Polysaccharide compositions and methods of use for the treatment and prevention of disorders associated with progenitor cell mobilization

Abstract

Polysaccharide preparations lacking substantial anticoagulant activity are provided herein. Methods of making and using such preparations are provided.

1 - 34 . (canceled) 
     
     
         35 . A pharmaceutical composition comprising a low molecular weight heparin preparation produced by a process comprising:
 depolymerization of unfractionated heparin to produce a depolymerized polysaccharide preparation,   oxidation of the depolymerized polysaccharide preparation to produce an oxidized polysaccharide preparation, and   reduction of the oxidized polysaccharide preparation to produce the low molecular weight heparin preparation, wherein the low molecular weight heparin preparation has the following characteristics   (a) a weight average chain molecular weight between 5,000 and 8,000 Da as determined by high performance liquid chromatography coupled with a multi angle light scattering detector (MALS);   (b) anti-Xa activity of less than 20 IU/mg and anti-IIa activity of 1 IU/mg or less;   (c) greater than 5% and less than 25% glycol split uronic acid residues;   (d) a molecular weight distribution such that 10-30% of the oligosaccharides have a molecular weight <3000 Da; 55-65% of the oligosaccharides have a molecular weight between 3000-8000 Da, and 15-30% of the oligosaccharides have a molecular weight >8000 Da as determined by high performance liquid chromatography coupled with a multi angle light scattering detector (MALS); and   wherein the preparation comprises polysaccharides comprising the structure:   [U w —H x,y,z ] m ˜[U G —H x,y,z ] n , or a pharmaceutically acceptable salt thereof,   wherein each occurrence of U indicates a uronic acid residue and each occurrence of H indicates a hexosamine residue;   wherein m and n are integers such that   m=4-15, and   n=1-3;   each of w, x, y and z can, independently, be the same or different for each occurrence of [U w —H x,y,z ] and each of x, y and z can, independently, be the same or different for each occurrence of [U G —H x,y,z ], wherein   w=−2OS or −2OH;   x=−NS or −NAc;   y=−3OS or −3OH;   z=−6OS or −6OH;   and   
       
         
           
           
               
               
           
         
         wherein the symbol ˜ indicates that the units marked m and n are distributed along the polysaccharide chain and are not necessarily in sequence. 
       
     
     
         36 . The pharmaceutical composition of  claim 35 , wherein the preparation has greater than 5% and less than 20% glycol split uronic acid residues. 
     
     
         37 . The preparation of  claim 35 , wherein the pharmaceutical composition has between 10% and 20% glycol split uronic acid residues. 
     
     
         38 . The pharmaceutical composition of  claim 35 , wherein the polysaccharide preparation has a plurality of chains having a glycol split uronic acid residue. 
     
     
         39 . The pharmaceutical composition of  claim 38 , wherein each polysaccharide chain of the preparation has no more than 2 glycol split uronic acid residues (U G ). 
     
     
         40 . The pharmaceutical composition of  claim 35 , wherein the polysaccharide preparation has greater than 40% U 2S H NS,6S  disaccharide residues. 
     
     
         41 . The pharmaceutical composition of  claim 40 , wherein the polysaccharide preparation has greater than 70% U 2S H NS,6S  disaccharide residues. 
     
     
         42 . The pharmaceutical composition of  claim 40 , wherein the polysaccharide preparation has a degree of desulfation less than 40%. 
     
     
         43 . The pharmaceutical composition of  claim 40 , wherein the polysaccharide preparation has a degree of desulfation less than 30%. 
     
     
         44 . The pharmaceutical composition of  claim 40 , wherein the polysaccharide preparation has a degree of desulfation less than 10%. 
     
     
         45 . The pharmaceutical composition of  claim 35 , wherein the reducing end of the polysaccharides comprise a 2,5-anhydromannitol residue. 
     
     
         46 . The pharmaceutical composition of  claim 35 , wherein about 50% of the reducing ends of the polysaccharides comprise a 2,5-anhydromannitol residue. 
     
     
         47 . The pharmaceutical composition of  claim 35 , wherein the polysaccharides of the preparation have a uronic acid at the non-reducing end. 
     
     
         48 . The pharmaceutical composition of  claim 47 , wherein the polysaccharides of the preparation have a non native uronic acid at the non-reducing end. 
     
     
         49 . The pharmaceutical composition of  claim 47 , wherein the polysaccharides of the preparation have a glycol split uronic acid at the non-reducing end. 
     
     
         50 . The pharmaceutical composition of  claim 35 , wherein the weight average chain molecular weight of the preparation is between 4,000 and 8,000 Da. 
     
