P
USRE45196EExpiredUtilityPatentIndex 45

Compositions comprising structurally stable conjugate molecules

Assignee: TUCK STEPHEN FPriority: Mar 4, 2005Filed: May 18, 2012Granted: Oct 14, 2014
Est. expiryMar 4, 2025(expired)· nominal 20-yr term from priority
Inventors:TUCK STEPHEN FRODRIGUEZ ROBERTO
A61K 2039/6006A61P 37/02A61K 48/00A61K 2039/6025A61K 2039/55561A61K 47/646A61K 39/35A61K 39/385A61K 39/36Y02A50/30
45
PatentIndex Score
1
Cited by
164
References
38
Claims

Abstract

The present invention provides compositions comprising conjugate molecules that are structurally stable at a temperature of between about 2 degrees C. and 8 degrees C. In some examples, a conjugate molecule comprises an antigen, such as an allergen. In some examples, a conjugate molecule comprises the Ragweed antigen Amb a 1. The present invention provides methods for making and using such compositions. Provided herein are methods for modulating an immune response in an individual comprising administration of a composition comprising a structurally stable conjugate molecule as described herein.

Claims

exact text as granted — not AI-modified
What is claimed as new and desired to be protected by Letters Patent of the United States is: 
     
       1. A composition comprising a structurally stable conjugate molecule, wherein the conjugate molecule comprises a conjugate partner and a polynucleotide comprising an immunostimulatory sequence (ISS), wherein the ISS comprises a 5′-CG-3′ sequence, is greater than about 6 bases or base pairs in length, and is less than about 200 bases or base pairs in length, and wherein the composition further comprises a component capable of maintaining the pH of the composition in the range of about 6.0 to about 9.0, wherein the component is Histidine or Phosphate, and wherein the Phosphate, if present, is present in the composition at a concentration of between about 20 mM and about 50 mM. 
     
     
       2. The composition of  claim 1  wherein the composition comprises more than about 70% of said conjugate molecule in non-aggregate form at a temperature of between about 2 degrees C. and about 8 degrees C. as measured by Right Angle Light Scatter (RALS). 
     
     
       3. The composition of  claim 1  wherein the component capable of maintaining the pH is Histidine. 
     
     
       4. The composition of  claim 3  wherein Histidine is present in the composition at a concentration of between about 1 mM and about 50 mM. 
     
     
       5. The composition of  claim 4  wherein Histidine is present in the composition at a concentration of between about 5 mM and about 20 mM. 
     
     
       6. The composition of  claim 1  wherein the component capable of maintaining the pH is Phosphate. 
     
     
       7. The composition of  claim 1  wherein the pH of the composition is in the range of about 7.0 to about 8.0. 
     
     
       8. The composition of  claim 1  wherein the pH of the composition is in the range of about 7.5 to about 8.0. 
     
     
       9. The composition of  claim 1  wherein the ISS comprises the hexamer motif AACGTT. 
     
     
       10. The composition of  claim 1  wherein the ISS comprises the motif
   GACGCTCC; 
   GACGTCCC; 
   GACGTTCC; 
   GACGCCCC; 
   AGCGTTCC; 
   AGCGCTCC; 
   AGCGTCCC; 
   AGCGCCCC; 
   AACGTCCC; 
   AACGCCCC; 
   AACGTTCC; 
   AACGCTCC; 
   GGCGTTCC; 
   GGCGCTCC; 
   GGCGTCCC; 
   GGCGCCCC; 
   GACGCTCG; 
   GACGTCCG; 
   GACGCCCG; 
   GACGTTCG; 
   AGCGCTCG; 
   AGCGTTCG; 
   AGCGTCCG; 
   AGCGCCCG; 
   AACGTCCG; 
   AACGCCCG; 
   AACGTTCG; 
   AACGCTCG; 
   GGCGTTCG; 
   GGCGCTCG; 
   GGCGTCCG; 
   GGCGCCCG; 
   GACGCT; 
   GACGTC; 
   GACGTT; 
   GACGCC; 
   GACGCU; 
   GACGUC; 
   GACGUU; 
   GACGUT; 
   GACGTU; 
   AGCGTT; 
   AGCGCT; 
   AGCGTC; 
   AGCGCC; 
   AGCGUU; 
   AGCGCU; 
   AGCGUC; 
   AGCGUT; 
   AGCGTU; 
   AACGTC; 
   AACGCC; 
   AACGTT; 
   AACGCT; 
   AACGUC; 
   AACGUU; 
   AACGCU; 
   AACGUT; 
   AACGTU; 
   GGCGTT; 
   GGCGCT; 
   GGCGTC; 
   GGCGCC; 
   GGCGUU; 
   GGCGCU; 
   GGCGUC; 
   GGCGUT; 
 or
   GGCGTU. 
 
