los fármacos , Receptor Muscarínico M1/metabolismo , Receptor Muscarínico M1/antagonistas & inhibidores , Remielinización/efectos de los fármacos

RESUMEN

Multiple sclerosis (MS) is a chronic neurological disease characterized by inflammatory demyelination that disrupts neuronal transmission resulting in neurodegeneration progressive disability. While current treatments focus on immunosuppression to limit inflammation and further myelin loss, no approved therapies effectively promote remyelination to mitigate the progressive disability associated with chronic demyelination. Lysophosphatidic acid (LPA) is a pro-inflammatory lipid that is upregulated in MS patient plasma and cerebrospinal fluid (CSF). LPA activates the LPA1 receptor, resulting in elevated CNS cytokine and chemokine levels, infiltration of immune cells, and microglial/astrocyte activation. This results in a neuroinflammatory response leading to demyelination and suppressed remyelination. A medicinal chemistry effort identified PIPE-791, an oral, brain-penetrant, LPA1 antagonist. PIPE-791 was characterized in vitro and in vivo and was found to be a potent, selective LPA1 antagonist with slow receptor off-rate kinetics. In vitro, PIPE-791 induced OPC differentiation and promoted remyelination following a demyelinating insult. PIPE-791 further mitigated the macrophage-mediated inhibition of OPC differentiation and inhibited microglial and fibroblast activation. In vivo, the compound readily crossed the blood-brain barrier and blocked LPA1 in the CNS after oral dosing. Direct dosing of PIPE-791 in vivo increased oligodendrocyte number, and in the mouse experimental autoimmune encephalomyelitis (EAE) model of MS, we observed that PIPE-791 promoted myelination, reduced neuroinflammation, and restored visual evoked potential latencies (VEP). These findings support targeting LPA1 for remyelination and encourage development of PIPE-791 for treating MS patients with advantages not seen with current immunosuppressive disease modifying therapies.

Asunto(s)

Esclerosis Múltiple , Receptores del Ácido Lisofosfatídico , Remielinización , Animales , Esclerosis Múltiple/tratamiento farmacológico , Esclerosis Múltiple/metabolismo , Receptores del Ácido Lisofosfatídico/antagonistas & inhibidores , Receptores del Ácido Lisofosfatídico/metabolismo , Remielinización/efectos de los fármacos , Humanos , Ratones , Enfermedades Neuroinflamatorias/tratamiento farmacológico , Enfermedades Neuroinflamatorias/metabolismo , Oligodendroglía/metabolismo , Oligodendroglía/efectos de los fármacos , Encéfalo/metabolismo , Encéfalo/efectos de los fármacos , Encéfalo/patología , Diferenciación Celular/efectos de los fármacos , Encefalomielitis Autoinmune Experimental/tratamiento farmacológico , Encefalomielitis Autoinmune Experimental/metabolismo , Ratones Endogámicos C57BL , Vaina de Mielina/metabolismo , Vaina de Mielina/efectos de los fármacos , Lisofosfolípidos/metabolismo , Barrera Hematoencefálica/metabolismo , Barrera Hematoencefálica/efectos de los fármacos

RESUMEN

Structure-activity relationship studies led to the discovery of PIPE-3297, a fully efficacious and selective kappa opioid receptor (KOR) agonist. PIPE-3297, a potent activator of G-protein signaling (GTPγS EC50 = 1.1 nM, 91% Emax), did not elicit a ß-arrestin-2 recruitment functional response (Emax < 10%). Receptor occupancy experiments performed with the novel KOR radiotracer [3H]-PIPE-3113 revealed that subcutaneous (s.c.) administration of PIPE-3297 at 30 mg/kg in mice achieved 90% occupancy of the KOR in the CNS 1 h post dose. A single subcutaneous dose of PIPE-3297 in healthy mice produced a statistically significant increase of mature oligodendrocytes (P < 0.0001) in the KOR-enriched striatum, an effect that was not observed in animals predosed with the selective KOR antagonist norbinaltorphimine. An equivalent dose given to mice in an open-field activity-monitoring system revealed a small KOR-independent decrease in total locomotor activity versus vehicle measured between 60 and 75 min post dose. Daily doses of PIPE-3297 at both 3 and 30 mg/kg s.c. reduced the disease score in the mouse experimental autoimmune encephalomyelitis (EAE) model. Visually evoked potential (VEP) N1 latencies were also significantly improved versus vehicle in both dose groups, and latencies matched those of untreated animals. Taken together, these findings highlight the potential therapeutic value of functionally selective G-protein KOR agonists in demyelinating disease, which may avoid the sedating side effects typically associated with classical nonbiased KOR agonists.

