Azetidines and spiro azetidines as novel P2 units in hepatitis C virus NS3 protease inhibitors

Herein, we report the synthesis and structure–activity relationship studies of new analogs of boceprevir 1 and telaprevir 2. Introduction of azetidine and spiroazetidines as a P2 substituent that replaced the pyrrolidine moiety of 1 and 2 led to the discovery of a potent hepatitis C protease inhibitor 37c (EC₅₀ = 0.8 μM).     

Structure–activity studies of (−)-epigallocatechin gallate derivatives as HCV entry inhibitors

Preventing viral entry into cells is a recognized approach for HIV therapy and has attracted attention for use against the hepatitis C virus (HCV). Recent reports described the activity of (−)-epigallocatechin gallate (EGCG) as an inhibitor of HCV entry with modest potency. EGCG is a polyphenolic natural product with a wide range of biological activity and unfavorable pharmaceutical properties. In an attempt to identify more drug-like EGCG derivatives with improved efficacy as HCV entry inhibitors, we initiated structure–activity investigations using semi-synthetic and synthetic EGCG analogs. The data show that there are multiple regions in the EGCG structure that contribute to activity. The gallate ester portion of the molecule appears to be of particular importance as a 3,4-difluoro analog of EGCG enhanced potency. This derivative and other active compounds were shown not to be cytotoxic in Huh-7 cell culture. These data suggest that more potent, non-cytotoxic EGCG analogs can be prepared in an attempt to identify more drug-like candidates to treat HCV infection by this mechanism.     

Synthesis and evaluation of Janus type nucleosides as potential HCV NS5B polymerase inhibitors

The synthesis of new ribo and 2′-β-C-methyl ribo Janus type nucleosides J-AA, J-AG and J-AU is reported along with their ability to block HCV and HIV replication. Their toxicity was also assessed in Huh7, human lymphocytes, CEM and Vero cells.     

Hepatitis C NS5B polymerase inhibitors: 4,4-Dialkyl-1-hydroxy-3-oxo-3,4-dihydronaphthalene-3-yl benzothiadiazine derivatives

4,4-Dialkyl-1-hydroxy-3-oxo-3.4-dihydronaphthalene-3-yl benzothiadiazine derivatives were synthesized and evaluated as inhibitors of genotypes 1a and 1b HCV NS5B polymerase. A number of these compounds exhibited potent activity against genotypes 1a and 1b HCV polymerase in both enzymatic and cell culture activities. A representative compound also showed favorable pharmacokinetics in the rat.     

Could 5′-N and S ProTide analogues work as prodrugs of antiviral agents?

The nucleoside/nucleotide derived antiviral agents have been the most important components of antiviral therapy used in clinics. Recently, the focus of the medicinal chemists within this exciting research field has been affected mainly by the lack of effective therapies for the Hepatitis C virus (HCV) infection and several other “neglected” diseases caused by viruses such as Zika or Dengue. 2′-Methyl modified nucleosides and their monophosphate prodrugs (ProTides) have revolutionized the therapies for HCV in the last few years and, according to the latest research efforts, have also brought a promise for treatment of diseases caused by other members of Flaviviridae family. Here, we report on the design and synthesis of 5’-N and S modified ProTides derived from 2′-methyladenosine. We studied potential applicability of these derivatives as prodrugs of this archetypal antiviral compound.     

Hemin potentiates the anti-hepatitis C virus activity of the antimalarial drug artemisinin

We report that the antimalarial drug artemisinin inhibits hepatitis C virus (HCV) replicon replication in a dose-dependent manner in two replicon constructs at concentrations that have no effect on the proliferation of the exponentially growing host cells. The 50% effective concentration (EC(50)) for inhibition of HCV subgenomic replicon replication in Huh 5-2 cells (luciferase assay) by artemisinin was 78+/-21 microM. Hemin, an iron donor, was recently reported to inhibit HCV replicon replication [mediated by inhibition of the viral polymerase (C. Fillebeen, A.M. Rivas-Estilla, M. Bisaillon, P. Ponka, M. Muckenthaler, M.W. Hentze, A.E. Koromilas, K. Pantopoulos, Iron inactivates the RNA polymerase NS5B and suppresses subgenomic replication of hepatitis C virus, J. Biol. Chem. 280 (2005) 9049-9057.)] at a concentration that had no adverse effect on the host cells. When combined, artemisinin and hemin resulted, over a broad concentration range, in a pronounced synergistic antiviral activity. Also at a concentration (2 microM) that alone had no effect on HCV replication, hemin still potentiated the anti-HCV activity of artemisinin.     

Synthesis and Antiviral Evaluation of Ribavirin Congeners Containing a Hexitol Moiety

Abstract

Several ribavirin congeners containing a hexitol moiety were prepared via ring opening of two different epoxides with the methylcarboxylate ester of triazole and further elaboration. Unfortunately, none of the newly synthesized compounds displayed appreciable antiviral activity.     

