US2002177715A1PendingUtilityA1

Novel autoinducer molecules and uses therefor

Priority: Aug 31, 2000Filed: Aug 31, 2001Published: Nov 28, 2002
Est. expiryAug 31, 2020(expired)· nominal 20-yr term from priority
C07D 215/233C12Q 1/045C12P 17/12C12Q 1/025C12N 1/20
40
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Claims

Abstract

Novel bacterial quinolone signal molecules and, more particularly, pseudomonas quinolone signal (“PQS”) molecules, e.g., 2-heptyl-3-hydroxy-4-quinolone, and analogs and derivatives thereof are described. Therapeutic compositions containing the molecules, and therapeutic methods, methods of for regulating gene expression, methods for identifying modulators of the autoinducer molecules, and methods of modulating quorum sensing signalling in bacteria using the compounds of the invention are also described.

Claims

exact text as granted — not AI-modified
What is claimed is:  
     
         1 . A compound of formula I  
       
         
           
           
               
               
           
         
       
       wherein: 
 R 1 -R 4  are independently H, alkyl, alkenyl, alkynyl, OH, NH 2 , SH, O—R 6 , N—R 7 R 8 , or a halogen;  
 R 5  is H, SH, OH, O—R6, or N—R 7 R 8 ;  
 R 6  is H or C 1 -C 4  alkyl;  
 R 7  and R 8  are independently H, C 1 -C 4  alkyl, O, or S;  
 X and Y are independently S, O, or N—R 9 ;  
 R 9  is H, O, S, or C 1 -C 4  alkyl;  
 Q is a tail group; and  
 salts thereof.  
 
     
     
         2 . The compound of  claim 1 , wherein Q has formula IA  
       
         
           
           
               
               
           
         
       
       wherein: 
 R 10 -R 13  are independently H, C 1 -C 4  alkyl, OH, NH 2 , SH, O—R 25 , N—R 26 R 27 , or a halogen, or R 10  and R 11  taken together form a carbonyl, a sulfonyl or an imino moiety, or R 12  and R 13  taken together form a carbonyl, a sulfonyl or an imino moiety;  
 R 14 -R 24  are independently H, C 1 -C 4  alkyl, OH, NH 2 , SH, O—R 25 , N—R 26 R 27 , or a halogen;  
 R 25  is H or C 1 -C 4  alkyl; and  
 R 26  and R 27  are independently H, C 1 -C 4  alkyl, O, or S.  
 
     
     
         3 . The compound of  claim 2  that is different than 2-heptyl-3-hydroxy-4-quinolone.  
     
     
         4 . The compound of  claim 2 , wherein R 16 , R 17 , and R 18  are H.  
     
     
         5 . The compound of  claim 2 , wherein R 2  is halogen.  
     
     
         6 . The compound of  claim 2 , wherein R 3  is halogen.  
     
     
         7 . The compound of  claim 2 , wherein R is halogen.  
     
     
         8 . The compound of  claim 2 , wherein X is S or N—R 9 .  
     
     
         9 . The compound of  claim 2 , wherein Y is O, S, or N—R 9  and wherein R 9  is C 1 -C 4 -alkyl.  
     
     
         10 . The compound of  claim 2 , wherein R 5  is H, SH, O—R 6 , or N—R 7 R 8 , and wherein R 6  is C 1 -C 4  alkyl.  
     
     
         11 . The compound of  claim 2 , wherein R 5  is SH, O—R 6 , or N—R 7 R 8 .  
     
     
         12 . The compound of  claim 2 , wherein X is O.  
     
     
         13 . The compound of  claim 12 , wherein R 5  is OH and Y is N—R 9 .  
     
     
         14 . The compound of  claim 1 , wherein Q is an alkylene chain having a skeleton of three to twenty carbon atoms.  
     
     
         15 . The compound of  claim 14 , wherein the alkylene chain contains one or more double bonds or triple bonds between the carbon atoms forming the skeleton alkylene side chain.  
     