Asunto(s)

Receptores Opioides kappa , Transducción de Señal , Ratones , Animales , Arrestina beta 2/farmacología , Receptores Opioides kappa/agonistas , Proteínas de Unión al GTP/metabolismo , Antagonistas de Narcóticos/farmacología , Analgésicos Opioides/farmacología

RESUMEN

The discovery of PIPE-359, a brain-penetrant and selective antagonist of the muscarinic acetylcholine receptor subtype 1 is described. Starting from a literature-reported M1 antagonist, linker replacement and structure-activity relationship investigations of the eastern 1-(pyridinyl)piperazine led to the identification of a novel, potent, and selective antagonist with good MDCKII-MDR1 permeability. Continued semi-iterative positional scanning facilitated improvements in the metabolic and hERG profiles, which ultimately delivered PIPE-359. This advanced drug candidate exhibited robust efficacy in mouse myelin oligodendrocyte glycoprotein (MOG)-induced experimental autoimmune encephalitis (EAE), a preclinical model for multiple sclerosis.

RESUMEN

We previously published on the design and synthesis of novel, potent and selective PPARα antagonists suitable for either i.p. or oral in vivo administration for the potential treatment of cancer. Described herein is SAR for a subsequent program, where we set out to identify selective and potent PPARα/δ dual antagonist molecules. Emerging literature indicates that both PPARα and PPARδ antagonism may be helpful in curbing the proliferation of certain types of cancer. This dual antagonism could also be used to study PPARs in other settings. After testing for selective and dual potency, off-target counter screening, metabolic stability, oral bioavailability and associated toxicity, compound 11, the first reported PPARα/δ dual antagonist was chosen for more advanced preclinical evaluation.

Asunto(s)

Antineoplásicos/farmacología , Descubrimiento de Drogas , Neoplasias Ováricas/tratamiento farmacológico , PPAR alfa/antagonistas & inhibidores , PPAR delta/antagonistas & inhibidores , Sulfonamidas/farmacología , Animales , Antineoplásicos/síntesis química , Antineoplásicos/química , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Perros , Relación Dosis-Respuesta a Droga , Ensayos de Selección de Medicamentos Antitumorales , Femenino , Humanos , Ratones , Estructura Molecular , Neoplasias Experimentales/tratamiento farmacológico , Neoplasias Experimentales/metabolismo , Neoplasias Experimentales/patología , Neoplasias Ováricas/metabolismo , Neoplasias Ováricas/patología , PPAR alfa/metabolismo , PPAR delta/metabolismo , Ratas , Relación Estructura-Actividad , Sulfonamidas/síntesis química , Sulfonamidas/química

RESUMEN

Peroxisome-proliferator activated receptors (PPAR) are members of the nuclear hormone receptor superfamily which regulate gene transcription. PPARα is a key regulator of lipid homeostasis and a negative regulator of inflammation. Under conditions of metabolic stress such as fasting or glucose deprivation, PPARα is upregulated in order to control gene expression necessary for processing alternate fuel sources (e.g. fatty acid oxidation) and thereby promote maintenance of cell viability. Clinically, PPARα expression is upregulated in diseased tissues such as melanoma, chronic lymphocytic leukemia, ovarian and prostate cancer. This may allow for cellular proliferation and metastasis. Importantly, genetic knockouts of PPARα have been shown to be protected against tumor growth in a variety of syngeneic tumors models. We hypothesized that a potent and selective PPARα antagonist could represent a novel cancer therapy. Early in our discovery research, we identified NXT629 (Bravo et al., 2014). Herein we describe the pharmacology of NXT629 and demonstrate that it is a potent and selective PPARα antagonist. We identify NXT629 as a valuable tool for use in in vivo assessment of PPARα due to its good systemic exposure following intraperitoneal injection. We explore the in vivo pharmacology of NXT629 and demonstrate that it is efficacious in pharmacodynamic models that are driven by PPARα. Finally, we probe the efficacy of NXT629 in disease models where PPARα knockouts have shown to be protected. We believe that PPARα antagonists will be beneficial in diseases such as ovarian cancer and melanoma where PPARα and fatty acid oxidation may be involved.

