Emergence of resistance to carbocyclic oxetanocin G in herpes simplex virus type 1 and genetic analysis of resistant mutants

作     者：Hiroshi SHIOTA 

作者机构：Department of Ophthalmology Japan School of Medicine The University of Tokushima Tokushima 770 

出 版 物：《Acta Pharmacologica Sinica》 (中国药理学报(英文版))

年 卷 期：2004年第25卷第7期

页      面：75-80页

核心收录：

学科分类：1007[医学-药学(可授医学、理学学位)] 1006[医学-中西医结合] 100706[医学-药理学] 1002[医学-临床医学] 100602[医学-中西医结合临床] 10[医学] 

主　　题：carbocyclic oxetanocin G human herpesvirus 1 viral drug resistance thymidine kinase 

摘      要：AIM: To elucidate the potentiality of emergence of drug-resistance to carbocyclic oxetanocin G (***-G ), a new effective antiviral drug for herpetic keratitis during treatment and the mechanism of this drug resistance. METHODS: A ***-G resistant strain (***-Gr) was established by serially propagating the herpes simplex virus (HSV) -1 in African green monkey kidney (VERO) cells in the presence of ***-G. After the drug sensitiv- ity assay and the thymidine kinase (TK) activity assay, the molecular basis for the drug resistance was studied using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis and PCR direct se- quencing technology. RESULTS: After the 10th passage in 10 μm ***-G, the ED50 of the ***-Gr was 17.08-fold greater than that of the original strain on the average and the TK activities of these resistant strains were extremely reduced. PCR-SSCP analysis on TK gene of the wild HSV-1 and the ***-Gr showed altered migration patterns in part 3 and part 4, while PCR-SSCP analysis on DNA polymerase gene showed no difference among the viruses. Sequence analysis revealed a deletion of G at position of 430 that caused frameshift, resulting in premature termination in the TK gene. CONCLUSION: The drug resistance to ***-G may appear during the treatment due to the deficiency of TK activity caused by a single mutation in the TK gene of HSV-1.