Composition for targeting dendritic cells

Abstract

The present invention relates to a composition for targeting dendritic cells. In particular, the present invention relates to a composition comprising: a) one or more antigens; b) an anti-DC-SIGN immunoglobulin single variable domain; and c) a carrier which carries a) and b). The invention further relates to formulations, compositions and devices comprising such anti-DC-SIGN molecules and their use as a medicament and in the treatment of cancer, suitably melanoma.

What is claimed is: 
     
         1 . A composition comprising:
 a) one or more antigens;   b) an anti-DC-SIGN immunoglobulin single variable domain; and   c) a carrier which carries a) and b).   
     
     
         2 . The composition according to  claim 1  further comprising d) an immunomodulatory factor. 
     
     
         3 . The composition according to  claim 1  or  2  wherein a) one or more antigen is derived from membrane vesicles (MVs). 
     
     
         4 . The composition according to  claim 3  wherein the membrane vesicles comprise membrane-associated antigens. 
     
     
         5 . The composition according to  claim 4  wherein the membrane-associated antigens are tumour antigens. 
     
     
         6 . The composition according to  claim 5  wherein the membrane-associated antigens comprise tumour antigens selected from the group consisting of melanoma differentiation antigens tyrosinase, gp100 and MART-1, and the cancer testis antigens MAGE-A3, MAGE A-10, BAGE, GAGE and RAGE. 
     
     
         7 . The composition according to any of  claims 1  to  6  wherein the membrane vesicles are derived from tumour cells, melanoma cells, or MM200 melanoma cells. 
     
     
         8 . The composition according to any of  claims 1  to  7  wherein the anti-DC-SIGN immunoglobulin single variable domain is a heavy chain dAb fragment. 
     
     
         9 . The composition according to  claim 8  wherein the anti-DC-SIGN immunoglobulin single variable domain is a V H  dAb fragment. 
     
     
         10 . The composition according to any of  claims 1  to  9  wherein the anti-DC-SIGN immunoglobulin single variable domain comprises SEQ ID NO: 1 or SEQ ID NO: 3 (DMS5000). 
     
     
         11 . The composition according to any of  claims 1  to  10  wherein the anti-DC-SIGN immunoglobulin single variable domain further comprises a polyhistidine C-terminal tail. 
     
     
         12 . The composition according to any of  claims 1  to  11  wherein the carrier is a liposome. 
     
     
         13 . The composition according to  claim 12  wherein the liposome comprises liposomal constituents. 
     
     
         14 . The composition according to  claim 13  wherein the liposomal constituents comprise the chelator lipid 3(nitrilotriacetic acid)-ditetradecylamine(3NTA-DTDA). 
     
     
         15 . The composition according to any of  claim 13  or  14  wherein the liposomal constituents comprise nickel sulphate (NiSO 4 ). 
     
     
         16 . The composition according to any of  claims 13  to  15  wherein the liposomal constituents comprise the lipid α-palmitoyl-β-oleoyl-phosphatidylcholine (POPC)). 
     
     
         17 . The composition according to any of  claims 2  to  16  wherein the immunomodulatory factor is the cytokine interferon gamma (IFN-gamma). 
     
     
         18 . The composition according to any of  claims 1  to  17  wherein the composition is a vaccine composition. 
     
     
         19 . A method for making a composition according to any of  claims 1  to  18  comprising:
 i) preparing membrane vesicles; 
 ii) preparing the liposomal constituent; 
 iii) combining the membrane vesicles and the liposomal constituent with the immunomodulatory factor; and 
 iv) adding the anti-DC-SIGN immunoglobulin single variable domain. 
 
     
     
         20 . The method according to  claim 19  wherein the preparation of membrane vesicles is by propagating tumour cells, sonicating and preparing membrane pellets by centrifugation and resuspension in PBS. 
     
     
         21 . The method according to  claim 19  or  20  wherein the liposomal constituent is prepared by mixing POPC and Ni-3NTA-DTDA. 
     
     
         22 . The method according to any of  claims 19  to  21  further comprising supplementing with nickel. 
     
     
         23 . The composition according to any of  claims 1  to  18  for use as a medicament. 
     
     
         24 . The composition according to any of  claims 1  to  18  for use in the treatment of cancer. 
     
     
         25 . The composition according to  claim 24  wherein the cancer is melanoma. 
     
     
         26 . The composition according to any of  claims 23  to  25  for intravenous administration. 
     
     
         27 . A method for treating a tumour in a subject comprising administering a composition as claimed in any of  claims 1  to  18 . 
     
     
         28 . A composition or method as claimed in any of the preceding claims wherein the anti-DC-SIGN immunoglobulin single variable domain is one which has the same binding specificity as an anti-DC-SIGN immunoglobulin single variable domain having the amino acid sequence as set out in SEQ ID NO: 1 or 3. 
     
     
         29 . A composition or method as claimed in any of the preceding claims wherein the anti-DC-SIGN immunoglobulin single variable domain is one which has the same CDR sequences as the amino acid sequence set out in SEQ ID NO: 1 or 3.