RESUMO
A novel series of trisubstituted ureas has been identified as potent and selective mPGES-1 inhibitors. These compounds are selective over other prostanoid enzymes such as PGF synthase and TX synthase. This series of inhibitors was developed by lead optimization of a hit from an internal HTS campaign. Lead compound 42 is potent in A549 cell assay (IC(50) of 0.34 µM) and in human whole blood assay (IC(50) of 2.1 µM). An efficient and versatile one-pot strategy for the formation of ureas, involving a reductive amination, was developed to generate these inhibitors.
Assuntos
Oxirredutases Intramoleculares/antagonistas & inibidores , Ureia/síntese química , Linhagem Celular Tumoral , Humanos , Microssomos/enzimologia , Prostaglandina-E Sintases , Relação Estrutura-Atividade , Ureia/química , Ureia/farmacologiaRESUMO
Phenanthrene imidazoles 26 and 44 have been identified as novel potent, selective and orally active mPGES-1 inhibitors. These inhibitors are significantly more potent than the previously reported chlorophenanthrene imidazole 1 (MF63) with a human whole blood IC50 of 0.20 and 0.14 microM, respectively. It exhibited a significant analgesic effect in a guinea pig hyperalgesia model at oral doses as low as 14 mg/kg. Both active and selective mPGES-1 inhibitors (26 and 44) have a relatively distinct pharmacokinetic profile and are suitable for clinical development.
Assuntos
Benzimidazóis/química , Inibidores Enzimáticos/química , Oxirredutases Intramoleculares/antagonistas & inibidores , Nitrilas/química , Fenantrenos/química , Administração Oral , Animais , Benzimidazóis/síntese química , Benzimidazóis/farmacocinética , Modelos Animais de Doenças , Desenho de Fármacos , Inibidores Enzimáticos/síntese química , Inibidores Enzimáticos/farmacocinética , Cobaias , Humanos , Hiperalgesia/tratamento farmacológico , Oxirredutases Intramoleculares/metabolismo , Nitrilas/síntese química , Nitrilas/farmacocinética , Fenantrenos/síntese química , Fenantrenos/farmacocinética , Prostaglandina-E Sintases , Proteínas Recombinantes/antagonistas & inibidores , Proteínas Recombinantes/metabolismo , Relação Estrutura-AtividadeRESUMO
A SAR study of a series of 1-phenyl-1,8-naphthyridin-4-one-3-carboxamides is described. Optimization of the series was based on in vitro potency against PDE4, inhibition of the LPS-induced production of TNF-alpha in human whole blood and minimizing affinity for the hERG potassium channel. From these studies, compounds 18 and 20 (MK-0873) were identified as optimized PDE4 inhibitors with good overall in vitro and in vivo profiles and selected as development candidates.
Assuntos
Química Farmacêutica/métodos , Nucleotídeo Cíclico Fosfodiesterase do Tipo 4/química , Naftiridinas/síntese química , Animais , Cães , Desenho de Fármacos , Canal de Potássio ERG1 , Canais de Potássio Éter-A-Go-Go/química , Humanos , Concentração Inibidora 50 , Lipopolissacarídeos/química , Modelos Químicos , Naftiridinas/farmacologia , Ligação Proteica , Ratos , Saimiri , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Phenanthrene imidazole 3 (MF63) has been identified as a novel potent, selective, and orally active mPGES-1 inhibitor. This new series was developed by lead optimization of a hit from an internal HTS campaign. Compound 3 is significantly more potent than the previously reported indole carboxylic acid 1 with an A549 whole cell IC(50) of 0.42 microM (50% FBS) and a human whole blood IC(50) of 1.3 microM. It exhibited a significant analgesic effect in a guinea pig hyperalgesia model when orally dosed at 30 and 100mg/kg.
Assuntos
Analgésicos não Narcóticos/síntese química , Imidazóis/síntese química , Imidazóis/farmacologia , Oxirredutases Intramoleculares/antagonistas & inibidores , Fenantrenos/síntese química , Fenantrenos/farmacologia , Analgésicos não Narcóticos/sangue , Analgésicos não Narcóticos/química , Analgésicos não Narcóticos/farmacologia , Animais , Modelos Animais de Doenças , Desenho de Fármacos , Cobaias , Humanos , Hiperalgesia/induzido quimicamente , Imidazóis/sangue , Imidazóis/química , Concentração Inibidora 50 , Estrutura Molecular , Fenantrenos/sangue , Fenantrenos/química , Prostaglandina-E Sintases , Ratos , Relação Estrutura-AtividadeRESUMO
A SAR study on the tertiary alcohol series of phosphodiesterase-4 (PDE4) inhibitors related to 1 is described. In addition to inhibitory potency against PDE4 and the lipopolysaccharide-induced production of TNFalpha in human whole blood, the binding affinity of these compounds for the human ether-a-go-go related gene (hERG) potassium channel (an in vitro measure for the potential to cause QTc prolongation) was assessed. Four key structural moieties in the molecule were studied, and the impact of the resulting modifications in modulating these activities was evaluated. From these studies, (+)-3d (L-869,298) was identified as an optimized structure with respect to PDE4 inhibitory potency, lack of binding affinity to the hERG potassium channel, and pharmacokinetic behavior. (+)-3d exhibited good in vivo efficacy in several models of pulmonary function with a wide therapeutic index with respect to emesis and prolongation of the QTc interval.
