Antibodies and pharmaceutical compositions containing same useful for inhibiting activity of metalloproteins

Abstract

A method of producing a metalloprotein inhibitor, the method comprising generating antibodies directed at a composition including a metal ion-bound chelator, wherein the composition is selected having structural and electronic properties similar to a functional domain of the metalloprotein, thereby producing the metalloprotein inhibitor.

1 . An antibody comprising an antigen recognition region capable of binding a metal ion bound chelator, said metal ion bound chelator comprising structural and electronic properties similar to a functional domain of a metalloenzyme wherein the antibody inhibits an enzymatic activity of a metalloenzyme and whereas said antigen recognition region does not bind a pro-enzyme form of said metalloenzyme. 
   
   
       2 . The antibody of  claim 1 , wherein said antigen recognition region does not bind said chelator in an absence of said metal ion. 
   
   
       3 . The antibody of  claim 1 , capable of inhibiting 50% of an activity of said metalloenzyme at a concentration less than 10 −7  M. 
   
   
       4 . The antibody of  claim 1 , wherein said metal ion is a transition metal ion selected from the group consisting of Vanadium, Selenium, Molybdenum, Cobalt, Zinc, Copper, Iron, Gallium, Bismuth, Aluminum, Gold, Platinum, Manganese, Chronium, Silver, Antimony, Thallium, Cadmium, Nickel, Mercury and Lead. 
   
   
       5 . The antibody of  claim 1 , wherein said chelator is a polyamine. 
   
   
       6 . The antibody of  claim 5 , wherein said polyamine is at least two histidine molecules. 
   
   
       7 . The antibody of  claim 5 , wherein said polyamine is selected from the group consisting of ethylene diamine, cyclam, porphyrin, diethylenetriamine, triethylenetetramine, triethylenediamine, tetraethylenepentamine, aminoethylethanolamine, aminoethylpiperazine, pentaethylenehexamine, captopril, penicilamine, N,N′-bis(3-aminopropyl)-1,3-propanediamine, N,N′-Bis-(2-aminoethyl)-1,3-propanediamine, 1,7-dioxa-4,10-diazacyclododecane, 1,4,8,11-tetraaza cyclotetradecane-5,7-dione, 1,4,7-triazacyclononane, 1-oxa-4,7,10-triazacyclododecane, 1,4,8,12-tetraazacyclopentadecane, and 1,4,7,10-tetraazacyclododecane. 
   
   
       8 . The antibody of  claim 1 , wherein said metalloenzyme is selected from the group consisting of neutrophil collagenase, collagenase-3, gelatinase A, gelatinase B, stromelysins-2 and 3, matrilysin, macrophage elastase; membrane-type MMPs, aggrecanase, tumor necrosis factor converting enzyme, cytokine convertases, adhesion molecule shedding enzymes, endothelin converting enzyme, angiotensin converting enzyme, neutral endopeptidase, FTSH—bacterial metalloprotease, metallo-lactamase (carbapenases), bacterial toxins and ras farnesyl protein transferase and carbonic anhydrase. 
   
   
       9 . A pharmaceutical composition comprising the antibody of  claim 1  and a physiologically acceptable carrier. 
   
   
       10 . A method of producing a matrix metalloenzyme inhibitor, the method comprising generating antibodies directed at a composition comprising a metal ion-bound chelator, wherein said composition is selected having structural and electronic properties similar to a functional domain of the matrix metalloenzyme, thereby producing the matrix metalloenzyme inhibitor. 
   
   
       11 . The method of  claim 10 , wherein said antibodies are polyclonal antibodies. 
   
   
       12 . The method of  claim 10 , wherein said antibodies are monoclonal antibodies. 
   
   
       13 . The method of  claim 10 , wherein said metal ion is a transition metal ion selected from the group consisting of Vanadium, Selenium, Molybdenum, Cobalt, Zinc, Copper, Iron, Gallium, Bismuth, Aluminum, Gold, Platinum, Manganese, Chronium, Silver, Antimony, Thallium, Cadmium, Nickel, Mercury and Lead. 
   
   
       14 . The method of  claim 10 , wherein said chelator is a polyamine. 
   
