新型抗HBV长效4'修饰核苷逆转录酶抑制剂的鉴定

StephenW

Identification of a novel long-acting 4’-modified nucleoside reverse transcriptase inhibitor against HBV

    Nobuyo Higashi-Kuwata
    Sanae Hayashi
    Hiroki Kumamoto
    Masakazu Kakuni
    Yasuhito Tanaka
    Hiroaki Mitsuya
    Show all authors

Open AccessPublishedecember 14, 2020DOI:https://doi.org/10.1016/j.jhep.2020.12.006
Highlights

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    E-CFCP potently reduces HBV viremia without significant toxicity in human liver chimeric mice.
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    E-CFCP efficiently penetrates into and is tri-phosphorylated in human hepatocytes.
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    E-CFCP represents the first reported long-acting NRTI with potent anti-HBV activity.

Background & Aims
While certain nucleos(t)ide reverse transcriptase inhibitors (NRTIs) are efficacious in treating HBV infection, their effects are yet to be optimized and the emergence of NRTI-resistant HBV variants is an issue because of the requirement for lifelong treatment. The development of agents that more profoundly suppress wild-type and drug-resistant HBVs, and that have a long-acting effect, are crucial to improve patient outcomes.
Methods
Herein, we synthesized a novel long-acting 4’-modified NRTI termed E-CFCP. We tested its anti-HBV activity in vitro, before evaluating its anti-HBV activity in HBV-infected human-liver-chimeric mice (PXB-mice). E-CFCP’s long-acting features and E-CFCP-triphosphate’s interactions with the HBV reverse transcriptase (HBV-RT) were examined.
Results
E-CFCP potently blocked HBVWTD1 production (IC50qPCR_cell=1.8 nM) in HepG2.2.15 cells and HBVWTC2 (IC50SB_cell=0.7 nM), entecavir (ETV)-resistant HBVETV-RL180M/S202G/M204V (IC50SB_cell=77.5 nM), and adefovir-resistant HBVADV-RA181T/N236T production (IC50SB_cell=14.1 nM) in Huh7 cells. E-CFCP profoundly inhibited intracellular HBV DNA production to below the detection limit, but ETV and tenofovir alafenamide (TAF) failed to do so. E-CFCP also showed less toxicity than ETV and TAF. E-CFCP better penetrated hepatocytes and was better tri-phosphorylated; E-CFCP-triphosphate persisted intracellularly for longer than ETV-triphosphate. Once-daily peroral E-CFCP administration over 2 weeks (0.02~0.2 mg/kg/day) reduced HBVWTC2-viremia by 2–3 logs in PXB-mice without significant toxicities and the reduction persisted over 1–3 weeks following treatment cessation, suggesting once-weekly dosing capabilities. E-CFCP also reduced HBVETV-RL180M/S202G/M204V-viremia by 2 logs over 2 weeks, while ETV completely failed to reduce HBVETV-RL180M/S202G/M204V-viremia. E-CFCP’s 4’-cyano and fluorine interact with both HBVWT-RT and HBVETV-RL180M/S202G-M204 -RT via Van der Waals and polar forces, being important for E-CFCP-triphosphate’s interactions and anti-HBV potency.
Conclusion
E-CFCP represents the first reported potential long-acting NRTI with potent activity against wild-type and treatment-resistant HBV.
Lay summary
Although there are currently effective treatment options for HBV, treatment-resistant variants and the need for lifelong therapy pose a significant challenge. Therefore, the development of new treatment options is crucial to improve outcomes and quality of life. Herein, we report preclinical evidence showing that the anti-HBV agent, E-CFCP, has potent activity against wild-type and treatment-resistant variants. In addition, once-weekly oral dosing may be possible, which is preferrable to the current daily dosing regimens.

StephenW

新型抗HBV长效4'修饰核苷逆转录酶抑制剂的鉴定

    东仓信代（Nobuyo Higashi-Kuwata）
    早苗早树
    熊本弘树
    正国昌一
    田中靖人
    三谷广明（Hiroaki Mitsuya）
    显示所有作者

开放获取发布时间：2020年12月14日DOI：https：//doi.org/10.1016/j.jhep.2020.12.006
强调

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    E-CFCP可有效降低HBV病毒血症，而对人肝嵌合小鼠无明显毒性。
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    E-CFCP有效渗透到人肝细胞中并在其中被三磷酸化。
    •
    E-CFCP代表了第一个报道的具有有效抗HBV活性的长效NRTI。

背景与目标
尽管某些核苷酸（t）逆转录酶抑制剂（NRTIs）可有效治疗HBV感染，但其效果尚未得到优化，由于需要终身治疗，因此耐NRTI的HBV变异体的出现是一个问题。开发能更深刻地抑制野生型和耐药性HBV并具有长效作用的药物，对于改善患者预后至关重要。
方法
在这里，我们合成了一种新型的长效4'修饰NRTI，称为E-CFCP。在评估其在HBV感染的人肝嵌合小鼠（PXB-小鼠）中的抗HBV活性之前，我们在体外测试了其抗HBV活性。研究了E-CFCP的长效功能以及E-CFCP-三磷酸与HBV逆转录酶（HBV-RT）的相互作用。
结果
E-CFCP可以有效阻止HepG2.2.15细胞和HBVWTC2（IC50SB_cell = 0.7 nM），耐恩替卡韦（ETV）的HBVETV-RL180M / S202G / M204V（IC50SB_cell = 77.5 nM）和ade产生HBVWTD1（IC50qPCR_cell = 1.8 nM） Huh7细胞中产生耐药性HBVADV-RA181T / N236T（IC50SB_cell = 14.1 nM）。 E-CFCP可以将细胞内HBV DNA的产生抑制到检测限以下，但是ETV和Tenofovir alafenamide（TAF）却不能。 E-CFCP还显示出比ETV和TAF更低的毒性。 E-CFCP更好地穿透肝细胞，三磷酸化更好。 E-CFCP-三磷酸比ETV-三磷酸在细胞内持续更长的时间。每天口服一次E-CFCP，持续2周（0.02〜0.2 mg / kg /天），PXB小鼠的HBVWTC2-病毒血症减少2-3倍，无明显毒性，并且在停止治疗后1-3周持续减少，建议每周一次的加药能力。 E-CFCP还可以在2周内将HBVETV-RL180M / S202G / M204V-病毒血症降低2个对数，而ETV完全无法降低HBVETV-RL180M / S202G / M204V-病毒血症。 E-CFCP的4'-氰基和氟通过范德华力和极性作用力与HBVWT-RT和HBVETV-RL180M / S202G-M204 -RT相互作用，这对于E-CFCP-三磷酸的相互作用和抗HBV效能至关重要。
结论
E-CFCP代表了第一个报道的潜在的长效NRTI，具有针对野生型和耐治疗性HBV的强大活性。
放置摘要
尽管目前有针对HBV的有效治疗选择，但耐药性变异和终身治疗的需求构成了巨大的挑战。因此，开发新的治疗方案对于改善预后和生活质量至关重要。在此，我们报告了临床前证据，表明抗HBV药物E-CFCP对野生型和耐治疗性变异株具有有效的活性。另外，每周一次的口服给药是可能的，这比当前的每日给药方案更好。

StephenW

https://www.journal-of-hepatolog ... aven_jbs_etoc_email

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