Heteroaryldihydropyrimidine (HAP) and Sulfamoylbenzamide (SBA) Inhibit Hepatitis B Virus Replication by Different Molecular Mechanisms

Zhou, Zheng; Hu, Tai‐Shan; Zhou, Xue; Wildum, Steffen; García-Alcalde, Fernando; Xu, Zhiheng; Wu, Daitze; Mao, Yi; Tian, Xiao‐Jun; Zhou, Yuan; Shen, Fang; Zhang, Zhisen; Tang, Guozhi; Nájera, Isabel; Yang, Guang; Shen, Hong C.; Young, John A. T.; Qin, Ning

doi:10.1038/srep42374

Cited by 114 publications

(175 citation statements)

References 40 publications

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“…Recently published results show that a compound chemically similar to 4f inhibits replication by disrupting the HBV capsid protein [22]. To determine if our compounds act by this mechanism, HBV capsid formation was monitored by negative-stain electron microscopy.…”

Section: Resultsmentioning

confidence: 99%

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Synthesis of sulfamoylbenzamide derivatives as HBV capsid assembly effector

Sari

Bouclé

Cox

et al. 2017

European Journal of Medicinal Chemistry

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The synthesis of novel series of sulfamoylbenzamides as HBV capsid assembly effector is reported. The structure was divided into five parts which were independently modified as part of our lead optimization. All synthesized compounds were evaluated for their anti-HBV activity and toxicity in human hepatocytes, lymphocytes and other cells. Additionally, we assessed their effect on HBV cccDNA formation in an HBeAg reporter cell-based assay. Among the 27 compounds reported, several analogs exhibited submicromolar activities and significant reduction of HBeAg secretion. Selected compounds were studied under negative-stain electron microscopy for their ability to disrupt the HBV capsid formation. Structures were modeled into a binding site recently identified in the HBV capsid protein for similar molecules to rationalize the structure-activity relationships for this family of compounds.

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Section: Resultsmentioning

confidence: 99%

“…A recent crystal structure revealed the binding site for sulfamoyl-based HBV capsid effectors at the Cp149 chain B-C dimer-dimer interface (Figure 4A) [22]. Chain B formed a “concave” that contributes most protein-ligand interactions while chain C acted as a “lid”.…”

Section: Resultsmentioning

confidence: 99%

Synthesis of sulfamoylbenzamide derivatives as HBV capsid assembly effector

Sari

Bouclé

Cox

et al. 2017

European Journal of Medicinal Chemistry

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“…Crystal structures showed that compounds from the HAP, phenylpropenamide, and sulfamoylbenzamide series target the same hydrophobic pocket located at the core dimer-dimer interface (9, 12–14). By binding to the core protein, CAMs accelerate assembly and interfere with pgRNA encapsidation and viral replication in vitro in HBV-replicating cell lines (9, 11, 15–17).…”

Section: Introductionmentioning

confidence: 99%

Hepatitis B Virus Capsid Assembly Modulators, but Not Nucleoside Analogs, Inhibit the Production of Extracellular Pregenomic RNA and Spliced RNA Variants

Lam

Ren

Espiritu

et al. 2017

Antimicrob Agents Chemother

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The hepatitis B virus (HBV) core protein serves multiple essential functions in the viral life cycle, and antiviral agents that target the core protein are being developed. Capsid assembly modulators (CAMs) are compounds that target core and misdirect capsid assembly, resulting in the suppression of HBV replication and virion production. Besides HBV DNA, circulating HBV RNA has been detected in patient serum and can be associated with the treatment response. Here we studied the effect of HBV CAMs on the production of extracellular HBV RNA using infected HepaRG cells and primary human hepatocytes. Representative compounds from the sulfonamide carboxamide and heteroaryldihydropyrimidine series of CAMs were evaluated and compared to nucleos(t)ide analogs as inhibitors of the viral polymerase. The results showed that CAMs blocked extracellular HBV RNA with efficiencies similar to those with which they blocked pregenomic RNA (pgRNA) encapsidation, HBV DNA replication, and Dane particle production. Nucleos(t)ide analogs inhibited viral replication and virion production but not encapsidation or production of extracellular HBV RNA. Profiling of HBV RNA from both culture supernatants and patient serum showed that extracellular viral RNA consisted of pgRNA and spliced pgRNA variants with an internal deletion(s) but still retained the sequences at both the 5′ and 3′ ends. Similar variants were detected in the supernatants of infected cells with and without nucleos(t)ide analog treatment. Overall, our data demonstrate that HBV CAMs represent direct antiviral agents with a profile differentiated from that of nucleos(t)ide analogs, including the inhibition of extracellular pgRNA and spliced pgRNA.

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“…All the docked poses found the HAP binding site (Fig. 5B, C, and D), and all have a hydrogen bond with W102, a key interaction that is present in cocrystal structures with capsid assembly modulators (33)(34)(35). BA-38017 and ENAN-34017 were able to find binding poses on wild-type and all of the mutant core proteins, but for Bay 41-4109, only binding poses on wild-type and V124A were reported, with no poses being reported for the V124F and V124W mutations, suggesting that these mutations could prevent binding of Bay 41-4109.…”

mentioning

confidence: 99%

“…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). 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).…”

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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Heteroaryldihydropyrimidine (HAP) and Sulfamoylbenzamide (SBA) Inhibit Hepatitis B Virus Replication by Different Molecular Mechanisms

Cited by 114 publications

References 40 publications

Synthesis of sulfamoylbenzamide derivatives as HBV capsid assembly effector

Synthesis of sulfamoylbenzamide derivatives as HBV capsid assembly effector

Hepatitis B Virus Capsid Assembly Modulators, but Not Nucleoside Analogs, Inhibit the Production of Extracellular Pregenomic RNA and Spliced RNA Variants

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

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