Immunostimulatory properties of oligonucleotide-based compounds comprising modified immunostimulatory dinucleotides

Abstract

The invention relates to the therapeutic use of oligonucleotides as immunostimulatory agents in immunotherapy applications. More particularly, the invention provides immunostimulatory oligonucleotides for use in methods for generating an immune response or for treating a patient in need of immunostimulation. The immunostimulatory oligonucleotides of the invention preferably comprise novel purines. The immunostimulatory oligonucleotides according to the invention further comprise at least two oligonucleotides linked at their 3′ ends, internucleoside linkages or functionalized nucleobase or sugar to a non-nucleotidic linker, at least one of the oligonucleotides being an immunostimulatory oligonucleotide and having an accessible 5′ end.

1 . A method for therapeutically treating a vertebrate having cancer, an autoimmune disorder, airway inflammation, inflammatory disorders, skin disorders, allergy, asthma or a disease caused by a pathogen, such method comprising administering to the patient an immunostimulatory oligonucleotide having a structure from the group of 5′-TCAGTCG 2 TTAG-X-GATTG 2 CTGACT-5′ (SEQ ID NO: 2); 5′-TCG 2 TCG 2 TTAGA-X-AGATTG 2 CTG 2 CT-5′ (SEQ ID NO: 3); 5′-CAGTCG 2 TTCAG-X-GACTTG 2 CTGAC-5′ (SEQ ID NO: 4); 5′-TCAGTCG 1 TTAG-X-GATTG 1 CTGACT-5′ (SEQ ID NO: 5); 5′-TCG 1 TCG 1 TTAGA-X-AGATTG 1 CTG 1 CT-5′ (SEQ ID NO: 6) and 5′-CAGTCG 1 TTCAG-X-GACTTG 1 CTGAC-5′ (SEQ ID NO: 7); wherein G 1 =2′-deoxy-7-deazaguanosine; G 2 =arabinoguanosine; and X=glycerol linker. 
     
     
         2 . The method according to  claim 1 , wherein the route of administration is selected from parenteral, oral, sublingual, transdermal, topical, intranasal, aerosol, intraocular, intratracheal, intrarectal, vaginal, gene gun, dermal patch, eye drop and mouthwash. 
     
     
         3 . The method according to  claim 1  comprising administering an immunostimulatory oligonucleotide having the structure 5′-TCAGTCG 2 TTAG-X-GATTG 2 CTGACT-5′(SEQ ID NO: 2). 
     
     
         4 . The method according to  claim 1  comprising administering an immunostimulatory oligonucleotide having the structure 5′-TCG 2 TCG 2 TTAGA-X-AGATTG 2 CTG 2 CT-5′(SEQ ID NO: 3). 
     
     
         5 . The method according to  claim 1  comprising administering an immunostimulatory oligonucleotide having the structure 5′-CAGTCG 2 TTCAG-X-GACTTG 2 CTGAC-5′(SEQ ID NO: 4). 
     
     
         6 . The method according to  claim 1  comprising administering an immunostimulatory oligonucleotide having the structure 5′-TCAGTCG 1 TTAG-X-GATTG 1 CTGACT-5′(SEQ ID NO: 5). 
     
     
         7 . The method according to  claim 1  comprising administering an immunostimulatory oligonucleotide having the structure 5′-TCG 1 TCG 1 TTAGA-X-AGATTG 1 CTG 1 CT-5′(SEQ ID NO: 6). 
     
     
         8 . The method according to  claim 1  comprising administering an immunostimulatory oligonucleotide having the structure 5′-CAGTCG 1 TTCAG-X-GACTTG 1 CTGAC-5′(SEQ ID NO: 7). 
     
