摘要
TM-2 known as a potential antitumor drug is a novel semi-synthetic taxane derivative. As drug-protein interactions contribute to insights into pharmacokinetic and pharmacodynamic properties, we eluci- dated the binding of TM-2 to plasma protein. In this study, a simple, rapid and reliable method was developed and validated employing equilibrium dialysis for the separation of bound and unbound drugs and ultra-performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS） for the quantitation. Protein binding reached equilibrium within 24 h of incubation at 37 ℃. After liquid-liquid extraction with methyl tert-butyl ether, the samples were separated on Thermo Syncronis UPLC C18 （2.1 mm× 50 mm, 1.7 μm）, and acquisition of mass spectrometric data was performed in multiple re- action monitoring （MRM） mode via positive electrospray ionization. The assay was linear over the concentration rang of 5-2000 nglmL The intra- and inter-day precisions were 0.1%-14.8%, and the accuracy was from -6.4% to Z0%. This assay has been successfully applied to a protein binding study of TM-2 in rat, human and beagle dog plasma. TM-2 showed high protein binding of 81.4% ± 6.5% （rat）, 87.9% ± 3.6% （human） and 79.4% ± 4.0% （beagle dog）. The results revealed that there was an insignificant difference among the three species.
TM-2 known as a potential antitumor drug is a novel semi-synthetic taxane derivative. As drug-protein interactions contribute to insights into pharmacokinetic and pharmacodynamic properties, we eluci- dated the binding of TM-2 to plasma protein. In this study, a simple, rapid and reliable method was developed and validated employing equilibrium dialysis for the separation of bound and unbound drugs and ultra-performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS） for the quantitation. Protein binding reached equilibrium within 24 h of incubation at 37 ℃. After liquid-liquid extraction with methyl tert-butyl ether, the samples were separated on Thermo Syncronis UPLC C18 （2.1 mm× 50 mm, 1.7 μm）, and acquisition of mass spectrometric data was performed in multiple re- action monitoring （MRM） mode via positive electrospray ionization. The assay was linear over the concentration rang of 5-2000 nglmL The intra- and inter-day precisions were 0.1%-14.8%, and the accuracy was from -6.4% to Z0%. This assay has been successfully applied to a protein binding study of TM-2 in rat, human and beagle dog plasma. TM-2 showed high protein binding of 81.4% ± 6.5% （rat）, 87.9% ± 3.6% （human） and 79.4% ± 4.0% （beagle dog）. The results revealed that there was an insignificant difference among the three species.
基金
partly supported by the National High Technology Research and Development Program of China(No.2012AA020305)
