RESUMEN
This Letter describes methodology to enable the identification of tool or therapeutic lipopeptides which modulate the function of membrane bound proteins. The choice of lipopeptides as a chemotype is the amalgamation of multiple medicinal chemistry considerations including duration of action, low systemic exposure and access to intracellular components. The 'lipopeptide shuffle' has been applied here to the APJ receptor and has rapidly resulted in the discovery of a 33 nM APJ agonist hit from an initial 369 member lipopeptide synthetic array.
Asunto(s)
Diseño de Fármacos , Lipopéptidos/farmacología , Receptores Acoplados a Proteínas G/agonistas , Receptores de Apelina , Relación Dosis-Respuesta a Droga , Humanos , Lipopéptidos/química , Lipopéptidos/genética , Conformación Molecular , Relación Estructura-ActividadRESUMEN
By considering published structural information we have designed high throughput biaryl lipophilic acid arrays leveraging facile chemistry to expedite their synthesis. We rapidly identified multiple hits which were of suitable IP agonist potency. These relatively simple and strategically undecorated molecules present an ideal opportunity for optimization towards our target candidate profile.
Asunto(s)
Antihipertensivos/química , Antihipertensivos/farmacología , Compuestos Bicíclicos con Puentes/química , Compuestos Bicíclicos con Puentes/farmacología , Receptores de Epoprostenol/agonistas , Ensayos Analíticos de Alto Rendimiento/métodos , Ligandos , Receptores de Epoprostenol/química , Relación Estructura-ActividadRESUMEN
A high-throughput screen targeting the EP(1) receptor identified non-acidic glycine sulfonamide derivative 2a with a pK(i) of 6.2. Analogue synthesis allowed a thorough investigation of the structure-activity relationship (SAR) and led to a 100-fold increase in recombinant potency.
Asunto(s)
Glicinérgicos/síntesis química , Glicinérgicos/farmacología , Receptores de Prostaglandina E/antagonistas & inhibidores , Sulfonamidas/síntesis química , Sulfonamidas/farmacología , Animales , Células CHO , Membrana Celular/metabolismo , Cromatografía Líquida de Alta Presión , Cricetinae , Cricetulus , Sistema Enzimático del Citocromo P-450/metabolismo , Dinoprostona/metabolismo , Evaluación Preclínica de Medicamentos , Humanos , Indicadores y Reactivos , Subtipo EP1 de Receptores de Prostaglandina ERESUMEN
The discovery, synthesis and structure-activity relationship (SAR) of novel carboxylic acid agonists for GPR40 are described. Aryl propionic acid 1, identified from a high throughput screen, was selected for chemical exploration. Compound 2 was identified as our lead molecule through efficient solid phase combinatorial array chemistry and had an attractive in vitro and in vivo pharmacokinetic profile in rat. These ligands may prove useful in establishing a role for GPR40 in insulin regulation.
Asunto(s)
Ácidos Carboxílicos/síntesis química , Ácidos Carboxílicos/farmacología , Receptores Acoplados a Proteínas G/agonistas , Animales , Disponibilidad Biológica , Células CHO , Ácidos Carboxílicos/farmacocinética , Fenómenos Químicos , Química Física , Cricetinae , Cricetulus , Evaluación Preclínica de Medicamentos , Indicadores y Reactivos , Ligandos , Unión Proteica , Ratas , Relación Estructura-ActividadRESUMEN
The discovery, synthesis and structure-activity relationship (SAR) of a novel series of EP1 receptor antagonists is described. Pyrazole acid 4, identified from a chemical array, had desirable physicochemical properties, an excellent in vitro microsomal inhibition and cytochrome P450 (CYP450) profile and good exposure levels in blood. This compound had an ED50 of 1.3 mg/kg in a rat pain model. A range of more potent analogues in the in vitro assay was identified using efficient array chemistry. These EP1 antagonists have potential as agents in the treatment of PGE2 mediated pain.
Asunto(s)
Pirazoles/síntesis química , Pirazoles/farmacología , Receptores de Prostaglandina E/antagonistas & inhibidores , Animales , Fenómenos Químicos , Química Física , Sistema Enzimático del Citocromo P-450/metabolismo , Estructura Molecular , Pirazoles/química , Pirazoles/farmacocinética , Ratas , Receptores de Prostaglandina E/metabolismo , Subtipo EP1 de Receptores de Prostaglandina E , Relación Estructura-ActividadRESUMEN
1. Long chain fatty acids have recently been identified as agonists for the G protein-coupled receptors GPR40 and GPR120. Here, we present the first description of GW9508, a small-molecule agonist of the fatty acid receptors GPR40 and GPR120. In addition, we also describe the pharmacology of GW1100, a selective GPR40 antagonist. These molecules were used to further investigate the role of GPR40 in glucose-stimulated insulin secretion in the MIN6 mouse pancreatic beta-cell line. 2. GW9508 and linoleic acid both stimulated intracellular Ca2+ mobilization in human embryonic kidney (HEK)293 cells expressing GPR40 (pEC50 values of 7.32+/-0.03 and 5.65+/-0.06, respectively) or GPR120 (pEC50 values of 5.46+/-0.09 and 5.89+/-0.04, respectively), but not in the parent HEK-293 cell line. 3. GW1100 dose dependently inhibited GPR40-mediated Ca2+ elevations stimulated by GW9508 and linoleic acid (pIC50 values of 5.99+/-0.03 and 5.99+/-0.06, respectively). GW1100 had no effect on the GPR120-mediated stimulation of intracellular Ca2+ release produced by either GW9508 or linoleic acid. 4. GW9508 dose dependently potentiated glucose-stimulated insulin secretion in MIN6 cells, but not in primary rat or mouse islets. Furthermore, GW9508 was able to potentiate the KCl-mediated increase in insulin secretion in MIN6 cells. The effects of GW9508 on insulin secretion were reversed by GW1100, while linoleic acid-stimulated insulin secretion was partially attenuated by GW1100. 5. These results add further evidence to a link between GPR40 and the ability of fatty acids to acutely potentiate insulin secretion and demonstrate that small-molecule GPR40 agonists are glucose-sensitive insulin secretagogues.
Asunto(s)
Células Secretoras de Insulina/citología , Células Secretoras de Insulina/efectos de los fármacos , Células Secretoras de Insulina/metabolismo , Insulina/metabolismo , Receptores Acoplados a Proteínas G/agonistas , Receptores Acoplados a Proteínas G/antagonistas & inhibidores , Animales , Benzoatos/farmacología , Células CHO , Línea Celular , Células Cultivadas , Cricetinae , Evaluación Preclínica de Medicamentos , Glucosa/farmacología , Humanos , Hipoglucemiantes/farmacología , Hipolipemiantes/farmacología , Secreción de Insulina , Metilaminas/farmacología , Ratones , Modelos Biológicos , Cloruro de Potasio/farmacología , Propionatos/farmacología , Pirimidinas/farmacología , Spodoptera/citologíaRESUMEN
A method is described whereby stable isotopic signatures were partially incorporated into both termini of a peptide sequence giving rise to a characteristic cluster of four peaks in the mass spectral analysis. Cleavage of this peptide by a protease between the labeled positions generates two fragments both displaying their own individual signature peaks. The event of protease cleavage of the peptide was monitored by the changes in clusters within the spectrum. We believe that this technique could be used to aid the discovery of new cleavage substrates for proteases. Additionally, the analysis can be automated with dedicated software designed to select and interpret the data since all peaks of interest contain predefined signatures and can be easily distinguished from background noise.