Method for in vitro oligonucleotide synthesis using H-phosphonates
First Claim
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1. A method for preparing a polynucleotide phosphoramidate, said method comprising the steps of:
- oxidizing a carrier-bound polynucleoside H-phosphonate having more than two nucleosides with an oxidizing agent capable of oxidizing said carrier-bound polynucleoside H-phosphonate so as to yield carrier-bound polynucleoside phosphoramidate; and
decoupling said polynucleoside phosphoramidate from said carrier so as to yield said polynucleotide phosphoramidate.
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Abstract
A method is provided for the high fidelity, rapid and economical in vitro synthesis of oligonucleotides. Nucleoside H-phosphonates are condensed in seriatim using a dehydrating agent to produce a poly (nucleoside H-phosphonate). The product is oxidized to yield the desired oligonucleotide. A novel reagent is provided for multiple nucleoside additions in single cycles.
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Citations
11 Claims
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1. A method for preparing a polynucleotide phosphoramidate, said method comprising the steps of:
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oxidizing a carrier-bound polynucleoside H-phosphonate having more than two nucleosides with an oxidizing agent capable of oxidizing said carrier-bound polynucleoside H-phosphonate so as to yield carrier-bound polynucleoside phosphoramidate; and decoupling said polynucleoside phosphoramidate from said carrier so as to yield said polynucleotide phosphoramidate. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
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11. A method for preparing a polynucleotide phosphoramidate, said method comprising the steps of:
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oxidizing a carrier-bound polynucleoside H-phosphonate having more than two nucleosides in a solution of aqueous iodine and a base selected from the group consisting of NH3, MeNH2 and n-BuNH2 in a solvent selected from the group consisting of tetrahydrofuran and carbon tetrachloride so as to yield carrier-bound polynucleoside phosphoramidate; and incubating a carrier to which said polynucleoside phosphoramidate is bound in concentrated ammonium hydroxide so as to yield said polynucleotide phosphoramidate.
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Specification