Method for reducing or preventing the establishment, growth or metastasis of cancer by administering indole peptidomimetics PAR-1 antagonist and optionally PAR-2 antagonists

Abstract

We have discovered a method of modifying the tumor cell microenvironment to reduce or prevent the establishment, growth or metastasis of malignant cells comprising administering to a patient having malignant cells a pharmaceutically effective amount of a PAR-1 inhibitor and optionally a PAR-2 inhibitor to prevent or reduce activation of normal cells within the tumor microenvironment. This method also has the effect in some patients of modulating the immune system to facilitate a more efficient immune response to malignant cells and maybe coupled with cytokine therapy and T-cell therapy to enhance the patient's immune response to the malignant cells.

We claim:  
     
         1 . A method of modifying the tumor cell microenvironment to reduce or prevent the establishment, growth or metastasis of malignant cells that directly or indirectly activate the PAR-1 receptor of normal cells comprising providing a pharmaceutically effective amount of a PAR-1 inhibitor of the following formula (I):  
       
         
           
           
               
               
           
         
       
       wherein: 
 A 1  and A 2  are each independently a D- or L-amino acid selected from the group consisting of alanine, β-alanine, arginine, homoarginine, cyclohexylalanine, citrulline, cysteine (optionally substituted with C 1 -C 4  alkyl, aryl, or arC 1 -C 4  alkyl), 2,4-diaminobutyric acid (optionally substituted with acyl, C 1 -C 4  alkyl, aroyl, amidino, or MeC(NH)—), 2,3-diaminopropionic acid (optionally substituted with acyl, C 1 -C 4  alkyl, aroyl, amidino, or MeC(NH)—), glutamine, glycine, indanylglycine, lysine (optionally substituted with acyl, C 1 -C 4  alkyl, aroyl, MeC(NH)—), valine, methionine, proline, serine (optionally substituted with C 1 -C 4  alkyl, aryl, or arC 1 -C 4  alkyl), homoserine (optionally substituted with C 1 -C 4  alkyl, aryl, or arC 1 -C 4  alkyl), tetrahydroisoquinoline-3-COOH, threonine (optionally substituted with C 1 -C 4  alkyl, aryl, or arC 1 -C 4  alkyl), omithine (optionally substituted with acyl, C 1 -C 4  alkyl, aroyl, MeC(NH)—), and an unsubstituted or substituted aromatic amino acid selected from the group consisting of phenylalanine, heteroarylalanine, naphthylalanine, homophenylalanine, histidine, tryptophan, tyrosine, arylglycine, heteroarylglycine, aryl-β-alanine, and heteroaryl-β-alanine wherein the substituents on the aromatic amino acid are independently selected from one or more of halogen, C 1 -C 4  alkyl, C 1 -C 4  alkoxy, hydroxy, C 1 -C 4  alkoxycarbonyl, amino, amidino, guanidino, fluorinated C 1 -C 4  alkyl, fluorinated C 1 -C 4  alkoxy, C 1 -C 4  alkylsulfonyl, C 1 -C 4  alkylcarbonyl, cyano, aryl, heteroaryl, arC 1 -C 4  alkyl, C 2 -C 4  alkenyl, alkynyl, or nitro;  
