N-（5-取代-1,3,4-噻二唑-2-基）-酰胺类化合物的合成、结构表征及其生物活性研究

为了寻找高效生物活性的化合物,笔者设计合成了一系列同时含有1,3,4-噻二唑、酰胺及芳氧亚甲基等活性基团的化合物.首先以取代羧酸和氨基硫脲为起始原料,三氯氧磷为脱水剂,脱水环化得到2-氨基-5-取代-1,3,4-噻二唑,然后再将所得到的产物与酰氯进行酰胺化反应,合成出12个新的N-(5-取代-1,3,4-噻二唑-2-基)-酰胺类化合物.将所合成的化合物通过元素分析、红外光谱、核磁共振氢谱等测试手段进行分析,确证结构.初步观察了所合成的目标化合物在10 ppm浓度下对小麦胚鞘发芽的影响,测试结果表明,所合成的目标化合物中大部分对小麦芽鞘生长具有抑制作用.     

吡咯[2,3-d]嘧啶核苷单磷酸酯的合成和抗HBV活性

以4-取代-9-(2′-脱氧-2′-β-氟-β-D-呋喃糖基)吡咯[2,3-d]嘧啶类化合物为原料,与三氯氧磷一锅法合成出具有新型结构的吡咯[2,3-d]嘧啶核苷单磷酸酯类化合物,其化学结构经核磁共振磷谱(31P NMR)、核磁共振氢谱(1 H NMR)、高分辨质谱(HRMS)分析确证;对合成的目标化合物进行了初步的体外抗乙肝病毒药理活性实验,结果显示设计合成的7个化合物对乙型肝炎病毒表面抗原(HBsAg)和乙型肝炎e抗原(HBeAg)都有一定的抑制作用,其中化合物3a,3d,3f对HBsAg和HBeAg的抑制率较高,3a的毒性较低.     

芳基磺酰氨基丙基杂氮硅三环化合物的合成及生物活性研究

杂氮硅三环是具有特殊结构和显著生物活性的有机硅化合物．在杂氮硅三环化合物中引入磺酰基,合成了8个芳基磺酰氨基丙基杂氮硅三环化合物,经IR,1HNMR,MS及元素分析确定了它们的结构,初步的生物活性测试表明：所有的目标化合物对金黄色葡萄球菌都有抑制作用,其中活性较好的为侧链胺基氢原子未被完全取代的化合物．     

3-O-甲基-4，6-O-苯亚甲基-N-（2-吡啶基）-α-D-葡萄糖的合成研究

研究了以α-D-葡萄糖为原料，经异丙叉基保护、甲基化、苯亚甲基化等步骤合成了3-O-甲基-4，6-O-苯亚甲基-N-（2-吡啶基）-α-D-葡萄糖，产物经质谱、核磁共振谱及红外光谱对其进行了结构表征分析。N-杂环取代的糖类化合物是一种重要的医药和有机合成中间体，该类化合物将在合成糖基、核苷类似物、杂环化合物、糖多肽等具有潜在药理活性的化合物中具有广阔的应用前景．     

CuI催化的A3-偶联反应合成邻羟基苯基丙炔胺类化合物的研究

丙炔胺类化合物因其具有多个反应位点而在有机合成中具有广泛的应用。本文发展了一种CuI催化的水杨醛、四氢吡咯和末端炔的A³-偶联反应合成邻羟基苯基丙炔胺衍生物的方法。水杨醛和芳基乙炔中无论是吸电子基团还是供电子基团均能较好的适用于该反应，且水杨醛中含有多个取代基时也能得到相应的目标产物，并利用¹H,¹³C NMR和HRMS等手段对目标化合物的结构进行了表征。该合成方法具有操作简单、合成效率高和取代基容忍性好等优点，为邻羟基苯基丙炔胺类化合物的合成提供了一种新的合成思路。     