     
         51 . The pharmaceutical composition of  claim 50 , wherein the reducing end of the polysaccharides comprise a 2,5-anhydromannitol residue. 
     
     
         52 . The pharmaceutical composition of  claim 50 , wherein about 50% of the reducing ends of the polysaccharides comprise a 2,5-anhydromannitol residue. 
     
     
         53 . The pharmaceutical composition of  claim 50 , wherein the polysaccharides of the preparation have a uronic acid at the non-reducing end. 
     
     
         54 . The pharmaceutical composition of  claim 53 , wherein the polysaccharides of the preparation have a non native uronic acid at the non-reducing end. 
     
     
         55 . The pharmaceutical composition of  claim 53 , wherein the polysaccharides of the preparation have a glycol split uronic acid at the non-reducing end. 
     
     
         56 . The pharmaceutical composition of  claim 35 , wherein the polysaccharide preparation has an anti-Xa activity of less than 15 IU/mg. 
     
     
         57 . The pharmaceutical composition of  claim 35 , wherein the polysaccharide preparation has an anti-Xa activity of less than 10 IU/mg. 
     
     
         58 . The pharmaceutical composition of  claim 35 , wherein the preparation has a polydispersity of about 1.2 to 1.7. 
     
     
         59 . The pharmaceutical composition of  claim 35 , wherein the composition has one or more of a sodium content less than 30%; less than 20 ppm iodine; less than 30% sulfur; and/or less than 50 ppm boron. 
     
     
         60 . The pharmaceutical composition of  claim 35  further comprising a pharmaceutically acceptable carrier. 
     
     
         61 . The pharmaceutical composition of  claim 35  formulated for subcutaneous administration. 
     
     
         62 . A unit dosage form comprising the pharmaceutical composition of  claim 35 . 
     
     
         63 . The unit dosage form of  claim 62 , wherein the unit dosage form is an ampoule, syringe pen, or syringe. 
     
     
         64 . A pharmaceutical composition comprising a low molecular weight heparin preparation produced by a process comprising:
 depolymerization of unfractionated heparin to produce a depolymerized polysaccharide preparation,   oxidation of the depolymerized polysaccharide preparation to produce an oxidized polysaccharide preparation, and   reduction of the oxidized polysaccharide preparation to produce the low molecular weight heparin preparation, wherein the low molecular weight heparin preparation has the following characteristics   (a) a weight average chain molecular weight between 5,000 and 8,000 Da as determined by high performance liquid chromatography coupled with a multi angle light scattering detector (MALS);   (b) anti-Xa activity of less than 20 IU/mg and anti-IIa activity of 1 IU/mg or less;   (c) greater than 5% and less than 25% glycol split uronic acid residues;   (d) a molecular weight distribution such that 10-30% of the oligosaccharides have a molecular weight <3000 Da; 55-65% of the oligosaccharides have a molecular weight between 3000-8000 Da, and 15-30% of the oligosaccharides have a molecular weight >8000 Da as determined by high performance liquid chromatography coupled with a multi angle light scattering detector (MALS); and   wherein the preparation comprises polysaccharides comprising the structure:   [U w —H x,y,z ] m ˜[U G —H x,y,z ] n , or a pharmaceutically acceptable salt thereof,   wherein each occurrence of U indicates a uronic acid residue and each occurrence of H indicates a hexosamine residue;   wherein m and n are integers such that   m=4-15, and   n=1-3;   each of w, x, y and z can, independently, be the same or different for each occurrence of [U w —H x,y,z ] and each of x, y and z can, independently, be the same or different for each occurrence of [U G —H x,y,z ], wherein   w=−2OS or −2OH;   x=−NS or −NAc;   y=−3OS or −3OH;   z=−6OS or −6OH;   and   
       
         
           
           
               
               
           
         
         wherein the symbol ˜ indicates that the units marked m and n are distributed along the polysaccharide chain and are not necessarily in sequence; and 
         wherein a plurality of the polysaccharides in the preparation have a 2,5-anhydromannitol residue at the reducing end and a non-native uronic acid at the non-reducing end. 
       
     
     
         65 . The pharmaceutical composition of  claim 64  further comprising a pharmaceutically acceptable carrier. 
     
     
         66 . The pharmaceutical composition of  claim 64  formulated for subcutaneous administration. 
     
     
         67 . A unit dosage form comprising the pharmaceutical composition of  claim 64 . 
     
     
         68 . The unit dosage form of  claim 67 , wherein the unit dosage form is an ampoule, syringe pen, or syringe.