Moenomycin biosynthesis-related compositions and methods of use thereof

Abstract

The methods and compositions described herein relate to the identification, isolation, and characterization of genes which encode proteins useful for the biosynthesis of transglycosylase inhibitors such as moes. The methods and compositions also relate to the production of such proteins, and their use in the synthesis of moes, the expression of moes, and the production of modified moes.

1 - 11 . (canceled) 
     
     
         12 . A method of synthesizing a moenomycin, a moenomycin derivative, or a moenomycin intermediate wholly or partially in vitro comprising: reacting a one or more moenomycin precursor, derivative and/or moenomycin intermediate with a one or more polypeptide selected from the group consisting of: moeA4, moeB4, moeC4, moeB5, moe A5, moeD5, moeJ5, moeE5, moeF5, moeH5, moeK5, moeM5, moeN5, moe05, moeX5, moeP5, moeR5, moeS5, moeGT1, moeGT2, moeGT3, moeGT4, and moeGT5, under conditions wherein the moenomycin, the moenomycin derivative, or the intermediate is wholly or partially synthesized. 
     
     
         13 . A method of modifying a moenomycin wholly or partially in vitro comprising: reacting a moenomycin, a moenomycin derivative or a moe intermediate with a one or more polypeptide selected from the group consisting of: moeA4, moeB4, moeC4, moeB5, moe A5, moeD5, moeJ5, moeE5, moeF5, moeH5, moeK5, moeM5, moeN5, moeO5, moeX5, moeP5, moeR5, moeS5, moeGT1, moeGT2, moeGT3, moeGT4, and moeGT5, under conditions wherein the moenomycin, the moenomycin derivative, or the moenomycin intermediate is modified. 
     
     
         14 - 15 . (canceled) 
     
     
         16 . A moenomycin derivative having the structure: 
       
         
           
           
               
               
           
         
         wherein 
         Ac is acetyl; 
         R and R 1  independently are selected from the group consisting of hydroxyl, and —NHR 2  where R 2  is hydrogen, alkyl, cycloalkyl, or substituted cycloalkyl; 
         X is hydrogen, or 
       
       
         
           
           
               
               
           
         
         R 3  is selected from the group consisting of hydrogen and hydroxyl; and 
         X 1  is selected from the group consisting of hydrogen, 
       
       
         
           
           
               
               
           
         
         R 4  is selected from the group consisting of hydrogen and hydroxyl; 
         R 5  is selected from the group consisting of hydroxyl and —NHR 6  where R 6  is selected from the group consisting of hydrogen, alkyl, cycloalkyl, and substituted cycloalkyl, and 
         R 7  is hydrogen or methyl, 
         or a pharmaceutically acceptable salt, tautomer, and/or ester thereof. 
       
     
     
         17 . The moenomycin derivative of  claim 16 , wherein R and R 1  independently are —NH 2 . 
     
     
         18 . The moenomycin derivative of  claim 16 , wherein X is hydrogen or 
       
         
           
           
               
               
           
         
       
     
     
         19 . (canceled) 
     
     
         20 . The moenomycin derivative of  claim 16 , wherein X 1  is hydrogen, 
       
         
           
           
               
               
           
         
       
     
     
         21 - 22 . (canceled) 
     
     
         23 . The moenomycin derivative of  claim 16 , wherein the structure is: 
       
         
           
           
               
               
           
         
         R 3  is selected from the group consisting of hydrogen and hydroxyl, 
         or a pharmaceutically acceptable salt, tautomer, and/or ester thereof. 
       
     
     
         24 . The moenomycin derivative of  claim 23 , wherein R3 is hydrogen or hydroxyl. 
     
     
         25 . (canceled) 
     
     
         26 . The moenomycin derivative of  claim 16 , wherein the structure is: 
       
         
           
           
               
               
           
         
         R 4  is selected from the group consisting of hydrogen and hydroxyl, 
         or a pharmaceutically acceptable salt, tautomer, and/or ester thereof. 
       
