Methods of preparing polyelectrolyte complex nanoparticles

Patent metadata

Field	Value
Publication number	US-2017042829-A1
Application number	US-201615221189-A
Country	US
Kind code	A1
Filing date	Jul 27, 2016
Priority date	Aug 13, 2015
Publication date	Feb 16, 2017
Grant date	—

Abstract

The presently disclosed subject matter provides methods for continuously generating uniform polyelectrolyte complex (PEC) nanoparticles comprising: flowing a first stream comprising one or more water-soluble polycationic polymers at a first variable flow rate into a confined chamber; flowing a second stream comprising one or more water-soluble polyanionic polymers at a second variable flow rate into the confined chamber; and impinging the first stream and the second stream in the confined chamber until the Reynolds number is from about 1,000 to about 20,000, thereby causing the one or more water-soluble polycationic polymers and the one or more water-soluble polyanionic polymers to undergo a polyelectrolyte complexation process that continuously generates PEC nanoparticles. Compositions produced from the presently disclosed methods and a device for producing the compositions are also disclosed.

First claim

That which is claimed:

1 . A flash nanocomplexation (FNC) method of continuously generating uniform polyelectrolyte complex (PEC) nanoparticles, the method comprising:
(a) flowing a first stream comprising one or more water-soluble polycationic polymers at a first variable flow rate into a confined chamber; (b) flowing a second stream comprising one or more water-soluble polyanionic polymers at a second variable flow rate into the confined chamber; and (c) impinging the first stream and the second stream in the confined chamber until the Reynolds number is from about 1,000 to about 20,000, thereby causing the one or more water-soluble polycationic polymers and the one or more water-soluble polyanionic polymers to undergo a polyelectrolyte complexation process that continuously generates PEC nanoparticles.

2 . The method of claim 1 , wherein the first stream and the second stream are on opposing sides when entering the confined chamber, and wherein the ratio of the flow rate of the second stream to the flow rate of the first stream is from about 1 to about 10.

3 . The method of claim 1 , wherein the first stream and/or the second stream further comprise one or more water-soluble therapeutic agents selected from the group consisting of a small molecule, carbohydrate, sugar, protein, peptide, nucleic acid, antibody or antibody fragment thereof, hormone, hormone receptor, receptor ligand, cytokine, and growth factor.

4 . The method of claim 1 , further comprising flowing a third stream at a third variable flow rate into the confined chamber, wherein each stream is equidistant from the other two streams when entering the confined chamber, and wherein the third stream comprises one or more water-soluble therapeutic agents, one or more miscible organic solvents, and/or one or more cryoprotectants.

5 . The method of claim 4 , wherein the first variable flow rate, the second variable flow rate, and the third variable flow rate if present are greater than about 5 milliliters/minute.

6 . The method of claim 4 , wherein the generated PEC nanoparticles encapsulate the at least one or more water-soluble therapeutic agents.

7 . The method of claim 1 , wherein:
(i) the one or more water-soluble polycationic polymers are selected from the group consisting of chitosan, PAMAM dendrimers, polyethylenimine (PEI), protamine, poly(arginine), poly(lysine), poly(beta-aminoesters), cationic peptides and derivatives thereof; (ii) the one or more water-soluble polyanionic polymers are selected from the group consisting of poly(aspartic acid), poly(glutamic acid), negatively charged block copolymers, heparin sulfate, dextran sulfate, hyaluronic acid, alginate, tripolyphosphate (TPP), oligo(glutamic acid), a cytokine, a protein, a peptide, a growth factor, and a nucleic acid; and (iii) the nucleic acid is selected from the group consisting of an antisense oligonucleotide, cDNA, genomic DNA, guide RNA, plasmid DNA, vector DNA, mRNA, miRNA, piRNA, shRNA, and siRNA.

8 . The method of claim 1 , wherein the Reynolds number ranges from about 2,000 to about 8,000.

9 . The method of claim 1 , wherein the pH value of the first stream and the pH value of the second stream range from about 2.5 to about 8.4.

10 . The method of claim 1 , wherein the generated polyelectrolyte complex (PEC) nanoparticles range in size from about 20 nm to about 500 nm in diameter.

