Global stability and periodic solution of the viral dynamics

It is well known that the mathematical models provide very important information for the research of human immunodeficiency virus-type 1 and hepatitis C virus (HCV). However, the infection rate of almost all mathematical models is linear. The linearity shows the simple interaction between the T cells and the viral particles. In this paper, we consider the classical mathematical model with saturation response of the infection rate. By stability analysis we obtain sufficient conditions on the parameters for the global stability of the infected steady state and the infection-free steady state. We also obtain the conditions for the existence of an orbitally asymptotically stable periodic solution. Numerical simulations are presented to illustrate the results.     

Syntheses of nucleosides with 2′-spirolactam and 2′-spiropyrrolidine moieties as potential inhibitors of hepatitis C virus NS5B polymerase

To discover novel nucleosides as potential antiviral agents, 2′-spirolactam and 2′-spiropyrrolidine-containing nucleoside analogs were envisioned. Efficient synthetic routes were developed with an epoxide opening as the key step to establish the quaternary center at the 2′ position, leading to the design and synthesis of uridine analogs 8 and 21, prodrugs 13–16, and cytidine analog 11.     

Syntheses of 4′-spirocyclic phosphono-nucleosides as potential inhibitors of hepatitis C virus NS5B polymerase

To discover novel nucleosides as potential antiviral agents, 4′-spirocyclic phosphono-nucleosides were designed to mimic the monophosphate of R-1479, a known nucleoside inhibitor of HCV NS5B. Bypassing the first kinase step to nucleoside monophosphate is viewed as advantageous since this phosphorylation is often observed as the rate-limiting transformation to the active NTP for many nucleosides. Efficient synthetic routes were developed with a triphenylphosphine–iodine cyclization reaction as the key step to form the tetrahydrofuran 4′-spirocycle. The desired 4′-spirocyclic phosphono-cytidine analogs 12a, 12b, and 16 were prepared in 11 steps.     

Inhibitor Development against p7 Channel in Hepatitis C Virus

Hepatitis C Virus (HCV) is the key cause of chronic and severe liver diseases. The recent direct-acting antiviral agents have shown the clinical success on HCV-related diseases, but the rapid HCV mutations of the virus highlight the sustaining necessity to develop new drugs. p7, the viroporin protein from HCV, has been sought after as a potential anti-HCV drug target. Several classes of compounds, such as amantadine and rimantadine have been testified for p7 inhibition. However, the efficacies of these compounds are not high. Here, we screened some novel p7 inhibitors with amantadine scaffold for the inhibitor development. The dissociation constant (Kd) of 42 ARD-series compounds were determined by nuclear magnetic resonance (NMR) titrations. The efficacies of the two best inhibitors, ARD87 and ARD112, were further confirmed using viral production assay. The binding mode analysis and binding stability for the strongest inhibitor were deciphered by molecular dynamics (MD) simulation. These ARD-series compounds together with 49 previously published compounds were further analyzed by molecular docking. Key pharmacophores were identified among the structure-similar compounds. Our studies suggest that different functional groups are highly correlated with the efficacy for inhibiting p7 of HCV, in which hydrophobic interactions are the dominant forces for the inhibition potency. Our findings provide guiding principles for designing higher affinity inhibitors of p7 as potential anti-HCV drug candidates.     

Chromatographic analysis of ledipasvir and sofosbuvir: New treatment for chronic hepatitis C infection with application to human plasma

