Lipid Particle Formulations for Delivery of RNA and Water-Soluble Therapeutically Effective Compounds to a Target Cell

Abstract

The present invention relates to lipid particles comprising at least one cationic lipid, at least one water-soluble therapeutically effective compound and RNA. Further, the present invention relates to a pharmaceutical composition comprising such particles. Said pharmaceutical composition is useful for inducing an immune response. It is also useful in a prophylactic and/or therapeutic treatment of a disease involving an antigen. Furthermore, the present invention relates to a method for producing the particles.

1 . A lipid particle comprising:
 (i) at least one cationic and/or pH responsive lipid,   (ii) at least one water-soluble therapeutically effective compound, and   (iii) RNA.   
     
     
         2 . The particle of  claim 1  wherein the lipid forms a structure receiving the at least one water soluble therapeutically effective compound and the RNA. 
     
     
         3 . The particle of  claim 1  or  2  which comprises a lamellar internal organization. 
     
     
         4 . The particle of any one of  claims 1  to  3 , wherein the lamellar internal organization comprises 2 to 40, preferably 2 to 20, more preferably 2 to 10, in particular 3 to 8 lamellae per row. 
     
     
         5 . The particle of any one of  claims 1  to  4 , wherein the RNA is pharmaceutically active or encodes at least one pharmaceutically active peptide or protein. 
     
     
         6 . The particle of any one of  claims 1  to  5 , wherein the RNA encodes at least one antigen. 
     
     
         7 . The particle of  claim 6 , wherein the antigen is a disease-associated antigen or elicits an immune response against a disease-associated antigen or cells expressing a disease-associated antigen. 
     
     
         8 . The particle of any one of  claims 1  to  7 , wherein the therapeutically effective compound has a molecular mass lower than 1000 Da. 
     
     
         9 . The particle of any one of  claims 1  to  8 , wherein the therapeutically effective compound is useful in immunotherapy. 
     
     
         10 . The particle of any one of  claims 1  to  9 , wherein the therapeutically effective compound is an agent stimulating γδ T cells, preferably Vγ9Vδ2 T cells. 
     
     
         11 . The particle of  claim 10 , wherein the agent stimulating γδ T cells is a bisphosphonate, preferably a nitrogen-containing bisphosphonate (aminobisphosphonate). 
     
     
         12 . The particle of  claim 10  or  11 , wherein the agent stimulating γδ T cells is selected from the group consisting of zoledronic acid, clodronic acid, ibandronic acid, pamidronic acid, risedronic acid, minodronic acid, olpadronic acid, alendronic acid, incadronic acid and salts thereof. 
     
     
         13 . The particle of any one of  claims 1  to  12 , which comprises at least one helper lipid. 
     
     
         14 . The particle of  claim 13 , wherein the helper lipid is a neutral lipid or negatively charged lipid. 
     
     
         15 . The particle of any one of  claims 1  to  14 , wherein the at least one cationic lipid comprises 1,2-dimyristoyl-sn-glycero-3-ethylphosphocholine (DMEPC), 1,2-di-O-octadecenyl-3-trimethylammonium propane (DOTMA) and/or 1,2-dioleoyl-3-trimethylammonium propane (DOTAP). 
     
     
         16 . The particle of any one of  claims 13  to  15 , wherein the at least one helper lipid comprises 1,2-di-(9Z-octadecenoyl)-sn-glycero-3-phosphoethanolamine (DOPE), cholesterol (Chol), 1-palmitoyl-2-oleoyl-sn-glycero-3phosphocholin (POPC) and/or 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC). 
     
     
         17 . The particle of any one of  claims 1  to  16 , wherein the particle has an average diameter in the range of from about 50 nm to about 1000 nm. 
     
     
         18 . The particle of  claim 17 , wherein the particle has an average diameter
 (i) in the range of from about 50 nm to about 400 nm, preferably from about 50 nm to about 200 nm, or   (ii) in the range of from about 200 nm to about 1000 nm, preferably from about 200 nm to about 800 nm, more preferably from about 300 nm to about 600 nm.   
     
     
         19 . The particle of any one of  claims 1  to  18 , which is obtainable by addition of the RNA to a colloidal lipid dispersion comprising the at least one cationic lipid and the at least one water-soluble therapeutically effective compound. 
     
     
         20 . The particle of  claim 19  wherein the colloidal lipid dispersion comprising the at least one cationic lipid and the at least one water-soluble therapeutically effective compound is obtainable by injection of an ethanol solution of the lipids into an aqueous phase comprising the at least one water-soluble therapeutically effective compound. 
     
     
         21 . A pharmaceutical composition comprising particles as set forth in any one of  claims 1  to  20 . 
     
