Repurposing of approved drugs that target host functions also important for virus replication promises to overcome the shortage of antiviral therapeutics. Mostly, virus biology including initial screening of antivirals is studied in conventional monolayer cells. The biology of these cells differs considerably from infected tissues. 3D culture models with characteristics of human tissues may reflect more realistically the in vivo events during infection. We screened first, second, and third generation epidermal growth factor receptor (EGFR)-inhibitors with different modes of action and the EGFR-blocking monoclonal antibody cetuximab in a 3D cell culture infection model with primary human keratinocytes and cowpox virus (CPXV) for antiviral activity. Antiviral activity of erlotinib and osimertinib was nearly unaffected by the cultivation method similar to the virus-directed antivirals tecovirimat and cidofovir. In contrast, the host-directed inhibitors afatinib and cetuximab were approx. 100-fold more efficient against CPXV in the 3D infection model, similar to previous results with gefitinib. In summary, inhibition of EGFR-signaling downregulates virus replication comparable to established virus-directed antivirals. However, in contrast to virus-directed inhibitors, in vitro efficacy of host-directed antivirals might be seriously affected by cell cultivation. Results obtained for afatinib and cetuximab suggest that screening of such drugs in standard monolayer culture might underestimate their potential as antivirals.

中文翻译：

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新型抗病毒物质在3D和单层细胞培养中的差异功效

重新利用针对宿主功能的批准药物对病毒复制也很重要，有望克服抗病毒疗法的不足。通常，在常规单层细胞中研究了病毒生物学，包括初步筛选抗病毒药物。这些细胞的生物学与受感染的组织有很大的不同。具有人体组织特征的3D培养模型可以更真实地反映感染期间的体内事件。我们在具有主要人角质形成细胞和牛痘病毒（CPXV）的3D细胞培养感染模型中，筛选了具有不同作用方式的第一代，第二代和第三代表皮生长因子受体（EGFR）抑制剂和EGFR阻断性单克隆抗体西妥昔单抗。活动。厄洛替尼和奥西替尼的抗病毒活性几乎不受培养方法的影响，类似于病毒导向的抗病毒药物tecovirimat和西多福韦。相比之下，宿主导向的抑制剂afatinib和cetuximab约为。在3D感染模型中抗CPXV的效率提高了100倍，这与吉非替尼以前的结果相似。总之，与已确立的病毒定向抗病毒药相比，EGFR信号转导的抑制下调了病毒复制。但是，与病毒导向的抑制剂相反，宿主导向的抗病毒药物的体外功效可能会受到细胞培养的严重影响。阿法替尼和西妥昔单抗获得的结果表明，在标准单层培养物中筛选此类药物可能会低估其作为抗病毒药的潜力。宿主导向的抑制剂afatinib和cetuximab约为。在3D感染模型中抗CPXV的效率提高了100倍，这与吉非替尼以前的结果相似。总之，与已确立的病毒定向抗病毒药相比，EGFR信号转导的抑制下调了病毒复制。但是，与病毒导向的抑制剂相反，宿主导向的抗病毒药物的体外功效可能会受到细胞培养的严重影响。阿法替尼和西妥昔单抗获得的结果表明，在标准单层培养物中筛选此类药物可能会低估其作为抗病毒药的潜力。宿主导向的抑制剂afatinib和cetuximab约为。在3D感染模型中抗CPXV的效率提高了100倍，这与吉非替尼以前的结果相似。总之，与已确立的病毒定向抗病毒药相比，EGFR信号转导的抑制下调了病毒复制。但是，与病毒导向的抑制剂相反，宿主导向的抗病毒药物的体外功效可能会受到细胞培养的严重影响。阿法替尼和西妥昔单抗获得的结果表明，在标准单层培养物中筛选此类药物可能会低估其作为抗病毒药的潜力。抑制EGFR信号传导可下调病毒复制，其作用与已确立的针对病毒的抗病毒药相当。但是，与病毒导向的抑制剂相反，宿主导向的抗病毒药物的体外功效可能会受到细胞培养的严重影响。阿法替尼和西妥昔单抗获得的结果表明，在标准单层培养物中筛选此类药物可能会低估其作为抗病毒药的潜力。抑制EGFR信号传导可下调病毒复制，其作用与已确立的针对病毒的抗病毒药相当。但是，与病毒导向的抑制剂相反，宿主导向的抗病毒药物的体外功效可能会受到细胞培养的严重影响。阿法替尼和西妥昔单抗获得的结果表明，在标准单层培养物中筛选此类药物可能会低估其作为抗病毒药的潜力。