Preparation of Nucleotide Oligomer

• US 20100273999A1
• Filed: 05/07/2010
• Published: 10/28/2010
• Est. Priority Date: 11/13/2007
• Status: Active Grant

• Current Assignee: ST Pharm Co. Ltd.
• Original Assignee: Samchully Pharm. Co., Ltd. (Dong-A Socio Holdings Co., Ltd.)
• Inventors: Kim, Kyung-Il, Kayushin, Alexei, Ji, Yumi, Kim, Sungwon, Jung, Kyeong-Eun, Kim, Moon Hee
• Granted Patent: US 8,901,289 B2
• Time in Patent Office: Days
• US Class Current: 536/25.340
• CPC Class Codes: C07H 1/00 Processes for the preparation of sugar derivatives
• CPC Class Codes: C07H 19/067 with ribosyl as the saccharide radical
• CPC Class Codes: C07H 19/10 with the saccharide radical esterified by phosphoric or polyphosphoric acids
• CPC Class Codes: C07H 19/167 with ribosyl as the saccharide radical
• CPC Class Codes: C07H 19/20 with the saccharide radical esterified by phosphoric or polyphosphoric acids
• CPC Class Codes: C07H 21/02 with ribosyl as saccharide radical

Abstract

The present invention provides a method for preparing nucleotide oligomers, including (a) coupling a nucleotide dimer or nucleotide trimer to a nucleoside attached to solid supports or to universal solid supports as a starting material; (b) sequentially coupling nucleotide monomers to the resulting structures of Step (a) to prepare a nucleotide oligomer; and (c) removing the nucleotide oligomers from the solid supports.

The method of the present invention provides nucleotide oligomers having 15-20% higher purity than the conventional art. The present invention enables the efficient and inexpensive synthesis of nucleotide oligomers with high purity within a shorter period of time.

Claims

1. A method for preparing nucleotide oligomers, comprising:

(a) coupling a nucleotide dimer or nucleotide trimer to a nucleoside attached to solid supports or to universal solid supports as a starting material;

(b) sequentially coupling nucleotide monomers to the resulting structures of Step (a) to prepare nucleotide oligomers; and

(c) removing the nucleotide oligomers from the solid supports.

2. The method of claim 1, wherein the method includes:

(a) coupling a nucleotide dimer to a nucleoside attached to solid supports or to a universal solid supports as a starting material;

(b) sequentially coupling nucleotide monomers to the resulting structures of Step (a) to prepare nucleotide oligomers; and

(c) removing the nucleotide oligomers from the solid supports.

3. The method of claim 1, wherein the method includes:

(a) coupling a nucleotide trimer to a nucleoside attached to solid supports or to a universal solid supports as a starting material;

(b) sequentially coupling nucleotide monomers to the resulting structures of Step (a) to prepare nucleotide oligomers; and

(c) removing the nucleotide oligomers from the solid supports.

4. The method of claim 1, wherein the nucleotide is a ribonucleotide.

5. The method of claim 1, wherein the nucleotide monomer is a ribonucleoside phosphoramidite.

6. The method of claim 1, wherein the nucleotide oligomer is one containing at least one ribonucleotide selected from 2′

-O′

-halogen ribonucleotide, 2′

-amino ribonucleotide, 2′

-O-alkyl ribonucleotide and 2′

-O-alkoxy ribonucleotide.

7. The method of claim 1, wherein the nucleotide oligomer has a phosphodiester, phosphoramidate, alkylphosphoramidate, alkylphosphonate, phosphorothioate, alkylphosphotriester, or alkylphosphonothioate linkage.

8. The method of claim 7, wherein the nucleotide oligomer has a phosphodiester or phosphoramidate linkage.

9. The method of claim 1, wherein the nucleotide dimer is prepared by a reaction including coupling of a compound of Formula 1 with a compound of Formula 2:

10. The method of claim 1, wherein the nucleotide dimer is prepared by a reaction including coupling of a compound of Formula 3 with a compound of Formula 2:

11. The method of claim 1, wherein the nucleotide trimer is prepared by a reaction including:

(a) reacting a ribonucleotide dimer of Formula 4 with an acid to remove R₁ of Formula 4; and

(b) coupling the resulting product of Step (a) with a ribonucleoside 3′

-phosphoramidite to prepare a ribonucleotide trimer

12. The method of claim 11, wherein the acid of Step (a) is benzenesulfonic acid.

15. The method of claim 4, wherein the ribonucleotide includes a sugar in which a methyl or an ethyl is bonded to —

OH of a C2 carbon.

16. The method of claim 4, wherein the ribonucleotide includes a sugar in which a flora (—

F) or an amino is boned to a C2 carbon instead of —

OH.

17. The method of claim 6, wherein the 2′

-O-halogen ribonucleotide includes 2′

-O-fluoro-ribonucleotide.

18. The method of claim 6, wherein the 2′

-O-alkyl ribonucleotide includes 2′

-O-methyl ribonucleotide.

19. The method of claim 6, wherein the 2′

-O-alkoxy ribonucleotide includes 2′

-O-methoxy-ribonucleotide.

13. A ribonucleotide dimer represented by Formula 5:

14. A ribonucleotide trimer represented by Formula 6: