Preclinical Characterization of GLS4, an Inhibitor of Hepatitis B Virus Core Particle Assembly

Wu, Guobin; Liu, Bo; Zhang, Yingjun; Li, Jing; Arzumanyan, Alla; Clayton, Marcia M.; Schinazi, Raymond F.; Wang, Zhaohe; Goldmann, Siegfried; Ren, Qingyun; Zhang, Faxhou; Feitelson, Mark A.

doi:10.1128/aac.01091-13

Cited by 106 publications

(85 citation statements)

References 46 publications

(59 reference statements)

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“…Recent significant progress includes the development of potent HBV entry inhibitors (Petersen et al, 2008), assembly inhibitors (Campagna et al, 2013;Wu et al, 2013;Yang et al, 2014) and covalently closed circular DNA (cccDNA) inhibitors (Cai et al, 2012), as well as efficient and safe technologies, such as the clustered regularly-interspaced short palindromic repeat (CRISPR)/ Cas9 system and IFN-mediated degradation, which can be used to specifically degrade cccDNA (Lin et al, 2014;Lucifora et al, 2014). Moreover, many researchers are committed to identifying new cellular targets involved in HBV replication, and in innate and adaptive immune responses for anti-HBV therapy, and to developing related inhibitors such as Hsp90 inhibitors, Hsc70 inhibitors and TLR7 agonists (Hu et al, 2004;Lopatin et al, 2013).…”

Section: To Date Two Ifns [Ifn-a and Pegylated Ifn (Peg-ifn-a)]mentioning

confidence: 99%

Oxethazaine inhibits hepatitis B virus capsid assembly by blocking the cytosolic calcium‐signalling pathway

Zhang

Liu

Xiao

et al. 2016

Journal of General Virology

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Chronic hepatitis B virus (HBV) infection is a serious public health problem and may progress to liver fibrosis, cirrhosis and hepatocellular carcinoma. It is currently treated with PEGylated IFN-a2a and nucleoside/nucleotide analogues (NAs). However, PEGylated IFN treatment has problems of high cost, low efficiency and side effects. Long-term administration of NAs is necessary to avoid virus relapse, which can cause drug resistance and side effects. New efforts are now being directed to develop novel anti-HBV drugs targeting either additional viral targets other than viral DNA polymerase or host targets to improve the treatment of chronic hepatitis B. In this study, we discovered that oxethazaine, approved for clinic use in a few countries such as Japan, India, South Africa and Brazil, can dose-dependently reduce the levels of HBV envelope antigen, extracellular HBV DNA in supernatants and intracellular HBV total DNA. However, the levels of HBV cccDNA and HBV RNAs were not affected by oxethazaine treatment. Further study confirmed that oxethazaine acts on the virus assembly stage of the HBV life cycle. A study of the mechanisms of oxethazaine suggested that this drug inhibits HBV replication and capsid assembly by blocking the cytosolic calcium-signalling pathway. Moreover, oxethazaine could inhibit the replication of lamivudine/entecavir-dual-resistant and adefovir-resistant HBV mutants. In conclusion, our study suggests that oxethazaine may serve as a promising drug, or could be used as a starting point for anti-HBV drug discovery.

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Section: To Date Two Ifns [Ifn-a and Pegylated Ifn (Peg-ifn-a)]mentioning

confidence: 99%

Oxethazaine inhibits hepatitis B virus capsid assembly by blocking the cytosolic calcium‐signalling pathway

Zhang

Liu

Xiao

et al. 2016

Journal of General Virology

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“…Most of these compounds have the ability to induce aggregation of the HBV core N-terminal assembly domain in vitro. Representative HBV core inhibitors from the heteroaryldihydropyrimidine (HAP) series of compounds, BAY 41-4109 and GLS4, have also shown antiviral activity in vivo, in mouse models of HBV replication (12)(13)(14). However, drug-like properties of current leads have not been optimized, and new classes of HBV core protein targeting compounds need to be developed to maximize antiviral efficacy as well as pharmacokinetic and safety profiles.…”

mentioning

confidence: 99%

High-resolution crystal structure of a hepatitis B virus replication inhibitor bound to the viral core protein

