Leonard N J, McDonald J J, Reichmann M E
Proc Natl Acad Sci U S A. 1970 Sep;67(1):93-8. doi: 10.1073/pnas.67.1.93.
The use of diethyl pyrocarbonate as a nuclease inhibitor in the preparation of RNA of high molecular weight has prompted a study of the possible reactions of this compound with nucleic acid components under the conditions generally employed for providing inhibition. The first substrate investigated was adenine, which has been found to undergo ring opening with the formation of 5(4)-N-carbethoxyaminoimidazole-4(5)-N'- carbethoxycarboxamidine (II). This product was converted efficiently to isoguanine by treatment with ammonia. The structure of II was established by spectroscopy. For comparisons of reactivity and of spectroscopic and chromatographic properties with the adenine-diethyl pyrocarbonate product, the compounds 9-carbethoxyadenine, 6-N-carbethoxyaminopurine (V), and 6-ethylaminopurine were made; compound V was made by employing the 1-ethoxyethyl protecting group in the synthetic sequence. Purine compounds can be converted to 9-(1-ethoxyethyl) derivatives simply by refluxing in acetal. The facile reaction of adenine with diethyl pyrocarbonate illustrates the importance of gaining information as to the fate of various nucleic acid components in the presence of diethyl pyrocarbonate.
焦碳酸二乙酯作为一种核酸酶抑制剂用于制备高分子量RNA，这促使人们研究该化合物在通常用于提供抑制作用的条件下与核酸成分可能发生的反应。研究的第一个底物是腺嘌呤，已发现它会发生开环反应，生成5(4)-N-乙氧羰基氨基咪唑-4(5)-N'-乙氧羰基羧脒（II）。通过用氨处理，该产物能有效地转化为异鸟嘌呤。II的结构通过光谱学确定。为了将反应活性以及光谱和色谱性质与腺嘌呤 - 焦碳酸二乙酯产物进行比较，制备了化合物9-乙氧羰基腺嘌呤、6-N-乙氧羰基氨基嘌呤（V）和6-乙氨基嘌呤；化合物V是通过在合成序列中使用1-乙氧基乙基保护基团制备的。嘌呤化合物只需在缩醛中回流就能转化为9-(1-乙氧基乙基)衍生物。腺嘌呤与焦碳酸二乙酯的这种容易发生的反应说明了了解焦碳酸二乙酯存在时各种核酸成分的去向信息的重要性。