Asunto(s)

Aminopiridinas/farmacología , PPAR alfa/antagonistas & inhibidores , Sulfonamidas/farmacología , Aminopiridinas/farmacocinética , Animales , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Factores de Crecimiento de Fibroblastos/sangre , Humanos , Ratones , Metástasis de la Neoplasia , Neovascularización Fisiológica/efectos de los fármacos , Ratas , Sulfonamidas/farmacocinética

RESUMEN

The discovery and SAR of a novel series of potent and selective PPARα antagonists are herein described. Exploration of replacements for the labile acyl sulfonamide linker led to a biaryl sulfonamide series of which compound 33 proved to be suitable for further profiling in vivo. Compound 33 demonstrated excellent potency, selectivity against other nuclear hormone receptors, and good pharmacokinetics in mouse.

Asunto(s)

PPAR alfa/antagonistas & inhibidores , Sulfonamidas/química , Sulfonamidas/farmacología , Animales , Butiratos/química , Butiratos/farmacología , Humanos , Ratones , Estructura Molecular , Oxazoles/química , Oxazoles/farmacología , Compuestos de Fenilurea/química , Compuestos de Fenilurea/farmacología , Propionatos/química , Propionatos/farmacología , Relación Estructura-Actividad , Triazoles/química , Triazoles/farmacología , Tirosina/análogos & derivados , Tirosina/química , Tirosina/farmacología

RESUMEN

The potent 5-lipoxygenase-activating protein (FLAP) inhibitor 3-[3-tert-butylsulfanyl-1-[4-(6-ethoxypyridin-3-yl)benzyl]-5-(5-methylpyridin-2-ylmethoxy)-1H-indol-2-yl]-2,2-dimethylpropionic acid 11cc is described (AM803, now GSK2190915). Building upon AM103 (1) (Hutchinson et al. J. Med Chem.2009, 52, 5803-5815; Stock et al. Bioorg. Med. Chem. Lett. 2010, 20, 213-217; Stock et al. Bioorg. Med. Chem. Lett.2010, 20, 4598-4601), SAR studies centering around the pyridine moiety led to the discovery of compounds that exhibit significantly increased potency in a human whole blood assay measuring LTB(4) inhibition with longer drug preincubation times (15 min vs 5 h). Further studies identified 11cc with a potency of 2.9 nM in FLAP binding, an IC(50) of 76 nM for inhibition of LTB(4) in human blood (5 h incubation) and excellent preclinical toxicology and pharmacokinetics in rat and dog. 11cc also demonstrated an extended pharmacodynamic effect in a rodent bronchoalveolar lavage (BAL) model. This compound has successfully completed phase 1 clinical studies in healthy volunteers and is currently undergoing phase 2 trials in asthmatic patients.

Asunto(s)

Inhibidores de Proteína Activante de 5-Lipoxigenasa/síntesis química , Antiasmáticos/síntesis química , Indoles/síntesis química , Ácidos Pentanoicos/síntesis química , Inhibidores de Proteína Activante de 5-Lipoxigenasa/farmacocinética , Inhibidores de Proteína Activante de 5-Lipoxigenasa/farmacología , Administración Oral , Animales , Antiasmáticos/farmacocinética , Antiasmáticos/farmacología , Lavado Broncoalveolar , Inhibidores Enzimáticos del Citocromo P-450 , Perros , Femenino , Humanos , Técnicas In Vitro , Indoles/farmacocinética , Indoles/farmacología , Masculino , Ácidos Pentanoicos/farmacocinética , Ácidos Pentanoicos/farmacología , Ratas , Ratas Sprague-Dawley , Relación Estructura-Actividad

RESUMEN

Biphenylacetic acid (5) was identified through a library screen as an inhibitor of the prostaglandin D(2) receptor DP2 (CRTH2). Optimization for potency and pharmacokinetic properties led to a series of selective CRTH2 antagonists. Compounds demonstrated potency in a human DP2 binding assay and a human whole blood eosinophil shape change assay, as well as good oral bioavailability in rat and dog, and efficacy in a mouse model of allergic rhinitis following oral dosing.

