Process for the preparation of oxidized phospholipids

Abstract

Novel synthetic routes, which are highly applicable for industrial preparation of therapeutically beneficial oxidized phospholipids are disclosed. Particularly, novel methods for efficiently preparing compounds having a glycerolic backbone and one or more oxidized moieties attached to the glycerolic backbone, which are devoid of column chromatography are disclosed. Further disclosed are novel methods of introducing phosphorous-containing moieties such as phosphate moieties to compounds having glycerolic backbone and intermediates formed thereby.

1 . A method of preparing a compound having a glycerolic backbone and at least one oxidized moiety-containing residue attached to the glycerolic backbone via an ether bond, the method comprising:
 providing a first compound having a glycerolic backbone and at least one free hydroxyl group;   providing a second compound having at least one unsaturated bond and at least one reactive group capable of forming an ether bond with said free hydroxyl group;   reacting said first compound and said second compound to thereby obtain a third compound, said third compound having a glycerolic backbone and an unsaturated bond-containing residue being attached to said glycerolic backbone via an ether bond;   isolating said third compound, to thereby obtain a purified third compound;   reacting said purified third compound with an oxidizing agent, to thereby obtain a fourth compound, said fourth compound having a glycerolic backbone and an oxidized moiety-containing residue attached to said glycerolic backbone via an ether bond; and   isolating said fourth compound to thereby obtain a purified fourth compound, thereby obtaining the compound having a glycerolic backbone and at least one oxidized moiety-containing residue attached to said glycerolic backbone via an ether bond,   the method being devoid of column chromatography.   
     
     
         2 . The method of  claim 1 , wherein isolating said third compound comprises:
 collecting said third compound;   providing a solution of said third compound in a solvent, said solvent being selected such that said third compound is soluble therein whereby impurities formed during said reacting are insoluble therein, to thereby provide a mixture including said solution of said third compound in said solvent and insoluble impurities;   removing said insoluble impurities; and   removing said solvent, thereby obtaining said purified third compound.   
     
     
         3 . The method of  claim 1 , wherein said oxidizing agent is selected from the group consisting of formic acid, hydrogen peroxide, a periodate, a perchlorate, a bismuthate, a permanganate, a chlorite, ozone, silver oxide, osmium tetraoxide and any combination thereof. 
     
     
         4 . The method of  claim 1 , wherein said oxidized moiety is selected from the group consisting of a carboxylic acid, an ester, an aldehyde, an acetal, a ketal and a diol. 
     
     
         5 . The method of  claim 4 , wherein said oxidized moiety is aldehyde and reacting said purified third compound with said oxidizing agent comprises:
 converting said purified third compound to a compound having a glycerolic backbone and a diol-containing residue attached to said glycerolic backbone via an ether bond; and   oxidizing said compound having a glycerolic backbone and a diol-containing residue attached to said glycerolic backbone, to thereby obtain said fourth compound having a glycerolic backbone and an aldehyde-containing residue attached to said glycerolic backbone via an ether bond.   
     
     
         6 . The method of  claim 5 , wherein said converting is effected by reacting said purified third compound with a first oxidizing agent selected from the group consisting of a peroxide, a bismuthate, a periodate, a permanganate, and any combination thereof. 
     
     
         7 . The method of  claim 6 , wherein said oxidizing is effected by reacting said compound having a glycerolic backbone and a diol-containing residue attached to said glycerolic backbone with a second oxidizing agent selected from the group consisting of a periodate, a bismuthate, a permanganate, and a chlorite. 
     
     
         8 . The method of  claim 5 , wherein isolating said fourth compound comprises:
 collecting said fourth compound;   providing a water-soluble adduct of said fourth compound;   subjecting said water-soluble adduct to a biphasic system, to thereby provide an aqueous phase containing said adduct and an organic phase containing water-insoluble impurities formed during said reacting with said oxidizing agent;   collecting said aqueous phase;   decomposing said adduct; and   collecting said fourth compound, thereby obtaining said purified fourth compound.   
     
     
         9 . The method of  claim 4 , wherein said oxidized moiety is a carboxylic acid and reacting said purified third compound with said oxidizing agent comprises:
 converting said purified third compound to a compound having a glycerolic backbone and an aldehyde-containing residue attached to said glycerolic backbone via an ether bond; and   oxidizing said compound having a glycerolic backbone and an aldehyde-containing residue attached to said glycerolic backbone, to thereby obtain a compound having a glycerolic backbone and a carboxylic acid-containing residue attached to said glycerolic backbone via an ether bond.   
     
     
         10 . The method of  claim 4 , wherein said oxidized moiety is a carboxylic acid and reacting said purified third compound with said oxidizing agent comprises:
 converting said purified third compound to a compound having a glycerolic backbone and an epoxide-containing residue attached to said glycerolic backbone via an ether bond; and   oxidizing said compound having a glycerolic backbone and an epoxide-containing residue attached to said glycerolic backbone, to thereby obtain a compound having a glycerolic backbone and a carboxylic acid-containing residue attached to said glycerolic backbone via an ether bond.   
     
     
         11 . The method of  claim 10 , wherein said converting comprises reacting said third compound with a peroxide. 
     
