Compositions and methods for reducing agglomeration of microparticles in coating processes

Abstract

Embodiments of the present disclosure provide novel compositions and methods for maintaining particle or microparticle separation and / or reducing or preventing particle or microparticle agglomeration in a coating process of a chemical vapor deposition system, chemical layer deposition system, atomic layer deposition (ALD) system, and the like. In certain embodiments, compositions and methods include mixing one or more types of milling agents with thermostable therapeutic agent-, antigen-, immunogen-, compound-containing particles or adding one or more types of milling agents into a coating chamber of a reactor particle-coating system for coating therapeutic agent-, antigen-, immunogen-, compound-containing particles with one or more metal and / or oxide coating layer. In other embodiments, a milling screen is introduced to a coating system disclosed herein to separate milling agents from therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles during coating processes.

Description

COMPOSITIONS AND METHODS FOR REDUCING AGGLOMERATION OF MICROPARTICLES IN COATING PROCESSES PRIORITY

[0001] This International Application claims priority to U.S. Provisional Application No.63 / 752,355, entitled, “Compositions and Methods for Reducing Agglomeration of Microparticles in Coating Processes,” filed January 31, 2025. This provisional application is incorporated by reference in its entirety for all purposes.FIELD

[0002] Embodiments of the present disclosure provide novel methods for maintaining particle or microparticle separation and / or reducing or preventing particle or microparticle agglomeration during a process to deposit coating layers on the particle or microparticle surfaces using a chemical vapor deposition system, a chemical layer deposition system, an atomic layer deposition (ALD) system, and the like. In certain embodiments, compositions and methods include mixing one or more types of milling agents with thermostable therapeutic agentcontaining particles or adding one or more types of milling agents into a coating chamber of a deposition system for coating for improved and consistent coating of the particles or microparticles.BACKGROUND

[0003] Atomic Layer Deposition (ALD) is a gas or vapor-phase process where thin conformal shells of chemical compounds are grown atomic layer by atomic layer on the surfaces of particles including but not limited to nanoparticles and microparticles housing targeted agents (e g., therapeutic agents or other agent or compound) of stabilized formulations using repeated multi-step reaction processes to create thin conformal shells coating the surface of particles or microparticles. Generally, chemical precursors or agents (e.g., biological, chemical, microbial, or pharmaceutical agents) are generated into thermostable dry particles so that all surface sites of these particles are then evenly exposed throughout a coating process and uniform coatings are obtained. As these coated microparticles or particles are formed, particles can stick to other particles forming difficult to disperse agglomerates or clusters in the bulk phase end product that can increase in size with increasing processing time and deposition layering. As the coating process proceeds, agglomerates can solidify by binding adjoining particles with increasingcoating thickness. This agglomeration results in the points of contact between particles coating a targeted agent or stabilized formulation being left unexposed to the chemical precursors intended to form uniform coatings on the particles or microparticles. Therefore, when the agglomerates are redispersed (e.g., particles are detached from one another), contact points can provide weak and uneven sections of coating or no coating at all and, in certain cases, the contact points can form holes in the coating allowing the target agents or stabilized formulations to leak out prematurely which results in loss of the desired fully coated particles and loss of product.Therefore, there is a need for improved systems for reducing or preventing agglomeration or particle adherence during these processes.SUMMARY

[0004] Embodiments of the present disclosure provide compositions and methods for coating pharmaceutical agent-containing particles or microparticles in a chemical vapor deposition system with minimal agglomeration. In some embodiments, methods disclosed herein can include introducing a plurality of milling agents into a coating chamber of the chemical vapor deposition system, introducing a plurality of stabilized pharmaceutical agent-containing particles or microparticles to the coating chamber of the chemical vapor deposition system, and coating the plurality of stabilized pharmaceutical agent-containing particles or microparticles with at least one of a metal agent (e.g. metal oxide or metal alkoxide), and / or other chemical layer, forming the minimally agglomerated coated plurality of pharmaceutical agent-containing particles or microparticles. In some embodiments, coating of the plurality of pharmaceutical agent-containing particles or microparticles can be conducted under vacuum or elevated pressures using chemical vapor deposition methods. In some embodiments, coating of the plurality of pharmaceutical agent-containing particles or microparticles can be performed using atomic layer deposition (ALD).

[0005] In certain embodiments and further to paragraph

[0004] above, methods can further include mechanically agitating the plurality of milling agents and the plurality of pharmaceutical agent-containing particles or microparticles within the coating chamber of the chemical vapor deposition system during coating of the plurality of pharmaceutical agent-containing particles or microparticles. In accordance with these embodiments, a plurality of milling agents and a plurality of pharmaceutical agent-containing particles or microparticles can be agitated by at least one of: a flowing gas stream, mechanical vibration of the coating chamber, rotary motion ofthe coating chamber, magnetic field agitation in case of ferromagnetic particles, stirrers or impellers in the coating chamber bed, inducing sufficient mechanical movement to continuously break apart agglomerates of the plurality of pharmaceutical agent-containing particles or microparticles to be coated. In some embodiments, agitation can include mechanical vibration of the coating chamber using a sonication device associated with the coating chamber.

[0006] In certain embodiments and further to paragraphs

[0004] -

[0005] above, introducing the plurality of milling agents and introducing the plurality of pharmaceutical agent-containing particles or microparticles, to the coating chamber of the chemical vapor deposition system can include premixing and / or homogenizing the plurality of the milling agents and the plurality of pharmaceutical agent-containing particles or microparticles prior to introducing them to the coating chamber of the chemical vapor deposition system. In certain embodiments, introducing the plurality of milling agents and introducing the plurality of pharmaceutical agent-containing particles or microparticles, to the coating chamber of the chemical vapor deposition system can also include simultaneously introducing the plurality of the milling agents and the plurality of pharmaceutical agent-containing particles or microparticles. In other embodiments, introducing the plurality of milling agents and introducing the plurality of pharmaceutical agent-containing particles or microparticles to the coating chamber of the chemical vapor deposition system can also include separately introducing the plurality of the milling agents and the plurality of pharmaceutical agent-containing particles or microparticles to the coating chamber.

[0007] In other embodiments and further to paragraphs

[0004] -

[0006] above, minimally agglomerated coated pharmaceutical agent-containing particles or microparticles and plurality of milling agents can be removed from the coating chamber after up to 10 coating cycles, up to 20 coating cycles, or up to 30 coating cycles or more. In some embodiments, the minimally agglomerated coated pharmaceutical agent-containing particles or microparticles can be reintroduced to the coating chamber for additional coatings, as desired and for additional number of coatings desired (e.g., up to 250, up to 500 or more). In certain embodiments, the plurality of milling agents and the minimally agglomerated coated pharmaceutical agent-containing particles or microparticles disclosed herein can be removed from the coating chamber of the chemical vapor deposition system after up to 10 coating cycles, up to 20 coating cycles, or up to 30 coating cycles or more coating layers are applied to the pharmaceutical agent-containing particles or microparticles before reintroducing the coated pharmaceutical agent-containingparticles or microparticles to the coating chamber and depositing at least one final coating layer onto the plurality of pharmaceutical agent-containing particles or microparticles in the absence of the plurality of milling agents. In accordance with these embodiments, the milling agents can be removed from the chamber and not reintroduced in part because the particles are fully coated with initial coating layers and have a significantly reduced chance of agglomerating and removal of the milling agents at an intermediate period of coating eliminates the milling agents from a final coated particle product.

[0008] In certain embodiments and further to paragraphs

[0004] -

[0007] above, when removed from a coating chamber of a coating system of use herein, the plurality of milling agents can be separated from partially or fully coated pharmaceutical agent-containing particles or microparticles. In certain embodiments, the plurality of milling agents can be separated from the partially or fully coated pharmaceutical agent-containing particles or microparticles using a separation device or other manner. In some embodiments, separation can be based on physical properties including but not limited to at least one of size, density, magnetic, ferromagnetic, other properties or a combination thereof. In other embodiments, either within the coating chamber or outside of the coating chamber, the plurality of milling agents and the plurality of coated pharmaceutical agent-containing particles or microparticles can be separated by sieving, fluid dynamic methods, or using magnetic forces from the plurality of milling agents or other suitable methods. In certain embodiments, no milling agents remain in contact with the partially or fully coated pharmaceutical agent-containing particles or microparticles once a separation technique is applied (e.g., magnetic separation).

[0009] In certain embodiments and further to paragraphs

[0004] -

[0008] above, a separated partially or fully coated minimally agglomerated plurality of pharmaceutical agent-containing particles or microparticles can further be stored for later use or form part of an essentially dry or rehydrated pharmaceutical composition. In some embodiments, the pharmaceutical composition can when rehydrated can further include at least one pharmaceutically acceptable excipient or carrier. In some embodiments, separated minimally agglomerated partially coated plurality of pharmaceutical agent-containing particles or microparticles can be re-introduced into the coating chamber for additional coatings. In some embodiments, pharmaceutical agent-containing particles or microparticles can be coated with at least 10 to about 2,000 coating cycles or more, as desired.

[0010] In certain embodiments and further to paragraphs

[0004] -

[0009] above, the coating chamber can be any one of a fluidized bed reactor, a gas-phase powder reactor, or a rotary reactor. In some embodiments, the coating chamber is a fluidized bed reactor of an ALD system.

[0011] In some embodiments and further to paragraphs

[0004] -

[0010] above, milling agents of the plurality of milling agents contemplated herein can include one or more of inorganic and organic milling agents. In certain embodiments, the one or more of inorganic and organic milling agents can include, but are not limited to, one or more of zirconium dioxide (ZrCh), zirconium dioxide / silicon dioxide (ZrCh / SiCh), iron (Fe), aluminum oxide (AI2O3), glass, organic polymers (e.g., polystyrene, Teflon, polyethylene, Nylon, starch, cellulose or the like), or other similar solid material or combinations thereof. In some embodiments, milling agents of use in composition and methods and systems disclosed herein include at least one metal agent (e.g. metal oxide or metal alkoxide) or non-metal agent. In accordance with these embodiments, the at least one metal milling agent can include, but is not limited to, at least one metallo-organic material, metal oxide, metal alkoxide, or a combination thereof. In some embodiments, the non-metal agents can include non-metal oxide. In some embodiments, the at least one metal oxide can include at least one of aluminum oxide, aluminum alkoxide, titanium dioxide (TiCh), zinc oxide (ZnO), and any combinations thereof. In other embodiments, the at least one milling agent can include at least zirconium dioxide / silicon dioxide (ZrCh / SiCh). In some embodiments, the at least one non-metal oxide can include at least silicon dioxide. In certain embodiments, milling agents of use in compositions and methods and systems disclosed herein can include at least one large, spray-dried milling media particle with a mass greater than the particles to be coated and large enough to be separated from the particles to be coated, created using the same formulation as the particles to be coated.

[0012] In certain embodiments and further to paragraphs

[0004] -

[0011] above, the present disclosure provides methods for coating pharmaceutical agent-containing particles or microparticles in a chemical vapor deposition system. In some embodiments, methods can include, but are not limited to, introducing a plurality of milling agents onto a milling screen (e.g., mesh), where the milling screen is above a fluidized bed in a coating chamber of the chemical vapor deposition system, introducing a plurality of pharmaceutical agent-containing particles or microparticles to the fluidized bed in the coating chamber of the chemical vapor deposition system, and coating the plurality of pharmaceutical agent-containing particles ormicroparticles with at least one of metal agent (e.g. metal oxide or metal alkoxide) forming a minimally agglomerated plurality of coated pharmaceutical agent-containing particles or microparticles where the milling agents are separated from bulk of the pharmaceutical agentcontaining particles or microparticles by the milling screen. In some embodiments, a fraction of a particle or microparticle-containing powder (e.g., bulk powder or dehydrated, spray-dried particles) including some agglomerated particles or microparticles, can be aerosolized by process gases of a coating system disclosed herein and can intermittently contact agitated or mobile milling agents during a coating process to induce deagglomeration before recombining these particles with a bulk powder phase. In other embodiments, a milling screen can be separated from a base of the coating chamber containing a plurality of pharmaceutical agent-containing particles or microparticles on a gas outlet side (versus a gas inlet side) of the plurality of pharmaceutical agent-containing particles or microparticles to be coated and milling agents introduced on an outlet side of the milling screen, away and separated from a plurality of pharmaceutical agent-containing particles or microparticles. In certain embodiments, methods can include mechanically agitating the coating chamber during coating of the plurality of pharmaceutical agent-containing particles or microparticles with the at least one of metal agent (e.g. metal oxide or metal alkoxide) or other coating agent in the presence of the milling agents. In some embodiments, the plurality of milling agents can be removed from the coating chamber after up to 10, up to 20, or up to 30 or more coating cycles of the particles and the milling screen can remain or be removed as desired. In some embodiments, the milling agents can be separated from these coated or intermediary coated particles (e.g., where particles can be reintroduced to a reaction chamber for more coatings without the milling agents being present) using methods disclosed herein and not reintroduced to the reaction chamber with particles for more coating.

[0013] In certain embodiments and further to paragraphs

[0004] -

[0012] above, a milling screen can include a mesh or separation feature to separate a plurality of milling agents from the pharmaceutical agent-containing particles or microparticles while permitting intermittent contact of the plurality of milling agents with the pharmaceutical agent-containing particles or microparticles during a coating process. In some embodiments, the mesh or separation feature can include at least one of a wire mesh, or plastic mesh, or any other milling screen. In some embodiments, the milling screen can further contain pores, a mesh, or a consistent pattern open to particles of size smaller than an opening to freely pass through. In certain embodiments, amesh pore size can range from about 1.0 micron to about 200.0 microns or from about 50.0 to about 100.0 microns in diameter or square side length or opening, or other pattern. In some embodiments, a mesh pore size can be about 75.0 microns. In certain embodiments, size of the openings in the milling screen can be adjusted to an appropriate size to reduce or prevent milling agent transfer from one location to another in a coating chamber.

[0014] In certain embodiments and further to paragraphs

[0004] -

[0013] above, the milling agents of the plurality of milling agents can include solid agents. In some embodiments, the milling agents of the plurality of milling agents can include one or more of inorganic or organic milling agents. In some embodiments, the milling agents can include one or more of zirconium dioxide (ZrCh), zirconium dioxide / silicon dioxide (ZrCh / SiCh), iron (Fe), or aluminum oxide (AI2O3), glass, or other similarly solid material, or combination materials thereof. In some embodiments, the milling agents can include a three-dimensional shape; optionally, including a sphere, a cube, a cone, a pyramid, or other shape of uniform milling agents or a mixture of shapes or mixture of irregular shapes. In certain embodiments, a mass ratio of the plurality of milling agents to the plurality of pharmaceutical agent-containing particles or microparticles can range from about 1:10 up to about 50:1, optionally from about 1:1 to about 3:1. In certain embodiments, the milling agents can be about 2 to about 10 times larger than an average size of the plurality of pharmaceutical agent-containing particles or microparticles. In some embodiments, the plurality of milling agents can include one or more inorganic materials or one or more organic material.

[0015] In certain embodiments and further to paragraphs

[0004] -

[0014] above, milling agents of the plurality of milling agents can have a density, mass, and size higher or greater than both the density, mass, and size of pharmaceutical agent-containing particles or microparticles of the plurality of pharmaceutical agent-containing particles or microparticles and minimally agglomerated coated pharmaceutical agent-containing particles or microparticles of the minimally agglomerated plurality of coated pharmaceutical agent-containing particles or microparticles. In some embodiments, a larger density, mass, and / or size of the milling agents reduces or prevents the milling agents from being elutriated from the milling screen by drag of the process gas whereas the process gas flow can be sufficient to elutriate and carry a fraction of the pharmaceutical agent-containing particles and agglomerates through a bed of milling agents place on a milling screen, allowing intermittent contact with the pharmaceutical agent-containing particles or microparticles before resettling into the bulk powder bed to reduce or preventagglomeration during coating processes of the plurality of pharmaceutical agent-containing particles or microparticles.

[0016] In certain embodiments and further to paragraphs

[0003] -

[0015] above, a diameter of milling agents of the plurality of milling agents can be larger or greater than a pore size of the mesh or separation feature disclosed herein. In certain embodiments, the diameter can be at least about 1.1 to about 3.0 times larger or greater than the pore size of the mesh or separation feature.

[0017] In certain embodiments and further to paragraphs

[0004] -

[0016] above, the plurality of milling agents can be removed from the chemical vapor deposition system after about 10, about 20, about 30 or more coating cycles. In accordance with these embodiments, after exposure to the plurality of milling agents, the plurality of pharmaceutical agent-containing particles or microparticles become a population of e a minimally agglomerated partially or fully coated plurality of pharmaceutical agent-containing particles or microparticles. In some embodiments, one or more coating layers of particle contemplated herein can include, but is not limited to, one or more coating layer of aluminum oxide (AI2O3), an aluminum alkoxide, silicon dioxide (SiCh), Zinc dioxide (ZnCh), titanium dioxide (TiCh), and silicon nitride (S13N1) or combinations thereof or alternating layers thereof. In accordance with these embodiments, one, two, three, four, five, up to ten, up to 20, up to 50, up to 100, up to 150, up to 200, up to 500 or more coating layers can encase one or more therapeutic agent- or compound-containing particles disclosed herein. In certain embodiments, the plurality of pharmaceutical agent-containing particles or microparticles can be further coated with at least one of the metal agent (e g. metal oxide) after removal of the plurality of milling agents.

