Derivatized oligonucleotides having improved uptake and other properties

• US 7,122,649 B2
• Filed: 01/09/2004
• Issued: 10/17/2006
• Est. Priority Date: 10/23/1992
• Status: Expired due to Fees

• Current Assignee: ISIS Pharmaceuticals Incorporated
• Original Assignee: ISIS Pharmaceuticals Incorporated
• Inventors: Manoharan, Muthiah, Cook, Phillip Dan, Bennett, Clarence Frank
• Primary Examiner(s): Riley, Jezia
• Application Number:
  US10/755,109
• Publication Number:
  US 20050158727A1
• Time in Patent Office: 1,012 Days
• Field of Search: 536/23.1, 536/25.3, 536/26.6, 435/6, 514/1, 514/44
• US Class Current: 536/23.1
• CPC Class Codes: C07H 21/04 with deoxyribosyl as saccharide radical
• CPC Class Codes: C07J 1/00 Normal steroids containing carbon, hydrogen, halogen or oxygen, not substituted in position 17 beta by a carbon atom, e.g. estrane, androstane
• CPC Class Codes: C12N 2710/20022 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: H04J 2203/0042 Ring
• CPC Class Codes: H04J 2203/006 Fault tolerance and recovery

Abstract

Linked nucleosides having at least one functionalized nucleoside that bears a substituent such as a steroid molecule, a reporter molecule, a non-aromatic lipophilic molecule, a reporter enzyme, a peptide, a protein, a water soluble vitamin, a lipid soluble vitamin, an RNA cleaving complex, a metal chelator, a porphyrin, an alkylator, a pyrene, a hybrid photonuclease/intercalator, or an aryl azide photo-crosslinking agent exhibit increased cellular uptake and other properties. The substituent can be attached at the 2′-position of the functionalized nucleoside via a linking group. If at least a portion of the remaining liked nucleosides are 2′-deoxy-2′-fluoro, 2′-O-methoxy, 2′-O-ethoxy, 2′-O-propoxy, 2′-O-aminoalkoxy or 2′-O-allyloxy nucleosides, the substituent can be attached via a linking group at any of the 3′ or the 5′ positions of the nucleoside or on the heterocyclic base of the nucleoside or on the inter-nucleotide linkage linking the nucleoside to an adjacent nucleoside.

Claims

1. A compound comprising a plurality of linked nucleosides including a plurality of 2′

-deoxy′

-2′

-fluoro, 2′

-O—

C₁–

C₂₀-alkyl, 2′

-O—

C₂–

C₂₀-alkenyl, 2′

-O—

C₂–

C₂₀-alkynyl, 2′

-S—

C₁–

C₂₀-alkyl, 2′

-S—

C₂–

C₂₀-alkenyl, 2′

-S—

C₂–

C₂₀-alkynyl, 2′

-NH—

C₁–

C₂₀-alkyl, 2′

-NH—

C₂–

C₂₀-alkenyl or 2′

-NH—

C₂–

C₂₀-alkynyl nucleosides; and

a 5′

-O-terminal nucleoside having a conjugate group linked via a linker to the 5′

-O-position of the nucleoside,

provided that the compound does not include sulfamate or sulfamide internucleotide linkages.

2. The compound of claim 1 including a plurality of 2′

-methoxy nucleosides.

3. The compound of claim 2 including a bifunctional linker linking said conjugate group to said 5′

-O-position of said 5′

-O-terminal nucleoside.

4. A compound comprising a plurality of linked nucleosides including a plurality of 2′

-deoxy′

-2′

-fluoro, 2′

-O—

C₁–

C₂₀-alkyl, 2′

-O—

C₂–

C₂₀-alkenyl, 2′

-O—

C₂–

C₂₀-alkynyl, 2′

-S—

C₁–

C₂₀-alkyl, 2′

-S—

C₂–

C₂₀-alkenyl, 2′

-S—

C₂–

C₂₀-alkynyl, 2′

-NH—

C₁–

C₂₀-alkyl, 2′

-NH—

C₂–

C₂₀-alkenyl or 2′

-NH—

C₂–

C₂₀-alkynyl nucleosides; and

a 5′

-O-terminal nucleoside having a conjugate group linked via a bifunctional linking group to the 5′

-O-position of the nucleoside,

provided that the compound does not include sulfamate or sulfamide internucleotide linkages.

5. The compound of claim 4 including a plurality of 2′

-methoxy nucleosides.

6. A compound comprising a plurality of linked nucleosides including a plurality of 2′

-O—

C₁–

C₂₀-alkyl nucleosides; and

a 5′

-O-terminal nucleoside having a conjugate group linked via a bifunctional linking group to the 5′

-O-position of the nucleoside,

provided that the compound does not include sulfamate or sulfamide internucleotide linkages.

7. The compound of claim 6 wherein said 2′

-O—

C₁–

C₂₀-alkyl nucleosides are 2′

-methoxy nucleosides.