Angiotensin peptide-carrier conjugates and uses thereof

Abstract

The present invention provides conjugates of peptide derivatives of the mammalian peptide hormones angiotensinogen, angiotensin I and angiotensin II, presented in a repetitive scaffold by coupling the peptide derivatives to a carrier, particularly a virus-like particle (VLP). The invention also provides methods of producing such conjugates, and immunotherapeutic uses of the resulting immunogen conjugates for the therapy and prophylaxis of conditions associated with the renin-activated angiotensin system.

1 . (canceled) 
     
     
         2 . (canceled) 
     
     
         3 . An angiotensin peptide moiety-carrier conjugate comprising:
 (a) a carrier with at least one first attachment site, and   (b) at least one angiotensin peptide moiety with at least one second attachment site;   wherein said carrier comprises a core particle that is a virus-like particle and;   wherein said second attachment site is capable of association through at least one covalent bond to said first attachment site so as to form an ordered and repetitive angiotensin peptide moiety-carrier conjugate.   
     
     
         4 . The conjugate of  claim 3 , wherein said virus-like particle comprises recombinant proteins or fragments thereof selected from the group consisting of:
 (a) recombinant proteins of Hepatitis B virus;   (b) recombinant proteins of measles virus;   (c) recombinant proteins of Sindbis virus;   (d) recombinant proteins of Rotavirus;   (e) recombinant proteins of Foot-and-Mouth-Disease virus;   (f) recombinant proteins of Retrovirus;   (g) recombinant proteins of Norwalk virus;   (h) recombinant proteins of Alphavirus;   (i) recombinant proteins of human Papilloma virus;   (j) recombinant proteins of Polyoma virus;   (k) recombinant proteins of bacteriophages;   (l) recombinant proteins of RNA-phages;   (m) recombinant proteins of Qβ-phage;   (n) recombinant proteins of GA-phage   (o) recombinant proteins of fr-phage   (p) recombinant proteins of AP205 phage; and   (q) recombinant proteins of Ty.   
     
     
         5 . (canceled) 
     
     
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         7 . (canceled) 
     
     
         8 . The conjugate of  claim 3 , wherein said virus or virus-like particle comprises proteins or fragments thereof of an RNA-bacteriophage. 
     
     
         9 . The conjugate of  claim 8 , wherein said RNA-bacteriophage is selected from the group consisting of:
 a) bacteriophage Qβ;   b) bacteriophage R17;   c) bacteriophage fr;   d) bacteriophage GA;   e) bacteriophage SP;   f) bacteriophage MS2;   g) bacteriophage M11;   (h) bacteriophage MX1;   (i) bacteriophage NL95;   (j) bacteriophage f2;   (k) bacteriophage AP205; and   (l) bacteriophage PP7.   
     
     
         10 . The conjugate of  claim 3 , wherein said virus-like particle comprises recombinant proteins or fragments thereof of bacteriophage Qβ. 
     
     
         11 . (canceled) 
     
     
         12 . The conjugate of  claim 8 , wherein said recombinant proteins of said RNA-bacteriophage comprise mutant coat proteins. 
     
     
         13 . The conjugate of  claim 12 , wherein said mutant coat proteins have been modified (i) by removal of at least one lysine residue by way of substitution, or (ii) by addition of at least one lysine residue by way of substitution, or (iii) by deletion of at least one lysine residue, or (iv) by addition of at least one lysine residue by way of insertion. 
     
     
         14 . (canceled) 
     
     
         15 . The conjugate of  claim 10 , wherein said recombinant proteins comprise one or more coat proteins having an amino acid sequence of SEQ ID NO:3, or comprise a mixture of coat proteins having amino acid sequences of SEQ ID NO: 4 or mutants thereof, and SEQ ID NO:3. 
     
