Chiral nucleic acid adjuvant

• US 9,617,547 B2
• Filed: 07/12/2013
• Issued: 04/11/2017
• Est. Priority Date: 07/13/2012
• Status: Active Grant

• Current Assignee: Wave Life Sciences Japan, Inc.
• Original Assignee: Shin Nippon Biomedical Laboratories Limited, Wave Life Sciences Japan KK (WAVE Life Sciences Ltd.)
• Inventors: Gemba, Takefumi
• Primary Examiner(s): Minnifield, Nita M
• Application Number:
  US14/414,551
• Publication Number:
  US 20150166999A1
• Time in Patent Office: 1,369 Days
• Field of Search: None
• CPC Class Codes: A61K 2039/55561 CpG containing adjuvants; Oligonucleotide containing adjuvants
• CPC Class Codes: A61K 39/39 characterised by the immunostimulating additives, e.g. chemical adjuvants
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• 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 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• 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

Abstract

Problem

The purpose of the present invention is to provide: a stereo isomer of a novel CpG oligonucleotide, which has excellent stability; and a CpG oligonucleotide which has a capability of producing interferon-α (IFNα).

Solution

The present invention relates to an oligonucleotide which contains two to four sequences each represented by the formula 5′-X₁X₂CpGX₃X₄-3′ (formula (I)) and has a length of 14 to 32 nucleotides. In formula (I), CpG represents a non-methylated CpG residue having a phosphate skeleton modification, X₁X₂ represents any one of AA, AT, GA and GT, and X₃X₄ represents any one of TT, AT, AC and CG. The oligonucleotide has at least one phosphate skeleton modification at an S-form stereoisomer located at a site other than the CpG.

Claims

1. An oligonucleotide which comprises two to four sequences each represented by 5′

-X₁X₂CpGX₃X₄-3′ and

has a length of 14 to 32 nucleotides,

wherein the CpG is non-methylated CpG without a modified phosphate backbone,

wherein X₁X₂ is AA, AT, GA, or GT, and contains no modified phosphate backbones,

wherein X₃X₄ is TT, AT, AC, TC, or CG, and contains no modified phosphate backbones, and

the oligonucleotide comprises at least one modified phosphate backbone at a site other than the parts represented by 5′

-X₁X₂CpGX₃X₄-3′

.

2. An oligonucleotide which comprises two to four sequences each represented by 5′

-X₁X₂CpGX₃X₄-3′ and

has a length of 14 to 32 nucleotides,

wherein the CpG is non-methylated CpG without a modified phosphate backbone,

wherein X₁X₂ is AA, AT, GA, or GT, and may comprise a modified phosphate backbone,

wherein X₃X₄ is TT, AT, AC, TA, TC, or CG, and may comprise a modified phosphate backbone, and

wherein the oligonucleotide comprises at least one Sp modified phosphate backbone at a site other than the parts represented by 5′

-X₁X₂CpGX₃X₄-3′

.

3. An oligonucleotide which comprises two to four sequences each represented by 5′

-X₁X₂CpGX₃X₄-3′ and

has a length of 14 to 32 nucleotides,

wherein the CpG is non-methylated CpG without a modified phosphate backbone,

wherein X₁X₂ is AA, AT, GA, or GT, and may comprise a modified phosphate backbone,

wherein X₃X₄ is TT, AT, AC, TA, TC, or CG, and may comprise a modified phosphate backbone, and

the oligonucleotide comprises at least one modified phosphate backbone at a site other than the parts represented by 5′

-X₁X₂CpGX₃X₄-3′

; and

wherein the oligonucleotide comprises one of the following sequences;

7. The oligonucleotide as claimed in claim 3, wherein the oligonucleotide comprises a sequence selected from Sequence Nos. 12, 17, 78, 43, 60, 67, 102, 103, 112 and 116.

4. An oligonucleotide which comprises two to four sequences each represented by 5′

-X₁X₂CpGX₃X₄-3′ and

has a length of 14 to 32 nucleotides,

wherein the CpG is non-methylated CpG without a modified phosphate backbone,

wherein X₁X₂ is AA, AT, GA, or GT, and may comprise a modified phosphate backbone,

wherein X₃X₄ is TT, AT, AC, TA, TC, or CG, and may comprise a modified phosphate backbone,

wherein the oligonucleotide comprises at least one modified phosphate backbone at a site other than the parts represented by 5′

-X₁X₂CpGX₃X₄-3′

, and

wherein the oligonucleotide comprises a sequence represented as -(G)_m-, wherein m is an integer of 1 to 6, at 3′

end side of a CpG motif, the CpG motif being 5′

-X₁X₂CpGX₃X₄-3′

.

5. An oligonucleotide which comprises two to four sequences each represented by 5′

-X₁X₂CpGX₃X₄-3′ and

has a length of 14 to 32 nucleotides,

wherein the CpG is non-methylated CpG without a modified phosphate backbone,

wherein X₁X₂ is AA, AT, GA, or GT, and may comprise a modified phosphate backbone,

wherein X₃X₄ is TT, AT, AC, TA, TC, or CG, and may comprise a modified phosphate backbone,

wherein the oligonucleotide comprises at least one modified phosphate backbone at a site other than the parts represented by 5′

-X₁X₂CpGX₃X₄-3′

, and

wherein the oligonucleotide comprises a sequence represented as TC, TA, TG, CC or CC at 5′

end side of a CpG motif, the CpG motif being the sequence of 5′

-X₁X₂CpGX₃X₄-3′

.

6. An oligonucleotide which comprises two to four sequences each represented by 5′

-X₁X₂CpGX₃X₄-3′ and

has a length of 14 to 32 nucleotides,

wherein the CpG is non-methylated CpG without a modified phosphate backbone,

wherein X₁X₂ is AA, AT, GA, or GT, and may comprise a modified phosphate backbone,

wherein X₃X₄ is TT, AT, AC, TA, TC, or CG, and may comprise a modified phosphate backbone,

wherein the oligonucleotide comprises at least one modified phosphate backbone at a site other than the parts represented by 5′

-X₁X₂CpGX₃X₄-3′

, and

wherein the oligonucleotide comprises at least a first CpG motif and a second CpG motif, the first and the second CpG motifs being 5′

-X₁X₂CpGX₃X₄-3′

,

wherein the first CpG motif and the second CpG motif are bonded directly with no sequence between them, or

wherein the oligonucleotide comprises a sequence represented as -(T)_n-, wherein n is an integer of 1 to 3, TA or TC between the first CpG motif and the second CpG motif.