     
         22 . The pharmaceutical composition of  claim 21 , wherein, after systemic administration of the particles, at least a portion of the RNA and at least a portion of the therapeutically effective compound are delivered to a target cell, preferably to the same target cell. 
     
     
         23 . The pharmaceutical composition of  claim 22 , wherein at least a portion of the RNA and at least a portion of the therapeutically effective compound are delivered to the cytosol of the target cell. 
     
     
         24 . The pharmaceutical composition of  claim 22  or  23 , wherein the RNA is RNA encoding a peptide or protein and the RNA is translated by the target cell to produce the peptide or protein. 
     
     
         25 . The pharmaceutical composition of any one of  claims 22  to  24 , wherein the target cell is a spleen cell, preferably an antigen presenting cell, more preferably a professional antigen presenting cell, more preferably a dendritic cell. 
     
     
         26 . The pharmaceutical composition of any one of  claims 21  to  25 , wherein, after systemic administration of the particles, RNA accumulation and/or RNA expression in the spleen occurs. 
     
     
         27 . The pharmaceutical composition of any one of  claims 21  to  26 , wherein, after systemic administration of the particles, no or essentially no RNA accumulation and/or RNA expression in the lung and/or liver occurs. 
     
     
         28 . The pharmaceutical composition of any one of  claims 21  to  27 , wherein, after systemic administration of the particles, RNA accumulation and/or RNA expression in the spleen is at least 5-fold the amount of RNA accumulation and/or RNA expression in the lung and/or liver. 
     
     
         29 . The pharmaceutical composition of any one of  claims 21  to  28 , wherein, after systemic administration of the particles, RNA accumulation and/or RNA expression in antigen presenting cells, preferably professional antigen presenting cells in the spleen occurs. 
     
     
         30 . The pharmaceutical composition of  claim 29 , wherein the antigen presenting cells are dendritic cells and/or macrophages. 
     
     
         31 . The pharmaceutical composition of any one of  claims 22  to  30 , wherein systemic administration is by parenteral administration, preferably by intravenous administration, subcutaneous administration, intradermal administration or intraarterial administration. 
     
     
         32 . The pharmaceutical composition of any one of  claims 21  to  31 , wherein the composition further comprises one or more pharmaceutically acceptable carriers, diluents and/or excipients. 
     
     
         33 . The pharmaceutical composition of any one of  claims 21  to  32 , wherein the composition further comprises at least one adjuvant. 
     
     
         34 . The pharmaceutical composition of any one of  claims 21  to  33 , wherein the composition is formulated for systemic administration. 
     
     
         35 . The pharmaceutical composition of any one of  claims 21  to  34  for inducing or enhancing an immune response, preferably an immune response against cancer. 
     
     
         36 . The pharmaceutical composition of any one of  claims 21  to  35  for use in a prophylactic and/or therapeutic treatment of a disease involving an antigen, preferably a cancer disease. 
     
     
         37 . A method for delivering an antigen to antigen presenting cells, preferably professional antigen presenting cells, in the spleen, or expressing an antigen in antigen presenting cells, preferably professional antigen presenting cells, in the spleen comprising administering to a subject a pharmaceutical composition of any one of  claims 21  to  34 . 
     
     
         38 . The method of  claim 37 , wherein the antigen presenting cells are dendritic cells and/or macrophages. 
     
     
         39 . A method for inducing or enhancing an immune response, preferably an immune response against cancer, in a subject comprising administering to the subject a pharmaceutical composition of any one of  claims 21  to  34 . 
     
     
         40 . A method for stimulating, priming and/or expanding T cells in a subject comprising administering to the subject a pharmaceutical composition of any one of  claims 21  to  34 . 
     
     
         41 . A method of treating or preventing a disease involving an antigen, preferably a cancer disease, in a subject comprising administering to the subject a pharmaceutical composition of any one of  claims 21  to  34 . 
     
     
         42 . A method of producing a particle of any one of  claims 1  to  20  comprising the following steps of:
 (i) providing a colloidal lipid dispersion comprising at least one cationic lipid and at least one water-soluble therapeutically effective compound, and 
 (ii) adding RNA to the lipid dispersion comprising at least one cationic lipid and at least one water-soluble therapeutically effective compound. 
 
     
     
         43 . The method of  claim 42  wherein the colloidal lipid dispersion comprising at least one cationic lipid and at least one water-soluble therapeutically effective compound is provided by injection of an ethanol solution of lipids into an aqueous phase comprising the at least one water-soluble therapeutically effective compound. 
     
     
         44 . The method of  claim 42  or  43 , wherein the number of positive charges derived from the cationic lipids divided by the number of negative charges derived from the RNA is between 0.025 and 4.