Phosphorous protecting groups and methods of preparation and use thereof

Patent metadata

Field	Value
Publication number	US-11584773-B2
Application number	US-202117379288-A
Country	US
Kind code	B2
Filing date	Jul 19, 2021
Priority date	Apr 30, 2014
Publication date	Feb 21, 2023
Grant date	Feb 21, 2023

Abstract

Aspects of the present disclosure include compositions that make use of phosphorus and/or nucleobase protecting groups which find use in the synthesis of long polynucleotides. Phosphorus protecting groups are provided that help increase the stepwise coupling yield and/or phosphorous protecting groups that can be removed during the oxidation step. Amidine nucleobase protecting groups are provided that find use in the subject compositions and methods which provides for e.g., increased resistance to depurination during polynucleotide synthesis. In some instances, the methods and compositions disclosed herein utilize a combination of the phosphorus and amidine nucleobase protecting groups in the synthesis of polynucleotides having a sequence of 200 or more monomeric units in length. Also provided are methods for synthesizing a polynucleotide (e.g., a DNA) using one or more compounds disclosed herein.

First claim

What is claimed is:

1. An array of nucleic acids synthesized by a method comprising:
(a) providing a nucleoside residue having an unprotected hydroxyl group; and
(b) contacting the nucleoside residue with a nucleoside monomer to covalently bond the nucleoside monomer to the nucleoside residue, wherein the nucleoside monomer has the structural formula (I):

wherein

B is a nucleobase or an analogue thereof; wherein the B comprises an amidine nucleobase protecting group; and wherein the protecting group is described by the following structure:

wherein R 21 and R 22 are each independently an alkyl, a substituted alkyl, or R 21 and R 22 are cyclically linked to form a 5- or 6-membered substituted or unsubstituted heterocycle;
each of R 1 and R 2 is independently a linear, branched or cyclic, substituted or un-substituted alkyl, or R 1 and R 2 together form a 5-, 6-, 7- or 8-membered non-aromatic ring;
R 3 is an acid-labile protecting group; and
R is a group selected from the group consisting of a benzyl, alpha-methyl aryl, naphthalene, bi-cyclic aliphatic, S-ethylthioate and an amino acid, with the proviso that R is not o-methyl benzyl.

2. A library comprising a plurality of nucleic acids synthesized by a method comprising:
(a) providing a nucleoside residue having an unprotected hydroxyl group; and
(b) contacting the nucleoside residue with a nucleoside monomer to covalently bond the nucleoside monomer to the nucleoside residue, wherein the nucleoside monomer has the structural formula (I):

wherein

B is a nucleobase or an analogue thereof; wherein the B comprises an amidine nucleobase protecting group; and wherein the protecting group is described by the following structure:

wherein R 21 and R 22 are each independently an alkyl, a substituted alkyl, or R 21 and R 22 are cyclically linked to form a 5- or 6-membered substituted or unsubstituted heterocycle;
each of R 1 and R 2 is independently a linear, branched or cyclic, substituted or un-substituted alkyl, or R 1 and R 2 together form a 5-, 6-, 7- or 8-membered non-aromatic ring;
R 3 is an acid-labile protecting group; and
R is a group selected from the group consisting of a benzyl, alpha-methyl aryl, naphthalene, bi-cyclic aliphatic, S-ethylthioate and an amino acid, with the proviso that R is not o-methyl benzyl.

3. The array of nucleic acids of claim 1 , wherein the method further comprises exposing the nucleic acids to an oxidizing agent.

4. The array of nucleic acids of claim 1 , wherein the nucleoside residue is covalently bound to a solid support.

5. The array of nucleic acids of claim 1 , wherein at least some of the nucleic acids are a DNA having a length of about 200 to about 1,000 nucleotides.

6. The library of claim 2 , wherein the method further comprises exposing the nucleic acids to an oxidizing agent.

7. The library of claim 2 , wherein the nucleoside residue is covalently bound to a solid support.

8. The library of claim 2 , wherein at least some of the nucleic acids are a DNA having a length of about 200 to about 1,000 nucleotides.

Assignees

• Agilent Technologies Inc

Inventors

• Dellinger Douglas J
• Monfregola Luca
• Caruthers Marvin
• Roy Mithun

Classifications

• C07H21/04

  with deoxyribosyl as saccharide radical · CPC title

• C07H19/173

  with 2-deoxyribosyl as the saccharide radical · CPC title

• Y02P20/55

  Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups · CPC title

• C07H19/10 Primary

  with the saccharide radical esterified by phosphoric or polyphosphoric acids · CPC title

• C07H19/23

  Heterocyclic radicals containing two or more heterocyclic rings condensed among themselves or condensed with a common carbocyclic ring system, not provided for in groups C07H19/14 - C07H19/22 · CPC title