Protein particles comprising disulfide crosslinkers and uses

Patent metadata

Field	Value
Publication number	US-9962423-B2
Application number	US-201414774764-A
Country	US
Kind code	B2
Filing date	Mar 11, 2014
Priority date	Mar 13, 2013
Publication date	May 8, 2018
Grant date	May 8, 2018

Abstract

This disclosure relates to particles comprising recombinant proteins, pharmaceutical composition comprising the particles, and therapeutic uses related thereto. In certain embodiments, the particles are made by the process of producing recombinant proteins and conjugating the recombinant proteins to form nanoparticles with a linking reagent comprising disulfide bonds. Typically the recombinant protein has a polypeptide of viral, fungal, or bacterial origin such as secreted effector proteins AvrA and YopJ. In certain embodiments, the disclosure relates to treating or preventing autoimmune diseases, cancer, or inflammatory diseases, or conditions such as inflammatory bowel disease (IBD).

First claim

The invention claimed is:

1. Particles comprising
a) recombinant AvrA proteins; and
b) linking groups comprising disulfide bonds,
wherein the linking groups conjugate the recombinant AvrA proteins to form the particles and wherein the size of the particles are between 50 nm and 200 nm in diameter.

2. The particles of claim 1 , wherein the recombinant AvrA is mAvrA(C186A).

3. The particles of claim 1 , wherein the recombinant AvrA proteins are fusion proteins comprising bovine serum albumin (BSA), green fluorescent protein (GFP), glutathione S-transferase (GST), polyhistidine, polylysine, or combinations thereof.

4. The particles of claim 1 , wherein the linking groups have the following formula,

wherein,
n is 1 to 100,
L forms amides to the recombinant AvrA proteins, and
X is at each occurrence individually and independently selected from O, NH, C═O, CH 2 , OCH 2 , CH 2 O, NHCH 2 , CH 2 NH, OCH 2 CH 2 , CH 2 CH 2 O, NHCH 2 CH 2 , or CH 2 CH 2 NH.

5. The particles of claim 4 , wherein X is (CH 2 )n wherein n is 1 to 4.

6. A pharmaceutical composition comprising the particles of claim 1 and a pharmaceutically acceptable excipient.

Assignees

• Univ Emory
• Georgia Tech Res Inst

Inventors

• Neish Andrew S
• Champion Julie A

Classifications

• A61K47/64

  Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent (peptidic linkers A61K47/65) · CPC title

• A61K47/6929

  the form being a nanoparticle, e.g. an immuno-nanoparticle · CPC title

• C07K2319/60

  containing spectroscopic/fluorescent detection, e.g. green fluorescent protein [GFP] · CPC title

• C07K14/255

  Salmonella (G) · CPC title

• C07K2319/21

  containing a His-tag · CPC title