Asunto(s)

Modelos Animales de Enfermedad , Descubrimiento de Drogas , Fenilacetatos/farmacología , Receptores Inmunológicos/antagonistas & inhibidores , Receptores de Prostaglandina/antagonistas & inhibidores , Rinitis Alérgica Perenne/tratamiento farmacológico , Animales , Disponibilidad Biológica , Perros , Ratones , Fenilacetatos/química , Fenilacetatos/farmacocinética , Fenilacetatos/uso terapéutico , Ratas

RESUMEN

The prostaglandin D(2) (PGD(2)) receptor type 2 (DP2) is a G protein-coupled receptor that has been shown to be involved in a variety of allergic diseases, including allergic rhinitis, asthma, and atopic dermatitis. In this study, we describe the preclinical pharmacological and pharmacokinetic properties of the small-molecule DP2 antagonist [2'-(3-benzyl-1-ethyl-ureidomethyl)-6-methoxy-4'-trifluoromethyl-biphenyl-3-yl]-acetic acid (AM211). We determine that AM211 has high affinity for human, mouse, rat, and guinea pig DP2 and it shows selectivity over other prostanoid receptors and enzymes. Antagonist activity of AM211 at the DP2 receptor was confirmed by inhibition of PGD(2)-stimulated guanosine 5'-O-[γ-thio]triphosphate binding to membranes expressing human DP2. A basophil activation assay and a whole-blood assay of eosinophil shape change were used to demonstrate the ability of AM211 to potently antagonize PGD(2)-stimulated functional responses in relevant human cells and in the context of a physiologically relevant environment. AM211 exhibits good oral bioavailability in rats and dogs and dose-dependently inhibits 13,14-dihydro-15-keto-PGD(2)-induced leukocytosis in a guinea pig pharmacodynamic assay. AM211 demonstrates efficacy in two animal models of allergic inflammation, including an ovalbumin-induced lung inflammation model in guinea pigs and an ovalbumin-induced mouse model of allergic rhinitis. AM211 represents a potent and selective antagonist of DP2 that may be used clinically to evaluate the role of DP2 in T helper 2-driven allergic inflammatory diseases.

Asunto(s)

Modelos Animales de Enfermedad , Compuestos de Metilurea/uso terapéutico , Fenilacetatos/uso terapéutico , Antagonistas de Prostaglandina/uso terapéutico , Receptores Inmunológicos/antagonistas & inhibidores , Receptores de Prostaglandina/antagonistas & inhibidores , Rinitis Alérgica Perenne/tratamiento farmacológico , Adulto , Animales , Perros , Femenino , Cobayas , Células HEK293 , Humanos , Hipersensibilidad/tratamiento farmacológico , Hipersensibilidad/inmunología , Hipersensibilidad/metabolismo , Masculino , Compuestos de Metilurea/química , Compuestos de Metilurea/farmacología , Ratones , Ratones Endogámicos BALB C , Fenilacetatos/química , Fenilacetatos/farmacología , Neumonía/tratamiento farmacológico , Neumonía/inmunología , Neumonía/metabolismo , Antagonistas de Prostaglandina/química , Antagonistas de Prostaglandina/farmacología , Unión Proteica/fisiología , Distribución Aleatoria , Ratas , Ratas Sprague-Dawley , Receptores Inmunológicos/inmunología , Receptores Inmunológicos/metabolismo , Receptores de Prostaglandina/inmunología , Receptores de Prostaglandina/metabolismo , Rinitis Alérgica Perenne/inmunología , Rinitis Alérgica Perenne/metabolismo

RESUMEN

Compound 21 (AM432) was identified as a potent and selective antagonist of the DP(2) receptor (CRTH2). Modification of a bi-aryl core identified a series of tri-aryl antagonists of which compound 21 proved a viable clinical candidate. AM432 shows excellent potency in a human whole blood eosinophil shape change assay with prolonged incubation, a comparatively long off-rate from the DP(2) receptor, excellent pharmacokinetics in dog and in vivo activity in two mouse models of inflammatory disease after oral dosing.

Asunto(s)

Fenilacetatos/química , Piridinas/química , Receptores Inmunológicos/antagonistas & inhibidores , Receptores de Prostaglandina/antagonistas & inhibidores , Administración Oral , Animales , Modelos Animales de Enfermedad , Perros , Eosinófilos/efectos de los fármacos , Eosinófilos/inmunología , Humanos , Inflamación/tratamiento farmacológico , Ratones , Fenilacetatos/farmacocinética , Fenilacetatos/uso terapéutico , Piridinas/farmacocinética , Piridinas/uso terapéutico , Receptores Inmunológicos/metabolismo , Receptores de Prostaglandina/metabolismo