 R 1  is selected from amino, C 1 -C 8  alkylamino, C 1 -C 8  dialkylamino, arylamino, arC 1 -C 8  alkylamino, C 3 -C 8  cycloalkylamino, heteroalkylC 1 -C 8  alkylamino, heteroalkylC 1 -C 8  alkyl-N-methylamino, C 1 -C 8  dialkylaminoC 1 -C 8  alkylamino, -N(C 1 -C 8 -alkyl-C 1 -C 8  alkyl-N(C 1 -C 8 alkyl) 2 , N(C 1 -C 8  alkyl)(C 1 -C 8  alkenyl), -N(C 1 -C 8 alkyl)(C 3 -C 8 cycloalkyl), heteroalkyl or substituted heteroalkyl wherein the substituent on the heteroalkyl is selected from oxo, amino, C 1 -C 8  alkoxyC 1 -C 8  alkyl, C 1 -C 8  alkylamino or C 1 -C 8  dialkylamino;  
 R 2  is selected from hydrogen, halogen, C 1 -C 8  alkyl, C 3 -C 8  cycloalkyl, C 1 -C 8  alkenyl, C 1 -C 8  alkynyl, arC 1 -C 8  alkyl, aryl or heteroaryl;  
 R 3  and R 4  are each independently selected from hydrogen, C 1 -C 8  alkyl, C 3 -C 8  cycloalkyl, C 3 -C 8  cycloalkylC 1 -C 8  alkyl, aryl, heteroalkyl, substituted heteroalkyl (wherein the substituent on the heteroalkyl is one or more substituents independently selected from C 1 -C 8  alkoxycarbonyl, C 1 -C 8  alkyl, or C 1 -C 4  alkylcarbonyl), heteroalkylci-C 8  alkyl, indanyl, acetamidinoC 1 -C 8  alkyl, aminoC 1 -C 8  alkyl, C 1 -C 8  alkylaminoC 1 -C 8  alkyl, C 1 -C 8  dialkylaminoC 1 -C 8  alkyl, unsubstituted or substituted heteroarylC 1 -C 8  alkyl, or unsubstituted or substituted arC 1 -C 8  alkyl, wherein the substituent on the aralkyl or heteroarylalkyl group is one or more substituents independently selected from halogen, nitro, amino, C 1 -C 8  alkyl, C 1 -C 8  alkoxy, hydroxy, cyano, C 1 -C 4  alkylcarbonyl, C 1 -C 8  alkoxycarbonyl, hydroxyC 1 -C 8  alkyl or aminosulfonyl; or  
 R 3  and R 4 , together with the nitrogen to which they are attached, alternatively form an unsubstituted or substituted heteroalkyl group selected from piperidinyl, piperazinyl, morpholinyl or pyrrolidinyl, wherein the substituent is one or more substituents independently selected from C 1 -C 8  alkyl C 1 -C 8  alkoxycarbonyl or C 1 -C 4  alkylcarbonyl;  
 R 5  is selected from unsubstituted or substituted aryl, arC 1 -C 8  alkyl, C 3 -C 8  cycloalkyl, or heteroaryl, where the substituents on the aryl, arC 1 -C 8  alkyl, cycloalkyl or heteroaryl group are independently selected from one or more of halogen, nitro, amino, cyano, hydroxyalkyl, C 1 -C 8  alkyl, C 1 -C 8  alkoxy, hydroxy, C 1 -C 4  alkylcarbonyl, C 1 -C 8  alkoxycarbonyl, fluorinated C 1 -C 4  alkyl, fluorinated C 1 -C 4  alkoxy or C 1 -C 4  alkylsulfonyl;  
 R 6  is selected from hydrogen or C 1 -C 8  alkyl,  
 X is oxygen or sulfur;  
 m is an integer selected from 0, 1, 2 or 3;  
 n is an integer selected from 1 or 2; and  
 p is an integer selected from 0 or 1;  
 and pharmaceutically acceptable salts thereof and optionally a PAR-2 inhibitor to a patient with malignant cells that directly or indirectly activate PAR-1 and/or PAR-2.  
 
     
     
         2 . The method of  claim 1  wherein the PAR-1 inhibitor is a compounds of the formula:  
       
         
           
           
               
               
           
         
       
