Post-synthetic introduction of labile functionalities onto purine residues via 6-methylthiopurines in oligodeoxyribonucleotides

Tetrahedron

Volumn 52, Issue 32, 1996, Pages 10737-10750

  Xu, Yao Zhong ^a  

^a UNIVERSITY COLLEGE LONDON   (United Kingdom)

Author keywords

[No Author keywords available]

Indexed keywords

OLIGODEOXYRIBONUCLEOTIDE; PURINE DERIVATIVE;

ARTICLE; DRUG SYNTHESIS; PRIORITY JOURNAL; REACTION ANALYSIS; REVERSED PHASE HIGH PERFORMANCE LIQUID CHROMATOGRAPHY; TECHNIQUE; ULTRAVIOLET SPECTROPHOTOMETRY;

EID: 0030570846     PISSN: 00404020     EISSN: None     Source Type: Journal    
DOI: 10.1016/0040-4020(96)00596-0     Document Type: Article

References (33)

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21. 6-ethanoadenine. Although the methylthio group was assumed to be converted into its sulphonyl group, however no direct evidence was provided.
22. 3).
23. 2) is virtually the same as that of the non-substituted pentamer (i.e. CGXAT, X: 6-methylthiopurine) obtained by direct purification without the oxidation and conversion.
24. 6-methyladenine). A prolonged substitution with methylamine did not increase the yield of the product. When the oxidizing time was increased from 5 min to 1 hr, most of the starting pentamer was transformed into the desired pentamer. It was unexpected that the oxidation of the fully deprotected pentamer was slower than that of the pentamer undeprotected and attached to the CPG support. However, it was reasoned that the slower oxidation might be due to the poor solubility of MCPBA in the aqueous buffer. As MCPBA is a rather strong oxidizing agent and could cause adverse effects on the oligomer in a prolonged treatment, therefore a mild oxidizing reagent MMPP was used instead of MCPBA for oxidation of oligomers in the aqueous solution.
25. 16. A fully deprotected pentamer containing 6-methylthiopurine, after oxidized with MMPP, desalted and dried, was dissolved in 0.1 M potassium dihydrogen phosphate (pH 6.3). The MMPP-treated oligomer solution was mixed with 0.1 M aqueous sodium sulphide (pH 12) at RT. After 1 hr, HPLC showed that the retention time of the oligomer has changed from 19 min (Peak b in Fig. 5) to 17.5 min (Peak a) and it could also be detected at the wavelength of 325 nm. When the MMPP-treated oligomer solution was mixed with 0.1 M aqueous sodium azide, HPLC showed that the new peak (Rt = 20 min, Peak c in Fig. 5) eluted later than the starting oligomer (Rt=19 min) and the hydrolysed product (i.e. the pentamer containing hypoxanthine, Rt=18 min). These observations suggest that the later-eluting (Peak c) is very likely to be the pentamer containing 6-azido-purine.
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27. 2, ex) and 8.14 (1H, s, 8-H). UV λmax=315 nm, λmin=274 nm. The preparation of 2′-deoxy-6-methylthiopurine nucleoside (4) has been described in the experimental section.
28. 3CN (v/v) was added and vigorously mixed for 5 min. The solution was left overnight and the desired oligomer was isolated using a Sep-Pac cartridge (Waters).
29. 20. The DNA duplex consisting of this sequence has been shown to have a higher yield of crosslinkage with Cro-protein [Zheng, Q. Ph.D thesis (University of London) 1995].
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