Multi-component antioxidant compounds, pharmaceutical compositions containing same and their use for reducing or preventing oxidative stress

Abstract

An antioxidant compound is disclosed. The compound is characterized by (a) a peptide including at least three amino acid residues of which at least two are cysteine residues, each having a readily oxidizable sulfhydryl group for effecting antioxidation; and at least two peptide bonds, each being cleavable by at least one intracellular peptidase; and (b) a first hydrophobic or non-charged moiety being attached to an amino terminal of the peptide via a first bond and a second hydrophobic or non-charged moiety being attached to a carboxy terminal of the peptide via a second bond, the first hydrophobic or non-charged moiety and the second hydrophobic or non-charged moiety are selected so as to provide the antioxidant compound with membrane miscibility properties for permitting the antioxidant compound to cross cellular membranes; wherein cleavage of the at least two peptide bonds by the at least one intracellular peptidase results in generation of a plurality of antioxidant species, each including one of the cysteine residues having the readily oxidizable sulfhydryl group and which is also active in effecting antioxidation, thereby providing for a plurality of different antioxidant species acting in synergy in exerting antioxidation.

1 . An antioxidant compound comprising:
 (a) a peptide including at least three amino acid residues of which at least two being cysteine residues each having a readily oxidizable sulfhydryl group for effecting antioxidation; and at least two peptide bonds each being cleavable by at least one intracellular peptidase; and   (b) a first hydrophobic or non-charged moiety being attached to an amino terminal of said peptide via a first bond and a second hydrophobic or non-charged moiety being attached to a carboxy terminal of said peptide via a second bond, said first hydrophobic or non-charged moiety and said second hydrophobic or non-charged moiety are selected so as to provide the antioxidant compound with membrane miscibility properties for permitting the antioxidant compound to cross cellular membranes;   
       wherein cleavage of said at least two peptide bonds by said at least one intracellular peptidase results in generation of several antioxidant species each including at least one of said cysteine residues having said readily oxidizable sulfhydryl group and which is also active in effecting antioxidation, thereby providing for a plurality of different antioxidant species acting in synergy in exerting antioxidation. 
     
     
         2 . The antioxidant compound of  claim 1  having a general formula of: 
       
         
           
                 
                 
               
                     
                   A---Y1---Cys---Y2---Cys---Y3---B 
                 
             
                
               
            
           
         
       
       wherein, Cys is a cysteine residue, A is the first hydrophobic or non-charged moiety; B is the second hydrophobic or non-charged moiety; Y1, Y2 and Y3 are each individually one or more amino acid residues in the range of 0-30 residues, with the provision that Y1, Y2 and Y3 collectively provide for at least two amino acid residues in the peptide. 
     
     
         3 . The antioxidant compound of  claim 2 , wherein A is selected from the group consisting of N-acetyl, tert butyl, iso propyl, n-butyl and n-pentyl. 
     
     
         4 . The antioxidant compound of  claim 2 , wherein B is selected from the group consisting of amide and ester. 
     
     
         5 . The antioxidant compound of  claim 1 , wherein cleavage of said first bond and/or said second bond by a cellular hydrolase results in loosing said membrane miscibility. 
     
     
         6 . The antioxidant compound of  claim 1 , wherein cleavage of said first bond and/or said second bond by a cellular hydrolase results in formation of additional antioxidant species acting in synergy. 
     
     
         7 . The antioxidant compound of  claim 1 , wherein said first bond and said second bond are each independently an ester or peptide bond. 
     
     
         8 . The antioxidant compound of  claim 1 , wherein each of said first hydrophobic or non-charged moiety and said second hydrophobic or non-charged moiety is selected from the group consisting of alkyl, aryl, alkene, arene and cholesteril having a backbone of 2-50 carbon atoms. 
     
     
         9 . The antioxidant compound of  claim 1 , wherein said first hydrophobic or non-charged moiety and said second hydrophobic or non-charged moiety are selected so as to enable the antioxidant compound to cross a blood barrier. 
     
