Nitrosylation of protein SH groups and amino acid residues as a therapeutic modality

Abstract

Nitrosylation of proteins and amino acid groups enables selective regulation of protein function, and also endows the proteins and amino acids with additional smooth muscle relaxant and platelet inhibitory capabilities. Thus, the invention relates to novel compounds achieved by nitrosylation of protein thiols. Such compounds include: S-nitroso-t-PA, S-nitroso-cathepsin; S-nitroso-lipoprotein; and S-nitroso-immunoglobulin. The invention also relates to therapeutic use of S-nitroso-protein compounds for regulating protein function, cellular metabolism and effecting vasodilation, platelet inhibition, relaxation of non-vascular smooth muscle, and increasing blood oxygen transport by hemoglobin and myoglobin. The compounds are also used to deliver nitric oxide in its most bioactive form in order to achieve the effects described above, or for in vitro nitrosylation of molecules present in the body. The invention also relates to the nitrosylation of oxygen, carbon and nitrogen moieties present on proteins and amino acids, and the use thereof to achieve the above physiological effects.

1 . A method for causing vasodilation or relaxing non-vascular smooth muscle comprising administering to an animal in need thereof a therapeutically effective amount of a pharmaceutical composition comprising an S-nitroso-enzyme, an S-nitroso-lipoprotein, an S-nitroso-immunoglobulin, an S-nitroso-hemoglobin or an S-nitroso-albumin.  
   
   
       2 . The method of  claim 2 , wherein the smooth muscle comprises airway smooth muscle.  
   
   
       3 . A method for treating a cardiovascular disorder or a respiratory disorder comprising administering to an animal in need thereof a therapeutically effective amount of a pharmaceutical composition comprising an S-nitroso-enzyme, an S-nitroso-lipoprotein, an S-nitroso-immunoglobulin, an S-nitroso-hemoglobin or an S-nitroso-albumin.  
   
   
       4 . A method for lysing a blood clot comprising administrating to an animal in need thereof a therapeutically effective amount of a pharmaceutical composition comprising an S-nitroso-tissue-type plasminogen activator, an S-nitroso-streptokinase, or an S-nitroso-urokinase.  
   
   
       5 . A method for regulating protein or amino acid function or for preventing cellular uptake of one or more proteins comprising administering a therapeutic amount of a nitrosylating compound to an animal in need thereof.  
   
   
       6 . The method of  claim 5 , wherein the nitrosylating compound is nitroglycerin, a nitrosothiol, or nitric oxide.  
   
   
       7 . A method for regulating the function of proteins in which a thiol is bound to a methyl group comprising: 
 removing the methyl groups from the thiol by selective de-methylation; and    reacting the free thiol group with a nitrosylating agent.    
   
   
       8 . A method for regulating the function of a protein which lacks a free thiol group comprising: 
 adding a thiol group to the protein by chemical or genetic engineering means; and    reacting the thiol group with a nitrosylating agent.    
   
   
       9 . A method for regulating cellular function comprising S-nitrosylating a protein, wherein the protein a cellular component or affects cellular function.  
   
   
       10 . The method of  claim 9 , wherein the protein is a cell receptor, G-protein, a target protein, histone, a protein involved in cell proliferation, a protein involved in inhibition of proliferation, a protein involved in cellular repair, an immune modulator, a protein with a cytostatic function or a protein with a cytotoxic function.  
   
   
       11 . A method for delivering nitric oxide to one or more specific targeted sites in a body comprising administering to an animal in need thereof a therapeutically effective amount of a pharmaceutical composition comprising an S-nitroso-enzyme, an S-nitroso-lipoprotein, an S-nitroso-immunoglobulin, an S-nitroso-hemoglobin or an S-nitroso-albumin.  
   
   
       12 . A method for inhibiting platelet function comprising administering to an animal in need thereof a therapeutically effective amount of a pharmaceutical composition comprising an S-nitroso-lipoprotein, an S-nitroso-immunoglobulin, or an S-nitroso-hemoglobin.  
   
   
       13 . The method of  claim 12 , wherein the pharmaceutical composition further comprises a pharmaceutically acceptable carrier.  
   
   
       14 . The method of  claim 12 , wherein the S-nitroso-lipoprotein has at least one nitrogen monoxide moiety bonded at a sulfur atom, an oxygen atom or a nitrogen atom on the lipoprotein.  
   
   
       15 . The method of  claim 14 , wherein the lipoprotein is a chylomicron, a chylomicron remnant particle, a very low-density lipoprotein, an intermediate-density lipoprotein, a low-density lipoprotein, a high density lipoprotein or a lipoprotein (a).  
   
   
       16 . The method of  claim 12 , wherein the S-nitroso-hemoglobin has at least one nitrogen monoxide moiety bonded at a sulfur atom, an oxygen atom or a nitrogen atom on the hemoglobin.  
   
   
       17 . The method of  claim 16 , wherein the hemoglobin is myoglobin.  
   
   
       18 . The method of  claim 12 , wherein the S-nitroso-immunoglobulin has at least one nitrogen monoxide moiety bonded at a sulfur atom, an oxygen atom or a nitrogen atom on the immunoglobulin.  
   
   
       19 . The method of  claim 18 , wherein the immunoglobulin molecule is IgG, IgM, IgA, IgD or IgE.