JK184 is a potent Hedgehog (Hh) pathway inhibitor.

Hedgehog:30 nM

JK184旨在通过剂量依赖性地抑制胶质瘤（Gli）依赖的转录活动，从而拮抗Hh信号。三天孵化后，JK184显著抑制HUVECs的增殖，IC50为6.3 μg/mL。通过在Panc-1和BxPC-3细胞中使用指定浓度的化合物进行MTT试验，测定JK184的半最大抑制浓度（IC50）（在anc-1中为23.7 ng/mL，在BxPC-3中为34.3 ng/mL），以评估JK184的抗肿瘤效果[1]。与MCF10a、MTSV1-7、HMLE-shGFP和HMLE-pBP细胞相比，Claudin-low细胞系对JK184的治疗更敏感，且JK184在这些细胞中引起的胶质瘤相关的致癌基因同源物1（GLI1）转录和蛋白水平的剂量依赖性减少。使用JK184的IC50剂量治疗可以增加HMLE-shEcad细胞染色与Annexin-V阳性但对碘化丙啶（PI）阴性的比例（P<0.0001，t检验）[2]。

JK184（5 mg/kg，静脉注射）在皮下Panc-1和BxPC-3肿瘤模型中展现了良好的抗增殖活性，并且是针对Hh信号的抗肿瘤化合物的良好候选。组织学分析显示，JK184通过诱导更多的细胞凋亡、降低微血管密度以及减少肿瘤组织中CD31、Ki67和VEGF的表达来增强抗肿瘤活性。然而，JK184的药代动力学特性和生物利用度较差[1]。

JK184 is dissolved in DMSO and stored, and then diluted with appropriate medium before use[1]. The Shh-LIGHT2 cells are seeded in 96-well plates and grown to confluency. The Shh-LIGHT2 cells are treated with various concentrations of JK184 micelles or free JK184 or micelles in DMEM containing 0.5% CS, 0.1 mg/mL streptomycin, 100 U/mL penicillin, 5% Shh-N conditioned medium obtained from Shh-N-producing HEK293 cells. The treated cells are cultured further for 60 h, and firefly and Renilla luciferase activities are measured using a dual luciferase kit. Proliferation assay or apoptosis evaluation of HUVECs is measured using MTT method or FCM analysis, respectively. HUVECs are treated with a series concentration of free JK184, JK184 micelles, or blank MPEG-PCL micelles for 48 h, respectively. The mean percentage of cell inhibition or apoptosis is calculated[1].