Single protein nanocapsules for protein delivery with long-term effect

Patent metadata

Field	Value
Publication number	US-9289504-B2
Application number	US-201013255229-A
Country	US
Kind code	B2
Filing date	Mar 9, 2010
Priority date	Mar 9, 2009
Publication date	Mar 22, 2016
Grant date	Mar 22, 2016

Abstract

A protein nanocapsule having a single-protein core and a thin polymer shell anchored covalently to the protein core.

First claim

We claim:

1. A single-protein nanocapsule comprising a single-protein core and an artificial membrane covalently anchored to and encapsulating said single-protein, said artificial membrane comprising:
(a) a polymer network comprising repeated units of at least two different monomer units which independently are positively charged, negatively charged, or uncharged, and hydrophobic or hydrophilic, wherein:
said polymer network is crosslinked by a crosslinker comprising two or more polymerizable groups capable of forming a crosslinked co-polymer between the single protein core and at least one monomer unit; and
said crosslinker is selected from the group consisting of N,N′-bis(acryloyl)cystamine, bis[2-(methacryloyloxy)ethyl]phosphate, and bisacryloylated polypeptide, and is disposed in the nanocapsule so as to be degradable in vivo;
(b) a surface charge at physiological pH which is positive charge, negative charge or no charge; and
(c) a modifiable moiety, wherein said single protein is more resistant to protease.

2. The single-protein capsule of claim 1 , wherein said monomer unit is selected from the group consisting of: acrylamide, 2-hydroxyethyl acrylate, N-isopropylacrylamide, sodium acrylate, 2-acryloylamido-2-methylpropanesulfonic sodium, allyl amine, N-(3-aminopropyl) methacrylamide hydrochloride, dimethylamino ethyl methacrylate, (3-acrylamidopropyl)trimethylammonium hydrochloride, methyl acrylate, and styrene.

3. The single-protein capsule of claim 1 , wherein said polymerizable group is selected from the group consisting of a vinyl group, an acryl group, and an alkylacryl group.

4. The single-protein capsule of claim 1 , wherein said artificial membrane is covalently anchored to said single-protein core at at least 3 locations on said single protein.

5. The single-protein capsule of claim 4 , wherein said locations are any of lysine, cysteine, threonine, glutamic acid, aspartic acid, serine, histidine, arginine, tyrosine, proline, and tryptophan.

6. The single-protein capsule of claim 1 , wherein said modifiable moiety comprises a reactive group modified by a surface modifying agent.

7. The single-protein capsule of claim 6 , wherein said surface modifying agent is selected from the group consisting of a small molecule, a polymer, a peptide, a polypeptide, a protein, an oligonucleotide, a polysaccharide, and an antibody.

8. The single-protein capsule of claim 6 , wherein said surface modifying agent is an agent that alters solubility of said single-protein capsule, emits light, or effects cell targeting, or cell penetration.

9. The single-protein capsule of claim 1 which is transducible into a cell.

10. A plurality of single-protein nanocapsules according to claim 1 , wherein the single proteins in said single-protein nanocapsules are same or different from each other.

11. A composition comprising a single-protein nanocapsule of claim 1 and a pharmaceutically acceptable carrier.

12. A composition comprising a plurality of single-protein nanocapsules according to claim 10 and a pharmaceutically acceptable carrier.

13. The single-protein capsule of claim 1 , wherein said surface feature is a charge or solubility of the nanocapsule.

14. A single-protein nanocapsule comprising a single-protein core and an artificial membrane covalently anchored to and encapsulating said single-protein, said artificial membrane comprising a polymer network comprising repeated units of at least two different monomer units which independently are positively charged, negatively charged, or uncharged, and hydrophobic or hydrophilic, wherein:
said polymer network is crosslinked by a bis[2-(methacryloyloxy)ethyl]phosphate crosslinker comprising two or more polymerizable groups capable of forming a crosslinked co-polymer between the single protein core and at least one monomer unit; and
said bis[2-(methacryloyloxy)ethyl]phosphate crosslinker is disposed in the nanocapsule so that the crosslinker:
(a) does not degrade at a first physiological pH; and
(b) degrades at a second physiological pH.

Assignees

• Lu Yunfeng
• Yan Ming
• Du Juanjuan
• Univ California

Inventors

• Lu Yunfeng
• Yan Ming
• Du Juanjuan

Classifications

• A61K47/58 Primary

  obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. poly[meth]acrylate, polyacrylamide, polystyrene, polyvinylpyrrolidone, polyvinylalcohol or polystyrene sulfonic acid resin · CPC title

• A61K9/0019

  Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner (non-active ingredients are additionally classified in A61K47/00) · CPC title

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• A61K9/5169 Primary

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