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

Dasatinib is a potent inhibitor of the Bcr-Abl and Src family (IC50s: 0.6, 0.8, 79 and 37 nM for Abl, Src, c-Kit, and c-KitD816V, respectively).

Dasatinib (BMS-354825) potently inhibited wild-type Abl kinase and all mutants except T315I over a narrow range (IC50 ≤ 1.7 nmol/L). BMS-354825 (IC50: 0.8 nmol/L) displayed 325-fold greater potency compared with imatinib against cells expressing wild-type Bcr-Abl [1]. BMS-354825 binds 76 of 148 kinases screened at 10 μM, 47 of them with Kd 200 nM [2]. Dasatinib inhibits the kinase activity of KITD816V with comparable efficiency to wild-type KIT (IC50 of 37 and 79 nM, respectively) in in vitro kinase experiments. Dasatinib suppresses the growth of HMC-1.1V560G+, D816V- cells in the low nanomolar range. Furthermore, dasatinib retains activity against HMC-1.2V560G+, D816V+ cells [3].

Daily treatment with dasatinib (50 mg/kg) was initiated on day 10. Using this approach, a significant inhibition of BCPAP orthotopic tumor growth was observed 6 days after treatment, which was sustained through days 23 and 29, compared with vehicle-treated mice. The BCPAP orthotopic final tumor volumes were inhibited by more than 90% in response to dasatinib treatment [4]. The in vivo PK values in rat plasma and DBS after oral administration (50 mg/kg dasatinib) were comparable. The mean AUC value at 10 mg/kg from the historical report was about1200 ng·h/mL while themeanAUCvalue at 50 mg/kg in rat plasma from this study was about 2000 ng·h/mL [5].

Kinase assays using wild-type and mutant glutathione S-transferase (GST)–Abl fusion proteins (c-Abl amino acids 220-498) were done as described, with minor alterations. GST-Abl fusion proteins were released from glutathione-Sepharose beads before use; the concentration of ATP was 5 μmol/L. Immediately before use in kinase autophosphorylation and in vitro peptide substrate phosphorylation assays, GST-Abl kinase domain fusion proteins were treated with LAR tyrosine phosphatase according to the manufacturer's instructions. After 1-hour incubation at 30°C, LAR phosphatase was inactivated by addition of sodium vanadate (1 mmol/L). Immunoblot analysis comparing untreated GST-Abl kinase to dephosphorylated GST-Abl kinase was routinely done using phosphotyrosine-specific antibody 4G10 to confirm complete (>95%) dephosphorylation of tyrosine residues and c-Abl antibody CST 2862 to confirm equal loading of GST-Abl kinase. The inhibitor concentration ranges for IC50 determinations were 0 to 5,000 nmol/L (imatinib and AMN107) or 0 to 32 nmol/L (BMS-354825). The BMS-354825 concentration range was extended to 1,000 nmol/L for mutant T315I. These same inhibitor concentrations were used for the in vitro peptide substrate phosphorylation assays. The three inhibitors were tested over these same concentration ranges against GST-Src kinase and GST-Lyn kinase [1].

Ba/F3 cell lines were plated in triplicate and incubated with escalating concentrations of imatinib, AMN107, or BMS-354825 for 72 hours. Proliferation was 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 for IC50 and IC90 determinations were 0 to 2,000 nmol/L (imatinib and AMN107) or 0 to 32 nmol/L (BMS-354825). The imatinib concentration range was extended to 6,400 nmol/L for mutants with IC50 >2,000 nmol/L. The BMS-354825 concentration range was extended to 200 nmol/L for mutant T315I [1].

For in vivo studies, dasatinib (50 mg/kg) was prepared for daily oral gavage (5 d/wk) in 80 mmol/L sodium citrate buffer, pH 3.0. For the orthotopic murine model, mice were randomized on day 10 based on bioluminescence activity to receive drug or vehicle. In the metastatic murine model, mice received dasatinib or vehicle, as described earlier, starting 2 days before intracardiac injection (pretreatment), or on day 11 following randomization (posttreatment) [4].

302962-49-8

C22H26ClN7O2S

488.01

BMS-354825;达沙替尼

DMSO:91 mg/mL (186.5 mM)
Ethanol:<1 mgml
H2O:<1 mgml
Powder: -20°C for 3 years
In solvent: -80°C for 2 years