       wherein: 
 A 1  is an L-amino acid selected from the group consisting of alanine, arginine, cyclohexylalanine, proline, tetrahydroisoquinoline-3-COOH, and an unsubstituted or substituted aromatic amino acid selected from the group consisting of phenylalanine, naphthylalanine, homophenylalanine, and O-methyl tyrosine, wherein the substituents on the aromatic amino acid are independently one to two substituents selected from halogen, C 1 -C 4  alkyl, C 1 -C 4  alkoxy, hydroxy, C 1 -C 4  alkoxycarbonyl, amino, amidino, guanidino, fluorinated C 1 -C 4  alkyl, fluorinated C 1 -C 4  alkoxy, C 1 -C 4  alkylsulfonyl, C 1 -C 4  alkylcarbonyl, cyano, aryl, heteroaryl, arC 1 -C 4  alkyl, C 2 -C 4  alkenyl, alkynyl, or nitro;  
 A 2  is an L-amino acid selected from the group consisting of alanine, alanine, arginine, citrulline, cysteine (optionally substituted with C 1 -C 4  alkyl, aryl, or arC 1 -C 4  alkyl), 2,4-diaminobutyric acid (optionally substituted with acyl, C 1 -C 4  alkyl, aroyl, amidino, or MeC(NH)—), 2,3-diaminopropionic acid (optionally substituted with acyl, C 1 -C 4  alkyl, aroyl, amidino, or MeC(NH)—), glutamine, glycine, lysine (optionally substituted with acyl, C 1 -C 4  alkyl, aroyl, MeC(NH)—), valine, methionine, serine (optionally substituted with C 1 -C 4  alkyl, aryl, or arC 1 -C 4  alkyl), homoserine (optionally substituted with C 1 -C 4  alkyl, aryl, or arC 1 -C 4  alkyl), threonine (optionally substituted with C 1 -C 4  alkyl, aryl, or arC 1 -C 4  alkyl), ornithine (optionally substituted with acyl, C 1 -C 4  alkyl, aroyl, MeC(NH)—), and an unsubstituted or substituted aromatic amino acid selected from the group consisting of phenylalanine, heteroarylalanine, and histidine, wherein the substituents on the aromatic amino acid are independently one to two substituents selected from halogen, C 1 -C 4  alkyl, C 1 -C 4  alkoxy, hydroxy, C 1 -C 4  alkoxycarbonyl, amino, amidino, guanidino, fluorinated C 1 -C 4  alkyl, fluorinated C 1 -C 4  alkoxy, C 1 -C 4  alkylsulfonyl, C 1 -C 4  alkylcarbonyl, cyano, aryl, heteroaryl, arC 1 -C 4  alkyl, C 2 -C 4  alkenyl, alkynyl, or nitro;  
 R 1  is selected from diethylamino, di-(n-propyl)amino,  
                     
 R 3  is selected from hydrogen, methyl or ethyl;  
 R 4  is selected from 2-indanyl, phenyl, cyclohexylmethyl, cyclopentyl, pyridylmethyl, furanylmenthyl, 2-(4-methyl-furanyl)methyl, thienylmethyl, diphenylmethyl, 4-imidazolylethyl, 2-(4-N-methyl)imidazolylethyl, n-octyl, phenyl-n-propyl, aminoethyl, aminopropyl, amino-n-pentyl, dimethylaminoethyl, 4-aminophenylsulfonylarninomethyl, acetamidineylethyl, 2-N-pyrrolidinylethyl, N-ethoxycarbonylpiperidinyl, unsubstituted or substituted phenylethyl and unsubstituted or substituted benzyl wherein the substituents on the phenylethyl or benzyl are independently one or two substituents selected from methyl, fluorine, chlorine, nitro, methoxy, methoxycarbonyl or hydroxymethyl; or  
 R 3  and R 4 , together with the nitrogen to which they are attached, alternatively form a heteroalkyl group selected from piperidinyl or 4-(N-methyl)piperazinyl; and  
 R 5  is selected from cyclohexyl, 2-naphthyl, phenylethyl, 4-fluorophenylethyl, or unsubstituted or substituted phenyl, where the substituents on the phenyl are independently selected from one to two substituents selected from fluorine, chlorine, iodine, methyl, cyano or trifluoromethyl;  
 and pharmaceutically acceptable salts thereof.  
 
     
     
         3 . The method of  claim 1  wherein the PAR-1 inhibitor is administered with a therapeutically effective amount of at least one PAR-2 inhibitor.  
     
     
         4 . The method of  claim 2  wherein the PAR-1 inhibitor is administered with a therapeutically effective amount of a cytokine selected from the group consisting of IL-2, IL-12, IL-18, G-CSF, M-CSF, GM-CSF, INF-α, INF-β, INF-γ, TNF and combinations thereof.  
     
