Emulsion and micellar formulations for the delivery of biologically active substances to cells

Abstract

New emulsion and micelle formulations are described as are complexes of these formulations with biologically active substances. The novel formulations are different from cationic lipid vectors such as cationic liposomes in that the complexes formed between biologically active substances and the emulsion and micellar formulations of this invention are physically stable and their transfection activity is resistant to the presence of serum. These novel formulations are disclosed to be useful in areas such as gene therapy or vaccine delivery.

1 . An oil-in-water emulsion formulation comprising lipid components and an aqueous carrier, wherein the lipid components comprise an oil component, a cationic amphiphile component and a nonionic surfactant component.  
     
     
         2 . The emulsion formulation of  claim 1  wherein the lipid components further comprise a neutral phospholipid component.  
     
     
         3 . The emulsion formulation of  claim 1  wherein the oil component is present in an amount from about 10 to about 80 weight % of the total lipid components in the formulation, the amphiphile component is present in an amount from about 5 to about 80 weight % of the total lipid components and the nonionic surfactant component is present in an amount from about 5 to about 50 weight % of the total lipid components.  
     
     
         4 . The emulsion formulation of  claim 3  further comprising a neutral phospholipid component present in an amount from about 5 to about 25 weight % of the total lipid components in the formulation.  
     
     
         5 . The emulsion formulation of  claim 4 , wherein the amphiphile component is a cationic lipid.  
     
     
         6 . A micellar formulation comprising lipid components and an aqueous carrier, wherein the lipid components comprise a cationic amphiphile component and a nonionic surfactant component.  
     
     
         7 . The micellar formulation of  claim 6 , wherein the lipid components further comprise a neutral phospholipid component.  
     
     
         8 . The micellar formulation of  claim 6  wherein the amphiphile component is present in an amount from about 10 to about 90 weight % of the total lipid components and the nonionic surfactant component is present in an amount from about 90 to about 10 weight % of the total lipid components.  
     
     
         9 . The micellar formulation of  claim 8 , further comprising a neutral phospholipid component present in an amount from about 5 to about 40 weight % of the total lipid components.  
     
     
         10 . The micellar formulation of  claim 9 , wherein the amphiphile component is a cationic lipid.  
     
     
         11 . A complex for facilitating the delivery of a negatively charged biologically active substance to cells, said complex comprising the negatively charged biologically active substance and the emulsion formulation of  claim 1 .  
     
     
         12 . The complex of  claim 11 , wherein the negatively charged substance is a nucleic acid.  
     
     
         13 . The complex of  claim 12 , wherein the weight ratio of nucleic acid to total lipid components in the emulsion formulation in said complex is about 1:1 to about 1:50.  
     
     
         14 . A complex for facilitating the delivery of a negatively charged biologically active substance to cells, said complex comprising said negatively charged substance and the micellar formulation of  claim 6 .  
     
     
         15 . The complex of  claim 14 , wherein the negatively charged substance is a nucleic acid.  
     
     
         16 . The complex of  claim 15 , wherein the weight ratio of nucleic acid to total lipid components in the micellar formulation in said complex is from about 1:1 to about 1:50.  
     
     
         17 . A method for delivering a negatively charged biologically active substance to cells comprising exposing the cells to the complex of  claim 11  thereby facilitating the delivery of the negatively charged biologically active substance to the cells.  
     
     
         18 . The method of  claim 17  wherein the cells are mammalian cells exposed to the complex in the presence of serum.  
     
     
         19 . A method for delivering a negatively charged biologically active substance to cells comprising exposing the cells to the complex of  claim 14  thereby facilitating the delivery of the negatively charged biologically active substance to the cells.  
     
     
         20 . The method of  claim 19  wherein the cells are mammalian cells exposed to the complex in the presence of serum.  
     
     
         21 . The methods for delivering a negatively charged biologically active substance to cells according to claims  17  and  19  wherein the cells are exposed to the complex in vivo by administering the complexes to an animal or human in an amount effective to facilitate the delivery of the negatively charged substance to the cells of the animal or human.  
     
     
         22 . The method of  claim 21 , wherein the negatively charged substance is a nucleic acid.  
     
     
         23 . The methods for delivering a negatively charged biologically active substance to cells according to claims  17  and  19  wherein the cells are exposed to the complex in vitro.  
     
     
         24 . The method of  claim 23 , wherein the negatively charged substance is a nucleic acid.  
     
     
         25 . A method of producing an oil-in-water emulsion formulation comprising: 
 (a) combining an oil component, a cationic amphiphile component and a nonionic surfactant component; and    (b) adding aqueous carrier to produce said emulsion formulation.    
     
     
         26 . A method of producing a micellar formulation comprising: 
 (a) combining a cationic amphiphile component and a nonionic surfactant component; and    (b) adding aqueous carrier to produce said micellar formulation.    
     
     
         27 . The methods of claims  25  and  26  further comprising combining the components of step (a) with a neutral phospholipid component.  
     
     
         28 . The method of  claim 27 , wherein the components of step (a) are combined in an organic solvent and the solvent is removed to leave a lipid film prior to step (b).  
     
     
         29 . A method of producing a lipid film having an oil component, a cationic amphiphile component and a nonionic surfactant component; said method comprising: 
 (a) combining an organic solvent with the oil component, the amphiphile component and the nonionic surfactant component; and    (b) removing the organic solvent to leave said lipid film.    
     
     
         30 . A method of producing a lipid film having a cationic amphiphile component and a nonionic surfactant component; said method comprising: 
 (a) combining an organic solvent with the amphiphile component and the nonionic surfactant component; and    (b) removing the organic solvent to leave said lipid film.    
     
     
         31 . A lipid film capable of forming an oil-in-water emulsion upon suspension in an aqueous carrier, said film having an oil component, a cationic amphiphile component and a nonionic surfactant component.  
     
     
         32 . A lipid film capable of forming a micelle upon suspension in solution, said film having a cationic amphiphile component and a nonionic surfactant component.  
     
     
         33 . The lipid films of claims  32  and  33 , said films further having a neutral phospholipid component.