 
     
     
       11. The composition of  claim 1  wherein the ISS comprises the sequence 
       
         
           
                 
                 
               
                   5′-TGACTGTGAACGTTCGAGATGA-3′; 
                   (SEQ ID NO: 1) 
                 
                     
                 
                   5′-TGACCGTGAACGTTCGAGATGA-3′; 
                   (SEQ ID NO: 2) 
                 
                     
                 
                   5′-TCATCTCGAACGTTCCACAGTCA-3′; 
                   (SEQ ID NO: 3) 
                 
                     
                 
                   5′-TGACTGTGAACGTTCCAGATGA-3′; 
                   (SEQ ID NO: 4) 
                 
                     
                 
                   5′-TCCATAACGTTCGCCTAACGTTCGTC-3′; 
                   (SEQ ID NO: 5) 
                 
                     
                 
                   5′-TGACTGTGAABGTTCCAGATGA-3′; 
                   (SEQ ID NO: 6) 
                 
                     
                 
                   5′-TGACTGTGAABGTTCGAGATGA-3′; 
                   (SEQ ID NO: 7) 
                 
                   or 
                     
                 
                     
                 
                   5′-TGACTGTGAABGTTBGAGATGA-3′, 
                   (SEQ ID NO: 8) 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
       where B is 5-bromocytosine. 
     
     
       12. The composition of  claim 9  wherein the ISS comprises
   5′-TGACTGTGAACGTTCGAGATGA-3′.  (SEQ ID NO:1)
 
 
     
     
       13. The composition of  claim 1  wherein the conjugate partner is an antigen. 
     
     
       14. The composition of  claim 13  wherein the antigen is an allergen. 
     
     
       15. The composition of claim  13  14 wherein the allergen is selected from the group consisting of Crustacea allergens, insect allergens, mammalian allergens, mollusks allergens, plant allergens, and fungal allergens. 
     
     
       16. The composition of  claim 15  wherein the allergen is the plant allergen Ragweed antigen Amb a 1. 
     
     
       17. The composition of  claim 1  further comprising an amino acid selected from the group consisting of Histidine, Glycine, Isoleucine, Leucine, Proline and Alanine. 
     
     
       18. The composition of  claim 17  wherein the amino acid is Glycine. 
     
     
       19. The composition of  claim 18  wherein the Glycine is present in the composition at a concentration of between about 230 mM and about 285 mM. 
     
     
       20. The composition of  claim 1  further comprising a carbohydrate selected from the group consisting of Lactose, Sucrose, Mannose, Maltose, Sorbitol, and Glucose. 
     
     
       21. The composition of  claim 20  wherein the carbohydrate is Sucrose. 
     
     
       22. The composition of  claim 21  wherein the Sucrose is present in the composition at a concentration of between about 1% and 10%. 
     
     
       23. The composition of  claim 20  wherein the carbohydrate is Sorbitol. 
     
     
       24. The composition of  claim 23  wherein the Sorbitol is present in the composition at a concentration of between about 3% and about 5%. 
     
     
       25. A composition comprising a conjugate molecule, wherein the conjugate molecule comprises a conjugate partner and a polynucleotide comprising an immunostimulatory sequence (ISS), wherein the ISS comprises a 5′-CG-3′ sequence, is greater than about 6 bases or base pairs in length, and is less than about 200 bases or base pairs in length, and wherein the composition further comprises Histidine at a concentration in the range of about 1 mM to about 50 mM; and Glycine at a concentration in the range of about 50 mM to about 300 mM, wherein said composition has a pH in the range of about 6.0 to about 9.0. 
     
     
       26. The composition of  claim 25  further comprising Sorbitol in the range of about 1 to 10%. 
     
     
       27. The composition of  claim 25  further comprising Sucrose at a concentration of about 200 mM to about 250 mM. 
     
     
       28. The composition of  claim 25  wherein said composition has a pH in the range of about 7.0 to about 8.0. 
     
     
       29. The composition of  claim 28  comprising 5 mM Histidine and 285 mM Glycine. 
     
     
       30. The composition of  claim 28  comprising 20 mM Histidine and 270 mM Glycine. 
     
     
       31. The composition of  claim 28  comprising 20 mM Histidine, 50 mM Glycine, and 3.8% Sorbitol. 
     
     
       32. The composition of  claim 28  comprising 20 mM Histidine, 50 mM Glycine, and 210 mM Sucrose. 
     
     
       33. The composition of  claim 25  wherein the conjugate partner is an allergen. 
     
     
       34. The composition of  claim 33  wherein the allergen is a plant allergen. 
     
     
       35. The composition of  claim 34  wherein the plant allergen is Ragweed antigen Amb a1. 
     
     
       36. A lyophilized composition produced by lyophilizing the composition of  claim 20  under suitable conditions. 
     
     
       37. A lyophilized composition comprising the composition of  claim 26  or  27 . 
     
     
       38. The composition of claim 1 or claim 25, wherein the polynucleotide is single-stranded DNA.

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