2'-O-modified RNA

• US 8,822,671 B2
• Filed: 11/28/2011
• Issued: 09/02/2014
• Est. Priority Date: 11/30/2010
• Status: Active Grant

• Current Assignee: Chiralgen, Ltd., University of Tokyo
• Original Assignee: Chiralgen, Ltd., University of Tokyo
• Inventors: Shimizu, Mamoru, Wada, Takeshi, Arai, Kouichiro
• Primary Examiner(s): LEWIS, PATRICK T
• Application Number:
  US13/990,441
• Publication Number:
  US 20130253178A1
• Time in Patent Office: 1,009 Days
• Field of Search: None
• US Class Current: 536/25.3
• CPC Class Codes: C07H 1/00 Processes for the preparation of sugar derivatives
• CPC Class Codes: C07H 19/06 Pyrimidine radicals
• 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 21/02 with ribosyl as saccharide radical
• CPC Class Codes: C07H 23/00 Compounds containing boron, silicon, or a metal, e.g. chelates, vitamin B12 esters with inorganic acids C07H11/00; metal salts, see parent compounds
• CPC Class Codes: Y02P 20/55 Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Abstract

A 2′-modified ribonucleoside having an alkoxymethyl protective group can be imparted with a high duplex-forming ability by introducing, as a substituent, a halogen atom into the protective group moiety. A modified form of RNA having a halogen-substituted alkoxymethyl protective group exhibits a high duplex-forming ability that is comparable to the duplex-forming ability of a 2′-O-methyl modified nucleic acid.

Claims

1. A ribonucleoside, a ribonucleotide, or a derivative thereof having a protective group at the 2′

-position, wherein the protective group is represented by the following Formula (I);

2. The ribonucleoside, the ribonucleotide, or the derivative thereof having a protective group at the 2′

-position according to claim 1,

wherein R³ and R⁴ may be identical or different, and each represents a hydrogen atom, a methyl group or an ethyl group,

R⁵ and R⁶ may be identical or different, and each represents a hydrogen atom, a halogen atom, or a C_1-3 haloalkyl group, and

R⁷ represents a halogen atom.

3. The ribonucleoside, the ribonucleotide, or the derivative thereof having a protective group at the 2′

-position according to claim 1,

wherein all of R¹ to R⁴ represent a hydrogen atom,

R⁵ and R⁶ may be identical or different, and each represents a hydrogen atom, a fluorine atom, a chlorine atom or a bromine atom, and

R⁷ represents a fluorine atom, a chlorine atom or a bromine atom.

4. The ribonucleoside, the ribonucleotide, or the derivative thereof having a protective group at the 2′

-position according to claim 1,

wherein all of R¹ to R⁵ represent a hydrogen atom,

R⁶ represents a hydrogen atom, a fluorine atom or a chlorine atom, and

R⁷ represents a fluorine atom or a chlorine atom.

5. The ribonucleoside, the ribonucleotide, or the derivative thereof having a protective group at the 2′

-position according to claim 1,

wherein all of R¹ to R⁵ represent a hydrogen atom,

R⁶ represents a hydrogen atom, or a chlorine atom, and

R⁷ represents a chlorine atom.

