Novel polyamine analog conjugates and quinone conjugates as therapies for cancers and prostate diseases

Abstract

Peptide conjugates in which cytocidal and cytostatic agents, such as polyamine analogs or naphthoquinones, are conjugated to a polypeptide recognized and cleaved by enzymes such as prostate-specific antigen (PSA) and cathepsin B are provided, as well as compositions comprising these conjugates. Methods of using these conjugates in the treatment of prostate diseases are also provided.

What is claimed is:  
     
         1 . A therapeutic composition comprising a peptide substrate covalently attached to a cytostatic or cytocidal agent wherein an enzyme catalyzes cleavage of the peptide substrate from the cytostatic or cytocidal agent.  
     
     
         2 . A composition according to  claim 1  wherein the cytostatic or cytocidal agent is a polyamine.  
     
     
         3  A composition according to  claim 2 , wherein the polyamine analog is linked to the peptide at the carboxy terminus of the peptide by an amide linkage to a primary or secondary amine of the polyamine.  
     
     
         4  A composition according to  claim 2 , wherein the polyamine analog contains a hydroxy group and is linked to the peptide at the carboxy terminus of the peptide by an ester linkage through said hydroxy group.  
     
     
         5  A composition according to  claim 2 , wherein the polyamine analog is of the formula:  
       —N(—E)—B—A—B—NH—B—A—B—NH—B—A—B—NH—B—A—B—NH—E or —HN—B—A—B—NH—B—A—B—NH—B—A—B—NH—B—A—B—NH—E  wherein A is independently selected from the group consisting of: a single bond, C 2 -C 6  alkenyl, C 2 -C 6  alkynyl, C 3 -C 6  cycloalkyl, C 3 -C 6  cycloaryl, and C 3 -C 6  cycloalkenyl;    B is independently selected from the group consisting of: a single bond, C 1 -C 6  alkyl, and C 2 -C 6  alkenyl;    and E is independently selected from the group consisting of H, C 1 -C 6  alkyl, C 2 -C 6  alkenyl, C 2 -C 6  alkynyl, C 3 -C 6  cycloalkyl, C 3 -C 6  cycloaryl, and C 3 -C 6  cycloalkenyl;    and any salt or stereoisomer thereof.    
     
     
         6  A composition according to  claim 5 , wherein the polyamine analog is of the formula:  
       —N(—E)—B—A—B—NH—B—A—B—NH—B—A—B—NH—B—A—B—NH—E or —HN—B—A—B—NH—B—A—B—NH—B—A—B—NH—B—A—B—NH—E  wherein A is independently selected from the group consisting of: a single bond, C 2 -C 6  alkenyl, C 2 -C 6  alkynyl, C 3 -C 6  cycloalkyl, C 3 -C 6  cycloaryl, and C 3 -C 6  cycloalkenyl;    B is independently selected from the group consisting of: a single bond, C 1 -C 6  alkyl, and C 2 -C 6  alkenyl;    and E is independently selected from the group consisting of H, C 1 -C 6  alkyl, C 2 -C 6  alkenyl, C 2 -C 6  alkynyl, C 3 -C 6  cycloalkyl, C 3 -C 6  cycloaryl, and C 3 -C 6  cycloalkenyl;    with the proviso that either at least one A moiety is selected from the group consisting of C 2 -C 6  alkenyl, C 2 -C 6  alkynyl, C 3 -C 6  cycloalkyl, C 3 -C 6  cycloaryl, and C 3 -C 6  cycloalkenyl, or at least one B moiety is selected from the group consisting of C 2 -C 6  alkenyl;    and any salt or stereoisomer thereof.    
     
     
         7  A composition according to  claim 2  wherein the polyamine analog is of the formula:  
       —N(—E)—B—A—B—NH—B—A—B—NH—B—A—B—NH(—B—A—B—NH) x —E or —HN—B—A—B—NH—B—A—B—NH—B—A—B—NH(—B—A—B—NH) x —E  wherein A is independently selected from the group consisting of: a single bond, C 6 -C 2  alkyl, C 2 -C 6  alkenyl, C 2 -C 6  alkynyl, C 3 -C 6  cycloalkyl, C 3 -C 6  cycloaryl, and C 3 -C 6  cycloalkenyl;    B is independently selected from the group consisting of: a single bond, C 1 -C 6  alkyl, and C 2 -C 6  alkenyl;    E is independently selected from the group consisting of H, C 1 -C 6  alkyl, C 2 -C 6  alkenyl, C 2 -C 6  alkynyl, C 3 -C 6  cycloalkyl, C 3 -C 6  cycloaryl, and C 3 -C 6  cycloalkenyl; and x is an integer from 2 to 16;    and any salt or stereoisomer thereof.    
     
