Method for controlling encapsulation efficiency and burst release of water soluble molecules from nanoparticles and microparticles produced by inverse flash nanoprecipitation

Abstract

A method for controlling the encapsulation efficiency and burst release of water soluble molecules from nanoparticle and microparticle formulations produced by the inverted Flash NanoPrecipitation (iFNP) process and subsequent processing steps is presented. The processing steps and materials used can be adjusted to tune the encapsulation efficiency and burst release of the encapsulated water-soluble material. The encapsulation efficiency of the soluble agent in the particles and the burst release of the soluble agent from the particles can be controlled by: (1) the copolymers used in the assembly or coating process, (2) the degree of crosslinking of the nanoparticle core, (3) the incorporation of small molecule or polymeric additives, and/or (4) the processing and release conditions employed.

1 . A method of forming a polymer inverse nanoparticle that encapsulates a water soluble active to maximize or optimize encapsulation efficiency and/or to minimize or optimize burst fraction, comprising:
 dissolving the water soluble active at a concentration and a block copolymer at a concentration in an amount of a process solvent to form a process solution; and   continuously mixing the process solution with an amount of a nonprocess solvent at a process temperature to form a first nanoparticle solution comprising polymer inverse nanoparticles having a core and a shell and a first nanoparticle solvent;   wherein the block copolymer comprises a hydrophilic block and a hydrophobic block having a glass transition temperature (Tg),   wherein the hydrophilic block is soluble in the process solvent and is insoluble in the nonprocess solvent,   wherein the hydrophobic block is insoluble in the process solvent and is soluble in the nonprocess solvent,   wherein the process solution is more polar than the nonprocess solvent,   wherein the water soluble active and the hydrophilic block are in the core and the hydrophobic block is in the shell, and   wherein the encapsulation efficiency is maximized or optimized by
 (a) selecting the process solvent, so that the hydrophilic block is close to a solubility limit in the process solution for the concentration of the block copolymer, and/or 
 (b) crosslinking the hydrophilic block in the core, and/or 
 (c) selecting the hydrophilic block to have bonding interactions with the water soluble active in the core, and/or 
 (d) selecting the hydrophobic block to have a molecular weight of at least 10 kDa, 20 kDa, 30 kDa, 40 kDa, 50 kDa, 70 kDa, or 100 kDa, and/or 
 (e) selecting the process temperature and/or the hydrophobic block, so that the process temperature is less than the hydrophobic block glass transition temperature (Tg), and/or 
 (f) selecting the process solvent to have high osmolarity (e.g., by dissolving a salt in the process solvent), and/or 
 (g) adding a supplemental hydrophobic compound to the process solvent and/or to the nonprocess solvent to increase the bulk of hydrophobic material in the shell, 
   and/or   wherein the burst fraction is minimized or optimized by
 (aa) crosslinking the hydrophilic block in the core, and/or 
 (bb) increasing the hydrophobic block glass transition temperature (T g ), and/or 
 (cc) adding a supplemental hydrophobic compound to the process solvent and/or to the nonprocess solvent to increase the bulk of hydrophobic material in the shell. 
   
     
     
         2 . The method of  claim 1 ,
 wherein the encapsulation efficiency is optimized by crosslinking the hydrophilic block in the core,   wherein the burst fraction is minimized by crosslinking the hydrophilic block in the core, and   wherein the crosslinking agent is selected from the group consisting of a metal, calcium, a chelating agent, tetraethylene pentamine (TEPA), and combinations.   
     
     
         3 . The method of  claim 1 ,
 wherein the encapsulation efficiency is maximized by adding a supplemental hydrophobic compound to the nonprocess solvent to increase the bulk of hydrophobic material in the shell,   wherein the burst fraction is minimized by adding a supplemental hydrophobic compound to the nonprocess solvent to increase the bulk of hydrophobic material in the shell, and   wherein the supplemental hydrophobic compound is selected from the group consisting of a hydrophobic polymer, polylactic acid, vitamin E, and combinations.   
     
     
         4 . The method of  claim 1 , wherein the burst fraction is minimized by selecting the process temperature and/or the hydrophobic block, so that the process temperature is less than the hydrophobic block glass transition temperature (Tg). 
     
     
         5 . The method of  claim 1 , further comprising
 annealing the polymer inverse nanoparticle,   wherein the annealing maximizes the encapsulation efficiency and   wherein the annealing optimizes the encapsulation efficiency.   
     
     
         6 . The method of  claim 1 , further comprising adding lecithin to the nonprocess solvent. 
     
     
         7 . The method of  claim 1 , wherein the water soluble active is selected from the group consisting of a linear polypeptide, a cyclic polypeptide, ovalbumin, lysozyme, PEP1, and vancomycin. 
     
     
         8 . The method of  claim 1 ,
 wherein the hydrophilic block is selected from the group consisting of poly(aspartic acid) and poly(glutamic acid) and   wherein the hydrophobic block is selected from the group consisting of poly(lactic acid), poly(lactic-co-glycolic acid), and poly(caprolactone).   
     
     
         9 . The method of  claim 1 ,
 wherein the hydrophilic block is of a molecular weight in the range of from 0.2 kDa to 100 kDa, from 0.5 kDa to 50 kDa, from 1 kDa to 20 kDa, from 2 kDa to 10 kDa, or of about 5 kDa, and   wherein the hydrophobic block is of a molecular weight in the range of from 0.5 kDa to 400 kDa, 1 kDa to 200 kDa, from 2 kDa to 100 kDa, from 5 kDa to 100 kDa, from 10 kDa to 40 kDa, of about 10 kDa, of about 20 kDa, or of about 40 kDa.   
     