     
         9 . A method for preventing cancer, an autoimmune disorder, airway inflammation, inflammatory disorders, skin disorders, allergy, asthma or a disease caused by a pathogen in a vertebrate, such method comprising administering to the vertebrate an immunostimulatory oligonucleotide having a structure from the group of 5′-TCAGTCG 2 TTAG-X-GATTG 2 CTGACT-5′ (SEQ ID NO: 2); 5′-TCG 2 TCG 2 TTAGA-X-AGATTG 2 CTG 2 CT-5′ (SEQ ID NO: 3); 5′-CAGTCG 2 TTCAG-X-GACTTG 2 CTGAC-5′ (SEQ ID NO: 4); 5′-TCAGTCG 1 TTAG-X-GATTG 1 CTGACT-5′ (SEQ ID NO: 5); 5′-TCG 1 TCG 1 TTAGA-X-AGATTG 1 CTG 1 CT-5′ (SEQ ID NO: 6) and 5′-CAGTCG 1 TTCAG-X-GACTTG 1 CTGAC-5′ (SEQ ID NO: 7); wherein G 1 =2′-deoxy-7-deazaguanosine; G 2 =arabinoguanosine; and X=glycerol linker. 
     
     
         10 . The method according to  claim 9 , wherein the route of administration is selected from parenteral, oral, sublingual, transdermal, topical, intranasal, aerosol, intraocular, intratracheal, intrarectal, vaginal, gene gun, dermal patch, eye drop and mouthwash. 
     
     
         11 . The method according to  claim 9  comprising administering an immunostimulatory oligonucleotide having the structure 5′-TCAGTCG 2 TTAG-X-GATTG 2 CTGACT-5′(SEQ ID NO: 2). 
     
     
         12 . The method according to  claim 9  comprising administering an immunostimulatory oligonucleotide having the structure 5′-TCG 2 TCG 2 TTAGA-X-AGATTG 2 CTG 2 CT-5′(SEQ ID NO: 3). 
     
     
         13 . The method according to  claim 9  comprising administering an immunostimulatory oligonucleotide having the structure 5′-CAGTCG 2 TTCAG-X-GACTTG 2 CTGAC-5′(SEQ ID NO: 4). 
     
     
         14 . The method according to  claim 9  comprising administering an immunostimulatory oligonucleotide having the structure 5′-TCAGTCG 1 TTAG-X-GATTG 1 CTGACT-5′(SEQ ID NO: 5). 
     
     
         15 . The method according to  claim 9  comprising administering an immunostimulatory oligonucleotide having the structure 5′-TCG 1 TCG 1 TTAGA-X-AGATTG 1 CTG 1 CT-5′(SEQ ID NO: 6). 
     
     
         16 . The method according to  claim 9  comprising administering an immunostimulatory oligonucleotide having the structure 5′-CAGTCG 1 TTCAG-X-GACTTG 1 CTGAC-5′(SEQ ID NO: 7). 
     
     
         17 . The method according to  claim 1 , further comprising administering an antibody, antisense oligonucleotide, protein, antigen, allergen, chemotherapeutic agent or adjuvant. 
     
     
         18 . The method according to  claim 9 , further comprising administering an antibody, antisense oligonucleotide, protein, antigen, allergen, chemotherapeutic agent or adjuvant. 
     
     
         19 . A pharmaceutical composition comprising an immunostimulatory oligonucleotide having a structure from the group of 5′-TCAGTCG 2 T′TAG-X-GA′TTG 2 CTGACT-5′ (SEQ ID NO: 2); 5′-TCG 2 TCG 2 TTAGA-X-AGATTG 2 CTG 2 CT-5′ (SEQ ID NO: 3); 5′-CAGTCG 2 TTCAG-X-GACTTG 2 CTGAC-5′ (SEQ ID NO: 4); 5′-TCAGTCG 1 TTAG-X-GATTG 1 CTGACT-5′ (SEQ ID NO: 5); 5′-TCG 1 TCG 1 TTAGA-X-AGATTG 1 CTG 1 CT-5′ (SEQ ID NO: 6) and 5% CAGTCG 1 TTCAG-X-GACTTG 1 CTGAC-5′ (SEQ ID NO: 7); wherein G 1 =2′-deoxy-7-deazaguanosine; G 2 =arabinoguanosine; and X=glycerol linker, and a physiologically acceptable carrier. 
     
     
         20 . The pharmaceutical composition according to  claim 18 , further comprising administering an antibody, antisense oligonucleotide, protein, antigen, allergen, chemotherapeutic agent or adjuvant.