[0018] In certain embodiments and further to paragraphs

[0004] -

[0017] above, a fully coated plurality of pharmaceutical agent-containing particles or microparticles can be removed from the coating chamber and further be stored for later use or form part of a composition or pharmaceutical composition. In accordance with these embodiments, the plurality of milling agents has been removed from a composition including a plurality of pharmaceutical agentcontaining particles or microparticles by methods disclosed herein.

[0019] In certain embodiments and further to paragraphs

[0004] -

[0018] above, the present disclosure includes an ALD system for coating a plurality of pharmaceutical agent-containing particles and microparticles in the presence of a plurality of milling agents contemplated herein. In accordance with these embodiments, the ALD system has a coating chamber configured tohouse the plurality of pharmaceutical agent-containing particles or microparticles and in certain embodiments, a plurality of milling agents therein and configured to permit a flow of process gas therethrough to coat the plurality of pharmaceutical agent-containing particles and microparticles, a gas inlet connected to the coating chamber on one end, a gas outlet connected to the coating chamber on an opposite end of the gas inlet, and one or more agitator(s) associated with or coupled to the coating chamber and configured to deliver mechanical energy to the coating chamber. In some embodiments, the one or more agitator(s) can be a sonicator configured to deliver mechanical energy through a wave propagating structure coupled to the sonicator and the coating chamber. In other embodiments, the agitators can be mechanical impactors and / or vibrating motors coupled to the coating chamber. In some embodiments, the coating chamber is positioned between the gas inlet and the gas outlet of the coating system (e.g., ALD). In some embodiments, the coating chamber can be configured with a milling screen onto which a plurality of milling agents is placed and separated from a plurality of pharmaceutical agent-containing particles or microparticles. In some embodiments, the milling screen can include a mesh or separation feature. In some embodiments, the mesh can be positioned above the fluidized bed within the coating chamber. The distance of placement of the mesh between the fluidized bed and the milling screen can vary. In some embodiments, a plurality of therapeutic agent-containing particles or microparticles forming a powder bed (e.g., spray-dried or evaporatively dried particles) on a fluidized bed in a coating system disclosed herein can expand during fluidization of the plurality of pharmaceutical agent-containing particles or microparticles because of process gas and a milling screen or mesh disclosed herein can be positioned sufficiently high above the plurality of therapeutic agent-containing particles and microparticles to be coated to permit freedom of movement of the plurality of therapeutic agent-containing particles and microparticles beneath the milling screen or mesh, even after accounting for expansion during fluidization of the particles and microparticles. In accordance with these embodiments, this can be due to the process gas and as such allows for periodic contact of temporarily elutriated particles with the plurality of milling agent through the mesh or milling screen. In some embodiments, the plurality of milling agents can be placed on an opposite side of the mesh from the plurality of pharmaceutical agent-containing particles and microparticles positioned within the coating chamber. In some embodiments, the size of the pores of the mesh can be sufficiently large to allow elutriating and recirculating of the plurality of pharmaceuticalagent-containing particles and microparticles within the coating chamber during a coating process without loss of desired coated product.

[0020] In certain embodiments and further to paragraphs

[0004] -

[0019] above, the present disclosure provides kits including, but not limited to, a plurality of milling agents, a plurality of therapeutic agent-containing particles and microparticles, and instructions for using a chemical vapor deposition system; optionally a mesh or milling screen. In some embodiments, kits can include 2 or more types of milling agents or a mixture of milling agents or milling agents of mixed materials and a plurality of therapeutic agent-containing particles and microparticles to be coated.

[0021] In certain embodiments and further to paragraphs

[0004] -

[0020] above, devices for use in coating a plurality of pharmaceutical agent-containing particles and microparticles in the presence of a plurality of milling agents are disclosed. In accordance with these embodiments, a device can include a cylindrical chamber having chamber walls forming a coating chamber operable to house a plurality of pharmaceutical agent-containing particles or microparticles, an inlet at a first end of the coating chamber, an outlet at a second end (e.g., opposite end) of the coating chamber, and a milling screen disposed between the inlet and the outlet within the coating chamber to maintain some separation of a plurality of milling agents or beads from the plurality of pharmaceutical agent-containing particles or microparticles. In some embodiments, the coating chamber can be configured to permit a flow of process gas to pass there through from the inlet to the outlet to coat the plurality of pharmaceutical agent-containing particles or microparticles to undergo a coating process in a fluidized bed environment. In accordance with these embodiments, a milling screen as described herein can be positioned within the coating chamber but above a bulk phase or bulk powder (e.g., plurality of dried particles and microparticles) of a fluidized particle bed to maintain separation of milling agents from the bulk phase or bulk powder being coated of the fluidized particle bed.

[0022] In certain embodiments and further to paragraphs

[0004] -

[0021] above, the present disclosure is directed to a mixture of a plurality of pharmaceutical agent-containing particles or microparticles and a plurality of milling agents. In some embodiments, the mixture can be essentially dry such as a powder or particle material. In certain embodiments, the plurality of pharmaceutical agent-containing particles or microparticles are partially or fully coated with a metal agent (e.g. metal oxide or metal alkoxide) contemplated herein.

[0023] In certain embodiments and further to paragraphs

[0004] -

[0022] above, the plurality of pharmaceutical agent-containing particles or microparticles can be formed by encasing the pharmaceutical agent in a glassy matrix using at least one glass-forming agent and a particle forming process such as spray-drying or other evaporative drying technique known in the art. In some embodiments, the at least one glass-forming agent can include one or more of trehalose, sucrose, ficoll, dextran, maltotriose, lactose, hydroxyethyl starch, glycine, glycine and mannitol, cyclodextrin, povidone, or a combination thereof. In some embodiments, the plurality of pharmaceutical agent-containing particles or microparticles may include spray-dried, lyophilized, vacuum-dried, or spray-freeze-dried plurality of glassy pharmaceutical agent-containing particles or microparticles. In some embodiments, the plurality of pharmaceutical agent-containing particles or microparticles can be minimally agglomerated particles or microparticles, optionally less than 1.0% of the pharmaceutical agent-containing particles or microparticles can be agglomerated compared to a mixture not including the plurality of milling agents during coating processes. In some embodiments, the plurality of pharmaceutical agent-containing particles or microparticles can include at least one coating layer applied by chemical vapor deposition. In some embodiments, the chemical vapor deposition can include ALD.

[0024] In certain embodiments and further to paragraphs

[0004] -

[0023] above, the pharmaceutical agent of the plurality of pharmaceutical agent-containing particles or microparticles can include at least one of immunogenic agents, small molecule therapeutics, anticancer agents, anti-inflammatory agents, anti-immune agents, polynucleotide or complex thereof, polypeptides, antibodies or fragments thereof, other therapeutics, other compounds and / or other biologies and combinations thereof. In some embodiments, the therapeutic agents can include, but are not limited to a viral antigen, bacterial antigen, toxin, fungal agent, recombinant peptide, recombinant protein, peptide derived from a target protein or pathogen, synthetic peptide or protein, a polynucleotide or fragment thereof, or combination thereof. In some embodiments, the therapeutic agent can include at least one of a virus, a virus-like particle, a live virus, a live attenuated virus, an inactivated virus, a bacterial antigen, a bacteriophage, a phage, a prion, or a combination thereof. In some embodiments, the pharmaceutical agent of the plurality of pharmaceutical agent-containing particles or microparticles can include at least one polynucleotide and the at least one polynucleotide can include, but is not limited to, mRNA,DNA, siRNA, gRNA, snRNA, other RNA, a chimera or chimeric polynucleotides or the like, or fragment thereof or combination thereof.

[0025] In certain embodiments and further to paragraphs

[0004] -

[0024] above, the present disclosure is directed to a composition created using processes disclosed herein. In accordance with these embodiments, the composition can include a mixture of minimally agglomerated, coated plurality of pharmaceutical agent-containing particles or microparticles. In some embodiments, the plurality of pharmaceutical agent-containing particles or microparticles can form a core for coating and further be in contact with a plurality of milling agents for initial (e.g., up to 30 coating layers or more) or all coating stages a chemical deposition process disclosed herein.BRIEF DESCRIPTION OF THE DRAWINGS

[0026] The accompanying drawings are incorporated into and form a non-limiting part of the specification to illustrate several examples of the present disclosure.

[0027] FIGS. 1 A-1C represent ALD systems in accordance with certain embodiments disclosed herein. FIG. 1 A represents a cross-sectional view illustrating an ALD system in accordance with certain embodiments disclosed herein. FIGS. 1B-1C are photographs of a disassembled reactor vessel (FIG. IB) and assembled reactor vessel (FIG. 1C) illustrating positioning for a mesh or a mesh in accordance with certain embodiments disclosed herein.

[0028] FIGS. 2A-2C represent cross-sectional views illustrating the placement of a mesh in the coating chamber of a mechanically agitated reactor in accordance with certain embodiments disclosed herein. In FIG. 2A, the mesh is in the middle of the coating chamber in accordance with certain embodiments disclosed herein. In FIG. 2B, the mesh is closer to the outlet of the coating chamber in accordance with certain embodiments disclosed herein. In FIG. 2C, the mesh is closer to the inlet of the coating chamber in accordance with certain embodiments disclosed herein.

[0029] FIGS. 3A-3B represent cross-sectional views illustrating a coating chamber of devices and uses herein having three components in accordance with certain embodiments disclosed herein. These three components include a top component, a milling screen, and a bottom component, the milling screen associated with either the bottom of the top component or the top of the bottom component in this configuration. The top and bottom components can furtherinclude one or more of a frit, and / or filter as depicted according to some embodiments disclosed herein.

[0030] FIG. 4 is a representative graph illustrating normalized o-phtalaldehyde (OP A) dissolution of spray -dried and ALD coated spray-dried powder (100 AI2O3 cycles) in buffer as function of time according to one embodiment of the present disclosure. Squares (closed) represent coated without milling agents. Circles (open) represent coated with 50 cycles using spherical milling agents and then removing them of certain embodiments disclosed herein.

[0031] FIGS. 5A-5B are representative FlowCam images of a large fraction of agglomerated particles after a sufficient number of coating cycles without milling (FIG. 5A) and principally primary particles after even more coating layers than the control in 5A with initial coating layers performed in the presence of milling agents (FIG. 5B) according to some embodiments of the present disclosure.

[0032] FIG. 6 is a representative graph illustrating particle size distribution of spray-dried formulation coated with a representative number of ALD coating cycles and different milling agents added to the reactor for comparison, as measured by FlowCam (dashed lines) and Mastersizer (solid lines) according to embodiments of the present disclosure.

[0033] FIGS. 7A-7B are representative scanning electron micrograph (SEM) images of spray-dried formulation powder coated with the same number of ALD coating cycles without milling agents (FIG. 7A, control) and spray-dried formulation powder coated in the presence of milling agents for the initial ALD coating cycles (FIG. 7B) according to one embodiment of the present disclosure.

[0034] FIG. 8 is a representative graph illustrating damaged coated particles or microparticles created in absence of milling agents (control) and then demonstrating that agglomeration was dramatically reduced in the presence of milling agents throughout either some or all ALD coating cycles according to one embodiment of the present disclosure.

[0035] FIGS. 9A-9B are schematics illustrating particle agglomeration and / or particle adhesion observed before, during and after coating processes disclosed herein according to certain embodiments of the present disclosure. FIG 9A illustrates how particle agglomeration and / or particle adhesion can cause defective coatings and potentially holes in coated particles due in part to reaction chamber adhesion, or powder bed adhesion of particles referred to as agglomeration according to certain embodiments of the present disclosure. FIG. 9B illustrates anexemplary image of agitated milling agents and agglomerated particle interactions where agglomeration can be reduced by presence of milling agents according to certain embodiments of the present disclosure.DEFINITIONS

[0036] To facilitate an understanding of certain disclosures and concepts herein, the following definitions are provided. Otherwise, all technical terms have their ordinary, art-recognized definitions.

[0037] When introducing elements of the embodiments described herein, the articles “a”, “an”, “the” and “said” are intended to mean that there are one or more of the elements. The terms “comprising”, “including” and “having” are intended to be inclusive and mean that there may be additional elements other than the listed elements.

[0038] Concentrations, amounts, and other numerical data may be expressed or presented herein in a range format. It is to be understood that such a range format is used merely for convenience and brevity and should be interpreted flexibly to include not only the numerical values explicitly recited as the limits of the range, but also to include all the individual numerical values or subranges encompassed within that range as if each numerical value and sub-range is explicitly recited. As an illustration, a numerical range of “about 2 to about 50” should be interpreted to include not only the explicitly recited values of 2 to 50, but also include all individual values and sub-ranges within the indicated range. Thus, included in this numerical range are individual values such as 2, 2.4, 3, 3.7, 4, 5.5, 10, 10.1, 14, 15, 15.98, 20, 20.13, 23, 25.06, 30, 35.1, 38.0, 40, 44, 44.6, 45, 48, and sub-ranges such as from 1-3, from 2-4, from 5-10, from 5-20, from 5-25, from 5-30, from 5-35, from 5-40, from 5-50, from 2-10, from 2-20, from 2-30, from 2-40, from 2-50, etc. This same principle applies to ranges reciting only one numerical value as a minimum or a maximum. Furthermore, such an interpretation should apply regardless of the breadth of the range or the characteristics being described.

[0039] The term “about,” as used herein, can refer to variation of in the numerical quantity that can occur, for example, through typical measuring techniques and equipment, with respect to any quantifiable variable, including, but not limited to, mass, volume, time, distance, and amount. Further, given solid and liquid handling procedures used in the real world, there is certain inadvertent error and variation that is likely through differences in the manufacture, source, or purity of the ingredients used to make the compositions or carry out the methods and the like.The term “about” also encompasses these variations, which can be up to ± 5%, but can also be ± 4%, 3%, 2%, 1%, etc. Whether or not modified by the term “about,” the claims include equivalents to the quantities.

[0040] In this disclosure, “comprises,” “comprising,” “containing,” and “having” and the like can have the meaning ascribed to them in patent laws and can mean “includes,” “including,” and the like, and are generally interpreted to be open ended terms. The terms “consisting of’ or “consists of’ are closed terms, and include only the components, structures, steps, or the like listed in conjunction with such terms, as well as that which is in accordance with patent laws. “Consisting essentially of’ or “consists essentially of’ can have the meaning generally ascribed to them by patent laws. For example, these terms are generally closed terms, with the exception of allowing inclusion of additional items, materials, components, steps, or elements, that do not materially affect the basic and novel characteristics or function of the item(s) used in connection therewith. For example, trace elements present in a composition, but not affecting the composition’s nature or characteristics would be permissible if present under the “consisting essentially of’ language, even though not expressly recited in a list of items following such terminology. In this specification when using an open-ended term, like “comprising” or “including,” it is understood that direct support should be afforded also to “consisting essentially of’ language as well as “consisting of’ language as if stated explicitly and vice versa.

[0041] As used herein, “milling screen” can refer to a barrier, mesh, sieve, or screen with open spaces or apertures to allow the passage of particles smaller than the space or aperture and reduce or prevent passage of molecules or particles larger than the space or aperture. Examples of milling screens can include, but are not limited to a mesh, a wire mesh, plastic mesh, or a screen. A milling screen does not include a gel membrane or other sticky membrane where particles or milling agents would adhere to such a feature. Milling screens disclosed herein can provide flow of particles of sufficient size (e.g., smaller than a pore or opening) to pass through one or more times during a coating process.DETAILED DESCRIPTION

[0042] In the following sections, various exemplary compositions and methods are described to detail various embodiments. It will be obvious to one skilled in the art that practicing the various embodiments does not require the employment of all or even some of the specific details outlined herein, but rather that concentrations, times and other specific details may be modifiedthrough routine experimentation. Tn some embodiments, well known methods or components have not been included in the description.

[0043] Atomic Layer Deposition (ALD) is a vapor-phase process where thin conformal shells or layers of chemical compounds are grown atomic layer by atomic layer on the surfaces of powders using repeated multi-step reaction sequences. In certain embodiments disclosed herein, coating processes conducted using fluidized bed reactors, rotary reactors, and other systems used in the relevant art can be improved by introduction of milling agents into a coating chamber concurrently with the thermostable pharmaceutical-agent containing particles or microparticles to be coated so that all particle or microparticle surface sites are evenly exposed with minimal agglomeration of particles throughout the process and uniform coatings are obtained.

[0044] One issue using these ALD systems is that with certain particle sizes and particle densities, cohesion between particles increases due to either electrostatic interaction, van-der-Waals forces, capillary forces, or hydrogen bonds or other features or any combination of these conditions. Hydrogen bonds or capillary forces can be dominant, for example, during a water dosing cycle of alumina ALD at low deposition temperatures (e.g., less than 100° C). Therefore, with strong cohesive forces, clusters of agglomerated particles can form in the bulk phase of the powders and these clusters are difficult to disperse, even by the forces that the movement within the ALD systems can exert on the particles. As a result, instead of uniform mixing of the powder bed, as observed in ideal coating processes, the coating on the particles or microparticles is uneven and imperfect. The lack of even or uniform mixing during coating processes such as ALD can then result in loss of product and in certain cases, an inferior product produced due to uneven coating of agglomerated particles.