Assuntos
3',5'-AMP Cíclico Fosfodiesterases/antagonistas & inibidores , Álcoois/síntese química , Óxidos N-Cíclicos/síntese química , Inibidores de Fosfodiesterase/síntese química , Canais de Potássio de Abertura Dependente da Tensão da Membrana , Canais de Potássio/metabolismo , Piridinas/síntese química , Álcoois/farmacocinética , Álcoois/farmacologia , Álcoois/toxicidade , Animais , Broncoconstrição/efeitos dos fármacos , Cristalografia por Raios X , Óxidos N-Cíclicos/farmacocinética , Óxidos N-Cíclicos/farmacologia , Óxidos N-Cíclicos/toxicidade , Nucleotídeo Cíclico Fosfodiesterase do Tipo 4 , Cães , Canal de Potássio ERG1 , Eletrocardiografia , Canais de Potássio Éter-A-Go-Go , Cobaias , Humanos , Técnicas In Vitro , Síndrome do QT Longo/induzido quimicamente , Inibidores de Fosfodiesterase/farmacocinética , Inibidores de Fosfodiesterase/farmacologia , Inibidores de Fosfodiesterase/toxicidade , Ligação Proteica , Piridinas/química , Piridinas/farmacocinética , Piridinas/farmacologia , Piridinas/toxicidade , Ratos , Saimiri , Ovinos , Estereoisomerismo , Relação Estrutura-Atividade , Fator de Necrose Tumoral alfa/biossíntese , Vômito/induzido quimicamenteRESUMO
Phosphodiesterase 4 (PDE4) inhibitors elevate cyclic adenosine 5'-monophosphate (cAMP), and this elevation has been shown to inhibit inflammatory cytokines such as tumor necrosis factor-alpha (TNF-alpha). Using TNF-alpha as a biomarker, we have developed transcription-based assays to examine inhibition of PDE4 activity in human and guinea pig whole blood. In vitro inhibition by PDE4 inhibitors was measured using quantitative PCR (qPCR) analysis of TNF-alpha mRNA levels in whole blood stimulated with lipopolysaccharide (LPS). The kinetics of human TNF-alpha mRNA production were analyzed and shown to be highest 4 hr following LPS stimulation. The guinea pig displayed kinetics of TNF-alpha transcription similar to those of the human. Analysis of inhibition of human TNF-alpha protein production was performed by immunoassay and shown to correlate with inhibition of transcription for three of the four compounds tested. Roflumilast was found to be 9-fold more potent for TNF-alpha inhibition in the qPCR assay than in the protein assay. The potencies of L-826,141 and roflumilast were determined in human and guinea pig whole blood by qPCR, with IC(50) values of 270 and 20 nM, respectively, in humans and 100 and 10 nM, respectively, in guinea pigs. These results show that the potency of PDE4 inhibitors can be monitored in whole blood using a transcription-based assay, and that this type of assay can be adapted to various species provided the TNF-alpha nucleotide sequence is known. The in vitro whole blood IC(50) for TNF-alpha inhibition was compared to inhibition in the ovalbumin-challenged guinea pig model of bronchoconstriction. Obtaining plasma levels at the IC(50) determined in vitro for L-826,141 and roflumilast provides significant inhibition of bronchoconstriction. This suggests that TNF-alpha can be used as a whole blood biomarker in the guinea pig for PDE4 inhibition in this inflammatory model.
Assuntos
3',5'-AMP Cíclico Fosfodiesterases/metabolismo , Broncoconstrição/efeitos dos fármacos , Ovalbumina/farmacologia , Inibidores de Fosfodiesterase/farmacologia , Piridinas/farmacologia , Fator de Necrose Tumoral alfa/metabolismo , 3',5'-AMP Cíclico Fosfodiesterases/antagonistas & inibidores , Animais , Broncoconstrição/fisiologia , Nucleotídeo Cíclico Fosfodiesterase do Tipo 4 , Interações Medicamentosas , Feminino , Cobaias , Humanos , Lipopolissacarídeos/farmacologia , Masculino , Inibidores de Fosfodiesterase/química , Reação em Cadeia da Polimerase , Piridinas/química , RNA Mensageiro/sangue , RNA Mensageiro/efeitos dos fármacos , Fatores de Tempo , Fator de Necrose Tumoral alfa/genéticaRESUMO
OBJECTIVE: The objectives of our study were to compare patient characteristics and severity of disease in obese and normal-weight-confirmed people with asthma and to explore reasons for misdiagnosis of asthma, including potential interactions with obesity. METHODS: We randomly selected patients with physician-diagnosed asthma from eight Canadian cities. Asthma diagnosis was confirmed via a sequential lung function testing algorithm. Logistic analysis was conducted to compare obese and normal-weight-confirmed people with asthma and to assess characteristics associated with misdiagnosis of asthma. Interaction with obesity was investigated. RESULTS: Complete assessments were obtained on 496 subjects who reported physician-diagnosed asthma (242 obese and 254 normal-weight subjects); 346 had asthma confirmed with sequential lung testing, and in 150 subjects a diagnosis of asthma was ruled out. Obese subjects with asthma were significantly more likely to be men, have a history of hypertension and gastroesophageal reflux disease, and have a lower FEV(1) compared with normal-weight subjects with asthma. Older subjects, men, and subjects with higher FEV(1) were more likely to have received misdiagnoses of asthma. Obesity was not an independent predictor of misdiagnosis, however there was an interaction between obesity and urgent visits for respiratory symptoms. The odds ratio for receiving a misdiagnosis of asthma for obese individuals as compared with normal-weight individuals was 4.08 (95% CI, 1.23-13.5) for those with urgent visits in the past 12 months. CONCLUSIONS: Obese people with asthma have lower lung function and more comorbidities compared with normal-weight people with asthma. Obese individuals who make urgent visits for respiratory symptoms are more likely to receive a misdiagnosis of asthma.
