Chimeric immunomodulatory compounds and methods of using the same—I

• US 7,255,868 B2
• Filed: 06/21/2002
• Issued: 08/14/2007
• Est. Priority Date: 06/21/2001
• Status: Active Grant

• Current Assignee: Dynamax Technologies Limited
• Original Assignee: Dynamax Technologies Limited
• Inventors: Fearon, Karen L., Dina, Dino, Tuck, Stephen F.
• Primary Examiner(s): Duffy; Patricia A.
• Assistant Examiner(s): GANGLE, BRIAN J
• Application Number:
  US10/176,883
• Publication Number:
  US 20030175731A1
• Time in Patent Office: 1,880 Days
• Field of Search: 536/22.1, 536/23.1
• US Class Current: 424/280.1
• CPC Class Codes: A61K 2039/55561 CpG containing adjuvants; Oligonucleotide containing adjuvants
• CPC Class Codes: A61K 39/12 Viral antigens
• CPC Class Codes: A61K 39/292 Serum hepatitis virus, hepatitis B virus, e.g. Australia antigen
• CPC Class Codes: A61K 47/54 the modifying agent being an organic compound
• CPC Class Codes: A61K 47/549 Sugars, nucleosides, nucleotides or nucleic acids
• CPC Class Codes: A61P 11/06 Antiasthmatics
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 31/10 Antimycotics
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 33/00 Antiparasitic agents
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: A61P 37/08 Antiallergic agents antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14
• CPC Class Codes: C07H 15/18 Acyclic radicals, substituted by carbocyclic rings
• 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: C12N 15/117 Nucleic acids having immunomodulatory properties, e.g. containing CpG-motifs
• CPC Class Codes: C12N 2310/17 Immunomodulatory nucleic acids
• CPC Class Codes: C12N 2310/315 : Phosphorothioates
• CPC Class Codes: C12N 2310/332 : Abasic residue
• CPC Class Codes: C12N 2310/351 : Conjugate
• CPC Class Codes: C12N 2310/3519 : Fusion with another nucleic acid
• CPC Class Codes: C12N 2310/52 : branched
• CPC Class Codes: C12N 2730/10134 : Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein

Abstract

The invention provides immunomodulatory compounds and methods for immunomodulation of individuals using the immunomodulatory compounds.

Claims

1. A chimeric immunostimulatory compound (CIC) comprising the structure N₁-S₁-N₂-N₃,

wherein N₁, N₂, and N₃ are independently selected nucleic acid moieties comprising 3 to 7 bases, at least one of N₁, N₂, and N₃ has the sequence 5′

-[(X)_0-2]TCG[(X)_2-4]-3′

, wherein each X is an independently selected nucleotide;

wherein S₁ is a non-nucleic acid spacer moiety covalently bound to N₁ and N₂;

S₂ is a non-nucleic acid spacer moiety covalently bound to N₂ and N₃;

S₁ and S₂ are the same or different;

each of S₁ and S₂ comprises hexaethylene glycol (HEG), triethylene glycol (TEG), propyl, or butyl; and

wherein said CIC has at least one immunostimulatory activity selected from the group consisting of (i) the ability to stimulate interferon-gamma (IFN-γ

) production by human peripheral blood mononuclear cells and (ii) the ability to stimulate interferon-alpha (IFN-α

) production by human peripheral blood mononuclear cells.

2. The CIC of claim 1 wherein at least one of S₁ and S₂ is a HEG spacer moiety.

3. The CIC of claim 2 wherein both S₁ and S₂ are HEG spacer moieties.

4. The CIC of claim 1 wherein at least one of N₁ and N₃ has a sequence 5′

-TCG[(X)_2-4]-3′

.

5. The CIC of claim 1 wherein at least one of N₁ and N₃ has a sequence 5′

-TCG(A/T)[(X)_1-3]-3′

.

6. The CIC of claim 5 wherein at least one of N₁ and N₃ has a sequence 5′

-TCG(A/T)CG(A/T)-3′

.

7. The CIC of claim 6 wherein at least one nucleic acid moiety comprises the sequence 5′

-TCGACGT-3′

or 5′

-TCGTCGA-3′

.

8. The CIC of claim 3 wherein at least one of N₁ and N₃ has a sequence 5′

-TCG(A/T)[(X)_1-3]-3′

.

9. The CIC of claim 8 wherein at least one of N₁ and N₃ has a sequence 5′

-TCG(A/T)CG(A/T)-3′

.

10. The CIC of claim 9 wherein at least one nucleic acid moiety comprises the sequence 5′

-TCGACGT-3′

or 5′

-TCGTCGA-3′

.

11. The CIC of claim 1, where the linkages between nucleotides of the nucleic acid moieties and between the nucleic acid moieties and the nonnucleic acid spacer moieties comprise one or more linkages selected from the group consisting of phosphodiester linkages, phosphorothioate ester linkages, and phosphorodithioate ester linkages.

