RESUMO
Hypothalamic CAMKK2 represents a potential mechanism for chemically affecting satiety and promoting weight loss in clinically obese patients. Single-digit nanomolar inhibitors of CAMKK2 were identified in three related ATP-competitive series. Limited optimization of kinase selectivity, solubility, and pharmacokinetic properties were undertaken on all three series, as SAR was often transferrable. Ultimately, a 2,4-diaryl 7-azaindole was optimized to afford a tool molecule that potently inhibits AMPK phosphorylation in a hypothalamus-derived cell line, is orally bioavailable, and crosses the blood-brain barrier. When dosed orally in rodents, compound 4â¯t limited ghrelin-induced food intake.
Assuntos
Quinase da Proteína Quinase Dependente de Cálcio-Calmodulina/antagonistas & inibidores , Inibidores de Proteínas Quinases/química , Administração Oral , Animais , Encéfalo/metabolismo , Quinase da Proteína Quinase Dependente de Cálcio-Calmodulina/genética , Quinase da Proteína Quinase Dependente de Cálcio-Calmodulina/metabolismo , Ingestão de Alimentos/efeitos dos fármacos , Grelina/farmacologia , Ligação de Hidrogênio , Indóis/química , Indóis/metabolismo , Concentração Inibidora 50 , Camundongos , Mutagênese , Inibidores de Proteínas Quinases/metabolismoRESUMO
In response to the increasing demands to generate larger amounts of quality data faster, specifically in the area of RNA isolation to support gene expression assays, the authors have adopted several automated solutions for isolating total RNA from a variety of sample types, (eg, blood, cells, and tissue). By using automated solutions, the authors were able to increase their throughput without compromising data integrity.
Assuntos
RNA/isolamento & purificação , Animais , Automação , Células/metabolismo , Humanos , Reação em Cadeia da Polimerase , Controle de Qualidade , RNA/sangue , Reprodutibilidade dos TestesRESUMO
Rosiglitazone was found to simulate mitochondrial biogenesis in mouse brain in an apolipoprotein (Apo) E isozyme-independent manner. Rosiglitazone induced both mitochondrial DNA (mtDNA) and estrogen-stimulated related receptor alpha (ESRRA) mRNA, a key regulator of mitochondrial biogenesis. Transcriptomics and proteomics analysis suggested the mitochondria produced in the presence of human ApoE3 and E4 were not as metabolically efficient as those in the wild type or ApoE knockout mice. Thus, we propose that PPARgamma agonism induces neuronal mitochondrial biogenesis and improves glucose utilization leading to improved cellular function and provides mechanistic support for the improvement in cognition observed in treatment of Alzheimer's patients with rosiglitazone.
