Polypeptides and polynucleotides for enhancing immune reactivity to her-2 protein

Abstract

Compositions for stimulating the immune system and for treating malignancies associated with overexpression of the HER-2 protein are provided. Such compositions include immunogenic epitopes of the HER-2 proteins and chimeric and multivalent peptides which comprise such epitopes. The present invention also relates to polynucleotides which encode the chimeric peptides. Also provided are pharmaceutical compositions comprising such immunogenic compositions. Methods for stimulating an immune response to HER-2 protein are provided. Methods for treating breast cancer, ovarian cancer, prostate cancer, colon cancer and lung cancer are provided.

1 . A chimeric peptide for stimulating an immune response to HER-2 protein comprising a HER-2 B cell epitope, a T helper (Th) epitope, and a linker joining the HER-2 B cell epitope to the Th epitope, wherein:
 the HER-2 B cell epitope consists of a sequence selected from the group consisting of:   
       
         
           
                 
                 
               
                     
                   SEQ ID NO: 1 
                 
                     
                   TGTDMKLRLPASPETHLDM,; 
                 
                     
                     
                 
                     
                   SEQ ID NO: 2 
                 
                     
                   AVLDNGDPLNNTTPVTGASPGG,; 
                 
                     
                     
                 
                     
                   SEQ ID NO: 3 
                 
                     
                   LWKDIFHKNNQLALTLIDTNRS,; 
                 
                     
                     
                 
                     
                   SEQ ID NO: 4 
                 
                     
                   TLIDTNRSRACHPCSPMCKGSRCWGESSEDCQSLT,; 
                 
                     
                     
                 
                     
                   SEQ ID NO: 7 
                 
                     
                   PESFDGDPASNTAPLQPE,; 
                 
                     
                     
                 
                     
                   SEQ ID NO: 8 
                 
                     
                   LYISAWPDSLPDLSVFQNLQ,; 
                 
                     
                     
                 
                     
                   SEQ ID NO: 9 
                 
                     
                   LFRNPHQALLHTANRPEDE,; 
                 
                     
                     
                 
                     
                   SEQ ID NO: 10 
                 
                     
                   CLPCHPECQPQNGSVTCFGPEADQCVACAHYKDP,; 
                 
                     
                   and 
                 
                     
                     
                 
                     
                   SEQ ID NO: 12 
                 
                     
                   INGTHSCVDLDDKGCPAEQRAS,; 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
         the Th epitope comprises a sequence selected from the group consisting of: 
       
       
         
           
                 
                 
               
                     
                   SEQ ID NO: 13 
                 
                     
                   NSVDDALINSTIYSYFPSV,; 
                 
                     
                     
                 
                     
                   SEQ ID NO: 14 
                 
                     
                   PGINGKAIHLVNNQSSE,; 
                 
                     
                     
                 
                     
                   SEQ ID NO: 15 
                 
                     
                   QYIKANSKFIGITEL,; 
                 
                     
                     
                 
                     
                   SEQ ID NO: 16 
                 
                     
                   FNNFTVSFWLRVPKVSASHLE,; 
                 
                     
                     
                 
                     
                   SEQ ID NO: 17 
                 
                     
                   LSEIKGVIVHRLEGV,; 
                 
                     
                     
                 
                     
                   SEQ ID NO: 18 
                 
                     
                   FFLLTRILTIPQSLN,; 
                 
                     
                   and 
                 
                     
                     
                 
                     
                   SEQ ID NO: 19 
                 
                     
                   TCGVGVRVRSRVNAANKKPE,; 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
         and 
         the linker comprises a sequence that is from 1 to 15 amino acids in length. 
       
     
     
         2 . The chimeric peptide of  claim 1  wherein the Th epitope comprises NSVDDALINSTIYSYFPSV, SEQ ID NO: 13. 
     
     
         3 . The chimeric peptide of  claim 1  wherein the Th epitope comprises PGINGKAIHLVNNQSSE, SEQ ID NO: 14. 
     
     
         4 . The chimeric peptide of  claim 1  wherein the Th epitope comprises QYIKANSKFIGITEL, SEQ ID NO: 15. 
     
     
         5 . The chimeric peptide of  claim 1  wherein the Th epitope comprises FNNFTVSFWLRVPKVSASHLE, SEQ ID NO: 16. 
     
