Dasatinib monohydrate 是一种具有口服活性的，ATP 竞争性的双重Src/Bcr-Abl抑制剂，有抗肿瘤活性，还诱导凋亡和自噬。它抑制Src和Bcr-Abl的IC50分别为 0.5 nM 和<1.0 nM，Ki值分别为 16 pM 和 30 pM。

Dasatinib is an orally bioavailable synthetic small molecule-inhibitor of SRC-family protein-tyrosine kinases. Dasatinib binds to and inhibits the growth-promoting activities of these kinases. Apparently, because of its less stringent binding affinity for the BCR-ABL kinase, dasatinib has been shown to overcome the resistance to imatinib of chronic myeloid leukemia (CML) cells harboring BCR-ABL kinase domain point mutations.

在TgE小鼠体内,Dasatinib特异性抑制骨髓B细胞（LMP2A表达）形成集落,并使脾脏大小减少.与对照组Tg6/λ-MYC小鼠相比,Dasatinib处理组脾脏肿块显著变小.Dasatinib对MYC/LMP2A双重转基因小鼠体内的淋巴结肿大具有抑制作用,并可逆转脾肿大.从LMP2A/MYC双重转基因小鼠移植肿瘤细胞至Rag1KO小鼠,Dasatinib可逆转该脾肿大.Dasatinib可抑制B淋巴肿瘤细胞（表达LMP2A）中Lyn磷酸化.

Dasatinib对野生型Abl激酶及其所有突变体（除T315I外）具有有效的抑制作用。Dasatinib直接靶向所有突变型Abl激酶域,浓度依赖性地抑制底物和自身磷酸化。与伊马替尼相比,Dasatinib对表达Bcr-Abl（野生型和除T315I外的突变型）的Ba/F3细胞增殖具有更有效的抑制作用。与伊马替尼相比,Dasatinib对表达野生型Bcr-Abl的细胞表现出325倍的效能。在Dasatinib处理后TgE骨髓细胞集落百分数从100%降为4.12%。在Dasatinib存在下,WT和TgE骨髓细胞形成的集落百分比具有明显的统计学差异。Dasatinib抑制LMP2A表达引起的B淋巴细胞存活和增殖。 Dasatinib可抑制部分甲状腺癌细胞的Src信号,抑制细胞生长,并诱导细胞凋亡和细胞周期阻滞。以Dasatinib（0.019 μM~1.25 μM）治疗3天,抑制大约50%的BCPAP,C643,SW1736和TPC1细胞系生长,但需要更高的浓度才能抑制K1细胞系生长。在BCPAP,SW1736和K1细胞中,Dasatinib（10 nM或50 nM）可使G1期细胞增加9-22%,而S期细胞相应减少7-18%。

Kinase autophosphorylation assays: Kinase assays using wild-type and mutant glutathione S-transferase (GST)-Abl fusion proteins (c-Abl amino acids 220-498) are done. GST-Abl fusion proteins are released from glutathione-Sepharose beads before use; the concentration of ATP is 5 μM. Immediately before use in kinase autophosphorylation and in vitro peptide substrate phosphorylation assays, GST-Abl kinase domain fusion proteins are treated with LAR tyrosine phosphatase. After 1-hour incubation at 30 °C, LAR phosphatase is inactivated by addition of sodium vanadate (1 mM). Immunoblot analysis comparing untreated GST-Abl kinase to dephosphorylated GST-Abl kinase is routinely done using phosphotyrosine-specific antibody 4 g10 to confirm complete (>95%) dephosphorylation of tyrosine residues and c-Abl antibody CST 2862 to confirm equal loading of GST-Abl kinase. The Dasatinib concentration range is extended to 1,000 nM for mutant T315I. These same inhibitor concentrations are used for the in vitro peptide substrate phosphorylation assays. The three inhibitors are tested over these same concentration ranges against GST-Src kinase and GST-Lyn kinase.

Ba/F3 cell lines are seeded in triplicate and incubated with escalating concentrations of Dasatinib for 72 hours. Proliferation is measured using a methanethiosulfonate-based viability assay. IC50 and IC90 values are reported as the mean of three independent experiments done in quadruplicate. The inhibitor concentration ranges are 0 nM to 32 nM (Dasatinib). The Dasatinib concentration range is extended to 200 nM for mutant T315I.(Only for Reference)

863127-77-9

C22H28ClN7O3S

506.02

BMS-354825 Monohydrate;达沙替尼

Ethanol:<1 mgml
H2O:<1 mgml
DMSO:20 mg/mL (39.5 mM)
Powder: -20°C for 3 years
In solvent: -80°C for 2 years