AASLD5050 一种新的以肝脏为中心的小分子的临床前表征 通过激

StephenW

5050
一种新的以肝脏为中心的小分子的临床前表征
通过激活 I 型干扰素信号有效抑制乙型肝炎病毒
Ariel Tang, Lida Guo, Xiang Xu, Nuruddin Unchwaniwala, Lewyn Li, Carl Li, Michael Shen, Jiaxin Yu, Hassan
Pajouhesh、Marc P. Windisch、Michel Perron、Michael A. Walker、William Delaney、Min Zhong 和 Ken Zhang，
装配生物科学
背景：慢性 HBV 感染 (cHBV) 影响全球约 2.96 亿患者。核苷（酸）ide反向
转录酶抑制剂可减少 HBV DNA，但治疗是不确定的。聚乙二醇化干扰素 α (PEG-IFN-α)，
具有免疫调节和抗病毒活性，导致部分患者的 HBsAg 清除（功能性治愈）。
然而，较差的耐受性限制了其临床应用。在这里，我们描述了一种新型口服药物的临床前概况
通过激活 IFN 信号抑制 HBV 并参与免疫系统的生物可利用小分子
途径。方法：在受感染的原代人类中测量 HBV 和丙型肝炎病毒 (HCV) 抑制的 EC50
HBeAg ELISA 检测肝细胞 (PHH)，荧光素酶报告基因检测 Huh-7 复制子细胞。 IC50s 干扰素-
在 HEK293 细胞中测量敏感反应元件 (ISRE) 报告基因的活性。干扰素刺激基因 (ISG)
通过 RT-qPCR 和 RNA 杂交 (NanoString) 在小鼠肝脏（体内）和 PHH（体外）中评估诱导，
分别。在 HCV 复制子细胞中评估了诱导抗病毒作用的化合物暴露时间。 JAK-1
使用 JAK-1 特异性抑制剂评估依赖性，并通过蛋白质印迹评估 STAT-1 磷酸化。
在小鼠中进行药代动力学（PK）和药效学（PD）实验。结果：结构上
选择不同的化合物进行效力和 PK/PD 分析。所有化合物均抑制 HBV (EC50 0.8-5
μM)、HCV (EC50 0.03-0.2 μM) 和诱导 ISRE 报告基因活性 (IC50 2-9 μM)。 JAK-1 抑制剂阻止抗病毒
效应和 STAT-1 磷酸化。在体外，只需 15 分钟的化合物暴露即可抑制 HCV
复制，而需要 24 小时的复合暴露才能达到与 IFN-α 相同的效果。这
小分子诱导 ISG 的幅度与 PHHs 和小鼠肝脏中的 IFN-α 相当。化合物
在啮齿动物中表现出良好的吸收（小鼠 F = 19-105%，大鼠为 69-124%），肝脏暴露增加（7-12 倍
大于血浆）和一系列终末半衰期（1.6-32.5 小时）。结论：我们发现了一类新的
口服生物可利用的小分子，激活 I 型 IFN 信号传导，并有效抑制 HBV 和 HCV。带领
正在进行优化，目的是提名一个以肝脏为重点的开发候选人
cHBV治疗的评估。
披露：
Michel Perron - 装配生物科学：就业； Assembly Biosciences：股东；

StephenW

5050
PRECLINICAL CHARACTERIZATION OF A NOVEL LIVER-FOCUSED SMALL MOLECULE
EFFICIENTLY INHIBITING HEPATITIS B VIRUS BY ACTIVATING TYPE I INTERFERON SIGNALING
Ariel Tang, Lida Guo, Xiang Xu, Nuruddin Unchwaniwala, Lewyn Li, Carl Li, Michael Shen, Jiaxin Yu, Hassan
Pajouhesh, Marc P. Windisch, Michel Perron, Michael A. Walker, William Delaney, Min Zhong and Ken Zhang,
Assembly Biosciences
Background: Chronic HBV infection (cHBV) affects ~296 million patients worldwide. Nucleos(t)ide reverse
transcriptase inhibitors reduce HBV DNA, but treatment is indefinite. Pegylated interferon alpha (PEG-IFN-α), which
has immunomodulatory and antiviral activities, leads to HBsAg clearance (functional cure) in a subset of patients.
However, poor tolerability limits its clinical use. Here we describe the preclinical profile of a novel class of orally
bioavailable small molecules that inhibit HBV and engage the immune system through activating IFN signaling
pathways. Methods: EC50s for HBV and hepatitis C virus (HCV) inhibition were measured in infected primary human
hepatocytes (PHHs) by HBeAg ELISA and Huh-7 replicon cells by luciferase reporter, respectively. IC50s for interferon-
sensitive response element (ISRE) reporter activity were measured in HEK293 cells. Interferon stimulated gene (ISG)
induction was assessed by RT-qPCR and RNA hybridization (NanoString) in mouse liver (in vivo) and PHHs (in vitro),
respectively. Compound exposure time to induce antiviral effects was evaluated in HCV replicon cells. JAK-1
dependency was assessed with JAK-1-specific inhibitors and STAT-1 phosphorylation assessed by Western blot.
Pharmacokinetic (PK) and pharmacodynamic (PD) experiments were performed in mice. Results: Structurally
differentiated compounds were selected for potency and PK/PD profiling. All compounds inhibited HBV (EC50 0.8-5
μM), HCV (EC50 0.03-0.2 μM), and induced ISRE reporter activity (IC50 2-9 μM). A JAK-1 inhibitor prevented antiviral
effects and STAT-1 phosphorylation. In vitro, only 15 minutes of compound exposure was required to inhibit HCV
replication, whereas 24 hours of compound exposure was required to achieve the same effect with IFN-α. The
magnitude of ISG induction by small molecules was comparable to IFN-α in PHHs and mouse liver. Compounds
showed good absorption in rodents (F=19-105% in mice and 69-124% in rats) with enhanced liver exposure (7-12-fold
greater than plasma) and a range of terminal half-lives (1.6-32.5 hr). Conclusion: We have identified a novel class of
small molecules that are orally bioavailable, activate type I IFN signaling, and efficiently inhibit HBV and HCV. Lead
optimization is ongoing with the objective of nominating a development candidate with liver-focused exposure for
evaluation for treatment of cHBV.
Disclosures:
Michel Perron - Assembly Biosciences: Employment; Assembly Biosciences: Stock Shareholder;