     
     
         27 - 28 . (canceled) 
     
     
         29 . The moenomycin derivative of  claim 16 , wherein the structure is: 
       
         
           
           
               
               
           
         
         R 4  is selected from the group consisting of hydrogen and hydroxyl, 
         or a pharmaceutically acceptable salt, tautomer, and/or ester thereof. 
       
     
     
         30 - 31 . (canceled) 
     
     
         32 . The moenomycin derivative of  claim 16 , wherein the structure is: 
       
         
           
           
               
               
           
         
         R 4  is selected from the group consisting of hydrogen and hydroxyl, 
         or a pharmaceutically acceptable salt, tautomer, and/or ester thereof. 
       
     
     
         33 - 34 . (canceled) 
     
     
         35 . The moenomycin derivative of  claim 16 , wherein the structure is: 
       
         
           
           
               
               
           
         
         R 4  is selected from the group consisting of hydrogen and hydroxyl, PG P -1 51 , 
         or a pharmaceutically acceptable salt, tautomer, and/or ester thereof. 
       
     
     
         36 - 37 . (canceled) 
     
     
         38 . The moenomycin derivative of  claim 16 , wherein the structure is: 
       
         
           
           
               
               
           
         
         wherein R 4  is hydrogen or hydroxyl and R 6  is selected from the group consisting of hydrogen, alkyl, cycloalkyl, and substituted cycloalkyl, 
         or a pharmaceutically acceptable salt, tautomer, and/or ester thereof. 
       
     
     
         39 - 43 . (canceled) 
     
     
         44 . A pharmaceutical composition comprising the moenomycin derivative of  claim 16  and a pharmaceutically acceptable carrier. 
     
     
         45 . A moenomycin derivative having the structure: 
       
         
           
           
               
               
           
         
       
       wherein
 R 7  and R 8  independently are selected from the group consisting of hydroxyl and —NHR 9  where R 9  is selected from the group consisting of hydrogen, alkyl, cycloalkyl, and substituted cycloalkyl; and 
 R 10  is hydrogen or hydroxyl; 
 or a pharmaceutically acceptable salt, tautomer, and/or ester thereof. 
 
     
     
         46 - 48 . (canceled) 
     
     
         49 . A pharmaceutical composition comprising the moenomycin derivative of  claim 45  and a pharmaceutically acceptable carrier. 
     
     
         50 . An isolated  Streptomyces  strain selected from the group consisting of:  Streptomyces ghanaensis, Streptomyces ederensis, Streptomyces geysiriensis , and  Streptomyces bambergiensis  strain which carries a one or more mutant or inactivated genes, wherein the mutant or inactivated genes are selected from the group consisting of: moeA4, moeB4, moeC4, moeB5, moe A5, moeD5, moeJ5, moeE5, moeF5, moeH5, moeK5, moeM5, moeN5, moeO5, moeX5, moeP5, moeR5, moeS5, moeGT1, moeGT2, moeGT3, moeGT4, and moeGT5. 
     
     
         51 . The isolated  Streptomyces  strain of  claim 50 , wherein the  Streptomyces ghanaensis  strain is  Streptomyces ghanaensis  ATCC14627. 
     
     
         52 - 54 . (canceled) 
     
     
         55 . The method according to  claim 12 , wherein the method further comprises reacting the moenomycin, moenomycin derivative and/or moenomycin intermediate with a one or more reactants selected from the group consisting of: UDP-sugars, prenyl-pyrophosphates, phosphoglycerate, amino acids, carbamoyl phosphate, ATP and biological cofactors. 
     
     
         56 . The method according to  claim 13 , wherein the method further comprises reacting the moenomycin, moenomycin derivative and/or moenomycin intermediate with a one or more reactants selected from the group consisting of: UDP-sugars, prenyl-pyrophosphates, phosphoglycerate, amino acids, carbamoyl phosphate, ATP and biological cofactors.