Compositions of nucleic acid-containing nanoparticles for in vivo delivery

Abstract

Compositions comprising a polymeric micellar nanoparticle composition comprising a block or graft copolymer comprising at least one polycationic polymer and at least one polyethylene glycol (PEG) polymer having an average molecular weight less than 1 kDa, and at least one nucleic acid, wherein the graft or block copolymer and at least one nucleic acid are complexed and condensed into a shaped micellar nanoparticle that is stable in biological media are disclosed. The presently disclosed subject matter also provides a method for preparing the presently disclosed polymeric micellar nanoparticle compositions, a method for targeting at least one metastatic cancer cell in a subject, and a method for treating a disease or condition using the presently disclosed polymeric micellar nanoparticle compositions.

That which is claimed: 
     
         1 . A polymeric micellar nanoparticle composition, comprising:
 (a) a block or graft copolymer comprising at least one polycationic polymer and at least one polyethylene glycol (PEG) polymer having an average molecular weight less than 1 kDa; and   (b) at least one nucleic acid;   wherein the graft or block copolymer and the at least one nucleic acid are complexed and condensed into a shaped micellar nanoparticle that is stable in biological media.   
     
     
         2 . The composition of  claim 1 , wherein:
 (i) the at least one PEG polymer has a molecular weight ranging from about 400 Da to about 1 kDa; or   (ii) the at least one PEG polymer has a molecular weight ranging from about 500 Da to about 700 Da; or   (iii) the copolymer is a graft copolymer and the at least one PEG polymer has a graft density ranging from about 0.25 mol % to about 10 mol %; or   (iv) the at least one PEG polymer is terminated with a functional group selected from the group consisting of a terminal acrylate group, a terminal alkoxy group, a terminal amino group, terminal carboxyl group, a terminal hydroxyl group, a terminal maleimide group, a terminal methacrylate group, a terminal methoxy group, a terminal 2-pyridyldithio (SPDP) group, a terminal thiol group, a negatively charged terminal group, or amphoteric group, or combinations thereof.   
     
     
         3 . The composition of  claim 1 , wherein the shaped micellar nanoparticle is selected from the group consisting of a spherically-shaped micellar nanoparticle, a rod-shaped micellar nanoparticle, and a worm-shaped micellar nanoparticle. 
     
     
         4 . The composition of  claim 1 , further comprising a ligand conjugated to the at least one PEG polymer and/or the functional group. 
     
     
         5 . The composition of  claim 4 , wherein:
 (i) the ligand is selected from the group consisting of a diagnostic agent, an imaging agent, a targeting agent, a theranostic agent, a therapeutic agent, and combinations thereof; or   (ii) the ligand is selected from the group consisting of a DNA, RNA, polypeptide, antibody, antibody fragment, antigen, carbohydrate, protein, peptide, enzyme, amino acid, hormone, steroid, vitamin, drug, virus, polysaccharide, lipid, lipopolysaccharide, glycoprotein, lipoprotein, nucleoprotein, oligonucleotide, immunoglobulin, albumin, hemoglobin, coagulation factor, peptide hormone, protein hormone, non-peptide hormone, interleukin, interferon, cytokine, peptides comprising a tumor-specific epitope, cell, cell-surface molecule, cell adhesion peptide, cell-binding peptide, cell receptor ligand, small organic molecule, small organometallic molecule, nucleic acid, oligonucleotide, transferrin, metabolites thereof, and antibodies or agents that bind to any of the above substances; or   (iii) the ligand is detectable using an imaging modality selected from the group consisting of bioluminescence imaging, fluorescence imaging, magnetic resonance imaging (MRD, positron emission tomography (PET), x-ray computed tomography (CT), single-photon emission computed tomography (SPECT), and combinations thereof; or   (iv) the ligand comprises a peptide comprising the amino acid sequence Ac-CCRRYVVLPRWLC (SEQ ID NO: 1), cyclic RGD-thiol peptide (cRGD), or a peptide comprising the amino acid sequence YIGSR (SEQ ID NO: 3); or   (v) the ligand comprises a moiety that binds to a tumor-specific antigen; or   (vi) the ligand comprises a prostate-specific membrane antigen (PSMA)-binding moiety.   
     
