Lipid Nanoparticle Compositions and Methods of Making and Methods of Using the Same

Abstract

Lipid nanoparticle formulations, methods of making, and methods of using same are disclosed.

What is claimed is: 
     
         1 . A lipid nanoparticle comprising a combination of tertiary and quaternary amine-based cationic lipids, except not, wherein said combination consists of the tertiary amine-cationic lipid present at about 40.0 molar percent; wherein the tertiary amine-cationic lipid comprises N-[1-(2, 3-dioleyloyx) propyl]-N—N—N-dimethyl ammonium chloride (DODMA), and the concentration of the quaternary amine-cationic lipid present at about 5.0 molar percent. 
     
     
         2 . The lipid nanoparticle of  claim 1 , wherein the tertiary amine-cationic lipids are chosen from DODAP, DODMA, DC-CHOL, N,N-dimethylhexadecylamine, or combinations thereof. 
     
     
         3 . The lipid nanoparticle of  claim 1 , wherein the quaternary amine-cationic lipids are selected from DOTAP, DOTMA, DDAB, or combinations thereof. 
     
     
         4 . The lipid nanoparticle of  claim 1 , wherein the concentration of the tertiary amine-cationic lipids is below 60.0 molar percent of the total lipid content. 
     
     
         5 . The lipid nanoparticle of  claim 1 , wherein the concentration of quaternary amine-cationic lipids is below 20.0 molar percent of the total lipid content. 
     
     
         6 . The lipid nanoparticle of  claim 1 , wherein the nanoparticle encapsulates molecules selected from nucleic acids, proteins, polysaccharides, lipids, radioactive substances, therapeutic agents, prodrugs, nutritional supplements, biomarkers, or combinations thereof. 
     
     
         7 . The lipid nanoparticle of  claim 6 , wherein the encapsulated molecules comprise a nucleic acid selected from plasmid DNAs, antisense oligonucleotides, miRs, anti-miRs, shRNAs, siRNAs, or combinations thereof. 
     
     
         8 . The lipid nanoparticle of  claim 1 , further comprising a cationic polymer. 
     
     
         9 . The lipid nanoparticle of  claim 8 , wherein the cationic polymer is selected from the group consisting of: spermine, dispermine, trispermine, tetraspermine, oligospermine, thermine, spermidine, dispermidine, trispermidine, oligospermidine, putrescine, polylysine, polyarginine, a polyethylenimine of branched or linear type, and polyallylamine. 
     
     
         10 . The lipid nanoparticle of  claim 1 , further comprising a fusogenic peptide. 
     
     
         11 . The lipid nanoparticle of  claim 6 , wherein the encapsulation rate of therapeutic agents or nucleotides is 20% or higher. 
     
     
         12 . The lipid nanoparticle of  claim 1 , wherein the lipid nanoparticle has a diameter under 300 nm. 
     
     
         13 . The lipid nanoparticle of  claim 1 , wherein the lipid nanoparticle comprises the lipids DODMA and DOTMA in a molar ratio selected from 45:0, 5:40, 15:30, 22.5:22.5, 30:15, or 40:5. 
     
     
         14 . The lipid nanoparticle of  claim 1 , wherein the lipid nanoparticle comprises the lipids DMHDA and DOTAP in a molar ratio selected from 90:10, 70:30, 50:50, 30:70, or 10:90. 
     
     
         15 . A lipid nanoparticle having a diameter of less than 300 nm and comprising a peptide. 
     
     
         16 . The lipid nanoparticle of  claim 15 , wherein the peptide is selected from gramicidin A, B, C, D, or S; JTS-1; proteinase K (PrK); trichorovin-Xlla; rabies virus glycoprotein; interleukin-1 β; HIV-Tat; herpes simplex virus VP22 protein; and combinations thereof. 
     
     
         17 . The lipid nanoparticle of  claim 15 , wherein the peptide comprises an antibiotic. 
     
     
         18 . The lipid nanoparticle of  claim 17 , wherein the antibiotic is selected from gramicidin A, B, C, D, or S. 
     
     
         19 . The lipid nanoparticle of  claim 15 , wherein the peptide consists essentially of a lipidated JTS-1 fusogenic peptide. 
     
     
         20 . The lipid nanoparticle of  claim 19 , wherein the lapidated JTS-1 fusogenic peptide is present at about 0 to about 30 molar percent of the total formulation. 
     