6. The ribonucleoside, the ribonucleotide, or the derivative thereof having a protective group at the 2′

-position according to claim 1, which is

2′

-O-(2-chloro)ethoxymethyl-3′

,5′

-O-(1,1,3,3-tetraisopropyldisiloxane-1,3diyl) nucleic acid base B,

2′

-O-(2,2-dichloro)ethoxymethyl-3′

,5′

-O-(1,1,3,3-tetraisopropyldisiloxane-1,3diyl) nucleic acid base B,

2′

-O-(2,2,2-trichloro)ethoxymethyl-3′

,5′

-O-(1,1,3,3-tetraisopropyldisiloxane-1,3diyl) nucleic acid base B,

2′

-O-(2,2-difluoro)ethoxymethyl-3′

,5′

-O-(1,1,3,3-tetraisopropyldisiloxane-1,3diyl) nucleic acid base B,

2′

-O-(2,2,2-trifluoro)ethoxymethyl-3′

,5′

-O-(1,1,3,3-tetraisopropyldisiloxane-1,3diyl) nucleic acid base B,

2′

-O-(2-chloro)ethoxymethyl-5′

-O-(4,4′

-dimethoxytrityl) nucleic acid base B,

2′

-O-(2,2-dichloro)ethoxymethyl-5′

-O-(4,4′

-dimethoxytrityl) nucleic acid base B,

2′

-O-2,2,2-trichloroethoxymethyl-5′

-O-(4,4′

-dimethoxytrityl) nucleic acid base B,

2′

-O-2,2-difluoroethoxymethyl-5′

-O-(4,4′

-dimethoxytrityl) nucleic acid base B,

2′

-O-2,2,2-trifluoroethoxymethyl-5′

-O-(4,4′

-dimethoxytrityl) nucleic acid base B,

2′

-O-(2-chloro)ethoxymethyl-5′

-O-(4,4′

-dimethoxytrityl) nucleic acid base B 3′

-(2-cyanoethyldiisopropyl phosphoramidite),

2′

-O-(2,2-dichloro)ethoxymethyl-5′

-O-(4,4′

-dimethoxytrityl) nucleic acid base B 3′

-(2-cyanoethyldiisopropyl phosphoramidite),

2′

-O-(2,2,2-trichloro)ethoxymethyl-5′

-O-(4,4′

-dimethoxytrityl) nucleic acid base B 3′

-(2-cyanoethyldiisopropyl phosphoramidite),

2′

-O-(2,2-difluoro)ethoxymethyl-5′

-O-(4,4′

-dimethoxytrityl) nucleic acid base B 3′

-(2-cyanoethyldiisopropyl phosphoramidite),

2′

-O-(2,2,2-trifluoro)ethoxymethyl-5′

-O-(4,4′

-dimethoxytrityl) nucleic acid base B 3′

-(2-cyanoethyldiisopropyl phosphoramidite),

2′

-O-(2-bromo)ethoxymethyl-3′

,5′

-O-(1,1,3,3-tetraisopropyldisiloxane-1,3diyl) nucleic acid base B,

2-O-(2,2-dibromo)ethoxymethyl-3′

,5′

-O-(1,1,3,3-tetraisopropyldi siloxane-1,3 diyl) nucleic acid base B,

2′

-O-(2,2,2-tribromo)ethoxymethyl-3′

,5′

-O-(1,1,3,3-tetraisopropyldisiloxane-1,3 diyl) nucleic acid base B,

2′

-O-(2-bromo)ethoxymethyl-5′

-O-(4,4′

-dimethoxytrityl) nucleic acid base B 3′

-(2-cyanoethyldiisopropyl phosphoramidite),

2′

-O-(2,2-dibromo)ethoxymethyl-5′

-O-(4,4′

-dimethoxytrityl) nucleic acid base B 3′

-(2-cyanoethyldiisopropyl phosphoramidite), or

2′

-O-(2,2,2-tribromo)ethoxymethyl-5′

-O-(4,4′

-dimethoxytrityl) nucleic acid base B 3′

-(2-cyanoethyldiisopropyl phosphoramidite),

wherein the nucleic acid base B is adenosine, guanosine, cytidine or uridine.

7. The ribonucleoside, the ribonucleotide, or the derivative thereof having a protective group at the 2′

-position according to claim 1, which is

2′

-O-(2-chloro)ethoxymethyl-5′

-O-(4,4′

-dimethoxytrityl) nucleic acid base B 3′

-(2-cyanoethyldiisopropyl phosphoramidite), or

2′

-O-(2,2-dichloro)ethoxymethyl-5′

-O-(4,4′

-dimethoxytrityl) nucleic acid base B 3′

-(2-cyanoethyldiisopropyl phosphoramidite);

wherein the nucleic acid base B is adenosine, guanosine, cytidine or uridine.

8. A method of manufacturing the ribonucleoside, the ribonucleotide, or the derivative thereof represented by the Formula (I) according to claim 1, the method comprising:

reacting a compound represented by the following Formula (III) with a compound represented by the following Formula (II);

9. The method according to claim 8, wherein the reacting the compound represented by Formula (III) with the compound represented by Formula (II) produces a compound represented following Formula (IV), the method further comprising:

reacting the compound represented by Formula (IV) with a C_1-3 alcohol to obtain a compound represented by the following Formula (V); and

reacting the compound represented by Formula (V) with dimethoxytriphenylmethyl halide, to obtain a compound represented by Formula (VI).

10. The method according to claim 9, further comprising, after obtaining the compound represented by Formula (VI):

reacting the compound represented by Formula (VI) with a compound represented by the following Formula (VII) to obtain a compound represented by the following Formula (VIII);