   
       15 . The method of  claim 14 , wherein said polyamine is at least two histidine molecules. 
   
   
       16 . The method of  claim 15 , wherein said polyamine is selected from the group consisting of ethylene diamine, cyclam, porphyrin, diethylenetriamine, triethylenetetramine, triethylenediamine, tetraethylenepentamine, aminoethylethanolamine, aminoethylpiperazine, pentaethylenehexamine, captopril, penicilamine, N,N′-bis(3-aminopropyl)-1,3-propanediamine, N,N′-Bis-(2-aminoethyl)-1,3-propanediamine, 1,7-dioxa-4,10-diazacyclododecane, 1,4,8,11-tetraaza cyclotetradecane-5,7-dione, 1,4,7-triazacyclononane, 1-oxa-4,7,10-triazacyclododecane, 1,4,8,12-tetraazacyclopentadecane, and 1,4,7,10-tetraazacyclododecane. 
   
   
       17 . The method of  claim 10 , wherein said composition is selected from the group consisting of tetra-carboxy phenyl porphyrin Co(II), tetra-carboxy phenyl porphyrin Zn(II), metal-bound aliphatic amino group containing compound, metal-bound alicyclic amino group containing compound and metal-bound peptides. 
   
   
       18 . The method of  claim 10 , further comprising testing selectivity of said antibodies towards said chelator, wherein said chelator does not comprise said metal ion. 
   
   
       19 . The method of  claim 10 , further comprising testing selectivity of said antibodies towards a proenzyme form of said matrix metalloprotease. 
   
   
       20 . A method of inhibiting a matrix metalloenzyme activity in a subject in need thereof, the method comprising providing to the subject a therapeutically effective amount of an antibody including an antigen recognition region capable of binding a metal ion bound chelator, wherein said antigen recognition region is not capable of binding a chelator in an absence of said metal ion, said metal ion bound chelator comprising structural and electronic properties similar to a functional domain of a metalloenzyme, thereby inhibiting matrix metalloenzyme activity in the subject. 
   
   
       21 . The method of  claim 20 , wherein said antibody is a polyclonal antibody. 
   
   
       22 . The method of  claim 20 , wherein said antibody is a monoclonal antibody. 
   
   
       23 . The method of  claim 20 , wherein said metal ion is a transition metal ion selected from the group consisting of Vanadium, Selenium, Molybdenum, Cobalt, Zinc, Copper, Iron, Gallium, Bismuth, Aluminum, Gold, Platinum, Manganese, Chronium, Silver, Antimony, Thallium, Cadmium, Nickel, Mercury and Lead. 
   
   
       24 . The method of  claim 20 , wherein said chelator is a polyamine. 
   
   
       25 . The method of  claim 24 , wherein said polyamine is at least two histidine molecules. 
   
   
       26 . The method of  claim 24 , wherein said polyamine is selected from the group consisting of ethylene diamine, cyclam, porphyrin, diethylenetriamine, triethylenetetramine, triethylenediamine, tetraethylenepentamine, aminoethylethanolamine, aminoethylpiperazine, pentaethylenehexamine, captopril, penicilamine, N,N′-bis(3-aminopropyl)-1,3-propanediamine, N,N′-Bis-(2-aminoethyl)-1,3-propanediamine, 1,7-dioxa-4,10-diazacyclododecane, 1,4,8,11-tetraaza cyclotetradecane-5,7-dione, 1,4,7-triazacyclononane, 1-oxa-4,7,10-triazacyclododecane, 1,4,8,12-tetraazacyclopentadecane, and 1,4,7,10-tetraazacyclododecane. 
   
   
       27 . A method of determining if an antibody binds to a metal ion-bound chelator, the method comprising determining conformational changes in binding of the metal ion to the chelator thereof following binding of the antibody, to thereby determine if the antibody binds to the metal ion-bound chelator. 
   
   
       28 . A method of determining if an antibody binds to a metal ion-bound chelator, the method comprising determining electronic changes in the metal ion following binding of the antibody, to thereby determine if the antibody binds to the metal ion-bound chelator.