A series of 2-adamantanamines with alkyl adducts of various lengths were examined for efficacy against strains of influenza A including those having an S31N mutation in M2 proton channel that confer resistance to amantadine and rimantadine. The addition of as little as one CH₂ group to the methyl adduct of the amantadine/rimantadine analogue, 2-methyl-2-aminoadamantane, led to activity in vitro against two M2 S31N viruses A/Calif/07/2009 (H1N1) and A/PR/8/34 (H1N1) but not to a third A/WS/33 (H1N1). Solid state NMR of the transmembrane domain (TMD) with a site mutation corresponding to S31N shows evidence of drug binding. But electrophysiology using the full length S31N M2 protein in HEK cells showed no blockade. A wild type strain, A/Hong Kong/1/68 (H3N2) developed resistance to representative drugs within one passage with mutations in M2 TMD, but A/Calif/07/2009 S31N was slow (>8 passages) to develop resistance in vitro, and the resistant virus had no mutations in M2 TMD. The results indicate that 2-alkyl-2-aminoadamantane derivatives with sufficient adducts can persistently block p2009 influenza A in vitro through an alternative mechanism. The observations of an HA1 mutation, N160D, near the sialic acid binding site in both 6-resistant A/Calif/07/2009(H1N1) and the broadly resistant A/WS/33(H1N1) and of an HA1 mutation, I325S, in the 6-resistant virus at a cell-culture stable site suggest that the drugs tested here may block infection by direct binding near these critical sites for virus entry to the host cell.

中文翻译：

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对 H1N1 (2009) 甲型流感具有持久体外疗效的氨基金刚烷

检查了一系列具有不同长度烷基加合物的 2-金刚烷胺对甲型流感病毒株的功效，包括在 M2 质子通道中具有 S31N 突变的那些，这些突变赋予对金刚烷胺和金刚乙胺的抗性。只需添加一个 CH ₂金刚烷胺/金刚烷胺类似物的甲基加合物 2-甲基-2-氨基金刚烷的基团在体外对两种 M2 S31N 病毒 A/Calif/07/2009 (H1N1) 和 A/PR/8/34 (H1N1) 产生活性) 但不是第三个 A/WS/33 (H1N1)。具有对应于 S31N 的位点突变的跨膜结构域 (TMD) 的固态 NMR 显示了药物结合的证据。但是在 HEK 细胞中使用全长 S31N M2 蛋白的电生理学显示没有阻断。野生型菌株 A/Hong Kong/1/68 (H3N2) 在 M2 TMD 突变的一个传代内对代表性药物产生耐药性，但 A/Calif/07/2009 S31N 在在体外，耐药病毒在 M2 TMD 中没有突变。结果表明，具有足够加合物的 2-烷基-2-氨基金刚烷衍生物可以通过替代机制在体外持续阻断 p2009 甲型流感病毒。HA1 突变 N160D 的观察结果，靠近唾液酸结合位点6耐药 A/Calif/07/2009(H1N1) 和广泛耐药 A/WS/33(H1N1) 以及 HA1 突变 I325S，在细胞培养稳定位点的6耐药病毒中表明这些药物此处测试可能通过在病毒进入宿主细胞的这些关键位点附近直接结合来阻止感染。