Bigelow J C, Chrin L R, Mathews L A, McCormick J J
Department of Pharmacology, University of Vermont, Burlington 05405.
J Chromatogr. 1990 Nov 30;533:133-40. doi: 10.1016/s0378-4347(00)82193-3.
Phosphorothioate oligodeoxynucleotides (S-ODNs) have potential as anti-viral agents and are being investigated for the chemotherapy of AIDS. A high-performance liquid chromatographic method is described for the analysis, in urine and plasma, of a 28-unit deoxycytidine homopolymer (S-dC28) and a 28-unit S-ODN "antisense" to the rev gene of the human immunodeficiency virus. This method employs ion-pairing HPLC with a polymeric column. Tetrabutylammonium is used as the ion-pairing agent in a mobile phase of acetonitrile in pH 7.0 phosphate buffer. Analysis of the S-ODNs is relatively rapid (20 min) and sensitive (20 nm) and is accomplished by a gradient elution (22.5-30.0% acetonitrile) followed by ultraviolet (266 or 272 nm) absorption detection. This method is likely applicable, with appropriate modifications, to all S-ODNs of similar molecular weight regardless of sequence. The S-ODNs bind very strongly to plasma proteins but are readily prepared for analysis by a phenol extraction procedure. In a preliminary pharmacokinetic study in mice with S-dC28, very rapid elimination of the oligomer from plasma was observed (half-time, 11.6 min). Estimates for the apparent volume of distribution and total body clearance were 3 ml and 0.2 ml/min, respectively. It appears that the majority of the oligomer is eliminated by renal clearance (glomerular filtration), a property likely shared by all S-ODNs of similar molecular mass.
硫代磷酸酯寡脱氧核苷酸（S-ODNs）具有作为抗病毒药物的潜力，目前正被用于艾滋病化疗的研究。本文描述了一种高效液相色谱法，用于分析尿液和血浆中的一种28个单元的脱氧胞苷均聚物（S-dC28）以及一种针对人类免疫缺陷病毒rev基因的28个单元的S-ODN“反义链”。该方法采用离子对高效液相色谱法，使用聚合物柱。四丁基铵用作离子对试剂，流动相为pH 7.0磷酸盐缓冲液中的乙腈。S-ODNs的分析相对快速（20分钟）且灵敏（20纳米），通过梯度洗脱（22.5 - 30.0%乙腈）后进行紫外（266或272纳米）吸收检测来完成。该方法经过适当修改后，可能适用于所有分子量相似的S-ODNs，无论其序列如何。S-ODNs与血浆蛋白结合非常紧密，但通过酚提取程序很容易制备用于分析。在对小鼠进行的S-dC28初步药代动力学研究中，观察到该寡聚物从血浆中快速消除（半衰期为11.6分钟）。分布表观体积和全身清除率的估计值分别为3毫升和0.2毫升/分钟。看来大部分寡聚物是通过肾脏清除（肾小球滤过）消除的，这可能是所有分子量相似的S-ODNs共有的特性。
