PROCESS FOR PRODUCING HMG-CoA REDUCTASE INHIBITORS

Abstract

The present invention relates to processes for producing compounds by using a protein derived from a microorganism belonging to the genus Bacillus , which has an activity of hydroxylating a compound represented by the formula (I-a): wherein R 1 represents a hydrogen atom, a substituted or unsubstituted alkyl, or an alkali metal, and R 2 represents a substituted or unsubstituted alkyl or substituted or unsubstituted aryl, or a ring-closed lactone from thereof.

1 . A process for producing compound (II-a) or compound (II-b), wherein an isolated transformed cell obtained by introducing into a host cell a recombinant DNA vector comprising the DNA having the nucleotide sequence shown by SEQ ID NO: 2, a culture of the transformed cell, or a treated product of the culture is used as an enzyme source, and the process comprises:
 allowing compound (I-a) or compound (I-b) to exist in an aqueous medium in the presence of the enzyme source;   allowing compound (II-a) or compound (II-b) to be produced and accumulated in said aqueous medium; and   collecting compound (II-a) or compound (II-b) from said aqueous medium wherein, the compound (I-a) is a compound represented by the formula (I-a):   
       
         
           
           
               
               
           
         
       
       the compound (I-b) is a lactone form of compound (I-a) and is represented by the formula (I-b): 
       
         
           
           
               
               
           
         
       
       the compound (II-a) is a compound represented by the formula (II-a): 
       
         
           
           
               
               
           
         
       
       the compound (II-b) is a lactone form of compound (II-a) and is represented by the formula (II-b): 
       
         
           
           
               
               
           
         
       
       and wherein R 1  represents a hydrogen atom, a substituted or unsubstituted alkyl, or an alkali metal, and R 2  represents a substituted or unsubstituted alkyl, or a substituted or unsubstituted aryl; and wherein the treated product of the culture of the transformed cell is a treated product selected from cultured cells, dried cells, freeze-dried cells, cells treated with a surfactant, cells treated with an enzyme, cells treated by ultrasonication, cells treated by mechanical milling, cells treated by solvent, a protein fraction of a cell, and immobilized cells. 
     
     
         2 . A process for producing compound (IV-a) or compound (IV-b), wherein an isolated transformed cell obtained by introducing into a host cell a recombinant DNA vector comprising the DNA having the nucleotide sequence shown by SEQ ID NO: 2, a culture of the transformed cell, or a treated product of the culture is used as an enzyme source, and the process comprises:
 allowing compound (III-a) or compound (III-b) to exist in an aqueous medium in the presence of the enzyme source;   allowing compound (IV-a) or compound (IV-b) to be produced and accumulated in said aqueous medium; and   collecting compound (IV-a) or compound (IV-b) from said aqueous medium wherein, the compound (III-a) is a compound represented by the formula (III-a):   
       
         
           
           
               
               
           
         
       
       the compound (III-b) is a lactone form of compound (III-a) and is represented by the formula (III-b): 
       
         
           
           
               
               
           
         
       
       the compound (IV-a) is a compound represented by the formula (IV-a): 
       
         
           
           
               
               
           
         
       
       the compound (IV-b) is a lactone form of compound (IV-a) and is represented by the formula (IV-b): 
       
         
           
           
               
               
           
         
       
       and wherein R 1  represents a hydrogen atom, a substituted or unsubstituted alkyl, or an alkali metal, and R 2  represents a substituted or unsubstituted alkyl, or a substituted or unsubstituted aryl; and wherein the treated product of the culture of the transformed cell is a treated product selected from cultured cells, dried cells, freeze-dried cells, cells treated with a surfactant, cells treated with an enzyme, cells treated by ultrasonication, cells treated by mechanical milling, cells treated by solvent, a protein fraction of a cell, and immobilized cells. 
     
