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mg\u002Fkg(几乎完全抑制血栓形成的剂量)下,利伐沙班适度延长PT(3.2±0.5倍)并抑制FXa活性(65±3％)[2]。",{"key":249,"value":250},"激酶实验","利伐沙班（BAY 59-7939）对纯化的丝氨酸蛋白酶的活性使用生色或荧光底物在96孔微量滴定板中于25℃测量。将酶与利伐沙班或其溶剂DMSO一起温育10分钟。通过添加底物引发反应，并且使用Spectra Rainbow Thermo Reader在405nm连续监测颜色或荧光，或者分别使用SPECTRAfluor plus在630 \u002F 465nm监测颜色或荧光20分钟。在以下缓冲液（最终浓度）中分析酶活性：在pH8.3的50mM Tris-HCl缓冲液中的人FXa（0.5nM），兔FXa（2nM），大鼠FXa（10nM）或尿激酶（4nM）。 ，150mM NaCl和0.1％牛血清白蛋白（BSA）;在0.1μMTris-HCl，pH 8.0和20mM CaCl 2中的凝血酶（0.69nM），胰蛋白酶（2.2nM）或纤溶酶（3.2nM）的Pefachrome FXa（50-800μM）或染色体U（250μM）;染色酶TH（200μM），染色质纤溶酶（500μM）或染色酶抑制剂（500μM），FXIa（1nM）或APC（10nM）在50mM磷酸盐缓冲液，pH7.4,150mM NaCl中;和S 2366（150或500μM）FVIIa（1 nM）和组织因子（3 nM）在50 mM Tris-HCl缓冲液，pH 8.0,100 mM NaCl，5 mM CaCl2和0.3％BSA，HD-Phe-Pro中的表达-Arg-6-氨基-1-萘 - 苄基磺酰胺H2O（100μM）并测量3小时。 FIXab \u002F FX测定，包括FIXab（8.8 nM）和FX（9.5 nM）在50 mM Tris-HCl缓冲液，pH 7.4,100 mM NaCl，5 mM CaCl2和0.1％BSA中，通过添加I-1100开始（50μM），并测量60分钟。根据Cheng-Prusoff方程（Ki = IC50 \u002F 1 + [S] \u002F Km）计算针对FXa的抑制常数（Ki），其中[S]是底物浓度，并且Km是Michaelis-Menten常数。 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