重组人血清白蛋白/干扰素α2a融合蛋白的药效学、药代动力学及安全性评价  

Pharmacodynamics, pharmacokinetics and safety studies on recombinant human serum albumin/interferon-α2a fusion protein

作　　者：富岩 马金玲 孙丽 杜杰 张延林 徐楠楠 孙云霞 于在林 

机构地区：[1]北京美福源生物医药科技有限公司,100085 [2]天津林达生物科技有限公司,300457 [3]天津溥瀛生物技术有限公司,300457 [4]北京昭衍新药研究中心有限公司,100176

出　　处：《中国医药生物技术》2012年第6期401-411,共11页Chinese Medicinal Biotechnology

基　　金：北京市海淀区科学技术委员会(K20100133Z);国家"重大新药创制"专项(2009ZX09301-008N20);天津市科学技术委员会(07ZCKFSH02400);天津科技型中小企业技术创新基金(11ZXCXST05000)

摘　　要：目的对具有独立自主知识产权的注射用重组人血清白蛋白／干扰素α2a融合蛋白新药的临床前药效学、药代动力学和安全性评价进行研究。方法通过重组人血清白蛋A／干扰素α2a融合蛋白（rHS-IFNα2a）对HepG22．2．15分泌乙型肝炎病毒（HBV）细胞作用产生的抗病毒进行体外药效学评价；以给药后的食蟹猴体内2＇-5’寡腺昔酸合成酶活性作为干扰素药效学标志基因表达考察药效动力学指标；药代动力学观察了不同剂量的挖^125rHSA／IFNα2a单次皮下注射给予小鼠、大鼠后，各组织器官的分布、排泄情况及rHSA／IFNα2a单次和多次皮下注射给予食蟹猴后其体内的代谢情况：安全性评价中，安全药理学研究观察了rHSA／IFNα2a对小鼠中枢神经系统及对狗呼吸和心血管系统功能的影响：毒理学研究观察了rHSA／IFNα2a单次注射给予小鼠（皮下和静脉）、食蟹猴（皮下）的毒性反应，及反复皮下注射给予大鼠和食蟹猴的毒性反应，食蟹猴反复给药的毒性试验中还伴行了毒代动力学的研究。妊娠Wistar大鼠皮下注射给予rHSA／IFNα2a对孕鼠生殖能力及胎仔的影响。结果试验药物rHSA／IFNα2a和阳性对照品（PEG-rhlFNα2a）的最大无毒剂量为3．13×10^3IU／ml，当以低于最大无毒剂量的rHSA／IFNα2a和阳性对照品处理HepG22．2．15细胞，显示细胞分泌HBV病毒的能力受到抑制。细胞上清液中的HBV核酸量随时间而减少，HBV的S抗原和e抗原表达也同步受到抑制，与PEG．rhlFNα2a对HBV病毒分泌的相对抑制率检测结果相同。药代动力学研究结果显示静脉rHSA／IFNα2a给药组和对照组的t1／2分别为（88．88±9．65）h和（35．81±4．91）h：食蟹猴皮下注射末端半衰期较长为61．20-83．57h。安全药理学研究表明单次皮下注射重组人血清白蛋白／干扰素α2a融合蛋白在100、300、600gg／kg剂量范围内对小鼠中枢神�Objective The evaluation of pharmacodynamics, pharmacokinetics and safety in animals of recombinant human serumalbumin/interferon-α2a fusion protein （rHSA/IFNα2a） with independent intellectual property rights. Methods For pharmacodynamics study, in vitro antiviral effect of rHSA/IFNα2a on inhibiting the secretion of hepatitis B virus （HBV） from HepG2 2.2.15 cells was evaluated. Our study also examined the bioactivity of interferon-specific pharmacodynamic marker enzyme （2-5oas） after injection of rHSA/IFNα2a in cynomolgus monkeys. For pharmacokinetics study, the distribution and excretion of 125I-rHSA/IFNα2a in different tissues and organs was examined by single subcutaneous injection with different doses in mice and rats. Metabolism in vivo was further performed in cynomolgus monkeys with single and multiple doses. For safety study, effects of rHSA/IFNα2a on the central nervous system in mice as well as the respiratory and cardiovascular system in Beagle dogs were observed. Through acute and chronic toxicity study, the safety evaluation of rHSA/IFNα2a was performed in mice, rats and cynomolgus monkeys with single and repeated doses. The pharmacokinetics/toxicokinetics study was performed to examine the PK parameters. The reproductive toxicity ofrHSA/IFNα2a was also evaluated in pregnant Wistar rats and fetuses. Results The maximum non-toxic dose for rHSA/IFNα2a and the positive control, PEG-rhIFNα2a （Pegasys）, was 3.13×10^3 IU/ml. The secretion of HBV particles was inhibited when the HepG2 2.2.15 cells were treated with a dose lower than the maximum non-toxic amount of rHSA/IFNα2a, and the level of HBV nucleic acid was reduced through the different time periods tested. The expression of HBsAg and HBeAg from HepG2 2.2.15 cells were also inhibited. All the results in vitro were consistent with the relative suppression rate of PEG-rhIFNα2a. Pharmacokinetic study in cynomolgus monkeys through intravenous administration showed the t1/2 was （88.88 ± 9.65） h and （35.81 ± 4.91） h i

关 键 词：白蛋白类 干扰素Α 重组蛋白质类 药代动力学 毒理学 

分 类 号：R96[医药卫生—药理学]