Discovery of pyrido[2,3-d]pyrimidine-based inhibitors of HCV NS5A

Efforts to improve the genotype 1a potency and pharmacokinetics of earlier naphthyridine-based HCV NS5A inhibitors resulted in the discovery of a novel series of pyrido[2,3-d]pyrimidine compounds, which displayed potent inhibition of HCV genotypes 1a and 1b in the replicon assay. SAR in this system revealed that the introduction of amides bearing an additional ‘E’ ring provided compounds with improved potency and pharmacokinetics. Introduction of a chiral center on the amide portion resulted in the observation of a stereochemical dependence for replicon potency and provided a site for the attachment of functional groups useful for improving the solubility of the series. Compound 21 was selected for administration in an HCV-infected chimpanzee. Observation of a robust viral load decline provided positive proof of concept for inhibition of HCV replication in vivo for the compound series.     

MK-8325: A silyl proline-containing NS5A inhibitor with pan-genotype activity for treatment of HCV

HCV NS5A inhibitors have shown impressive in vitro potency profiles in HCV replicon assays thus making them attractive components for inclusion in an all oral fixed dose combination regimen. Herein, we describe the discovery and characterization of silyl proline-containing HCV NS5A inhibitor MK-8325 with good pan-genotype activity and acceptable pharmacokinetic properties.     

Synthesis and evaluation of 2′-dihalo ribonucleotide prodrugs with activity against hepatitis C virus

1. Download : Download high-res image (149KB)
2. Download : Download full-size image

Hepatitis C virus (HCV) nucleoside inhibitors have been a key focus of nearly 2 decades of HCV drug research due to a high barrier to drug resistance and pan-genotypic activity profile provided by molecules in this drug class. Our investigations focused on several potent 2′-halogenated uridine-based HCV polymerase inhibitors, resulting in the discovery of novel 2′-deoxy-2′-dihalo-uridine analogs that are potent inhibitors in replicon assays for all genotypes. Further studies to improve in vivo performance of these nucleoside inhibitors identified aminoisobutyric acid ethyl ester (AIBEE) phosphoramidate prodrugs 18a and 18c, which provide high levels of the active triphosphate in dog liver. AIBEE prodrug 18c was compared with sofosbuvir (1) by co-dosing both compounds by oral administration in dog (5 mg/kg each) and measuring liver concentrations of the active triphosphate metabolite at both 4 and 24 h post dosing. In this study, 18c provided liver triphosphate concentrations that were 6-fold higher than sofosbuvir (1) at both biopsy time points, suggesting that 18c could be a highly effective agent for treating HCV infected patients in the clinic.     

A new natural α-helical peptide from the venom of the scorpion Heterometrus petersii kills HCV

Hepatitis C virus (HCV) is a major cause of chronic liver disease, cirrhosis, and hepatocellular carcinoma. There is no vaccine available for HCV, and almost half of patients cannot be cured using standard combination therapy. Thus, new anti-HCV strategies and drugs are urgently needed. Here, the gene encoding a new α-helical peptide, Hp1090, was screened from the venomous gland cDNA library of the scorpion Heterometrus petersii. Structural analysis showed that Hp1090 is an amphipathic α-helical peptide. In vitro HCV RNA inhibitory assays indicated that Hp1090 peptide inhibited HCV infection with an IC₅₀ of 7.62 μg/ml (5.0 μM), whereas Hp1035 peptide, showing high homology to Hp1090, exhibited no anti-HCV activity. Hp1090 acted as a viricide against HCV particles in vitro and prevented the initiation of HCV infection. Furthermore, this peptide interacted with HCV particles directly and rapidly permeabilized phospholipid membranes. Collectively, it seems that Hp1090 is virocidal for HCV in vitro, directly interacting with the viral membrane and decreasing the virus infectivity. These results suggest that Hp1090 could be considered an anti-HCV lead compound with virocidal mechanism that offers a potential therapeutic approach to HCV infection. Our work opens a new avenue for antiviral drug discovery in natural scorpion venom.     

Mathematical modelling of hepatitis C treatment for injecting drug users

Hepatitis C virus (HCV) is a blood-borne infection that can lead to progressive liver failure, cirrhosis, hepatocellular carcinoma and death. In developed countries, the majority of HCV infections are transmitted via injecting drug users (IDUs). Despite effective antiviral treatment for HCV, very few active IDUs are treated. Reluctance to treat is partially due to the risk of reinfection. We develop a mathematical model of HCV transmission amongst active IDUs, and examine the potential effect of antiviral treatment. As most mathematical models of interventions utilise a treatment function proportional to the infected population, but many policy implementations set fixed yearly targets for specific numbers treated, we study the effects of using two different treatment terms: annually treating a proportion of infecteds or a fixed number of infecteds. We examine the behaviour of the two treatment models and find different bifurcation behaviours in each case. We calculate analytical solutions for the treatment level needed for disease clearance or control, and observe that achievable levels of treatment can result in control or eradication across a wide range of prevalence levels. Finally, we calculate the sensitivity of the critical treatment threshold to the model parameters, and find that for a given observed prevalence, the injecting duration and infection risk play the most important role in determining the treatment level needed. By contrast, the sensitivity analysis indicates the presence (or absence) of immunity does not alter the treatment threshold. We conclude by discussing the public health implications of this work, and comment on the importance and feasibility of utilising treatment as prevention for HCV spread amongst IDUs.     

