Peptide nucleic acids having 2,6-diaminopurine nucleobases

• US 6,613,873 B1
• Filed: 06/21/1999
• Issued: 09/02/2003
• Est. Priority Date: 05/24/1991
• Status: Expired due to Fees

• Current Assignee: ISIS Pharmaceuticals Incorporated (Ionis Pharmaceuticals, Inc.)
• Original Assignee: ISIS Pharmaceuticals Incorporated (Ionis Pharmaceuticals, Inc.)
• Inventors: Egholm, Michael, Buchardt, Ole, Berg, Rolf Henrik, Nielsen, Peter Eigil
• Primary Examiner(s): Marschel, Ardin H.
• Application Number:
  US09/337,304
• Time in Patent Office: 1,534 Days
• Field of Search: 435/6, 514/2, 514/44, 530/300, 530/350
• US Class Current: 530/300
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• 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
• CPC Class Codes: C07K 14/003 Peptide-nucleic acids (PNAs)
• CPC Class Codes: C07K 5/06026 the side chain containing 0 or 1 carbon atom, i.e. Gly or Ala
• CPC Class Codes: C07K 5/06139 with the first amino acid being heterocyclic
• CPC Class Codes: C07K 7/06 having 5 to 11 amino acids
• CPC Class Codes: C07K 7/08 having 12 to 20 amino acids gastrins C07K14/595; somatostatins C07K14/655; melanotropins C07K14/68
• CPC Class Codes: C12Q 1/68 involving nucleic acids
• CPC Class Codes: C12Q 1/6813 Hybridisation assays
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: C12Q 2535/107 Maxam and Gilbert method, i.e. sequential release and detection of nucleotides

Abstract

A novel class of compounds, known as peptide nucleic acids, bind complementary DNA and RNA strands more strongly than a corresponding DNA strand, and exhibit increased sequence specificity and binding affinity. The peptide nucleic acids of the invention comprise ligands selected from a group consisting of naturally-occurring nucleobases and non-naturally-occurring nucleobases attached to a polyamide backbone. Some PNAs of the invention also contain C₁-C₈ alkylamine side chains.

Claims

1. A peptide nucleic acid having formula:

wherein;

each L is independently selected from a group consisting of naturally-occurring nucleobases and non-naturally-occurring nucleobases, at least one of said L being a 2,6-diaminopurine nucleobase;

each R^7′

is independently hydrogen or C₁-C₈ alkylamine;

R^h is OH, NH₂ or NHLysNH₂;

Rⁱ is H, COCH₃ or t-butoxycarbonyl; and

n is an integer from 1 to 30.

2. The peptide nucleic acid of claim 1 wherein said R^7′

is C₃-C₆ alkylamine.

3. The peptide nucleic acid of claim 2 wherein said R^7′

is C₄-C₅ alkylamine.

4. The peptide nucleic acid of claim 3 wherein said R^7′

is butylamine.

5. The peptide nucleic acid of claim 1 wherein at least one of said R^7′

is C₁-C₈ alkylamine.

6. The peptide nucleic acid of claim 5 wherein at least one of said R^7′

is C₃-C₆ alkylamine.

7. The peptide nucleic acid of claim 6 wherein at least one of said R^7′

is C₄-C₅ alkylamine.

8. The peptide nucleic acid of claim 7 wherein at least one of said R^7′

is butylamine.

9. The peptide nucleic acid of claim 7 wherein substantially all of said R^7′

is butylamine.

10. The peptide nucleic acid of claim 1 wherein the carbon atom to which the substituent R^7′

is attached is stereochemically enriched.

11. The peptide nucleic acid of claim 10 wherein said stereochemical enrichment is of the R configuration.

12. The peptide nucleic acid for claim 1 wherein said peptide nucleic acid is derived from an amino acid.

13. The peptide nucleic acid for claim 12 wherein said peptide nucleic acid is derived from D-lysine.

23. A pharmaceutical composition comprising a peptide nucleic acid according to claim 1 and at least one pharmaceutically effective carrier, binder, thickener, diluent, buffer, preservative, or surface active agent.

14. A compound having formula:

wherein;

L is a 2,6-diaminopurine nucleobase;

R^7′

is hydrogen or C₁-C₈ alkylamine;

E is COOH or an activated or protected derivative thereof; and

Z is NH₂ or NHPg, where Pg is an amino-protecting group.

15. The compound of claim 14 wherein said R^7′

is C₃-C₆ alkylamine.

16. The compound of claim 15 wherein said R^7′

is C₄-C₅ alkylamine.

17. The compound of claim 16 wherein said R^7′

is butylamine.

18. The compound of claim 14 wherein the carbon atom to which the substituent R^7′

is attached is stereochemically enriched.

19. The compound of claim 18 wherein said stereochemical enrichment is of the R configuration.

20. The compound of claim 14 wherein said compound is derived from an amino acid.

21. The compound of claim 20 wherein said compound is derived from D-lysine.

22. The compound of claim 14 wherein said Pg is t-butoxycarbonyl.