Discovering novel anti-HCV compounds with inhibitory activities toward HCV NS3/4A protease

Yu, Yongsheng; Jing, Jingfeng; Tong, Xiaowei; He, Pei‐Lan; Li, Yuanchao; Hu, Youhong; Tang, Wei; Zuo, Jian‐Ping

doi:10.1038/aps.2014.55

Cited by 13 publications

(8 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Natural extracts from plants used in Chinese folk medicine have various bioactivities and immunomodulatory effects and provide a promising path for autoimmune hepatitis treatment. Previous studies have identified a large number of substances derived from those plants; these substances have been shown to possess therapeutic potential and low toxicity for treatment of mul- tiple diseases [29,30] . For example, the species Hedyotis, which is used in folk medicine, provides an abundant source of natural products that are beneficial to hepatitis [4] .…”

Section: Discussionmentioning

confidence: 99%

Betulin from Hedyotis hedyotidea ameliorates concanavalin A-induced and T cell-mediated autoimmune hepatitis in mice

et al. 2016

View full text Add to dashboard Cite

Hedyotis hedyotidea has been used in traditional Chinese medicine for the treatment of autoimmune diseases. However, the mechanisms underlying for the effect remain unknown. We previously showed that, among 11 compounds extracted from H hedyotidea, betulin produced the strongest suppressive effect on T cell activation. Here, we examined the hepatoprotective effects of betulin against acute autoimmune hepatitis in mice and the mechanisms underlying the effects. Freshly isolated mouse splenocytes were stimulated with concanavalin A (Con A, 5 μg/mL) in the presence of betulin, the cell proliferation was assessed with CSFE-dilution assay. Mice were injected with betulin (10, 20 mg· kg -1 ·d -1 , ip) for 3 d. One hour after the last injection, the mice were injected with Con A (15 mg/kg, iv) to induce acute hepatitis. Blood samples and liver tissues were harvested at 10 h after Con A injection, and serum transaminase levels and liver histopathology were detected; serum levels of proinflammatory cytokines, hepatic T lymphocyte ratios, and functional statuses of conventional T and NKT cells were also analyzed. Betulin (16 and 32 μmol/L) dose-dependently suppressed the proliferation of Con A-stimulated mouse splenocytes in vitro. In Con A-challenged mice, preinjection with betulin (20 mg· kg -1 ·d -1 ) significantly decreased the levels of proinflammatory cytokines IFN-γ, TNF-α and IL-6, and ameliorated liver injury. Furthermore, pretreatment with betulin (20 mg· kg -1 ·d -1 ) significantly inhibited the Con A-induced activation of NKT and conventional T cells, and decreased production of proinflammatory cytokines IFN-γ, TNF-α and IL-6 in these two cell populations. Betulin has immunomodulatory effect on overly activated conventional T and NKT cells and exerts hepatoprotective action in mouse autoimmune hepatitis. The findings provide evidence for the use of H hedyotidea and its constituent betulin in the treatment of autoimmune diseases.

show abstract

Section: Discussionmentioning

confidence: 99%

Betulin from Hedyotis hedyotidea ameliorates concanavalin A-induced and T cell-mediated autoimmune hepatitis in mice

et al. 2016

View full text Add to dashboard Cite

show abstract

“…The biological assays were performed following a previously reported procedure [ 23 ]. The Huh7.5.1 cell line and the HCV infectious virus J399EM were kindly provided by Xin-Wen Chen, Wuhan Institute of Virology, Chinese Academy of Sciences.…”

Section: Methodsmentioning

confidence: 99%

Discovery of Metal Ions Chelator Quercetin Derivatives with Potent Anti-HCV Activities

et al. 2015

View full text Add to dashboard Cite

Analogues or isosteres of α,γ-diketoacid (DKA) 1a show potent inhibition of hepatitis C virus (HCV) NS5B polymerase through chelation of the two magnesium ions at the active site. The anti-HCV activity of the flavonoid quercetin (2) could partly be attributed to it being a structural mimic of DKAs. In order to delineate the structural features required for the inhibitory effect and improve the anti-HCV potency, two novel types of quercetin analogues, 7-O-arylmethylquercetins and quercetin-3-O-benzoic acid esters, were designed, synthesized and evaluated for their anti-HCV properties in cell-based assays. Among the 38 newly synthesized compounds, 7-O-substituted derivative 3i and 3-O-substituted derivative 4f were found to be the most active in the corresponding series (EC50 = 3.8 μM and 9.0 μΜ, respectively). Docking studies suggested that the quercetin analogues are capable of establishing key coordination with the two magnesium ions as well as interactions with residues at the active site of HCV NS5B.

