RESUMEN
The connection between Netherton syndrome and overactivation of epidermal/dermal proteases, particularly Kallikrein 5 (KLK5) has been well established and it is expected that a KLK5 inhibitor would improve the dermal barrier and also reduce the pain and itch that afflict Netherton syndrome patients. One of the challenges of covalent protease inhibitors has been achieving selectivity over closely related targets. In this paper we describe the use of structural insight to design and develop a selective and highly potent reversibly covalent KLK5 inhibitor from an initial weakly binding fragment.
Asunto(s)
Benzamidinas/química , Calicreínas/antagonistas & inhibidores , Síndrome de Netherton/tratamiento farmacológico , Inhibidores de Serina Proteinasa/química , Secuencia de Aminoácidos , Benzamidinas/farmacología , Sitios de Unión , Evaluación Preclínica de Medicamentos , Humanos , Isomerismo , Modelos Moleculares , Estructura Molecular , Mutación , Unión Proteica , Inhibidor de Serinpeptidasas Tipo Kazal-5/genética , Inhibidores de Serina Proteinasa/farmacología , Relación Estructura-ActividadRESUMEN
Regulation of proteolytic activity in the skin plays a pivotal role in epidermal homeostasis. This is best exemplified in Netherton syndrome, a severe genetic skin condition caused by loss-of-function mutations in the gene serine protease inhibitor Kazal-type 5 encoding lympho-epithelial Kazal-type-related inhibitor, a serine protease inhibitor that regulates kallikrein (KLK)-related peptidase 5, 7, and 14 activities. KLK5 plays a central role in stratum corneum shedding and inflammatory cell signaling, activates KLK7 and KLK14, and is therefore an optimal therapeutic target. We aimed to identify a potent and selective small-molecule inhibitor of KLK5 amenable to epidermal delivery. GSK951 was identified using a structure-based design strategy and showed a half maximal inhibitory concentration of 250 pM for KLK5 and greater than 100-fold selectivity over KLK7 and KLK14. Cocrystal structure analysis identified the critical catalytic site interactions to a surrogate for KLK5. Topical application of GSK951-containing cream inhibited KLK5 activity in TgKLK5 mouse skin, reduced transepidermal water loss, and decreased proinflammatory cytokine expression. GSK951 achieved high concentrations in healthy human epidermis following topical application in a cream formulation. Finally, KLK5 protease activity was increased in stratum corneum of patients with Netherton syndrome and significantly inhibited by GSK951. These findings unveil a KLK5-specific small-molecule inhibitor with a high therapeutic potential for patients with Netherton syndrome.
Asunto(s)
Antiinflamatorios/uso terapéutico , Compuestos de Boro/uso terapéutico , Inflamación/tratamiento farmacológico , Calicreínas/antagonistas & inhibidores , Síndrome de Netherton/tratamiento farmacológico , Piel/patología , Administración Tópica , Animales , Modelos Animales de Enfermedad , Humanos , Calicreínas/genética , Ratones , Ratones Transgénicos , Transducción de Señal , Piel/efectos de los fármacos , Crema para la PielRESUMEN
Starting from the structure of Telmisartan, a new series of potent and selective PPARgamma modulators was identified. The synthesis, in vitro and in vivo evaluation of the most potent compounds are reported and the X-ray structure of compound 7b bound to the PPARgamma ligand binding domain is described.
Asunto(s)
Diseño de Fármacos , Indoles/síntesis química , PPAR gamma/efectos de los fármacos , Inhibidores de la Enzima Convertidora de Angiotensina/química , Inhibidores de la Enzima Convertidora de Angiotensina/farmacología , Animales , Bencimidazoles/química , Bencimidazoles/farmacología , Benzoatos/química , Benzoatos/farmacología , Cristalografía por Rayos X , Humanos , Indoles/química , Indoles/farmacología , Concentración 50 Inhibidora , Modelos Moleculares , Estructura Molecular , Ratas , Relación Estructura-Actividad , TelmisartánRESUMEN
Starting from the structure of 5, a two-step strategy was applied to identify a new generation of trifluoromethane sulfonamides as potent PPARalpha agonists. Synthesis, in vitro and in vivo evaluation of the most potent compound are reported.
Asunto(s)
PPAR alfa/agonistas , Sulfonamidas/química , Sulfonamidas/farmacología , Animales , Línea Celular , Diseño de Fármacos , Evaluación Preclínica de Medicamentos , Humanos , Ratones , Sulfonamidas/síntesis químicaRESUMEN
We report the design and synthesis of equipotent PPARalpha/gamma dual agonists starting from selective PPAR alpha agonist 1. In vivo data for 7 in the Zucker fa/fa rat are presented.