芳基磺酰氨基丙基杂氮硅三环化合物的合成及生物？…

杂氮硅三环是具有特殊结构和显著生物活性的有机硅化合物，在杂氮硅三环化合物中引入磺酰基，合成了８个芳基磺酰氨基丙基杂氮硅三环化合物，经ＩＲ，＾１ＨＮＭＲ，ＭＳ及元素分析确定了它们的结构，初步的生物活性测试表明：所有的目标化合物对金黄色葡萄球菌都有抑制作用，其中活怀较好的为侧链胺基氨原子未被完全取代的化合物。     

齐多夫定衍生物的合成及抗菌活性研究

二氢叶酸还原酶是细菌存活所必需的酶,且在哺乳动物中的氨基酸序列存在显著差异,DNA结构同源性很小,因此这些酶可以作为抗细菌感染的优秀药物靶标.为了寻找新型抗菌化合物,以齐多夫定为先导物,分别和不同取代基的芳香类的末端炔发生反应,利用药物拼合原理,将活性基团引入到目标产物结构中,得到一系列齐多夫定链接1,2,3-三氮唑的新化合物,结构经~1 H NMR,~(13) C NMR和HRMS确证.对合成化合物进行初步生物活性测试,部分化合物对化脓链球菌显示抑制作用.     

4-氨基-1,2,4-三唑-3-酮席夫碱合成及毒性

以自制的4－氨基－1，2，4－三唑－3－酮为原料，分别与取代的芳香醛反应，采用已有的合成方法设计合成了8个席夫碱类化合物，所有化合物通过^1H NMR,IR，元素分析和熔点测试，确定了化合物的分子结构，初步测定了它们在不同浓度下对金黄色葡萄球菌、大肠杆菌的抑制作用。测试结果表明，部分化合物对金黄色葡萄球菌有较好的抑制作用。     

异脲盐法合成胍及其N-取代嘧啶胺衍生物的研究

采用异脲盐法合成一系列N-取代胍和N-取代-4-(3-吡啶基)嘧啶-2-胺类化合物。以O-甲基异脲硫酸盐为起始原料,与脂肪胺类或芳香胺类化合物反应合成脂肪胍或芳香胍类化合物(1),再以3-乙酰吡啶和N,N-二甲基甲酰胺二甲基缩醛(DMF-DMA)为原料反应合成3-(二甲基氨基)-1-(3-吡啶基)-2-丙烯-1-酮(2),将化合物(1)和(2)在碱性条件下反应,最终得到目标产物N-取代-4-(3-吡啶基)嘧啶-2-胺(3)。合成出了6个N-取代胍及其相应的衍生物N-取代-4-(3-吡啶基)嘧啶-2-胺类化合物,并用1H NMR,IR和元素分析表征了目标产物的结构。该方法路线简捷环保、原料廉价易得、反应条件较温和。     

含氰基功能基的半乳糖苷衍生物的合成研究

利用哺乳动物肝细胞的特异性受体可使半乳糖基化后的药物定向地转运至肝细胞，从而获得肝脏摄取率高的药物。而合成此类药物的关键在于合成含功能基的化学配体。以易得的乙二醇为起始原料，经加成取代以及缩合三步反应，成功地得到含氰基功能基的半乳糖苷衍生物(1)，其结构经^1HNMR、IR、MS确证，实验结果表明此方法是合成目标化合物的较好方法。     

PMNBP缩氨基酸席夫碱配合物的合成、表征和晶体结构

合成1-苯基-3-甲基-4-对硝基苯甲酰基-5-吡唑啉酮（PMNBP）缩苯丙氨酸甲酯席夫碱及其Ni（Ⅱ）、Co（Ⅱ）金属配合物,并对合成的配合物进行红外光谱的表征.利用X射线单晶衍射法测定配合物的晶体结构,结果表明二者均为六配位同构配合物.     