     
         8 . A composition according to  claim 7  wherein the polyamine analog is of the formula:  
       —N(—E)—B—A—B—NH—B—A—B—NH—B—A—B—NH(—B—A—B—NH) x —E or —HN—B—A—B—NH—B—A—B—NH—B—A—B—NH(—B—A—B—NH) x —E  wherein A is independently selected from the group consisting of: a single bond, C 6 -C 2  alkyl, C 2 -C 6  alkenyl, C 2 -C 6  alkynyl, C 3 -C 6  cycloalkyl, C 3 -C 6  cycloaryl, and C 3 -C 6  cycloalkenyl;    B is independently selected from the group consisting of: a single bond, C 1 -C 6  alkyl, and C 2 -C 6  alkenyl;    E is independently selected from the group consisting of H, C 1 -C 6  alkyl, C 2 -C 6  alkenyl, C 2 -C 6  alkynyl, C 3 -C 6  cycloalkyl, C 3 -C 6  cycloaryl, and C 3 -C 6  cycloalkenyl; and x is an integer from 2 to 16;    with the proviso that either at least one A moiety is selected from the group consisting of C 2 -C 6  alkenyl, C 2 -C 6  alkynyl, C 3 -C 6  cycloalkyl, C 3 -C 6  cycloaryl, and C 3 -C 6  cycloalkenyl, or at least one B moiety is selected from the group consisting of C 2 -C 6  alkenyl;    and any salt or stereoisomer thereof.    
     
     
         9 . A composition according to  claim 2  wherein the polyamine analog is of the formula:  
       E—NH—B—A—B—NH—B—A—B—NH—B—A—B—NH(—B—A—B—NH) x —E  wherein A is independently selected from the group consisting of: a single bond, C 2 -C 6  alkenyl, C 2 -C 6  alkynyl, C 3 -C 6  cycloalkyl, C 3 -C 6  cycloaryl, and C 3 -C 6  cycloalkenyl;    B is independently selected from the group consisting of: a single bond, C 1 -C 6  alkyl, and C 2 -C 6  alkenyl;    E is independently selected from the group consisting of C 1 -C 6  alkyl, C 1 -C 6  alkanol, C 3 -C 6  cycloalkanol, and C 3 -C 6  hydroxyaryl, and the peptide is linked to the polyamine via an ester linkage at one and only one E group hydroxy;    and x is an integer from 0 to 16;    and any salt or stereoisomer thereof.    
     
     
         10 . A composition according to  claim 9  wherein the polyamine analog is of the formula:  
       E—NH—B—A—B—NH—B—A—B—NH—B—A—B—NH(—B—A—B—NH) x —E  wherein A is independently selected from the group consisting of: a single bond, C 2 -C 6  alkenyl, C 2 -C 6  alkynyl, C 3 -C 6  cycloalkyl, C 3 -C 6  cycloaryl, and C 3 -C 6  cyCloalkenyl;    B is independently selected from the group consisting of: a single bond, C 1 -C 6  alkyl, and C 2 -C 6  alkenyl;    E is independently selected from the group consisting of C 1 -C 6  alkyl, C 1 -C 6  alkanol, C 3 -C 6  cycloalkanol, and C 3 -C 6  hydroxyaryl, with the proviso that at least one E moiety be selected from the group consisting of C 1 -C 6  alkanol, C 3 -C 6  cycloalkanol, and C 3 -C 6  hydroxyaryl, and the peptide is linked to the polyamine via an ester linkage at one and only one E group hydroxy;    and x is an integer from 0 to 16;    and any salt or stereoisomer thereof.    
     
