Improved synthesis of trinucleotide phosphoramidites and generation of randomized oligonucleotide libraries.
A new method to produce a set of 20 high quality trinucleotide phosphoramidites on a 5-10 g scale each was developed. The procedure starts with condensation reactions of P-components with N-acyl nucleosides, bearing the 3 '-hydroxyl function protected with 2-azidomethylbenzoyl, to give fully protected dinucleoside phosphates 13. Upon cleavage of dimethoxytrityl group from 13, dinucleoside phosphates 16 are initially transformed into trinucleoside diphosphates 19 and then the 2-azidomethylbenzoyl is selectively removed under neutral conditions to generate trinucleoside diphosphates 5 in excellent yield. Subsequent 3 '-phosphitylation affords target trinucleotide phosphoramidites 7. When mutagenic oligonucleotides are synthesized employing mixtures of building blocks 7 as well as following the new synthetic protocol, representative oligonucleotide libraries are generated in good yields.
Yagodkin
Andrey
A
Department of Pharmaceutical Chemistry, University of Kuopio, Kuopio, Finland and Metkinen Chemistry, Kuopio, Finland.
Azhayev
Alex
A
Roivainen
Jarkko
J
Antopolsky
Maxim
M
Kayushin
Alexei
A
Korosteleva
Maria
M
Miroshnikov
Anatoly
A
Randolph
John
J
Mackie
Hugh
H
D002851:Chromatography, High Pressure Liquid; D008956:Models, Chemical; D015394:Molecular Structure; D009841:Oligonucleotides / Q000737:chemistry*; D009943:Organophosphorus Compounds / Q000138:chemical synthesis* / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization; D013873:Thionucleotides / Q000737:chemistry
D009841:Oligonucleotides; D009943:Organophosphorus Compounds; D013873:Thionucleotides; C434331:phosphoramidite