Derivatized oligonucleotides having improved uptake and other properties

• US 6,831,166 B2
• Filed: 02/11/2002
• Issued: 12/14/2004
• 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:
  US10/073,718
• Publication Number:
  US 20020177150A1
• Time in Patent Office: 1,037 Days
• Field of Search: 435/5, 435/6, 536/23.1, 536/24.3, 536/25.3, 536/26.6
• 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, wherein at least one of said nucleosides is functionalized at its 2′

-position by attachment of a steroid molecule, a reporter molecule, a water soluble vitamin, an RNA cleaving complex, a porphyrin, an alkylator, a hybrid photonuclease, or an aryl azide photo-crosslinking agent.

2. A method of effecting cellular uptake of a compound having a plurality of linked nucleosides, comprising contacting an organism with said compound wherein said compound includes at least one heterocyclic base functionalized nucleoside having a steroid molecule, an enzyme, a peptide, a protein, or a vitamin, linked to the heterocyclic base of the nucleoside.

3. The method of claim 2 wherein said heterocyclic base functionalized nucleoside is functionalized with a steroid molecule.

4. The method of claim 2 wherein said heterocyclic base functionalized nucleoside is functionalized with an enzyme, a peptide or a protein.

5. The method of claim 2 wherein said heterocyclic base functionalized nucleoside is functionalized with a vitamin.

6. A method of effecting cellular uptake of a compound having a plurality of linked nucleosides, comprising contacting an organism with said compound wherein said compound includes at least one inter-strand nucleoside having a steroid molecule, an enzyme, a peptide, a protein, or a vitamin linked to an inter-strand linkage linking said inter-strand nucleoside to an adjacent nucleoside.

7. The method of claim 6 wherein said inter-strand nucleoside has a steroid molecule linked to an inter-strand linkage linking said inter-strand nucleoside to an adjacent nucleoside.

8. The method claim 7 wherein said inter-strand linkage is not the inter-strand linkage between the first and the second nucleosides at the 3′

terminus or the 5′

terminus of said oligonucleotide.

9. The method of claim 6 wherein said inter-strand nucleoside has an enzyme, a peptide or a protein linked to an inter-strand linkage linking said inter-strand nucleoside to an adjacent nucleoside.

10. The method claim 6 wherein said inter-strand nucleoside has a vitamin linked to an inter-strand linkage linking said inter-strand nucleoside to an adjacent nucleoside.