     
         11 . A composition according to  claim 2  wherein the polyamine analog is of the formula:  
       —N(—E)—D—NH—B—A—B—NH—D—NH—E or —NH—D—NH—B—A—B—NH—D—NH—E  wherein A is selected from the group consisting of C 2 -C 6  alkynyl;    B is independently selected from the group consisting of: a single bond, C 1 -C 6  alkyl, and C 2 -C 6  alkenyl;    D is independently selected from the group consisting of C 1 -C 6  alkyl, C 2 -C 6  alkenyl, C 2 -C 6  alkynyl, C 3 -C 6  cycloalkyl, C 3 -C 6  cycloalkenyl, and C 3 -C 6  cycloaryl; and    E is independently selected from the group consisting of H, C 1 -C 6  alkyl, C 2 -C 6  alkenyl, C 2 -C 6  alkynyl, C 3 -C 6  cycloalkyl, C 3 -C 6  cycloaryl, and C 3 -C 6  cycloalkenyl;    and any salt or stereoisomer thereof.    
     
     
         12 . A composition according to  claim 2  wherein the polyamine analog is of the formula:  
       —N(—E)—B—A—B—NH—F—NH—B—A—B—NH—E or —NH—B—A—B—NH—F—NH—B—A—B—NH—E  wherein F is selected from the group consisting of C 1 -C 6  alkyl;    A is independently selected from the group consisting of: a single bond, C 1 -C 6  alkyl; C 2 -C 6  alkenyl, C 2 -C 6  alkynyl, C 3 -C 6  cycloalkyl, C 3 -C 6  cycloaryl, and C 3 -C 6  cycloalkenyl;    B is independently selected from the group consisting of: a single bond, C 1 -C 6  alkyl, and C 2 -C 6  alkenyl; and    E is independently selected from the group consisting of H, C 1 -C 6  alkyl, C 2 -C 6  alkenyl, C 2 -C 6  alkynyl, C 3 -C 6  cycloalkyl, C 3 -C 6  cycloaryl, and C 3 -C 6  cycloalkenyl;    and any salt or stereoisomer thereof.    
     
     
         13 . A composition according to  claim 12  wherein the polyamine analog is of the formula:  
       —N(—E)—B—A—B—NH—F—NH—B—A—B—NH—E or —NH—B—A—B—NH—F—NH—B—A—B—NH—E  wherein F is selected from the group consisting of C 1 -C 6  alkyl;    A is independently selected from the group consisting of: a single bond, C 1 -C 6  alkyl; C 2 -C 6  alkenyl, C 2 -C 6  alkynyl, C 3 -C 6  cycloalkyl, C 3 -C 6  cycloaryl, and C 3 -C 6  cycloalkenyl;    B is independently selected from the group consisting of: a single bond, C 1 -C 6  alkyl, and C 2 -C 6  alkenyl; and    E is independently selected from the group consisting of H, C 1 -C 6  alkyl, C 2 -C 6  alkenyl, C 2 -C 6  alkynyl, C 3 -C 6  cycloalkyl, C 3 -C 6  cycloaryl, and C 3 -C 6  cycloalkenyl;    with the proviso that either at least one A moiety is selected from the group consisting of C 2 -C 6  alkenyl, C 2 -C 6  alkynyl, C 3 -C 6  cycloalkyl, C 3 -C 6  cycloaryl, and C 3 -C 6  cycloalkenyl, or at least one B moiety is selected from the group consisting of C 2 -C 6  alkenyl;    and any salt or stereoisomer thereof.    
     
     
         14 . A composition according to  claim 1  wherein the cytostatic or cytocidal agent is a quinone.  
     
     
         15 . A composition according to  claim 14  wherein the cytostatic or cytocidal quinone is a napthoquinone.  
     
     
         16  A composition according to  claim 15  wherein the naphthoquinone contains a hydroxy group and is linked to the peptide by said hydroxy group.  
     
     
         17  A composition according to  claim 16 , where the napthoquinone is selected from the group of compounds of the formulas  
       
         
           
           
               
               
           
         
       
       wherein A is —CH 2 —, —O—, —C(═O)—O—, or —O—C(═O)—, and M 1  is C 1 -C 8  alkyl, C 1 -C 8  branched alkyl, C 3 -C 8  cycloalkyl, or C 3 -C 8  cycloaryl.  
     
