WHI-P154 (Jak3 inhibitor ii) is a potent JAK3 inhibitor.

Src:100 nM, EGFR:4 nM, VEGFR:100 nM, JAK3:1.8 μM

体内使用EGF-P154导致严重联合免疫缺陷小鼠成胶质细胞瘤异种移植模型中的肿瘤生长延迟并且改善无瘤存活.然而无瘤小鼠生存期都没有超过33天 (无瘤生存期中值为19天),并且所有对照小鼠的肿瘤在58天内迅速达到平均尺寸> 500 mM3,用1 mg/kg/天 EGF-P154连续治疗10天的小鼠中有40%活下来并且58天以上没有检测到肿瘤,无瘤生存期中值为40天.剩下的60%小鼠中肿瘤体积从未超过50 mm3.

WHI-P154的体外抗胶质母细胞瘤活性扩增> 200倍,并通过结合重组人源表皮生长因子而具有选择性。WHI-P154在体外抑制巨噬细胞中STAT1激活,iNOS表达和NO产生。研究证明,WHI-P154还可以抑制其它常见激酶,包括EGFR,Src,Abl,VEGFR,MAPK和PI3-K,并诱导人成胶质细胞瘤细胞系中的细胞凋亡。WHI-P154在ECM中抑制胶质母细胞瘤细胞粘附和迁移。 WHI-P154 对人恶性胶质瘤细胞系U373和U87具有明显的细胞毒性,在微摩尔浓度下引起凋亡性细胞死亡。

Kinase assays: WHI-P154 is tested in kinase assays. The panel of kinases is selected to broadly cover the kinome, providing a good approximation of specificity. For all kinases, recombinant rat (IKKβ) or human (all others), full-length or GST-kinase domain fusion proteins, are used. WHI-P154 is inactive (concentration that inhibits response by 50% [IC50] > 30 μM) for the following kinases: AKT, AuroraA, cdk2, cdk6, CHK1, FGFR1, GSK3b, IKKb, IKKi, INSR, MAPK1, MAPKAP-K2, MASK, MET, PAK4, PDK1, PKCb, ROCK1, TaoK3, TrkA.

Cells are seeded into a 96-well plate at a density of 2.5×104 cells/well and incubated for 36 h at 37 ℃ before drug exposure. On the day of treatment, culture medium is carefully aspirated from the wells and replaced with fresh medium containing the quinazoline compounds WHI-P154 at concentrations ranging from 0.1 μM to 250 μM. Triplicate wells are used for each treatment. The cells are incubated with the compound for 24hours to 36hours at 37 ℃ in a humidified 5% CO2 atmosphere. To each well, 10 μL of MTT (final concentration, 0.5 mg/mL) is added, and the plate are incubated at 37 ℃ for 4 h. Than solubilized overnight at 37 ℃ in a solution containing 10% SDS in 0.01 M HCL. The absorbance of each well is measured in a microplate reader at 570 nm.(Only for Reference)