Discovery of pentacyclic triterpene conjugates as HBV polymerase/NTCP dual-targeting inhibitors with potent anti-HBV activities

IF 4.5 2区医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Bioorganic Chemistry Pub Date : 2025-01-01 DOI:10.1016/j.bioorg.2024.108054

Yixin Chen , Meitao Duan , Jianling Xu , Ao Duan , Haocheng Yang , Hongquan Tao , Shuo Tian , Zishan Zhou , Wenzhang Li , Huaming Tao , Yongyan Zhu , Quanhong Zhu

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引用次数: 0

Abstract

The inhibition of HBV DNA and elimination of HBsAg has already been established as an indicator for HBV clinic cure, and a novel dual-targeting inhibitors of HBV polymerase/entry are designed and synthesized in this study. Pentacyclic triterpenes (PTs) scaffold of exhibiting a high affinity to NTCP, including glycyrrhitinic acid (GA), oleanolic acid (OA), ursolic acid (UA), and betulinic acid (BA) were linked neatly with the nucleoside drug zidovudine (AZT) through a molecular hybrid strategy to synthesize twenty of PTs-AZT conjugates for targeting HBV polymerase as well as sodium taurocholate cotransporting polypeptide (NTCP). The conjugates showed significant inhibitory effects on the secretion of HBsAg and HBeAg in HepG2.2.15 cells, and the activity on HBsAg were better. Moreover, HBV DNA replication was also notably suppressed after incubated with the conjugates. The IC₅₀ value of BA-AZT1 on HBsAg inhibition was 0.65 ± 0.07 μM, and it was 284.2 times and 442.2 times higher comparing to corresponding parent compound BA and AZT. Additionally, the therapeutic index (TI) was also improved by 87.8 times than AZT. And the IC₅₀ value of BA-AZT1 on inhibition of HBV DNA replication was 0.70 ± 0.02 μM, 10.4 times higher than that of AZT besides conspicuous TI. Molecular docking suggested that AZT skeleton of conjugate BA-AZT1 interacted with B region of HBV Polymerase reverse transcription region, and BA structure simultaneously targeted to C region of polymerase via hydrophobic chain, establishing strong binding interactions with the HBV Pol protein. In addition, docked with NTCP, BA-AZT1 with flat pentacyclic structure inserted into the interface and also formed hydrogen bonds, hydrophobic and van der Waals forces with the amino residue 157–165 of NTCP. Further SPR analysis demonstrated the binding affinity of BA-AZT1 to C region of polymerase was 19.55 μM, stronger than 53.21 μM of BA and 31.82 μM of AZT. BA-AZT1 selectively bound to the 157–165 epitopes of NTCP receptors in host cell but not PreS1 of virus. As a result, we deduced that the designed conjugates targeted NTCP and HBV polymerase, not only prevented HBV from entering host cells via selective binding NTCP, but also inhibited HBV DNA replication through obstructing the function of HBV polymerase, and it could potentially serve as a promising dual-functional and dual-target inhibitor with both replication and entry inhibition to exert anti-HBV activity.

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发现五环三萜缀合物作为HBV聚合酶/NTCP双靶向抑制剂具有有效的抗HBV活性。

抑制 HBV DNA 和消除 HBsAg 已被确定为 HBV 临床治愈的指标，本研究设计并合成了一种新型 HBV 聚合酶/入口双靶向抑制剂。五环三萜（PTs）支架对 NTCP 具有高亲和力，包括甘草次酸（GA）、齐墩果酸（OA）、熊果酸（UA）、通过分子杂交策略，将甘草次酸（GA）、齐墩果酸（OA）、熊果酸（UA）和白桦脂酸（BA）与核苷类药物齐多夫定（AZT）整齐地连接在一起，合成了 20 种 PTs-AZT 共轭物，用于靶向 HBV 聚合酶和牛磺胆酸钠共转运多肽（NTCP）。这些共轭物对 HepG2.2.15 细胞中 HBsAg 和 HBeAg 的分泌有明显的抑制作用，对 HBsAg 的活性更好。此外，HBV DNA 复制在与共轭物培养后也明显受到抑制。BA-AZT1 抑制 HBsAg 的 IC50 值为 0.65 ± 0.07 μM，是相应母体化合物 BA 和 AZT 的 284.2 倍和 442.2 倍。此外，治疗指数（TI）也比 AZT 提高了 87.8 倍。BA-AZT1 抑制 HBV DNA 复制的 IC50 值为 0.70 ± 0.02 μM，是 AZT 的 10.4 倍，而且治疗指数也非常明显。分子对接表明，BA-AZT1共轭物的AZT骨架与HBV聚合酶反转录区的B区相互作用，BA结构同时通过疏水链靶向聚合酶的C区，与HBV Pol蛋白建立了很强的结合相互作用。此外，BA-AZT1与NTCP对接后，其扁平的五环结构插入界面，并与NTCP的氨基残基157-165形成氢键、疏水和范德华力。进一步的 SPR 分析表明，BA-AZT1 与聚合酶 C 区的结合亲和力为 19.55 μM，强于 BA 的 53.21 μM 和 AZT 的 31.82 μM。BA-AZT1 能选择性地与宿主细胞中的 NTCP 受体的 157-165 表位结合，而不能与病毒的 PreS1 结合。因此，我们推断所设计的共轭物以 NTCP 和 HBV 聚合酶为靶点，不仅通过选择性结合 NTCP 阻止 HBV 进入宿主细胞，还通过阻碍 HBV 聚合酶的功能抑制 HBV DNA 复制，有可能成为一种有前途的双功能、双靶点抑制剂，同时具有抑制复制和进入的作用，从而发挥抗 HBV 的活性。

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来源期刊

Bioorganic Chemistry 生物-生化与分子生物学

CiteScore

9.70

自引率

3.90%

发文量

679

审稿时长

31 days

期刊介绍： Bioorganic Chemistry publishes research that addresses biological questions at the molecular level, using organic chemistry and principles of physical organic chemistry. The scope of the journal covers a range of topics at the organic chemistry-biology interface, including: enzyme catalysis, biotransformation and enzyme inhibition; nucleic acids chemistry; medicinal chemistry; natural product chemistry, natural product synthesis and natural product biosynthesis; antimicrobial agents; lipid and peptide chemistry; biophysical chemistry; biological probes; bio-orthogonal chemistry and biomimetic chemistry. For manuscripts dealing with synthetic bioactive compounds, the Journal requires that the molecular target of the compounds described must be known, and must be demonstrated experimentally in the manuscript. For studies involving natural products, if the molecular target is unknown, some data beyond simple cell-based toxicity studies to provide insight into the mechanism of action is required. Studies supported by molecular docking are welcome, but must be supported by experimental data. The Journal does not consider manuscripts that are purely theoretical or computational in nature. The Journal publishes regular articles, short communications and reviews. Reviews are normally invited by Editors or Editorial Board members. Authors of unsolicited reviews should first contact an Editor or Editorial Board member to determine whether the proposed article is within the scope of the Journal.