Here, we describe three new small-molecule activators of BMP signaling found by high throughput screening of a library of ∼600 000 small molecules. Using a cell-based luciferase assay in the BMP4-responsive human cervical carcinoma clonal cell line, C33A-2D2, we identified three compounds with similar chemotypes that each ventralize zebrafish embryos and stimulate increased expression of the BMP target genes, bmp2b and szl. Because these compounds ventralize zebrafish embryos, we have termed them “ventromorphins.” As expected for a BMP pathway activator, they induce the differentiation of C2C12 myoblasts to osteoblasts. Affymetrix RNA analysis confirmed the differentiation results and showed that ventromorphins treatment elicits a genetic response similar to BMP4 treatment. Unlike isoliquiritigenin (SJ000286237), a flavone that maximally activates the pathway after 24 h of treatment, all three ventromorphins induced SMAD1/5/8 phosphorylation within 30 min of treatment and achieved peak activity within 1 h, indicating that their responses are consistent with directly activating BMP signaling.

中文翻译：

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Ventromorphins：新型的典型BMP信号通路的小分子激活剂

在这里，我们描述了通过高通量筛选约60万个小分子文库发现的BMP信号转导的三个新小分子活化剂。使用基于细胞的萤光素酶测定法在BMP4反应性人宫颈癌克隆细胞系C33A-2D2中鉴定了三种具有相似化学型的化合物，每种化合物使斑马鱼胚胎腹侧并刺激BMP目标基因bmp2b和szl的表达增加。因为这些化合物使斑马鱼的胚胎腹侧化，所以我们将它们称为“ ventromorphins”。如对BMP途径激活剂所期望的，它们诱导C2C12成肌细胞分化为成骨细胞。Affymetrix RNA分析证实了分化结果，并表明文氏吗啡处理可引起类似于BMP4处理的遗传反应。与异黄酮异黄酮（SJ000286237）不同，黄酮在治疗24小时后能最大程度地激活该途径，而所有三种ventromorphins均在治疗30分钟内诱导SMAD1 / 5/8磷酸化，并在1 h内达到峰值活性，这表明它们的反应与直接激活BMP信令。