     
         6 . The composition of  claim 1 , wherein:
 (i) the at least one polycationic polymer is selected from the group consisting of linear polyethylenimine (LPEI), poly-lysine, poly-arginine, poly-histidine, chitosan, branched PEI, a poly (beta-aminoester), a polyphosphoester (PPE), and polyphosphoramidate (PPA); or   (ii) the at least one polycationic polymer is LPEI; or   (iii) the at least one polycationic polymer is not branched PEI; or   (iv) the graft copolymer is not a branched PEI(25 kDa)-g-linear PEG(550 Da) n  copolymer, wherein n is the average number of PEG blocks per one PEI macromolecule and n is equal to 35.   
     
     
         7 . The composition of  claim 6 , wherein:
 (i) the LPEI has a molecular weight ranging from about 2 kDa to about 50 kDa; or   (ii) the LPEI has a molecular weight of about 22 kDa.   
     
     
         8 . The composition of  claim 1 , wherein:
 (i) the at least one nucleic acid has a length ranging from about 10 bases to about 10 kilobases (kb); or   (ii) the at least one nucleic acid is selected from the group consisting of an antisense oligonucleotide, cDNA, genomic DNA, guide RNA, plasmid DNA, vector DNA, mRNA, miRNA, piRNA, shRNA, and siRNA; or   (iii) the at least one nucleic acid comprises an expression vector encoding at least one reporter gene operably linked to a promoter; or   (iv) the at least one nucleic acid comprises an expression vector encoding at least one antigen epitope operably linked to a promoter.   
     
     
         9 . The composition of  claim 8 , wherein:
 (i) the reporter gene is selected from the group consisting of a bioluminescent reporter gene, a fluorescent reporter gene, a PET reporter gene, and combinations thereof; or   (ii) the promoter is selected from the group consisting of a constitutively active promoter, an inducible promoter, a tissue-specific promoter, and a tumor-specific promoter; or   (iii) the expression vector further comprises a therapeutic gene; or   (iv) the expression vector further comprises an antigen gene.   
     
     
         10 . The composition of  claim 9 , wherein:
 (i) the therapeutic gene is selected from the group consisting of a cytotoxic gene, an immunomodulator gene, a suicide gene, and a tumor suppressor gene; or   (ii) the antigen gene encodes at least one antigen against infectious diseases, allergens, or cancer cells.   
     
     
         11 . The composition of  claim 1 , further comprising a therapeutic agent or a chemotherapeutic agent. 
     
     
         12 . The composition of  claim 1 , wherein the micellar nanoparticle composition targets at least one target cancer cell. 
     
     
         13 . The composition of  claim 12 , wherein:
 (i) the cancer cell comprises a metastatic cancer cell; or   (ii) the cancer cell is selected from the group consisting of a breast cancer cell, a cervical cancer cell, a melanoma cancer cell, and a prostate cancer cell.   
     
     
         14 . The composition of  claim 1 , wherein the micellar nanoparticle composition exhibits a transfection efficiency of the at least one target cancer cell of between 10-fold and 100-fold greater than a micellar nanoparticle composition comprising a PEG polymer having an average molecular weight greater than 1 kDa. 
     
     
         15 . A method for preparing a polymeric micellar nanoparticle composition comprising a block or graft copolymer comprising at least one polycationic polymer and at least one polyethylene glycol (PEG) polymer having an average molecular weight less than 1 kDa; and at least one nucleic acid; wherein the graft or block copolymer and the at least one nucleic acid are complexed and condensed into a shaped micellar nanoparticle that is stable in biological media, the method comprising:
 (a) mixing a first solution comprising the block or graft copolymer together with a second solution comprising the at least one nucleic acid to form a third solution comprising the block or graft copolymer and the at least one nucleic acid; and   (b) allowing the block or graft copolymer and the at least one nucleic acid to self-assemble into the polymeric micellar nanoparticle.   
     