[0045] As the coating process proceeds, the agglomerates both can increase in size as more particles adhere to each other and solidify further as the coating near the contact points of the particles in the agglomerates increases in thickness to eventually form a stable neck. In certain cases, points of contact between the particles cannot be exposed to the chemical precursors to the same extent as accessible surfaces and coatings are thinner at these areas and in some cases form holes. Agglomerates can also intermittently redisperse or break apart after several coating cycles during a coating process due to the mechanical agitation of a reactor bed during coating. When such agglomerates are redispersed either intermittently during the coating process or by handling, grinding, or sieving after coating is complete, contact points can provide weak sectionsof coating that is less protective than the sections of the coatings that were continuously exposed during the ALD process having a more uniform surface. Some areas can even be absent of coatings, depending on the proximity of the contact points, or the fracturing process can lift layers off some particle or microparticle surfaces. These results render such a process imperfect and unideal.

[0046] In some embodiments and further to paragraphs

[0043] -

[0045] above, when microparticles or particles disclosed herein are subjected to a coating process (e.g. ALD), the microparticles or particles can become unevenly coated, adhere to surfaces, and can adhere to one another and, in some instances, form defective particles with thin or no coating and can lead to holes in certain regions causing incomplete or coating and risk loss of therapeutic agent(s) contained therein. See for example, FIG. 9A , a schematic illustrating how agglomerated particles (as illustrated on the left side of the figure) can cause defective coatings in coated particles (as illustrated on the right side of the figure). For example, particles can agglomerate by adhering to each other (e.g., particles adhering to other particles), as illustrated on the left side of FIG. 9A.

[0047] In certain embodiments and further to paragraphs

[0043] -

[0046] above, compositions and methods to improve coating outcome of ALD system coated particles or coated microparticles are disclosed. In accordance with these embodiments, having uniform coatings of difficult-to-fluidize and cohesive powders requires sufficient impact to agglomerated particles or microparticles during a coating process by milling agents or other agents to continuously, or intermittently disperse powders to reduce formation of agglomerates. With decreasing particle sizes and particle density, cohesion between particles typically increases due to one or more of electrostatic interaction, van-der-Waals forces, capillary forces, or hydrogen bonds. Hydrogen bonds and capillary forces can be dominant during the water dosing cycle of alumina ALD at low deposition temperatures (e.g., less than 100 °C), often needed for coating of pharmaceutical particles. Such particles include, but not limited to, spray-dried glassy agents or other formulations in a size range of about a few micrometers often used in pharmaceutical or other applications. Adhesion forces such as hydrogen bonds or capillary forces may be even stronger for the glassy, pharmaceutical agent containing organic particles described herein, especially in the presence of moisture or other ALD chemical as required for the coating process.Conventional devices and methods that are used to aid fluidization, such as low-frequencymechanical vibrations fail to provide sufficient agitation to reduce or prevent agglomeration of strongly cohesive particles. The purge gas used for fluidization purposes does not exert sufficient force to prevent, reduce or remove agglomerated particles, especially at the low pressures and low feed rates often used for ALD system (e.g., ALD) coating processes. Agglomerates of particles or microparticles to be coated can be removed from the coating chamber, manually impacted or sieved to remove and / or reduce the number of agglomerates, and redispersed into the coating chamber intermittently to minimize uneven coatings; however, intermittent deagglomeration of micron-sized agglomerates is intrinsically difficult, this removal-and-reintroduction process is disruptive to the coating process, it is time consuming, it requires stopping the process, and can cause product loss. In addition, maintaining sterility will be difficult in part due to particles exposed to potential contaminants.

[0048] In other embodiments and further to paragraphs

[0043] to

[0047] above, introducing one or more milling agents into a coating chamber of a chemical vapor deposition device concurrently with the pharmaceutical agent containing-particles or microparticles to be coated as disclosed herein in combination with mechanical agitation can reduce and / or prevent agglomeration or increase deagglomeration. In certain embodiments, the pharmaceutical agent containing-particles or microparticles can be dry or essentially dry. In accordance with these embodiment, the dry particles can be formed in the presence of glass-forming agents or other agents capable of making thermostable glassy matrices containing one or more therapeutic agents. In accordance with these embodiments, milling agents, through sheering, mixing, and impacting forces between milling agents and agglomerated therapeutic agent-containing particles or microparticles in presence of mechanical agitation of the coating chamber can deagglomerate agglomerated particles or microparticles or coated or partially coated particles or microparticles within the coating chamber. In other embodiments, one or more milling agents disclosed herein can be composed of different organic or inorganic materials. In certain embodiments, one or more milling agents disclosed herein can be composed of specific inorganic material(s) identified to deliver optimum impact for dislodgement at an optimum time interval. In accordance with these embodiments, the one or more milling agents can be composed of specific material (e.g., inorganic material) that causes minimal damage to the particles or microparticles being coated within the coating chamber.

[0049] In certain embodiments and further to paragraphs

[0043] to

[0048] above, milling agents disclosed herein can be combined and homogenized with pharmaceutical agent containing-particles or microparticles to be coated before introduction to an ALD system or milling agents can be added to ALD systems at the same time, before or after a plurality of pharmaceutical agent containing-particles or microparticles to be coated are introduced to the ALD system. In some embodiments, the pharmaceutical agent containing-particles or microparticles to be coated can be formed by dehydration, lyophilization, vacuum-drying, spray drying, or spray-freeze-drying to form a powder of pharmaceutical agent containing-particles or microparticles prior to introducing them to the ALD system disclosed herein. In some embodiments, the spray-dried powder and milling agent mixture can be violently vortexed or ground to break apart clumps in the spray dried materials. This vortexing or grinding step can be essential for sticky powders that form with actives such as lipid nanoparticles (LNPs) harboring one or more pharmaceutical agent (e.g., polynucleotide) or when spray drying is carried out at low temperatures resulting in wet and clumpy powders (e.g., agglomeration). In certain embodiments, agitation to the coating chamber may not be sufficient to break the sturdy clumps. In some embodiments, powders that appear to flow freely can still need milling agents disclosed herein due to remaining microscopic clumps. In certain embodiments, well dispersed powder and milling agent mixtures in a coating chamber can be dried for a period without any agitation before starting the coating process. Typically, wet and sticky powders can become less sticky when they are drier. Agitating wet and sticky powders before they are dry can cause separation of the milling agents and powder and reforming of larger clumps before the coating starts. In accordance with these embodiments, reformed clumps may not re-disperse during coating instead they can float on top of a powder bed and get coated as a clump which can lead to gaps in coating layers that can lead to pinholes when the particles are dislodged from one another. In other embodiments, composition and methods disclosed herein using milling agents solve this issue with clumping and agglomeration resulting in an improvement of the ALD coating process of pharmaceutical agent-containing particles and microparticles.

[0050] In certain embodiments and further to paragraphs

[0043] to

[0049] above, a coating chamber in the chemical vapor deposition system can include a milling screen. In accordance with these embodiments, the milling screen can be a mesh (e.g., wire, plastic, or other material) or a screen or certain non-sticky permeable barriers having pores of a pre-determined size. Insome embodiments, positioning of the milling screen in a chemical deposition processing chamber or outside a chamber can vary. In some embodiments, the milling screen can be in the middle or closer to the inlet or outlet of the coating chamber. In some embodiments, the mesh can be closer to the outlet of the coating chamber. In these embodiments, milling agents contemplated herein can be introduced on top of the milling screen (e g., wire mesh or screen). In certain embodiments, chemical vapor deposition systems contemplated herein that include a milling screen can be an Atomic Layer Deposition (ALD) system as a vapor-phase process where thin conformal shells of chemical compounds are grown atomic layer by atomic layer on the surfaces of powders using repeated multi-step reaction sequences. In accordance with certain embodiments, fluidized beds can be used to continuously agitate powder particles in a gas stream at reduced or elevated pressure and a mechanical agitation device such as an external sonicator, vibrating motor, impactor, or internal impeller can be used to continuously agitate the milling agents with pharmaceutical agent containing-particles or microparticles within the coating chamber as chemical precursors are added so that all surface sites are evenly exposed throughout a process and uniform coatings are obtained for improved outcome alone or in combination with other devices disclosed herein.

[0051] In certain embodiments and further to paragraph

[0043] to

[0050] above, the compositions and methods described herein provide compositions and method for implementing mechanical forces, including introducing milling agents into a coating chamber in combination with the plurality of pharmaceutical agent containing-particles or microparticles to be coated to agitate (e.g., sheer, mix, impact) the mixture of milling agents and pharmaceutical agent containing-particles or microparticles to be coated inside the coating chamber with sufficient intensity or agitation to overcome particle / particle adhesion forces for micron and sub-micron sized particles that lead to agglomeration due to the presence of the milling agents. In accordance with these embodiments, impact can be obtained with the addition of milling agents into the coating chamber during for example, an ALD cycle in combination with the plurality of pharmaceutical agent-containing particles or microparticles to be coated within the coating chamber and sufficient mechanical agitation such as vibration, impact, or stirring / impelling. In some embodiments, more abrasive milling agents, larger milling agents, or milling agents of different geometric shapes can improve coating uniformity with reduced agglomeration. For example, using larger sized milling agent when compared to the size of the plurality of thetherapeutic agent-containing particles or microparticles to be coated within the coating chamber can be advantageous. In certain embodiments, milling agents can be about 1.1 to about 5 or 10 times larger than the particles to be coated. In certain embodiments, milling agents can be about 1.1 to about 5 times larger than the particles to be coated. In some embodiments, the milling agents can be about 1.1 to about 4 times larger than the particles to be coated. In some embodiments, the milling agents can be 1.1 to about 5 times heavier or denser than particles to be coated. In some embodiments, the milling agents can be 1.5 to about 3.5 times heavier or denser than particles to be coated. In other embodiments, less abrasive milling agents with lower or similar density to an average density of uncoated and / or partially coated (therapeutic agentcontaining particles or microparticles can be less effective in reducing or preventing agglomeration of particles before, during and / or after initial coating processes (e.g., up to about 30 coating cycles or more). In some embodiments, milling agents can remain in a powder bed throughout the coating process. In some embodiments, less dense (e.g., having about the same or a lower density to an average density of uncoated and / or partially coated therapeutic agentcontaining particles or microparticles) and / or less abrasive milling agents can remain in a powder bed after initial coating of therapeutic agent-containing particles or microparticles (e.g., about 1 to about 40 or more, about 1 to about 30, about 20 coating layers) or remain throughout a coating process because these less abrasive milling agents can be less damaging during a particular coating process of therapeutic agent-containing particles or microparticles and / or easily separated from an end product. In some embodiments, larger or more abrasive milling agents can be present during initial coating cycles where cohesive surface properties of the glassy pharmaceutical agent containing particles causes the most or more agglomeration, but the larger or more abrasive milling agents can be replaced or substituted for smaller, less dense, or less abrasive milling agents after initial coating layers (e.g., about 5, about 10 or about 20 layers or more) and these less dense milling agents can be maintained for several additional coatings or until a final coating layer is applied to particles contemplated herein, if desired. In accordance with these embodiments, this process of using large more dense milling agents and then less abrasive or less dense milling agents than a first large more dense milling agent can reduce adhesion, reduce or prevent agglomeration as well as reduce or prevent potential reagglomeration of particles being coated with reduced damaging effects on coating layers, leading to more uniform coating layers for completely encasing therapeutic agents within the coatedparticles. In some embodiments, a less abrasive or less dense milling agent can include, but is not limited to, a polymer powder having a density closer to the particles or even less than the particles being coated (e.g., a polymer powder of about 1.0-1.5 g / cm3) may work. The current media with their density of 4.5 to 6 g / cm3 and their relative large size are like big heavy bowling balls hitting the little particles with their egg shell. Not hard enough to crush them since the images show them to be intact as spheres, but hard enough to damage the egg shell beyond repair. In accordance with these embodiments, these processes can lead to a more predictable end product concentration of use for more accurate timed and delivered therapeutic agent to a subject at reduced cost and improved production.

[0052] Other embodiments and further to paragraphs

[0043] -

[0051] above, provide for novel methods for maintaining microparticle distribution and / or reducing or preventing microparticle agglomeration within the coating chamber of the system such as atomic layer deposition (ALD) system and / or improving delivery or syringability of coated microparticle compositions or formulations contemplated herein. In certain embodiments, ALD systems disclosed herein further include milling agents for additional agitation. In accordance with these embodiments, methods disclosed herein utilize milling agents that can include any inorganic or organic material 1.1-100, 2-50, 2-25, 2-15 or 2-10 or 1.1 to 5 times larger than the size of a primary or average size particle or microparticle to be coated within a coating system (e.g., ALD). For example, the milling agents can be from about 1.5 to about 95, about 2 to about 90, about 2 to about 85, about 2 to about 80, about 2 to about 75, about 2 to about 70, about 2 to about 65, about 2 to about 60, about 2 to about 55, about 2 to about 50, about 2 to about 45, about 2 to about 40, about 2 to about 35, about 2 to about 30, about 2 to about 25, about 2 to about 20, about 2 to about 15, or about 2 to about 10 times larger than the size of the primary particle or microparticle to be coated within the system. In other embodiments, the milling agents within the coating chamber can further include at least a second agent including an inorganic or organic material for disrupting agglomeration or adherence of coated particle or microparticles in a coating chamber disclosed herein. In certain embodiments, a second inorganic or organic agent can be a different size than a first inorganic or organic material. In other embodiments, the milling agents disclosed herein can include a combination of inorganic or organic materials, including but not limited to glass, ZrCh, ZrCh / SiCh, Fe, or AI2O3 or other suitable inorganic or organic material. In some embodiments, the milling agents disclosed herein can include a combination of one or more compositions ofinorganic material, one or more sizes or shapes of inorganic material, and / or one or more compositions of inorganic material of one or more sizes of inorganic material.

[0053] In certain embodiments and further to paragraphs

[0043] -

[0052] above, the milling agents added into the coating chamber in combination with the particles or microparticles to be coated can further include highly dense materials (e.g., ZrCh-based particles), which can be less susceptible to fracture while allowing adequate sheering, mixing, or impact in a coating chamber to reduce or eliminate agglomeration of a plurality of microparticles or particles or partially coated microparticles or particles (e.g. up to 30 coating layers or more). In certain embodiments, a highly dense material can include, but is not limited to, glass, ZrCh, ZrCh / SiCh, Fe, or AI2O3 or combination thereof or mixture thereof. In other embodiments, extremely hard materials can be avoided to reduce grind of walls or damage or scratches of certain types of coating chambers. For example, extremely hard materials can grind walls of a stainless-steel reactor and create divots or dents or scratches which can alter a coating process and affect flow of the particles during coating.

[0054] In other embodiments and further to paragraphs

[0043] -

[0053] above, milling agents with similar chemistry to a final product coating can aid in preventing introduction of potentially new materials to the product while allowing adequate sheering, mixing, or impact to the coating chamber to reduce or eliminate agglomeration of the plurality of microparticles or particles or coated microparticles or particles. In certain embodiments, milling agents with similar chemistry to the final product can include, but is not limited to, media containing alumina or sugars. In other embodiments milling agents with similar chemistry to the reactor components can include, but is not limited to, glass or stainless steel.

[0055] In certain embodiments and further to paragraphs

[0043] -

[0054] above, an improved method for coating microparticles or particles is disclosed having improved features when compared to traditional coating processes as well as when compared to coating by a coating chamber with only a sonicator, impactor, or vibrator attached thereto. In accordance with these embodiments, an ALD system for coating microparticles can be used and can include milling agents within a coating chamber having a plurality of particles or microparticles being coated or to be coated. In other embodiments, these ALD systems can include a sonicator, impactor, and / or vibrator attached to the exterior of the ALD system and / or internal stirrer or impeller for generating coated particles or microparticles having reduced or eliminated adherence to otherparticles or microparticles, the coating chamber, and / or filter. In certain embodiments, a coating or reaction chamber can define a cylindrical tube, cone, cube or other configuration of the coating chamber. In other embodiments, the coating chamber can further include an inlet at a first end of the coating chamber; an outlet at a second end of the coating chamber; and one or more walls, nodules, or surfaces within the coating chamber. The coating chamber can further include a sonicator, impactor, vibrator, and / or internal stirrer or impeller associated thereto for generating coated particles or microparticles having reduced or eliminated adherence to other particles or microparticles, the coating chamber being attached to the exterior of the reactor chamber. In accordance with these embodiment, the coating chamber can be configured to allow flow of process gases or purge gas (e.g., argon, TMA vapor, water vapor, mixtures of argon with TMA vapor, or mixtures of argon with water vapor) through the coating chamber while retaining the plurality of particles or microparticles within the coating chamber between the first end of the coating chamber and the second end of the coating chamber, the interior surfaces defining an internal volume, where the coating chamber is configured to receive the plurality of particles or microparticles; and milling agents as disclosed herein for preventing or reducing agglomeration or adherence of the plurality of particles and / or microparticles and coated microparticles within the coating chamber. In some embodiments, the coating chamber can be a fluidized bed reactor. In other embodiments, the coating chamber can be a rotary reactor.

