LY-364947 (HTS466284) is a potent ATP-competitive inhibitor of TGFβR-I.

TGFβRII:0.4 μM, RIPK2:0.11 μM, CK1δ:0.22 μM, TGFβRI:59 nM, MLK7K:1.4 μM

1 mg/kg i.p. LY364947在BxPC3胰腺癌细胞的肿瘤异种移植模型中,明显增加肿瘤组织中LYVE-1-阳性区域.25 mg/kg i.p. LY364947在慢性腹膜炎小鼠模型中,明显增加的LYVE-1-阳性区域表明其加速淋巴管生成.Y364947（25 mg/kg）可增加CML感染小鼠中白血病起始细胞中的p-Akt和减少核Foxo3a.

在浓度低至0.25 μM时,LY364947使NMuMg细胞中的xVent2-lux BMP4响应增强30％。2 μM LY364947防止NMuMg细胞中TGF-β诱导的上皮 - 间质转化。3 μM LY364947处理24小时后,几乎在所有HDLECs中诱导Prox1和LYVE-1表达。LY364947促进Foxo3a的核输出,在白血病阳性细胞中具有低Smad2/3和高Akt磷酸化水平。 与OP-9基质细胞共培养后,LY364947（<20 μM）抑制白血病起始细胞集落形成能力。LY364947作为一种ATP竞争性,紧密结合抑制剂,通过TGFβR-I激酶抑制P-Smad3磷酸化,Ki为28 nM。LY364947抑制体内Smad2磷酸化,NMuMg细胞中IC50为135 nM。LY364947抑制NMuMg细胞内的体内Smad2磷酸化,IC50为135 nM。

The IC50 of LY-364947 at different enzyme concentrations are determined by the filter-binding assay. Typically, 40 μL reactions in 50 mM HEPES at pH 7.5, 1 mM NaF, 200 μM pKSmad3(-3), and 50 mM ATP containing a titration of each inhibitor with concentrations of 1600, 800, 400, 200, 100, 50, 25, and 0 nM are incubated at 30°C for 30 min. The IC50 is calculated using a nonlinear regression method with GraphPad Prism software. The binding type is determined by plotting the correlation between enzyme concentrations and IC50 values.