重组人血白蛋白与小分子药物结合功能研究.pdfVIP

2006 9 19 5 Chin J Biologicals September 2006, Vo1. 19 No. 5 499 陶苒李梅彦王慧欣 董爱华贾茜 366 nm , ; , SephadexG-25HPLC, , 1468 molD iaze- pam /mol rHS , 1282 molD iazepam /molHS 869%, 673% ; ; Binding Function ofRecombinantHuman Serum Albumin to SmallMoleculeDrugs T O Ran, LIMe-iyan,W NG Hui-xin, et al D evelopm ent Company of N ew D rugs, N orth Ch ina Pharmaceutical In- dustry Group, Sh ij iazhuang 050015, China Abstract Objective To study the binding function of recombinant human serum album in HS to smallmolecule drugs. M ethods Determ ine the 366 values ofD iazepam before and after binding toHS and calculate the binding constant. Bind warfarin to HS , and separate the conjugate and freewarfarin by Sephadex G-25HPLC, then integrate the peak areas of conjugate and freewarfarin to calculate the binding rate. Results The binding constantof recombinantHS to Diazepam was 1468molDiazepam /mol rHS , and that ofHS to Diazepam was 1282molDiazepam /molHS . The binding rates ofwarfarin in rHS andHS were 869% and 673% respectively. Conclusion The binding functions of recombinant and natureHS to smallmolecule drugswere basically in agreement. Key ords Human serum albumin; Smallmolecule drug; Binding function Hum an serum album in, HS , , 585, [ 1, 2] , 66 500,, , 4235 g/L , , , HS 1 Spectronic Genesys, SPECTRON- IC INSTRUMENT, INC ; pHs-3C , , ; Waters 515 HPLC Pump W aters ; Waters 996 Photodiode rray Detector W aters HS , 2 Recombinant hu- Diazepam Warfarin , man serum album in, rHS Sigma; rHS 200 mg/m l , ; HS 200 mg/m l , : 863 2002 2Z345C . ; Sephadex G-25 desalting XK16/20, Vt : 10m l , 050015 . 3 : , E-mail: taoran197806@ yahoo. com. cn 180 2 h 500 2006 9 19 5 Chin J Biologicals September2006, Vo1. 19 No. 5 , 2 g HS /g, , , HCl pH 30, 1 h, 2, 366 nm , 15 000 r/m in 20m in,NaOH , [3] pH 70 , , 1, 4 Sephadex G-25 28127 10% 10% 66 500 , HS , 366 nm 1282 molDiazepam/molHS , rHS , - 1468 molDiazepam/mol rHS , 366 nm , 5 67mmol/L pH 74, 863 m