Compositions and methods for making noscapine and synthesis intermediates thereof

Abstract

Methods that may be used for the manufacture of the therapeutic chemical compound noscapine and noscapine synthesis intermediates. Compounds useful for the synthesis of noscapine are also provided.

1 . A method of making noscapine or a noscapine synthesis intermediate comprising:
 (a) providing a noscapine pathway precursor selected from a first canadine derivative, a first papaveroxine derivative and narcotine hemiacetal; and   (b) contacting the noscapine pathway precursor with at least one of the enzymes selected from the group of enzymes consisting of (i) CYP82Y1; (ii) CYP82X2; (iii) AT1; (iv) CYP82X1; (v) OMT; (vi) CXE1; and (vii) NOS under reaction conditions permitting the catalysis of the noscapine pathway precursor to form noscapine or the noscapine synthesis intermediate wherein the noscapine intermediate is a second canadine derivative, a first or second papaveroxine derivative, narcotine hemiacetal or noscapine;   and   wherein the first canadine derivative has the chemical formula (I):   
       
         
           
           
               
               
           
         
         wherein R 1  represents a hydrogen atom, hydroxyl; or O-acetyl; 
         wherein R 2  represents a hydrogen atom or hydroxyl; and 
         wherein R 3  represents a hydrogen atom or hydroxyl; 
         wherein the second canadine derivative has the chemical formula (II): 
       
       
         
           
           
               
               
           
         
         wherein R 4  represents a hydrogen atom, hydroxyl; or O-acetyl; and 
         wherein R 5  represents a hydrogen atom or hydroxyl; and 
         wherein the first and second papaveroxine derivative have the chemical formula (III): 
       
       
         
           
           
               
               
           
         
         wherein R 6  represents hydroxyl or methoxy; and 
         wherein R 7  represents hydroxyl or O-acetyl. 
       
     
     
         2 . The method according to  claim 1  wherein the first canadine derivative is selected from the group of canadine derivatives consisting of (S)—N-methylcanadine; 1-hydroxy-N-methylcanadine; 1,13-dihydroxy-N-methylcanadine, 1-hydroxy-13-O-acetyl-N-methylcanadine; and 1,8-dihydroxy-13-O-acetyl-N-methyl canadine; and wherein the second canadine derivative is selected from the group of canadine derivatives consisting of 1-hydroxy-N-methylcanadine; 1,13-dihydroxy-N-methylcanadine, 1-hydroxy-13-O-acetyl-N-methylcanadine; and 1,8-dihydroxy-13-O-acetyl-N-methyl canadine; and wherein the first and second papaveroxine derivatives are selected from the group of papaveroxine derivatives consisting of 4′-O-desmethoxy-3-O-acetyl-papaveroxine; papaveroxine; or 3-O-acetyl-papaveroxine. 
     
     
         3 . A method of making noscapine comprising:
 (a) providing a noscapine pathway precursor selected from a canadine derivative, a papaveroxine derivative or narcotine hemiacetal; and   (b) contacting the noscapine pathway precursor with at least one of the enzymes selected from the group of enzymes consisting of (i) CYP82Y1; (ii) CYP82X2; (iii) AT1; (iv) CYP82X1; (v) OMT; (vi) CXE1; and (vii) NOS under reaction conditions permitting the catalysis of the noscapine pathway precursor to form noscapine;   and wherein the canadine derivative has the chemical formula (I):   
       
         
           
           
               
               
           
         
         wherein R 1  represents a hydrogen atom, hydroxyl; or O-acetyl; 
         wherein R 2  represents a hydrogen atom or hydroxyl; and 
         wherein R 3  represents a hydrogen atom or hydroxyl; 
         and wherein the papaveroxine derivative has the chemical formula (III): 
       
       
         
           
           
               
               
           
         
         wherein R 6  represents hydroxyl or methoxy; and 
         wherein R 7  represents hydroxyl or O-acetyl. 
       
     
     
         4 . A method of making noscapine according to  claim 3  wherein the noscapine pathway precursor is (S)—N-methylcanadine; and the enzymes are (i) CYP82Y1; (ii) CYP82X2; (iii) AT1; (iv) CYP82X1; (v) OMT; (vi) CXE1; and (vii) NOS. 
     