     
         16 . The compound of  claim 14 , wherein one or more carbon atoms forming the skeleton of the alkylene side chain are replaced with sulfur or sulfur-substituted moieties.  
     
     
         17 . The compound of  claim 2 , wherein the compound contains a chiral center.  
     
     
         18 . The compound of  claim 2 , which is an optically active isomer.  
     
     
         19 . The compound of  claim 1 , comprising the formula:  
       
         
           
           
               
               
           
         
       
     
     
         20 . An autoinducer molecule comprising a compound of any one of claims  1 ,  2  or  19 .  
     
     
         21 . The autoinducer molecule of  claim 20  that regulates gene expression.  
     
     
         22 . The autoinducer molecule of  claim 21  that regulates gene expression in bacteria.  
     
     
         23 . The autoinducer molecule of  claim 22 , wherein said bacteria is  Pseudomonas aeruginosa.    
     
     
         24 . The autoinducer molecule of  claim 23 , wherein said gene expresses a virulence factor.  
     
     
         25 . The autoinducer molecule of  claim 24 , wherein the virulence factor is elastase.  
     
     
         26 . The autoinducer of  claim 20  that regulates the activity of the LasR protein of  Pseudomonas aeruginosa.    
     
     
         27 . The autoinducer of  claim 20  that regulates the activity of the RhlR protein of  Pseudomonas aeruginosa.    
     
     
         28 . The autoinducer molecule of  claim 20  that is isolated from culture media in which  Pseudomonas aeruginosa  is grown.  
     
     
         29 . A compound of claims  1  or  2  that modulates the autoinducer activity of 2-heptyl-3-hydroxy-4-quinolone.  
     
     
         30 . The compound of  claim 29  that inhibits the autoinducer activity of 2-heptyl-3-hydroxy-4-quinolone.  
     
     
         31 . The compound of  claim 29  that synergistically enhances the autoinducer activity of 2-heptyl-3-hydroxy-4-quinolone.  
     
     
         32 . A compound of claims  1  or  2  that modulates the activity of the LasR and/or the RhlR proteins of  Pseudomonas aeruginosa.    
     
     
         33 . The compound of  claim 32  that is an antagonist of the LasR and/or the RhlR proteins of  Pseudomonas aeruginosa.    
     
     
         34 . The compound of  claim 32  that is an antagonist of the LasR and/or the RhlR proteins of  Pseudomonas aeruginosa.    
     
     
         35 . A pharmaceutical composition comprising a therapeutically effective amount of a compound of  claim 1  and a pharmaceutically acceptable carrier therefor, wherein the compound inhibits the activity of one or more proteins in a microorganism that regulate expression of virulence factors.  
     
     
         36 . The pharmaceutical composition of  claim 35 , wherein the compound is present in an amount effective to affect the ability of the microorganism to initially infect or further infect an organism.  
     
     
         37 . The pharmaceutical composition of  claim 35 , wherein the microorganism is  Pseudomonas aeruginosa.    
     
     
         38 . The pharmaceutical composition of  claim 37 , wherein the compound inhibits the activity of the LasR and/or the RhlR proteins of  Pseudomonas aeruginosa.    
     
     
         39 . The pharmaceutical composition of  claim 38 , wherein the compound inhibits the autoinducer activity of 2-heptyl-3-hydroxy-4-quinolone.  
     
     
         40 . The pharmaceutical composition of  claim 35 , further comprising an antimicrobial, antibacterial or antifungal agent.  
     
     
         41 . A method of inhibiting the infectivity of  Pseudomonas aeruginosa  comprising administering to a subject a therapeutically effective amount of a compound of  claim 1 , wherein the compound inhibits the activity of the LasR and/or the RhlR proteins of  Pseudomonas aeruginosa.    
     
     
         42 . The method of  claim 41 , wherein the compound inhibits the autoinducer activity of 2-heptyl-3-hydroxy-4-quinolone.  
     
