Synthetic hla binding peptide analogues and uses thereof

Abstract

The present invention is directed to a synthetic peptide comprising a sequence of amino acids containing at least a segment that is an analogue of a native peptide that specifically binds to HLA A0201 or HLA A0301 molecules on a cell characteristic of a pathophysiologic state in a mammal. The synthetic peptide may be derived form native peptides comprising a breakpoint region of the WT1 protein.

1 . A synthetic WT-1 peptide comprising a sequence of amino acids selected from the group consisting of YMFPNAPYL (SEQ ID NO: 18), YLGEQQYSV (SEQ ID NO: 20), YLLPAVPSL (SEQ ID NO: 22), YLGATLKGV (SEQ ID NO: 24), YLNALLPAV (SEQ ID NO: 26), GLRRGIQDV (SEQ ID NO: 28), KLYFKLSHL (SEQ ID NO: 30), ALLLRTPYV (SEQ ID NO: 32), YMTWNQMNL (SEQ ID NO: 34), NMYQRNMTK (SEQ ID NO: 36), NMHQRVMTK (SEQ ID NO: 37), NMYQRVMTK (SEQ ID NO: 38), QMYLGATLK (SEQ ID NO: 40), QMNLGVTLK (SEQ ID NO: 41), QMYLGVTLK (SEQ ID NO: 42), FMYAYPGCNK (SEQ ID NO: 44), FMCAYPFCNK (SEQ ID NO: 45), FMYAYPFCNK (SEQ ID NO: 46), KLYHLQMHSR (SEQ ID NO: 48), KLSHLQMHSK (SEQ ID NO: 49), and KLYHLQMHSK (SEQ ID NO: 50), that is an analogue peptide of a native WT-1 peptide selected from the group consisting of RMFPNAPYL (SEQ ID NO: 17), SLGEQQYSV (SEQ ID NO: 19), ALLPAVPSL (SEQ ID NO: 21), NLGATLKGV (SEQ ID NO: 23), DLNALLPAV (SEQ ID NO: 25), GVFRGIQDV (SEQ ID NO: 27), KRYFKLSHL (SEQ ID NO: 29), ALLLRTPYS (SEQ ID NO: 31), CMTWNQMNL (SEQ ID NO: 33), NMHQRNMTK (SEQ ID NO: 35), QMNLGATLK (SEQ ID NO: 39), FMCAYPGCNK (SEQ ID NO: 43), and KLSHLQMHSR (SEQ ID NO: 47), wherein said analogue specifically binds to HLA A0201 or HLA A0301 molecules on a human cell. 
     
     
         2 . The synthetic WT-1 peptide of  claim 1 , wherein said analogue peptide is a degradation product of said synthetic WT-1 peptide. 
     
     
         3 . The synthetic WT-1 peptide of  claim 1 , further comprising: an immunogenic carrier linked thereto. 
     
     
         4 . The synthetic WT-1 peptide of  claim 3 , wherein said immunogenic carrier is a carrier protein, a carrier peptide or an antigen-presenting cell. 
     
     
         5 . The synthetic WT-1 peptide of  claim 4 , wherein said carrier protein or carrier peptide is keyhole limpet hemocyanin, an albumin or a polyamino acid. 
     
     
         6 . The synthetic WT-1 peptide of  claim 4 , wherein said antigen presenting cell is a dendritic cell. 
     
     
         7 . The synthetic WT-1 peptide of  claim 1 , wherein a total number of amino acids in said analogue peptide is about 70% to about 130% of a total number of amino acids in said native WT-1 peptide. 
     
     
         8 . The synthetic WT-1 peptide of  claim 8 , wherein said analogue peptide has about 8 to about 12 amino acids. 
     
     
         9 .- 11 . (canceled) 
     
     
         12 . The synthetic WT-1 peptide of  claim 1 , wherein said human cell is characteristic of a pathophysiologic state. 
     
     
         13 . The synthetic WT-1 peptide of  claim 12 , wherein said pathophysiologic state is a cancer selected from the group consisting of a leukemia, a breast cancer, a lymphoma, a mesothelioma, a lung cancer, a testicular cancer, and an ovarian cancer. 
     
