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 . A self-assembling nanoparticle drug delivery system comprising:
a capsid comprised of viral capsid proteins;
a drug captured in said capsid; and
a lipid bi-layer enveloping said capsid.
2 . The self-assembling nanoparticle drug delivery system of claim 1 wherein said viral capsid protein is Hepatitis B Virus (HBV) core protein.
3 . The self-assembling nanoparticle drug delivery system of claim 1 wherein said HBV core protein has the amino acid sequence of SEQ ID NO. 1 or SEQ ID NO. 2.
4 . The self-assembling nanoparticle drug delivery system of claim 1 wherein said viral capsid protein is mutated.
5 . The self-assembling nanoparticle drug delivery system of claim 4 wherein said viral capsid protein includes a protease recognition site replacing amino acids 79 and 80 of said HBV core protein.
6 . The self-assembling nanoparticle drug delivery system of claim 5 wherein said protease recognition site is a thrombin recognition site or a factor Xa recognition site.
7 . The self-assembling nanoparticle drug delivery system of claim 4 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 and tyrosine 132 is changed to a cysteine.
8 . The self-assembling nanoparticle drug delivery system of claim 1 wherein said drug is selected from the group consisting of peptides, proteins, nucleic acids and small molecule synthetic chemical drugs.
9 . The self-assembling nanoparticle drug delivery system of claim 1 wherein said lipid bi-layer is comprised of phospholipids.
10 . The self-assembling nanoparticle drug delivery system of claim 9 wherein said phospholipid is phosphotidyl ethanolamine.
11 . The self-assembling nanoparticle drug delivery system of claim 1 further comprising either or both of cholesterol-tagged polyethylene glycol and cholesterol-tagged protein transduction domains.
12 . The self-assembling nanoparticle drug delivery system of claim 11 wherein said protein transduction domains comprises the Human Immunodeficiency Virus transactivator of transcription or poly-arginine.
13 . The self-assembling nanoparticle drug delivery system of claim 1 further comprising an antibody targeting molecule.
14 . A method for constructing a self-assembling nanoparticle drug delivery system comprising:
mixing a drug with HBV core protein to form a cage solution;
encapsulating said drug in the core protein cage by raising the ionic strength of said cage solution;
adding phospholipids to said cage solution;
adding cholesterol-tagged polyethylene glycol to said cage solution;
adding cholesterol-tagged protein transduction domain to said cage solution;
purifying said nanoparticles by centrifugation or size exclusion chromatography.
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 14 wherein said HBV core protein includes a protease recognition site replacing amino acids 79 and 80 of said HBV core protein.
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 14 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 and tyrosine 132 is changed to a cysteine.
19 . The method of claim 14 further comprising the step of 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 14 wherein said phospholipid is phosphatidyl ethanolamine.
22 . The method of claim 14 wherein said protein transduction domain comprises the Human Immunodeficiency Virus transactivator of transcription or poly-arginine.
23 . The method of claim 14 further comprising the step of inserting targeting antibodies into said lipid bi-layer.
24 . A method of treating disease with a self-assembling nanoparticle drug delivery system comprising delivering said nanoparticles across a mucosal surface.