     
         10 . The antioxidant compound of  claim 9 , wherein said blood barrier is selected from the group consisting of a blood brain barrier, a blood retinal barrier and a blood testis barrier. 
     
     
         11 . A pharmaceutical composition for preventing or reducing oxidative stress, the composition comprising a pharmaceutically acceptable carrier and, as an active ingredient, an antioxidant compound, said antioxidant compound including:
 (a) a peptide including at least three amino acid residues of which at least two being cysteine residues, each having a readily oxidizable sulfhydryl group for effecting antioxidation; and at least two peptide bonds each being cleavable by at least one intracellular peptidase; and   (b) a first hydrophobic or non-charged moiety being attached to an amino terminal of said peptide via a first bond and a second hydrophobic or non-charged moiety being attached to a carboxy terminal of said peptide via a second bond, said first hydrophobic or non-charged moiety and said second hydrophobic or non-charged moiety are selected so as to provide the antioxidant compound with membrane miscibility properties for permitting the antioxidant compound to cross cellular membranes;   wherein cleavage of said at least two peptide bonds by said at least one intracellular peptidase results in generation of a plurality of antioxidant species each including at least one of said cysteine residues having said readily oxidizable sulfhydryl group and which is also active in effecting antioxidation, thereby providing for a plurality of different antioxidant species acting in synergy in exerting antioxidation.   
     
     
         12 . The pharmaceutical composition of  claim 11 , wherein said pharmaceutically acceptable carrier is selected from the group consisting of a thickener, a base, a buffer, a diluent, a surface active agent and a preservatives. 
     
     
         13 . The pharmaceutical composition of  claim 11  wherein said antioxidant compound having a general formula of: 
       
         
           
                 
                 
               
                     
                   A---Y1---Cys---Y2---Cys---Y3---B 
                 
             
                
               
            
           
         
       
       wherein, Cys is a cysteine residue, A is the first hydrophobic or non-charged moiety; B is the second hydrophobic or non-charged moiety; Y1, Y2 and Y3 are each individually one or more amino acid residues in the range of 0-30 residues, with the provision that Y1, Y2 and Y3 collectively provide for at least two amino acid residues in the peptide. 
     
     
         14 . The pharmaceutical composition of  claim 13  wherein A is selected from the group consisting of N-acetyl, tert butyl, iso propyl, n-butyl and n-pentyl. 
     
     
         15 . The pharmaceutical composition of  claim 13 , wherein B is selected from the group consisting of amide and ester. 
     
     
         16 . The pharmaceutical composition of  claim 11 , wherein cleavage of said first bond and/or said second bond by a cellular hydrolase results in loosing said membrane miscibility. 
     
     
         17 . The pharmaceutical composition of  claim 11 , wherein cleavage of said first bond and/or said second bond by a cellular hydrolase results in formation of additional antioxidant species acting in synergy. 
     
     
         18 . The pharmaceutical composition of  claim 11 , wherein said first bond and said second bond are each independently an ester or peptide bond. 
     
     
         19 . The pharmaceutical composition of  claim 11 , wherein each of said first hydrophobic or non-charged moiety and said second hydrophobic or non-charged moiety is selected from the group consisting of alkyl, aryl, alkene, arene and cholesteril having a backbone of 14-50 carbon atoms. 
     
     
         20 . The pharmaceutical composition of  claim 11 , wherein said first hydrophobic or non-charged moiety and said second hydrophobic or non-charged moiety are selected so as to enable said antioxidant compound to cross a blood barrier. 
     
     
         21 . The pharmaceutical composition of  claim 20 , wherein said blood barrier is selected from the group consisting of a blood brain barrier, a blood retinal barrier and a blood testis barrier. 
     