Klumpp¹,

Lam²,

Lukacs

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

134

187

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The hepatitis B virus (HBV) core protein is essential for HBV replication and an important target for antiviral drug discovery. We report the first, to our knowledge, high-resolution crystal structure of an antiviral compound bound to the HBV core protein. The compound NVR-010-001-E2 can induce assembly of the HBV core wild-type and Y132A mutant proteins and thermostabilize the proteins with a T m increase of more than 10°C. NVR-010-001-E2 binds at the dimer-dimer interface of the core proteins, forms a new interaction surface promoting protein-protein interaction, induces protein assembly, and increases stability. The impact of naturally occurring core protein mutations on antiviral activity correlates with NVR-010-001-E2 binding interactions determined by crystallography. The crystal structure provides understanding of a drug efficacy mechanism related to the induction and stabilization of proteinprotein interactions and enables structure-guided design to improve antiviral potency and drug-like properties.HBV treatment | HBV inhibitor | core | capsid | protein-protein interaction

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“…While heteroaryldihydropyrimidines (HAPs), such as Bay 41-4109 and GLS4, misdirect capsid assembly to form noncapsid polymers of core proteins (25)(26)(27)(28), all other reported chemotypes of core protein assembly modulators induce the formation of various sizes of capsids devoid of viral pgRNA and DNA polymerase (29)(30)(31)(32). Structural biology studies suggest that HAPs, phenylpropenamides (PPAs), and sulfamoylbenzamides (SBAs) all bind to a hydrophobic pocket formed at the dimer-dimer interface near the C termini of core protein subunits, with contributions from two neighboring core protein dimers (33).…”

mentioning

confidence: 99%

“…Binding of these molecules in the HAP pocket induces large-scale allosteric conformational changes in core protein subunits and results in quaternary and/or tertiary structure changes of capsids (34,35). Thus far, several core protein assembly modulators have been shown to inhibit HBV replication in mouse models in vivo and are currently under preclinical or clinical development (28,36).…”

mentioning

confidence: 99%

Discovery and Mechanistic Study of Benzamide Derivatives That Modulate Hepatitis B Virus Capsid Assembly

Zhao

Zhang

et al. 2017

J Virol

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Chronic hepatitis B virus (HBV) infection is a global public health problem. Although the currently approved medications can reliably reduce the viral load and prevent the progression of liver diseases, they fail to cure the viral infection. In an effort toward discovery of novel antiviral agents against HBV, a group of benzamide (BA) derivatives that significantly reduced the amount of cytoplasmic HBV DNA were discovered. The initial lead optimization efforts identified two BA derivatives with improved antiviral activity for further mechanistic studies. Interestingly, similar to our previously reported sulfamoylbenzamides (SBAs), the BAs promote the formation of empty capsids through specific interaction with HBV core protein but not other viral and host cellular components. Genetic evidence suggested that both SBAs and BAs inhibited HBV nucleocapsid assembly by binding to the heteroaryldihydropyrimidine (HAP) pocket between core protein dimer-dimer interfaces. However, unlike SBAs, BA compounds uniquely induced the formation of empty capsids that migrated more slowly in native agarose gel electrophoresis from A36V mutant than from the wild-type core protein.Moreover, we showed that the assembly of chimeric capsids from wild-type and drug-resistant core proteins was susceptible to multiple capsid assembly modulators. Hence, HBV core protein is a dominant antiviral target that may suppress the selection of drug-resistant viruses during core protein-targeting antiviral therapy. Our studies thus indicate that BAs are a chemically and mechanistically unique type of HBV capsid assembly modulators and warranted for further development as antiviral agents against HBV.IMPORTANCE HBV core protein plays essential roles in many steps of the viral replication cycle. In addition to packaging viral pregenomic RNA (pgRNA) and DNA polymerase complex into nucleocapsids for reverse transcriptional DNA replication to take place, the core protein dimers, existing in several different quaternary structures in infected hepatocytes, participate in and regulate HBV virion assembly, capsid uncoating, and covalently closed circular DNA (cccDNA) formation. It is anticipated that small molecular core protein assembly modulators may disrupt one or multiple steps of HBV replication, depending on their interaction with the distinct quaternary structures of core protein. The discovery of novel core protein-targeting antivirals, such as benzamide derivatives reported here, and investigation of their antiviral mechanism may lead to the identification of antiviral therapeutics for the cure of chronic hepatitis B.KEYWORDS antiviral agents, capsid assembly, hepatitis B virus

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Preclinical Characterization of GLS4, an Inhibitor of Hepatitis B Virus Core Particle Assembly

Cited by 106 publications

References 46 publications

Oxethazaine inhibits hepatitis B virus capsid assembly by blocking the cytosolic calcium‐signalling pathway

Oxethazaine inhibits hepatitis B virus capsid assembly by blocking the cytosolic calcium‐signalling pathway

High-resolution crystal structure of a hepatitis B virus replication inhibitor bound to the viral core protein

Discovery and Mechanistic Study of Benzamide Derivatives That Modulate Hepatitis B Virus Capsid Assembly

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