Xenoantigen-Displaying Anti-Cancer Vaccines and Method of Making

Abstract

Compositions, methods of making, and methods of using, xenoantigen-displaying anticancer vaccines are described. In another broad aspect, there is provided herein a method of synthesizing an alkynefunctionalized composition of claim 1 , comprising: deprotecting an ester comprising a Fmoc moiety to form a free acid; coupling the free acid of step (a) with an amine; and, removing the Fmoc moiety, and coupling the remaining moiety with palmitic acid to yield an alkyne-functionalized composition.

1 . A composition comprising a first lipid (lipid a ) moiety and an alkyne amide moiety having a Formula IV: 
       
         
           
           
               
               
           
         
       
     
     
         2 . The composition of  claim 1 , wherein the first lipid (lipid a ) moiety comprises a Toll-like receptor (TLR) agonist ligand selected from one or more of: TLR2, TLR1, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, TLR10, TLR11, TLR12, TLR13, TLR14, TLR15 and TLR16. 
     
     
         3 . The composition of  claim 1 , wherein the lipid moiety comprises:
 dipalmitoyl-S-glyceryl-cys-(Pam 2 Cys-);   tripalmitoyl-S-glyceryl-cys-(Pam 3 Cys-);   dipalmitoyl-S-glyceryl-cys-ser-lys-lys-lys-lys (Pam 2 Cys-Ser-(Lys) 4 ) [SEQ ID NO: 2];   tripalmitoyl-S-glyceryl-cys-ser-lys-lys-lys-lys (Pam 3 Cys-Ser-(Lys) 4 ) [SEQ ID NO: 3]; or   MALP-2 dipalmitoyl-S-glyceryl-cys-gly-asn-asn-asp-glu-ser-asn-ile-ser-phe-lys-glu-lys (Pam 2 CGNNDESNISFKEK)] [SEQ ID NO: 4].   
     
     
         4 . The composition of  claim 1 , wherein Formula IV comprises an alkyne-functionalized Pam 3 Cys amide composition (6) comprising: 
       
         
           
           
               
               
           
         
       
       wherein Pam is a dipalmitoyl-S-glyceryl-moiety. 
     
     
         5 . A method of synthesizing an alkyne-functionalized composition of  claim 1 , comprising:
 a) deprotecting an ester comprising a Fmoc moiety to form a free acid;   b) coupling the free acid of step (a) with an amine; and,   c) removing the Fmoc moiety, and coupling the remaining moiety with palmitic acid to yield an alkyne-functionalized composition.   
     
     
         6 . A method of synthesizing an alkyne-functionalized Pam 3 Cys amide composition (6) of  claim 4 , comprising:
 a) deprotecting 0-palmitoylated Fmoc L-cystine tert-butyl ester (4) to form a free acid;   b) coupling the free acid of step (a) with propargyl amine in presence of benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (PyBOP), 1-hydroxy-benzotriazole (HOBt) and N,N-diisopropylethylamine (DIPEA) to yield composition (5); and,   c) removing a Fmoc group of composition (5) by treatment with a mixture of acetonitrile-dichloromethane-diethyl amine,   followed by subsequent palmitoylation by coupling with palmitic acid, PyBOP, HOBt and DIPEA to yield the alkyne-functionalized Pam 3 Cys amide composition (6).   
     
     
         7 . A lipidated glycopeptide composition comprising: a first lipid (lipid a ) moiety, a first linker (linker a ) moiety, and an antigen moiety, having a Formula II: 
       
         
           
           
               
               
           
         
       
     
     
         8 . (canceled) 
     
     
         9 . The composition of  claim 7 , wherein the first lipid (lipid a ) moiety comprises:
 dipalmitoyl-S-glyceryl-cys-(Pam 2 Cys-);   tripalmitoyl-S-glyceryl-cys-(Pam 3 Cys-);   dipalmitoyl-S-glyceryl-cys-ser-lys-lys-lys-lys (Pam 2 Cys-Ser-(Lys) 4 ) [SEQ ID NO: 2];   tripalmitoyl-S-glyceryl-cys-ser-lys-lys-lys-lys (Pam 3 Cys-Ser-(Lys) 4 ) [SEQ ID NO: 3]; or   
       MALP-2 dipalmitoyl-S-glyceryl-cys-gly-asn-asn-asp-glu-ser-asn-ile-ser-phe-lys-glu-lys (Pam 2 CGNNDESNISFKEK)] [SEQ ID NO: 4]. 
     