RESUMEN

We evaluated the in vivo pharmacological properties of AM803 3-[3-tert-butylsulfanyl-1-[4-(6-ethoxy-pyridin-3-yl)-benzyl]-5-(5-methyl-pyridin-2-ylmethoxy)-1H-indol-2-yl]-2,2-dimethyl-propionic acid, a selective five-lipoxygenase-activating protein (FLAP) inhibitor, using rat and mouse models of acute inflammation. Oral administration of AM803 (1 mg/kg) resulted in sustained inhibition of ex vivo ionophore-challenged whole blood LTB4 biosynthesis with >90% inhibition for up to 12 h and an EC50 of approximately 7 nM. When rat lungs were challenged in vivo with calcium-ionophore, AM803 inhibited LTB4 and cysteinyl leukotriene (CysLT) production with ED50s of 0.12 mg/kg and 0.37 mg/kg, respectively. The inhibition measured 16 h following a single oral dose of 3 mg/kg was 86% and 41% for LTB4 and CysLTs, respectively. In an acute inflammation setting, AM803 dose-dependently reduced LTB4, CysLTs, plasma protein extravasation and neutrophil influx induced by peritoneal zymosan injection. Finally, AM803 increased survival time in mice exposed to a lethal intravenous injection of platelet activating factor (PAF). The magnitude of effect was similar to that of an inhibitor of five-lipoxygenase (5-LO) and LTA4 hydrolase but superior to a leukotriene CysLT1 receptor antagonist. In summary, AM803 is a novel, potent and selective FLAP inhibitor that has excellent pharmacodynamic properties in vivo and is effective in animal models of acute inflammation and in a model of lethal shock.

Asunto(s)

Proteínas Portadoras/antagonistas & inhibidores , Indoles/farmacología , Inflamación/metabolismo , Proteínas de la Membrana/antagonistas & inhibidores , Ácidos Pentanoicos/farmacología , Propionatos/farmacología , Proteínas Activadoras de la 5-Lipooxigenasa , Animales , Enfermedad Crónica , Cisteína/biosíntesis , Modelos Animales de Enfermedad , Femenino , Humanos , Indoles/farmacocinética , Indoles/uso terapéutico , Inflamación/tratamiento farmacológico , Leucotrieno B4/biosíntesis , Leucotrienos/biosíntesis , Pulmón/efectos de los fármacos , Pulmón/metabolismo , Masculino , Ratones , Ácidos Pentanoicos/farmacocinética , Ácidos Pentanoicos/uso terapéutico , Factor de Activación Plaquetaria/farmacología , Propionatos/farmacocinética , Propionatos/uso terapéutico , Ratas , Especificidad por Sustrato , Zimosan/farmacología

RESUMEN

AM643 (compound 6, 3-{3-tert-butylsulfanyl-1-[4-(5-methoxy-pyrimidin-2-yl)-benzyl]-5-(5-methyl-pyridin-2-ylmethoxy)-1H-indol-2-yl]-2,2-dimethyl-propionic acid) was identified as a potential candidate for formulation as a topical agent for the treatment of skin disorders involving leukotriene production. Dermal application of 6 using a prototypical vehicle in a murine ear arachidonic acid model showed significant reduction in the concentrations of leukotrienes in mouse skin with concomitant reduction in ear swelling.

Asunto(s)

Inhibidores Enzimáticos/química , Indoles/síntesis química , Propionatos/síntesis química , Proteínas Activadoras de la 5-Lipooxigenasa/metabolismo , Administración Tópica , Animales , Inhibidores Enzimáticos/síntesis química , Inhibidores Enzimáticos/uso terapéutico , Humanos , Indoles/química , Indoles/uso terapéutico , Leucotrienos/biosíntesis , Ratones , Propionatos/química , Propionatos/uso terapéutico , Ratas , Enfermedades de la Piel/inducido químicamente , Enfermedades de la Piel/tratamiento farmacológico