     
         3 . A process for producing compound (VI-a) or compound (VI-b), wherein an isolated transformed cell obtained by introducing into a host cell a recombinant DNA vector comprising the DNA having the nucleotide sequence shown by SEQ ID NO: 2, a culture of the transformed cell, or a treated product of the culture is used as an enzyme source, and the process comprises:
 allowing compound (V-a) or compound (V-b) to exist in an aqueous medium in the presence of the enzyme source;   allowing compound (VI-a) or compound (VI-b) to be produced and accumulated in said aqueous medium; and   collecting compound (VI-a) or compound (VI-b) from said aqueous medium wherein, the compound (V-a) is a compound represented by the formula (V-a):   
       
         
           
           
               
               
           
         
       
       the compound (V-b) is a lactone form of compound (V-a) and is represented by the formula (V-b): 
       
         
           
           
               
               
           
         
       
       the compound (VI-a) is a compound represented by the formula (VI-a): 
       
         
           
           
               
               
           
         
       
       the compound (VI-b) is a lactone form of compound (VI-a) and is represented by the formula (VI-b): 
       
         
           
           
               
               
           
         
       
       and wherein R 1  represents a hydrogen atom, a substituted or unsubstituted alkyl, or an alkali metal; and wherein the treated product of the culture of the transformed cell is a treated product selected from cultured cells, dried cells, freeze-dried cells, cells treated with a surfactant, cells treated with an enzyme, cells treated by ultrasonication, cells treated by mechanical milling, cells treated by solvent, a protein fraction of a cell, and immobilized cells. 
     
     
         4 . A process for producing compound (VIII-a) or compound (VIII-b), wherein an isolated transformed cell obtained by introducing into a host cell a recombinant DNA vector comprising the DNA having the nucleotide sequence shown by SEQ ID NO: 2, a culture of the transformed cell, or a treated product of the culture is used as an enzyme source, and the process comprises:
 allowing compound (VII-a) or compound (VII-b) to exist in an aqueous medium in the presence of the enzyme source;   allowing compound (VII-a) or compound (VII-b) to be produced and accumulated in said aqueous medium; and   collecting compound (VIII-a) or compound (VIII-b) from said aqueous medium wherein, the compound (VII-a) is a compound represented by the formula (VII-a):   
       
         
           
           
               
               
           
         
       
       the compound (VII-b) is a lactone form of compound (VII-a) and is represented by the formula (VII-b): 
       
         
           
           
               
               
           
         
       
       the compound (VIII-a) is a compound represented by the formula (VIII-a): 
       
         
           
           
               
               
           
         
       
       the compound (VIII-b) is a lactone form of compound (VIII-a) and is represented by the formula (VIII-b): 
       
         
           
           
               
               
           
         
       
       and wherein R 1  represents a hydrogen atom, a substituted or unsubstituted alkyl, or an alkali metal; and wherein the treated product of the culture of the transformed cell is a treated product selected from cultured cells, dried cells, freeze-dried cells, cells treated with a surfactant, cells treated with an enzyme, cells treated by ultrasonication, cells treated by mechanical milling, cells treated by solvent, a protein fraction of a cell, and immobilized cells. 
     
     
         5 . The process according to  claim 1 , wherein the compound (II-b) is obtained by forming a lactone from compound (II-a), the compound (II-a) is a compound represented by the formula (II-a): 
       
         
           
           
               
               
           
         
       
       the compound (II-b) is a lactone form of compound (II-a) and is represented by the formula (II-b): 
       
         
           
           
               
               
           
         
       
       and wherein R 1  represents a hydrogen atom, a substituted or unsubstituted alkyl, or an alkali metal, and R 2  represents a substituted or unsubstituted alkyl, or a substituted or unsubstituted aryl. 
     
     
         6 . The process according to  claim 1 , wherein the compound (II-a) is obtained by opening the lactone ring of compound (II-b), the compound (II-a) is a compound represented by the formula (II-a): 
       
         
           
           
               
               
           
         
       
       the compound (II-b) is a lactone form of compound (II-a) and is represented by the formula (II-b): 
       
         
           
           
               
               
           
         
       
       and wherein R 1  represents a hydrogen atom, a substituted or unsubstituted alkyl, or an alkali metal, and R 2  represents a substituted or unsubstituted alkyl, or a substituted or unsubstituted aryl. 
     
