Sustained release formulations using non-aqueous emulsions
Non-aqueous emulsion methods for producing polymeric or polymer-coated microparticles are provided. One method produces a sustained release microparticle composition by combining protein powder and a polymer into a hydrocarbon solvent to form a non-aqueous first solution and adding the first solution to a second solution, wherein the second solution comprises a fluorocarbon liquid and a fluorosurfactant to form a non-aqueous emulsion comprising multiple emulsion hydrocarbon droplets in the fluorocarbon liquid. The subsequent microparticle hardening process includes the steps of removing the hydrocarbon solvent from the formed emulsion droplets, which can be achieved through evaporation the hydrocarbon at ambient condition under stirring, or accelerated hardening through vacuum, or through adding hydrofluoroester into the fluorocarbon as a cosolvent. Removing the fluorocarbon liquid and washing with extra fluorocarbon liquid to isolate the sustained release microparticles, wherein the sustained release microparticles comprise one or more cores of protein and a cortex of polymer.
1 . A method of producing encapsulated protein microparticles comprising:
combining spray-dried protein powder and a polymer into a hydrocarbon solvent to form a non-aqueous first solution;
adding the first solution to a second solution, wherein the second solution comprises a fluorocarbon liquid and a fluorosurfactant;
agitating the combined solutions to form a non-aqueous emulsion comprising multiple emulsion hydrocarbon droplets in the fluorocarbon liquid;
removing the hydrocarbon solvent; and
removing the fluorocarbon liquid to isolate the encapsulated protein microparticles, wherein the encapsulated protein microparticles comprise protein encapsulated within a matrix of polymer, wherein the encapsulated protein microparticles are sustained release encapsulated protein microparticles, and wherein the encapsulated protein microparticles comprise a polymer cortex devoid of pores or channels.
2 . The method of claim 1 , wherein the encapsulated protein microparticles comprise a single core-shell structure.
3 . The method of claim 1 , wherein at least one of the encapsulated protein microparticles comprise multiple cores dispersed within the polymer.
4 . The method of claim 1 , wherein the encapsulated protein microparticles comprise microparticles comprising a combination of a single core-structures encapsulated by a polymer and microparticles comprising multi-core structures encapsulated by a polymer.
5 . The method of claim 1 , wherein the fluorocarbon liquid comprises a perfluoro C5-C18 compound.
6 . The method of claim 1 , wherein the hydrocarbon solvent is selected from the group consisting of dichloromethane, chloroform, toluene, ethyl acetate, tetrahydrofuran, or a combination thereof.
7 . The method of claim 1 , wherein the fluorocarbon liquid comprises 1,1,2,2,3,3,4,4,4-nonafluoro-N,N-bis(1,1,2,2,3,3,4,4,4-nonafluorobutyl)butan-1-amine.
8 . The method of claim 1 , wherein the hydrocarbon solvent comprises dichloromethane, ethyl acetate, or a combination thereof.
9 . The method of claim 1 , wherein the fluorocarbon liquid comprises hydrofluoroether.
10 . The method of claim 1 , wherein the fluorosurfactant comprises Perfluoropolyether-b-Polyethylene glycol-b-Perfluoropolyether.
11 . The method of claim 1 , wherein the polymer comprises polyorthoester (POE).
12 . The method of claim 1 , wherein the polymer is selected from the group consisting of polylactic acid and poly(lactic-co-glycolic acid).
13 . The method of claim 1 , wherein the protein is an antibody or antigen binding fragment thereof, a fusion protein, or a recombinant protein.
14 . The method of claim 1 , wherein the protein is a VEGF Trap protein.
15 . The method of claim 14 , wherein the protein is a truncated form of VEGF Trap protein.
16 . The method of claim 1 , wherein the encapsulated protein microparticles have a diameter of 1 to 200 μm.
17 . The method of claim 1 , wherein the protein powder comprises particles comprising a diameter of 0.5 to 20 μm of micronized protein.
18 . The method of claim 1 , wherein protein powder is micronized by spray-drying, electrospray drying, reversible precipitation, spray freezing, microtemplating, or a combination thereof.
19 . The method of claim 1 , wherein the emulsion is formed using homogenization, vortexing, sonication, cavitation, agitation, or a combination thereof.
20 . The method claim 1 , wherein removing the hydrocarbon solvent while stirring the combined solutions.
21 . The method of claim 20 , wherein the hydrocarbon solvent is removed under vacuum to harden the encapsulated protein microparticles.
22 . The method of claim 1 , wherein the hydrocarbon solvent is removed by evaporation.