RESUMEN

Prostaglandin D(2) (PGD(2)) is one of a family of biologically active lipids derived from arachidonic acid via the action of COX-1 and COX-2. PGD(2) is released from mast cells and binds primarily to two G protein-coupled receptors, namely DP1 and DP2, the latter also known as chemoattractant receptor-homologous molecule expressed on Th2 cells. DP2 is predominantly expressed on eosinophils, Th2 cells, and basophils, but it is also expressed to a lesser extent on monocytes, mast cells, and epithelial cells. Interaction of PGD(2) and its active metabolites with DP2 results in cellular chemotaxis, degranulation, up-regulation of adhesion molecules, and cytokine production. Chronic obstructive pulmonary disease (COPD) is a chronic progressive inflammatory disease characterized by elevated lung neutrophils, macrophages, and CD8+ T lymphocytes and mucus hypersecretion. Cigarette smoke contributes to the etiology of COPD and was used here as a provoking agent in a murine model of COPD. In an acute model, {2'-[(cyclopropanecarbonyl-ethyl-amino)-methyl]-6-methoxy-4'-trifluoro-methyl-biphenyl-3-yl}-acetic acid, sodium salt (AM156) and (5-{2-[(benzoyloxycarbonyl-ethyl-amino)-methyl]-4-trifluoromethyl-phenyl}-pyridin-3-yl)-acetic acid, sodium salt) (AM206), potent DP2 receptor antagonists, dose-dependently inhibited influx of neutrophils and lymphocytes to smoke-exposed airways. In a subchronic model, AM156 and AM206 inhibited neutrophil and lymphocyte trafficking to the airways. Furthermore, AM156 and AM206 treatment inhibited mucus cell metaplasia and prevented the thickening of the airway epithelial layer induced by cigarette smoke. These data suggest that DP2 receptor antagonism may represent a novel therapy for COPD or other conditions characterized by neutrophil influx, mucus hypersecretion, and airway remodeling.

Asunto(s)

Pulmón/efectos de los fármacos , Moco/metabolismo , Enfermedad Pulmonar Obstructiva Crónica/prevención & control , Receptores Inmunológicos/antagonistas & inhibidores , Receptores de Prostaglandina/antagonistas & inhibidores , Mucosa Respiratoria/efectos de los fármacos , Fumar/efectos adversos , Animales , Bencilaminas/farmacocinética , Bencilaminas/farmacología , Línea Celular , Movimiento Celular , Femenino , Cobayas , Humanos , Técnicas In Vitro , Inflamación/inmunología , Inflamación/metabolismo , Inflamación/prevención & control , Pulmón/inmunología , Pulmón/patología , Linfocitos/efectos de los fármacos , Linfocitos/inmunología , Masculino , Metaplasia , Ratones , Ratones Endogámicos BALB C , Neutrófilos/efectos de los fármacos , Neutrófilos/inmunología , Niacina/análogos & derivados , Niacina/farmacocinética , Niacina/farmacología , Enfermedad Pulmonar Obstructiva Crónica/etiología , Enfermedad Pulmonar Obstructiva Crónica/inmunología , Enfermedad Pulmonar Obstructiva Crónica/patología , Mucosa Respiratoria/patología

RESUMEN

A series of potent 5-lipoxygenase-activating protein (FLAP) inhibitors are herein described. SAR studies focused on the discovery of novel alicyclic moieties appended to an indole core to optimize potency, physical properties and off-target activities. Subsequent SAR on the N-benzyl substituent of the indole led to the discovery of compound 39 (AM679) which showed potent inhibition of leukotrienes in human blood and in a rodent bronchoalvelolar lavage (BAL) challenge model.

Asunto(s)

Proteínas Portadoras/antagonistas & inhibidores , Indoles/química , Inhibidores de la Lipooxigenasa/química , Proteínas de la Membrana/antagonistas & inhibidores , Ácidos Pentanoicos/química , Proteínas Activadoras de la 5-Lipooxigenasa , Animales , Proteínas Portadoras/metabolismo , Humanos , Indoles/síntesis química , Indoles/farmacología , Leucotrienos/sangre , Leucotrienos/metabolismo , Inhibidores de la Lipooxigenasa/síntesis química , Inhibidores de la Lipooxigenasa/farmacología , Proteínas de la Membrana/metabolismo , Ratones , Modelos Animales , Ácidos Pentanoicos/síntesis química , Ácidos Pentanoicos/farmacología , Ratas , Relación Estructura-Actividad

RESUMEN

The potent and selective 5-lipoxygenase-activating protein leukotriene synthesis inhibitor 3-[3-tert-butylsulfanyl-1-[4-(6-methoxy-pyridin-3-yl)-benzyl]-5-(pyridin-2-ylmethoxy)-1H-indol-2-yl]-2,2-dimethyl-propionic acid (11j) is described. Lead optimization was designed to afford compounds with superior in vitro and in vivo inhibition of leukotriene synthesis in addition to having excellent pharmacokinetics and safety in rats and dogs. The key structural features of these new compounds are incorporation of heterocycles on the indole N-benzyl substituent and replacement of the quinoline group resulting in compounds with excellent in vitro and in vivo activities, superior pharmacokinetics, and improved physical properties. The methoxypyridine derivative 11j has an IC(50) of 4.2 nM in a 5-lipoxygenase-activating protein (FLAP) binding assay, an IC(50) of 349 nM in the human blood LTB(4) inhibition assay, and is efficacious in a murine ovalbumin model of allergen-induced asthma. Compound 11j was selected for clinical development and has successfully completed phase 1 trials in healthy volunteers.