[0056] In some embodiments and further to paragraphs

[0043] -

[0055] above, the milling agents are added to the coating chamber to prevent or reduce adherence of particles or microparticles or coated microparticles to each other during the coating process to prevent formation of coated agglomerates, thereby improving delivery or syringability of coated microparticle compositions or formulations contemplated herein. In certain embodiments, the milling agents impact the particles and / or microparticles within the coating chamber during the coating process by sheering, mixing, and impacting the particles and / or microparticles interactions with one another to reduce agglomeration of the particles or microparticles within the coating chamber. In accordance with these embodiments, sheering, mixing, and impacting of the particles and / or microparticles within the coating chamber by the milling agents contemplated herein reduce or eliminate the plurality of particles or microparticles from agglomerating to one another within the coating chamber. These interactions, can promote recovery of higher percentage or portion ofdeagglomerated coated material product compared to traditional methods of coating particles or microparticles without the use of milling agents.

[0057] In certain embodiments and further to paragraphs

[0043] -

[0056] above, milling agents impact coating of the therapeutic agent-containing particles or microparticles during one or more coating cycles of the microparticles or particles within a coating chamber by reducing direct particle to particle or microparticle to microparticle interaction. In some embodiments, the coating process can be interrupted (e.g. after up to 20 cycles or after up to 30 cycles or more), and the milling agents are removed or separated from coated particles or microparticles, for example by sieving, magnetic forces, aerosolized separation, or other appropriate separation methods. In certain embodiments, these deagglomerated or agglomeration-free coated particles can be re-introduced into the coating chamber, and the coating process is then re-started without the presence of the milling agents in the coating chamber. In certain embodiments, the milling agents can be present during only one round of a coating process or over the course of numerous rounds of coating. In other embodiments, inclusion of milling agents in multiple coating rounds can depend on the characteristics of the material to be coated such as the number of microparticles or particles to be coated or the size of the microparticles or particles to be coated. In some embodiments, agitation by milling agents as disclosed herein could cause damage or breakage of the coating(s) deposited onto the particles or microparticles to be coated and therefore is limited in duration and controlled to provide effective deagglomeration and removal from the walls while minimizing damage to the particles or microparticles, or coated particles or microparticles. In some embodiments, deagglomerated coated particles can have some damaged layers. In accordance with these embodiments, damaged layers can be further coated with or without the plurality of milling agents to sequester or fdl defects of a damaged layer to attain a defect-free coating. In some embodiments, the coated particles (e.g. about 10 coating cycles or more) have little or no agglomeration because exposed particle surfaces are coated. In certain embodiments, it is contemplated that agglomeration appears in the early cycles such as less than 10 or less than 20 or less than 30 coating cycles and then there is little or no further agglomeration. In accordance with these embodiments, milling agents can be used only in these early cycles and then removed for further coating of particles or microparticles, if desired.

[0058] In certain embodiments and further to paragraphs

[0043] -

[0057] above, methods are provided for creating improved coated therapeutic agent-containing microparticles containingtargeted antigens, thermostable agents, thermal stable chemicals such as pharmaceutical agent, and thermal stable immunogenic formulations. In certain embodiments, methods disclosed herein for creating improved coated microparticles or particles from thermostable agents include introducing milling agents into the coating chamber in combination with particles and / or microparticles to be coated for the purpose of sheering, mixing, or impacting the particles and / or microparticles to be coated for creating more uniformly coated glassy particles containing at least one antigen with reduced or eliminated agglomeration or microparticle adherence.

[0059] In certain embodiments and further to paragraphs

[0043] -

[0058] above, microparticles, particles, thermostable microparticles, particles or powders containing microparticles or particles having one or more therapeutic agent contained therein can be introduced to a coating chamber or reaction chamber in addition to introducing milling agents to the reaction or coating chamber as contemplated herein that are not or never removed from the coating chamber and reintroduced to the coating chamber for additional coatings without milling agents present (e.g., the milling agents are not removed until completion of coating). In other embodiments, only the milling agents can be removed from the coating chamber to then allow additional coatings of the microparticles or particles or thermostable microparticles or particles thereof or powders containing microparticles or particles. In certain embodiments disclosed herein, the coating chamber is devoid of a sieve for repeated, intermittent use as a deagglomeration device during the coating process.

[0060] In other embodiments and further to paragraphs

[0043] -

[0059] above, a removable sieve can be used to sieve particles before introduction to the coating chamber or prior to coating to remove agglomerates that may be present in the starting material or used after completion of a coating process. In other embodiments, a removable sieve can be used at any point of the coating process to remove the milling agents, leaving the microparticles or particles or thermostable microparticles or particles thereof or powders containing microparticles or particles for additional coatings to be completed without the milling agents present. In some embodiments, a final quality control sieving process can also be facilitated using brief exposure to suitable drying agents to minimize the cohesive properties of coated particle or microparticle products.

[0061] In certain embodiments and further to paragraphs

[0043] -

[0060] above, thermal stable chemicals or agents or antigens contemplated of use for coating in ALD systems disclosed herein can include, but are not limited to, any pharmaceutical agent or biologic or chemical agentcapable of being coated by ALD or other coating process. In some embodiments, one or more antigens and / or therapeutic agents can initially be embedded in an organic glassy matrix or a glass forming agent or thermostable or thermostabilized or to form a shell stabilizing the antigens or therapeutic agent prior to introduction to an ALD system disclosed herein for coating. In accordance with these embodiments, compositions and methods known in the art can be used to create stabilized or thermostable particles prior to coating. In other embodiments, at least a primary and a boost dose of the same antigens or same therapeutic can be encased in coated particles or microparticles disclosed herein separated by coating layers where the particles or microparticles have reduced agglomeration or adhesion using devices disclosed herein. In other embodiments, two or more different antigens or therapeutic agents or compounds can be dispersed in a single microparticle or particle in the same or different layers (e.g. separated by coating layers) or multiple particles or microparticles each containing one or more antigen or therapeutic agent that can be mixed in a single composition and then prepared for coating by techniques disclosed herein. In other embodiments, antigen or therapeutic agent-containing particles or microparticles or powders disclosed herein can include immunogenic agents against two or more pathogens either in the same or in separate particles or microparticles.

[0062] In other embodiments and further to paragraphs

[0043] -

[0061] above, antigen-, therapeutic agent- or immunogenic agent-containing particles or microparticles disclosed herein can have a central or innermost antigen-, therapeutic agent- or immunogenic agent-containing microparticle or particle including at least one immunogenic agent, therapeutic agent or antigen and optionally, at least one glass-forming agent; and one or more coating layers using methods disclosed herein for covering or encasing the central or innermost antigen-, therapeutic agent- or immunogenic agent-containing microparticle or particle with reduced adherence of the microparticles or particles to one another. In some embodiments, a primary antigen-, therapeutic agent- or immunogenic agent-containing microparticle can be created by dehydration, lyophilization, vacuum-drying (and optionally, milling of solid residues), spray drying, other evaporative drying technique, or spray-freeze-drying prior to introducing to an ALD system disclosed herein. In accordance with these embodiments, one, two, three, four, five, up to 20, up to 20, up to 100, up to 250 or more coating layers deposited during the ALD cycles can encase the thermostable microparticles or particles where the coating layers are readily dissolvable in a subject once administered, to expose the antigen, therapeutic agent or immunogenic agent of thecoated particles or microparticles to the subject. Tn certain embodiments, additives and methods disclosed herein provide for production of more uniformly coated particles with reduced loss and reduced side effects of adherence and agglomeration leading to an increase in production, a more reliable end-product and reduce costs in production.

[0063] In other embodiments and further to paragraphs

[0043] -

[0062] above, methods are provided for conducting improved particle or microparticle coatings within an atomic layer deposition or chemical deposition system (e.g., ALD system) having a sonicator, impactor, or vibrator, and / or internal stirrer or impeller by introducing one or more milling agents in combination, mixed or sequentially added with the particles or microparticles to a reaction chamber of a deposition system for coating as disclosed herein for generating coated microparticles or particles having reduced or eliminated agglomeration. In accordance with these embodiments, the one or more milling agents disclosed herein for generating coated microparticles having reduced or eliminated agglomeration provides improved uniformity of coated particles, reducing issues of imperfections of the coated particles such as holes, cracks or incomplete coating of one or more coating layers on the microparticle for increased productivity with improved and more reliable production of product. In other embodiments, the one or more milling agents are added directly into a coating chamber disclosed herein which can result in reduced adherence and / or agglomeration by at least 1.0% up to 100% compared to systems without one or more milling agents added directly into a coating chamber. For example, reduced adherence and / or agglomeration can range from about 1.0% up to about 99.9%, 1.0% up to about 95%, about 1.0% up to about 90%, about 1.0% up to about 85%, about 1.0% up to about 80%, about 1.0% up to about 75%, about 1.0% up to about 70%, about 1.0% up to about 65%, about 1.0% up to about 60%, about 1.0% up to about 55%, about 1.0% up to about 50%, about 1.0% up to about 45%, about 1.0% up to about 40%, about 1.0% up to about 35%, about 1.0% up to about 30%, about 1.0% up to about 25%, about 1.0% up to about 20%, about 1.0% up to about 15%, about 1.0% up to about 10%, compared to systems without one or more milling agents added directly into a coating chamber or without contact with a milling agent as disclosed herein.

[0064] In other embodiments and further to paragraphs

[0043] -

[0063] above, at least one immunogenic agent or antigen or other therapeutic agent can include, but is not limited to, one or more of a polypeptide or fragment thereof, a polynucleotide, a pharmaceutical agent or chemical, a whole organism or derivative or polypeptide derived therefrom or small molecule or antibodyor fragment thereof, or a combination thereof. Tn accordance with these embodiments, the at least one immunogenic agent or antigen or other therapeutic agent includes but is not limited to one or more of a viral antigen, a bacterial antigen, a toxin, a fungal agent or other pathogenic agent, a pharmaceutical agent (e.g., anti-cancer, anti-inflammatory or other agent) or a combination thereof or small molecule or complex compound. In some embodiments, the at least one immunogenic agent or other therapeutic agent can also include, but is not limited to, a recombinant peptide, a recombinant protein, a peptide derived from a target protein or pathogen, a synthetic peptide or protein, a virus-like particle, a live virus, a live, attenuated virus, an inactivated virus, a bacterial antigen, a bacteriophage or phage, a fungus, a prion; a fragment thereof, a derivative thereof, or a combination thereof.

[0065] In some embodiments and further to paragraphs

[0043] -

[0064] above, each layer of the one or more coating layers can include at least one metal agent (e.g. metal oxide or metal alkoxide) or other suitable agent. In certain embodiments, the one or more coating layers can include at least one of an organic material, a metallo-organic material, metal oxides, metal alkoxides, and / or aluminum-based coating layer. In accordance with these embodiments, one or more coating layers can include, but is not limited to, one or more of aluminum oxide (A12O3), an aluminum alkoxide (e.g. alucone), silicon dioxide (SiCh), Zinc dioxide (ZnCh), or zinc oxide (ZnO), titanium dioxide (TiCh), and silicon nitride (Si3N4) or combinations thereof or alternating layers thereof. In some embodiments, one or more coating layers can include, but are not limited to, one or more of aluminum oxide (AI2O3), an aluminum alkoxide (e.g., alucone), silicon dioxide (SiCh), titanium dioxide (TiCh), or zinc oxide (ZnO), alone or in a suitable combination composition or alternating layers thereof.

[0066] In certain embodiments and further to paragraphs

[0043] -

[0065] above, processes and compositions for producing coated therapeutic agent-, antigen-, immunogen-, compoundcontaining particles or microparticles disclosed herein use less therapeutic agent, antigen, immunogen or compound than used to formulate current pharmaceutical such as vaccines and other active agents (e.g., cost saving, active-agent sparing), and provide enhanced efficacy after a single administration with reduced cost for production and increased reliability and uniformity of compositions and further have reduced adherence or agglomeration further improving production of coated particles for time-release of active agents contemplated herein. In other embodiments, therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticlesprovide for thermostable formulations that eliminate and / or reduce refrigeration requirements (e.g., cold chain refrigeration requirements), limit the concentrations of adverse agents (e.g., aluminum) administered to subjects, and increase compatibility.

[0067] Some embodiments disclosed herein and further to paragraphs

[0043] -

[0066] above, relate to methods of dehydration and formulation parameters, where these parameters can be adjusted in order to control nucleation rates, glass transition temperatures, and other material properties of the therapeutic agent-, antigen-, immunogen-, and / or compound-containing particles or microparticles. In accordance with these embodiments, these particles or microparticles described herein are thermostable to tolerate the coating process with little to no degradation or loss of therapeutically active material. Using the coating method modifications disclosed herein, the resulting coated particles or microparticles have reduced adherence or agglomeration during the coating process.

[0068] In certain embodiments and further to paragraphs

[0043] -

[0067] above, pathogenic viruses or fragments derived thereof or modified viruses are contemplated of use in coated particles or microparticles disclosed herein. In accordance with these embodiments, the pathogenic viruses can include, but are not limited to, Ebola viruses or any filo viruses, a papovavirus (e.g., papillomaviruses, including human papilloma virus (HPV)), a herpesvirus (e.g., herpes simplex virus, varicella-zoster virus, bovine herpesvirus- 1, cytomegalovirus), a poxvirus (e.g., smallpox virus), a reovirus (e.g., rotavirus), a parvovirus (e.g., parvovirus B19, canine parvovirus), a picornavirus (e.g., poliovirus, hepatitis A), a togavirus (e.g., rubella virus, alphaviruses such as Chikungunya virus), a hepadnavirus (e.g., hepatitis B virus), a flavivirus (e.g., dengue virus, hepatitis C virus, West Nile virus, yellow fever virus, Zika virus, other alphaviruses or flaviviruses, Japanese encephalitis virus), an orthomyxovirus (e.g., influenza A virus, influenza B virus, influenza C virus), a paramyxovirus (e.g., measles virus, mumps virus, respiratory syncytial virus, canine distemper virus, parainfluenza viruses), rickettsia, a rhabdovirus (e.g., rabies virus), a filovirus (e.g., Ebola virus), or a coronavirus or combinations thereof.

[0069] In some embodiments and further to paragraphs

[0043] -

[0067] above, an agent or antigen can include at least one bacteriophage or other similar agent. In accordance with these embodiments, a bacteriophage can be formulated in coated particles as disclosed herein. In otherembodiments, one or more coated bacteriophages can be used to treat or prevent an infection or multi-drug-resistant infection caused by one or more bacteria.

[0070] In other embodiments and further to paragraphs

[0043] -

[0067] above, at least one pathogenic agent can form part of a particle or microparticle disclosed herein or a polynucleotide or polypeptide derived therefrom. For example, these pathogenic agents or polynucleotides or polypeptides derived therefrom can include, but are not limited to, a fungus, a prion, a bacterium or a toxin of a bacterium, including but not limited to, Pasteurella haemolytica, Clostridium difficile, Clostridium haemolyticum, Clostridium tetani, Corynebacterium diphtheria, Neorickettsia resticii, Streptococcus equi, Streptococcus pneumoniae, Salmonella spp., Chlamydia trachomatis, Bacillus anthracis, Yersinia spp., and Clostridium botulinum or combinations thereof. In yet other embodiments, the pathogenic agent contained in a microparticle can be a toxin, such as ricin toxin or botulinum toxin, a polynucleotide, or polypeptide derived therefrom.

[0071] In some embodiments and further to paragraphs

[0043] -

[0070] above, the at least one pathogenic agent can be contained in a particle or microparticle disclosed herein or a polynucleotide or polypeptide derived therefrom. For example, these antigens can include, but are not limited to Cryptococcus spp. (e.g., neoformans and gatti), Aspergillus spp. (e.g., fumigatus), Blastomyces spp. (e.g., dermatitidis), Candida albicans, Paracoccidioides spp. (e.g., brasiliensis), Sporothrix spp. (e.g., schenkii and brasiliensis), Histoplasma capsulatum, Pneumocystis jirovecii and Coccidioides immitis, or combinations thereof.

[0072] In certain embodiments and further to paragraphs

[0043] -

[0071] above, compositions and methods can be applied to pharmaceutical compositions other than antigen or agent compositions disclosed herein. For example, small molecule drugs (e.g., anti-cancer agents), polynucleotide or siRNAs or mRNAs, DNA, chimeras or chimeric polynucleotides or carbohydrates or other agents and biologies can be similarly coated as disclosed for immunogenic agent-containing glassy microparticles described herein. As disclosed herein, the coating layers can provide for a level of temporally controlled release desirable with certain pharmaceutical agents. The coating layers can serve to reduce exposure to moisture, reducing degradation. These coatings can function to protect water-soluble drug formulations or other moisture sensitive agents from degradation or dissolution until desired exposure to a subject after administration. Further, the embodiments can be used in applications outside of therapeutics. For example, coating layers canbe applied to diagnostic markers. The coating layers can allow delayed release of the marker, allowing sufficient trafficking / uptake time. This delayed release of the marker can be beneficial where the marker has a limited half-life. In certain embodiments, immunogenic compositions disclosed herein or encapsulated small molecules using layering / coating technologies described herein can be administered directly to an affected location of a subject such as the liver or kidney or brain depending on the ability of the deposited composition to remain in the targeted region.