Sofosbuvir (SOF) and ledipasvir (LED) are recently approved and coformulated as directly acting antiviral agents used for treatment of hepatitis C virus (HCV). A reversed phase high performance liquid chromatography - diode array detector (RP-HPLC/DAD) method was developed and validated for the first time for the analysis of newly formulated anti-HCV combination, in pure form, pharmaceutical formulation and in human plasma. In the developed method, separation was performed on Zorbax® Eclipse C18 column using a gradient mixture of acetonitrile–water as a mobile phase and scanning was performed at 260?nm (for SOF) and 330?nm (for LED). The two drugs were completely separated from each other and from plasma, where plasma peak appeared at 2.76?±?0.05?min, SOF at 4.25?±?0.05, and LED at 7.35?±?0.05. The developed method showed high sensitivity, the drugs showed linearity in the range of 1–45?µg/mL for both pure form and spiked human plasma. Three freeze–thaw cycles were performed separately at two different temperatures, ?8 and ?20°C. No significant loss of the studied drugs were observed during repeated thawing and freezing. Validation parameters such as accuracy, precision, robustness, and ruggedness were tested in compliance with USP recommendations, where acceptable results were obtained. Applying to pharmaceutical formulation showed no interference from tablet excipients.     

Synthetic approaches to imino sugar (2R,3R,4S)-2-(hydroxymethyl)-4-methylpyrrolidine-3,4-diol from naturally occurring sugar and amino acid

Imino sugar compound 3 was prepared by two alternative routes starting from ribose and d-serine. From d-serine (3R,4R,5R)-3,4-dihydroxy-5-(hydroxymethyl)-3-methylpyrrolidin-2-one was prepared in five steps in 50% overall yield, which was further converted into (2R,3R,4S)-2-(hydroxymethyl)-4-methylpyrrolidine-3,4-diol in three steps in 23% overall yield.     

离心力和滑翔距离对于高超声速巡航飞行性能的影响

高超声速巡航飞行器(HCV)是21世纪国际空天技术竞争的焦点, 它的飞行性能可以用初始发射质量与广义有效载荷之比衡量. 文献[1]建立了该性能的耦合分析框架. 在这个框架下进一步考虑了离心力和滑翔距离两个重要因素的影响. 定量分析表明: 给定飞行距离, 对HCV飞行性能影响最大的是巡航速度, 其次是升阻比, 再次是巡航发动机比冲. 随着飞行距离的增加, 由于离心力和滑翔距离的计入, 火箭动力HCV的飞行性能大幅提高, 从近中程到远程均显著优于经典的最小能量弹道. 吸气式冲压发动机尽管比冲大幅优于火箭发动机, 但在巡航速度适用范围和飞行器升阻比方面, 均不及后者. 综合而言, 火箭动力HCV是现阶段非常有竞争力的选择.      

非核苷类HCV NS5B聚合酶抑制剂

全球约有1.7亿人感染丙型肝炎病毒(HCV),至今尚无有效的疫苗和治疗药物。目前干扰素-α和利巴韦林联用抗HCV 的标准治疗方案存在严重不良反应。近年来,新型抗HCV的研究主要集中于针对病毒感染的特异性靶点寻找新的抑制剂。NS5B聚合酶因其在HCV RNA基因组复制中的关键性作用,是目前抗HCV药物设计的重要靶点之一。NS5B聚合酶抑制剂可分为核苷类抑制剂(NIs)和非核苷类抑制剂(NNIs)。自从第一个NS5B NNIs苯并咪唑类化合物被报道以来,大量不同结合位点NS5B NNIs相继被报道,一些化合物在临床研究中表现出了可喜的疗效。本文综述了苯并噻二嗪类、苯并咪唑类、吲哚类、噻吩羧酸类、喹啉酮类、二氢吡喃类和二酮酸类等 NS5B NNIs的研究进展,旨在为该类抑制剂的开发研究提供参考。     

HCVNS2基因表达载体的构建及在E.coli中表达

PCR人含有丙肝病毒全长非结构蛋白的载体pBlueBac25中扩增出全长的NS2基因DNA片段,分别克隆到表达载体pQE30和转座载体pFasBacHTb的多隆位点（MCS）,PFastNS2通过转座插入穿梭载体Bacmid的表达盒;pQENS2转化JM109菌株,诱导表达出N端含有6个His的全长His的全长NS2蛋白,用Ni-NTA-agarose柱层纯化,获得提纯的全长NS2蛋白。