An NS3 protease inhibitor with antiviral effects in humans infected with hepatitis C virus

Hepatitis C virus (HCV) infection is a serious cause of chronic liver disease worldwide with more than 170 million infected individuals at risk of developing significant morbidity and mortality. Current interferon-based therapies are suboptimal especially in patients infected with HCV genotype 1, and they are poorly tolerated, highlighting the unmet medical need for new therapeutics. The HCV-encoded NS3 protease is essential for viral replication and has long been considered an attractive target for therapeutic intervention in HCV-infected patients. Here we identify a class of specific and potent NS3 protease inhibitors and report the evaluation of BILN 2061, a small molecule inhibitor biologically available through oral ingestion and the first of its class in human trials. Administration of BILN 2061 to patients infected with HCV genotype 1 for 2 days resulted in an impressive reduction of HCV RNA plasma levels, and established proof-of-concept in humans for an HCV NS3 protease inhibitor. Our results further illustrate the potential of the viral-enzyme-targeted drug discovery approach for the development of new HCV therapeutics.     

A genetically humanized mouse model for hepatitis C virus infection

Hepatitis C virus (HCV) remains a major medical problem. Antiviral treatment is only partially effective and a vaccine does not exist. Development of more effective therapies has been hampered by the lack of a suitable small animal model. Although xenotransplantation of immunodeficient mice with human hepatocytes has shown promise, these models are subject to important challenges. Building on the previous observation that CD81 and occludin comprise the minimal human factors required to render mouse cells permissive to HCV entry in vitro, we attempted murine humanization via a genetic approach. Here we show that expression of two human genes is sufficient to allow HCV infection of fully immunocompetent inbred mice. We establish a precedent for applying mouse genetics to dissect viral entry and validate the role of scavenger receptor type B class I for HCV uptake. We demonstrate that HCV can be blocked by passive immunization, as well as showing that a recombinant vaccinia virus vector induces humoral immunity and confers partial protection against heterologous challenge. This system recapitulates a portion of the HCV life cycle in an immunocompetent rodent for the first time, opening opportunities for studying viral pathogenesis and immunity and comprising an effective platform for testing HCV entry inhibitors in vivo.     

Claudin-1 is a hepatitis C virus co-receptor required for a late step in entry

Hepatitis C virus (HCV) is a leading cause of cirrhosis and liver cancer worldwide. A better understanding of the viral life cycle, including the mechanisms of entry into host cells, is needed to identify novel therapeutic targets. Although HCV entry requires the CD81 co-receptor, and other host molecules have been implicated, at least one factor critical to this process remains unknown (reviewed in refs 1-3). Using an iterative expression cloning approach we identified claudin-1 (CLDN1), a tight junction component that is highly expressed in the liver, as essential for HCV entry. CLDN1 is required for HCV infection of human hepatoma cell lines and is the first factor to confer susceptibility to HCV when ectopically expressed in non-hepatic cells. Discrete residues within the first extracellular loop (EL1) of CLDN1, but not protein interaction motifs in intracellular domains, are critical for HCV entry. Moreover, antibodies directed against an epitope inserted in the CLDN1 EL1 block HCV infection. The kinetics of this inhibition indicate that CLDN1 acts late in the entry process, after virus binding and interaction with the HCV co-receptor CD81. With CLDN1 we have identified a novel key factor for HCV entry and a new target for antiviral drug development.     

基于微小发光真菌的化妆品化学毒性测试方法

为有效对化妆品化学毒性进行可靠性测试,研究了一种基于微小磁微粒化学发光真菌的化妆品化学毒性准确测试方法。化妆品隐藏毒性严重危害人类健康,在人群中的易感率大;所以对化妆品毒性的准确测试至关重要。传统的化妆品毒性检测方法采用基于毒性反应机理的化学测试方法。通过化妆品毒性中特有的毒性物质与相关的化学物质发生反应,生成一定的反应产物或者降解产物来检测化妆品毒性是否存在;此方法在以往的检测中达到了一定的效果。但是随着毒性变种的逐步扩散,化妆品毒性多样化逐渐形成,传统的测试方法不能对毒性变种扩散有效检测。提出一种基于磁微粒化学发光法的微小发光真菌化妆品毒性可靠测试方法。通过磁微粒发光原理来检测化妆品毒性的存在,通过化学实验,用磁微粒化学反应形成稳定的检测物,形成稳定的化妆品毒性检测效果和结论。测试方法简单易行、可靠性高,所以具有很好的实践应用价值。     

