Evaluation of osteogenic effects of BMP2 up-regulator E40071 in vitro / 药学学报

ABSTRACT

Bone morphogenetic protein 2 (BMP2) plays a key role in bone development and reestablishment. In the study, we screened up-regulators of BMP2 among 20 000 compounds through a cell-based high throughput screening model and a positive compound E40071[2-(4-(5-methyl-3-phenylpyrazolo[1,5-a]pyrimidin-7-yl) piperazin-1-yl)ethan-1-ol] was found as the positive hit. The EC50 value of E40071 was 2.73 μmol·L-1. In vitro, E40071 upregulated the mRNA levels of BMP2 and the downstream transcription factors, Runx2 and Osx in MC3T3-E1 (subclone 14). Protein expression of Runx2 was up-regulated by E40071 through induction of Smad1/5/8 phosphorylation. The alkaline phosphatase (ALP) activity was increased by E40071. Moreover, E40071 promoted the mineralization of MC3T3-E1 (subclone 14) by Alizarin red S staining. In addition, E40071 markedly inhibited osteoclast differentiation of mice macrophage Raw264.7 induced by RANKL and reduced the expression of osteoclast differentiation markers, including MMP9 and NFATc1. The results suggest that E40071 is able to promote bone formation activity of osteoblasts and inhibit differentiation of osteoclasts.