     
         22 . A method of treating a disease associated with formation of oxidative stress in a subject, the method comprising locally or systemically administering to the subject an antioxidant compound which comprises:
 (a) a peptide including at least three amino acid residues of which at least two being cysteine residues each having a readily oxidizable sulfhydryl group for effecting antioxidation; and at least two peptide bonds each being cleavable by at least one intracellular peptidase; and   (b) a first hydrophobic or non-charged moiety being attached to an amino terminal of said peptide via a first bond and a second hydrophobic or non-charged moiety being attached to a carboxy terminal of said peptide via a second bond, said first hydrophobic or non-charged moiety and said second hydrophobic or non-charged moiety are selected so as to provide the antioxidant compound with membrane miscibility properties for permitting the antioxidant compound to cross cellular membranes;   
       wherein cleavage of said at least two peptide bonds by said at least one intracellular peptidase results in generation of several antioxidant species each including at least one of said cysteine residues having said readily oxidizable sulfhydryl group and which is also active in effecting antioxidation, thereby providing for a plurality of different antioxidant species acting in synergy in exerting antioxidation. 
     
     
         23 . The method of  claim 22 , wherein said antioxidant compound having a general formula of: 
       
         
           
                 
                 
               
                     
                   A---Y1---Cys---Y2---Cys---Y3---B 
                 
             
                
               
            
           
         
       
       wherein, Cys is a cysteine residue, A is the first hydrophobic or non-charged moiety; B is the second hydrophobic or non-charged moiety; Y1, Y2 and Y3 are each individually one or more amino acid residues in the range of 0-30 residues, with the provision that Y1, Y2 and Y3 collectively provide for at least two amino acid residues in the peptide. 
     
     
         24 . The method of  claim 23 , wherein A is selected from the group consisting of N-acetyl, tert butyl, iso propyl, n-butyl and n-pentyl. 
     
     
         25 . The method of  claim 23 , wherein B is selected from the group consisting of amide and ester. 
     
     
         26 . The method of  claim 22 , wherein cleavage of said first bond and/or said second bond by a cellular hydrolase results in losing said membrane miscibility. 
     
     
         27 . The method of  claim 22 , wherein cleavage of said first bond and/or said second bond by a cellular hydrolase results in formation of additional antioxidant species acting in synergy. 
     
     
         28 . The method of  claim 22 , wherein said first bond and said second bond are each independently an ester or peptide bond. 
     
     
         29 . The method of  claim 22 , wherein each of said first hydrophobic or non-charged moiety and said second hydrophobic or non-charged moiety is selected from the group consisting of alkyl, aryl, alkene, arene and cholesteril having a backbone of 2-50 carbon atoms. 
     
     
         30 . The method of  claim 22 , wherein said first hydrophobic or non-charged moiety and said second hydrophobic or non-charged moiety are selected so as to enable the method to cross a blood barrier. 
     
     
         31 . The method of  claim 30 , wherein said blood barrier is selected from the group consisting of a blood brain barrier, a blood retinal barrier and a blood testis barrier. 
     
     
         32 . The method of  claim 22 , wherein the disease associated with formation of oxidative stress is a central nervous system disease. 
     
     
         33 . The method of  claim 32 , wherein said central nervous system disease is selected from the group comprising a neurodegenerative disorder, Parkinson's disease, Alzheimer's disease, Creutzfeldt-Jakob disease, cerebral ischemia, multiple sclerosis, a degenerative disease of the basal ganglia, a motoneuron disease, scrapies, spongiform encephalopathy, a neurological viral disease, a motoneuron disease, post-surgical neurological dysfunction, memory loss and memory impairment. 
     
     
         34 . The method of  claim 22 , wherein the disease associated with formation of oxidative stress is a non-central nervous system disease. 
     
     
         35 . The method of  claim 34 , wherein said non-central nervous system disease is selected from the group comprising rheumatoid arthritis, cataract, Down syndrome, cystic fibrosis, diabetes, acute respiratory distress syndrome, asthma, post-surgical neurological dysfunction, amyotrophic lateral sclerosis, atherosclerotic cardiovascular disease, hypertension, post-operative restenosis, pathogenic vascular smooth muscle cell proliferation, pathogenic intra-vascular macrophage adhesion, pathogenic platelet activation, pathogenic lipid peroxidation, myocarditis, stroke, multiple organ dysfunction, complication resulting from inflammatory processes, AIDS, cancer, aging, bacterial infection, sepsis; viral disease, AIDS, hepatitis C, influenza and a neurological viral disease. 
     
