Lipidated psa compositions and methods

Abstract

The invention provides various isolated and synthetic forms of lipidated PSA as well as isolated or synthetic forms of the glycolipid component of lipidated PSA, and compositions thereof, methods of making including methods of isolating such forms, and methods of use thereof.

1 . An isolated lipidated polysaccharide A (PSA) comprising
 polysaccharide A (PSA) covalently conjugated to a glycolipid, wherein the glycolipid is di-acylated, tri-acylated, tetra-acylated or penta-acylated.   
     
     
         2 - 40 . (canceled) 
     
     
         41 . An isolated glycolipid comprising a diglucosamine covalently conjugated to 2-5 acyl chains, each independently ranging in length from 14-17 carbons. 
     
     
         42 . The isolated glycolipid of  claim 41 , wherein the diglucosamine is covalently conjugated to 2-4 acyl chains. 
     
     
         43 . The isolated glycolipid of  claim 41 , wherein the diglucosamine is covalently conjugated to 4 acyl chains. 
     
     
         44 . The isolated glycolipid of  claim 41 , wherein the acyl chains range in length from 15-17 carbons. 
     
     
         45 . A non-hydrolytic method for isolating lipidated polysaccharide A (PSA) from  B. fragilis , comprising
 extracting, into an aqueous phase, capsular complex from  B. fragilis  using a mixture of phenol and water,   precipitating a polysaccharide fraction from the aqueous phase using ethanol, and   isolating lipidated PSA from the polysaccharide fraction by size exclusion.   
     
     
         46 - 100 . (canceled) 
     
     
         101 . A method comprising
 administering, to a subject having or at risk of developing a condition associated with inflammation, an effective amount of the isolated glycolipid of  claim 41 .   
     
     
         102 - 110 . (canceled) 
     
     
         111 . The isolated glycolipid of  claim 41 , wherein the diglucosamine is covalently conjugated to 5 acyl chains. 
     
     
         112 . The isolated glycolipid of  claim 41 , wherein at least one of the one or more acyl chains is unmodified and/or at least one of the one or more acyl chains is modified 
     
     
         113 . The isolated glycolipid of  claim 41 , wherein at least one of the one or more acyl chains is modified with a hydroxyl group. 
     
     
         114 . The isolated glycolipid of  claim 41 , wherein at least one of the one or more acyl chains is C16:0-OH, C17:0-OH, C14:0, and/or C15:0. 
     
     
         115 . The isolated glycolipid of  claim 41 , wherein at least one of the one or more acyl chains is N-substituted on a disaccharide and/or at least one of the one or more acyl chains is O-substituted on a disaccharide. 
     
     
         116 . The isolated glycolipid of  claim 41 , substantially free of other components found in a  B. fragilis  capsule, substantially free of LPS, substantially free of unconjugated glycolipid, and/or free of non-lipidated PSA. 
     
     
         117 . The isolated glycolipid of  claim 41 , in purified form, in a micelle form, and/or in lyophilized form. 
     
     
         118 . The isolated glycolipid of  claim 41 , in a pharmaceutical composition, optionally wherein the pharmaceutical composition is suitable for administration to a human, optionally wherein the pharmaceutical composition comprises less than 0.5% (w/w) of free glycolipid, optionally wherein the pharmaceutical composition comprises a detergent or a bile salt.