Triphosphate oligonucleotide modification reagents and uses thereof

• US 7,173,125 B2
• Filed: 11/24/2003
• Issued: 02/06/2007
• Est. Priority Date: 08/01/2000
• Status: Expired due to Term

• Current Assignee: Vector Laboratories
• Original Assignee: Solulink, Incorporated (Maravai Life Sciences Holdings LLC)
• Inventors: Schwartz, David A., Hogrefe, Richard I.
• Primary Examiner(s): LEWIS, PATRICK T
• Application Number:
  US10/720,840
• Publication Number:
  US 20040214199A1
• Time in Patent Office: 1,170 Days
• Field of Search: 536/26.26, 536/22.1, 536/25.3, 536/25.32
• US Class Current: 536/26.26
• CPC Class Codes: C07H 19/00 Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
• CPC Class Codes: C07H 19/04 Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C07H 21/04 with deoxyribosyl as saccharide radical

Abstract

Hydrazino, oxyamino and carbonyl-based monomers and methods for incorporation into oligonucleotides during enzymatic synthesis are provided. Modified oligonucleotides are provided that incorporate the monomers provided herein. Immobilized oligonucleotides and oligonucleotide conjugates that contain covalent hydrazone or oxime linkages are provided. Methods for preparation of surface bound oligonucleotides are provided. Methods for the preparation of oligonucleotide conjugates are also provided.

Claims

1. A compound that has formula (II):

2. The compound of claim 1 that has any of formula:

3. The compound of claim 1 that is immobilized on a surface.

4. The compound of claim 1 that is conjugated to a second component.

5. A method for immobilizing oligonucleotides on a solid surface, comprising the step of:

reacting a compound of claim 1 or a plurality of said compounds with a solid surface;

wherein;

if the compound has a hydrazino or oxyamino group, the solid surface has a carbonyl group;

or

if the compound has a carbonyl group, the solid surface has a hydrazino or oxyamino group.

6. An immobilized oligonucleotide prepared by the method of claim 5.

7. A method for formation of an oligonucleotide conjugate, comprising the step of:

reacting the compound of claim 1 with a second component to form an oligonucleotide conjugate;

wherein the compound and the second component comprise complementary groups.

8. An oligonucleotide conjugate prepared by the method of claim 7.

9. The conjugate of claim 4, wherein the second component is selected from the group consisting of a fluorescein, a rhodamine and a cyanine dye.

10. A method of preparation of a hydrazino, oxyamino or carbonyl modified nucleoside triphosphate, comprising the steps of:

(i) derivatizing a carboxylic acid selected from an ω

-carbonyl, an ω

-protected hydrazino, and an ω

-protected oxyamino substituted carboxylic acid as the corresponding active ester;

(ii) reacting the resulting active ester with an amino substituted nucleoside triphosphate; and

(iii) deprotecting the hydrazino or oxyamino group, if present.