2012‒2014年杭州市HBsAg、HCVAb、HIV及TP感染情况分析

目的 统计分析2012年～2014年浙江大学医学院附属第一医院就诊患者中HBsAg、HCV、HIV及TP感染情况,了解期间来院患者四种病原体的感染情况,为临床安全提供参考依据。方法 回顾性分析2012年～2014年我院住院患者血清中HBV、HCV、HIV和TP感染状况。结果 2012年至2014年共统计214 935人次,HBsAg阳性率为13.08%,HCV阳性率为0.51%,HIV阳性率为0.37%,TP阳性率为2.44%。各年份间HBsAg差异有统计学意义（HbsAg：χ2=55.74,P<0.0001;HCV：χ2=2.24,P=0.327;HIV：χ2=2.67,P=0.26;TP：χ2=2.65,P=0.27）。不同性别之间,HBsAg、HIV或TP阳性率差异有统计学意义（HBsAg：χ2=2680.6,P<0.0001;HCV：χ2=2.69,P=0.101;HIV：χ2=344.3,P<0.0001;TP：χ2=192.4,P<0.0001）;不同年龄段之间,HBsAg、HCV、HIV及TP阳性率差异有统计学意义（HbsAg：χ2=3036.1,P<0.0001;HCV：χ2=59.45,P<0.0001;HIV：χ2=246.06,P<0.0001;TP：χ2=636.80,P<0.0001）。结论 开展术前四项检测,有助于及时了解患者情况,明确人群感染情况,对制定感染的预防及治疗措施,避免院内感染的发生具有十分重要的意义。     

骨科假体表面涂层技术现状

由植入物界面处的相互作用可能引起无菌性松动和假体周围感染。而无菌性松动和假体周围感染仍然是一个难以治疗的问题,并且最终可能导致假体植入失败,引起严重后果。理想的植入物应能促进骨整合,防止细菌粘附,减少细菌感染。骨科植入技术主要基于生物材料的开发和使用,随着材料科学和细胞生物学的发展,已可以用新的植入物表面涂层的进展来解决这些问题。本文回顾总结了时下骨科常见的假体涂层设计和相关问题,以期为进一步研究提供借鉴。     

江苏省300例艾滋病人群联合检测 HCV、TP、HIV的结果分析

摘要 目的：探究江苏省2017年至2019年度艾滋病哨点监测人群丙型肝炎病毒（Hepatitis C virus,HCV）、梅毒螺旋体（Treponema Pallidum,TP）以及人类免疫缺陷病毒（Treponema Pallidum,HIV）感染状况,以期为制定对应的疾患干预手段提供临床数据支撑。方法：严格按照《全国艾滋病哨点监测实施方案操作手册（2012版）》对入组的900例个体开展相关问卷调查,并对入组对象进行HCV、TP以及HIV感染血清学检测,分别统计3年总体感染情况、年度感染情况、不同年龄段人群感染情况以及合并感染情况。结果：（1）性工作者HCV、TP以及HIV感染率分别为3.00 %、4.00 %和9.00 %,吸毒人群感染率分别为13.00 %、73.00 %和13.00 %,孕产妇感染率分别为1.00 %、0.67 %和1.33 %,比较显示,吸毒人群HCV、TP以及HIV感染率明显高于其余两类人群（P<0.05）;（2）就各年度不同人群HCV、TP以及HIV感染率开展年度横向比较显示,不同年度不同人群的HCV、TP以及HIV感染率差异无统计学意义（P>0.05）;（3）针对不同年龄段不同人群的HCV、TP以及HIV感染率开展比较显示,吸毒人群中≥50岁HCV和HIV感染率明显高于其他年龄段（P<0.05）,性工作者中18~20岁HIV感染率明显高于其他年龄段（P<0.05）,其余差异无统计学意义（P>0.05）;（4）合并感染情况分析显示合并TP以及HIV感染率要高于其他合并感染。结论：吸毒人群是江苏省HCV、TP以及HIV感染高发群体,以≥50岁年龄段感染率最高,总体来看HCV、TP以及HIV感染率呈现逐年降低趋势,但仍应该加强对上述疾患的防控工作。