Protected monomer and method of final deprotection for rna synthesis
OwnerDouglas J. Dellinger, Joel Myerson +3inventors
InventorsDouglas J. Dellinger, Joel Myerson, Agnieszka Sierzchala, Geraldine F. Dellinger, Zoltan Timar
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Abstract
A nucleoside monomer that is protected by a thionocarbamate protecting group is provided, as well as a method for making a polynucleotide that uses the same. Also provided is a polynucleotide synthesis method that employs a diamine to deprotect a protected polynucleotide.
Claims40
A compound of the structure:
- wherein:
- BP is a protected or unprotected heterocycle;
- R1 and R2 are each independently selected from hydrogen, a protecting group, and a group comprising a phosphorus;
- and
- PG is a thionocarbamate protecting group.
A polynucleotide comprising:
- a ribonucleotide residue comprising the structure:
- wherein:
- BP is a protected or unprotected heterocycle;
- and
- R12 is selected from hydrogen, a hydrocarbyl, a substituted hydrocarbyl, an aryl, and a substituted aryl;
- and
- X is O or S;
- and
- PG is a thionocarbamate protecting group.
A method of deprotecting a solid support bound polynucleotide comprising at least one 2′-protected ribonucleotide residue, wherein said residue is not a 2′-ester protected ribonucleotide, said method comprising:
- contacting said polynucleotide with a composition comprising a diamine under conditions sufficient to deprotect said 2′-protected ribonucleotide residue.
A method comprising:
- (a) contacting a solid support bound polynucleotide comprising:
- a ribonucleotide residue comprising a 2′-protecting group, a phosphate protecting group, and optionally a nucleobase protecting group;
- with a first composition comprising a phosphate deprotection reagent, to remove said phosphate protecting group and produce a first deprotected polynucleotide that remains bound to said solid support;
- (b) contacting said first deprotected polynucleotide with a second composition comprising a diamine to remove said 2′-protecting group and removing the nucleobase protecting group, if present, to produce a second deprotected polynucleotide;
- and
- (c) or (d) wherein:
- (c) comprises simultaneously cleaving said second deprotected polynucleotide from said solid support;
- and
- (d) comprises contacting said second deprotected polynucleotide with a third composition comprising a linker cleaving reagent to cleave said second deprotected polynucleotide from said solid support,
- to produce a deprotected, cleaved polynucleotide.
A method of deprotecting a polynucleotide comprising a nucleobase protecting group;
- and a ribonucleotide residue comprising a 2′-protecting group selected from tert-butyldimethylsilyl (TBDMS), triisopropylsilyloxymethyl (TOM) and 2′-O-bis(2-acetoxyethoxy)methyl (ACE);
- said method comprising:
- (a) contacting said polynucleotide with a first composition comprising a 2′-deprotection reagent, under conditions sufficient to remove said 2′-protecting group and produce a first deprotected polynucleotide;
- (b) contacting said first deprotected polynucleotide with a second composition comprising a diamine, under conditions sufficient to remove said nucleobase protecting group and produce a fully deprotected polynucleotide.
A method of deprotecting a solid support bound polynucleotide comprising a phosphate protecting group, a nucleobase protecting group;
- and a ribonucleotide residue comprising 2′-protecting group;
- said method comprising:
- (a) contacting said polynucleotide with a first composition comprising a phosphate deprotection reagent, under conditions sufficient to remove said phosphate protecting group and produce a first deprotected polynucleotide;
- (b) contacting said first deprotected polynucleotide with a second composition comprising a 2′-deprotection reagent under conditions sufficient to remove said 2′-protecting group and produce a second deprotected polynucleotide;
- (c) contacting said second deprotected polynucleotide with a third composition comprising a diamine, under conditions sufficient to remove said nucleobase protecting group and produce a fully deprotected polynucleotide.
A method of deprotecting a solid support bound polynucleotide comprising a nucleobase protecting group and a ribonucleotide residue comprising a thionocarbamate protecting group, said method comprising:
- (a) contacting said polynucleotide with a composition comprising a diamine, under conditions sufficient to remove said protecting groups and cleave said polynucleotide from said solid support, and produce a cleaved polynucleotide;
- wherein said cleaved polynucleotide is retained on the solid support;
- (b) washing the solid support and said cleaved polynucleotide;
- and
- (c) eluting said cleaved polynucleotide from said solid support.
A kit for deprotecting a polynucleotide comprising a 2′ protected ribonucleotide residue, said kit comprising:
- a composition comprising 1,2-diaminoethane, or a derivative thereof.
Dependencies are inferred from the claim text.