Fabrication of protein-encapsulating microgels

Patent metadata

Field	Value
Publication number	US-12350374-B2
Application number	US-202117553507-A
Country	US
Kind code	B2
Filing date	Dec 16, 2021
Priority date	Dec 17, 2020
Publication date	Jul 8, 2025
Grant date	Jul 8, 2025

Abstract

The present invention provides methods for fabricating protein-encapsulating microgels using hydrocarbon-in-fluorocarbon emulsions. The non-aqueous emulsion-based microgel fabrication methods can be used for the encapsulation of a wide range of proteins and peptides, including antibodies and antibody-fusion proteins, for therapeutic use with ease of administration.

First claim

What is claimed is:

1. A method of producing microparticles comprising:
(a) combining a polyethylene glycol-amine polymer, a polyethylene glycol-N-hydroxysuccinimide polymer, a polyethylene glycol (PEG) polymer, and a powder including a vascular endothelial growth factor-Trap (VEGF-Trap) protein with dichloromethane to form a dispersed phase suspension;
(b) adding said dispersed phase suspension to a continuous phase solution, wherein said continuous phase solution includes perfluorotripentylamine and a triblock copolymer perfluoropolyether-b-polyethylene glycol-b-perfluoropolyether (PFPE-PEG-PFPE), to form a combined dispersed phase suspension and continuous phase solution;
(c) applying blending forces to said combined dispersed phase suspension and continuous phase solution to form a non-aqueous emulsion having multiple dichloromethane droplets including said polyethylene glycol-amine polymer, said polyethylene glycol-N-hydroxysuccinimide polymer and said powder including VEGF-Trap in the perfluorotripentylamine; and
(d) removing the dichloromethane and the perfluorotripentylamine from said non-aqueous emulsion to form isolated microparticles.

2. The method of claim 1 , wherein the molar ratio of said amine to said N-hydroxysuccinimide is between 1:1 and 1:2.

3. The method of claim 1 , wherein said polyethylene glycol-amine polymer and/or said polyethylene glycol-N-hydroxysuccinimide polymer is a 4-armed or an 8-armed compound.

4. The method of claim 1 , wherein said VEGF-Trap protein is a truncated form of VEGF-Trap protein.

5. The method of claim 1 , wherein said VEGF-Trap protein is aflibercept.

6. The method of claim 1 , wherein said isolated microparticles have a diameter between 1·m and 200·m as measured by imaging analysis.

7. The method of claim 1 , wherein said powder including a VEGF-Trap protein is micronized by spray-drying, electrospray drying, reversible precipitation, spray freezing, microtemplating, or a combination thereof.

8. The method of claim 1 , wherein said blending forces comprise homogenization, vortexing, sonication, cavitation, agitation, or a combination thereof.

9. The method of claim 1 , wherein said microparticles are sustained release microparticles.

10. The method of claim 1 , wherein a concentration of said powder including a VEGF-Trap protein in said dispersed phase suspension is between 1.0% and 30% w/v.

11. The method of claim 1 , wherein a total concentration of said polyethylene glycol-amine polymer and said polyethylene glycol-N-hydroxysuccinimide polymer in said dispersed phase suspension is between 5.0% and 35% w/v.

12. The method of claim 1 , wherein a concentration of said PFPE-PEG-PFPE in said continuous phase solution is between 0.1% and 5.0% w/v.

13. The method of claim 1 , further comprising suspending said isolated microparticles in a pharmaceutically acceptable formulation.

14. The method of claim 13 , wherein said formulation comprises pH buffered saline, an aqueous solution, or a non-aqueous solution.

15. The method of claim 1 , wherein said powder including a VEGF-Trap protein further comprises at least one excipient.

16. A microparticle made by the method of claim 1 .

17. The method of claim 1 , wherein a concentration of said PEG polymer in said dispersed phase suspension is between 5.0% and 35% w/v.

Assignees

• Regeneron Pharma

Inventors

• Chen Hunter
• Zhao Yiming

Classifications

• A61K47/10

  Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers · CPC title

• A61K38/179 Primary

  for growth factors; for growth regulators · CPC title

• A61K9/5089

  Processes · CPC title

• A61K9/06

  Ointments; Bases therefor; {Other semi-solid forms, e.g. creams, sticks, gels (composition of ointments, creams or gels A61K47/00)} · CPC title

• A61K38/00

  Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00) · CPC title