     
         36 . A method of treating a habit associated with formation of oxidative stress in a subject, the method comprising locally or systemically administering to the subject an antioxidant compound which comprises:
 (a) a peptide including at least three amino acid residues of which at least two being cysteine residues each having a readily oxidizable sulfhydryl group for effecting antioxidation; and at least two peptide bonds each being cleavable by at least one intracellular peptidase; and   (b) a first hydrophobic or non-charged moiety being attached to an amino terminal of said peptide via a first bond and a second hydrophobic or non-charged moiety being attached to a carboxy terminal of said peptide via a second bond, said first hydrophobic or non-charged moiety and said second hydrophobic or non-charged moiety are selected so as to provide the antioxidant compound with membrane miscibility properties for permitting the antioxidant compound to cross cellular membranes;   
       wherein cleavage of said at least two peptide bonds by said at least one intracellular peptidase results in generation of several antioxidant species each including at least one of said cysteine residues having said readily oxidizable sulfhydryl group and which is also active in effecting antioxidation, thereby providing for a plurality of different antioxidant species acting in synergy in exerting antioxidation. 
     
     
         37 . The method of  claim 22 , wherein said antioxidant compound having a general formula of: 
       
         
           
                 
                 
               
                     
                   A---Y1---Cys---Y2---Cys---Y3---B 
                 
             
                
               
            
           
         
       
       wherein, Cys is a cysteine residue, A is the first hydrophobic or non-charged moiety; B is the second hydrophobic or non-charged moiety; Y1, Y2 and Y3 are each individually one or more amino acid residues in the range of 0-30 residues, with the provision that Y1, Y2 and Y3 collectively provide for at least two amino acid residues in the peptide. 
     
     
         38 . The method of  claim 23 , wherein A is selected from the group consisting of N-acetyl, tert butyl, iso propyl, n-butyl and n-pentyl. 
     
     
         39 . The method of  claim 23 , wherein B is selected from the group consisting of amide and ester. 
     
     
         40 . The method of  claim 22 , wherein cleavage of said first bond and/or said second bond by a cellular hydrolase results in loosing said membrane miscibility. 
     
     
         41 . The method of  claim 22 , wherein cleavage of said first bond and/or said second bond by a cellular hydrolase results in formation of additional antioxidant species acting in synergy. 
     
     
         42 . The method of  claim 22 , wherein said first bond and said second bond are each independently an ester or peptide bond. 
     
     
         43 . The method of  claim 22 , wherein each of said first hydrophobic or non-charged moiety and said second hydrophobic or non-charged moiety is selected from the group consisting of alkyl, aryl, alkene, arene and cholesteril having a backbone of 2-50 carbon atoms. 
     
     
         44 . The method of  claim 22 , wherein said first hydrophobic or non-charged moiety and said second hydrophobic or non-charged moiety are selected so as to enable the method to cross a blood barrier. 
     
     
         45 . The method of  claim 30 , wherein said blood barrier is selected from the group consisting of a blood brain barrier, a blood retinal barrier and a blood testis barrier. 
     
     
         46 . The method of  claim 22 , wherein the habit associated with formation of oxidative stress is selected from the group comprising aging, smoking, sun tanning, cancer treatment, radiation, cocaine consumption and morphine consumption. 
     
     
         47 . The method of  claim 22 , wherein said antioxidant compound is administered in a pharmaceutical composition which includes a pharmaceutically acceptable carrier. 
     
     
         48 . The method of  claim 47 , wherein said pharmaceutically acceptable carrier adapts the composition for administration by a route selected from the intranasal, transdermal, intradermal, oral, buccal, parenteral, topical, rectal and inhalation route. 
     
     
         49 . The method of  claim 47 , wherein the carrier provides said antioxidant compound in solution, suspension, emulsion, gel or skin pad. 
     
     
         50 . The method of  claim 47 , wherein the composition further includes a formulating agent selected from the group consisting of a suspending agent, a stabilizing agent and a dispersing agent.