     
         21 . The lipid nanoparticle of  claim 15 , further comprising proteinase K. 
     
     
         22 . The lipid nanoparticle of  claim 15 , wherein the proteinase K is present at about 0 to about 30 molar percent of the total formulation. 
     
     
         23 . The lipid nanoparticle of  claim 15 , wherein the lipid nanoparticle encapsulates molecules selected from nucleic acids, proteins, polysaccharides, lipids, radioactive substances, therapeutic agents, prodrugs, nutritional supplements, biomarkers, or combinations thereof. 
     
     
         24 . The lipid nanoparticle of  claim 23 , wherein the encapsulated molecules comprise a nucleic acid selected from plasmid DNAs, antisense oligonucleotides, miRs, anti-miRs, shRNAs, siRNAs, or combinations thereof. 
     
     
         25 . A lipid nanoparticle comprising a DNase- or RNase-degrading agent. 
     
     
         26 . The lipid nanoparticle of  claim 25 , wherein the DNase- or RNase-degrading agent consists essentially of proteinase K. 
     
     
         27 . The lipid nanoparticle of  claim 25 , wherein the nanoparticle encapsulates molecules selected from nucleic acids, proteins, polysaccharides, lipids, radioactive substances, therapeutic agents, prodrugs, nutritional supplements, biomarkers, or combinations thereof. 
     
     
         28 . The lipid nanoparticle of  claim 27 , wherein the encapsulated molecules comprise an oligonucleotide selected from pDNAs, antisense oligonucleotides, miRs, anti-miRs, shRNAs, siRNAs, or combinations thereof. 
     
     
         29 . The lipid nanoparticle of  claim 25 , wherein the lipid nanoparticle has a diameter under 300 nm. 
     
     
         30 . A lipid nanoparticle having a diameter of less than 300 nm and comprising a combination of two or more of:
 a mixture of tertiary and quaternary amine-cationic head groups;   an antibiotic; or   a DNase or RNase-degrading agent.   
     
     
         31 . The lipid nanoparticle of  claim 30 , wherein the RNase or RNase-degrading agent consists essentially of proteinase K. 
     
     
         32 . The lipid nanoparticle of  claim 30 , wherein the antibiotic comprises gramicidin A, B, C, D, or S. 
     
     
         33 . A lipid nanoparticle of  claim 1 , further comprising a polyethyleneglycol-lipid conjugate. 
     
     
         34 . The lipid nanoparticle of  claim 33 , wherein the polyethyleneglycol-lipid conjugate selected from polysorbate 80, TPGS, mPEG-DSPE, PEG-DMG, DPPE-PEG or mPEG-DMPE. 
     
     
         35 . The lipid nanoparticle of  claim 33 , wherein the polyethyleneglycol-lipid is present at a concentration less than about 10.0 molar percent. 
     
     
         36 . A lipid nanoparticle of  claim 33 , further comprising N,N-dimethylhexadecylamine. 
     
     
         37 . The lipid nanoparticle of  claim 36 , wherein the N,N-dimethylhexadecylamine is present at a concentration of less than about 60.0 molar percent of the formulation. 
     
     
         38 . A lipid nanoparticle of  claim 36 , further comprising a ligand capable of binding to a target cell or a target molecule. 
     
     
         39 . The lipid nanoparticle of  claim 38 , wherein the ligand is an antibody or an antibody fragment. 
     
     
         40 . The lipid nanoparticle of  claim 38 , wherein the ligand is selected from cRGD, galatose-containing moieties, transferrin, folate, low density lipoprotein, or epidermal growth factors. 
     
     
         41 . A pharmaceutical composition comprising a lipid nanoparticle of  claim 1 , and a pharmaceutically acceptable excipient. 
     
     
         42 . The pharmaceutical composition of  claim 41 , wherein the pharmaceutical composition is administered perorally, intravenously, intraperitoneally, subcutaneously, or transdermally. 
     
     
         43 . The pharmaceutical composition of  claim 41 , wherein the pharmaceutical composition is prepared as an orally administered tablet. 
     
     
         44 . The pharmaceutical composition of  claim 41 , wherein the pharmaceutical composition is prepared as a sterile solution. 
     