HCV多表位抗原基因的表达及其在小鼠中诱导的抗体应答研究

丙型肝炎病毒(HCV)是引起病毒性非甲非乙肝炎的主要病原．全世界约有1．7亿HCV的感染者,我国估计占4000万以上．发展有效的丙型肝炎疫苗刻不容缓．构建了3段串联重复的丙型肝炎病毒多表位抗原基因3如,将其克隆到His融合表达载体pET-28b( )上后在大肠杆菌BL21(DE3)中得到了高效表达．3PCX经Ni^2 -NTA—agarose柱纯化后免疫BALB／c小鼠,诱发了高水平的抗体免疫应答．     

Evasion of intracellular host defence by hepatitis C virus

Viral infection of mammalian cells rapidly triggers intracellular signalling events leading to interferon alpha/beta production and a cellular antiviral state. This 'host response' is our first line of immune defence against infection as it imposes several barriers to viral replication and spread. Hepatitis C virus (HCV) evades the host response through a complex combination of processes that include signalling interference, effector modulation and continual viral genetic variation. These evasion strategies support persistent infection and the spread of HCV. Defining the molecular mechanisms by which HCV regulates the host response is of crucial importance and may reveal targets for novel therapeutic strategies.     

SDS在HCV基因工程抗原包被中的应用研究

在间接ELISA法检测HCV抗体过程中,通过使用含有十二烷基硫酸钠（SDS）的包被液对HCV基因工程抗原固相化,发现使用SDS提高了HCV间接ELISA法诊断试剂检测的阳性标本和阴性标本的偏比,改善了反应模式,为改善HCV-ELISA法诊断试剂的质量提供了有效途径.     

Modelling how ribavirin improves interferon response rates in hepatitis C virus infection

Nearly 200 million individuals worldwide are currently infected with hepatitis C virus (HCV). Combination therapy with pegylated interferon and ribavirin, the latest treatment for HCV infection, elicits long-term responses in only about 50% of patients treated. No effective alternative treatments exist for non-responders. Consequently, significant efforts are continuing to maximize response to combination therapy. However, rational therapy optimization is precluded by the poor understanding of the mechanism(s) of ribavirin action against HCV. Ribavirin alone induces either a transient early decline or no decrease in HCV viral load, but in combination with interferon it significantly improves long-term response rates. Here we present a model of HCV dynamics in which, on the basis of growing evidence, we assume that ribavirin decreases HCV infectivity in an infected individual in a dose-dependent manner. The model quantitatively predicts long-term response rates to interferon monotherapy and combination therapy, fits observed patterns of HCV RNA decline in patients undergoing therapy, reconciles conflicting observations of the influence of ribavirin on HCV RNA decline, provides key insights into the mechanism of ribavirin action against HCV, and establishes a framework for rational therapy optimization.     

Specific immune responses induced by a multi-epitope antigen of hepatitis C virus in mice and rabbits

Five highly conserved and immunogenic epitopes of hepatitis C virus (HCV) have been chosen to form a multi-epitope antigen gene and fused with β-galactosidase gene to express a hybrid GZ-PCX antigen, which could be specifically recognized by human HCV sera. High level of anti-GZ-PCX IgG has been induced when mice or rabbits were immunized with GZ-PCX antigen emulsificated with complete Freund’s adjuvant or mixed with killed attenuatedSalmonella typhimurium SL3261. The specific anti-GZ-PCX IgG reached a high titer of 10^-6, which remained for several months. Specific cytotoxic T lymphocyte (CTL) effects, delayed type hypersensitivity reaction (DTH) and proliferation of peripheral lymphocytes have been induced by GZ-PCX antigen or synthetic peptides. High level of anti-GZ-PCX slgG has been detected in mice’s intestinal washing fluids, which indicates that the antigen induced mucosal immunity as well as systematic immunity. The studies show that the HCV multi-epitope antigen induces high level of specific immune responses without obvious toxicity, which might be able to provide protectivity to any HCV genotypes and isolates.