Asunto(s)
PPAR alfa/agonistas , PPAR gamma/agonistas , Pirazoles/síntesis química , Animales , Técnicas Químicas Combinatorias , Diseño de Fármacos , Estructura Molecular , Isoformas de Proteínas , Pirazoles/química , Pirazoles/toxicidad , Ratas , Ratas Zucker , Relación Estructura-ActividadRESUMEN
The peroxisome proliferator activated receptors PPARalpha, PPARgamma, and PPARdelta are ligand-activated transcription factors that play a key role in lipid homeostasis. The fibrates raise circulating levels of high-density lipoprotein cholesterol and lower levels of triglycerides in part through their activity as PPARalpha agonists; however, the low potency and restricted selectivity of the fibrates may limit their efficacy, and it would be desirable to develop more potent and selective PPARalpha agonists. Modification of the selective PPARdelta agonist 1 (GW501516) so as to incorporate the 2-aryl-2-methylpropionic acid group of the fibrates led to a marked shift in potency and selectivity toward PPARalpha agonism. Optimization of the series gave 25a, which shows EC50 = 4 nM on PPARalpha and at least 500-fold selectivity versus PPARdelta and PPARgamma. Compound 25a (GW590735) has been progressed to clinical trials for the treatment of diseases of lipid imbalance.
Asunto(s)
HDL-Colesterol/sangre , PPAR alfa/agonistas , Propionatos/síntesis química , Tiazoles/síntesis química , Animales , Apolipoproteína A-I/genética , VLDL-Colesterol/sangre , Cristalografía por Rayos X , Perros , Dislipidemias/sangre , Dislipidemias/tratamiento farmacológico , Humanos , Ligandos , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Modelos Moleculares , PPAR alfa/química , Propionatos/farmacocinética , Propionatos/farmacología , Estructura Terciaria de Proteína , Ratas , Ratas Wistar , Relación Estructura-Actividad , Tiazoles/farmacocinética , Tiazoles/farmacología , Triglicéridos/sangreRESUMEN
1 Chronic liver disease is characterized by an exacerbated accumulation of matrix, causing progressive fibrosis, which may lead to cirrhosis. Transforming growth factor beta (TGF-beta), a well-known profibrotic cytokine, transduces its signal through the ALK5 ser/thr kinase receptor, and increases transcription of different genes including PAI-1 and collagens. The identification of GW6604 (2-phenyl-4-(3-pyridin-2-yl-1H-pyrazol-4-yl)pyridine), an ALK5 inhibitor, allowed us to evaluate the therapeutic potential of inhibiting TGF-beta pathway in different models of liver disease. 2 A cellular assay was used to identify GW6604 as a TGF-beta signaling pathway inhibitor. This ALK5 inhibitor was then tested in a model of liver hepatectomy in TGF-beta-overexpressing transgenic mice, in an acute model of liver disease and in a chronic model of dimethylnitrosamine (DMN)-induced liver fibrosis. 3 In vitro, GW6604 inhibited autophosphorylation of ALK5 with an IC(50) of 140 nM and in a cellular assay inhibited TGF-beta-induced transcription of PAI-1 (IC(50): 500 nM). In vivo, GW6604 (40 mg kg(-1) p.o.) increased liver regeneration in TGF-beta-overexpressing mice, which had undergone partial hepatectomy. In an acute model of liver disease, GW6604 reduced by 80% the expression of collagen IA1. In a chronic model of DMN-induced fibrosis where DMN was administered for 6 weeks and GW6604 dosed for the last 3 weeks (80 mg kg(-1) p.o., b.i.d.), mortality was prevented and DMN-induced elevations of mRNA encoding for collagen IA1, IA2, III, TIMP-1 and TGF-beta were reduced by 50-75%. Inhibition of matrix genes overexpression was accompanied by reduced matrix deposition and reduction in liver function deterioration, as assessed by bilirubin and liver enzyme levels. 4 Our results suggest that inhibition of ALK5 could be an attractive new approach to treatment of liver fibrotic diseases by both preventing matrix deposition and promoting hepatocyte regeneration.
Asunto(s)
Receptores de Activinas Tipo I/antagonistas & inhibidores , Dimetilnitrosamina , Cirrosis Hepática/prevención & control , Pirazoles/farmacología , Piridinas/farmacología , Receptores de Factores de Crecimiento Transformadores beta/antagonistas & inhibidores , Factor de Crecimiento Transformador beta/antagonistas & inhibidores , Receptores de Activinas Tipo I/metabolismo , Enfermedad Aguda , Animales , Línea Celular Tumoral , Enfermedad Crónica , Hepatectomía , Humanos , Cirrosis Hepática/inducido químicamente , Cirrosis Hepática/patología , Regeneración Hepática/efectos de los fármacos , Masculino , Ratones , Ratones Transgénicos , Fosforilación , Inhibidor 1 de Activador Plasminogénico/biosíntesis , Inhibidor 1 de Activador Plasminogénico/genética , Proteínas Serina-Treonina Quinasas , Ratas , Ratas Sprague-Dawley , Receptor Tipo I de Factor de Crecimiento Transformador beta , Receptores de Factores de Crecimiento Transformadores beta/metabolismo , Transducción de Señal , Transcripción Genética , Factor de Crecimiento Transformador beta/genética , Factor de Crecimiento Transformador beta/fisiologíaRESUMEN
Optimization of the screening hit 1 led to the identification of novel 1,5-naphthyridine aminothiazole and pyrazole derivatives, which are potent and selective inhibitors of the transforming growth factor-beta type I receptor, ALK5. Compounds 15 and 19, which inhibited ALK5 autophosphorylation with IC50 = 6 and 4 nM, respectively, showed potent activities in both binding and cellular assays and exhibited selectivity over p38 mitogen-activated protein kinase. The X-ray crystal structure of 19 in complex with human ALK5 is described, confirming the binding mode proposed from docking studies.