     
         43 . A method of treating an immunocompromised subject infected with  Pseudomonas aeruginosa  comprising administering to a subject a therapeutically effective amount of a compound of  claim 1 , wherein the compound inhibits the activity of the LasR and/or the RhlR proteins of  Pseudomonas aeruginosa.    
     
     
         44 . The method of  claim 43 , wherein the compound inhibits the autoinducer activity of 2-heptyl-3-hydroxy-4-quinolone.  
     
     
         45 . The method of  claim 43 , wherein the subject is afflicted with cystic fibrosis.  
     
     
         46 . A culture medium for microorganisms comprising, as an added compound, an autoinducer molecule as defined in  claim 20 , at a concentration effective to stimulate or promote the metabolism, growth and/or recovery of the microorganism.  
     
     
         47 . The culture medium of  claim 46 , wherein the microorganism is  Pseudomonas aeruginosa.    
     
     
         48 . The culture medium of  claim 47 , wherein the autoinducer is 2-heptyl-3-hydroxy-4-quinolone.  
     
     
         49 . A method for identifying a compound that modulates an autoinducer molecule in bacteria, said method comprising: 
 providing a cell which comprises a quorum sensing controlled gene, wherein said cell is responsive to an autoinducer molecule of  claim 20  such that a detectable signal is generated;    contacting said cell with an autoinducer as defined in  claim 20  in the presence and absence of a test compound; and    detecting a change in the detectable signal to thereby identify said test compound as a modulator of an autoinducer molecule in bacteria.    
     
     
         50 . The method of  claim 49 , wherein the compound inhibits the autoinducer molecule.  
     
     
         51 . The method of  claim 49 , wherein the compound synergizes activity of the autoinducer molecule.  
     
     
         52 . The method of  claim 49 , wherein said bacteria is  Pseudomonas aeruginosa.    
     
     
         53 . The method of  claim 49 , wherein the autoinducer is 2-heptyl-3-hydroxy-4-quinolone.  
     
     
         54 . The method of  claim 52 , wherein the compound inhibits binding of the autoinducer molecule to LasR and/or RhlR.  
     
     
         55 . A method of regulating the expression of a gene in bacteria comprising: 
 inserting a gene into bacteria chosen for enhancement of gene expression by a compound of  claim 1  that enhances the activity of the LasR and/or RhlR protein; and    incubating the bacteria with a compound of  claim 1  that enhances the activity of the LasR protein, such that the expression of the gene is regulated.    
     
     
         56 . The method of  claim 55  wherein the method further comprises the additional steps of: 
 allowing the gene expression to reach a desired level; and  
 incubating the bacteria with a compound of  claim 1  that inhibits the activity of the LasR and/or RhlR protein, thereby regulating the gene expression by the bacteria.  
 
     
     
         57 . An inhibitor of the autoinducer activity of 2-heptyl-3-hydroxy-4-quinolone.  
     
     
         58 . An analog of 2-heptyl-3-hydroxy-4-quinolone that inhibits the induction of virulence factors by 2-heptyl-3-hydroxy-4-quinolone, LasR or RhlR.  
     
     
         59 . The analog of  claim 58 , wherein the virulence factor is exotoxin A.  
     
     
         60 . The analog of  claim 58 , wherein the virulence factor is elastase.  
     
     
         61 . The analog of  claim 58 , wherein the virulence factor is an alkaline protease.  
     
     
         62 . An analog of 2-heptyl-3-hydroxy-4-quinolone that inhibits the induction of biofilm formation by 2-heptyl-3-hydroxy-4-quinolone, LasR or RhlR.  
     
     
         63 . A method for modulating quorum sensing signaling in bacteria, said method comprising: 
 providing bacteria that comprise a quorum sensing controlled gene, wherein said bacteria are responsive to an autoinducer molecule; and    incubating the bacteria with a compound of  claim 3 , such that quorum sensing signalling in bacteria is modulated.    
     
     
         64 . The method of claim  63 , wherein the autoinducer molecule is 2-heptyl-3-hydroxy-4-quinolone.

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