Fusogenic properties of saposin C and related proteins and polypeptides for application to transmembrane drug delivery systems

Abstract

The present invention comprises a method for delivering pharmaceutical agents within and/or through the dermal and mucosal membranes, utilizing a fusogenic protein. The fusogenic protein is associated with a phospholipid membrane, such as a liposome. The liposome contains the pharmaceutical agent. Preferred fusogenic proteins include saposin C and other proteins, polypeptides and peptide analogs derived from saposin C. The active agent contained within the liposome may comprise large biomolecules and/or small organic molecules. This technology can be used for both cosmetic and medicinal applications in which the objective is delivery of the active agent within and/or beneath the biological membrane.

1 ) A method for delivering a pharmaceutical agent through a membrane, wherein the method comprises applying to said membrane a composition comprising: 
 a) anionic phospholipids;    b) a safe and effective amount of the pharmaceutical agent contained within the aqueous interior of the phospholipids; and    c) a fusogenic protein or polypeptide derived from prosaposin    in a pharmaceutically acceptable carrier, wherein the concentration of the fusogenic protein or polypeptide is of a sufficient amount to deliver the pharmaceutical agent through the membrane.    
     
     
         2 ) The method of  claim 2  wherein the concentration of phospholipids are in at least a 10-fold excess, by weight, to that of the fusogenic protein or polypeptide.  
     
     
         3 ) The method of  claim 2  wherein the pH of the composition is between about 5.5 and 2.  
     
     
         4 ) The method of  claim 3  wherein the anionic phospholipid is an anionic liposome.  
     
     
         5 ) The method of  claim 4  wherein the fusogenic protein or polypeptide is associated with the liposome through an electrostatic and hydrophobic interaction.  
     
     
         6 ) The method of  claim 5  wherein the membrane is selected from the group consisting of dermal and mucosal membranes.  
     
     
         7 ) The method of  claim 6  wherein the fusogenic protein or polypeptide is selected from the group consisting of saposin A, saposin C, and mixtures thereof.  
     
     
         8 ) The method of  claim 6  wherein the fusogenic protein or polypeptide is saposin C.  
     
     
         9 ) The method of  claim 6  wherein the fusogenic protein or polypeptide is SEQ. ID. NO. 1.  
     
     
         10 ) The method of  claim 6  wherein the fusogenic protein or polypeptide is SEQ. ID. NO. 2.  
     
     
         11 . The method of  claim 6  wherein the fusogenic protein or polypeptide is selected from the group consisting of those proteins or polypeptides given by SEQ ID Nos. 3-6.  
     
     
         12 ) The method of  claim 7  wherein administration of the composition is via a transdermal patch.  
     
     
         13 ) The method of  claim 7  wherein the composition is administered either enterally or topically.  
     
     
         14 ) A method for delivering a pharmaceutical agent through either a dermal or mucosal membrane, wherein the method comprises the administration to said membrane of a composition comprising: 
 a) anionic liposomes;    b) a safe and effective amount of the pharmaceutical agent contained within the aqueous interior of the liposomes; and    c) saposin C;    in a pharmaceutically acceptable carrier, wherein the concentration of the liposomes are of a sufficient amount to deliver a safe and effective amount of the pharmaceutical agent through the membrane, the pH of the composition is between about 5.5 and 2, and the saposin C is associated with the surface of the liposome through an electrostatic and hydrophobic interaction.    
     
     
         15 ) The method of  claim 14  wherein the concentration of the liposomes is in at least a 10-fold excess, by weight, to that of saposin C.  
     
     
         16 ) A therapeutic phospholipid composition comprising: 
 a) an anionic phospholipid;    b) a safe and effective amount of the pharmaceutical agent contained within the aqueous interior of the phospholipids; and    c) a fusogenic protein or polypeptide derived from prosaposin;    in a pharmaceutically acceptable carrier, wherein the fusogenic protein or polypeptide is present in a sufficient concentration to deliver the pharmaceutical agent through a biological membrane and the fusogenic protein or polypeptide is associated with the phospholipid through an electrostatic and hydrophobic interaction.    
     
     
         17 ) The therapeutic phospholipid composition of  claim 16  wherein the concentration of anioinic phospholipids is in at least a 10-fold excess, by weight, to that of the fusogenic protein or polypeptide.  
     
     
         18 ) The therapeutic phospholipid composition of  claim 17  wherein the pH of the composition is between about 5.5 and 2.  
     
     
         19 ) The therapeutic phospholipid composition of  claim 18  wherein the anionic phospholipid is an anionic liposome.  
     
     
         20 ) The therapeutic phospholipid composition of  claim 19  wherein the biological membrane is selected from the group consisting of dermal and mucosal membranes.  
     
     
         21 ) The therapeutic phospholipid composition of  claim 20  wherein the fusogenic protein or polypeptide is selected from the group consisting of saposin A, and saposin C, and mixtures thereof.  
     
