靶向HCV IRES和NS3蛋白抑制剂筛选及评价小鼠模型的建立
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摘要
丙型病毒性肝炎是一种严重危害人类健康的常见病和多发病。全球大约有1.7亿的人口感染丙型肝炎病毒(Hepatitis C Virus，HCV)。我国HCV感染者已达6000万。HCV急性感染后50-80％转变为慢性感染，其中的10-20％发展成肝硬化，并与肝细胞癌的发生密切相关。迄今α-干扰素和病毒唑联合治疗是唯一的治疗方法，但这种治疗方法只对不到50％的患者显效，且具有费用高、易复发和副作用多等缺陷；尤其是我国流行的HCV 1b型，对α干扰素的治疗应答最低，因此，加强对HCV防治的研究至关重要。
HCV为单股正链RNA病毒，属黄病毒科，全长约9600个核苷酸，ORF区编码3010个氨基酸的多聚蛋白前体，经过宿主和病毒本身基因编码的蛋白酶裂解为十个功能性片段，包括4个结构蛋白(核心蛋白C、包膜蛋白E1、E2以及P7)及6个非结构蛋白(NS2、NS3、NS4A、NS4B、NS5A和NS5B)。HCV RNA 5’端没有帽子结构，由5’非编码区(NCR)内的内部核糖体进入位点(IRES)介导启动并控制着整个HCV基因组的翻译过程，而HCV NS3蛋白具有丝氨酸蛋白酶和解旋酶活性，参与病毒蛋白翻译后加工，为病毒复制所必须。因此IRES及HCV NS3／4A丝氨酸蛋白酶是目前抗病毒治疗研究的主要靶位。
多年来针对HCV NS3／4A及HCV IRES抑制剂的研究取得可喜的进展，但由于缺乏理想的小动物模型，这些药物在体内抗HCV的作用效果，以及对其毒性、安全性等难以作出评估，极大地延缓了这些药物进入临床，因而建立适用于HCV IRES和NS3蛋白抑制剂筛选及评价小鼠模型迫在眉睫。
Hepatitis C virus (HCV) represents a major health concern, with the number of HCV-infected individuals estimated to be 170 million worldwide. It becomes persistent among 50 to 80% infected persons, and such patients may develop chronic hepatitis and cirrhosis, some may even progress to the stage of hepatocellular carcinoma.Today's therapies based on IFN-α coupling with ribavirin can induce a sustained virological and biochemical response in less than 50% of treated patients, depending on the virus genotypes, the virus load and the patient's age. So the development of a new way for HCV prevention and treatment is very desirable.
HCV is a blood-borne enveloped RNA-containing member of the Flavivirus family, approximately 9.6 kb in length. It contains a single large open reading frame (ORF). The HCV ORF encodes a polypeptide of about 3010 amino acid residues. This polypeptide has been suggested to be proteolyticaly processed into 10 viral proteins. Among these proteins, C,E1,E2 and P7 were considered to be structure proteins, and NS2,NS3,NS4A,NS4B,NS5A,NS5B non-structure (NS) proteins. The 5' UTR represents the most conserved region of the genome.lt forms the highly structured internal ribosome entry site (IRES),which directs translation in a cap-independent manner. The N terminus of NS3 protein contains a serine protease domain that forms a tight non-covalent complex with the cofactor NS4A. The C-terminal two-thirds of NS3 protein possess helicase and NTPase activity. These vital functions make IRES and NS3 as two potential drug targets in anti-HCV research.
HCV为单股正链RNA病毒，属黄病毒科，全长约9600个核苷酸，ORF区编码3010个氨基酸的多聚蛋白前体，经过宿主和病毒本身基因编码的蛋白酶裂解为十个功能性片段，包括4个结构蛋白(核心蛋白C、包膜蛋白E1、E2以及P7)及6个非结构蛋白(NS2、NS3、NS4A、NS4B、NS5A和NS5B)。HCV RNA 5’端没有帽子结构，由5’非编码区(NCR)内的内部核糖体进入位点(IRES)介导启动并控制着整个HCV基因组的翻译过程，而HCV NS3蛋白具有丝氨酸蛋白酶和解旋酶活性，参与病毒蛋白翻译后加工，为病毒复制所必须。因此IRES及HCV NS3／4A丝氨酸蛋白酶是目前抗病毒治疗研究的主要靶位。
多年来针对HCV NS3／4A及HCV IRES抑制剂的研究取得可喜的进展，但由于缺乏理想的小动物模型，这些药物在体内抗HCV的作用效果，以及对其毒性、安全性等难以作出评估，极大地延缓了这些药物进入临床，因而建立适用于HCV IRES和NS3蛋白抑制剂筛选及评价小鼠模型迫在眉睫。
Hepatitis C virus (HCV) represents a major health concern, with the number of HCV-infected individuals estimated to be 170 million worldwide. It becomes persistent among 50 to 80% infected persons, and such patients may develop chronic hepatitis and cirrhosis, some may even progress to the stage of hepatocellular carcinoma.Today's therapies based on IFN-α coupling with ribavirin can induce a sustained virological and biochemical response in less than 50% of treated patients, depending on the virus genotypes, the virus load and the patient's age. So the development of a new way for HCV prevention and treatment is very desirable.
HCV is a blood-borne enveloped RNA-containing member of the Flavivirus family, approximately 9.6 kb in length. It contains a single large open reading frame (ORF). The HCV ORF encodes a polypeptide of about 3010 amino acid residues. This polypeptide has been suggested to be proteolyticaly processed into 10 viral proteins. Among these proteins, C,E1,E2 and P7 were considered to be structure proteins, and NS2,NS3,NS4A,NS4B,NS5A,NS5B non-structure (NS) proteins. The 5' UTR represents the most conserved region of the genome.lt forms the highly structured internal ribosome entry site (IRES),which directs translation in a cap-independent manner. The N terminus of NS3 protein contains a serine protease domain that forms a tight non-covalent complex with the cofactor NS4A. The C-terminal two-thirds of NS3 protein possess helicase and NTPase activity. These vital functions make IRES and NS3 as two potential drug targets in anti-HCV research.
引文
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