     
         7 . The process according to  claim 2 , wherein the compound (IV-b) is obtained by forming a lactone from compound (IV-a), the compound (IV-a) is a compound represented by the formula (IV-a): 
       
         
           
           
               
               
           
         
       
       the compound (IV-b) is a lactone form of compound (IV-a) and is represented by the formula (IV-b): 
       
         
           
           
               
               
           
         
       
       and wherein R 1  represents a hydrogen atom, a substituted or unsubstituted alkyl, or an alkali metal, and R 2  represents a substituted or unsubstituted alkyl, or a substituted or unsubstituted aryl. 
     
     
         8 . The process according to  claim 2 , wherein the compound (IV-a) is obtained by opening the lactone ring of compound (IV-b), the compound (IV-a) is a compound represented by the formula (IV-a): 
       
         
           
           
               
               
           
         
       
       the compound (IV-b) is a lactone form of compound (IV-a) and is represented by the formula (IV-b): 
       
         
           
           
               
               
           
         
       
       and wherein R 1  represents a hydrogen atom, a substituted or unsubstituted alkyl, or an alkali metal, and R 2  represents a substituted or unsubstituted alkyl, or a substituted or unsubstituted aryl. 
     
     
         9 . The process according to  claim 3 , wherein the compound (VI-b) is obtained by forming a lactone from compound (VI-a), the compound (VI-a) is a compound represented by the formula (VI-a): 
       
         
           
           
               
               
           
         
       
       the compound (VI-b) is a lactone form of compound (VI-a) and is represented by the formula (VI-b): 
       
         
           
           
               
               
           
         
       
       and wherein R 1  represents a hydrogen atom, a substituted or unsubstituted alkyl, or an alkali metal. 
     
     
         10 . The process according to  claim 3 , wherein the compound (VI-a) is obtained by opening the lactone ring of compound (VI-b), the compound (VI-a) is a compound represented by the formula (VI-a): 
       
         
           
           
               
               
           
         
       
       the compound (VI-b) is a lactone form of compound (VI-a) and is represented by the formula (VI-b): 
       
         
           
           
               
               
           
         
       
       and wherein R 1  represents a hydrogen atom, a substituted or unsubstituted alkyl, or an alkali metal. 
     
     
         11 . The process according to  claim 4 , wherein the compound (VIII-b) is obtained by forming a lactone from compound (VIII-a), the compound (VIII-a) is a compound represented by the formula (VIII-a): 
       
         
           
           
               
               
           
         
       
       the compound (VIII-b) is a lactone form of compound (VIII-a) and is represented by the formula (VIII-b): 
       
         
           
           
               
               
           
         
       
       and wherein R 1  represents a hydrogen atom, a substituted or unsubstituted alkyl, or an alkali metal. 
     
     
         12 . The process according to  claim 4 , wherein the compound (VIII-a) is obtained by opening the lactone ring of compound (VIII-b), the compound (VIII-a) is a compound represented by the formula (VIII-a): 
       
         
           
           
               
               
           
         
       
       the compound (VIII-b) is a lactone form of compound (VIII-a) and is represented by the formula (VIII-b): 
       
         
           
           
               
               
           
         
       
       and wherein R 1  represents a hydrogen atom, a substituted or unsubstituted alkyl, or an alkali metal. 
     
     
         13 . A process for producing a protein, which is derived from a microorganism belonging to the genus  Bacillus , and has an activity of producing compound (II-a) or compound (II-b) from compound (I-a) or compound (I-b). 
       wherein the compound (I-a) is a compound represented by the formula (I-a): 
       
         
           
           
               
               
           
         
       
       the compound (I-b) is a lactone form of compound (I-a) and is represented by the formula (I-b): 
       
         
           
           
               
               
           
         
       
       the compound (II-a) is a compound represented by the formula (II-a): 
       
         
           
           
               
               
           
         
       
       the compound (II-b) is a lactone form of compound (II-a) and is represented by the formula (II-b): 
       
         
           
           
               
               
           
         
       
       wherein R 1  represents a hydrogen atom, a substituted or unsubstituted alkyl, or an alkali metal, and R 2  represents a substituted or unsubstituted alkyl, or a substituted or unsubstituted aryl, which comprises culturing a transformed cell obtained by introducing a recombinant DNA vector comprising the DNA having the nucleotide sequence shown by SEQ ID NO: 2 in a medium; producing and accumulating the protein in the culture; and collecting the protein from said culture.