Asunto(s)

Proteínas Portadoras/antagonistas & inhibidores , Indoles/farmacocinética , Leucotrieno B4/antagonistas & inhibidores , Proteínas de la Membrana/antagonistas & inhibidores , Propionatos/farmacocinética , Proteínas Activadoras de la 5-Lipooxigenasa , Animales , Asma/tratamiento farmacológico , Perros , Efectos Colaterales y Reacciones Adversas Relacionados con Medicamentos , Compuestos Heterocíclicos/química , Humanos , Concentración 50 Inhibidora , Leucotrieno B4/biosíntesis , Ratones , Unión Proteica , Ratas , Relación Estructura-Actividad

RESUMEN

Leukotrienes (LTs) are proinflammatory lipid mediators synthesized by the conversion of arachidonic acid (AA) to LTA(4) by the enzyme 5-lipoxygenase (5-LO) in the presence of 5-LO-activating protein (FLAP). 3-[3-tert-Butylsulfanyl-1-[4-(6-methoxy-pyridin-3-yl)-benzyl]-5-(pyridin-2-ylmethoxy)-1H-indol-2-yl]-2,2-dimethyl-propionic acid (AM103) is a novel selective FLAP inhibitor in development for the treatment of respiratory conditions such as asthma. In a rat ex vivo whole-blood calcium ionophore-induced LTB(4) assay, AM103 (administered orally at 1 mg/kg) displayed >50% inhibition for up to 6 h with a calculated EC(50) of approximately 60 nM. When rat lung was challenged in vivo with calcium ionophore, AM103 inhibited LTB(4) and cysteinyl leukotriene (CysLT) production with ED(50) values of 0.8 and 1 mg/kg, respectively. In this model, the EC(50) derived from plasma AM103 was approximately 330 nM for inhibition of both LTB(4) and CysLT. In an acute inflammation setting, AM103 displayed dose-dependent inhibition of LTB(4), CysLT, and plasma protein extravasation induced by peritoneal zymosan injection. In a model of chronic lung inflammation using ovalbumin-primed and challenged BALB/c mice, AM103 reduced the concentrations of eosinophil peroxidase, CysLTs, and interleukin-5 in the bronchoalveolar lavage fluid. Finally, AM103 increased survival time in mice exposed to a lethal intravenous injection of platelet-activating factor. In summary, AM103 is a novel, potent and selective FLAP inhibitor that has excellent pharmacodynamic properties in vivo and is effective in animal models of acute and chronic inflammation and in a model of lethal shock.

Asunto(s)

Antiinflamatorios no Esteroideos/farmacología , Proteínas Portadoras/antagonistas & inhibidores , Indoles/farmacología , Inflamación/tratamiento farmacológico , Proteínas de la Membrana/antagonistas & inhibidores , Propionatos/farmacología , Proteínas Activadoras de la 5-Lipooxigenasa , Enfermedad Aguda , Administración Oral , Animales , Antiinflamatorios no Esteroideos/uso terapéutico , Asma/tratamiento farmacológico , Asma/enzimología , Asma/metabolismo , Enfermedad Crónica , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Extravasación de Materiales Terapéuticos y Diagnósticos/tratamiento farmacológico , Extravasación de Materiales Terapéuticos y Diagnósticos/enzimología , Extravasación de Materiales Terapéuticos y Diagnósticos/metabolismo , Femenino , Humanos , Indoles/uso terapéutico , Inflamación/enzimología , Inflamación/metabolismo , Leucotrieno B4/biosíntesis , Leucotrieno B4/sangre , Masculino , Ratones , Ratones Endogámicos BALB C , Neumonía/tratamiento farmacológico , Neumonía/enzimología , Neumonía/metabolismo , Propionatos/uso terapéutico , Ratas , Ratas Sprague-Dawley , Zimosan

RESUMEN

The synthesis of a series of tricyclic antagonists for the prostaglandin D(2) receptor DP2 (CRTH2) is disclosed. The activities of the compounds were evaluated in a human DP2 binding assay and a human whole blood eosinophil shape change assay. Potential metabolic liabilities of the compounds were addressed through in vitro CYP studies. The lead compound was demonstrated to have efficacy in a mouse model of allergic rhinitis following oral dosing.