     
         6 . The chimeric peptide of  claim 1  wherein the Th epitope comprises LSEIKGVIVHRLEGV, SEQ ID NO: 17. 
     
     
         7 . The chimeric peptide of  claim 1  wherein the Th epitope comprises FFLLTRILTIPQSLN, SEQ ID NO: 18. 
     
     
         8 . The chimeric peptide of  claim 1  wherein the Th epitope comprises TCGVGVRVRSRVNAANKKPE, SEQ ID NO: 19. 
     
     
         9 . The chimeric peptide of  claim 1  wherein the linker comprises a sequence that is from 2 to 15 amino acids in length. 
     
     
         10 . The chimeric peptide of  claim 9  wherein the linker comprises GPSL, SEQ ID NO: 20. 
     
     
         11 . The chimeric peptide of  claim 1  further comprising a second HER-2 B cell epitope comprising a sequence selected from the group consisting of: 
       
         
           
                 
                 
               
                     
                   SEQ ID NO: 1 
                 
                     
                   TGTDMKLRLPASPETHLDM,; 
                 
                     
                     
                 
                     
                   SEQ ID NO: 2 
                 
                     
                   AVLDNGDPLNNTTPVTGASPGG,; 
                 
                     
                     
                 
                     
                   SEQ ID NO: 3 
                 
                     
                   LWKDIFHKNNQLALTLIDTNRS,; 
                 
                     
                     
                 
                     
                   SEQ ID NO: 4 
                 
                     
                   TLIDTNRSRACHPCSPMCKGSRCWGESSEDCQSLT,; 
                 
                     
                     
                 
                     
                   SEQ ID NO: 5 
                 
                     
                   ALVTYNTDTFESMPNPEGRYT,; 
                 
                     
                     
                 
                     
                   SEQ ID NO: 6 
                 
                     
                   PLHNQEVTAEDGTQRAEKCSKPCA,; 
                 
                     
                     
                 
                     
                   SEQ ID NO: 7 
                 
                     
                   PESFDGDPASNTAPLQPE,; 
                 
                     
                     
                 
                     
                   SEQ ID NO: 8 
                 
                     
                   LYISAWPDSLPDLSVFQNLQ,; 
                 
                     
                     
                 
                     
                   SEQ ID NO: 9 
                 
                     
                   LFRNPHQALLHTANRPEDE,; 
                 
                     
                     
                 
                     
                   SEQ ID NO: 10 
                 
                     
                   CLPCHPECQPQNGSVTCFGPEADQCVACAHYKDP,; 
                 
                     
                     
                 
                     
                   SEQ ID NO: 11 
                 
                     
                   KPDLSYMPIWKFPDEEGA,; 
                 
                     
                   and 
                 
                     
                     
                 
                     
                   SEQ ID NO: 12 
                 
                     
                   INGTHSCVDLDDKGCPAEQRAS,. 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         12 . The chimeric peptide of  claim 11  further comprising a second linker joining the first HER-2 B cell epitope to the second HER-2 B cell epitope. 
     
     
         13 . The chimeric peptide of  claim 12  wherein the second linker comprises a sequence that is from 1 to 15 amino acids in length. 
     
     
         14 . The chimeric peptide of  claim 13  wherein the second linker comprises GPSL, SEQ ID NO: 20. 
     
     
         15 . A method of stimulating an immune response in a subject comprising administering a pharmaceutical composition to the subject, the pharmaceutical composition comprising:
 a chimeric peptide of  claim 1  or  claim 11 ; and   a pharmaceutically acceptable vehicle.   
     
     
         16 . The method of  claim 15  wherein the subject is a human and has one of the following cancers or a predisposition to one of the following cancers: breast cancer, ovarian cancer, lung cancer, prostate cancer, and colon cancer. 
     
     
         17 . The method of  claim 15  wherein the vehicle is biodegradable and is selected from the group consisting of an emulsion comprising a pharmaceutically acceptable oil/water emulsion and a biodegradable microsphere or nanosphere comprising a polylactide-polyglycolic acid polymer. 
     
     
         18 . The method of  claim 17  wherein the oil is squalene or squalene. 
     
     
         19 . The method of  claim 17  wherein the microsphere is from 0.1 to 50 nanometers in diameter and comprises poly ( D, L  lactide-co-glycide). 
     
     
         20 . An immunogenic composition comprising a chimeric peptide of  claim 1  or  claim 11 .