Uses and synthesis of 5-Methoxyuridine
5-Methoxyuridine is an intermediate in the biosynthesis of pyrimidines and is also used as a precursor for other compounds.
Uses of 5-methoxyuridine
5-Methoxyuridine is an analog of Uridine and is used as a reagent in the synthesis of 5-OMe-UDP, a potent and selective P2Y6-receptor agonist. 5-Methoxyuridine inhibits messenger RNA (mRNA) production in bacteria by interfering with sequence recognition and binding sites within the ribosome and preventing.
Chemical synthesis of 5-methoxyuridine
5-Hydroxyuridine was synthesized by the method described by T. Ueda10. 5-Hydroxyuridine (30 mg) was further methylated by 20 y1 of dimethyl sulfate in 0.6 ml of 0.1 N NaOH. A reaction mixture was chromatographed on Whatman 3MM paper using solvent D. The band containing mo5U was extracted with water and the extract was further purified by paper chromatography in sol vent E. These procedures gave a 25% yield of mo5l). In addition to mo5U and the raw material, the methylated products contained 5% of 5-hydroxy-3-methyl uridine and 4% of 5-methoxy-3-methyluridine.1
Related research
We could not detect mo5Up in RNase T2 hydrolyzate of whole tRNA mole cule, because mo5Up was chromatographed at the same position as Up on a thin-layer plate for base analysis. The separation of tno5U from U is dif ficult in paper or thin-layer chromatography with usual solvent systems. In case of RNase T2 hydrolyzate of C-U-U-mo5U-Gp from tRNAAla, we found the maximum UV absorption, at pH 2, of a spot corresponding to Up not at 262 nm but at 265 nm. Therefore, we suspected the presence of a new modified nucleotide. Digestion of this oligonucleotide with silkworm endonuclease resulted two fragments, i.e. C-U and pU-mo5U-Gp. RNase T2 digest of the latter fragment gave pUp, mo5Up, and Gp, then U and mo5U can be separated easily as a nucleoside diphosphate and a nucleoside monophosphate, respectively.
A sharp distinction between mo5Up and Up was established by meas urement of UV absorption spectra. If these analyses were executed with [32P]-labeled tRNA samples using radioactivity as a sole detective device, mo5U would be identified as Up and never be detected. Since the nucleoside next to the 5'-end of the anticodon is usually U, and mo5U is located the first position of the anticodon, mo5U will be contained in an RNase Ti fragment almost always together with U. These two components can be sepa rated in chromatography with usual solvent systems only when they hold dif ferent number of phosphate, namely one is nucleoside and the other is nucleotide, or one is nucleoside monophosphate and the other is nucleoside diphosphate.
