Sunitinib 是一种 ATP 竞争性抑制剂，通过抑制自身磷酸化和 RNase 激活来有效抑制IRE1α的磷酸化。它还是一种多靶点受体酪氨酸激酶抑制剂，抑制VEGFR2和PDGFRβ的IC50分别为 80 nM 和 2 nM

Sunitinib, a multi-targeted RTK inhibitor, is targeting PDGFRβ and VEGFR2 (Flk-1) with IC50 of 2 nM and 80 nM and also inhibits c-Kit.

在FLT3-ITD骨髓移植模型中,Sunitinib（每天20 mg/kg）对皮下MV4;11 (FLT3-ITD)异种移植物的生长具有明显抑制作用,并延长生存.与实质性,选择性抑制VEGFR2或PDGFR在体内的磷酸化和信号传导一致,Sunitinib（每天20-80 mg/kg）对各种肿瘤异种移植模型,包括HT-29,Colo205,A431,SF763T,C6,H-460,A375或MDA-MB-435表现出广泛有效的剂量依赖性抗肿瘤活性.Sunitinib（每天80 mg/kg）给药21天,8只小鼠中有6只的肿瘤完全消退,且在治疗结束后的110天的观察期内肿瘤没有再生.尽管不能完全恢复到第一轮治疗的情况,但在第二轮使用Sunitinib治疗仍然能够有效抗肿瘤.Sunitinib治疗可使肿瘤MVD明显下降,如在SF763T神经胶质瘤中可减少~40%.尽管肿瘤没有减小,SU11248治疗还是能够完全抑制表达荧光素酶的PC-3M异种移植的肿瘤生长.

Sunitinib是PDGFRβ（Ki：8 nM）和 VEGFR2 (Flk1)（Ki：9 nM）有效的ATP竞争性抑制剂。与FGFR-1,EGFR,Cdk2,Met,IGFR-1,Abl,和src相比,Sunitinib作用于VEGFR2和PDGFR的选择性要高出10倍。在血清饥饿的表达PDGFRβ或VEGFR2的NIH-3T3细胞中,Sunitinib抑制VEGF依赖性VEGFR2磷酸化和PDGF依赖性PDGFRβ磷酸化,IC50均为10 nM。Sunitinib抑制野生型FLT3-ITD,FLT3-Asp835和FLT3磷酸化,IC50分别为50 nM,30 nM和250 nM。Sunitinib抑制OC1-AML5和MV4;11 细胞的增殖,IC50分别为14 nM和8 nM,并以剂量依赖的方式诱导细胞凋亡。Sunitinib能够有效抑制FLT-3和Kit。对于过表达PDGFRα或PDGFRβ的NIH-3T3细胞,Sunitinib抑制VEGF对其诱导的增殖,IC50 分别为69 nM和39 nM。

Biochemical Tyrosine Kinase Assays: IC50 values for Sunitinib against VEGFR2 (Flk-1) and PDGFRβ are determined using glutathione S-transferase fusion proteins containing the complete cytoplasmic domain of the RTK. Biochemical tyrosine kinase assays to quantitate the trans-phosphorylation activity of VEGFR2 (Flk-1) and PDGFRβ are performed in 96-well microtiter plates precoated (20 μg/well in PBS; incubated overnight at 4 °C) with the peptide substrate poly-Glu,Tyr (4:1). Excess protein binding sites are blocked with the addition of 1-5% (w/v) BSA in PBS. Purified GST-fusion proteins are produced in baculovirus-infected insect cells. GST-VEGFR2 and GST-PDGFRβ are then added to the microtiter wells in 2 × concentration kinase dilution buffer consisting of 100 mM HEPES, 50 mM NaCl, 40 μM NaVO4, and 0.02% (w/v) BSA. The final enzyme concentration for GST-VEGFR2 or GST-PDGFRβ is 50 ng/mL. Twenty-five μL of diluted Sunitinib are subsequently added to each reaction well to produce a range of inhibitor concentrations appropriate for each enzyme. The kinase reaction is initiated by the addition of different concentrations of ATP in a solution of MnCl2 so that the final ATP concentrations spanned the Km for the enzyme, and the final concentration of MnCl2 is 10 mM. The plates are incubated for 5-15 minutes at room temperature before stopping the reaction with the addition of EDTA. The plates are then washed three times with TBST. Rabbit polyclonal antiphosphotyrosine antisera are added to the wells at a 1:10,000 dilution in TBST containing 0.5% (w/v) BSA, 0.025% (w/v) nonfat dry milk, and 100 μM NaVO4 and incubated for 1 hour at 37 °C. The plates are then washed three times with TBST, followed by the addition of goat antirabbit antisera conjugated with horseradish peroxidase (1:10,000 dilution in TBST). The plates are incubated for 1 hour at 37 °C and then washed three times with TBST.The amount of phosphotyrosine in each well is quantitated after the addition of 2,2′-azino-di-[3-ethylbenzthiazoline sulfonate] as substrate.

Cells are starved overnight in medium containing 0.1% FBS prior to addition of Sunitinib and FL (50 ng/mL; FLT3-WT cells only). Proliferation is measured after 48 hours of culture using the Alamar Blue assay or trypan blue cell viability assays. Apoptosis is measured 24 hours after Sunitinib addition by Western blotting to detect cleavage of poly (ADP-ribose) polymerase (PARP) or levels of caspase-3. (Only for Reference)

557795-19-4

C22H27FN4O2

398.482

苏尼替尼;舒尼替尼;SU 11248

H2O:<1 mgml
DMSO:24 mg/mL (60.2 mM)
Ethanol:<1 mgml
Powder: -20°C for 3 years
In solvent: -80°C for 2 years