Matthes E, von Janta-Lipinski M, Will H, Schröder H C, Merz H, Steffen R, Müller W E
Institut für Molekularbiologie, Berlin-Buch, Germany.
Biochem Pharmacol. 1992 Apr 1;43(7):1571-7. doi: 10.1016/0006-2952(92)90216-6.
The effect of analogues of both 2',3'-dideoxy-3'-fluorothymidine (FddThd) [2',3'-dideoxy-3'-fluorouridine (FddUrd), 2',3'-dideoxy-3'-fluoro-5-chlorouridine (FddClUrd), 2',3'-dideoxy-3'- fluoro-5-bromouridine (FddBrUrd) and 2',3'-dideoxy-3'-fluoro-5-bromovinyluridine (FddBVUrd)] and 2',3'-dideoxy-3'-fluorocytidine (FddCyt) [2',3'-dideoxy-3'-fluoro-5-fluorocytidine (FddFCyt), 2',3'-dideoxy-3'-fluoro-5-chlorocytidine (FddClCyt), 2',3'-dideoxy-3'-fluoro-5-methylcytidine (FddMeCyt), 2',3'-dideoxy-3'-fluoro-5-ethylcytidine (FddEtCyt), 2',3'-dideoxy-3'-chloro-5-methylcytidine (ClddMeCyt), 2',3'-dideoxy-3'-amino-5-methylcytidine (AmddMeCyt), 2',3'-dideoxy-3'-azido-5- methylcytidine (AzddMeCyt) and arabinosyl-5-methylcytosine (AraMeCyt)] were tested for their potential antiviral activity in vitro using the human hepatoblastoma cell line, Hep G2 2.2.15, which was transfected with a vector containing hepatitis B virus (HBV). It was found that FddThd, FddMeCyt, FddEtCyt, ClddMeCyt, AmddMeCyt and AraMeCyt display cytostatic activity at concentrations (CD50 values) between 0.54 (FddMeCyt) and 3.93 microM (FddEtCyt), while FddUrd, FddClUrd, FddBrUrd, FddBVUrd, FddCyt, FddFCyt, FddClCyt and AzddMeCyt do not affect cell growth at concentrations of up to 25 microM. Among the thymidine analogues tested, FddThd is the most effective antiviral agent: at a concentration of 0.03 microM a more than 90% reduction of HBV DNA synthesis was measured. On the other hand, the antiviral indexes displayed by FddClUrd, FddBrUrd and FddBVUrd are higher than tht of FddThd; FddUrd was completely inactive. The most powerful antiviral agents in the group of cytidine analogues tested in vitro were FddMeCyt (more than 90% reduction of HBVDNA synthesis at 0.10 microM) and ClddMeCyt (0.10 microM); FddClCyt, FddEtCyt, AmddMeCyt and AraMeCyt were of intermediate activity. None of the negligible antiviral activity was determined for FddUrd, FddCyt, FddFCyt and AzddMeCyt. FddThd and FddMeCyt displayed in vivo an antiviral effect in the duck/duck HBV (DHBV) animal system. Administration of 10 or 20 mg/kg (total daily dose) of FddThd and 5 or 10 mg/kg of FddMeCyt (i.m. daily) to ducks infected with DHBV for 12 days blocked virus production. Termination of treatment with FddThd of infected animals led to reappearance of the virus in the serum though at lower levels. The in vitro and the in vivo data suggest that FddThd and FddMeCyt might be promising antiviral agents for the treatment of infection caused by HBV in humans.
使用转染了含乙型肝炎病毒（HBV）载体的人肝癌细胞系Hep G2 2.2.15，在体外测试了2',3'-二脱氧-3'-氟胸苷（FddThd）的类似物[2',3'-二脱氧-3'-氟尿苷（FddUrd）、2',3'-二脱氧-3'-氟-5-氯尿苷（FddClUrd）、2',3'-二脱氧-3'-氟-5-溴尿苷（FddBrUrd）和2',3'-二脱氧-3'-氟-5-溴乙烯基尿苷（FddBVUrd）]以及2',3'-二脱氧-3'-氟胞苷（FddCyt）的类似物[2',3'-二脱氧-3'-氟-5-氟胞苷（FddFCyt）、2',3'-二脱氧-3'-氟-5-氯胞苷（FddClCyt）、2',3'-二脱氧-3'-氟-5-甲基胞苷（FddMeCyt）、2',3'-二脱氧-3'-氟-5-乙基胞苷（FddEtCyt）、2',3'-二脱氧-3'-氯-5-甲基胞苷（ClddMeCyt）、2',3'-二脱氧-3'-氨基-5-甲基胞苷（AmddMeCyt）、2',3'-二脱氧-3'-叠氮基-5-甲基胞苷（AzddMeCyt）和阿拉伯糖基-5-甲基胞嘧啶（AraMeCyt）]的潜在抗病毒活性。发现FddThd、FddMeCyt、FddEtCyt、ClddMeCyt、AmddMeCyt和AraMeCyt在浓度（CD50值）介于0.54（FddMeCyt）和3.93微摩尔/升（FddEtCyt）之间时表现出细胞生长抑制活性，而FddUrd、FddClUrd、FddBrUrd、FddBVUrd、FddCyt、FddFCyt、FddClCyt和AzddMeCyt在浓度高达25微摩尔/升时不影响细胞生长。在所测试的胸苷类似物中，FddThd是最有效的抗病毒剂：在浓度为0.03微摩尔/升时，测得HBV DNA合成减少超过90%。另一方面，FddClUrd、FddBrUrd和FddBVUrd显示的抗病毒指数高于FddThd；FddUrd完全无活性。在所测试的胞苷类似物组中，最有效的抗病毒剂是FddMeCyt（在0.10微摩尔/升时HBV DNA合成减少超过90%）和ClddMeCyt（0.10微摩尔/升）；FddClCyt、FddEtCyt、AmddMeCyt和AraMeCyt具有中等活性。未测定FddUrd、FddCyt、FddFCyt和AzddMeCyt的可忽略不计的抗病毒活性。FddThd和FddMeCyt在鸭/鸭HBV（DHBV）动物系统中显示出体内抗病毒作用。对感染DHBV 12天的鸭每天肌肉注射10或20毫克/千克（每日总剂量）的FddThd以及5或10毫克/千克的FddMeCyt可阻断病毒产生。感染动物停止使用FddThd治疗后，病毒在血清中重新出现，不过水平较低。体外和体内数据表明，FddThd和FddMeCyt可能是治疗人类HBV感染的有前景的抗病毒剂。
