RESUMO
A novel class of mitogen-activated protein kinase-activated protein kinase 2 (MAPKAP-K2) inhibitors was discovered through screening a kinase-focused library. A homology model of MAPKAP-K2 was generated and used to guide the initial SAR studies and to rationalize the observed selectivity over CDK2. An X-ray crystal structure of a compound from the active series bound to crystalline MAPKAP-K2 confirmed the predicted binding mode. This has enabled the discovery of a series of pyrazolo[1,5-a]pyrimidine derivatives showing good in vitro cellular potency as anti-TNF-α agents and in vivo efficacy in a mouse model of endotoxin shock.
Assuntos
Anti-Inflamatórios não Esteroides/síntese química , Peptídeos e Proteínas de Sinalização Intracelular/antagonistas & inibidores , Proteínas Serina-Treonina Quinases/antagonistas & inibidores , Pirazóis/síntese química , Pirimidinas/síntese química , Animais , Anti-Inflamatórios não Esteroides/farmacocinética , Anti-Inflamatórios não Esteroides/farmacologia , Linhagem Celular , Cristalografia por Raios X , Proteínas de Choque Térmico HSP27/metabolismo , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/química , Lipopolissacarídeos/farmacologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Modelos Moleculares , Fosforilação , Conformação Proteica , Proteínas Serina-Treonina Quinases/química , Pirazóis/farmacocinética , Pirazóis/farmacologia , Pirimidinas/farmacocinética , Pirimidinas/farmacologia , Choque Séptico/metabolismo , Bibliotecas de Moléculas Pequenas , Estereoisomerismo , Relação Estrutura-Atividade , Fator de Necrose Tumoral alfa/biossínteseRESUMO
A sensitive HPLC method for simultaneous determination of endogenous levels of all-trans-(ATRA), 13-cis-(13cRA), and 9-cis-retinoic acids (9cRA) was applied to serum samples from healthy volunteers and type II diabetes mellitus patients. Levels of 9cRA (around 0.2 ng/ml in both groups) were below the limit of quantification. The concentrations of ATRA and 13cRA were reliably quantified, and the within-day and between-days variances indicated that they were well maintained with little variation. Concentrations of serum ATRA and 13cRA of diabetic patients (ATRA: 1.76 +/- 0.54 ng/ml; 13cRA: 1.77 +/- 0.39ng/ml, n=13) were rather lower than those of healthy subjects (ATRA: 2.08 +/- 0.53 ng/ml; 13cRA: 2.05 +/- 0.26 ng/ml, n = 18), but the differences were not significant, except for the sum of ATRA and 13cRA (p = 0.033). Interestingly, the serum levels of retinoic acids in diabetic patients correlated positively with the hemoglobin A1c (HbA1c) values (ATRA: r = 0.57, p < 0.05; 13cRA: r = 0.62, p < 0.05). The results warrant further studies on the possible involvement of uncontrolled serum retinoic acids levels in the pathogenesis and/or treatment of diabetes mellitus.