Assuntos
Asma/complicações , Asma/diagnóstico , Obesidade/complicações , Adulto , Fatores Etários , Algoritmos , Asma/epidemiologia , Asma/fisiopatologia , Índice de Massa Corporal , Canadá/epidemiologia , Distribuição de Qui-Quadrado , Feminino , Volume Expiratório Forçado , Refluxo Gastroesofágico/complicações , Refluxo Gastroesofágico/epidemiologia , Humanos , Hipertensão/complicações , Hipertensão/epidemiologia , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Obesidade/epidemiologia , Obesidade/fisiopatologia , Qualidade de Vida , Testes de Função Respiratória , Fatores de Risco , Índice de Gravidade de DoençaRESUMO
The discovery and SAR of a new series of substituted 8-arylquinoline PDE4 inhibitors are herein described. This work has led to the identification of several compounds with excellent in vitro and in vivo profiles, including a good therapeutic window of emesis to efficacy in several animal models. Typical optimized compounds from this series are potent inhibitors of PDE4 (IC(50)<1nM) and also of LPS-induced TNF-alpha release in human whole blood (IC(50)<0.5microM). The same compounds are potent inhibitors of ovalbumin-induced bronchoconstriction in conscious guinea pigs (EC(50)<0.1mg/kg ip) but require a dose of about 10mg/kg po in the squirrel monkey to produce an emetic response. From this series of compounds, 23a (L-454,560) was identified as an optimized compound.
Assuntos
3',5'-AMP Cíclico Fosfodiesterases/antagonistas & inibidores , Inibidores de Fosfodiesterase/química , Inibidores de Fosfodiesterase/farmacologia , Quinolinas/química , Quinolinas/farmacologia , Animais , Broncoconstrição/efeitos dos fármacos , Nucleotídeo Cíclico Fosfodiesterase do Tipo 4 , Cobaias , Humanos , Concentração Inibidora 50 , Inibidores de Fosfodiesterase/toxicidade , Quinolinas/toxicidade , Ratos , Saimiri , Ovinos , Relação Estrutura-Atividade , Vômito/induzido quimicamenteRESUMO
Microsomal prostaglandin E synthase-1 (mPGES-1) is an inducible protein recently shown to be an important source of inflammatory PGE2. Here we have used mPGES-1 wild type, heterozygote, and null mice to assess the impact of reduction or absence mPGES-1 protein on the production of PGE2 and other prostaglandins in lipopolysaccharide (LPS)-treated macrophages and mice. Thioglycollate-elicited peritoneal macrophages with mPGES-1 deficiency were found to lose their ability to produce PGE2 upon LPS stimulation. Resident mPGES-1(-/-) peritoneal macrophages exhibited severely impaired PGE2-releasing activity but retained some LPS-inducible PGE2 production capacity. Both macrophage types showed a 50% decrease in PGE2 production with removal of one copy of the mPGES-1 gene. In vivo, mPGES-1 deletion abolished the LPS-stimulated production of PGE2 in spleen, kidney, and brain. Surprisingly, lack of mPGES-1 activity resulted in an 80-90% decrease in basal, cyclooxygenase-1 (COX-1)-dependent PGE2 production in stomach and spleen, and a 50% reduction in brain and kidney. Other prostaglandins (thromboxane B2, PGD2, PGF(2alpha), and 6-keto-PGF(1alpha)) were significantly elevated in stomachs of mPGES-1-null mice but not in other tissues. Examination of mRNA for several terminal prostaglandin synthases did not reveal changes in expression levels associated with mPGES-1 deficiency, indicating that gastric prostaglandin changes may be due to shunting of cyclooxygenase products to other terminal synthases. These data demonstrate for the first time a dual role for mPGES-1 in both inflammatory and COX-1-mediated PGE2 production and suggest an interdependence of prostanoid production with tissue-specific alterations of prostaglandin levels in the absence of mPGES-1.