     
         10 . The method of  claim 1 , wherein the supplemental hydrophobic compound is selected from the group consisting of poly(lactic acid), poly(lactic-co-glycolic acid), and poly(caprolactone). 
     
     
         11 . The method of  claim 1 , wherein the process solvent is miscible with the nonprocess solvent. 
     
     
         12 . The method of  claim 1 , further comprising
 adding a second block copolymer to the first nanoparticle solution to form a second stage process solution; and   continuously mixing the second stage process solution with a finishing solvent to form a second nanoparticle solution comprising the polymer inverse nanoparticles coated with the second block copolymer,   wherein the second block copolymer comprises a second hydrophilic block and a second hydrophobic block.   
     
     
         13 . The method of  claim 12 ,
 wherein the second hydrophilic block is selected from the group consisting of poly(ethylene glycol) and poly(propylene oxide) and   wherein the second hydrophobic block is selected from the group consisting of poly(lactic acid), poly(lactic-co-glycolic acid), and poly(caprolactone).   
     
     
         14 . The method of  claim 12 ,
 wherein the second hydrophilic block is of a molecular weight in the range of from 0.2 kDa to 100 kDa, from 0.5 kDa to 50 kDa, from 1 kDa to 20 kDa, from 2 kDa to 10 kDa, or of about 5 kDa and   wherein the second hydrophobic block is of a molecular weight in the range of from 0.2 kDa to 100 kDa, from 0.5 kDa to 50 kDa, from 1 kDa to 20 kDa, from 2 kDa to 10 kDa, or of about 5 kDa.   
     
     
         15 . The method of  claim 12 , wherein the second stage process solution is miscible with the finishing solvent. 
     
     
         16 . The method of  claim 1 , further comprising
 concentrating the polymer inverse nanoparticles to form microparticles,   wherein each microparticle comprises a plurality of nanoparticles.   
     
     
         17 . The method of  claim 1 ,
 wherein the water soluble active is anionic and the hydrophilic block is selected to be cationic, so that water soluble active and the hydrophilic block ionically bond or   wherein the water soluble active is cationic and the hydrophilic block is selected to be anionic, so that water soluble active and the hydrophilic block ionically bond.   
     
     
         18 . The method of  claim 1 , further comprising adding a tackifier to the process solvent and/or to the nonprocess solvent to increase the hydrophobic block glass transition temperature (Tg). 
     
     
         19 . The method of  claim 1 ,
 wherein the process solvent and the finishing solvent are each independently selected from the group consisting of dimethylsulfoxide (DMSO), propanol, ethanol, methanol, water, and combinations and   wherein the nonprocess solvent is selected from the group consisting of dichloromethane, chloroform, acetone, tetrahydrofuran (THF), methanol, and combinations.   
     
     
         20 . The method of  claim 1 , wherein the continuous mixing is through a flash nanoprecipitation process. 
     
     
         21 . A polymer inverse nanoparticle that encapsulates a water soluble active, comprising
 a triblock copolymer comprising two hydrophilic end blocks and a hydrophobic center block;   a core; and   a shell,   wherein the hydrophobic center block is between each of the two hydrophilic end blocks,   wherein the water soluble active and the hydrophilic end blocks are within the core,   wherein the hydrophobic center block is within the shell, and   wherein the hydrophilic end blocks are crosslinked within the core with a crosslinking agent.   
     
     
         22 . The polymer inverse nanoparticle of  claim 21 , wherein the hydrophobic end blocks are formed from the same monomer. 
     
     
         23 . The polymer inverse nanoparticle of  claim 21 , wherein the water soluble active comprises a linear polypeptide, a cyclic polypeptide, ovalbumin, lysozyme, PEP1, or vancomycin. 
     
     
         24 . The polymer inverse nanoparticle of  claim 21 ,
 wherein each hydrophilic end block is independently selected from the group consisting of poly(aspartic acid) and poly(glutamic acid) and   wherein the hydrophobic center block is selected from the group consisting of poly(lactic acid), poly(lactic-co-glycolic acid), and poly(caprolactone).   
     
     
         25 . The polymer inverse nanoparticle of  claim 21 , wherein the triblock copolymer is selected from the group consisting of poly(aspartic acid)-b-poly(lactic acid)-b-poly(aspartic acid), poly(glutamic acid)-b-poly(lactic acid)-b-poly(glutamic acid), poly(aspartic acid)-b-poly(lactic-co-glycolic acid)-b-poly(aspartic acid), and poly(glutamic acid)-b-poly(lactic-co-glycolic acid)-b-poly(glutamic acid). 
     
     
         26 . The polymer inverse nanoparticle of  claim 21 , further comprising
 a diblock copolymer comprising a hydrophilic block and a hydrophobic block; and   a coating comprising an interior layer and an exterior layer,   wherein the hydrophobic block is within the interior layer,   wherein the hydrophilic block is within the exterior layer, and   wherein the interior layer is adjacent to the shell.   
     
     
         27 . The polymer inverse nanoparticle of  claim 26 ,
 wherein the hydrophilic block is polyethylene glycol (PEG) and   wherein the hydrophobic block is polylactic acid (PLA).