Lipid nanoparticles

Abstract

The present invention relates to the field of lipid nanoparticles (LNP); more specifically comprising an ionisable lipid, a phospholipid, a sterol, a PEG lipid and one or more nucleic acids. The LNP's of the present invention are characterized in having a minimal average diameter of about 140 nm, thereby inducing a more potent immune response. The present invention provides use of the LNPs for immunogenic delivery of nucleic acid molecules, specifically mRNA; thereby making them highly suitable for use in vaccines, such as for the treatment of cancer or infectious diseases. Finally, methods are provided for preparing such LNP's.

1 . A lipid nanoparticle (LNP) comprising:
 an ionizible lipid;   from about 5 mol % to about 15 mol % of a phospholipid selected from the group consisting of DOPE, DOPC and mixtures thereof;   a sterol;   a PEG lipid; and   one or more nucleic acid molecules;   
       wherein said LNP has a minimal average diameter of about 160 nm to about 230 nm to target the spleen in preference over the liver. 
     
     
         2 . The LNP as defined in  claim 1 , wherein said ionizible lipid is a compound of formula (I): 
       
         
           
           
               
               
           
         
         wherein:
 RCOO is selected from the group consisting of: myristoyl, α-D-Tocopherolsuccinoyl, linoleoyl and oleoyl; and 
 X is selected from the group consisting of: 
 
       
       
         
           
           
               
               
           
         
       
     
     
         3 . The LNP of  claim 1 ; wherein said sterol is selected from the group consisting of cholesterol, ergosterol, campesterol, oxysterol, antrosterol, desmosterol, nicasterol, sitosterol and stigmasterol; preferably cholesterol. 
     
     
         4 . The LNP of  claim 1 ; wherein said PEG lipid is selected from the group consisting of: PEG-modified phosphatidylethanolamine, a PEG-modified phosphatidic acid, a PEG-modified ceramide, a PEG-modified dialkylamine, a PEG-modified diacylglycerol, a PEG-modified dialkylglycerol, and mixtures thereof. 
     
     
         5 . The LNP of  claim 1 ; wherein said LNP comprises about and between 10 mol % and 60 mol % of said ionizable lipid and/or said LNP comprises about and between 15 mol % and 50 mol % of said sterol. 
     
     
         6 . The LNP of  claim 1 ; wherein said one or more nucleic acid molecules are selected from the group consisting of mRNA and DNA. 
     
     
         7 . The LNP of  claim 6 ; wherein said one or more mRNA molecules are selected from the group of immunomodulatory polypeptide-encoding mRNA and/or antigen-encoding mRNA. 
     
     
         8 . The LNP of  claim 6 ; wherein said immunomodulatory-encoding mRNA is selected from the group consisting of mRNA molecules encoding for CD40L, mRNA molecules encoding for CD70, and mRNA molecules encoding for caTLR4. 
     
     
         9 . A pharmaceutical composition or a vaccine comprising one or more of the LNP's of  claim 1  and an acceptable pharmaceutical carrier. 
     
     
         10 . (canceled) 
     
     
         11 . A method for the preparation of a nucleic acid-containing LNP having a minimal average diameter of about 160 nm to about 230 nm, said method comprising:
 preparing a first alcoholic composition comprising an ionizible lipid, a phospholipid, a sterol, a PEG lipid, and a suitable alcoholic solvent;   preparing a second aqueous composition comprising one or more nucleic acids and an aqueous solvent supplemented with an appropriate buffer;   mixing said first and second composition in a in a microfluidic mixing device using the following settings:   
       Total Flow Rate (FR) of about 0.5 to about 8 ml/min and 
       Flow Rate Ratio (FRR) of about 1/1 to 5/1. 
     
     
         12 . The method of  claim 11 , wherein said alcoholic solvent is ethanol and/or said aqueous solvent is water. 
     
     
         13 - 14 . (canceled) 
     
     
         15 . The method of  claim 11 , wherein the FR is 1 ml/min and the FRR is about 3/1. 
     
     
         16 . The method of  claim 15 , wherein the ionizible lipid comprises from about 35 mol % to about 50 mol % of the first alcoholic composition. 
     
     
         17 . The LNP of  claim 1 , wherein the LNP targets the spleen in preference over the liver at a ratio of at least 8 to 1. 
     
     
         18 . The LNP of  claim 1 , wherein the LNP comprises from about 35 mol % to about 50 mol % ionizible lipid.