23 . The method of claim 1 , wherein the fluorocarbon liquid is removed by filtration optionally under vacuum.
24 . The method of claim 22 , wherein a hydrofluoroether is used as a cosolvent to extract the hydrocarbon.
25 . A method for producing encapsulated protein microspheres comprising:
combining
(1) a dispersed phase comprising 1.0 to 30.0% w/w of total solid spray dried-protein suspended in a hydrocarbon solution, wherein the hydrocarbon solution comprises 5.0 to 35% w/v POE, into
(2) a continuous phase to form emulsion droplets of the dispersed phase, wherein the continuous phase comprises a fluorocarbon solution comprising 0.1 to 5.0% w/v fluorosurfactant;
hardening the emulsion droplets by removing the hydrocarbon liquids to form hardened encapsulated protein microspheres, wherein the encapsulated protein microspheres are sustained release encapsulated protein microspheres, wherein the encapsulated protein microspheres comprise a polymer cortex devoid of pores or channels.
26 . The method of claim 25 , wherein the non-aqueous emulsion is stirred, and the hydrocarbon solution is removed by evaporation under ambient atmospheric pressure or under vacuum while stirred.
27 . The method of claim 26 , wherein the hardened encapsulated protein microspheres are harvested by vacuum filtration.
28 . A method for producing encapsulated protein microparticles, comprising:
combining a hydrocarbon solution comprising dissolved polymer and spray-dried protein powder to produce a dispersed phase;
combining the dispersed phase with a continuous phase to produce emulsion droplets of the dispersed phase in the continuous phase, wherein the continuous phase comprises a fluorocarbon liquid and 0.1 to 5.0% w/v of a fluorosurfactant; and
harvesting the encapsulated protein microparticles, wherein the encapsulated protein microparticles are sustained release encapsulated protein microparticles, wherein the encapsulated protein microparticles comprise a polymer cortex devoid of pores or channels.
29 . The method of claim 28 , wherein the spray-dried protein is an antibody, recombinant protein, fusion protein, or a fragment of the spray-dried protein thereof.
30 . The method of claim 28 , wherein the spray-dried protein is a VEGF Trap protein or a truncated VEGF Trap protein.
31 . The method of claim 28 , wherein the hydrocarbon solution is selected from the group consisting of dichloromethane, chloroform, toluene, ethyl acetate, tetrahydrofuran, or a combination thereof.
32 . The method of claim 28 , wherein the fluorocarbon liquid comprises trifluoromethyl)bis(1,1,2,2,3,3,4,4,4-nonafluorobutyl)amine.
33 . The method of claim 28 , wherein the encapsulated protein microparticles are hardened by removing the hydrocarbon solution while stirring by evaporation or under vacuum.
34 . The method of claim 33 , further comprises harvesting the hardened encapsulated protein microparticles.
35 . The method of claim 9 , wherein the hydrofluoroether comprises 4-Ethoxy-1,1,1,2,2,3,3,4,5,6,6,6-didecafluoro-5-(trifluoromethyl)hexane.
36 . A method for producing encapsulated protein microparticles comprising:
combining a first solution comprising a spray-dried protein powder and a polymer in a hydrocarbon solvent with a second solution comprising a fluorocarbon solvent and a fluorosurfactant;
agitating the combined solutions to produce an emulsion;
removing the hydrocarbon solvent under vacuum while stirring the combined solutions to harden the encapsulated protein microparticles;
harvesting the encapsulated protein microparticles;
optionally washing the encapsulated protein microparticles;
and drying the encapsulated protein microparticles, wherein the encapsulated protein microparticles comprises a polymer cortex devoid of pores or channels.
37 . The method of claim 36 , wherein the hydrocarbon solvent is selected from the group consisting of dichloromethane, chloroform, toluene, ethyl acetate, tetrahydrofuran, acetonitrile, ethanol, methanol, propanol, dimethylformamide, dimethyl sulfoxide or a combination thereof.
38 . The method of any one of claim 36 , wherein the fluorocarbon solvent comprises trifluoromethyl)bis(1,1,2,2,3,3,4,4,4-nonafluorobutyl)amine.
39 . The method of claim 36 , wherein the polymer comprises POE, polylactic acid, poly(lactic-co-glycolic acid), or a combination thereof.
40 . The method of claim 36 , wherein the encapsulated protein microparticles comprise a polymer cortex and a hollow core.
41 . The method of claim 36 , wherein a diameter of the encapsulated protein microparticles is tuned to a desired diameter by changing the hydrocarbon solvent, agitation speed, polymer concentration, or a combination thereof.