     
         16 . A transfection agent for transfecting a cell with at least one nucleic acid, the transfection agent comprising a polymeric micellar nanoparticle composition comprising a block or graft copolymer comprising at least one polycationic polymer and at least one polyethylene glycol (PEG) polymer having an average molecular weight less than 1 kDa; and at least one nucleic acid; wherein the graft or block copolymer and the at least one nucleic acid are complexed and condensed into a shaped micellar nanoparticle that is stable in biological media, wherein the polymeric micellar nanoparticle composition is prepared by the method of  claim 15 . 
     
     
         17 . The transfection agent of  claim 16 , wherein the transfection agent modulates expression of at least one gene in a cell, tissue, or subject. 
     
     
         18 . A method for targeting at least one metastatic cancer cell in a subject, the method comprising administering the transfection agent of  claim 16  to a subject, wherein the polymeric micellar nanoparticle composition comprises a ligand that binds to a tumor-specific antigen on the surface of the at least one metastatic cancer cell, and wherein the ligand binds to the tumor-specific antigen on the surface of the at least one metastatic cancer cell after administration of the transfection agent to the subject, thereby targeting the at least one metastatic cancer cell in the subject. 
     
     
         19 . The method of  claim 18 , wherein:
 (i) targeting the at least one metastatic cancer cell comprises treating a metastatic cancer in the subject; or   (ii) targeting the at least one metastatic cancer cell comprises detecting, diagnosing, and/or imaging a metastatic cancer in the subject.   
     
     
         20 . The method of  claim 18 , wherein the polymeric micellar nanoparticle composition further comprises a chemotherapeutic agent and/or at least one nucleic acid encoding a therapeutic gene that inhibits the growth, proliferation and/or survival of the at least one metastatic cancer cell. 
     
     
         21 . The method of  claim 18 , wherein the polymeric micellar nanoparticle composition further comprises an imaging agent and/or at least one nucleic acid encoding a reporter gene operably linked to a tumor-specific promoter. 
     
     
         22 . The method of  claim 21 , wherein the reporter gene is selected from the group consisting of a bioluminescent reporter gene, a fluorescent reporter gene, a CT reporter gene, an MRI reporter gene, a PET reporter gene, a SPECT reporter gene, and combinations thereof. 
     
     
         23 . The method of  claim 18 , further comprising imaging the subject after administering the transfection agent using an imaging modality selected from the group consisting of bioluminescent imaging, fluorescent imaging, CT, MRI, PET, SPECT, X-ray, and combinations thereof. 
     
     
         24 . A method for treating a disease or condition, the method comprising administering to a subject in need of treatment thereof, a polymeric micellar nanoparticle composition comprising a block or graft copolymer comprising at least one polycationic polymer and at least one polyethylene glycol (PEG) polymer having an average molecular weight less than 1 kDa; and at least one nucleic acid; wherein the graft or block copolymer and the at least one nucleic acid are complexed and condensed into a shaped micellar nanoparticle that is stable in biological media, or a pharmaceutical composition thereof, in an amount effective for treating the disease or condition. 
     
     
         25 . A method for preventing a disease or condition, the method comprising administering to a subject in need of prophylactic treatment thereof, a polymeric micellar nanoparticle composition comprising a block or graft copolymer comprising at least one polycationic polymer and at least one polyethylene glycol (PEG) polymer having an average molecular weight less than 1 kDa; and at least one nucleic acid; wherein the graft or block copolymer and the at least one nucleic acid are complexed and condensed into a shaped micellar nanoparticle that is stable in biological media, or a pharmaceutical composition thereof, in an amount effective for preventing the disease or condition.