Silyl linker for solid-phase synthesis of nucleic acid

• US 7,786,296 B2
• Filed: 02/10/2005
• Issued: 08/31/2010
• Est. Priority Date: 02/25/2004
• Status: Expired due to Fees

• Current Assignee: Japan Science And Technology Agency
• Original Assignee: Japan Science And Technology Agency
• Inventors: Seio, Kohji, Sekine, Mitsuo, Ohkubo, Akihiro
• Primary Examiner(s): Lewis; Patrick T
• Application Number:
  US10/590,267
• Publication Number:
  US 20080051571A1
• Time in Patent Office: 2,028 Days
• Field of Search: None
• US Class Current: 536/28.1
• CPC Class Codes: C07F 7/0896 Compounds with a Si-H linkage
• CPC Class Codes: C07H 19/073 with 2-deoxyribosyl as the saccharide radical
• CPC Class Codes: C07H 19/173 with 2-deoxyribosyl as the saccharide radical
• CPC Class Codes: C07H 21/04 with deoxyribosyl as saccharide radical

Abstract

The purpose of the invention is to develop a silyl linker that can be efficiently introduced on a solid-phase support used for the synthesis of nucleic acid oligomers such as DNA. The present invention relates to a silyl linker for use in the solid-phase synthesis of nucleic acid, comprised of a compound of the general formula or its ester or salt:
H—(R1)Si(R2)-(C₆H₄)—CONH-(A)-COOH  (I)
wherein each of R1 and R2 is an alkyl or aryl group, and (A) represents a spacer moiety; a 3′-end nucleoside unit having said compound linked via an oxygen atom to the 3-position of a sugar of the nucleoside or its derivative, a solid-phase support having the 3′-end nucleoside unit, and a method for synthesis of nucleic acid oligomer with the use of said solid-phase support.

Claims

1. A silyl linker for use in the solid-phase synthesis of nucleic acid, comprised of a compound of the general formula or its ester or salt:

H—

(R1)Si(R2)-(C₆H₄)—

CONH-(A)-COOH 

 

(I)

wherein the benzene ring structure is optionally further substituted, each of R1 and R2 is an alkyl, and

(A) is an alkylene group represented by the formula —

(CH₂)_n—

wherein n is 2-18.

2. The compound according to claim 1 wherein R1 and R2 are an alkyl group having 1 to 5 carbon atoms.

3. The compound according to claim 1 wherein the benzene ring structure (—

(C₆H₄)—

) has a substituent.

4. The compound according to claim 3 wherein the substituent of the benzene ring structure is selected from the group consisting of alkyl having 1 to 4 carbon atoms, halogeno, nitro, cyano and methoxy groups.

5. A 3′

-end nucleoside unit having the compound according to claim 1 linked via an oxygen atom to the 3-position of a sugar of a nucleoside or its derivative.

6. The 3′

-end nucleoside unit according to claim 5 wherein a base constituting the nucleoside is thymine.

7. The compound according to claim 6 which is 5′

-O-(4,4′

-dimethoxytrityl)-thymidine-3′

-O-diisopropylsilyl-4-benzoylaminobutanoic acid triethylammonium.

8. A solid-phase support having the 3′

-end nucleoside unit according to claim 5 introduced thereon.

9. The solid-phase support according to claim 8 having the 3′

-end nucleoside unit at a ratio of 20-30 μ

mol/g.

10. The solid-phase support according to claim 8 or 9, which is a highly cross-linked polystyrene (HCP) solid-phase support.