Materials and methods for treating diseases caused by genetic disorders using aminoglycosides and derivatives thereof which exhibit low nephrotoxicity

Abstract

Various aspects related to the preparation of congeners of the aminoglycosides gentamicin such as the congener C2 and using this compound or derivatives thereof and pharmaceutically active salts to treat diseases that involve genetic mutations which introduce a missense or premature stop codon into a gene. Still other aspects include treating human or animal patients with the gentamicin congener C2 and derivatives and pharmaceutical salt thereof to overcome, or to at least mitigate, the symptoms of disease and disorders such as some forms of Becker's or Duchenne muscular dystrophy, Hurler's Syndrome and Cystic Fibrosis that have as their etiology the presence of a premature stop codon in a gene whose proper expression is necessary for good health.

1 . A composition for promoting readthrough of a stop codon, comprising: 
     an isolated and purified compound having the following formula, or a pharmaceutically acceptable salt thereof: 
     
       
         
         
             
             
         
       
     
     wherein R 1  and R 3  are H, and R 2  is CH 3 . 
   
   
       2 . The compound according to  claim 1 , wherein said compound is suitable for promoting readthrough of a stop codon selected from the group consisting of: UAA, UGA and UAG. 
   
   
       3 . The compound according to  claim 1 , wherein said compound is suitable for promoting readthrough of a stop codon having the sequence UAG. 
   
   
       4 . A method for promoting readthrough of a stop codon, comprising the steps of:
 identifying a nucleic acid included in a system for RNA translation, wherein said nucleic acid includes a stop codon; and   providing an isolated and purified compound having the following formula, or a pharmaceutically acceptable salt thereof:   
     
       
         
         
             
             
         
       
     
     wherein R 1  and R 3  are H, and R 2  is CH 3 ; and 
     administering said compound in an in an amount sufficient to promote readthrough of the stop codon in the nucleic acid. 
   
   
       5 . A method of treating a genetic disease or disorder, comprising the steps of:
 identifying a patient, wherein the patient has a pathology including the imposition of a stop codon in the open reading frame of an otherwise functional gene; and   administering a therapeutically effective dose of an isolated and purified compound having the following formula, or a pharmaceutically acceptable salt thereof:   
     
       
         
         
             
             
         
       
     
     wherein R 1  and R 3  are H, and R 2  is CH 3 . 
   
   
       6 . The method described in  claim 5 , wherein the genetic disease is selected from the group consisting of: some forms of Duchenne Muscular Dystrophy, cystic fibrosis, Hurler's Syndrome, and Becker muscular dystrophies. 
   
   
       7 . The method described in  claim 3 , wherein the patient is a human. 
   
   
       8 . A method of treating a genetic disease, comprising the steps of:
 identifying a patient, wherein the patient exhibits a pathology that includes the imposition of a stop codon in the open reading frame of an otherwise functional gene; and   formulating a mixture of a therapeutically effective dose of an aminoglyocide enriched in an isolated and purified compound having the following formula, or a pharmaceutically acceptable salt thereof:   
     
       
         
         
             
             
         
       
     
     wherein, R 1  and R 3  are H, and R 2  is CH 3 , and wherein the compound is present in said formulation at a level sufficient to reduce the cellular toxicity of the aminoglyoside; and 
     administering a therapeutically effective dose of said formulation to the patient. 
   
   
       9 . A kit for promoting readthrough of a stop codon, comprising: 
     an isolated and purified compound having the following formula: 
     
       
         
         
             
             
         
       
     
     wherein, R 1  and R 3  are H, and R 2  is CH 3 . 
   
   
       10 . The method described in any of  claims 1 ,  4 ,  5 ,  6 ,  7 ,  8  and  9 , wherein the stop codon has the sequence UAGC. 
   
   
       11 . The method described in any of  claims 1 ,  4 ,  5 ,  6 ,  7 ,  8 , and  9 , wherein the stop codon is selected from the following consisting of the following sequences: UAAC, UAGC, UGAC, UAAG, UAGG, UGAG, UAAU, UAGU, UGAU, UAAA, UAGA, and UGAA.