     
         5 . A method of making noscapine according to  claim 3  wherein the noscapine pathway precursor is 1-hydroxy-N-methylcanadine; and the enzymes are (i) CYP82X2; (ii) AT1; (iii) CYP82X1; (iv) OMT; (v) CXE1; and (vi) NOS. 
     
     
         6 . A method of making noscapine according to  claim 3  wherein the noscapine pathway precursor is 1,13-dihydroxy-N-methylcanadine; and the enzymes are (i) AT1; (ii) CYP82X1; (iii) OMT; (iv) CXE1; and (v) NOS. 
     
     
         7 . A method of making noscapine according to  claim 3  wherein the noscapine pathway precursor is 1-hydroxy-13-O-acetyl-N-methylcanadine; and the enzymes are (i) CYP82X1; (ii) OMT; (iii) CXE1; and (iv) NOS. 
     
     
         8 . A method of making noscapine according to  claim 3  wherein the noscapine pathway precursor is 1,8-dihydroxy-13-O-acetyl-N-methylcanadine; and the enzymes are (i) OMT; (ii) CXE1; and (iii) NOS. 
     
     
         9 . A method of making noscapine according to  claim 3  wherein the noscapine pathway precursor is 4′-O-desmethoxy-3-O-acetyl-N-papaveroxine; and the enzymes are (i) OMT; (ii) CXE1; and (iii) NOS. 
     
     
         10 . A method of making noscapine according to  claim 3  wherein the noscapine precursor is 3-O-acetyl-N-papaveroxine; and the enzymes are (i) CXE1; and (ii) NOS. 
     
     
         11 . A method of making noscapine according to  claim 3  wherein the noscapine pathway precursor is papaveroxine; and the enzyme is NOS. 
     
     
         12 . A method of making noscapine according to  claim 3  wherein the noscapine pathway precursor is narcotine hemiacetal; and the enzyme is NOS. 
     
     
         13 . The method according to  claim 1  wherein the reaction conditions are in vitro reaction conditions. 
     
     
         14 . The method according to  claim 1  wherein the reaction conditions are in vivo reaction conditions. 
     
     
         15 . A method for preparing noscapine and/or one or more of the noscapine synthesis intermediates selected from the group of noscapine synthesis intermediates consisting of: 1-hydroxy-N-methylcanadine; 1,13-dihydroxy-N-methylcanadine; 1-hydroxy-O-acetyl-N-methylcanadine; 1,8-dihydroxy-13-O-acteyl-N-methylcanadine; 4′-O-desmethoxy-3-O-acetyl-papaveroxine; 3-O-acetyl-papaveroxine; papaveroxine; and narcotine hemiacetal comprising:
 (a) providing a chimeric nucleic acid sequence comprising in the 5′ to 3′ direction of transcription as operably linked components:
 (i) one or more nucleic acid sequences encoding one or more of the polypeptides selected from the group of polypeptides consisting of CYP82Y1; CYP82X2; AT1; CYP82X1; OMT; CXE1 and NOS; and 
 (ii) one or more nucleic acid sequences capable of controlling expression in a host cell; 
 
 (b) introducing the chimeric nucleic acid sequence into a host cell and growing the host cell to produce the polypeptide selected from the group of polypeptides consisting of CYP82Y1; CYP82X2; AT1; CYP82X1; OMT; CXE1 and/or NOS and to produce one or more of noscapine or the noscapine synthesis intermediate; and 
 (c) recovering noscapine or the noscapine synthesis intermediate. 
 
     
     
         16 . The method according to  claim 15  wherein the method is a method for preparing noscapine, and the enzymes are (i) CYP82X1; (ii) OMT; (iii) CXE1; and (iv) NOS. 
     
     
         17 . The method according to  claim 15  wherein the method is a method for preparing noscapine, and the enzymes are (i) OMT; (ii) CXE1; and (iii) NOS. 
     
     
         18 . The method according to  claim 15  wherein the method is a method for preparing noscapine, and the enzymes are (i) OMT; (ii) CXE1; and (iii) NOS. 
     
     
         19 . The method according to  claim 15  wherein the method is a method for preparing noscapine, and the enzymes are (i) CXE1; and (ii) NOS. 
     
     
         20 . The method according to  claim 15  wherein the method is a method for preparing noscapine; and the enzyme is NOS.