Asunto(s)

Antialérgicos/química , Antiinflamatorios/química , Compuestos Heterocíclicos con 3 Anillos/química , Receptores Inmunológicos/antagonistas & inhibidores , Receptores de Prostaglandina/antagonistas & inhibidores , Rinitis Alérgica Perenne/tratamiento farmacológico , Animales , Antialérgicos/síntesis química , Antialérgicos/farmacología , Antiinflamatorios/síntesis química , Antiinflamatorios/farmacología , Sistema Enzimático del Citocromo P-450/metabolismo , Modelos Animales de Enfermedad , Femenino , Compuestos Heterocíclicos con 3 Anillos/síntesis química , Compuestos Heterocíclicos con 3 Anillos/farmacología , Humanos , Ratones , Receptores Inmunológicos/metabolismo , Receptores de Prostaglandina/metabolismo

RESUMEN

The present study used an elevated platform procedure to investigate the effects of diazepam, a CRF1 antagonist CP-154,526 and a group II mGlu2/3 receptor agonist LY379268 on stress-evoked increase in extracellular norepinephrine (NE). Pretreatment with either diazepam (1 mg/kg, i.p.), CP-154,526 (20 mg/kg, i.p.) or LY379268 (1, 3 and 10 mg/kg, p.o.) significantly reduced platform stress-evoked NE. Interestingly, at the highest dose tested (10 mg/kg) LY379268 caused a marked increase in baseline NE levels. We tested whether this effect would diminish after repeated dosing. In contrast to acute administration, a challenge injection of LY379268 after repeated dosing (10 mg/kg x days) did not alter basal NE. Importantly, although less effective, LY379268 still significantly reduced stress-evoked NE. We further show that this increase in basal NE may involve mGlu2/3 receptor regulation of the GABAergic system. To this end, administration of the GABAB agonist, baclofen (4 mg/kg, i.p.), 2 h after dosing with LY379268, reversed the increase in baseline NE. These data suggest that, like diazepam and CP-154,526, group II mGlu2/3 receptor agonists can attenuate stress-evoked increase in extracellular NE in the rat prefrontal cortex. In addition they reveal a 'stress-like' increase in NE after high doses of LY379268 which may reflect mGlu3 receptor modulation of GABAergic transmission.

Asunto(s)

Aminoácidos/farmacología , Compuestos Bicíclicos Heterocíclicos con Puentes/farmacología , Diazepam/farmacología , Norepinefrina/metabolismo , Pirimidinas/farmacología , Pirroles/farmacología , Receptores de Hormona Liberadora de Corticotropina/antagonistas & inhibidores , Receptores de Glutamato Metabotrópico/agonistas , Animales , Líquido Extracelular/efectos de los fármacos , Líquido Extracelular/metabolismo , Masculino , Ratas , Ratas Sprague-Dawley , Receptores de Hormona Liberadora de Corticotropina/metabolismo , Receptores de Glutamato Metabotrópico/metabolismo , Estrés Fisiológico/metabolismo

RESUMEN

Herein we disclose the discovery of a new class of positive allosteric potentiators of the metabotropic glutamate receptor 2 (mGlu2), phenyl-tetrazolyl acetophenones, e.g. 1-(2-hydroxy-3-propyl-4-[4-[4-(2H-tetrazol-5-yl)phenoxy]butoxy]phenyl) ethanone (4). These potentiators were shown to have no effect in the absence of glutamate as well as no effect at mGlu3 or the other mGlu receptors. The compounds were also evaluated in rodent models with potential relevance for schizophrenia, and 4 was shown to have activity in the inhibition of ketamine-induced norepinephrine release and ketamine-induced hyperactivity. This represents the first example of the efficacy of mGlu2 receptor potentiators in these models.

Asunto(s)

Acetofenonas/síntesis química , Acetofenonas/farmacología , Regulación Alostérica , Receptores de Glutamato Metabotrópico/agonistas , Animales , Conducta/efectos de los fármacos , Encéfalo/metabolismo , Relación Dosis-Respuesta a Droga , Ácido Glutámico/farmacología , Hipercinesia/tratamiento farmacológico , Norepinefrina/metabolismo , Ratas , Esquizofrenia/tratamiento farmacológico , Relación Estructura-Actividad