     
         5 . The method of  claim 4  wherein additionally administered in a pharmaceutical effective amount is at least one conventional chemotherapy agent.  
     
     
         6 . The method of  claim 5  wherein the chemotherapy agent is selected from the group consisting of antiangiogenic compounds, alkylating compounds, antimetabolites, hormonal agonist/antagonists, monoclonal antibodies for cancer treatment, antiproliferative compounds and combinations thereof.  
     
     
         7 . The method of  claim 2  wherein additional administered are T cells selected from the group consisting of activated T cells, activated NK cells and combinations thereof.  
     
     
         8 . The method of  claim 1  wherein the PAR-1 inhibitor is administered before surgery.  
     
     
         9 . The method of  claim 1  wherein the PAR-1 inhibitor is administered after surgery.  
     
     
         10 . A method for the modulation of the immune system to enhance a patient's immune response to malignant cells that directly or indirectly activate the PAR-1 receptor of normal cells comprising administer a pharmaceutically effective dose of a PAR-1 inhibitor of the following formula (I):  
       
         
           
           
               
               
           
         
       
       wherein: 
 A 1  and A 2  are each independently a D- or L-amino acid selected from the group consisting of alanine, β-alanine, arginine, homoarginine, cyclohexylalanine, citrulline, cysteine (optionally substituted with C 1 -C 4  alkyl, aryl, or arC 1 -C 4  alkyl), 2,4-diaminobutyric acid (optionally substituted with acyl, C 1 -C 4  alkyl, aroyl, amidino, or MeC(NH)—), 2,3-diaminopropionic acid (optionally substituted with acyl, C 1 -C 4  alkyl, aroyl, amidino, or MeC(NH)—), glutamine, glycine, indanylglycine, lysine (optionally substituted with acyl, C 1 -C 4  alkyl, aroyl, MeC(NH)—), valine, methionine, proline, serine (optionally substituted with C 1 -C 4  alkyl, aryl, or arC 1 -C 4  alkyl), homoserine (optionally substituted with C 1 -C 4  alkyl, aryl, or arC 1 -C 4  alkyl), tetrahydroisoquinoline-3-COOH, threonine (optionally substituted with C 1 -C 4  alkyl, aryl, or arC 1 -C 4  alkyl), omithine (optionally substituted with acyl, C 1 -C 4  alkyl, aroyl, MeC(NH)—), and an unsubstituted or substituted aromatic amino acid selected from the group consisting of phenylalanine, heteroarylalanine, naphthylalanine, homophenylalanine, histidine, tryptophan, tyrosine, arylglycine, heteroarylglycine, aryl-β-alanine, and heteroaryl-β-alanine wherein the substituents on the aromatic amino acid are independently selected from one or more of halogen, C 1 -C 4  alkyl, C 1 -C 4  alkoxy, hydroxy, C 1 -C 4  alkoxycarbonyl, amino, amidino, guanidino, fluorinated C 1 -C 4  alkyl, fluorinated C 1 -C 4  alkoxy, C 1 -C 4  alkylsulfonyl, C 1 -C 4  alkylcarbonyl, cyano, aryl, heteroaryl, arC 1 -C 4  alkyl, C 2 -C 4  alkenyl, alkynyl, or nitro;  
 R 1  is selected from amino, C 1 -C 8  alkylamino, C 1 -C 8  dialkylamino, arylamino, arC 1 -C 8  alkylamino, C 3 -C 8  cycloalkylamino, heteroalkylC 1 -C 8  alkylamino, heteroalkylC 1 -C 8  alkyl-N-methylamino, C 1 -C 8  dialkylaminoC 1 -C 8  alkylamino, —N(C 1 -C 8 alkyl)-C 1 -C 8  alkyl-N(C 1 -C 8 alkyl) 2 , N(C 1 -C 8  alkyl(C 1 -C 8  alkenyl), —N(C 1 -C 8 alkyl)(C 3 -C 8 cycloalkly), heteroalkyl or substituted heteroalkyl wherein the substituent on the heteroalkyl is selected from oxo, amino, C 1 -C 8  alkoxyC 1 -C 8  alkyl, C 1 -C 8  alkylamino or C 1 -C 8  dialkylamino;  
