Protein vesicles and methods of making and using thereof

Patent metadata

Field	Value
Publication number	US-2016158154-A1
Application number	US-201514961616-A
Country	US
Kind code	A1
Filing date	Dec 7, 2015
Priority date	Dec 5, 2014
Publication date	Jun 9, 2016
Grant date	—

Abstract

Hollow, protein vesicles are provided. The protein vesicles can be loaded with desired cargo to be delivered to a subject, for example a human. One embodiment provides a protein vesicle made of a protein membrane surrounding a hollow core. The protein membrane includes a first and a second modular protein amphiphile. The first modular protein amphiphile includes a hydrophobic block and a first hydrophilic protein binding block. The second modular protein amphiphile includes a variable polypeptide block and a second hydrophilic protein binding block that binds to the first hydrophilic protein binding block. The first and second protein amphiphiles self-assemble to form the hollow, protein vesicle.

First claim

We claim:

1 . A hollow, protein vesicle comprising:
a protein membrane surrounding a hollow core, wherein the protein membrane comprises a first and a second modular protein amphiphile, wherein the first modular protein amphiphile comprises a hydrophobic block and a first hydrophilic protein binding block and the second modular protein amphiphile comprises a variable polypeptide block and a second hydrophilic protein binding block that binds to the first hydrophilic protein binding block; and wherein the first and second protein amphiphiles self-assemble to form the hollow, protein vesicle.

2 . The hollow, protein vesicle of claim 1 , wherein the hydrophobic block of the first modular protein amphiphile comprises elastin-like polypeptide.

3 . The hollow, protein vesicle of claim 2 , wherein the first and second hydrophilic protein binding blocks comprise a leucine zipper motif.

4 . The hollow, protein vesicle of claim 3 , wherein the first hydrophilic protein binding block comprises an arginine-rich leucine zipper motif and the second hydrophilic protein binding block comprises a glutamic acid-rich leucine zipper motif.

5 . The hollow, protein vesicle of claim 1 , wherein the second hydrophilic block comprises a catalytic domain of an enzyme, a targeting moiety, a fluorescent polypeptide, a Human Leukocyte Antigen polypeptide, a T cell receptor polypeptide, a detectable label, a globular polypeptide, an immunostimulatory polypeptide, or combinations thereof.

6 . The hollow, protein vesicle of claim 5 , wherein the enzyme is selected from the group consisting of a dehydrogenase, a reductase, a protease, a synthetase.

7 . The hollow, protein vesicle of claim 5 , wherein the targeting moiety is selected from the group consisting of an antibody, aptamer, ligand receptor, and fusion protein.

8 . The hollow, protein vesicle of claim 7 , wherein the antibody is selected from the group consisting of a monoclonal antibody, a polyclonal antibody, a diabody, a single chain antibody, and antigen binding fragments thereof.

9 . A method for making hollow, protein vesicles comprising:
combining a plurality of first and second modular protein amphiphiles in a solution on ice; adjusting the salt concentration to promote or allow inverse phase transition of the protein amphiphiles; and incubating the salt-adjusted solution at room temperature to form hollow, protein vesicles.

10 . The method of claim 9 , wherein the first modular protein amphiphile comprises a hydrophobic block and a first hydrophilic protein binding block and the second modular protein amphiphile comprises a variable polypeptide block and a second hydrophilic protein binding block that binds to the first hydrophilic protein binding block.

11 . The method of claim 10 , wherein the hydrophobic block of the first modular protein amphiphile comprises elastin-like polypeptide.

12 . The method of claim 11 , wherein the first and second hydrophilic protein binding blocks comprise a leucine zipper motif.

13 . The method of claim 12 , wherein the first hydrophilic protein binding block comprises an arginine-rich leucine zipper motif and the second hydrophilic protein binding block comprises a glutamic acid-rich leucine zipper motif.

14 . A hollow, protein vesicle produced according to the method of claim 9 .

15 . A loaded, protein vesicle comprising:
a protein membrane surrounding a core, wherein the protein membrane comprises a first and a second modular protein amphiphile, wherein the first modular protein amphiphile comprises a hydrophobic block and a first hydrophilic protein binding block and the second modular protein amphiphile comprises a variable polypeptide block and a second hydrophilic protein binding block that binds to the first hydrophilic protein binding block; and wherein the first and second protein amphiphiles self-assemble around cargo to form the protein vesicle with the cargo loaded in the core of the protein vesicle.

16 . The loaded, protein vesicle of claim 15 , wherein the hydrophobic block of the first modular protein amphiphile comprises elastin-like polypeptide.

17 . The loaded, protein vesicle of claim 16 , wherein the first and second hydrophilic protein binding blocks comprise a leucine zipper motif.

18 . The loaded, protein vesicle of claim 17 , wherein the first hydrophilic protein binding block comprises an arginine-rich leucine zipper motif and the second hydrophilic protein binding block comprises a glutamic acid-rich leucine zipper motif.

19 . The loaded, protein vesicle of claim 16 , wherein the cargo comprises a drug, a protein, a contrast agent, nanoparticles, a fluorophore, a radiolabel, a therapeutic compound, an antioxidant, a growth factor, a cytokine, a chemoattractant, a nucleic acid or a combination thereof.

20 . The loaded, protein vesicle of claim 19 , wherein the drug is selected from the group consisting of a cytocidal compound, a chemotherapy agent, a cellular metabolism blocker, a cell cycle inhibitor, or a combination thereof.

21 . The loaded, protein vesicle of claim 19 , wherein the nucleic acid encodes a functional protein.

22 . The nucleic acid of claim 19 , wherein the nucleic acid is selected from the group consisting of double stranded DNA, RNA, siRNA, microRNA, single stranded DNA, and anti-sense DNA.

Assignees

• Georgia Tech Res Inst

Inventors

• Champion Julie A
• Park Won Min

Classifications

• A61K9/1271 Primary

  Non-conventional liposomes, e.g. PEGylated liposomes or liposomes coated or grafted with polymers (liposomes as conjugates {A61K47/6911}) · CPC title

• A61K9/1277

  Preparation processes; Proliposomes · CPC title

• A61K49/0002

  General or multifunctional contrast agents, e.g. chelated agents · CPC title

• A61K9/5169 Primary

  Proteins, e.g. albumin, gelatin · CPC title