     
         12 . The method of  claim 4 , wherein said oxidized moiety is a carboxylic acid and reacting said purified third compound with said oxidizing agent comprises reacting said purified third compound with a mixture of a permanganate and a periodate. 
     
     
         13 . The method of  claim 1 , wherein said first compound has at least two free hydroxyl groups, the method further comprising, prior to said reacting said first compound and said second compound:
 protecting at least one of said at least two groups with a protecting group.   
     
     
         14 . The method of  claim 10 , wherein said first compound has at least two free hydroxyl groups, the method further comprising, prior to said reacting said first compound and said second compound:
 protecting at least one of said at least two groups with a protecting group.   
     
     
         15 . The method of  claim 1 , wherein the compound having a glycerolic backbone and at least one oxidized moiety-containing residue attached to the glycerolic backbone further comprises a phosphorous-containing moiety attached to the glycerolic backbone, the method further comprising, prior to reacting said first compound and said second compound, prior to isolating said third compound, prior to reacting said third compound with said oxidizing agent, prior to isolating said fourth compound or subsequent to isolating said fourth compound:
 reacting said first compound, said third compound, said purified third compound, said fourth compound or said purified fourth compound with a phosphorous-containing moiety, to thereby obtain said compound having a glycerolic backbone and at least one oxidized moiety-containing residue attached to the glycerolic backbone and further having a phosphorous-containing moiety attached to the glycerolic backbone.   
     
     
         16 . The method of  claim 15 , wherein reacting said first compound, said third compound, said purified third compound, said fourth compound or said purified fourth compound with said phosphorous-containing moiety comprises:
 providing said first compound, said third compound, said purified third compound, said fourth compound or said purified fourth compound having a free hydroxyl group;   reacting said first compound, said third compound, said purified third compound, said fourth compound or said purified fourth compound with a reactive phosphorous-containing compound having a second reactive group and a third reactive group, said second reactive group being capable of reacting with said free hydroxyl group and a second reactive group, to thereby provide said first compound, said third compound, said purified third compound, said fourth compound or said purified fourth compound having a reactive phosphorous-containing group attached to the glycerolic backbone; and   converting said reactive phosphorous-containing group to said phosphorous-containing moiety.   
     
     
         17 . The method of  claim 16 , wherein said reactive phosphorous-containing compound is phosphorous oxychloride (POCl 3 ). 
     
     
         18 . The method of  claim 1 , wherein said first compound has the general formula I: 
       
         
           
           
               
               
           
         
       
       wherein:
 A 1  is absent or is selected from the group consisting of CH 2 , CH═CH and C═O; 
 R 1  is selected from the group consisting of H and a hydrocarbon chain having from 1 to 30 carbon atoms; and 
 R 3  is selected from the group consisting of hydrogen, alkyl, aryl, phosphoric acid, phosphoryl choline, phosphoryl ethanolamine, phosphoryl serine, phosphatidyl choline, phosphatidyl ethanolamine, phosphatidyl serine, phosphatidyl cardiolipin, phosphatidyl inositol, phosphoryl cardiolipin, phosphoryl inositol, ethylphosphocholine, phosphorylmethanol, phosphorylethanol, phosphorylpropanol, phosphorylbutanol, phosphorylethanolamine-N-lactose, phosphoethanolamine-N-[methoxy(propylene glycol)], phosphoinositol-4-phosphate, phosphoinositol-4,5-biposphonate, pyrophosphate, phosphoethanolamine-diethylenetriamine-pentaacetate, dinitrophenyl-phosphoethanolamine, phosphoglycerol. 
 
     
     
         19 . The method of  claim 18 , wherein the compound having a glycerolic compound and at least one oxidized moiety attached to the glycerolic backbone via an ether bond has the general Formula II: 
       
         
           
           
               
               
           
         
       
       wherein:
 A 1  is selected from the group consisting of CH 2 , CH═CH and C═O; 
 A 2  is CH 2 ; 
 R 1  is an alkyl having 1-30 carbon atoms; 
 R 2  is 
 
       
         
           
           
               
               
           
         
       
       whereas:
 X is an alkyl chain having 1-24 carbon atoms; 
 Y is selected from the group consisting of hydrogen, hydroxy, alkyl, alkoxy, halide, acetoxy and an aromatic functional group; and 
 Z is selected from the group consisting of: 
 
       
         
           
           
               
               
           
         
         with R 4  being an alkyl or aryl; and 
         R 3  is selected from the group consisting of hydrogen, alkyl, aryl, phosphoric acid, phosphoryl choline, phosphoryl ethanolamine, phosphoryl serine, phosphatidyl choline, phosphatidyl ethanolamine, phosphatidyl serine, phosphatidyl cardiolipin, phosphatidyl inositol, phosphoryl cardiolipin, phosphoryl inositol, ethylphosphocholine, phosphorylmethanol, phosphorylethanol, phosphorylpropanol, phosphorylbutanol, phosphorylethanolamine-N-lactose, phosphoethanolamine-N-[methoxy(propylene glycol)], phosphoinositol-4-phosphate, phosphoinositol-4,5-biposphonate, pyrophosphate, phosphoethanolamine-diethylenetriamine-pentaacetate, dinitrophenyl-phosphoethanolamine, phosphoglycerol. 
       
     
     
         20 . The method of  claim 19 , wherein A 1  is CH 2 .