Methods for H-phosphonate synthesis of mono- and oligonucleotides
First Claim
1. A method of synthesizing a mononucleoside H-phosphonate comprising contacting a mononucleoside having a free 3'"'"' hydroxyl moiety with phosphonic acid and benzoic anhydride in a reaction mixture.
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Abstract
New methods of synthesizing mono- and oligo- nucleotide H-phosphonates are disclosed. The methods comprise contacting a mononucleoside with phosphonic acid and benzoyl anyhydride to yield the corresponding mononucleoside H-phosphonate. Preferrably a catalytic amount of triphosgene is also used. A similar procedure can be used to couple a first mononucleoside to a second mononucleoside or to an oligonucleotide, the method comprising contacting, in the presence of benzoic anhydride and, preferrably, a catalytic amount of triphosgene, a mononucleotide or oligonucleotide having a free 5'"'"' hydroxyl with a mononucleoside having a 3'"'"' hydroxyl-bearing phosphorous moiety (preferably H-phosphonate).
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Citations
18 Claims
- 1. A method of synthesizing a mononucleoside H-phosphonate comprising contacting a mononucleoside having a free 3'"'"' hydroxyl moiety with phosphonic acid and benzoic anhydride in a reaction mixture.
- 3. A method of synthesizing a dinucleotide comprising contacting a mononucleoside having hydroxyl-bearing phosphorous moiety at the 3'"'"' position with a mononucleoside having a free 5'"'"' hydroxyl group in the presence of benzoic anhydride in a reaction mixture.
- 7. A method of coupling a first mono- or oligo- nucleotide to a second mono- or oligo- nucleotide comprising contacting a first mono- or oligo- nucleotide having a hydroxyl-bearing phosphorous moiety at the 3'"'"' position with a second mono- or oligo- nucleotide having a free 5'"'"' hydroxyl group in the presence of benzoic anhydride in a reaction mixture.
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15. A method of synthesizing an oligonucleotide comprising sequentially:
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(a) contacting a nascent mono- or oligo- nucleotide having a free 5'"'"' hydroxyl moiety with a 5'"'"' protected mono- or oligo- nucleotide having a free 3'"'"' H-phosphonate moiety in a reaction mixture containing benzoic anhydride to produce a nascent oligonucleotide bearing a 5'"'"' protecting group; (b) cleaving the protecting group from the previously produced 5'"'"' protected nascent oligonucleotide to produce a nascent oligonucleotide having a free 5'"'"' hydroxyl moiety; (c) optionally contacting the previously produced nascent oligonucleotide having a free 5'"'"' hydroxyl moiety with a 5'"'"' protected mono- or oligo- nucleotide having a free 3'"'"' H-phosphonate moiety to produce a 5'"'"' protected nascent oligonucleotide; (d) optionally repeating (b) and (c) sequentially until an oligonucleotide of the desired sequence is obtained; (e) oxidizing the oligonucleotide of (d) to yield a phosphodiester or phosphorothioate. - View Dependent Claims (18)
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Specification