     
         45 . The pharmaceutical composition of  claim 41 , wherein the pharmaceutical composition is prepared as a sterile suspension. 
     
     
         46 . The pharmaceutical composition of  claim 41 , wherein the pharmaceutical composition is prepared as a lyophilized powder. 
     
     
         47 . The pharmaceutical composition of  claim 41 , wherein the pharmaceutical composition is prepared as a suppository. 
     
     
         48 . A method of diagnosing or treating a cancer or infectious disease, the method comprising administering an effective amount of a pharmaceutical composition of  claim 41  to a patient in need thereof. 
     
     
         49 . The lipid nanoparticle of  claim 1 , wherein the lipid nanoparticle encapsulates at least one molecule selected from nucleic acids, chemotherapeutic agents, and combinations thereof. 
     
     
         50 . The lipid nanoparticle of  claim 49 , wherein the encapsulated molecule comprises a nucleic acid selected from plasmid DNAs, antisense oligonucleotides, miRs, anti-miRs, shRNAs, siRNAs, and combinations thereof. 
     
     
         51 . The lipid nanoparticle of  claim 49 , wherein the encapsulated molecule comprises a therapeutic agent selected from: antineoplastic agents, anti-infective agents, local anesthetics, anti-allergics, antianemics, angiogenesis, inhibitors, beta-adrenergic blockers, calcium channel antagonists, anti-hypertensive agents, anti-depressants, anti-convulsants, anti-bacterial, anti-fungal, anti-viral, anti-rheumatics, anthelminithics, antiparasitic agents, corticosteroids, hormones, hormone antagonists, immunomodulators, neurotransmitter antagonists, anti-diabetic agents, anti-epileptics, anti-hemmorhagics, anti-hypertonics, antiglaucoma agents, immunomodulatory cytokines, sedatives, chemokines, vitamins, toxins, narcotics, imaging agents, and combinations thereof. 
     
     
         52 . The lipid nanoparticle of  claim 49 , wherein the encapsulated molecule comprises a nucleic acid therapeutic agent. 
     
     
         53 . The lipid nanoparticle of  claim 52 , wherein the nucleic acid therapeutic agent is selected from: pDNA, siRNA, miRNA, anti-miRNA, ASO, and combinations thereof. 
     
     
         54 . The lipid nanoparticle of  claim 52 , wherein the nucleic acid therapeutic agent is stabilized by modifications to substituent NA base units and/or by modifying the ribose 2′ position or substituting phosphodiester linkers. 
     
     
         55 . The lipid nanoparticle of  claim 1 , wherein the lipid nanoparticle has a diameter under about 300 nm. 
     
     
         56 . The lipid nanoparticle of  claim 1 , wherein the lipid nanoparticle has a diameter under about 200 nm. 
     
     
         57 . The lipid nanoparticle of  claim 1 , wherein the polymer is bound to an external surface of the lipid via direct connection or via a linker. 
     
     
         58 . The lipid nanoparticle of  claim 16 , wherein the lipid nanoparticle has an encapsulation efficiency of the molecule of at least about 40%. 
     
     
         59 . The lipid nanoparticle of  claim 1 , further including a polyethylene glycol-conjugated lipid. 
     
     
         60 . The lipid nanoparticle of  claim 59 , wherein the polyethylene glycol-conjugated lipid comprises one or more of: polysorbate 80, TPGS, DPPE-PEG, mPEG-DMPE, and mPEG-DSPE. 
     
     
         61 . The lipid nanoparticle of  claim 27 , wherein the polyethylene glycol-conjugated lipid is present at a concentration less than about 15.0 molar percent. 
     
     
         62 . The lipid nanoparticle of  claim 1 , further comprising a ligand capable of binding to a target cell or a target molecule. 
     
     
         63 . The lipid nanoparticle of  claim 62 , wherein the ligand is an antibody or an antibody fragment. 
     
     
         64 . The lipid nanoparticle of  claim 62 , wherein the ligand is selected from cRGD, galatose-containing moieties, transferrin, folate, low density lipoprotein, or epidermal growth factors. 
     
     
         65 . A pharmaceutical composition comprising the lipid nanoparticle of  claim 1 , and a pharmaceutically acceptable excipient. 
     
     
         66 . The pharmaceutical composition of  claim 65 , wherein the pharmaceutical composition is administered perorally, intravenously, intraperitoneally, subcutaneously, or transdermally. 
     
