SYNTHESIS, HYBRIDIZATION PROPERTIES AND ANTIVIRAL ACTIVITY OF LIPID-OLIGODEOXYNUCLEOTIDE CONJUGATES
SYNTHESIS, HYBRIDIZATION PROPERTIES AND ANTIVIRAL ACTIVITY OF LIPID-OLIGODEOXYNUCLEOTIDE CONJUGATES
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DOI：
10.1093/nar/18.13.3777
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发表时间：
1990-07-11
影响因子：
14.9
通讯作者：
BISCHOFBERGER, N
中科院分区：
文献类型：
--
作者：
SHEA, RG;MARSTERS, JC;BISCHOFBERGER, N
通过磷酸氢盐固相支持DNA合成法将1，2-二-O-十六烷基-外消旋-甘油基-3-H-膦酸三乙铵（2）偶联到寡脱氧核苷酸的5“末端。具有单个5“-磷脂的双链体DNA寡聚物在比相应的未修饰的双链体更低的温度下熔化，但是在每个5”末端具有脂质的双链体具有更高的Tms。在L929细胞的摄取实验中，与未修饰的DNA相比，8-10倍的脂质DNA与细胞结合。未修饰的反义二酯在L929细胞中的VSV抗病毒测定中是无活性的（最高达200 μ M）。然而，脂质与寡聚体的连接导致病毒蛋白质合成在150 μ M时减少> 90%（在100 μ M时减少> 80%）。抗病毒活性依赖于寡脱氧核苷酸的序列，但一些化合物与病毒靶点具有很少或没有碱基互补性也是有效的。在VSV测定中，硫代磷酸酯衍生物在100 μ M时使病毒蛋白质合成减少20-30%。脂质-DNA化合物在高达100 μ M时对细胞没有毒性。
Triethylammonium 1,2-di-O-hexadecyl-rac-glycero-3-H-phophonate (2) was coupled to the 5'' terminus of oligodeoxynucleotides via hydrogen phosphonate solid support DNA synthesis methodology. Duplex DNA oligomers with a single 5''-phospholipid melted at lower temperatures than the corresponding unmodified duplex, but duplexes bearing lipids at each 5'' end had higher Tms. In uptake experiments with L929 cells, 8-10 times more lipid-DNA became cell-associated than did unmodified DNA. Unmodified antisense diesters were inactive in a VSV antiviral assay in L929 cells (at up to 200 .mu.M). Attachment of a lipid to the oligomer, however, led to a > 90% at 150 .mu.M (> 80% at 100 .mu.M) reduction in viral protein synthesis. The antiviral activity depended on the sequence of the oligodeoxynucleotide, but some compounds having little or no base complementarity to the viral target were also effective. Phosphorothioate derivatives reduced viral protein synthesis by 20-30% at 100 .mu.M in the VSV assay. The lipid-DNA compounds were not toxic to the cells at up to 100 .mu.M.