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US 2019/0315794 A1Published application
Compositions and Methods for Reverse Automated Nucleic Acid Synthesis
AssigneeThe University of Toledo
InventorsMatthew Ellis, Amanda Bryant-Friedrich
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Abstract
Methods for reverse automated nucleic acid synthesis, and 5′-H-phosphonates suitable for use in the same, as well as methods for making 5′-H-phosphonates, are described.
Claims26
A composition comprising Formula I:
- wherein:
- X is a trialkylammonium having a formula of NHR3, wherein R3 is alkyl;
- Z is oxygen, nitrogen, or fluorine;
- R1 is a protecting group;
- R2 is tert butyl dimethyl silyl (TBDMS), triisopropylsilyl oxymethylene, fluorenylmethyloxycarbonyl (Fmoc), alkyl, aryl, or acetyl;
- and
- B is hydrogen, a nucleobase, aryl, or a heterocycle optionally protected;
- and stereoisomers, racemates, hydrates, solvates, polymorphs, and prodrugs thereof.
5 more elements
A method for synthesizing an oligonucleotide, the method comprising:
- linking a protected first nucleoside through a first linker to a support to form a support-bound nucleoside;
- treating the support with a halogenated carboxylic acid in dichloromethane to free a 3′ hydroxyl of the support-bound nucleoside;
- coupling a second nucleoside to the first nucleoside to form a first H-phosphonate linkage, wherein the second nucleoside comprises a 5′-H-phosphonate prior to the coupling;
- capping unreacted 3′ ends by esterification to incorporate capping groups thereon;
- oxidizing the first H-phosphonate linkage to produce an oligonucleotide;
- and
- optionally, repeating the deprotection, coupling, capping, and oxidizing steps to produce an oligonucleotide having a desired sequence.
4 more elements
A method for making a 5′-H-phosphonate, the method comprising:
- protecting a nucleoside at a 2′ position with a first protecting group to form a 2′-protected nucleoside;
- protecting the 2′-protected nucleoside at a 5′ position to form a fully protected nucleoside;
- hydrolyzing the fully protected nucleoside in acid with heat to form a hydrolyzed protected nucleoside;
- silylating the hydrolyzed protected nucleoside at the 2′ position to form a silylated nucleoside;
- protecting the silylated nucleoside with a second protecting group to form a protected silylated nucleoside;
- deprotecting the protected silylated nucleoside to form a 5′ hydroxyl nucleoside;
- and
- phosphonating the 5′ hydroxyl nucleoside to form a 5′-H-phosphonate.
5 more elements
Dependencies are inferred from the claim text.