     
         10 - 12 . (canceled) 
     
     
         13 . The composition of  claim 9  comprising mucin 1 (MUC1) variable number tandem repeats (VNTRs) conjugated to tumor-associated carbohydrate antigens (TACA). 
     
     
         14 . The composition of  claim 9 , wherein the antigen is a TACA, selected from the group consisting of TF, Tn, sialyl Tn (sTn), or sialyl Lewis a (sLe a ) antigens, having the formulae: 
       
         
           
           
               
               
           
         
       
     
     
         15 . (canceled) 
     
     
         16 . The composition of  claim 9 , wherein the antigen comprises MUC1 VNTR having one of the following amino acid sequences:
 PDTRPAPGST(Tn)APPAHGVTSA [SEQ ID NO: 1];   TSAPDTRPAPGSTAPPAHGV [SEQ ID NO: 5]; or   TSAPDT(Tn)RPAPGSTAPPAHGV [SEQ ID NO: 6].   
     
     
         17 . (canceled) 
     
     
         18 . The composition of  claim 7 , wherein the first linker (linker a ) comprises a dialkyl-substituted heteroaryl C 1-n  alkyl of Formula VI 
       
         
           
           
               
               
           
         
       
       wherein the “A” group comprises: a chain of C 1-n  alkyl, dialkyl substituted aryl C 1-n  alkyl, or —CH 2 CH 2 (OCH 2 CH 2 ) n —; and n is a positive integer. 
     
     
         19 . (canceled) 
     
     
         20 . A composition of  claim 7 , comprising the compound (9): 
       
         
           
           
               
               
           
         
       
     
     
         21 . A composition of  claim 7 , comprising the compound (17): 
       
         
           
           
               
               
           
         
       
     
     
         22 . A composition of  claim 7 , comprising the compound (21): 
       
         
           
           
               
               
           
         
       
     
     
         23 - 26 . (canceled) 
     
     
         27 . A composition comprising a second lipid (lipid b ) moiety, a second linker (linker b ) moiety, and a xenoantigen moiety, having the Formula VII: 
       
         
           
           
               
               
           
         
       
     
     
         28 . The composition of  claim 27 , wherein the second lipid (lipid b ) moiety contains a structure of the Formula IX: 
       
         
           
           
               
               
           
         
       
     
     
         29 . The composition of  claim 27 , wherein the second linker (linker b ) comprises: a chain of C 1-n  alkyl, dialkyl substituted aryl C 1-n  alkyl, or —CH 2 CH 2 (OCH 2 CH 2 ) n —; wherein n is a positive integer. 
     
     
         30 - 35 . (canceled) 
     
     
         36 . The composition of the  claim 27 , wherein the xenoantigen moiety contains a structure comprising: an α- or β-linked L-rhamnose epitope, a β-linked α-Gal disaccharide epitope, or an α- or β-linked Forssmann disaccharide epitope: 
       
         
           
           
               
               
           
         
       
     
     
         37 . A vaccine composition, comprising:
 1) an antigen composition comprising:
 a first lipid (lipid a ) moiety, a first linker (linker a ) moiety, and an antigen moiety; 
   2) a xenoantigen composition comprising:
 a second lipid (lipid b ) moiety, a second linker (linker b ) moiety, and a xenoantigen moiety; and 
   3) at least one liposomal formulation.   
     
     
         38 - 40 . (canceled) 
     
     
         41 . A vaccine composition of  claim 37 , wherein:
 the antigen composition comprises a Pam 3 Cys-MUC1 VNTR-TACA conjugate;   the second linker (linker b ) moiety comprises a tetraethyleneglycol (TEG) portion; and   the xenoantigen moiety comprises α- or β-linked L-rhamnose.   
     
     
         42 - 59 . (canceled) 
     
     
         60 . A method for improving immunogenicity of vaccines, the method comprising incorporating at least one α- or β-linked L-Rha epitope by direct conjugation or by non-covalent association with at least one liposomal vaccine formulation to increase the immunogenicity of the vaccine by a NA-dependant antigen uptake mechanism.