Department of Neurochemistry, Palladin Institute of Biochemistry, Leontovich Str., 9, Kiev, 01601, Ukraine,
J Membr Biol. 2014 Mar;247(3):211-29. doi: 10.1007/s00232-013-9626-8. Epub 2014 Jan 9.
The action of antifungal drug, amphotericin B (AmB), on solvent-containing planar lipid bilayers made of sterols (cholesterol, ergosterol) and synthetic C14-C18 tail phospholipids (PCs) or egg PC has been investigated in a voltage-clamp mode. Within the range of PCs tested, a similar increase was achieved in the lifetime of one-sided AmB channels in cholesterol- and ergosterol-containing membranes with the C16 tail PC, DPhPC at sterol/DPhPC molar ratio ≤1. The AmB channel lifetimes decreased only at sterol/DPhPC molar ratio >1 that occurred with sterol/PC molar ratio of target cell membranes at a pathological state. These data obtained on bilayer membranes two times thicker than one-sided AmB channel length are consistent with the accepted AmB pore-forming mechanism, which is associated with membrane thinning around AmB-sterol complex in the lipid rafts. Our results show that AmB can create cytotoxic (long open) channels in cholesterol membrane with C14-C16 tail PCs and nontoxic (short open) channels with C17-C18 tail PCs as the lifetime of one-sided AmB channel depends on ~2-5 Å difference in the thickness of sterol-containing C16 and C18 tail PC membranes. The reduction in toxic AmB channels efficacy can be required at the drug administration because C16 tails in native membrane PCs occur almost as often as C18 tails. The comparative analysis of AmB channel blocking by tetraethylammonium chloride, tetramethylammonium chloride and thiazole derivative of vitamin B1, 3-decyloxycarbonylmethyl-4-methyl-5-(2-hydroxyethyl) thiazole chloride (DMHT), has proved that DMHT is a comparable substitute for both tetraalkylammonia that exhibits a much higher affinity.
抗真菌药物两性霉素 B(AmB)在含有固醇(胆固醇、麦角固醇)和合成 C14-C18 尾磷脂(PCs)或卵磷酯(PC)的溶剂含平面脂质双层中的作用,已在电压钳模式下进行了研究。在所测试的 PCs 范围内,在胆固醇和麦角固醇存在的膜中,与 C16 尾 PC、DPhPC 的摩尔比为≤1 时,C16 尾 PC 可使单侧面 AmB 通道的寿命相似地增加。仅当甾醇/DPhPC 的摩尔比>1 时,AmB 通道的寿命才会降低,这种情况发生在病理状态下靶细胞膜的甾醇/PC 摩尔比。这些在比单侧面 AmB 通道长度厚两倍的双层膜上获得的数据与公认的 AmB 孔形成机制一致,该机制与脂质筏中 AmB-甾醇复合物周围的膜变薄有关。我们的结果表明,AmB 可以在含有 C14-C16 尾 PC 的胆固醇膜中形成细胞毒性(长开放)通道,在含有 C17-C18 尾 PC 的膜中形成非毒性(短开放)通道,因为单侧面 AmB 通道的寿命取决于甾醇含量为 C16 和 C18 尾 PC 膜的厚度差异约为 2-5Å。由于天然膜 PCs 中的 C16 尾与 C18 尾几乎一样常见,因此可能需要减少毒性 AmB 通道的药效。通过比较四乙基氯化铵、四甲基氯化铵和维生素 B1 的噻唑衍生物 3-癸氧基羰基甲基-4-甲基-5-(2-羟乙基)噻唑氯(DMHT)对 AmB 通道的阻断作用,证明 DMHT 是两者的相当替代品,均表现出更高的亲和力。