 R 2  is selected from hydrogen, halogen, C 1 -C 8  alkyl, C 3 -C 8  cycloalkyl, C 1 -C 8  alkenyl, C 1 -C 8  alkynyl, arC 1 -C 8  alkyl, aryl or heteroaryl;  
 R 3  and R 4  are each independently selected from hydrogen, C 1 -C 8  alkyl, C 3 -C 8  cycloalkyl, C 3 -C 8  cycloalkylC 1 -C 8  alkyl, aryl, heteroalkyl, substituted heteroalkyl (wherein the substituent on the heteroalkyl is one or more substituents independently selected from C 1 -C 8  alkoxycarbonyl, C 1 -C 8  alkyl, or C 1 -C 4  alkylcarbonyl), heteroalkylC 1 -C 8  alkyl, indanyl, acetamidinoC 1 -C 8  alkyl, aminoC 1 -C 8  alkyl, C 1 -C 8  alkylamino C 1 -C 8  alkyl, C 1 -C 8  dialkylaminoC 1 -C 8  alkyl, unsubstituted or substituted heteroarylC 1 -C 8  alkyl, or unsubstituted or substituted arC 1 -C 8  alkyl, wherein the substituent on the aralkyl or heteroarylalkyl group is one or more substituents independently selected from halogen, nitro, amino, C 1 -C 8  alkyl, C 1 -C 8  alkoxy, hydroxy, cyano, C 1 -C 4  alkylcarbonyl, C 1 -C 8  alkoxycarbonyl, hydroxy C 1 -C 8  alkyl or aminosulfonyl; or  
 R 3  and R 4 , together with the nitrogen to which they are attached, alternatively form an unsubstituted or substituted heteroalkyl group selected from piperidinyl, piperazinyl, morpholinyl or pyrrolidinyl, wherein the substituent is one or more substituents independently selected from C 1 -C 8  alkyl C 1 -C 8  alkoxycarbonyl or C 1 -C 4  alkylcarbonyl;  
 R 5  is selected from unsubstituted or substituted aryl, arC 1 -C 8  alkyl, C 3 -C 8  cycloalkyl, or heteroaryl, where the substituents on the aryl, arC 1 -C 8  alkyl, cycloalkyl or heteroaryl group are independently selected from one or more of halogen, nitro, amino, cyano, hydroxyalkyl, C 1 -C 8  alkyl, C 1 -C 8  alkoxy, hydroxy, C 1 -C 4  alkylcarbonyl, C 1 -C 8  alkoxycarbonyl, fluorinated C 1 -C 4  alkyl, fluorinated C 1 -C 4  alkoxy or C 1 -C 4  alkylsulfonyl;  
 R 6  is selected from hydrogen or C 1 -C 8  alkyl,  
 X is oxygen or sulfur;  
 m is an integer selected from 0, 1, 2 or 3;  
 n is an integer selected from 1 or 2; and  
 p is an integer selected from 0 or 1;  
 and pharmaceutically acceptable salts thereof and optionally a PAR-2 inhibitor to a patient to enhance the patient's immune response to the malignant cells.  
 
     
     
         11 . The method of  claim 10  wherein additionally administered are cytokines to facilitate the development of a Th1 response.  
     
     
         12 . The method of  claim 11  wherein the cytokines are selected from the group consisting of IL-2, IL12, IL-18, INF-α, INF-β, INF-γ, TNF and combinations thereof.  
     
     
         13 . The method of  claim 10  wherein additionally administered are T cells selected from the group consisting of activated CTL cells, activated NK cells and combinations thereof.  
     
     
         14 . The method of  claim 10  wherein additionally administered are activated NK cells.  
     
     
         15 . The method of  claim 10  wherein additionally administered are activated CTL cells.  
     
     
         16 . The method of  claim 10  wherein the PAR-1 inhibitor is administered before surgery.  
     
     
         17 . The method of  claim 10  wherein the PAR-1 inhibitor is administered after surgery.