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A novel, small anti-HBV compound reduces HBsAg and HBV-DNA by destabilizing HBV-RNA

  • Original Article―Liver, Pancreas, and Biliary Tract
  • Published:
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Abstract

Background

Currently, standard treatments for chronic hepatitis B such as nucleos(t)ide analogs (NAs), effectively reduce hepatitis B virus (HBV) loads but rarely result in a functional cure (defined as sustained HBsAg loss). We report the discovery of a novel, 4-pyridone compound, SAG-524, a potent and orally bioavailable small molecule inhibitor of HBV replication.

Methods

The antiviral characteristics and selectivity of SAG-524 and its derivative compound against HBV were evaluated in HBV-infection assays and HBV-infected chimeric urokinase-type plasminogen activator/severe combined immunodeficiency mice with humanized livers (PXB mice), alone or in combination with entecavir. Toxicity studies were conducted in mice and monkeys.

Results

SAG-524 reduced HBV-DNA (IC50 = 0.92 nM) and HBsAg (IC50 = 1.4 nM) in the supernatant of the HepG2.2.15 cells. SAG-524 selectively destabilized HBV-RNA via PAPD5, but not GAPDH or albumin mRNA, by shortening the poly(A) tail. PAPD5 may also be involved in HBV regulation via ELAVL1. In a study of HBV-infected PXB mice, SAG-524 produced potent reductions of serum HBsAg and HBcrAg, and the minimum effective dose was estimated to be 6 mg/kg/day. The combination therapy with entecavir greatly reduced HBsAg and cccDNA in the liver due to reduction of human hepatocytes with good tolerability. Administration of SAG-524 to monkeys, up to 1000 mg/kg/day for two weeks, led to no significant toxicity, as determined by blood tests and pathological images.

Conclusions

We have identified SAG-524 as novel and orally bioavailable HBV-RNA destabilizers which can reduce HBsAg and HBV-DNA levels, and possibly contribute a functional cure.

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Abbreviations

HBV:

Hepatitis B virus

CHB:

Chronic hepatitis B

NAs:

Nucleos(t)ide analogs

PEG-IFN:

Pegylated interferon

HCV:

Chronic hepatitis C virus

HIV:

Human immunodeficiency virus

BRIC assay:

5’-Bromouridine IP chase assay

BrU-RNA:

BrU-labeled RNA

PRE:

Posttranscriptional regulatory element

ARE:

AU-rich element

cccDNA:

Covalently closed circular DNA

MED:

Minimum effective dose

NTCP:

Sodium taurocholate cotransporting polypeptide

rcDNA:

Relaxed circular DNA

pgRNA:

Pregenomic RNA

dslDNA:

Double-stranded linear DNA

IC50 :

50% Inhibitory concentration

CC50 :

50% Cytotoxicity concentration

uPA/SCID mouse:

Chimeric urokinase-type plasminogen activator/severe combined immunodeficiency mouse

PXB-mouse:

Human liver chimeric uPA/SCID mouse

UTR:

Untranslated region

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Acknowledgements

Nothing.

Funding

This work was supported by a grant-in-aid from the Research Program on Hepatitis from the Japan Agency for Medical Research and Development (AMED JP23fk0310518).

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Author notes

  1. Takehisa Watanabe and Sanae Hayashi contributed equally to this work.

Authors and Affiliations

  1. Department of Gastroenterology and Hepatology, Faculty of Life Sciences, Kumamoto University, Honjo 1-1-1, Chuo, Kumamoto, 860-8556, Japan

    Takehisa Watanabe, Sanae Hayashi, Yan Zhaoyu, Hiroki Inada, Katsuya Nagaoka, Masakuni Tateyama & Yasuhito Tanaka

  2. Department of Virology and Liver Unit, Nagoya City University Graduate School of Medical Science, Nagoya, Japan

    Yasuhito Tanaka

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  1. Takehisa Watanabe
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Contributions

TW, SH, YT designed the study and analyzed the data. TW, SH, YZ, HI, KN, MT, experimented; YT conducted pharmacologic evaluation and compound synthesis. TW, SH, KN, YT wrote the article. All authors interpreted the data and reviewed and approved of the manuscript.

Corresponding author

Correspondence to Yasuhito Tanaka.

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Conflict of interest

Yasuhito Tanaka received Honoraria from AbbVie GK, Gilead Sciences, Inc, Chugai Pharmaceutical Co., Ltd., ASKA Pharmaceutical Holdings Co., Ltd., OTSUKA Pharmaceutical Co., Ltd., Takeda Pharmaceutical Co., Ltd. and GlaxoSmithKline PLC. He received Research funds from AbbVie GK, FUJIREBIO Inc, Sysmex Corp, Janssen Pharmaceutical K.K., GlaxoSmithKline PLC. and Gilead Sciences, Inc.

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Watanabe, T., Hayashi, S., Zhaoyu, Y. et al. A novel, small anti-HBV compound reduces HBsAg and HBV-DNA by destabilizing HBV-RNA. J Gastroenterol 59, 315–328 (2024). https://doi.org/10.1007/s00535-023-02070-y

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