     
         18 . A composition according to  claim 16 , where the napthoquirrone is selected from the group of compounds of the formula  
       
         
           
           
               
               
           
         
         wherein x is 1 or 2; and each K is independently selected from the group consisting of H, OH, C 1 -C 8  alkyl, C 1 -C 8  alkenyl, C 1 -C 8  alkanol, C 1 -C 8  alkoxy, and  
         
           
             
             
                 
                 
             
           
         
         with the proviso that one and only one K is selected from the group consisting of OH and C 1 -C 8  alkanol, the peptide being conjugated to the terminal hydroxy group of the alcohol;  
         and where zero or two, but no more than two, vicinal K's in the molecule represent single electrons which form a pi bond, thus forming a double bond together with the existing sigma bond between the two adjacent carbons bearing the two vicinal K's.  
       
     
     
         19 . A composition according to  claim 16 , where the napthoquinone is selected from the group of compounds of the formula  
       
         
           
           
               
               
           
         
         wherein Y is selected from the group consisting of —H, —F, —Br, —Cl, and —I; and wherein G 1  is selected from the group consisting of H, C 1 -C 8  alkyl,  
         
           
             
             
                 
                 
             
           
         
          and —C(═O)—CH n X 3-n , where n is an integer from 0 to 3 and X is selected from the group consisting of F, Cl, Br, and I; and the peptide is conjugated to the quinone via the amino group bearing G1.  
       
     
     
         20 . A composition according to  claim 16 , where the napthoquinone is selected from the group of compounds of the formula  
       
         
           
           
               
               
           
         
         wherein x is 1 or 2; and each K is independently selected from the group consisting of H, C 1 -C 8  alkyl, C 1 -C 8  alkenyl, C 1 -C 8  alkoxy, and  
         
           
             
             
                 
                 
             
           
         
         and where zero or two, but no more than two, vicinal K's in the molecule represent single electrons which form a pi bond, thus forming a double bond together with the existing sigma bond between the two adjacent carbons bearing the two vicinal K's.  
       
     
     
         21 . A composition according to  claim 15  wherein the naphthoquinone contains a primary or secondary amino group and is linked to the peptide by said amino group.  
     
     
         22 . A composition according to  claim 1 , wherein the peptide substrate is a substrate of prostate specific antigen (PSA).  
     
     
         23 . A composition according to  claim 1 , wherein the peptide sequence comprises SKLQ.  
     
     
         24 . A composition according to  claim 1 , wherein the peptide sequence comprises SKLQL or SKLQ-β-alanine.  
     
     
         25 . A composition according to  claim 1 , wherein the peptide sequence comprises HSSKLQ.  
     
     
         26 . A composition according to  claim 1  wherein the peptide substrate is a substrate of cathepsin B.  
     
     
         27 . A composition according to  claim 1 , wherein the peptide sequence is X—P2—P1, where X is hydrogen, an amino-protecting group, or an amino-capping group attached to the N-terminus of P2; P2 is a hydrophobic amino acid; and P 1  is a basic or polar amino acid.  
     
     
         28 . A composition according to  claim 1 , wherein the peptide sequence is X—P2—P1—Y, where X is hydrogen, an amino-protecting group, or an amino-capping group attached to the N-terminus of P2; P2 is a hydrophobic amino acid; P1 is a basic or polar amino acid; and where Y is leucine, β-alanine, or a nonentity.  
     
     
         29 . A composition according to  claim 27 , wherein X is a 4-morpholinocarbonyl group.  
     
     
         30 . A composition according to  claim 28 , wherein X is a 4-morpholinocarbonyl group.  
     
     
         31 . A composition according to  claim 29 , wherein P2 is selected frontA the group consisting of leucine, isoleucine, valine, methionine, and phenylalanine; and P1 is selected from the group consisting of lysine, arginine, glutamine, asparagine, histidine and citrulline.  
     
     
         32 . A composition according to  claim 30 , wherein P2 is selected from the group consisting of leucine, isoleucine, valine, methionine, and phenylalanine; and P 1  is selected from the group consisting of lysine, arginine, glutamine, asparagine, histidine and citrulline.  
     
     
         33 . Acompositioncomprising the polyamine analog conjugate of  claim 1 , and a pharmaceutically acceptable excipient.  
     
     
         34 . A method of treating a disease characterized by the proliferation of prostate cells in an individual comprising administering to the individual a therapeutic amount of a polyamine analog conjugate of  claim 1 .  
     
     
         35 . The method of  claim 34 , wherein the polyamine analog is conformationally restricted.  
     
     
         36 . The method of  claim 34 , wherein the disease is prostatitis, benign prostate hyperplasia or prostate cancer.  
     
     
         37 . The method of  claim 34 , wherein the individual is a human.