     
         22 ) The therapeutic phospholipid composition of  claim 20  wherein the fusogenic protein or polypeptide is saposin C.  
     
     
         23 ) The therapeutic phospholipid composition of  claim 20  wherein the fusogenic protein or polypeptide is SEQ. ID. NO. 1.  
     
     
         24 ) The therapeutic phospholipid composition of  claim 20  wherein the fusogenic protein or polypeptide is SEQ. ID. NO. 2.  
     
     
         25 ) The therapeutic phospholipid composition of  claim 20  wherein the fusogenic protein or polypeptide is selected from the group consisting of those proteins or polypeptides given by SEQ ID Nos. 3-6.  
     
     
         26 ) The therapeutic phospholipid composition of  claim 21  wherein the composition is formulated as part of a transdermal patch.  
     
     
         27 ) The therapeutic phospholipid composition of  claim 21  wherein the composition is formulated for enteral or topical administration.  
     
     
         28 ) An anionic liposomal composition used to deliver a pharmaceutical agent through either a dermal or mucosal membrane, wherein the composition comprises: 
 a) anionic liposomes;    b) a safe and effective amount of the pharmaceutical agent contained within the aqueous interior of the liposomes; and    c) saposin C;    in a pharmaceutically acceptable carrier where the pH of the composition is between about 5.5 and 2, wherein the concentration of the saposin C is of a sufficient amount to deliver the pharmaceutical agent through a biological membrane and the saposin C is associated with the surface of the liposomes through an electrostatic and hydrophobic interaction.    
     
     
         29 ) The anionic liposomal composition of  claim 28  wherein the concentration of the anionic liposomes is in at least a 10-fold excess, by weight, to that of saposin C.  
     
     
         30 ) A composition comprising a safe and effective amount of a pharmaceutical agent contained in anionic liposomes, which are associated with a prosaposin-derived fusogenic protein or polypeptide via an electrostatic and hydrophobic interaction, wherein the concentration of the fusogenic protein or polypeptide is of a sufficient amount to deliver the pharmaceutical agent through a biological membrane, the composition contained in a pharmaceutically acceptable carrier, wherein the pH of the composition is between about 5.5 and 2.  
     
     
         31 ) The composition of  claim 30  wherein the concentration of anionic liposomes is in at least a 10-fold excess, by weight, to that of the fusogenic protein or polypeptide.  
     
     
         32 ) The composition of  claim 31  wherein the biological membrane is selected from the group consisting of dermal and mucosal membranes.  
     
     
         33 ) The composition of  claim 32  wherein the fusogenic protein or polypeptide is selected from the group consisting of saposin A, saposin C, and mixtures thereof.  
     
     
         34 ) The composition of  claim 31  wherein the fusogenic protein or polypeptide is saposin C.  
     
     
         35 ) The composition of  claim 31  wherein the fusogenic protein or polypeptide is SEQ. ID. NO. 1.  
     
     
         36 ) The composition of  claim 31  wherein the fusogenic protein or polypeptide is SEQ. ID. NO. 2.  
     
     
         37 . The composition of  claim 31  wherein the fusogenic protein or polypeptide is selected from the group consisting of those proteins or polypeptides given by SEQ ID Nos. 3-6.  
     
     
         38 ) A phospholipid composition used to deliver a pharmaceutical agent through either a dermal or mucosal membrane, wherein the composition comprises: 
 a) anionic liposomes;    b) a safe and effective amount of the pharmaceutical agent contained within the aqueous interior of the liposomes; and    c) saposin C;    in a pharmaceutically acceptable carrier, wherein the pH of the composition is between about 5.5 and 2, the concentration of the saposin C is of a sufficient amount to deliver the pharmaceutical agent through the membrane and the saposin C is associated with the surface of the liposome through an electrostatic and hydrophobic interaction.    
     
     
         39 ) The phospholipid composition of  claim 38  wherein the concentration of the anionic liposomes is in at least a 10-fold excess, by weight, to that of saposin C.  
     
     
         40 ) The polypeptide consisting of SEQ. ID. NO. 1.  
     
     
         41 ) The polypeptide consisting of SEQ. ID. NO. 2.  
     
     
         42 . A compound of the formula consisting of SEQ ID Nos. 3-6.  
     
     
         43 ) A method for treating Gauchers Disease wherein the method comprises the administration of a composition comprising: 
 a) anionic liposomes;    b) a safe and effective amount of acid beta-glucosidase contained within the aqueous interior of the liposomes; and    c) saposin C;    in a pharmaceutically acceptable carrier, wherein the pH of the composition between about 5.5 and 2, the concentration of the saposin C is of a sufficient amount to deliver the pharmaceutical agent through the membrane and the saposin C is associated with the surface of the liposome through an electrostatic and hydrophobic interaction.    
     
     
         44 ) The method of  claim 43  wherein the concentration of the liposome is in at least a 10-fold excess, by weight, to that of saposin C.