     
         14 . The synthetic WT-1 peptide of  claim 13 , wherein said leukemia is a chronic myelogenous leukemia. 
     
     
         15 . A synthetic WT-1 peptide having an amino acid sequence selected from the group consisting of NMYQRNMTK (SEQ ID NO: 36), NMHQRVMTK (SEQ ID NO: 37), NMYQRVMTK (SEQ ID NO: 38), QMYLGATLK (SEQ ID NO: 40), 25 QMNLGVTLK (SEQ ID NO: 41), QMYLGVTLK (SEQ ID NO: 42), FMYAYPGCNK (SEQ ID NO: 44), FMCAYPFCNK (SEQ ID NO: 45), FMYAYPFCNK (SEQ ID NO: 46), KLYHLQMHSR (SEQ ID NO: 48), KLSHLQMHSK (SEQ ID NO: 49), or KLYHLQMHSK (SEQ ID NO: 50). 
     
     
         16 . A method of inducing formation and proliferation of human cytotoxic T cells that recognize a cancer cell, wherein said cancer cell presents the native WT-1 peptide of  claim 1  on a major histocompatibility complex (MHC) class I molecule thereof, said method comprising contacting human immune cells in vivo or ex vivo with the synthetic WT-1 peptide of  claim 1  or DNA encoding said synthetic WT-1 peptide, whereby said synthetic WT-1 peptide or DNA encoding said synthetic peptide induces formation and proliferation of said human cytotoxic T cells. 
     
     
         17 . The method of  claim 16 , whereby said human cytotoxic T cells are capable of mounting a heteroclitic immune response against said cancer cell. 
     
     
         18 . The method of  claim 16 , wherein the step of contacting said human immune cells are is performed in vivo in an individual having a cancer. 
     
     
         19 . A method of treating a subject having a cancer, wherein a malignant cell of said cancer presents the native WT-1 peptide of  claim 16  on a major histocompatibility complex (MHC) class I molecule thereof, comprising
 a. inducing in vivo in a donor or ex vivo using immune cells from a donor formation and proliferation of human cytotoxic T cells that recognize said malignant cell by the method of  claim 16 ; and 
 b. infusing said human cytotoxic T cells into said subject, 
 
       thereby treating a subject having a cancer. 
     
     
         20 .- 21 . (canceled) 
     
     
         22 . The method of  claim 16 , whereby said DNA molecule is in a vector or an antigen presenting cell. 
     
     
         23 . The method of  claim 16 , wherein said human immune cells are peripheral blood mononuclear cells, bone marrow cells, dendritic cells, or macrophages. 
     
     
         24 . The method of  claim 16 , wherein said cancer cell is selected from the group consisting of a leukemia cell, a breast cancer cell, a lymphoma cell, a mesothelioma cell, a lung cancer cell, a testicular cancer cell, and an ovarian cancer cell. 
     
     
         25 .- 29 . (canceled) 
     
     
         30 . A pharmaceutical composition, comprising: a therapeutically effective amount of the synthetic WT-1 peptide of  claim 1  or a DNA encoding said synthetic WT-1 peptide; and a pharmaceutically acceptable carrier. 
     
     
         31 . The pharmaceutical composition of  claim 30 , wherein said DNA is in a vector or an antigen presenting cell. 
     
     
         32 .- 33 . (canceled) 
     
     
         34 . The pharmaceutical composition of  claim 30 , further comprising an adjuvant, a diluent or a combination thereof. 
     
     
         35 . The pharmaceutical composition of  claim 34 , wherein said adjuvant is a protein, a peptide or an antigen presenting cell linked to said synthetic peptide. 
     
     
         36 . The pharmaceutical composition of  claim 35 , wherein said protein or peptide is keyhole limpet hemocyanin, an albumin or a polyamino acid. 
     
     
         37 . The pharmaceutical composition of  claim 35 , wherein said antigen presenting cell is a dendritic cell. 
     
     
         38 .- 42 . (canceled) 
     
     
         43 . The method of  claim 16  wherein said synthetic WT-1 peptide further comprises an adjuvant.