Synthesis of 3′-Thioamido-Modified 3′-Deoxythymidine-5′-Triphosphates and Their Use as Chain Terminators in Sanger-Dna Sequencing

Abstract

The thioamide derivatives 3'-deoxy-5'-O-(4,4'-dimethoxytrityl)-3'-[(2-methyl-1-thioxo- propyl)amino]thymidine 1 and 3'-deoxy-5'-O-(4,4'-dimethoxytrityl)-3'-((6-([(9H-(fluo-ren-9- ylmethoxy)carbonyl]-amino)-1-thioxohexyl)amino) thymidine 2 were synthesized by regioselective thionation of their corresponding amides 7 and 8 with 2,4-bis(4-methoxyphenyl)-1,3,2,4-dithiadiphosphetane-2,4-disulfide (Lawesson's reagent). The thioamides were converted into the corresponding 5'-triphosphates 3 and 4. Compound 3 was chosen for DNA sequencing experiments and 4 was further labelled with fluorescein.

4-(Benzoylthiocarbonyl)morpholine

“…Moreover, they can be synthesized and puri®ed easily (Sosnicki et al, 2001), which is an important consideration for industrial applications. Recently, thioamides have been used as fungistatic reagents (Matysiak et al, 2000) and chain terminators in Sanger±DNA sequencing reactions (Schwarzer et al, 2001). The Willgerodt± Kindler reaction is a more convenient procedure for preparing thioamides than other routes (Kawai et al, 1999).…”

2,3-Bis(2-methylphenylimino)-1,4-dithiacyclohexane

“…More recently, some thioamides have shown a wide range of bioactivities and have been used as both anti-in¯ammatory and fungistatic reagents (Matysiak et al, 2000) and also as antituberculins. Thioamides have also served as special reagents used both as chain terminators in Sanger-DNA sequencing (Schwarzer et al, 2001) and as versatile intermediates in the syntheses of antitumor agents.…”

Copper‐Catalyzed Decarboxylative Synthesis of α‐Ketothioamides Using α,β‐Unsaturated Arylcarboxylic Acids, Alicyclic Secondary Amines and Elemental Sulfur

“…[3][4][5] Thioamide is an important functional group in organic/medicinal chemistry, [6] and a vital structural motif for the construction of biologically active molecules. α-Ketothioamides have been especially exploited for their terminating property in Sanger-DNA sequencing reaction, [7,8] and are used as in vitro PHGDH inhibitor which selectively abrogates the proliferation of cancer cells. [9] There are numerous methods for the synthesis of α-ketothioamides.…”