Caveolin peptides and their use as therapeutics

Abstract

The present invention relates generally to compositions and methods useful for treating various conditions and afflictions, such as inflammation and cancer. More specifically, the present invention relates to compositions and methods of treatment which utilize peptides comprising at least one caveolin scaffolding domain. Even more specifically, the present invention relates to compositions of fusion peptides comprising the antennapedia homeodomain fused to a caveolin scaffolding domain and to methods of using these peptides to treat various conditions and afflictions.

1 . A method of blocking the interaction of a caveolin with a protein in vivo wherein the method comprises administering an effective amount of a peptide which comprises a caveolin scaffolding domain.  
     
     
         2 . The method of  claim 1  wherein the interaction comprises the binding of a protein to a caveolin.  
     
     
         3 . A method of down regulating a caveolin-binding protein in vivo wherein the method comprises administering an effective amount of a peptide which comprises a caveolin scaffolding domain.  
     
     
         4 . A method of inhibiting inflammation in an animal wherein the method comprises administering an effective amount of a peptide which comprises a caveolin scaffolding domain.  
     
     
         5 . A method of inhibiting tumor cell angiogenesis proliferation in an animal wherein the method comprises administering an effective amount of a peptide which comprises a caveolin scaffolding domain.  
     
     
         6 . The method of  claim 5  wherein the caveolin scaffolding domain is selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID No: 4.  
     
     
         7 . The method of  claim 1 ,  2 ,  3 ,  4 ,  5  or  6  wherein the peptide further comprises a membrane translocation domain.  
     
     
         8 . The method of  claim 7  wherein the membrane translocation domain comprises the third helix of the antennapedia homeodomain.  
     
     
         9 . The method of  claim 7  wherein the membrane translocation domain comprises the amino acid sequence in SEQ ID NO: 10.  
     
     
         10 . The method of  claim 1 ,  2 ,  3 ,  4 ,  5  or  6  wherein the protein is eNOS.  
     
     
         11 . The method of  claim 1 ,  2 ,  3 ,  4  or  5  wherein the peptide comprises the amino acid sequence of SEQ ID NO: 11.  
     
     
         12 . A fusion peptide comprising at least one caveolin scaffolding domain and at least one membrane translocation domain.  
     
     
         13 . The fusion peptide of  claim 12  wherein the membrane translocation domain facilitates membrane translocation in vivo.  
     
     
         14 . The fusion peptide of  claim 12  wherein the membrane translocation domain comprises the third helix of the antennapedia homeodomain.  
     
     
         15 . The fusion peptide of  claim 12  wherein the caveolin scaffolding domain is selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4.  
     
     
         16 . An isolated peptide selected from the group consisting of: 
 (a) an isolated peptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 11 and SEQ ID NO: 12    (b) an isolated peptide comprising a fragment of at least three amino acids of an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 11 and SEQ ID NO: 12;    (c) an isolated peptide comprising conservative amino acid substitutions of the amino acid sequences selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 11 and SEQ ID NO: 12; and    (d) naturally occurring amino acid sequence variants of amino acid sequences selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 11 and SEQ ID NO: 12.    
     
     
         17 . A composition comprising the fusion peptide of  claim 12  or the peptide of  claim 16 .  
     
     
         18 . The composition of  claim 17  further comprising a carrier.  
     
     
         19 . An isolated peptide consisting of the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 11 or SEQ ID NO: 12.  
     
     
         20 . An isolated peptide consisting of the amino acid sequence of SEQ ID NO: 11.  
     
     
         21 . A method of down-regulating eNOS activity in a cell comprising administering an effective amount of a peptide which down-regulates at least one activity of eNOS.  
     
     
         22 . The method of  claim 21  wherein the peptide comprises at least one caveolin scaffolding domain.  
     
     
         23 . The method of  claim 22  wherein the peptide further comprises at least one membrane translocation domain.  
     
     
         24 . The method of  claim 21  wherein the peptide is selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4.  
     
     
         25 . The method of  claim 21  wherein the eNOS activity is synthesis of NO.  
     
     
         26 . The method of  claim 21  wherein the down-regulation of eNOS activity results in blockade of vasodilation.  
     
     
         27 . A method of inhibiting inflammation in a mammal comprising administering an effective amount of a peptide which down-regulates at least one activity of eNOS.  
     
     
         28 . A method of inhibiting tumor cell angiogenesis and proliferation in a mammal comprising administering an effective amount of a peptide which down-regulates at least one activity of eNOS.  
     
     
         29 . The method of  claim 28  wherein the peptide is administered in combination with a chemotherapeutic agent.  
     
     
         30 . A method of inhibiting eNOS-dependent vasodilation in a mammal comprising administering an effective amount of a peptide which down-regulates at least one activity of eNOS.  
     
     
         31 . The method of  claim 27 ,  28 ,  29  or  30  wherein the peptide is a fusion peptide comprising at least one caveolin scaffolding domain.  
     
     
         32 . The method of  claim 31  wherein the fusion peptide further comprises at least one membrane translocation domain.  
     
     
         33 . The method of  claim 31  wherein the caveolin scaffolding domain is selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO:4.  
     
     
         34 . The method of  claim 27 ,  28 ,  29  or  30  wherein the eNOS activity is NO production.  
     
     
         35 . A method of identifying an agent which interacts with eNOS comprising: 
 (a) exposing cells which express eNOS to an agent; and    (b) determining if the agent binds to eNOS, thereby identifying an agent which interacts with eNOS.    
     
     
         36 . A method of identifying an agent which modulates at least one activity of eNOS comprising the steps of: 
 (a) exposing cells which express eNOS to an agent; and    (b) determining whether the agent modulates the activity of eNOS, thereby identifying an agent which modulates an activity of eNOS.    
     
     
         37 . The method of  claim 36  wherein the activity is selected from the group consisting of acetycholine-induced vasodilation, prostacyclin production and NO production.