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.
出处
《中国药科大学学报》
CAS
CSCD
北大核心
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)~~
