Exatecan Mesylate Inhibitor

Solid. Powder

97.32%

Store at -20 degree C for 3 years powder
-80 degree C for 2 years in solvent

531.55

DMSO: 8mg/mL (15.05mM) Need ultrasonic
Water: 6mg/mL (11.29mM) Need ultrasonic and warming
(<1mg/ml refers to the product slightly soluble or insoluble)

C25H26FN3O7S

CS(O)(=O)=O.CC[C@@]1(O)C(=O)OCc2c1cc1-c3nc4cc(F)c(C)c5CC[C@H](N)c(c3Cn1c2=O)c45

169869-90-3

Growth inhibition experiments are carried out in 96-well flat-bottomed microplates, and the amount of viable cells at the end of the incubation is determined by MTT assay. Thus, 500-20000cells/well in 150uL of medium are plated and grown for 24 hours (P388, CCRF-CEM and K562 cells for 4 hours), the drug (including Exatecan Mesylate, in 150uL medium/well), or the medium alone as a control, is added, and the cells are cultured for an additional 3 days. After the addition of MTT (20uL/well, 5mg/mL in phosphate-buffered saline), the plates are incubated for 4 hours and centrifuged at 800g for 5 minutes, then the medium is removed and the blue dye formed is dissolved in 150uL of DMSO. the absorbance is measured at 540nM using a Microplate Reader [1].

Cells (5x10^6) are lysed with SDS buffer (10mM HEPES, 2mM orthovanadate, 10mM NaF, 10mM pyrophosphate, 1mM PMSF, 10ug/mL leupeptin, 10% 2-mercaptoethanol, 10% glycerol, 8% SDS, 42mM Tris-HCl, 0.002% bromophenol blue, pH 7.4). Protein in the whole-cell lysates is separated in 7.5% polyacrylamide gel and blotted onto the nitrocellulose membrane. The membrane is treated with anti-Topo I human antibody and subsequently, with horseradish peroxidase-conjugated protein A. The Topo I-specific band is detected with ECL reagents. To obtain a nuclear extract, cells (5x10^7) are washed with ice-cold buffer (2mM K2HPO4, 5mM MgCl2, 150mM NaCl, 1mM EGTA, 0.1mM dithiothreitol), resuspended in buffer containing 0.35% Triton-X100 and PMSF and then incubated on ice for 10 minutes. The resulting lysates are centrifuged, and precipitates are then incubated with buffer containing 0.35M NaCl for 1 hr at 4 degree C. After centrifugation (18,000g, 10 min), the protein concentration of the supernatant (nuclear extract) is determined using a protein assay kit. The same amount of nuclear protein is analyzed by Western blotting analysis using the anti-Topo I antibody [2].

Exatecan Mesylate (3.325-50mg/kg, i.v.) exhibits antitumor activities in the mice model bearing tumor cells, without toxic death[1]. Exatecan Mesylate (15, 25mg/kg, i.v.) highly inhibits MIA-PaCa, BxPC-3 primary tumor growth in the MIA-PaCa-2 early-stage model and early-stage model of BxPC-3. Exatecan Mesylate (15, 25mg/kg, i.v.) also significantly suppresses BxPC-3 lymphatic metastasis and completely eliminates lung metastasis in the BxPC-3 late-stage cancer model [3].

DNA Damage/DNA Repair; Topoisomerase