[0073] In some embodiments and further to paragraphs

[0043] -

[0072] above, coating of particles or microparticles having improved uniformity, reduced agglomeration and reduced adherence under coating conditions of an ALD system can be due to the methods or processes disclose herein by including milling agents within an ALD reaction or coating chamber. In accordance with these embodiments, these methods can be used in the manufacture or scale-up manufacture of one or more therapeutic agent-, antigen-, immunogen-, compound-containing particle composition of use to treat, reduce or prevent a health condition in a subject. Subjects that can benefit from such processes and treatments include humans or other mammals or animals. In accordance with these embodiments, a subject can be a fetus, infant, toddler, child, adolescent, mature adult or older adult human or other subject. In certain embodiments, the subject can be a dog (canine), a cat (feline), a horse (equine), cattle (bovine), a goat (hircine), a sheep (caprine), a pig (porcine) or poultry (e.g., chicken, turkey, duck, goose), or other bird, reptile, fish, or another animal.

[0074] In certain embodiments and further to paragraphs

[0043] -

[0073] above, therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles described herein can include a single agent dose or two or more doses of a particular agent or different agents (e.g., prime and boost doses or combination agent formulations). In some embodiments, particles or microparticles can include doses for two or more different agents. In yet other embodiments, agent-containing particles or microparticles including doses of different agents can be combined into a mixture of agent-containing particles prior to coating or introduced on an layer for additional coating or as an layer on a coated microparticle. In certain embodiments, a mixture of agent-containing microparticles can be combined into a single administration for a reduced number of vaccine administrations.

[0075] In some embodiments and further to paragraphs

[0043] -

[0074] above, methods disclosed herein can concern controlled, ultra-rapid freezing rates and agitated coating processes. Inaccordance with these embodiments, therapeutic agent-, antigen-, immunogen-, compoundcontaining microparticles or particles can include at least one glass forming agent, including, but not limited to, trehalose, sucrose, fi coll, dextran, sucrose, maltotriose, lactose, mannitol, hydroxyethyl starch (HES), glycine, cyclodextrin, povidone, or the like. In other embodiments, therapeutic agent-, antigen-, immunogen-, compound-containing microparticles or particles can include at least one glass forming agent and a short peptide (e.g., shell-forming agent such as trileucine or equivalent agent thereof). In certain embodiments, the at least one glass-forming agent can include trehalose. In accordance with these embodiments, these agents can be used to generate glass-like matrices upon freeze-drying. In certain embodiments, when the glass-forming agents are dried during a dehydration process (e.g., spray-drying) in the presence of one or more agents, these form powders (glassy microparticles), containing embedded antigens, agents, and / or compounds. In this dehydrated state which is essentially dry, protein physical and chemical degradation pathways, which require molecular motion, can be inhibited, as are other degradation pathways, stabilizing the antigen, agent, and / or compound in preparation for a coating process disclosed herein (ALD-coating).

[0076] In some embodiments and further to paragraphs

[0043] -

[0075] above, buffers of use for formulations and compositions disclosed herein for storage and / or delivery of a pharmaceutical composition generated by compositions, methods and systems disclosed herein can include, but are not limited to, acetate, succinate, citrate, prolamine, histidine, borate, carbonate or phosphate buffer, or other suitable stabilizing buffer known in the art or a combination thereof. In certain embodiments, a buffer can include histidine, for example, histidine- HC1. In some embodiments, polysaccharides and other agents of use to stabilize agents, antigens or compounds in preparation for coating by a device disclosed herein can include one or more of trehalose, sucrose, ficoll, dextran, sucrose, maltotriose, lactose, mannitol, hydroxyethyl starch, glycine, cyclodextrin, and povidone, or combinations thereof. In certain embodiments, the polysaccharide can be trehalose. In other embodiments, the polysaccharide concentration can be present in a weight-to-volume (w / v) concentration from about 0.1% to about 40% in a composition prior to dehydration or spray-drying. For example, the polysaccharide concentration present range from about 0.1 to about 35%, about 0.1 to about 30%, about 0.1 to about 25%, about 0.1 to about 20%, about 0.1 to about 15%, about 0.1 to about 10%, or about 0.1 to about 5% w / v in the composition prior to dehydration or spray-drying. In another embodiment, the polysaccharide can be a concentrationfrom about 8% to about 11%; or about 9.5% w / v in the agent-, antigen- or compound-containing composition prior to dehydration or spray-drying.

[0077] In certain embodiments and further to paragraphs

[0042] -

[0076] above, a smoothing excipient of use in compositions and methods disclosed herein can be included in the composition to be lyophilized or spray-dried or evaporatively dried prior to coating of the therapeutic agent-, antigen-, immunogen-, compound-containing microparticles or particles can include at least one glass forming agent, particles or microparticles. In accordance with compositions disclosed herein, the smoothing excipient can aid in creation of a smooth(er) agentcontaining particle or microparticle’s surface, which in turn can create improved ability to deposit one or more covering layers on the particle or microparticle. In accordance with these embodiments, having a smooth(er) agent-containing glassy microparticle with reduced inconsistencies on the surface reduces the risk of cracking. In certain embodiments, coating layers described herein, each of which can be about 0.1 nm or thicker, can crack or incompletely cover the agent-containing particle or microparticle due to inconsistencies occurring on the surface of the underlying particle or microparticle creating raised or indented surfaces. In certain embodiments, the smoothing excipient can also function as a stabilizing agent. In some embodiments, the smoothing excipient can be hydroxyethyl starch or another pharmacologically acceptable plasma expander including, but not limited to, serum albumin, human serum albumin, dextran, hetastarch, and plasma protein factor, or the like or a combination thereof. In other embodiments, the smoothing excipient can be hydroxyethyl starch. In some embodiments, the smoothing excipient can be present in a weight-to-volume (w / v) concentration from about 0.1% to about 40% in a composition prior to dehydration or spray-drying. In some embodiments when the smoothing excipient is the same or different from the other stabilizing agent or polysaccharide, the smoothing excipient concentration is from about 0.1% to about 5%; about 0.1% to about 2.5%; about 0.1% to about 1.0%, about 0.1% to about 0.5%, or about 0.1% to about 0.25% in a composition prior to dehydration or spray-drying.

[0078] In certain embodiments and further to paragraphs

[0042] -

[0077] above, agents used in the thermostable agent-containing particles or microparticles of the present disclosure can be of use for prophylactic and / or therapeutic compositions. Suitability of agents for use in therapeutic agent-, antigen-, immunogen-, compound-containing microparticles or particles can be tested by reaction with antibodies or monoclonal antibodies which react or recognize conformationalepitopes present on the intact target of the agent and based on the agent’s ability to elicit the production of neutralizing antiserum. Suitable assays for determining whether neutralizing antibodies are produced are known to those of skill in the art. In this manner, in certain embodiments, it can be verified whether the immunogenic agents of the present disclosure will elicit production of neutralizing antibodies.

[0079] In some embodiment and further to paragraphs

[0043] -

[0078] above, methods disclosed herein having one or more types of milling agents within the coating chamber for at least reducing agglomeration to provide for more uniform syringability and uniform consistency of the compositions for a more reliable and predictable delivery and dosing of an agent, antigen or immunogenic agent disclosed herein. For example, agitation of the reactor in the presence of one or more types of milling agents within the coating chamber along with the microparticles or particles or thermostable microparticles or particles thereof or powders containing microparticles or particles to be coated can produce uniformly coated microparticles or particles for a more uniform composition with reduced loss from clumping between coated particles or coated microparticles.

[0080] In some embodiments and further to paragraphs

[0043] -

[0079] above, ALD or other chemical deposition or coating systems can be used to apply nanometer-thick coatings of inorganic, organic, or metallo-organic materials on the surface of therapeutic agent-, antigen-, immunogen-, and / or compound-containing particles or microparticles with improved reliability due to intermittent or continuous agitation within the coating chamber. In certain embodiments, the coating or sequestering layer can be a metal oxide or metal alkoxide or for example, an aluminum-based material including, for example, an aluminum oxide or an aluminum alkoxide (e.g., alucone). In accordance with these embodiments, the metal oxide or metal alkoxide (e.g., aluminum-containing material) is deposited on or applied to the surface of the plurality of therapeutic agent-, antigen-, immunogen-, and / or compound-containing particle or microparticle to coat the plurality of particles or microparticles in 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, up to 20, up to 50, up to 100, up to 200, up to 300, up to 400, up to 500, up to 600, up to 1,000, up to 1500, up to 2000 or more layers of a metal agent (e.g. metal oxide or metal alkoxide) or non-metal agent coating such as metal oxide- or metal alkoxide- (e.g., aluminum oxide) containing material to form encased particles or microparticles.

[0081] In certain embodiments and further to paragraphs

[0043] -

[0080] above, a binary, ternary, or multi-step reaction sequence can be used to deposit one or more layers of metal oxide (e.g., alumina) or metal alkoxide (e.g., alucone) or other suitable chemical agent on a therapeutic agent-, antigen-, immunogen-, compound-containing particle or microparticle. Therapeutic agent-, antigen-, immunogen-, compound-containing -containing particles or microparticles can be exposed to alternating gas streams containing either vaporized reactive metal compounds (e.g., trimethyl aluminum or titanium chloride) or an oxidizer such as water vapor, ozone, or hydrogen peroxide. In certain embodiments, the number of cycles can be varied to control formation of the coating layer on a plurality of particles or microparticles. In accordance with these embodiments, the one or more milling agents can be added to the coating chamber to maintain consistent, more uniform coating layer coverage of the particles or microparticles with reduced agglomeration of the particles or microparticles being uniformly coated by at least 5.0% or more, or 10.0% or more as compared to particles coated not in the presence of milling agents. In other embodiments, the milling agents can be removed from the coating chamber while the coated particles and / or microparticles remain in the coating chamber for addition coating layers as needed (e.g., using a magnet or other device or process to remove the milling agents.

[0082] In accordance with these embodiments and further to paragraphs

[0043] -

[0081] above, some advantages of depositing one or more coating layers on therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles with improved uniformity include, but are not limited to, that the coating layers can dissolve slowly or at a more accurate coordinated pre-determined rate when the therapeutic agent-, antigen-, immunogen-, compoundcontaining particles are administered to a subject, thus allowing temporal control of the release of the particle contents (e.g., the one or more antigens, therapeutic agents, immunogens or other compounds). Release times can be tailored by adjusting composition(s) of the coating layers and number of and / or thickness of molecular layers applied to the agent-containing particles or microparticles. In some embodiments, up to about 5, up to about 100, up to about 150, up to about 200, up to about 300, up to about 400 or more, up to about 1,000 or more coating layers can be used to form the coated agent-containing particles microparticles of the present disclosure. In certain embodiments, the addition of milling agents during a coating process disclosed herein can lead to a need of fewer coating layers due to uniformity of the coating of the particles and reduced agglomeration. In some embodiments, release of the agents from thecoated therapeutic agent-, antigen-, immunogen-, compound-containing particle’s or microparticle’s layers or core can occur within hours, to about 1 day, or about 7 days or about 30 days or about 60 days or about 90 days or about 120 days or about 180 days, or about 6 months, or about a year, or other predetermined timing after administration to the subject. In some embodiments, release of the coated therapeutic agent-, antigen, immunogen, and / or compound from the particle can occur from up to about 10 days up to / and about 90 days up to / and about 120 up to / and about 150 up to / and about 180, up to / and about 210 or more days after administration to the subject depending on the number of coating layers, material of the coating layers and formulation contained within the coated particles (e.g. therapeutic agent). In some embodiments, release of the innermost agents can occur from about 10 days to about 365 days after administration to the subject. In some embodiments, release of the innermost agents can occur from about 14 days to about 180 days after administration to the subject. Further, in some embodiments, release of the innermost agents can occur from about 18 days to about 90 days after administration to the subject.

[0083] In certain embodiments and further to paragraphs

[0043] -

[0082] above, particle or microparticle size of the encased or fully coated plurality of therapeutic agent-, antigen-, immunogen-, compound-containing or combination therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles can be from about 0.001 pm to about 150.0 pm, about 0.05 pm to about 150.0 pm, about 0.1 pm to about 150.0 pm, about 0.2 pm to about 150.0 pm, about 0.2 pm to about 100.0 pm, about 0.2 pm to about 75.0 pm, or about 0.2 pm to about 50.0 pm. In some embodiments, a particle or microparticle can be about 0.01 pm to about 1,000 pm before, during and / or after coating is completed by an ALD system during the processes disclosed herein. In other embodiments, an encased or fully coated plurality of therapeutic agent-, antigen-, immunogen-, compound-containing or combination therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles having multiple layers is less than about 5.0 pm in size. In certain embodiments, the particles or microparticles, size prior to coating can be from about 1.0 pm to about 40.0 pm. It will be recognized that the elements of the therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles, coating layers, and any additional layers can be provided in concentrations capable of providing a suitable dose of a substance while maintaining an appropriate microparticle or particle size and maintaining syringability.

[0084] In certain embodiments and further to paragraphs

[0042] -

[0083] above, one advantage of using one or more aluminum-based materials as coating layer(s) is that the aluminum-based materials can also act as an adjuvant. In accordance with these embodiments, the aluminum-based coating layers surrounding the agent containing particles and microparticles expose essentially the same surface chemistries to immunoactive cells as do standard aluminum-based adjuvant particles known in the art. In certain embodiments, the aluminum-based coating layer can be sufficiently thin so that the total aluminum concentration per administration of the composition to a subject is less than about 100 pg. For example, the amount can be less than about 50 pg, less than about 40 pg, less than about 30 pg, less than about 20 pg, less than about 10 pg, less than about 5 pg, or less than about 1.0 pg or even less.

[0085] In some embodiments and further to paragraphs

[0043] -

[0084] above, coating layers other than aluminum-based coating layers can be used in order to coat the therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles. In accordance with these embodiments, non-aluminum coating layers including, but not limited to, silicon dioxide (SiCh), titanium dioxide (TiCh), zinc oxide (ZnO), or other metal oxide or other alkoxide can be used either in combination with aluminum-based coating layers, or alone to the exclusion of aluminum-based coating layers or as a combination of non-aluminum coating layers such as a mixed layer of silicon dioxide (SiCh), titanium dioxide (TiCh), and / or zinc oxide (ZnO). In some embodiments, a therapeutic agent-, antigen-, immunogen-, compound-containing microparticle can be coated with one or more aluminum-based layers, followed by one or more layers of a different material or they can be alternating. In certain embodiments, sequentially altering layers can be applied such as repeated application of for a predetermined number of AI2O3 layers, followed by the same or different number of TiCh layers (e.g., 5:5, 5:4, 10:10 or any other suitable alternating ratio) to reach the desired number of coating layers of a single or multiple layering material for coating a particle or microparticle disclosed herein. In accordance with these embodiments, different materials or repetitive layered structures formed from these materials can dissolve more slowly than the aluminum-based coating layer.

[0086] In other embodiments and further to paragraphs

[0043] -

[0085] above, one or more coating layers can be deposited on a therapeutic agent-, antigen-, immunogen-, compoundcontaining particle or microparticle by, for example, atomic layer deposition (ALD, for example any instrumentation capable of atomic layer deposition can be used). ALD includes a thin filmdeposition technique that is based on the sequential use of a gas phase chemical process. ALD is considered a type of chemical vapor deposition. In certain methods, a majority of ALD reactions use two chemicals, referred to as precursors. These precursors react with the surface of a material one at a time in a sequential, self-limiting, or directed manner. Through the repeated exposure to separate precursors, a thin film can be deposited. Use of an ALD system for example, ALD to deposit coating layers on antigen-, compound-, agent-containing particles or microparticles can be based on sequential, self-limiting reactions and provides for layer thickness control at the Angstrom level and tunable coating layer composition. Examples of ALD procedures of use in methods disclosed herein for depositing coating layers on agent-containing microparticles or particles can be found for example in LF Hakim et al, Adv Funct Mater, 2007 Nov; 17 (16):3175-81, DM King et al., Powder Technol, 2012 Can;22L 13-25, and X Liang etal., ACS Appl Mater Interfaces, 2009 Sept(web); 1(9): 1988-95, each of which is hereby incorporated by reference in their entirety.

[0087] Embodiments of the present disclosure and further to paragraphs

[0043] -

[0086] above, can include polypeptides, polynucleotides, carbohydrates, proteins, virus-like particles, inactivated or attenuated pathogens (e.g., live, attenuated viruses), or other antigens or agents that elicit a therapeutic response when introduced to a subject. In accordance with these embodiments, an elicited response can be a prophylactic response, reducing or preventing infection, disease, or toxicity induced by exposure to a pathogen including, but not limited to, a virus, bacteria, or fungus, or toxin, and / or can be therapeutic, reducing the severity, preventing, treating cancer, or treating an infection, disease, other health condition, or toxicity.