11 . The method of claim 1 , wherein the polydispersity index of the generated polyelectrolyte complex (PEC) nanoparticles ranges from about 0.05 to about 0.1.

12 . The method of claim 1 , wherein:
(i) the one or more water-soluble polyanionic polymers is plasmid DNA and the one or more water-soluble polycationic polymers is selected from the group consisting of linear polyethylenimine (PEI) and its derivatives; or (ii) the first stream comprises chitosan and the second stream comprises tripolyphosphate (TPP) and a protein, wherein the protein is co-encapsulated by the TPP and chitosan in the generated polyelectrolyte complex (PEC) nanoparticles.

13 . The method of claim 1 , wherein the second stream comprises the one or more water-soluble therapeutic agents and the polyelectrolyte complexation process encapsulates the one or more water-soluble therapeutic agents in the generated polyelectrolyte complex (PEC) nanoparticles.

14 . A uniform polyelectrolyte complex (PEC) nanoparticle preparation generated by a flash nanocomplexation (FNC) method of continuously generating uniform polyelectrolyte complex (PEC) nanoparticles, the method comprising:
(a) flowing a first stream comprising one or more water-soluble polycationic polymers at a first variable flow rate into a confined chamber; (b) flowing a second stream comprising one or more water-soluble polyanionic polymers at a second variable flow rate into the confined chamber; and (c) impinging the first stream and the second stream in the confined chamber until the Reynolds number is from about 1,000 to about 20,000, thereby causing the one or more water-soluble polycationic polymers and the one or more water-soluble polyanionic polymers to undergo a polyelectrolyte complexation process that continuously generates PEC nanoparticles.

15 . A device for continuously generating uniform polyelectrolyte complex (PEC) nanoparticles, the device comprising:
(a) a housing comprising: a confined chamber; (ii) a first inlet configured to permit a first stream comprising one or more water-soluble polycationic polymers to flow into the confined chamber; (iii) a second inlet configured to permit a second stream comprising one or more water-soluble polyanionic polymers to flow into the confined chamber; and (iv) optionally a third inlet configured to permit a third stream to flow into the confined chamber; (b) a first conduit providing a first stream path configured to permit the first stream to flow into the confined chamber at a first variable flow rate through the first inlet, wherein the first conduit has a first variable diameter; (c) a second conduit providing a second stream path configured to permit the second stream to flow into the confined chamber at a second variable flow rate through the second inlet, wherein the second conduit has a second variable diameter; and (d) optionally a third conduit providing a third stream path configured to permit the third stream to flow into the confined chamber at a third variable flow rate through the third inlet, wherein the third conduit has a third variable diameter; wherein the first conduit, the second conduit, and the third conduit if present, are situated equidistantly relative to each other on an exterior surface of the housing in a manner that permits impinging of the streams in the confined chamber until the Reynolds number is from about 1,000 to about 20,000, thereby causing the one or more water-soluble polycationic polymers and the one or more water-soluble polyanionic polymers to undergo a polyelectrolyte complexation process that continuously generates PEC nanoparticles.

16 . The device of claim 15 , wherein the confined chamber is a circular cylinder ranging in diameter from about 1.25 mm to about 7.5 mm and ranging in height from about 2.5 mm to about 15 mm.

17 . The device of claim 15 , wherein the first conduit, the second conduit, and the third conduit if present extend from the exterior of the housing, into the inlet, and to the confined chamber.

18 . The device of claim 15 , wherein the first conduit, the second conduit, and the third conduit if present each have a first section with a diamete

Assignees

• Univ Johns Hopkins

Inventors

• Mao Hai-Quan
• Santos Jose Luis
• Ren Yong
• Williford John-Michael

Classifications

• A61K9/5169

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

• A61K47/6455

  Polycationic oligopeptides, polypeptides or polyamino acids, e.g. for complexing nucleic acids · CPC title

• A61K9/5146

  obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, polyamines, polyanhydrides · CPC title

• A61K47/61

  the organic macromolecular compound being a polysaccharide or a derivative thereof · CPC title

• A61K9/5115

  Inorganic compounds · CPC title