Substituent position dictates the intercalative DNA-binding mode for anthracene-9,10-dione antitumor drugs.
Substituent position dictates the intercalative DNA-binding mode for anthracene-9,10-dione antitumor drugs.
复制标题
取代基位置决定了蒽-9,10-二酮抗肿瘤药物的插入 DNA 结合模式。
DOI：
10.1021/bi00161a050
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发表时间：
1992
期刊：
影响因子：
2.9
通讯作者：
Wilson,WD
中科院分区：
文献类型：
--
作者：
Tanious,FA;Jenkins,TC;Neidle,S;Wilson,WD
材料和方法材料。研究了两组碱性蒽-9，10-二酮类化合物：I组化合物1-4（图1），其不同之处在于被碱性部分取代的环位置，即1-单-和1，4-，1，5-和1，8-双[2-（2-甲氧基苯基）-1，4-，1，5-和1，（二乙基氨基）乙基氨基]蒽-9，10-二酮和第II族化合物5-8，以及不同碱性的2，6-取代的酰胺基蒽-9，10-二酮。先前已经报道了这些化合物的合成和纯化（Islam等人，1983，1985; Agbandje等人，1992年）。如前所述制备小牛胸腺DNA（CT-DNA，Worthington）、聚[dG-dC] 2和聚[dA-dT] 2（PL Biochemicals）（Wilson等人，1985年b，1986年）。水性MES缓冲液含有IX 10™ 2 M 2-（7 N-吗啉代）乙磺酸（MES）和IX 10™ 3 M EDTA。根据需要，加入氯化钠以将离子强度调节至所需值，并将pH调节至6.2。十二烷基硫酸钠（SDS）得自Boehringer曼海姆GmbH。用Hi-Tech SF-51停流分光光度计进行动力学测量。随仪器提供的软件用于数据采集和分析。通过与分光光度计连接的HP-330计算机中的12位高速模数转换器进行数据采集。收集吸光度与时间的单波长动力学记录。典型地，几个单独的动力学实验被计算机收集和平均，以提高信噪比。通过将等体积（50 µ）的DNA-药物复合物溶液与相同盐浓度的1%（w/w）SDS溶液混合来监测解离反应。在假一级反应条件下，用过量DNA进行了缔合动力学测定。将等体积（50 μ）的DNA溶液和化合物溶液混合。温度通过Haake A81冷冻水浴循环水控制，并通过位于SF-51样品室中的内部热敏电阻进行监测。
MATERIALS AND METHODSMaterials. Two groups of basic anthracene-9, 10-diones compounds were studied: group I compounds 1-4 (Figure 1), which differ in the positions of ring substitution by basic moieties, ie, 1-mono-and 1, 4-, 1, 5-, and l, 8-bis [2-(diethyl-amino) ethylamino] anthracene-9, 10-diones, and group II com-pounds 5-8, with varying basic 2, 6-substituted amidoanthracene-9, 10-diones. The synthesis and purification of these compounds have been reported previously (Islam et al., 1983, 1985; Agbandje et al., 1992). Calf thymus DNA (CT-DNA, Worthington), poly [dG-dC] 2 and poly [dA-dT] 2 (PL Biochemicals) were prepared as previously described (Wilson et al., 1985b, 1986). Aqueous MES buffer contained 1 X 10™ 2 M 2-(7V-morpholino) ethanesulfonic acid (MES) and 1 X 10™ 3 M EDTA. Sodium chloride was added to adjust the ionic strength to the desired value, as required, and the pH was adjusted to 6.2. Sodium dodecyl sulfate (SDS) was obtained from Boehringer Mannheim GmbH.Methods. Kinetics measurements were conducted with a Hi-Tech SF-51 stopped-flow spectrophotometer. The software provided with the instrument was used for both data acquisition and analysis. Data acquisition was carried out via a 12-bit high-speed analog-to-digital converter in a HP-330 computer interfaced to the spectrophotometer. Single-wavelength ki-netic records of absorbance versus time were collected. Typically, several individual kineticsexperiments were col-lected and averaged by the computer to improve the signal-to-noise ratio. Dissociation reactions were monitored by mixing equal volumes (50 µ) of a solution of the DNA-drug complex with a 1%(w/w) solution of SDS at the same salt concentration. Association kinetic measurements were con-ducted under pseudo-first-order conditions by using excess DNA. Equal volumes (50 µ) of a DNA solution and a solution of compound were mixed. The temperature was controlled by circulating water with a Haake A81 refrigerated water bath and was monitored with an internal thermistor positioned in the SF-51 sample compartment.