SELF-ASSEMBLING NANOPARTICLE DRUG DELIVERY SYSTEM
A self-assembling nanoparticle drug delivery system for the delivery of drugs including peptides, proteins, nucleic acids or synthetic chemical drugs is provided. The self-assembling nanoparticle drug delivery system described herein includes viral capsid proteins, such as Hepatitis B Virus core protein, encapsulating the drug, a lipid bi-layer envelope and targeting or facilitating molecules anchored in the lipid bilayer. A method for construction of the self-assembling nanoparticle drug delivery system is also provided.
1 .- 13 . (canceled)
14 . A method for constructing a self-assembling nanoparticle drug delivery system comprising:
mixing a drug with a hepatitis B virus (HBV) core protein to form a cage solution;
encapsulating said drug in a capsid comprised of HBV core protein by raising the ionic strength of said cage solution to form a nanoparticle; and
purifying said nanoparticles.
15 . The method of claim 14 wherein said drug is selected from the group consisting of peptides, proteins, nucleic acids and small molecule synthetic chemical drugs.
16 . The method of claim 30 , wherein said HBV core protein includes a protease recognition site replacing amino acids 79 and 80 of SEQ ID NO: 1 or SEQ ID NO: 2.
17 . The method of claim 16 wherein said protease recognition site is a thrombin recognition site or a factor Xa recognition site.
18 . The method of claim 30 , wherein said HBV core protein is mutated such that at least one amino acid of SEQ ID NO: 1 or SEQ ID NO: 2 selected from the group consisting of phenylalanine 23, aspartic acid 29, threonine 33, leucine 37, valine 120, valine 124, arginine 127, tyrosine 132, glutamic acid 77, aspartic acid 78 and alanine 80 is changed to a cysteine.
19 . The method of claim 14 further comprising adding an envelopment guiding protein or peptide after said encapsulating step.
20 . The method of claim 19 wherein said envelopment guiding protein is Hepatitis B Virus S-protein or the transmembrane engineered peptide of SEQ ID NO:5.
21 . The method of claim 25 , wherein said phospholipid is selected from the group consisting of: phosphatidyl ethanolamine, phosphatidyl glycerol and hydrogenated soy phosphatidyl choline.
22 . The method of claim 27 , said protein transduction domain comprises the Human Immunodeficiency Virus transactivator or polyarginine.
23 . The method of claim 25 , further comprising inserting targeting antibodies into said lipid envelope.
24 . (canceled)
25 . The method of claim 14 , further comprising adding phospholipid to said cage solution to form a lipid envelope surrounding the nanoparticle.
26 . The method of claim 14 , further comprising adding a cholesterol-tagged polyethylene glycol to said cage solution.
27 . The method of claim 25 , further comprising adding a cholesterol-tagged protein transduction domain to said cage solution.
28 . The method of claim 14 , wherein the nanoparticle is purified by centrifugation.
29 . The method of claim 14 , wherein the nanoparticle is purified by size exclusion chromatography.
30 . The method of claim 14 , wherein the HBV core protein comprises the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 2.
31 . The method of claim 14 , wherein the HBV core protein is a modified HBV core protein.
32 . The method of claim 30 , wherein the glutamic acid at amino acid 77 of SEQ ID NO: 1 or SEQ ID NO: 2 is changed to a cysteine.
33 . The method of claim 25 , wherein the lipid envelope is covalently attached to said nanoparticle by a phosphatidyl-ethanolamine maleimide to the cysteine acid 77 of SEQ ID NO: 1 or SEQ ID NO: 2.