Screening molecules with anti-prion activity: kits, methods and screened molecules

Abstract

A kit and a method for identifying compounds having anti-prion activity are provided. The kit comprises a yeast of phenotype [PSI+]; an antibiogram; and a prion curing agent in a sub-effective dose, wherein the yeast has the adel-14 allele of the ADE1 gene and an inactivated ERG6 gene. Compounds and pharmaceutical compositions having anti-prion activity are also provided, which are useful for treating various neurodegenerative diseases, including polyglutamines expansion associated diseases; Huntington's disease; Kennedy disease; amyotrophic lateral sclerosis; cerebellous autosomic ataxies; dentalorubral-pallidoluysian atrophy; and spino-bulbar amyotrophy.

1 . A kit for screening molecules having an anti-prion activity, comprising: 
 a yeast of phenotype [PSI+];    an antibiogram; and    a prion curing agent in a sub-effective dose,    wherein the yeast has the adel-14 allele of the ADE1 gene and an inactivated ERG6 gene.    
     
     
         2 . The kit of  claim 1 , wherein the yeast is  Saccharomyces cerevisiae.    
     
     
         3 . The kit of  claim 1 , wherein the prion curing agent is guanidium chloride.  
     
     
         4 . A method for screening molecules having anti-prion activity, the method comprising: 
 a. producing in vitro a lawn of cells on a medium containing a sub-effective dose of a prion curing agent;    b. contacting the cells with a test compound according to the antibiogram method;    c. incubating the cells for approximately 2-4 days at approximately 20-25° C.; and    d. evaluating the staining of the cell colonies,    wherein the cells comprise yeasts of [PSI+] phenotype having the adel-14 allele of the ADE1 gene and an inactivated ERG6 gene.    
     
     
         5 . The screening method of  claim 4 , wherein the yeast is  Saccharomyces cerevisiae.    
     
     
         6 . The screening method of  claim 4 , wherein the curing agent is guanidium chloride.  
     
     
         7 . The screening method of  claim 4  further comprising: 
 e. incubating for approximately 2-4 days at approximately 2-6° C.; and/or    f. carrying out a secondary screening test.    
     
     
         8 . The screening method of  claim 7 , wherein the secondary screening test comprises: 
 constructing a strain of yeast in which the ADE2 gene is under the control of the DAL5 gene promoter;    producing in vitro a lawn of cells on a medium containing a sub-effective dose of a prion curing agent;    contacting the cells with a test compound according to the antibiogram method;    incubating the cells for approximately 2-4 days at approximately 20-25° C.;    evaluating the staining of the cell colonies; and    incubating for approximately 2-4 days at approximately 2-6° C.    
     
     
         9 . A medicament comprising the compound of formula (II):  
       
         
           
           
               
               
           
         
       
       wherein R′represents an H, NH 2 , NH—(CH 2 ) 3 —N(CH 3 ) 2 , NH—CH(CH 3 )—(CH 2 ) 3 —N(CH 2 —CH 3 ) 2 , 
 X represents F, Cl , CF 3 ,  
 p and n, identical or different, are equal to 0, 1 or 2.  
 
     
     
         10 . The medicament of  claim 9  comprising the compound of formula (II):  
       
         
           
           
               
               
           
         
       
       wherein R′represents an NH 2  group, 
 X represents F, C 1 , CF 3 ,  
 p and n, identical or different, are equal to 0, 1 or 2.  
 
     
     
         11 . A method for treating neurodegenerative diseases involving protein aggregates, the method comprising: 
 administering the compound of formula (I)                          wherein R′ is an H, NH 2 , NHR 2  group, where R 2  is an alkyl or alkylaminoalkyl chain with 1 to 10 carbon atoms, branched or unbranched,    X represents F, Cl, Br, I, CF 3 , SCH 3 , OCH 3 , OH, NO 2 , COCH 3 , CONH 2 , COOH, COOR 3 , where R 3  is an alkyl group with 1 to 4 carbon atoms,    p and n, identical or different, are equal to 0, 1 or 2,    q is equal to 0 or 1.    
     
     
         12 . A method for treating neurodegenerative diseases involving protein aggregates, the method comprising:  
       
         
           
           
               
               
           
         
         administering the compound of formula (III)  
         wherein R′ represents an H, NH 2 , NH—(CH 2 ) 3 —N(CH 3 ) 2 , NH—CH(CH 3 )—(CH 2 ) 3 —N(CH 2 —CH 3 ) 2  group,  
         X represents F, Cl, CF 3 ,  
         p and n, identical or different, are equal to 0, 1 or 2.  
       
     
     
         13 . A method for treating neurodegenerative diseases involving protein aggregates, the method comprising: 
 administering the compound of formula (II)                          wherein R represents an H, NH 2 , NH—(CH 2 ) 3 —N(CH 3 ) 2 , NH—CH(CH 3 )—(CH 2 ) 3 —N(CH 2 —CH 3 ) 2  group,    X represents F, Cl, CF 3 ,    p and n, identical or different, are equal to 0, 1 or 2.    
     
     
         14 . A method for treating neurodegenerative diseases involving protein aggregates, the method comprising: 
 administering the compound of formula (II)                          wherein R′represents an NH 2  group,    X represents F, Cl, CF 3 ,    p and n, identical or different, are equal to 0, 1 or 2.    
     
     
         15 . The method of  claim 11 , wherein the neurodegenerative diseases include:  
       spongiform encephalopathies, Alzheimer's disease, and Huntington's disease.  
     
     
         16 . The method of  claim 12 , wherein the neurodegenerative diseases include:  
       spongiform encephalopathies, Alzheimer's disease, and Huntington's disease.  
     
     
         17 . The method of  claim 13 , wherein the neurodegenerative diseases include:  
       spongiform encephalopathies, Alzheimer's disease, and Huntington's disease.  
     
     
         18 . The method of  claim 13 , wherein the neurodegenerative diseases include: 
 polyglutamines expansion associated diseases;    Huntington's disease;    Kennedy disease;    the amyotrophic lateral sclerosis;    cerebellous autosomic ataxies;    dentalorubral-pallidoluysian atrophy; and    spino-bulbar amyotrophy.    
     
     
         19 . A pharmaceutical composition comprising: 
 a therapeutically effective quantity of at least one compound of formula (II)                          wherein R′ represents an H, NH 2 , NH—(CH 2 ) 3 —N(CH 3 ) 2 , NH—CH(CH 3 )—(CH 2 ) 3 —N(CH 2 —CH 3 ) 2  group,    X represents F, Cl, CF 3 ,    p and n, identical or different, are equal to 0, 1 or 2,    in combination with at least one pharmaceutically acceptable vehicle.    
     
     
         20 . A pharmaceutical composition comprising: 
 a therapeutically effective quantity of at least one compound of formula (II)                          wherein R′represents an NH 2  group,    X represents F, Cl, CF 3 ,    p and n, identical or different, are equal to 0, 1 or 2,    in combination with at least one pharmaceutically acceptable vehicle.