Lipid nanoparticle compositions and methods of making and methods of using the same

1. A nanoparticle comprising:

a combination of tertiary and quaternary amine-based cationic lipids,

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;

and at least one single stranded oligonucleotide encapsulated within the lipid combination.

2. The nanoparticle of claim 1 , wherein the tertiary amine-cationic lipid further comprises one or more of: lipids are chosen from 1,2-dioleoyl-3-dimethylammonium-propane (DODAP), 3β-[N—(N′,N′-dimethylaminoethane)-carbamoyl]cholesterol hydrochloride (DC-CHOL), N,N-dimethylhexadecylamine (DMHDA), and combinations thereof.

3. The nanoparticle of claim 1 , wherein the quaternary amine-cationic lipids are selected from 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP), N-[1-(2, 3-dioleyloyx) propyl]-N—N—N-trimethyl ammonium chloride (DOTMA), dimethyldioctadecylammonium bromide (DDAB), or combinations thereof.

4. The nanoparticle of claim 1 , wherein the encapsulated single stranded oligonucleotide comprises: antisense oligonucleotides, miRs, anti-miRs, or combinations thereof.

5. The nanoparticle of claim 1 , further comprising a cationic polymer.

6. The nanoparticle of claim 5 , wherein the cationic polymer is selected from the group consisting of: spermine, dispermine, dispermine, tetraspermine, oligospermine, thermine, spermidine, dispermidine, trispermidine, oligospermidine, putrescine, polylysine, polyarginine, a polyethylenimine of branched or linear type, and polyallylamine.

7. The nanoparticle of claim 1 , further comprising a fusogenic peptide covalently bonded to, or conjugated to the lipids.

8. The nanoparticle of claim 1 , wherein the encapsulation rate is 20% or higher.

9. The nanoparticle of claim 1 , wherein the nanoparticle has a diameter under 300 nm.

10. A nanoparticle having a diameter of less than 300 nm; comprising: the nanoparticle of claim 1 ; and

at least one peptide mixed with, covalently attached or conjugated to the lipids.

11. The nanoparticle of claim 10 , 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.

12. The nanoparticle of claim 10 , wherein the peptide comprises an antibiotic.

13. The nanoparticle of claim 12 , wherein the antibiotic is selected from gramicidin A, B, C, D, or S.

14. The nanoparticle of claim 10 , wherein the peptide consists essentially of a lipidated JTS-1 fusogenic peptide.

15. The nanoparticle of claim 14 , wherein the lipidated JTS-1 fusogenic peptide is present at about 0 to about 30 molar percent of the total formulation.

16. The nanoparticle of claim 10 , further comprising proteinase K.

17. The nanoparticle of claim 16 , wherein the proteinase K is present at about 0 to about 30 molar percent of the total formulation.

18. The nanoparticle of claim 10 , wherein the encapsulated single stranded oligonucleotide comprises: antisense oligonucleotides, miRs, anti-miRs, or combinations thereof.

19. The nanoparticle of claim 1 , further comprising a hydrophilic polymer covalently bonded to the lipids, or conjugated to the lipids.

20. The nanoparticle of claim 19 , wherein the hydrophilic polymer is selected from polysorbate 80, D-alpha-tocopheryl polyethylene glycol 1000 succinate (TPGS), mPEG-1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N (DSPE), methoxypolyethylene glycol (PEG-DMG), dipalmitoylphosphatidlyethanolamine-PEG2000 (DPPE-PEG) or mPEG dimyristoylphosphatidylethanolamine (DMPE).

21. The nanoparticle of claim 20 , wherein the hydrophilic polymer is present at a concentration less than about 10.0 molar percent.

22. The nanoparticle of claim 1 , further comprising a ligand capable of binding to a target cell or a target molecule.

23. The nanoparticle of claim 22 , wherein the ligand is an antibody or an antibody fragment.

24. The nanoparticle of claim 22 , wherein the ligand is selected from cRGD, galatose-containing moieties, transferrin, folate, low density lipoprotein, or epidermal growth factors.

25. The nanoparticle of claim 1 , wherein the nanoparticle further encapsulates an at least one additional 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.

26. The nanoparticle of claim 25 , wherein the encapsulated therapeutic agent comprises a nucleic acid therapeutic agent.

27. The nanoparticle of claim 26 , wherein the nucleic acid therapeutic agent is selected from: pDNA, miRNA, anti-miRNA, antisense oligonucleotide (ASO), and combinations thereof.

28. The nanoparticle of claim 26 , wherein the nucleic acid therapeutic agent is stabilized by modifications to substituent nucleic acid base units and/or by modifying the ribose 2′ position or substituting phosphodiester linkers.

29. The nanoparticle of claim 1 , wherein the lipid nanoparticle has a diameter under about 200 nm.

30. The nanoparticle of claim 19 , wherein the hydrophilic polymer is bound to an external surface of the nanoparticle via direct connection or via a linker.

31. The nanoparticle of claim 11 , wherein the nanoparticle has an encapsulation efficiency of the molecule of at least about 40%.

32. A pharmaceutical composition comprising the nanoparticle of claim 1 , and a pharmaceutically acceptable excipient.

33. The pharmaceutical composition of claim 32 , wherein the pharmaceutical composition is a tablet, an inhalant, or a suppository.

34. The pharmaceutical composition of claim 32 , wherein the pharmaceutical composition is a sterile solution, a sterile suspension, or a lyophilized powder.

35. A composition comprising anti-miR-221 combined nanoparticle of claim 1 .

36. A composition comprising anti-miR-155 combined with the nanoparticle of claim 1 .

37. The composition of claim 36 , 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.

38. The composition of claim 36 , further including gramicidin A incorporated into the lipid nanoparticle.

39. The composition of claim 36 , wherein the anti-miR-155 has the sequence: 5′-A*C*CCCUAUCACGAUUAGCAUU*A*A-3′, SEQ ID NO. 6.

40. The composition of claim 39 , wherein the sequence contains phosphorothioate linkages (*) and 2′-O-Methyl.