[0088] In certain embodiments and further to paragraphs

[0043] -

[0087] above, compositions, methods, and uses disclosed herein concern using a particle or microparticle coating system having a mechanically agitated reactor in combination with milling agents within the coating chamber that results in sheering, mixing, or impacting the particles or microparticles with milling agents for preventing, reducing, or eliminating agglomeration and / or adherence of nanoparticles, microparticles, particles, coated nanoparticles, coated microparticles or coated particles. In one embodiment, fluidization of a system contemplated herein uses a gas stream to agitate a powder bed containing particles or microparticles and milling agents for coating in the system in addition to the mechanical agitation by external devices such as vibrators and impactors or internal devices such as stirrers or impellers. The fluidized bed formed by the gas stream can be presentduring purge phases (e.g., purge gas such as argon only flowing through the system), and during application of precursors, which can be flowed through the system by the flow of the purge gas. As such, the systems and methods described herein utilize milling agents in the form of inorganic or organic, or spherical, cubical, or other geometric-shaped particles to assure adequate mixing, deagglomeration, and contact of the chemical reactants with all surfaces of individual particles for improved coating with reduced adherence and agglomeration.Compositions

[0089] Certain embodiments and further to paragraphs

[0043] -

[0088] above, relate to mixtures of a plurality of pharmaceutical agent-containing particles or microparticles and a plurality of milling agents. In accordance with these embodiments, mixtures can be essentially dry mixtures of powders or compositions. In certain embodiments, the essentially dry pharmaceutical agentcontaining particles or microparticles can be minimally agglomerated particles or microparticles, optionally less than 1.0% of the essentially dry pharmaceutical agent-containing particles or microparticles are agglomerated when in the presence of milling agents contemplated herein. In accordance with these embodiments, the essentially dry mixture can include a plurality of dry or essentially dry therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles and a plurality of milling agents to form, for example, a powder bed mixture. In some embodiments, these compositions can include a mixture of uncoated essentially dry therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles and / or minimally agglomerated coated essentially dry therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles and at least one milling agent or a plurality of milling agents. In some embodiments, coated essentially dry therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles can include a core, one or more coating layers, and a plurality of milling agents in a combination. It is contemplated herein that the plurality of milling agents can be removed from a mixture contemplated herein by methods disclosed herein in order to be stored, transported or used immediately in a pharmaceutical composition for administration to a subject.

[0090] In certain embodiments and further to paragraphs

[0042] -

[0089] above, milling agents of the plurality of milling agents alone or in a composition contemplated herein can include solid milling agents. In some embodiments, the solid milling agents can include one or more of inorganic or organic solid particles. In certain embodiments, the plurality of milling agents caninclude at least a first inorganic material, a second inorganic material, at least a first organic material, and / or a second organic material. In some embodiments, size of the first inorganic material and the second inorganic material are different. In some embodiments, size of the first organic material and the second organic material are different. In other embodiments, size of the first inorganic material and the second inorganic material can be the same. In some embodiments, size of the first organic material and the second organic material can be the same or different. In some embodiments, milling agents can include, but are not limited to, one or more of ZrCh, ZrCh / SiCh, Fe, AI2O3, iron oxide, glass, quartz, synthetic polymers, natural polymers, starch, cellulose, Teflon, polyethylene, Nylon, a large, spray-dried milling media particle or other similarly solid material or combinations thereof. In certain embodiments, the milling agent includes at least ZrCh / SiCh or similar agent thereof. In some embodiments, milling agents can include a three-dimensional shaped milling agent of materials or milling agents disclosed herein (e.g., ZrCh / SiCh). In accordance with these embodiments, three-dimensional shaped milling agents can include, but are not limited to, a sphere, a cube, a cone, a pyramid, a square, or other shape of uniform milling agents or a mixture of irregular shapes or a mixture of two or more shapes of the same or different sizes.

[0091] In certain embodiments and further to paragraphs

[0043] -

[0090] above, mass ratio of the one or more milling agents or plurality of milling agents to: uncoated, partially coated, or coated therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles can be about 1 : 10 up to about 50: 1. In certain embodiments, mass ratio can range from about 1:10, about 2:10, about 3:10, about 4:10, about 5:10, about 6:10, about 7:10, about 8:10, about 9:10, about 1:1, about2:l, about 3:l, about 4:1, about 5:l, about 6:l, about 7:l, about 8:1, about 9:1, about 10:1, about 11:1, about 12:1, about 13:1, about 14:1, about 15:1, about 16:1, about 17:1, about 18:1, about 19:1, about 20:1, about 21:1, about 22:1, about 23:1, about 24:1, about 25:1, about 26: 1, about 27: 1, about 28: 1, about 29: 1, about 30: 1, about 31 : 1, about 32: 1, about 33:1, about 34:1, about 35:1, about 36:1, about 37:1, about 38:1, about 39:1, about 40:1, about 41:1, about 42:1, about 43:1, about 44:1, about 45:1, about 46:1, about 47:1, about 48:1, about 49:1, or about 10:1. In some embodiments, the ratio can range from about 1:5, about 1:3, about 1:2, about 1:1, about 2: 1 or about 3:1, milling agents to therapeutic agent-, antigen-, immunogen-, compound-containing particles. In some embodiments, the ratio can range from about 1:1, about2:1 or about 3:1, milling agents to therapeutic agent-, antigen-, immunogen-, compoundcontaining particles.

[0092] In some embodiments and further to paragraphs

[0043] -

[0091] above, milling agents can be about 2 to about 20, about 2 to about 15, about 2 to about 10 times larger than an average size of the pharmaceutical agents containing particles or microparticles. In some embodiments, milling agents can be at least 2 times larger than fully coated, partially coated or therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles.

[0093] In some embodiments and further to paragraphs

[0043] -

[0092] above, essentially dry or dry or glassy therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles can include a core. In other embodiments, the core can include one or more of immunogenic agents, small molecule therapeutics, anti-cancer agents, anti-inflammatory agents, anti-immune agents, other therapeutics, and / or other biologies and combinations thereof. In some embodiments, immunogenic agents can include, but are not limited to, a pathogenic organism-derived antigen; for example, at least one of a viral antigen, bacterial antigen, toxin, fungal agent, recombinant peptide, recombinant protein, peptide derived from a target protein or pathogen, synthetic peptide or protein, a polynucleotide or fragment thereof (e.g. DNA, mRNA, RNA, siRNA, or chimera thereof), or combination thereof. In some embodiments, immunogenic agents can include, but are not limited to, at least one of a virus, a virus-like particle, a live virus, a live attenuated virus, an inactivated virus, a bacterial antigen, a bacteriophage, a phage, polypeptide or polynucleotide thereof, or a combination thereof. In some embodiments, pharmaceutical agents can include one or more polypeptides or fragments thereof, a polynucleotide, a whole organism, a small molecule, a chemical agent or a combination thereof.

[0094] In some embodiments and further to paragraphs

[0043] -

[0093] above, the therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles can be glassy particles or microparticles. In some embodiments, the therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles can be minimally agglomerated particles or microparticles, for example, when in the presence of one or more milling agents. For example, less than 10.0% or less than 5.0% or less than 1.0% of the therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles are agglomerated.

[0095] In some embodiments and further to paragraphs

[0043] -

[0094] above, the therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles can include atleast one coating layer or at least 10 coating layers or at least 50 coating layers or at least 100 coating layers or more, or any number in between or higher. For example, the coating layer can include at least one metal agent (e.g. metal oxide or metal alkoxide). In some embodiments, the oxide can include a metallo-organic material, metal oxide, non-metal oxide, metal alkoxide, silicon oxide, or a combination thereof. For example, the metal oxide or metal alkoxide can include, but is not limited to, one or more of aluminum oxide (AI2O3), an aluminum alkoxide, silicon dioxide (SiCh), Zinc dioxide (ZnCh), titanium dioxide (TiCh), and silicon nitride (SisN4) or combinations thereof, or mixtures thereof, or alternating layers thereof. In some embodiments, the metal alkoxide can be aluminum alkoxide. In some embodiments, the non-metal oxide can be silicon oxide. The silicon oxide can be silicon dioxide (SiCh).

[0096] In some embodiments and further to paragraphs

[0043] -

[0095] above, the present disclosure is directed to a mixture of minimally agglomerated coated therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles. In other embodiments, the coated therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles can include a core, one or more coating layers, and a plurality of milling agents.Methods

[0097] In certain embodiments, the present disclosure is further directed to methods for preparing minimally agglomerated uncoated, partially coated or fully coated pharmaceutical agent-containing particles or microparticles in a chemical vapor deposition chamber. In some embodiments, methods can include, but are not limited to, introducing a plurality of milling agents to a coating chamber of the chemical vapor deposition system, introducing a plurality of essentially dry or dry therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles to the coating chamber of the coating chamber, and coating the plurality of essentially dry or dry therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles with the metal oxide or metal alkoxide forming the minimally agglomerated coated essentially dry or dry therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles. In some embodiments, methods can further include mechanically agitating the plurality of milling agents within the reaction chamber of the coating chamber during coating of the plurality of essentially dry or dry therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles with a metal oxide or metal alkoxide. In other embodiments, minimally agglomerated coated therapeutic agent-, antigen-, immunogen-,compound-containing particles or microparticles and the plurality of milling agents can be removed from the coating chamber and separated from one another by methods disclosed herein. In some embodiments, the minimally agglomerated coated therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles can be re-introduced to the coating chamber for further coating in absence of the plurality of milling agents. In some embodiments, a different type of milling agent than initially introduced can be introduced for further coating with the minimally agglomerated coated therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles. In some embodiments, coating of the therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles can be conducted under vacuum or elevated pressures using chemical vapor deposition methods. In other embodiments, coating of the therapeutic agent-, antigen-, immunogen-, compoundcontaining particles or microparticles can include coating of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles using atomic layer deposition (ALD). In certain embodiments, the coating chamber can be a fluidized bed reactor, a gas-phase powder reactor, or a rotary reactor.

[0098] In some embodiments and further to paragraphs

[0043] -

[0097] above, the plurality of milling agents and the plurality of therapeutic agent-, antigen-, immunogen-, compoundcontaining particles or microparticles can be agitated by a flowing gas stream, mechanical vibration of the reactor, rotary motion of the reactor, magnetic field agitation in case of ferromagnetic particles, stirrers or impellers in the reactor bed, inducing sufficient mechanical movement to continuously break apart agglomerates of the plurality of essentially dry or dry therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles to be coated. In other embodiments, the plurality of milling agents and the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles can be removed from the reaction chamber of the gas-phase powder reactor after at least one coating layer is applied to the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles before depositing at least a final coating layer onto the plurality of essentially dry or dry therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles. In some embodiments, the milling agents can be separated from a partially coated plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles after being removed from the reaction chamber.

[0099] In some embodiments and further to paragraphs

[0043] -

[0098] above, either within the reaction chamber or outside of the reaction chamber, the plurality of milling agents and the plurality of coated or partially coated essentially dry or dry therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles can be separated based on physical properties. The physical properties include but are not limited to size, density, magnetic, ferromagnetic, or a combination thereof. In some embodiments, either within the reaction chamber or outside of the reaction chamber, the plurality of milling agents and the plurality of pharmaceutical agent-containing particles or microparticles can be separated by sieving, fluid dynamic methods, or using magnetic forces. In some embodiments, separation of the milling agents from a limited number of coating layers applied or all coating layers applied to a therapeutic agent-, antigen-, immunogen-, compound-containing particle or microparticle is by magnetic separation or by sieve or both.

[0100] In some embodiments and further to paragraphs

[0043] -

[0099] above, methods for preparing coated therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles can include, but are not limited to, introducing a plurality of milling agents on a mesh in a coating or reaction chamber of the chemical vapor deposition system, introducing a plurality of essentially dry or dry therapeutic agent-, antigen-, immunogen-, compoundcontaining particles or microparticles to the coating or reaction chamber of the chemical vapor deposition system, and coating the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles with the metal oxide or metal alkoxide forming minimally agglomerated coated essentially dry pharmaceutical agent-containing particles or microparticles. In certain embodiments, a mesh can be included to separate particles being coated from a milling agent contemplated herein. In accordance with these embodiments, the mesh can be metal wire mesh or a plastic mesh. In other embodiments, the mesh size can range from about 2 mesh to about 1000 mesh. For example, the mesh size can range from 5 mesh to 950 mesh, 10 mesh to 900 mesh, 15 mesh to 850 mesh, 20 mesh to 800 mesh, 25 mesh to 750 mesh, 30 mesh to 700 mesh, 35 mesh to 650 mesh, 40 mesh to 600 mesh, 45 mesh to 550 mesh, 50 mesh to 500 mesh, 55 mesh to 450 mesh, 60 mesh to 400 mesh, 65 mesh to 350 mesh, 70 mesh to 300 mesh, 75 mesh to 250 mesh, 80 mesh to 200 mesh, 85 mesh to 150 mesh, or 90 mesh to 100 mesh. In some embodiments, the mesh opening can range in size from about 1 micron to about 200 microns, about 10 microns to about 150 microns, about 20 microns to about100 microns, or about 30 microns to about 50 microns. Tn some embodiments, mesh size is 75 microns.

[0101] In some embodiments and further to paragraphs

[0043] -

[0100] above, position of the mesh in or outside of a coating chamber can vary. In some embodiments, the mesh can be placed closer to the inlet of the coating chamber or closer to the outlet of the coating chamber separating the plurality of milling agents from the pharmaceutical agent-containing particles or microparticles to be coated. In some embodiments, the plurality of milling agents can be removed from a system after at least 10 to at least 20 coating cycles; optionally coating can continue after removal of the plurality of milling agents. In accordance with these embodiments, particles or microparticles being coated can remain in a reaction chamber before and after milling agents are removed or particles or microparticles can be removed and reintroduced after they are separated from the milling agent and coating can resume.

[0102] As illustrated in FIG. 1, examples of an atomic layer deposition (ALD) system 100 is provided in a side cross-sectional view. The reactor vessel 104 extends between the inlet opening 108 and the outlet opening 112. The reactor vessel 104 can include an inlet section 128 defined by one or more sidewalls 130 and in fluid communication with the inlet opening 108. The reactor vessel 104 can also include an outlet section 132 defined by one or more sidewalls 134 and in fluid communication with the outlet opening 112. The one or more sidewalls 130, 134 can be a cylindrical tube. The reactor vessel 104 can further include a coating chamber 136 positioned between the inlet section 128 and the outlet section 132. The coating chamber 136 includes an inlet filter 140 adjacent the inlet section 128 of the reactor vessel 104 and an outlet filter 142 adjacent the outlet section 132 of the reactor vessel 104. The inlet and outlet filters 140, 142 can act as a microparticle or nanoparticle barrier, but not as a sieve. Process gases can flow through the filters 140, 142, but microparticles or nanoparticles are retained between the two filters 140, 142. In certain embodiments, the inlet and outlet filters 140, 142 (also referred to as a “frit”, “frit disk” or “frit disk filter”) can be, for example, sintered stainless steel filters that can be sized to retain the microparticles or nanoparticles between the inlet and outlet filters 140, 142. In certain embodiments, the inlet and outlet filters 140, 142 can be frit disk filters with about 2 micrometer pores or other suitable pore size to maintain the microparticles or nanoparticles within the reaction chamber for optimal retention and coating. The coating chamber 136 can be removably coupled to the inlet and outlet sections 128, 132 of the reactor vessel 104. In certain instances,the coating chamber 136 can be removably coupled to the inlet and outlet sections 128, 132 of the reactor vessel 104 via clamp fittings 156. Rubber or other suitable gaskets can be utilized to provide hermetic sealing between the inlet section 128, outlet section 132, and the coating chamber 136. The clamp fittings 156 can be made of a metal that will be in intimate contact with the reactor walls when tightened and thus can transfer vibrations due to vibration motors attached to the reactor mounting from the coating chamber 136 to the outlet section 132 that contains the outlet filter 142 causing the outlet filter 142 to vibrate along with the outlet section 132. In other embodiments, one or more ultrasonic transducer(s) 158 can be mounted or affixed to different parts of the reactor assembly to provide additional high-frequency vibration of the ALD system. In another embodiment, one or more ultrasonic transducer(s) can be mounted directly to an ALD system, for example using a screw or other component, for example, using a weld-on screw.

[0103] As seen in FIG. 1, the microparticles 144 to be coated or being coated are located within the coating chamber 136 of the reaction vessel 104. One or more process gases 146 are introduced to the inlet opening 108 of the reactor vessel 104, thereby fluidizing the microparticles within the coating chamber 136 (e.g., fluid bed). The one or more process gases 146 pass through the coating chamber 136 and exits the reactor vessel 104 at the outlet opening 112. As illustrated in FIG. 1, the ALD system 102 includes an agitator 106 (e.g., ultrasonic agitator) coupled to the coating chamber 136 of the reactor vessel 104. In the embodiment illustrated in FIG. 1, the agitator 106 includes a sonicator or ultrasonic transducer 158. The ultrasound transducer 158 includes an end cap 176, one or more piezoelectric plates 148, electrodes 150, and a radiation head 152. The components can be coupled together via a screw or other coupling component (not shown in FIG. 1). When an oscillating voltage is applied to the electrodes 150, the piezoelectric plates 148 vibrate and generate acoustic energy, such as ultrasound energy (>20kHz). Stated differently, the ultrasound transducer 158 converts electrical energy to acoustic energy, which can reach ultrasonic ranges, in the form of acoustic or sound waves. The piezoelectric plates 148 change size and shape when the voltage is applied. As an example, when AC voltage is applied, the piezoelectric plates 148 oscillate and produce ultrasound energy accordingly. The ultrasound energy is transmitted through the radiation head 152 to a wave propagating structure 154, such as a clamp collar, which is affixed to the coating chamber 136. In this configuration, the ultrasound energy from the ultrasound transducer 158 is transmitted through the clamp collar 154 to the coating chamber 136 to inhibit the agglomerationof microparticles on the inner surfaces of the chamber walls 138. In certain instances, the ultrasonic transducer 158 can be a 40 kHz 60 W transducer. For example, the ultrasonic transducer 158 can be a YaeCCC (used at for example, 60W 40KHz / 60W) Ultrasonic Cleaning Transducer Cleaner or similar or substitutable device known in the art. The control board for the ultrasonic transducer can be, for example, a Power Driver Board 110V AC or other suitable control board.