show abstract

“…To date, quinazoline moiety has been found in several approved drugs such as Gefitinib, Erlotinib and Prazosin which the first and second both served as an anticancer agent while the latter is used to treat anxiety and hypertension [12]. In addition to the aforementioned pharmaceutical values, quinazoline derivatives also being widely studied for other biological significances, such as anti-inflammatory, anti-tubercular anti-HIV, and anti-serine protease activities [11, 13 - 16]. In this study, we have demonstrated the inhibitory potential of five unique quinazoline derivatives towards HCV NS3-4A serine protease and their ability to inhibit HCV replication in a cell-based HCV replicon system.…”

Section: Introductionmentioning

confidence: 99%

Novel Quinazoline derivatives inhibited HCV Serine protease and viral replication in Huh-7 cells

et al. 2019

Preprint

View full text Add to dashboard Cite

1 Drugs against HCV infection are facing several drawbacks such as undesirable side effects, 2 emerging of HCV resistant strains, and high cost of the entire course of treatment. Thus, 3 new active and cost-effective compounds are required to develop effective drugs to combat 4 HCV infection. This study was designed to test the antiviral activity of quinazoline derivatives 5 against HCV infection using HCV protease assay (HCV NS3-4Apro) and cell-based HCV 6 replicon assay. The results showed that some of quinazoline derivatives inhibited HCV NS3-7 4Apro with IC 50 ranged from 42 µM of compound 4 to 150 µM of compound 2. In this study, 8 compound 4 was considered as the best compound lead to developing potent anti-HCV 9 NS3-4Apro inhibitors. The toxic dose of compound 4 was more than 80 µM for 24, 48, and 10 72 h. Compound 4 showed dose-dependent inhibition against HCV replication with a 11 considerable reduction in Rluc activity at 40 µM. This finding was further investigated by 12 immunostaining that showed the inhibitory effect of compound 4 against HCV NS3-4Apro 13 was dose-dependent. This study identified a unique small molecule compound that could 14 lead to the development of HCV NS3-A4pro inhibitors and would be useful to develop highly 15 effective drugs for HCV infection based on HCV NS3-4A protease inhibition. 16 17 18 Keywords: Antiviral compounds, Quinazoline derivatives, Hepatitis C virus, NS3-4A serine 19 protease, HCV replicon assay. 20 21 22 23 24 25 26 the inhibitory potential of five unique quinazoline derivatives towards HCV NS3-4A serine 1 protease and their ability to inhibit HCV replication in a cell-based HCV replicon system. 2 Methods 3 Chemical synthesis 4 Compound synthesis and characterization were carried out as previously described (17]. In 5 brief, one equivalent of amino benzhydrazide (2.5 mmol) was dissolved in 50 ml of ethanol. 6 Two equivalent of substituted aromatic salicylaldehyde (5.0 mmol) was then added to this 7 solution in the presence of glacial acetic acid (added as a catalyst at 50-60℃). The reaction 8 mixture was refluxed for 2-3h. The yellow to brown precipitates of the compounds below 9 were formed during the reactions. 10 Expression and Purification of HCV Protease 11 Protein purification was conducted as previously described with modification[18,19]. The 12 long oligonucleotides spanning DNA sequences of HCV genotype 3 proteases (NS3-GSGS-4A 13 pro) were overlapped and extended using Klenow DNA polymerase (Invitrogen, USA). PCR 14 subsequently amplifies the designed full-sized DNA and cloned into E. coli expression vector, 15 PQE30. The recombinant plasmids were then isolated, and the purified plasmids were 16 sequenced to verify the sequence identity of the insert. E. coli harboring the recombinant 17 protease vector was cultured in 1 L of LB medium and expression was induced by addition of 18 1mM IPTG followed by an additional 4 hours incubation at 370C. The bacterial cells were 19 then harvested, lysed, and sonicated on ice. Recombinant protein in the supernatan...

show abstract

Discovering novel anti-HCV compounds with inhibitory activities toward HCV NS3/4A protease

Cited by 13 publications

References 30 publications

Betulin from Hedyotis hedyotidea ameliorates concanavalin A-induced and T cell-mediated autoimmune hepatitis in mice

Betulin from Hedyotis hedyotidea ameliorates concanavalin A-induced and T cell-mediated autoimmune hepatitis in mice

Discovery of Metal Ions Chelator Quercetin Derivatives with Potent Anti-HCV Activities

Novel Quinazoline derivatives inhibited HCV Serine protease and viral replication in Huh-7 cells

Contact Info

Product

Resources

About