Methods of synthesis and use

• US 6,235,886 B1
• Filed: 03/09/1995
• Issued: 05/22/2001
• Est. Priority Date: 09/03/1993
• Status: Expired due to Fees

• Current Assignee: ISIS Pharmaceuticals Incorporated (Ionis Pharmaceuticals, Inc.)
• Original Assignee: ISIS Pharmaceuticals Incorporated (Ionis Pharmaceuticals, Inc.)
• Inventors: Manoharan, Muthiah, Cook, P. Dan
• Primary Examiner(s): FREDMAN, JEFFREY NORMAN
• Application Number:
  US08/397,277
• Time in Patent Office: 2,266 Days
• Field of Search: 536/23.1, 536/25.3, 436/6
• US Class Current: 536/22.1
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids

Abstract

Oligonucleotide and nucleotide amine analogs and methods of preparing and using these compounds are provided by the present invention.

Claims

1. A compound having the structure:

wherein;

B is a purine or pyrimidine heterocyclic base;

R₈ and R₉ are, independently, H, PO₂⁻

, a nucleotide, an oligonucleotide or an amine-containing species, and at least one of R₈ and R₉ is a purine containing oligonucleotide; and

at least one of R₉ and A is a species comprising the formula L₁-L₂-polyamine wherein L₁ is an amino linker comprising an amino group, and L₂ is a heterobifunctional linker;

provided that;

if R₈ is not a purine containing oligonucleotide then;

R₈ is a nucleotide or PO₂⁻

;

or

R₉ is H or a nucleotide; and

if A is not L₁-L₂-polyamine, then A is H or OH.

2. The compound of claim 1 wherein the amine-containing species comprises at least one nitrogen atom having a free electron pair.

3. The compound of claim 1 further including one or more of reporter groups, alkylating agents, intercalating agents, cell receptor binding molecules, steroids, peptides, crown amines, porphyrins, or cross-linking agents attached to at least one of the nitrogen atoms of said amine-containing species.

4. The compound of claim 1 in a pharmaceutically acceptable carrier.

5. The compound of claim 1 wherein R₈ is an amine-containing species.

6. The compound of claim 1 wherein R₉ is an amine-containing species.

7. The compound of claim 1 wherein R₈ is L₁-L₂-amine.

8. The compound of claim 1 wherein R₉ is L₁-L₂-amine.

9. The compound of claim 1 wherein A is L₁-L₂-amine.

10. The compound of claim 1 wherein said heterobifunctional linker is bis-(maleimido)-methyl ether, disuccinimidyl suberate , 3-maleimidobenzoyl-N-hydroxy-succinimide, maleimidohexanoyl-N-hydroxyl-succinimide, and N-succinimidyl-3-(2-pyridyldithio)propionate.

11. The compound of claim 1 wherein said polyamine is selected from primary amines, hydrazines, semicarbazines, and thiosemicarbazines.

12. The compound of claim 1 wherein at least one nitrogen atom of said amine has a free electron pair.

13. The compound of claim 1 wherein wherein said amine contains non-amide nitrogen atoms.

14. The compound of claim 1 wherein said amine has the formula H₂N[(CH₂)_nNH]_m—

wherein n is 2-8 and m is 1-10.

15. The compound of claim 1 wherein said amine is cyclic.