[0104] In some embodiments, the ALD system 102 can include more than one agitator 106 (e.g., ultrasonic agitator), for example two agitators 106 or more, coupled to the reactor vessel 104. In accordance with this embodiment, two agitators 106 can include a sonicator and / or ultrasonic transducer 158. Each ultrasonic transducer 158 can have the same or similar characteristics as the ultrasonic transducer 158. Ultrasound energy generated from each ultrasound transducer 158 can be transmitted through each respective collar 154 associated with the reactor vessel 104 (e g, coating chamber 136, outlet section 132). In some embodiments, a first ultrasonic transducer 158 can be coupled to the coating chamber 136 and a second ultrasonic transducer 158 can be coupled to the outlet section 132. In other embodiments, the two or more ultrasonic transducers 158 can be coupled to the reactor vessel 104 (e.g., the inlet section 128, coating chamber 136, outlet section 132) or in any other logical configuration and at various locations without departing from the scope of this disclosure. It is contemplated that 2 transducers are not a required feature but an alternative feature where one transducer or more than 2 transducers are also contemplated.

[0105] FIGS. 1B-1C illustrate a perspective view of a disassembled reactor system and assembled reactor system 104. In FIGS. 1B-1C, A is the outlet section 132, B is the inlet section 128, and C is the coating chamber 136.

[0106] The milling agent can also be suspended on a mesh above the powder bed. The mesh size should be sufficiently small to contain the milling particles (i.e. 200 mesh or 75 micron for 100 micron) but large enough to allow powder including agglomerates elutriated by the purge gas to pass. At the gas flowrates typical for the coating process, most particles and agglomerates sufficiently small to be elutriated will eventually leave the particle bed temporarily, pass through the vibrating milling agents, and settle back down into the particle bed. During the brief contact with the vibrating milling particles, agglomerates can be deagglomerated and the resulting primary particles will eventually settle back into the powder bed.

[0107] In some embodiments, the coating chamber 136 can be as illustrated in FIGS. 2A-2C and 3A-3B. As illustrated in FIGS. 2A-2C, in some embodiments, the coating chamber 136 can include a milling screen (e.g., a mesh), 204. In some embodiments, a milling screen 204 can span the one or more sidewalls 130 of the coating chamber. The power bed 210 including uncoated, partially coated, or coated pharmaceutical agent-containing particles or microparticles is positioned or deposited on an inner base of the coating chamber. The distance 206 between the milling screen 204 and the base of the coating chamber can be varied. For example, as illustrated in 200a the milling screen 204 can be in the middle of the coating chamber. Alternatively, as illustrated in an exemplary set up depicted in 200b, the milling screen 204 can be closer to the outlet section 132. In another embodiment, the milling screen 204 can be closer to the inlet section 128 and closer to a powder bed 210 as illustrated in representative illustration of 200c. The milling agents 202 can be placed on the milling screen 204. Size of the milling screen or mesh can vary. In certain embodiments, size of the milling screen depends on size of milling agents 202. In these examples, diameter of the milling agents is larger than size of openings of a milling screen or mesh 204. Particles 208 to be coated or during coating can elutriate (e.g., pass through a milling screen 204) and recirculate below and above the milling screen or mesh 204. A milling screen size should be sufficiently small to contain the milling agents separate from a powder bed and bottom part of a reaction chamber as depicted (e.g., 200 mesh or 75 micron pores, for 100 micron milling agent) but large enough to allow powder including agglomerates to travel through the milling screen by force of purge gas. At gas flowrates typical for a coating process, most particles and agglomerates sufficiently small to be elutriated will eventually leave a powder or particle bed temporarily, pass through vibrating or agitated milling agents, and settle back down into the powder or particle bed. During brief contact with the vibrating milling agents, particle agglomerates can be deagglomerated and deagglomerated particles can settle back into the powder or particle bed.

[0108] As illustrated in FIG. 3A, in some embodiments, the milling screen 204 can be separated from the coating chamber 136. Reactor systems disclosed herein and of use for scale-up can have more than one unit stacked upon one another to create a reaction system having a coating chamber as illustrated herein. In some embodiments, the reactor system can have four units as depicted in FIG. 3B. The four units can include a top unit 132, a coating chamber 136 having 2 combined units (136), and a bottom unit 128. A milling screen 204 can be attached to the bottomof a top unit of reaction chamber 136 or the top of a bottom unit of reaction chamber 136.Attachment can occur by any means such as by removable method or by glue or other suitable adhesive or can be permanently affixed by appropriate means (e.g., brazing for a metal milling screen). In some embodiments, the milling screen can be removably fixed in position using one or more flanges and clamps alone or in addition to removably adhering the milling screen 204 to a surface of a unit as indicated herein. Rubber or other suitable gaskets can be utilized to provide sealing (e.g., hermetic or other) of the milling screen 204 between the units of the coating chamber 136, to reduce or eliminate gases, milling agents and / or particles from escaping from the reaction system. The units can be assembled with any known flange. In some embodiments, a 2-micron sintered metal frit / filter can be removably attached, glued, adhered or permanently affixed to the bottom unit to keep a powder bed in the coating chamber 136. In certain embodiments, a sintered metal frit / filter (e.g., 2 micron) can also be attached, glued or welded to the top unit to keep the particles or microparticles to be coated within the coating chamber 136. Flanges can be assembled with clamps and rubber gaskets as depicted herein. As illustrated in an exemplary set up depicted in 300b, a milling screen 204 can be absent from the coating chamber 136 and milling agents 202 and uncoated, partially coated, or coated pharmaceutical agent-containing particles or microparticles 208 intermixed. In some embodiments, milling agents 204 are heavier or denser than an average uncoated, partially coated, or coated pharmaceutical agent-containing particles or microparticles. In other embodiments, milling agents 204 are similar density or a lower density than an average uncoated, partially coated, or coated pharmaceutical agent-containing particles or microparticles.

[0109] Other embodiments and further to paragraphs

[0042] -

[0108] above, provide kits of use with the disclosed mixtures and / or kits for coating pharmaceutical agent-containing particles. In certain embodiments, a kit can include milling agents, for example, inorganic or organic spherical particles for inclusion in the coating chamber of the ALD system. In other embodiments, kits can further include a sieve or magnet to separate milling agents from the coated particles or microparticles at the final round or prior to the final rounds of coating of the particles or microparticles as disclosed herein. In other embodiments, kits can be portable for storage and transport of systems and components disclosed herein. In some embodiments, kits can include an ALD system, a plurality of milling agents, a plurality of pharmaceutical agentcontaining particles and microparticles, and instructions for using the ALD system.EXAMPLES

[0110] The materials, methods, and embodiments described herein are further defined in the following Examples. Certain embodiments are defined in the Examples herein. These Examples, while indicating certain embodiments, are given by way of illustration only. From the disclosure herein and these Examples, one skilled in the art can ascertain the essential characteristics of this invention, and without departing from the spirit and scope thereof, can make various changes and modifications of the invention to adapt it to various usages and conditions.[OHl] In one exemplary method, testing was performed on experimental powders using examples of the systems and methods described herein. First, experimental powders were obtained, as discussed below in the Example - Powder Sample Preparation section. In one set of examples, individual powder samples were used in various coating processes (e.g., number of ALD cycles, with milling agents added to the coating chamber. In a first example, ALD coating for 100 cycles was performed with milling agents within the coating chamber (as discussed below in Example 1).

[0112] In another example, coating quality of the coated powder samples were assessed with dissolution assays in aqueous buffer solutions. The properties of the powder samples as a function of the coating process are illustrated in the table in FIG. 1 below. In some examples, ALD coating was performed for 50 alumina ALD cycles without milling agents. In certain examples, ALD coating was performed for 50 alumina ALD cycles with milling agents, followed by 5- alumina ALD cycles without the milling agent. In another example, photographic images were collected to illustrate the degree of agglomeration, and the particle size distribution showed the degree of agglomerates in the final product after coating the particles with milling agents present in the coating chamber. Additional testing was performed (as discussed below in Examples 2 and 3).Exemplary - Powder Sample Preparation

[0113] Experimental powders were obtained. In these example methods, powders with a composition of approximately 70-90 wt% trehalose, 10-25 wt% hydroxyethyl starch, and 5-10 wt% other minor components (0-5% histidine, 0-5% ammonium acetate, 0-1% APIs) were prepared by spray drying (e.g., with a Buechi B-290 Mini Spray Dryer (Buchi Labortechnik AG, Flawil, Switzerland) fitted with a two-fluid nozzle at an inlet temperature of 60-100 °C for the drying gas, an outlet temperature of 40-60° C, and an atomizer flow rate of 5-10 liters / minute forthe spray nozzle). The powders were further dried in a lyophilizer FTS Systems Lyophilizer at 60 mTorr for 16 h at 40° C to a residual moisture content of less than 1% as determined by Karl-Fischer titration.Example 1Coating spray dried powders with AI2O3 in a fluidized bed using ZrCh / SiOi beads

[0114] In one exemplary method, 2 g of a spray-dried powder formulation with a mean particle size of 10 micron and a density of 1.5 g / cm3was thoroughly mixed with approximately 6.0 g of 0.1 mm spherical ZrCh / SiCh or beads (density of ~3.7 g / cm3) to obtain a homogeneous phase. Homogenizing was carried out in a vial with a vortexer, by gentle mixing with a mortar and pestle, or any other suitable homogenization method to break up any large clumps of the starting material without damaging the particles. The homogenized mixture was added to the fluidized bed ALD reactor and after appropriate drying times to remove residual solvent from the spray drying process under vacuum without agitation or gas flow (typically between 1 and 3 hours), and slow heating to the reaction temperature, the ALD coatings were carried out.

[0115] Spray-dried powders were coated with alumina (AI2O3) in a custom-built ALD fluidized bed coating chamber. Briefly, spray-dried particles were exposed to alternating pulses of TMA (Sigma Aldrich) and water under argon at 2 to 3 torr and 50 °C. Each TMA-water cycle deposits a single-molecular layer of alumina with an expected thickness of 2.3 A. To reduce agglomeration during coating, 0.1 mm zirconia / silicon beads were mixed with the spray-dried powders at a 3 : 1 weight ratio prior to ALD coating. Beads were removed after the first 20 cycles by sieving. A total of 30 or 50 cycles ALD cycles were applied to the powder.

[0116] The AI2O3 layers were formed according to the net reaction 2 A1(CH3)3 + 3H2O -> AI2O3 + 6 CH4. That reaction was split into two half reactions for the ALD process with sequential exposure of the powder surface to Al(CFb)3 (trimethylaluminum, TMA) and H2O vapors, where a single layer of AI2O3 with a thickness between 0.15 and 0.3 nm was deposited after each water and TMA sequence. The number of TMA / water cycles applied then determined the final coating thickness. For example, 100 coating cycles deposited AI2O3 with a thickness of about 20-30 nm. For the examples described herein, the powders were exposed to the ALD precursors in a 0.875” I.D. 2” long tubular stainless-steel reactor with 2-micron sintered metal frit filter disks at the reactor inlet and outlet. Trimethylaluminum vapor was fed at a flowrate of ~1 cm3(STP) for 1 min, followed by 4 min of purge with dry Argon. Then ~1 cm3(STP) of water vapor was added ata flowrate of ~1 cm3(STP) for 1.3 min to maintain the stoichiometric ratio of the reaction. The reactor was then purged with dry Argon for 6 minutes before the cycle was repeated. A constant Argon flow rate of 1.5 cm3(STP) was maintained throughout the deposition process, both for the purge steps and during dosing. The background pressure during the Argon purge was in the range of 2-4 torr and the pressure increases by ~1.5 torr during the precursor dosing. Continuous vibration and intermittent mechanical impact and ultrasonic vibration during the coating process assured continuous deagglomeration by the milling agents, minimized wall adhesion, and even coating of primary particles.

[0117] After a sufficient number of ALD cycles were deposited (e.g., about 10, about 20, about 30), the coating process was interrupted, and the milling agents were removed by sieving. In these examples, if desired, the coating process of the particles can continue for more cycles without milling agent being present.Example 2Evaluation of coating quality using dissolution assays

[0118] In another exemplary method, coating quality of particles was assessed with dissolution assays in aqueous buffer solutions. AI2O3 slowly converts to loosely crystallized AIPO4 in slightly acidic phosphate buffers (pH of 6.5). After this conversion has penetrated the full thickness of the AI2O3 layer, water can permeate to the particle core resulting in rupture of the alumina shell and release of the content into the buffer solutions. The time dependance of this release can be measured if appropriate tracers are present in the particle core. The release of 1-histidine, a typical buffer for spray drying of pharmaceutical powders, can easily be detected with a colorimetric OPA assay (OPA is o-phtalaldehyde). Well coated powders with few agglomerates do not dissolve for several hours, depending on the concentrations and temperatures of the buffer, and complete dissolution is eventually achieved in a relatively step over a short time interval. The buffer properties (pH, concentration, chemical composition, temperature) can be optimized to result in dissolution within a reasonable timeframe for a feasible routine assay, for example within one day for 20 nm coatings. FIG. 4 compares a time dependent dissolution of spray-dried powders containing some 1-histidine as tracer that were coated with 50 AI2O3 ALD cycles with and without 0.1 mm ZrCh milling agents present for some of the coating cycles. The powder coated without the milling agents had about 5% of is content released immediately, followed by an initially slowly increasing rate of release with timeand a steeper step with a half-life of about 16 hours. The milled powder of this example, where milling agents were only present in the powder bed during an initial 20 AI2O3 ALD cycles, in contrast, did not show any measurable dissolution until about 12 hours, half of the powder content dissolved after about 22 hours and nearly complete dissolution was achieved at about 24 hours. The alumina weight loading for both powders was similar at ~7 wt%, and thus, the milled powder was about 30% more protective at the same coating thickness. The more uniform and prolonged protection of the milled powders is likely due to a more uniform coating thickness as a result of fewer dynamically forming and degrading agglomerates during the coating cycles. Example 3Evaluating presence of agglomerates after milling using a particle size analyzer

[0119] In yet another method, presence of agglomerates in the product was determined with liquid-phase particle size analyzers such as FlowCam (Yokogawa Fluid Imaging Technology). Dry powder light scattering methods provide primary particle size distributions but the aggressive aerosolization process used by most instruments is sufficiently abrasive to deagglomerate the coated clusters and thus the results don’t reflect the relevant size distribution that indicates the presence of agglomerates sufficiently stable to affect product performance in liquid environments. FIGS. 7A-7B compare example images measured by FlowCam for powders coated with and without 0.1 mm ZrCh / SiCh milling agents present during the coatings. Whereas the milled powders are composed mostly of spherical primary particles, the powders coated without milling agents contain a large fraction of agglomerates that are sufficiently stable to maintain their integrity in the liquid, even when ultrasonic vibration is applied for better dispersion prior to the measurements.

[0120] The FlowCam software calculates size distributions from the images as shown in FIGS.7A-7B. The size distribution of the spray dried powder prior to the ALD coatings measured by the FlowCam is similar to that determined with a dry dispersion light scattering instrument (Mastersizer 2000, Malvern Panalytical). The FlowCam size limit is about 1 mm and thus the sub-micron fines detected by the Mastersizer are not shown in the FlowCam distribution. The FlowCam size distribution of a powder coated without the milling agents shown in FIG. 5A is shifted to larger particle sizes by about 5 mm as expected from the images in FIG. 5B. Similarly, the SEM images of a spray-dried powder coated without milling agents (FIG. 7A) show several larger particle clusters whereas the same powder coated in the presence of 100 mm ZrCh / SiChspheres is mostly composed of individual particles. The particle size distribution of the spray-dried powders was also measured with a Malvern Analytical Mastersizer 2000 laser particle size analyzer (Malvern, UK) before and after coatings. The size distributions measured after coatings (not shown), were similar to those of the starting materials in all cases, with and without milling agents since the aggressive aerosolization process of the instrument deagglomerated any particle clusters prior to detection. The particle sizes of the spray dried sample have bimodal distribution with a small fraction of sub-micron particles and an average size of a majority of particles near 8-10 pm. The BET surface area of approximately 1 m2 / g was consistent with the size distribution. The majority of sub-micron fines, however, disappeared in the Mastersizer distribution of the coated powders. The loss in sub-micron fines is mostly due to adhesion of those fines into depressions in the irregular surface of the spray dried particles as shown in the SEM images in FIGS. 7A-7B and these depressions are not even accessible by the much larger milling agents. Powders coated with the milling agents show FlowCam distributions much closer to those of the starting materials, indicating that the milling agents minimize the formation of larger agglomerates. The type of milling agents affects these size distribution and larger and heavier particles such as the 0.1 mm ZrCh / SiCh spheres with a density of 3.7 g / cm3show a larger shift in sizes after 20 of 100 cycles than 0.05 mm glass beads with a density of ~2.5 g / cm3, even though the glass beads still improve the size distribution over coatings carried out without milling.Example 4Coating spray dried powders with AI2O3 in a fluidized bed with a milling screen using ZrCh / SiOi Beads

[0121] In one exemplary method, 1 g of spray-dried powder formulation and about 3 g of 0.1 mm spherical ZrCh / SiCh beads were added to a 6-inch long 1-inch ID reactor with a 200-mesh sieve mounted at a height of 2 inches. Initially the powder / milling agent mix resided on the mesh. As in example 1, the mixture was dried under vacuum to remove residual solvent from the spray drying process and then 50 AI2O3 ALD layers were deposited. The milling agent either remained present throughout all 50 coating cycles or they were removed after 20 cycles and the coating was continued for 30 more coating cycles. Only a small fraction of powder (<1%) was present on the milling agent bed after they were removed following 20 or 50 ALD cycles because the majority of particles quickly sieved through the large mesh after the coating wasinitiated and then remained at the bottom of the coating chamber. A small portion of the temporarily elutriated and circulating particles were found clinging to the milling agents.