     
         16 . (canceled) 
     
     
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         21 . The conjugate of  claim 3 , wherein said virus-like particle comprises one or more mutant Qβ coat proteins having an amino acid sequence selected from the group consisting of:
 a) the amino acid sequence of SEQ ID NO:6; 
 b) the amino acid sequence of SEQ ID NO:7; 
 c) the amino acid sequence of SEQ ID NO:8; 
 d) the amino acid sequence of SEQ ID NO:9; and 
 e) the amino acid sequence of SEQ ID NO:10. 
 
     
     
         22 . The conjugate of  claim 3 , wherein said first attachment site comprises:
 a) an amino group;   b) a carboxyl group;   c) a sulfhydryl group;   d) a hydroxy group;   e) a guanidinyl group; or   f) a histidinyl group.   
     
     
         23 . The conjugate of  claim 3 , wherein said at least one first attachment site is selected from the group consisting of a lysine residue, an arginine residue, a cysteine residue, an aspartate, a glutamate residue, a serine residue, a threonine residue, a histidine residue and a tyrosine residue. 
     
     
         24 . (canceled) 
     
     
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         26 . (canceled) 
     
     
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         30 . (canceled) 
     
     
         31 . (canceled) 
     
     
         32 . (canceled) 
     
     
         33 . (canceled) 
     
     
         34 . The conjugate of  claim 3 , wherein said angiotensin peptide moiety is an angiotensin peptide selected from the group consisting of angiotensinogen, angiotensin I, angiotensin II, and fragments or derivatives thereof. 
     
     
         35 . The conjugate of  claim 3 , wherein said angiotensin peptide moiety with said second attachment site has an amino acid sequence selected from the group consisting of: 
       
         
           
                 
                 
                 
               
                     
                   a) 
                   CGGDRVYIHPF; 
                 
                     
                     
                 
                     
                   b) 
                   CGGDRVYIHPFHL; 
                 
                     
                     
                 
                     
                   c) 
                   DRVYIHPFHLGGC; 
                 
                     
                     
                 
                     
                   d) 
                   CDRVYIHPFHL; 
                 
                     
                     
                 
                     
                   e) 
                   CHPFHL; 
                 
                     
                     
                 
                     
                   f) 
                   CGPFHL; 
                 
                     
                     
                 
                     
                   g) 
                   CYIHPF; 
                 
                     
                     
                 
                     
                   h) 
                   CGIHPF; 
                 
                     
                     
                 
                     
                   i) 
                   CGGHPF; 
                 
                     
                     
                 
                     
                   j) 
                     D RVYIGGC; 
                 
                     
                     
                 
                     
                   k) 
                   DRVYGGC; 
                 
                     
                   and 
                     
                 
                     
                     
                 
                     
                   l) 
                     DRV GGC. 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         36 . A pharmaceutical composition comprising one or more of the conjugates of  claim 3  and a pharmaceutically acceptable carrier or excipient. 
     
     
         37 . A vaccine composition comprising an immunologically effective amount of the conjugate of  claim 3  and an immunologically acceptable carrier or excipient. 
     
     
         38 . The vaccine composition of  claim 37 , wherein said vaccine composition further comprises at least one adjuvant. 
     
     
         39 . A method of immunizing an animal against an angiotensin peptide moiety, comprising administering the conjugate of  claim 3  to an animal under conditions such that said animal develops an immune response to said angiotensin peptide moiety. 
     
     
         40 . The method of  claim 39 , wherein said conjugate or said composition is administered to said animal via a route of administration selected from the group consisting of intranasal administration, oral administration, subcutaneous administration, transdermal administration, intramuscular administration and intravenous administration. 
     
     
         41 . (canceled) 
     
     
         42 . (canceled) 
     
     
         43 . (canceled) 
     
     
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         45 . A method of treating or preventing a physical disorder associated with the renin-activated angiotensin system comprising administering to an animal in need thereof a therapeutically or prophylactically effective amount of one or more of the conjugates of  claim 3 . 
     
     
         46 . The method of  claim 45 , wherein said physical disorder associated with the renin-activated angiotensin system is selected from the group consisting of hypertension, stroke, infarction, congestive heart failure, kidney failure and retinal hemorrhage. 
     
     
         47 . (canceled) 
     
     
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         49 . (canceled)