     
         67 . The pharmaceutical composition of  claim 65 , wherein the pharmaceutical composition is prepared as an orally administered tablet, an inhalant, or a suppository. 
     
     
         68 . The pharmaceutical composition of  claim 65 , wherein the pharmaceutical composition is prepared as a sterile solution, a sterile suspension, or a lyophilized powder. 
     
     
         69 . A lipid nanoparticle comprising, an anti-miR-221 and SPLN-G20, wherein SPLN-G20 comprises: DMHDA, DOTAP, GRAM, DOPE and TPGS at a molar ratio of 40:5:20:30:5. 
     
     
         70 . The lipid nanoparticle of  claim 69 , wherein the anti-miR-221 has the sequence: 5′-g s a s aacccagcagacaaugu s a s g s c s u-Chol-3′, SEQ ID NO. 1. 
     
     
         71 . The lipid nanoparticle of  claim 70 , wherein the sequence includes 2′-O-Methyl-modified oligonucleotides (lower case letters) and phosphorothioate linkages (s subscript) to increase nuclease stability of the oligonucleotides. 
     
     
         72 . A composition comprising anti-miR-221 combined a lipid nanoparticle. 
     
     
         73 . The composition of  claim 72 , wherein the lipid nanoparticle comprises SPLN-G20v1 and SPLN-G20v2, wherein SPLN-G20 version 1 (SPLN-G20v1) comprises DMHDA, DOTAP, GRAM, DOPE and TPGS at a molar ratio of 40:5:20:30:5, and wherein SPLN-G20 (SPLN-G20v2) comprises DODAP, DOTAP, GRAM, Soy PC (SPC) and TPGS at a molar ratio of 40:5:20:30:5. 
     
     
         74 . The composition of  claim 73 , wherein the lipid nanoparticle comprises SPLN-G20v1 and SPLN-G20v2 at an N:P of 15:1. 
     
     
         75 . A method of treating a subject having, or suspected of having, breast cancer, comprising administering an effective amount of the lipid nanoparticle of  claim 69 . 
     
     
         76 . A method of providing anti-oncogenic effect in a subject suffered from breast cancer, in which miR-221 expression level is higher in cancer cells of case subject relative to a control subject, comprising administering to the subject an effective amount of a composition of  claim 69 . 
     
     
         77 . The method of  claim 76 , wherein the lipid nanoparticle down-regulates miR-221 expression. 
     
     
         78 . The method of  claim 76 , wherein the breast cancer comprises triple negative breast cancer. 
     
     
         79 . The method of  claim 78 , wherein the subject is a human. 
     
     
         80 . A composition comprising anti-miR-155 combined with a lipid nanoparticle. 
     
     
         81 . The composition of  claim 80 , wherein the lipid nanoparticle comprises Lac-GLN, wherein Lac-GLN comprises a lipophilic asialoglycoprotein receptor (ASGR) targeting ligand composed of lactobionic acid (LA), bearing a galactose moiety, and linked to a phospholipid. 
     
     
         82 . The composition of  claim 80 , further including gramicidin A incorporated into the lipid nanoparticle. 
     
     
         83 . The composition of  claim 82 , comprising DODAP, Lac-DOPE, DOPE, DMG-PEG and gramicidin A at a molar ratio of 50:10:28:2:10. 
     
     
         84 . The composition of  claim 80 , wherein the anti-miR-155 has the sequence: 5′-A*C*CCCUAUCACGAUUAGCAUU*A*A-3′, SEQ ID NO. 6. 
     
     
         85 . The composition of  claim 84 , wherein the sequence contains phosphorothioate linkages (*) and 2′-O-Methyl. 
     
     
         86 . A method of treating a subject having, or suspected of having liver cancer, comprising administering an effective amount of the lipid nanoparticle of  claim 80 . 
     
     
         87 . A method of providing anti-oncogenic effect in a subject suffered from liver cancer, in which miR-155 expression level is higher in cancer cells of case subject relative to a control subject, comprising administering to the subject an effective amount of the lipid nanoparticle of  claim 80 . 
     
     
         88 . The method of  claim 87 , wherein the lipid nanoparticle down-regulates C/EBPβ and FOXP3 genes. 
     
     
         89 . The method of  claim 86 , wherein the subject is a human.