Compositions and methods for modified dendrimer nanoparticle delivery

Abstract

Compositions and methods for modified dendrimer nanoparticle (“MDNP”) delivery of therapeutic, prophylactic and/or diagnostic agent such as large repRNA molecules to the cells of a subject have been developed. MDNPs efficiently drive proliferation of antigen-specific T cells against intracellular antigen, and potentiate antigen-specific antibody responses. MDNPs can be multiplexed to deliver two or more different repRNAs to modify expression kinetics of encoded antigens and to simultaneous deliver repRNAs and mRNAs including the same UTR elements that promote expression of encoded antigens.

1 - 33 . (canceled) 
     
     
         34 . A modified dendrimer nanoparticle comprising:
 one or more zero to seven generation alkylated dendrimers; and   one or more amphiphilic polymers;   wherein the alkylated dendrimer is not a poly(amido amine) dendrimer;   wherein the amphiphilic polymer is not a dendrimer; and   wherein if the nanoparticle comprises cholesterol, it is present only as part of the amphiphilic polymer;   the nanoparticles containing one or more nucleic acids that express and/or encode one or more therapeutic, prophylactic, and/or diagnostic agents; and optionally,   one or more therapeutic, prophylactic and/or diagnostic agents.   
     
     
         35 . The nanoparticle of  claim 34 , wherein the alkylated dendrimer is selected from the group consisting of poly(ethylenimine), polyester, polylysine, polypropyleneimine, diaminobutane amine polypropylenimine tetramine, polypropylamine, iptycene, aliphatic polyether, and aromatic polyether dendrimers. 
     
     
         36 . The nanoparticle of  claim 34 , wherein the alkylated dendrimer is a poly(ethylenimine) dendrimer. 
     
     
         37 . The nanoparticle of  claim 34 , wherein the alkylated dendrimer is a diaminobutane amine polypropylenimine tetramine dendrimer. 
     
     
         38 . The nanoparticle of  claim 34 , wherein the alkylated dendrimer is selected from the group consisting of generation 0 dendrimers, generation 1 dendrimers, generation 2 dendrimers, and generation 3 dendrimers. 
     
     
         39 . The nanoparticle of  claim 34 , wherein the alkylated dendrimer is alkylated by epoxide-terminated alkyl chains, in a size between 1 and 30 carbons, inclusive. 
     
     
         40 . The nanoparticle of  claim 39 , wherein the epoxide-terminated alkyl chains are between 6 and 16 carbons, inclusive. 
     
     
         41 . The nanoparticle of  claim 34 , wherein the amphiphilic polymer comprises a hydrophilic component selected from the group consisting of polyalkylene oxides and block copolymers thereof. 
     
     
         42 . The nanoparticle of  claim 34 , wherein the amphiphilic polymer comprises polyalkylene oxide. 
     
     
         43 . The nanoparticle of  claim 42 , wherein the polyalkylene oxide is 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000]. 
     
     
         44 . The nanoparticle of  claim 34 , wherein the amphiphilic polymer comprises polyethylene glycol with a molecular weight between 120 Da and 25,000 Da. 
     
     
         45 . The nanoparticle of  claim 44 , wherein the polyethylene glycol has a molecular weight of 2,000 Da. 
     
     
         46 . The nanoparticle of  claim 34 , wherein the amphiphilic polymer comprises a hydrophobic component selected from the group consisting of a lipid and a phospholipid. 
     
     
         47 . The nanoparticle of  claim 41 , wherein the mass ratio of the alkylated dendrimer to the hydrophilic component of the amphiphilic polymer is between 20:1 and 5:1, inclusive. 
     
     
         48 . The nanoparticle of  claim 34 , comprising a therapeutic, prophylactic or diagnostic agent selected from the group consisting of proteins, peptides, carbohydrates, nucleic acids, lipids, small molecules, and combinations thereof. 
     
     
         49 . The nanoparticle of  claim 34 , wherein the nucleic acids are selected from the group consisting of complementary DNA (cDNA), replicating RNA (repRNA), messenger RNA (mRNA), small interfering RNA (siRNA), transfer RNA (tRNA), guide-strand RNA (sgRNA), microRNA (miRNA), double-stranded RNA (dsRNA) that is at least 24 nucleotides in length, and combinations thereof. 
     
     
         50 . The nanoparticle of  claim 49 , wherein the nucleic acids comprise one or more ribonucleic acid sequences of between approximately 10 and 20,000 bases in length, inclusive. 
     
     
         51 . The nanoparticle of  claim 50 , wherein the ribonucleic acid sequences are between approximately 9,000 and 15,000 bases in length, inclusive. 
     
     
         52 . The nanoparticle of  claim 49 , wherein the mass ratio of the alkylated dendrimer to ribonucleic acid is between 10:1 and 1.5:1, inclusive. 
     
     
         53 . The nanoparticle of  claim 52 , wherein the mass ratio of the alkylated dendrimer to ribonucleic acid is 5:1. 
     
     
         54 . The nanoparticle of  claim 49 , wherein the replicating RNA comprises a modified alphavirus replicon RNA. 
     
     
         55 . The nanoparticle of  claim 49 , wherein the nanoparticle comprises two or more replicating RNAs, wherein the replicating RNAs independently encode one or more antigens from an infectious agent, parasite, or abnormal proliferation disorder. 
     
     
         56 . The nanoparticle of  claim 55 , wherein the abnormal proliferation disorder is cancer. 
     
     
         57 . The nanoparticle of  claim 49 , wherein the nanoparticle comprises a replicating RNA and a messenger RNA, and wherein the messenger RNA is modified to include the 5′ and 3′ untranslated regions of the replicating RNA. 
     
     
         58 . The nanoparticle of  claim 34 , wherein the size of the nanoparticle is between 30 nm and 450 nm. 
     
     
         59 . The nanoparticle of  claim 58 , wherein the size of the nanoparticle is between 60 nm and 250 nm, inclusive. 
     
     
         60 . A method for delivering one or more nucleic acids to a subject, comprising administering to the subject the nanoparticle of  claim 34 . 
     
     
         61 . The method of  claim 60 , wherein the nanoparticle delivers one or more replicating RNAs, and wherein the replicating RNAs express one or more proteins in the cells of the subject. 
     
     
         62 . The method of  claim 61 , wherein the one or more replicating RNAs express two or more proteins in the cells of the subject, and wherein the replicating RNAs express the proteins at different rates.