Amphiphilic oligodeoxynucleotide conjugates as adjuvant enhancers

Abstract

Amphiphilic oligonucleotide conjugates that enhance adjuvant function are disclosed. The conjugates typically include: a lipophilic component, and conjugated thereto (directly or indirectly) an immunomodulating oligonucleotide that, if it were not conjugated to the lipophilic component, would suppress TLR7 and/or TLR8 stimulation. In the presence of albumin, these conjugates significantly enhance adjuvant function, in particular the function of TLR7/8-mediated adjuvants such as an imidazoquinolinamine. The conjugates can be administered, along with an adjuvant compound, to a subject in order to cause and/or enhance an immune response (for instance, to an infectious agent or a cancer antigen) in the subject.

What is claimed is: 
     
         1 . A method, comprising administering to a subject a therapeutically effective amount of an amphiphilic oligonucleotide conjugate comprising:
 a lipophilic component comprising:
 a C14 diacyl lipid, a C18 diacyl lipid, a cholesterol, or 
   
       
         
           
           
               
               
           
         
       
       and
 directly or indirectly conjugated thereto, an immunomodulating oligonucleotide that, if it was not conjugated to the lipophilic component, would suppress TLR7 and/or TLR8 stimulation, the immunomodulating oligonucleotide comprising:
 a poly-oligo(dT) of 10-50 nucleotides (T10-T50), a poly-oligo(dT) of 20-25 nucleotides (T20-T25), or a sequence selected from SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 4. 
 
 
     
     
         2 . The method of  claim 1 , wherein the lipophilic component of the amphiphilic oligonucleotide conjugate is conjugated to the immunomodulating oligonucleotide at its 5′ or 3′ terminal end. 
     
     
         3 . The method of  claim 1 , wherein the immunomodulating oligonucleotide of the amphiphilic oligonucleotide conjugate comprises at least one phosphorothioate bond or at least one other modified (non-naturally occurring) bond. 
     
     
         4 . The method of  claim 1 , wherein the amphiphilic oligonucleotide conjugate further comprises a linker between the lipophilic component and the immunomodulating oligonucleotide. 
     
     
         5 . The method of  claim 1 , wherein the amphiphilic oligonucleotide conjugate comprises: 
       
         
           
           
               
               
           
         
         wherein the value of n is 20; or 
       
       
         
           
           
               
               
           
         
       
     
     
         6 . A method of enhancing an immunostimulatory effect of an imidazoquinolinamine, comprising administering to a cell:
 a therapeutically effective amount of the imidazoquinolinamine; and   an amphiphilic oligonucleotide conjugate comprising:
 a lipophilic component comprising:
 a C14 diacyl lipid, a C18 diacyl lipid, a cholesterol, or 
 
   
       
         
           
           
               
               
           
         
         and
 directly or indirectly conjugated thereto, an immunomodulating oligonucleotide that, if it was not conjugated to the lipophilic component, would suppress TLR7 and/or TLR8 stimulation, the immunomodulating oligonucleotide comprising:
 a poly-oligo(dT) of 10-50 nucleotides (T10-T50), a poly-oligo(dT) of 20-25 nucleotides (T20-T25), or a sequence selected from SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 4. 
 
 
       
     
     
         7 . The method of  claim 6 , wherein the lipophilic component is conjugated to the immunomodulating oligonucleotide at its 5′ or 3′ terminal end. 
     
     
         8 . The method of  claim 6 , wherein the immunomodulating oligonucleotide comprises at least one phosphorothioate bond or at least one other modified (non-naturally occurring) bond. 
     
     
         9 . The method of  claim 6 , further comprising a linker between the lipophilic component and the immunomodulating oligonucleotide. 
     
     
         10 . The method of  claim 6 , comprising: 
       
         
           
           
               
               
           
         
         wherein the value of n is 20; or 
       
       
         
           
           
               
               
           
         
       
     
     
         11 . The method of  claim 6 , wherein the imidazoquinolinamine comprises imiquimod (R-837; IMQ), resiquimod (R-848), or gardiquimod. 
     
     
         12 . The method of  claim 6 , wherein the cell is in vivo in a subject. 
     
     
         13 . A pharmaceutical composition, comprising an amphiphilic oligonucleotide conjugate comprising:
 a lipophilic component comprising:
 a C14 diacyl lipid, a C18 diacyl lipid, a cholesterol, or 
   
       
         
           
           
               
               
           
         
       
       and
 directly or indirectly conjugated thereto, an immunomodulating oligonucleotide that, if it was not conjugated to the lipophilic component, would suppress TLR7 and/or TLR8 stimulation, the immunomodulating oligonucleotide comprising:
 a poly-oligo(dT) of 10-50 nucleotides (T10-T50), a poly-oligo(dT) of 20-25 nucleotides (T20-T25), or a sequence selected from SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 4. 
 
 
     
     
         14 . The pharmaceutical composition of  claim 13 , wherein the lipophilic component of the amphiphilic oligonucleotide conjugate is conjugated to the immunomodulating oligonucleotide at its 5′ or 3′ terminal end. 
     
     
         15 . The pharmaceutical composition of  claim 13 , wherein the immunomodulating oligonucleotide of the amphiphilic oligonucleotide conjugate comprises at least one phosphorothioate bond or at least one other modified (non-naturally occurring) bond. 
     
     
         16 . The pharmaceutical composition of  claim 13 , further comprising a linker between the lipophilic component and the immunomodulating oligonucleotide of the amphiphilic oligonucleotide conjugate. 
     
     
         17 . The pharmaceutical composition of  claim 13 , wherein the amphiphilic oligonucleotide conjugate comprises: 
       
         
           
           
               
               
           
         
         wherein the value of n is 20; or 
       
       
         
           
           
               
               
           
         
       
     
     
         18 . The pharmaceutical composition of  claim 13 , formulated for one or more of:
 administration by a parenteral route;   administration by a transdermal route;   administration by a transmucosal route;   administration using a bioerodible insert; or   in a dosage form appropriate for a route of administration.   
     
     
         19 . The pharmaceutical composition of  claim 18 , which is:
 formulated for administration by a parenteral route of delivery selected from intramuscular, intraperitoneal, intravenous (IV), and subcutaneous injection; or   formulated for administration by a transdermal route of delivery selected from passive delivery, using iontophoresis, and using electroporation; or   formulated for administration by a transmucosal route of delivery selected from nasal, vaginal, rectal, and sublingual.   
     
     
         20 . The pharmaceutical composition of  claim 13 , further comprising one or more of:
 an antigen (such as a viral antigen, a bacterial antigen, a parasite antigen, an allergen, an environmental antigen, or a cancer antigen);   a TLR7- or TLR8-mediated adjuvant (such as a single-stranded RNA, an oligoribonucleotide (ORN), a base analog, or an imidazoquinolinamine);   an adjuvant that is not a TLR7- or TLR8-mediated adjuvant;   a pharmaceutically acceptable diluent;   a preservative;   a solubilizer;   an emulsifier;   an adjuvant; and/or   a carrier.