HPLC测定叶酸-青霉素G酰化酶对N-苯乙酰化阿霉素的催化活性  

HPLC Determination of Enzymatic Hydrolysis of N-(Phenylacetyl) Doxorubicin Catalyzed by Folate-conjugated Penicillin-G Acylase

作　　者：张奇[1] 项光亚[1] 龙娜[1] 吴继洲[1] 

机构地区：[1]华中科技大学同济医学院药学院,武汉430030

出　　处：《中国药科大学学报》2005年第6期535-537,共3页Journal of China Pharmaceutical University

基　　金：国家自然科学基金资助项目(No.30300430)~~

摘　　要：目的：利用高效液相色谱法测定叶酸-青霉素G酰化酶（PGA）对N-苯乙酰化阿霉素（DOXP）的催化活性，为叶酸导向的酶催化前体药物的肿瘤治疗（FDEPF）研究奠定理论基础。方法：色谱柱为Diamonsil C18（250mm×4．6mm，5μm）；流动相为乙腈-水，用85％磷酸调节pH至2．4；梯度程序：0.3min为24％乙腈，3～15min为80％乙腈；紫外检测波长：495nm；流速：1mL/min。结果：DOXP和酶解产物阿霉素的tR分别为5．5min和12．3min，原酶PGA和叶酸-PGA偶联酶对DOXP的Km和Vmax分别为15μmol/mL，0．094μmol/（min·mg）和19μmol/mL，0．086μmal/（min·mg）。结论：DOXP为PGA的较好底物，叶酸与PGA偶联对酶的催化活性的影响较小。AIM： To investigate enzymatic hydrolysis of N-（phenylacetyl） doxorubicin （DOXP） catalyzed by folate-conjugated penicillin-G acylase （PGA）. METHODS： The determination of prodrug DOXP and cleavage product doxorubicin was monitored by HPLC.A Diamonsil C18 column （250 mm×4.6 mm, 5μm） was used with 1 mL/min flow rate of 85% phosphoric acid in water （pH 2.4） with a gradient of acetonitrile created according to the following scheme： 24% for 3 rain and then constant at 80% for 12 rain. RESULTS： PGA effected the hydrolysis of DOXP, and HPLC analysis confirmed that doxorubicin was formed. The Km and Vmax were 15μmol/mL, 0.094μmol/（min-mg） and 19μmol/mL, 0.086μmol/（min·mg） for the parent enzyme or for the conjugate enzyme, respectively. CONCLUSION： DOXP was a good substrate for PGA. Folate-conjugated PGA did not significantly compromise PGA catalytic activity.

关 键 词：叶酸 青霉素G酰化酶 N-笨乙酰化阿霉素 催化活性 高效液相色谱法 

分 类 号：R91[医药卫生—药学]