12. A composition comprising a CIC of claim 1 and a pharmaceutically acceptable excipient.

13. A composition comprising a CIC of claim 1 and further comprising an antigen.

14. A composition comprising a CIC of claim 1 and further comprising a cationic microsphere.

15. The composition of claim 14 wherein the microsphere comprises a polymer of lactic acid and glycolic acid.

16. A CIC of claim 1 comprising a reactive linking group.

17. A CIC of claim 16 comprising a reactive thio group.

18. A CIC of claim 1 linked to a polypeptide.

19. The CIC of claim 11 having a structure selected from the group consisting of

5′

-TCGTCGA-3′

-HEG-5′

-ACGTTCG-3′

-HEG-5′

-AGATGAT-3′

5′

-TCGTCG-3′

-HEG-5′

-ACGTTCG-3′

-HEG-5′

-AGATGAT-3′

5′

-TCGTCGA-3′

-HEG-5′

-TCGTCGA-3′

-HEG-5′

-TCGTCGA-3′

5′

-TCGTCG-3′

-HEG-5′

-TCGTCG-3′

-HEG-5′

-TCGTCG-3′

5′

-TCGTCG-3′

-HEG-5′

-AACGTT-3′

-HEG-5′

-AGATGAT-3′

5′

-TCGTCG-3′

-HEG-5′

-ACGTTCG-3′

-HEG-5′

-AGATGAT-3′

-TEG

HEG-5′

-TCGTCG-3′

-HEG-5′

-ACGTTCG-3′

-HEG-5′

-AGATGAT-3′

-TEG

5′

-TCGTTTT-3′

-HEG-5′

-TCGTTTT-3′

-HEG-5′

-TCGTTTT-3′

5′

-TCGTCGT-3′

-HEG-5′

-TCGTCGT-3′

-HEG-5′

-TCGTCGT-3′

5′

-TCGAGAT-3′

-HEG-5′

-TCGAGAT-3′

-HEG-5′

-TCGAGAT-3′

5′

-TCGTCGT-3′

-HEG-5′

-TGTCGTT-3′

-HEG-5′

-TGTCGTT-3′

5′

-TCGTCGA-3′

-HEG-5′

-ACGTTCG-3′

-HEG-5′

-TCGTCGA-3′

5′

-TCGTCGA-3′

-HEG-5′

-ACGTTCG-3′

-HEG-5′

-GGGGGG-3′

5′

-TCGAACG-3′

-HEG-5′

-TCGAACG-3′

-HEG-5′

-TCGAACG-3′

5′

-TCGACGT-3′

-HEG-5′

-TCGACGT-3′

-HEG-5′

-TCGACGT-3′

5′

-TCGTCGA-3′

-HEG-5′

-AACGTTC-3′

-HEG-5′

-AGATGAT-3′

5′

-TCGTCGA-3′

-HEG-5′

-AACGTTC-3′

-HEG-5′

-TCGTCGA-3′

5′

-TCGTCGA-3′

-HEG-5′

-AGATGAT-3′

-HEG-5′

-ACGTTCG-3′

5′

-TCGACTC-3′

-HEG-5′

-TCGAGCG-3′

-HEG-5′

-TTCTCTT-3′

5′

-TCGTCGA-3′

-HEG-5′

-TCGTCGA-3′

-HEG-3′

-AGCTGCT-5′

5′

-TCGAT-3′

-HEG-5′

-TCGAT-3′

-HEG-5′

-TCGAT-3′

-HEG-5′

-TCGAT-3′

5′

-TCGTCGA-3′

-HEG-5′

-TCGTCGA-3′

-HEG-5′

-AACGTTC-3′

-HEG-5′

-AGAT-3′

5′

-TCGACGT-3′

-HEG-5′

-TCGACGT-3′

-HEG-5′

-TCGACGT-3′

-HEG-5′

-TCGACGT-3′

5′

-TCGATTT-3′

-HEG-5′

-TCGATTT-3′

-HEG-5′

-TCGATTT-3′

5′

-TCGCTTT-3′

-HEG-5′

-TCGCTTT-3′

-HEG-5′

-TCGCTTT-3′

5′

-TCGGTTT-3′

-HEG-5′

-TCGGTTT-3′

-HEG-5′

-TCGGTTT-3′

.

20. A composition comprising a CIC of claim 19 and a pharmaceutically acceptable excipient.

21. A composition comprising a CIC of claim 19 and further comprising an antigen.

22. A composition comprising a CIC of claim 19 and further comprising a cationic microsphere.

23. The composition of claim 22 wherein the microsphere comprises a polymer of lactic acid and glycolic acid.

24. A CIC of claim 19 further comprising a reactive thio group.

25. A CIC of claim 19 linked to a polypeptide.