Protocells and their use for targeted delivery of multicomponent cargos to cancer cells

Abstract

Various embodiments provide materials and methods for synthesizing protocells for use in targeted delivery of cargo components to cancer cells. In one embodiment, the lipid bilayer can be fused to the porous particle core to form a protocell. The lipid bilayer can be modified with targeting ligands or other ligands to achieve targeted delivery of cargo components that are loaded within the protocell to a target cell, e.g., a type of cancer. Shielding materials can be conjugated to the surface of the lipid bilayer to reduce undesired non-specific binding.

1 - 28 . (canceled) 
     
     
         29 . A protocell compound comprising:
 a porous core loaded with one or more cargos and surrounded by a lipid bilayer, the one or more cargos selected from the group consisting of peptides, proteins, antibodies, nucleic acids, and drugs; and   a fusogenic ligand.   
     
     
         30 . The protocell compound of  claim 29 , wherein the fusogenic ligand is a fusogenic peptide. 
     
     
         31 . The protocell compound of  claim 29 , wherein the fusogenic ligand is attached to the lipid bilayer. 
     
     
         32 . The protocell compound of  claim 29 , wherein the protocell further comprises a targeting ligand. 
     
     
         33 . The protocell compound of  claim 32 , wherein the targeting ligand is attached to the lipid bilayer. 
     
     
         34 . The protocell compound of  claim 32 , wherein the protocell comprises 1 to about 2048 copies of the targeting ligand. 
     
     
         35 . The protocell of  claim 32 , wherein the targeting ligand is a peptide, a monoclonal antibody, an aptamer, or a vitamin. 
     
     
         36 . The protocell compound of  claim 32 , wherein the targeting ligand binds a cancer cell. 
     
     
         37 . The protocell compound of  claim 36 , wherein the cancer cell is a hepatocellular cancer cell. 
     
     
         38 . The protocell compound of  claim 29 , wherein the protocell further comprises a shielding material. 
     
     
         39 . The protocell compound of  claim 38 , wherein the shielding material is attached to the lipid bilayer. 
     
     
         40 . The protocell compound of  claim 38 , wherein the shielding material comprises polyethylene glycol or cholesterol. 
     
     
         41 . The protocell compound of  claim 29 , wherein the porous core comprises polystyrene, silica, alumina, titania, or zirconia. 
     
     
         42 . The protocell compound of  claim 29 , wherein the porous core comprises silica. 
     
     
         43 . The protocell compound of  claim 29 , wherein the porous core is about 30 nm to about 3000 nm in diameter. 
     
     
         44 . The protocell compound of  claim 29 , wherein the porous core is about 200 nm or less in diameter. 
     
     
         45 . The protocell compound of  claim 29 , wherein the core is spherical, cylindrical, or oblong. 
     
     
         46 . The protocell compound of  claim 29 , wherein the porous core comprises pores with an average pore size of about 2 nm to about 30 nm. 
     
     
         47 . The protocell compound of  claim 29 , wherein the porous core has a porosity of about 25% to about 75%. 
     
     
         48 . The protocell compound of  claim 29 , wherein the porous core has a surface area of about 100 m 2 /g to about 1000 m 2 /g. 
     
     
         49 . The protocell compound of  claim 29 , wherein the porous core is negatively charged. 
     
     
         50 . The protocell compound of  claim 29 , wherein the porous core is positively charged. 
     
     
         51 . The protocell compound of  claim 29 , wherein the porous core comprises an amine-containing compound. 
     
     
         52 . The protocell compound of  claim 29 , wherein the porous core comprises about 0 wt % to about 30 wt % amine-containing compound. 
     
     
         53 . The protocell compound of  claim 29 , wherein the lipid bilayer is a supported lipid bilayer. 
     
     
         54 . The protocell compound of  claim 29 , wherein the lipid bilayer comprises a phospholipid. 
     
     
         55 . The protocell compound of  claim 29 , wherein the lipid bilayer comprises 1,2-dioleoyl-sn-glycero-3-[phosphor-L-serine] (DOPS), 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), PEG-2000-PE (18:1), 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine (DPPE), or cholesterol. 
     
     
         56 . The protocell compound of  claim 29 , wherein the lipid bilayer comprises a negatively-charged lipid, a positively-charged lipid, or a zwitterionic lipid. 
     
     
         57 . The protocell compound of  claim 29 , wherein the lipid bilayer comprises a positively-charged lipid. 
     
     
         58 . The protocell compound of  claim 29 , wherein the one or more encapsulated cargos comprises DNA or RNA. 
     
     
         59 . The protocell compound of  claim 29 , wherein the one or more encapsulated cargos is modified with a nuclear localization sequence. 
     
     
         60 . The protocell compound of  claim 29 , wherein the one or more encapsulated cargos comprises a chemotherapeutic drug. 
     
     
         61 . The protocell compound of  claim 29 , wherein the protocell has a net positive surface charge or a net negative surface charge. 
     
     
         62 . A protocell compound comprising:
 a porous core loaded with one or more cargos and surrounded by a lipid bilayer, the porous core comprising an amine-containing composition;   wherein the one or more cargos are selected from the group consisting of peptides, proteins, antibodies, nucleic acids, and drugs.   
     
     
         63 . A protocell compound comprising:
 a positively-charged porous core loaded with one or more cargos and surrounded by a lipid bilayer; and   wherein the one or more cargos are selected from the group consisting of peptides, proteins, antibodies, nucleic acids, and drugs.   
     
     
         64 . A method of delivering a cargo to a cell comprising contacting the cell with the protocell compound according to  claim 29 . 
     
     
         65 . A method of delivering a cargo to a cell comprising contacting the cell with the protocell compound according to  claim 62 . 
     
     
         66 . A method of delivering a cargo to a cell comprising contacting the cell with a protocell compound according to  claim 63 . 
     
     
         67 . A method of treating a disease comprising administering to a subject a plurality of protocell compounds according to  claim 29 . 
     
     
         68 . A method of treating a disease comprising administering to a subject a plurality of protocell compounds, the protocell compounds according to  claim 62 . 
     
     
         69 . A method of treating a disease comprising administering to a subject a plurality of protocell compounds according to  claim 63 . 
     
     
         70 . A method of forming a protocell compound of  claim 29  comprising:
 surrounding the porous core with the lipid bilayer, and 
 attaching a fusogenic ligand to the lipid bilayer. 
 
     
     
         71 . A method of forming a protocell compound of  claim 62  comprising surrounding the porous core with the lipid bilayer. 
     
     
         72 . A method of forming a protocell compound of  claim 63  comprising surrounding the porous core with the lipid bilayer.