Porous nanoparticle-supported lipid bilayers (protocells) for targeted delivery and methods of using same

Abstract

The present invention is directed to protocells for specific targeting of hepatocellular and other cancer cells which comprise a nanoporous silica core with a supported lipid bilayer; at least one agent which facilitates cancer cell death (such as a traditional small molecule, a macromolecular cargo (e.g. siRNA or a protein toxin such as ricin toxin A-chain or diphtheria toxin A-chain) and/or a histone-packaged plasmid DNA disposed within the nanoporous silica core (preferably supercoiled in order to more efficiently package the DNA into protocells) which is optionally modified with a nuclear localization sequence to assist in localizing protocells within the nucleus of the cancer cell and the ability to express peptides involved in therapy (apoptosis/cell death) of the cancer cell or as a reporter, a targeting peptide which targets cancer cells in tissue to be treated such that binding of the protocell to the targeted cells is specific and enhanced and a fusogenic peptide that promotes endosomal escape of protocells and encapsulated DNA. Protocells according to the present invention may be used to treat cancer, especially including hepatocellular (liver) cancer using novel binding peptides (c-MET peptides) which selectively bind to hepatocellular tissue or to function in diagnosis of cancer, including cancer treatment and drug discovery.

1 - 90 . (canceled) 
     
     
         91 . A composition comprising a population of protocells comprising a mesoporous silica or metal oxide nanoparticle core surrounded by a lipid bilayer and a cargo, wherein the protocells and the mesoporous silica nanoparticle cores in the population are monodisperse, and wherein the cargo comprises nucleic acid. 
     
     
         92 . The protocell composition according to  claim 91 , wherein the protocells vary no more than about 5% in diameter from a mean diameter. 
     
     
         93 . The protocell composition according to  claim 91 , wherein the protocells have a mean diameter of between about 10 nm and about 500 nm. 
     
     
         94 . The protocell composition according to  claim 91 , wherein the protocells have a mean diameter of between about 20 nm and about 200 nm. 
     
     
         95 . The protocell composition according to  claim 91 , wherein the lipid bilayer comprises cholesterol. 
     
     
         96 . The protocell composition according to  claim 91 , wherein the lipid bilayer comprises a PEG-conjugated lipid. 
     
     
         97 . The protocell composition according to  claim 91 , wherein the protocells further comprise a targeting species attached to the lipid bilayer. 
     
     
         98 . The protocell composition according to  claim 97 , wherein the targeting species is a peptide, an antibody, an antibody fragment, an aptamer, or a carbohydrate. 
     
     
         99 . The protocell composition according to  claim 91 , wherein the protocells further comprise a fusogenic peptide attached to the lipid bilayer. 
     
     
         100 . The protocell composition according to  claim 91 , wherein the protocells further comprise a therapeutic agent, an imaging agent or a diagnostic agent. 
     
     
         101 . The protocell composition according to  claim 91 , wherein the protocells further comprise an anticancer agent or an antiviral agent. 
     
     
         102 . The protocell composition according to  claim 91 , wherein the nucleic acid is DNA or RNA. 
     
     
         103 . The protocell composition according to  claim 102 , wherein the nucleic acid is a double stranded linear DNA, a plasmid DNA, a small interfering RNA, a small hairpin RNA, a microRNA, or supercoiled DNA. 
     
     
         104 . The protocell composition according to  claim 91 , wherein the protocells further comprise
 a polypeptide.   
     
     
         105 . The protocell composition according to  claim 91 , wherein the core has a multimodal pore configuration. 
     
     
         106 . The protocell composition according to  claim 91 , wherein the core comprises an organosilane. 
     
     
         107 . The protocell composition according to  claim 91 , wherein the core comprises an an amine-containing silane, a charged organosilane, a hydrophobic organosilane, or a reactive organosilane. 
     
     
         108 . A pharmaceutical composition comprising the protocell composition according to  claim 91  and a pharmaceutically acceptable excipient. 
     
     
         109 . A method of treating a disease in a subject comprising administering to the subject the protocell composition according to  claim 91 . 
     
     
         110 . The method according to  claim 109 , wherein the disease is cancer.