Protein and peptide delivery to mammalian cells in vitro

Abstract

Compositions and methods for delivery of proteins and peptides to mammalian cells in vitro are described. Specifically, polypeptide-surfactant complexes formed from noncovalent hydrophobation of polypeptides and reversible hydrophobic modification of polypeptides are described. The compositions can be used to delivery positively charged, negatively charged and charge neutral polypeptides to cells.

We claim:  
     
         1 . A composition for intracellular delivery of a polypeptide, comprising: a dried polypeptide-surfactant complex wherein the surfactant is associated with the polypeptide via a noncovalent bond.  
     
     
         2 . The composition of  claim 1  wherein the surfactant contains a hydrophobic alkyl chain of 4 to 30 carbon atoms.  
     
     
         3 . The composition of  claim 2  wherein the surfactant additionally contains a functional group selected from the list consisting of: membrane active compounds, cell penetrating compounds, cell targeting signals, interaction modifiers, steric stabilizers.  
     
     
         4 . The composition of  claim 1  wherein the complex is associated with one or more lipids.  
     
     
         5 . The composition of  claim 4  wherein the composition consists of a liposome.  
     
     
         6 . The composition of  claim 4  wherein the one or more of the lipids contains a functional group selected from the list consisting of: membrane active compounds, cell penetrating compounds, cell targeting signals, interaction modifiers, steric stabilizers.  
     
     
         7 . The composition of  claim 2  wherein the complex is dissolved in an organic or organic/aqueous solvent.  
     
     
         8 . The composition of  claim 7  wherein the dissolved complex is added to one or more lipids.  
     
     
         9 . A composition for intracellular delivery of a polypeptide, comprising: a polypeptide-surfactant complex wherein the surfactant is associated with the polypeptide via a covalent bond.  
     
     
         10 . The composition of  claim 9  wherein the surfactant contains a functional group selected from the list consisting of: membrane active compounds, cell penetrating compounds, cell targeting signals, interaction modifiers, steric stabilizers.  
     
     
         11 . The composition of  claim 9  wherein the complex is dehydrated.  
     
     
         12 . The composition of  claim 9  wherein the complex is associated with one or more lipids.  
     
     
         13 . The composition of  claim 12  wherein the lipids form a liposome.  
     
     
         14 . The composition of  claim 12  wherein the complex additionally contains a functional group selected from the list consisting of: membrane active compounds, cell penetrating compounds, cell targeting signals, interaction modifiers, steric stabilizers.  
     
     
         15 . The composition of  claim 9  wherein the surfactant consists of an alkyl chlorosilane.  
     
     
         16 . The composition of  claim 15  wherein the silane is selected from the group consisting of compounds of general formula:  
       
         
           
           
               
               
           
         
       
       wherein R 1 , R 2 , and R 3  are independent and are selected from the group consisting of halogen, alkyl, and aryl.  
     
     
         17 . The composition of  claim 9  wherein the surfactant consists of a surfactant-chelator.  
     
     
         18 . The composition of  claim 17  wherein the surfactant-chelator is selected from the list consisting of: molecules of general formula I  
       
         
           
           
               
               
           
         
       
       and molecules of general formula II  
       
         
           
           
               
               
           
         
       
       wherein R is an alkyl group.  
     
     
         19 . The composition of  claim 9  wherein the surfactant consists of an amphipathic maleic anhydride derivative.  
     
     
         20 . A process for the reversible hydrophobic modification of a polypeptide, comprising: forming a polypeptide-surfactant complex wherein the surfactant is selected from the list consisting alkyl chlorosilane, surfactant-chelator and amphipathic maleic anhydride.  
     
     
         21 . A process for delivering a polypeptide to a cell comprising: 
 a) associating a polypeptide with a surfactant via noncovalent interaction to form a polypeptide-surfactant complex;    b) dehydrating the complex to form a polypeptide-surfactant dried salt complex;    c) dissolving the dried salt complex with an organic or organic/aqueous solvent; and,    d) contacting the cell with the dissolved complex of step c).    
     
     
         22 . The process of  claim 21  wherein the one or more lipids are added to the dissolved complex prior to contacting the cell with the complex.  
     
     
         23 . The process of  claim 22  wherein the dissolved complex consists of a liposome.  
     
     
         24 . The process of  claim 22  wherein the dissolved complex is dried and rehydrated in aqueous solvent.