Nucleotides. Part LXXVI : Synthesis and Properties of New Isoalloxazine (=Benzo[g]pteridine-2,4(1H,3H)-dione) Derivatives as Labels for Oligonucleotides
✍ Scribed by Evgeny Kvassiouk; Ramamurthy Charubala; Wolfgang Pfleiderer
- Publisher
- John Wiley and Sons
- Year
- 2010
- Tongue
- German
- Weight
- 233 KB
- Volume
- 93
- Category
- Article
- ISSN
- 0018-019X
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✦ Synopsis
Abstract
A series of new fluorescing 8‐(6‐hydroxyhexyl)isoalloxazine (=8‐(6‐hydroxyhexyl)benzo[g]pteridine‐2,4(1__H__,3__H__)‐dione) derivatives 4–13 were synthesized from 6‐[(6‐hydroxyhexyl)amino]uracil (2) with 1‐chloro‐4‐nitrosobenzene via 8‐chloro‐10‐(6‐hydroxyhexyl)isoalloxazine (3) and subsequent substitution of the Cl‐atom of 3 by various amines (Scheme). Analogously, 8‐substituted 10‐{3‐[(2,2‐dimethyl‐1,3‐dioxolan‐4‐yl)methoxy]propyl}isoalloxazines 19, 20, and 23–25 were prepared which yielded on deprotection the corresponding 10‐[3‐(2,3‐dihydroxypropoxy)propyl]alloxazines 21, 22, and 26–28. Their conversion into the 3″‐O‐(4,4′‐dimethoxytrityl) derivatives 29–33 and subsequent transformation into the corresponding 2″‐(2‐cyanoethyl N,N‐diisopropylphosphoramidites) 34–38 led to new building blocks for oligonucleotide synthesis. A series of 21‐mer oligodeoxyribonucleotides carrying the fluorescing isoalloxazine 37 in various positions of the chain were assembled in a DNA synthesizer. Combination with the complementary sequence yielded the stable duplexes 40–54 showing by the melting temperatures T~m~ that the fluorophor (F) does not harm the stability of the unmodified duplex 39 (Table).