[0122] As illustrated, dissolution curves represented in FIG. 8 illustrate that a fraction of the coated particles can be damaged but resolve if milling beads are present throughout (e.g., during at least 10 cycles, and as illustrated here, during some or all 50 cycles) indicating that the elutriation and milling occurs continuously to deagglomerate agglomerates, affecting all particles, but also causes some damage to the coating layer. When the milling agents were present for only 20 coating cycles, sufficient deagglomeration and uniform AI2O3 coating occurs to minimize further agglomeration during the 30 consecutive cycles to complete the coating of alumina layer, similar to the examples where the milling agents are mixed with the powder. Only a small fraction (<5%) dissolve during the first few hours for that sample. In this example, one test used milling agents present for 20 ALD coating cycles and then 30 cycles without milling media; 20 ALD coating cycles with the milling media present suspended on mesh and 30 cycles without milling media; or 50 coating cycles with the milling media present suspended on mesh for the entire 50 cycles and then these parameter and results were assessed, see for example FIG.8.

[0123] In this example, the fraction of damaged particles dissolving initially was even lower (<2%) when the first twenty coatings were carried out with the milling agents and powder mixed, similar to example 1, but the overall time dependance of the dissolution behavior was close to that of the powder coated with the milling agents suspended on the mesh for 20 cycles. In contrast, about 10% of particles dissolved at the same time for the sample that was coated for 50 cycles without milling agents present and 50% of the powder had dissolved ~ 4-5 hours before half of the particles milled for 20 cycles released their content in the buffer.All the compositions and methods disclosed and claimed herein can be made and executed without undue experimentation in light of the present disclosure. While the compositions and methods have been described in terms of embodiments, it is apparent to those of skill in the art that variations can be applied to the compositions and methods and in the steps or in the sequence of steps of the methods described herein without departing from the concept, spirit and scope herein. More specifically, certain agents that are both chemically and physiologicallyrelated can be substituted for the agents described herein while the same or similar results would be achieved. All such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the spirit, scope and concept as defined by the appended claims.

Claims

What is claimed is:

1. A method for making minimally agglomerated coated plurality of pharmaceutical agentcontaining particles or microparticles in a chemical vapor deposition system, the method comprising:introducing a plurality of milling agents into a coating chamber of the chemical vapor deposition system;introducing a plurality of therapeutic agent-, antigen-, immunogen-, compoundcontaining particles or microparticles to the coating chamber of the chemical vapor deposition system; andcoating the plurality of therapeutic agent-, antigen-, immunogen-, compoundcontaining particles or microparticles with at least one of a metal agent or non-metal agent and forming a minimally agglomerated coated plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles.

2. The method according to claim 1, further comprising mechanically agitating the plurality of milling agents and the plurality of therapeutic agent-, antigen-, immunogen-, compoundcontaining particles or microparticles within the coating chamber of the chemical vapor deposition system during coating of the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles.

3. The method according to claim 1 or 2, wherein the coating of the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles is conducted under vacuum or elevated pressures using chemical vapor deposition methods.

4. The method according to any one of claims 1 to 3, wherein the minimally agglomerated coated plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles and plurality of milling agents are removed from the coating chamber after at least 10 coating cycles; optionally, wherein the minimally agglomerated coated plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles are re-introduced into the coating chamber for additional coatings.

5. The method according to any one of claims 1 to 4, wherein the coating of the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles comprises coating of the plurality of therapeutic agent-, antigen-, immunogen-, compoundcontaining particles or microparticles using atomic layer deposition (ALD).

6. The method according to any one of claims 1 to 5, wherein the coating chamber comprises a fluidized bed reactor, a gas-phase powder reactor, or a rotary reactor.

7. The method according to any one of claims 1 to 6, wherein the plurality of milling agents and the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles are agitated by at least one of: a flowing gas stream, mechanical vibration of the coating chamber, rotary motion of the coating chamber, magnetic field agitation in case of ferromagnetic particles, stirrers or impellers in the coating chamber, inducing sufficient mechanical movement to intermittently or continuously break apart agglomerates of uncoated or partially coated particles of the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles.

8. The method according to any one of claims 1 to 7, wherein the plurality of milling agents and the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles are removed from the coating chamber of the chemical vapor deposition system after at least ten (10) coating layers are applied to the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles, and the plurality of milling agents are separated from a plurality of coated therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles; optionally, before depositing at least a final coating layer onto the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles using the chemical vapor deposition system.

9. The method according to any one of claims 1 to 8, wherein either within the coating chamber or outside of the coating chamber, the plurality of milling agents and a plurality of coated therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles are separated based on physical properties of the plurality of milling agentsand a plurality of coated therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles.

10. The method according to claim 9, wherein the physical properties comprise at least one of size, density, magnetic, ferromagnetic, or a combination thereof.

11. The method according to any one of claims 1 to 10, wherein either within the coating chamber or outside of the coating chamber, the plurality of milling agents and a plurality of coated therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles are separated by sieving, fluid dynamic methods, or using magnetic forces from the plurality of milling agents.

12. The method according to claim 9 or 11, wherein a separated coated minimally agglomerated plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles can further be stored for later use or form part of an essentially dry or rehydrated pharmaceutical composition, optionally, wherein the separated coated minimally agglomerated plurality of therapeutic agent-, antigen-, immunogen-, compoundcontaining particles or microparticles are re-introduced into the coating chamber for additional coatings.

13. The method according to any one of claims 1 to 12, wherein introducing the plurality of milling agents; and introducing the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles, to the coating chamber of the chemical vapor deposition system comprises at least one of a) premixing the plurality of the milling agents and the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles prior to introducing them to the coating chamber of the chemical vapor deposition system; b) simultaneously introducing the plurality of the milling agents and the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles; and c) separately introducing the plurality of the milling agents and the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles.

14. The method according to any one of claims 1 to 13, wherein milling agents of the plurality of milling agents comprise one or more of inorganic or organic milling agents; optionally, wherein the one or more of inorganic or organic milling agents comprise one or more of zirconium dioxide (ZrCh), zirconium dioxide / silicon dioxide (ZrCh / SiCh), iron (Fe), or aluminum oxide (AI2O3) glass, quartz, synthetic polymers, natural polymers, starch, cellulose, Teflon, polyethylene, Nylon other similar solid material or combinations thereof.

15. The method according to any one of claims 1 to 14, wherein the metal agent or non-metal agent comprises at least one of a metallo-organic material, a metal oxide, metal alkoxide, a non-metal oxide, a silicon oxide, or a combination thereof.

16. The method according to claim 15, wherein the metal oxide or the metal alkoxide comprises at least one of an aluminum oxide (AI2O3), an aluminum alkoxide, silicon dioxide (SiCh), Zinc dioxide (ZnCh), titanium dioxide (TiCh), and silicon nitride (Si3N4), and any combinations thereof.

17. The method according to claim 15 or 16, wherein the metal alkoxide comprises aluminum alkoxide and / or wherein the non-metal oxide comprises silicon dioxide (SiCh).

18. The method according to any one of claims 1-17, wherein milling agents of the plurality of milling agents comprise at least one of zirconium dioxide (ZrCh) and zirconium dioxide / silicon dioxide (ZrCh / SiCh).

19. The method according to claim 8, wherein the separated milling agents are replaced with at least one of smaller and less dense milling agents when the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles are reintroduced to the coating chamber before depositing at least one additional coating layer onto the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles.

20. A method for coating therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles in a chemical vapor deposition system, the method comprising:introducing a plurality of therapeutic agent-, antigen-, immunogen-, compoundcontaining particles or microparticles or combination thereof to a fluidized bed of a coating chamber of the chemical vapor deposition system;introducing a milling screen above the fluidized bed containing the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles or combination thereof;introducing a plurality of milling agents on the milling screen; andcoating the plurality of therapeutic agent-, antigen-, immunogen-, compoundcontaining particles or microparticles or combination thereof with at least one of a metal agent or non-metal agent using the chemical vapor deposition system.

21. The method according to claim 20, further comprising reducing or eliminating agglomeration of the plurality of therapeutic agent-, antigen-, immunogen-, compoundcontaining particles or microparticles or combination thereof during a coating process.

22. The method according to claim 20, wherein the milling screen comprises a mesh or separation feature to separate the plurality of milling agents from the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles while permitting intermittent contact of the plurality of milling agents with the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles during a coating process.

23. The method according to claim 22, wherein the mesh comprises at least one of a metal mesh, wire mesh or screen, or a plastic mesh or screen.

24. The method according to any one of claims 22-23, wherein the mesh further comprises pores and size of the pores of the mesh comprises from about 1.0 micron to about 200.0 microns.

25. The method according to claim 24, wherein a mesh pore size comprises about 20.0 to about 200.0 microns; optionally, wherein the mesh pore size is about 75.0 microns.

26. The method according to claim 24 or 25, wherein the size of the pores is sufficiently large to allow elutriating and recirculating the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles and microparticles or combinations thereof.

27. The method according to any one of claims 20 to 26, further comprising mechanically agitating the coating chamber during coating of the plurality therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles or combinations thereof.

28. The method according to any one of claims 20 to 27, wherein milling agents of the plurality of milling agents comprise a density higher than the density of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles or combinations thereof of the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles or combinations thereof.

29. The method according to any one of claims 20 to 27, wherein milling agents of the plurality of milling agents comprise a density lower than an average density of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles or combinations thereof of the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles or combinations thereof.

30. The method according to claim 28, wherein the higher density milling agents of the plurality of milling agents are present for a number of coating cycles and then removed and replaced with a plurality of milling agents having a lower density than an average density of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles or combinations thereof of the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles or combinations thereof; and further coating the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles in presence of the lower density milling agents.

31. The method according to any one of claims 20 to 30, wherein a diameter of the milling agents of the plurality of milling agents is larger than pores of the milling screen.

32. The method according to any one of claims 20 to 31, wherein a coating of the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles ormicroparticles is conducted under vacuum or elevated pressures using chemical vapor deposition methods.

33. The method according to any one of claims 20 to 31, wherein a coating of the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles or combination thereof comprises coating of the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles using atomic layer deposition (ALD).

34. The method according to claim 33, wherein number of ALD coating cycles ranges from about 10 cycles to about 2,000 cycles or more; or about 20 cycles to about 2000 cycles or more, or about 100 cycles to about 500 cycles.

35. The method according to claim 34 wherein the plurality of milling agents is at least one of removed from the chemical vapor deposition system and separated from the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles or combination after at least 10 cycles or at least 20 cycles.

36. The method according to claim 35, further comprising continuing coating the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles or combination thereof after removal of the plurality of milling agents.

37. The method according to claims 20 to 36, wherein the plurality milling agents present on the milling screen are removed from the reactor system after at least 10 cycles of coating, and at least one of smaller and less dense milling agents are introduced directly into a plurality of coated therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles or combination thereof to reduce or prevent agglomeration or to enhance deagglomeration during additional coating cycles to be performed.

38. The method according to any one of claims 20 to 37, wherein the coating chamber comprises a fluidized bed reactor.

39. The method according to any one of claims 20 to 38, wherein the plurality of milling agents and the plurality of therapeutic agent-, antigen-, immunogen-, compound-containingparticles or microparticles or combination thereof are agitated by at least one of a flowing gas stream, mechanical vibration of the coating chamber, rotary motion of the coating chamber, magnetic field agitation in case of ferromagnetic particles, stirrers or impellers in the coating chamber, and inducing sufficient mechanical movement to continuously break apart agglomerates of the plurality of therapeutic agent-, antigen-, immunogen-, compoundcontaining particles or microparticles or combination thereof to be coated.

40. The method according to any one of claims 20 to 39, wherein a fully coated plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles or combination thereof can be removed from the coating chamber and further be stored for later use or form part of a pharmaceutical composition.

41. An ALD system for coating a plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles or combination thereof, the ALD system comprising:an ALD system comprising a coating chamber configured to house the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles or combination thereof and a plurality of milling agents therein and the coating chamber configured to permit a flow of process gas therethrough to coat the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles or combination thereof;a gas inlet connected to the coating chamber;a gas outlet connected to the coating chamber; andone or more agitator(s) associated with or coupled to the coating chamber and configured to deliver mechanical energy to the coating chamber;wherein the one or more agitator(s) comprise one or more of a sonicator, and a wave propagating structure coupled to the sonicator and the coating chamber, one or more vibration motors, and a mechanical impactor configured to deliver mechanical energy; wherein the coating chamber is positioned between the gas inlet and the gas outlet of the ALD system; andwherein the coating chamber is optionally configured with a milling screen onto which the plurality of milling agents is placed.

42. The ALD system according to claim 41, wherein the ALD system comprises a milling screen and the milling screen comprises a mesh or plastic barrier.

43. The ALD system according to claim 42, wherein milling screen is positioned within the coating chamber sufficiently high above the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles or combination thereof to be coated to allow for particle bed expansion during a coating process.

44. The ALD system according to any one of claims 41 to 43, wherein a mesh contains pores and size of the pores is sufficiently small to suspend the plurality of milling agents above the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles or combination, and the size of the pores is sufficiently large to allow elutriating and circulation of the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles or combination thereof through the milling screen.

45. A kit comprising: a plurality of milling agents, at least one container, and instructions for using the plurality of milling agents in a chemical deposition system.

46. The kit according to claim 45, further comprising a plurality of essentially dry uncoated therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles or combination thereof.

47. A device comprising:chamber walls forming a coating chamber operable to house a plurality of pharmaceutical agent-containing particles or microparticles;an inlet at a first end of the coating chamber;an outlet at a second end of the coating chamber; anda milling screen disposed between the inlet and the outlet within the coating chamber to maintain separation of a plurality of milling agents from the plurality of pharmaceutical agent-containing particles or microparticles;wherein the coating chamber is configured to permit a flow of process gas to pass therethrough from the inlet to the outlet to coat the plurality of pharmaceutical agent-containing particles or microparticles to undergo a coating process in a fluidized bed environment.

48. The device according to claim 47, wherein the milling screen comprises a mesh or separation feature.

49. The device according to claim 48, wherein the mesh or separation feature comprises at least one of a wire mesh or screen or a plastic mesh or screen.

50. A mixture comprising a plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles or combination thereof and a plurality of milling agents.

51. The mixture according to claim 50, wherein the mixture is essentially dry.

52. The mixture according to claim 50 or 51, wherein a milling agents of the plurality of milling agents comprises solid agents.

53. The mixture according to any one of claims 50 to 52, wherein the milling agents of the plurality of milling agents comprise one or more of inorganic or organic milling agents.

54. The mixture according to any one of claims 50 to 53, wherein the milling agents comprise one or more of zirconium dioxide (ZrCh), zirconium dioxide / silicon dioxide (ZrCh / SiCh), iron (Fe), or aluminum oxide (AI2O3), iron oxide, glass, quartz, synthetic polymers, natural polymers, starch, cellulose, Teflon, polyethylene, Nylon, a large, spray-dried milling media particle, or other similarly solid material or combinations thereof; optionally, wherein the milling agents comprise one or more of zirconium dioxide (ZrCh), zirconium dioxide / silicon dioxide (ZrCh / SiCh), iron (Fe), or aluminum oxide (AI2O3), or iron oxide or a combination thereof.

55. The mixture according to any one of claims 50 to 54, wherein the milling agents comprise a three-dimensional shape; optionally, comprising a sphere, a cube, a cone, a pyramid, a square, or other shape of uniform milling agents or a mixture of shapes or mixture of irregular shapes or a mixture of sizes.

56. The mixture according to any one of claims 50 to 55, wherein a mass ratio of the plurality of milling agents to the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles or combination thereof ranges from about 1:10 up to about 50:1, optionally from about 1:1 to about 3:1 milling agent to particles or microparticles.

57. The mixture according to any one of claims 50 to 56, wherein the milling agents are about 2 to about 10 times larger than an average size of a therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles or combination thereof of the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles or combination thereof.

58. The mixture according to any one of claims 50 to 57, wherein the plurality of milling agents comprises one or more inorganic materials or one or more organic material.

59. The mixture according to any one of claims 50 to 58, wherein each of the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles or combination thereof are encased or stabilized in a matrix; optionally, a glassy matrix.

60. The mixture according to any one of claims 50 to 59, wherein plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles or combination thereof are minimally agglomerated particles or microparticles, optionally less than 1.0% of the plurality of therapeutic agent-, antigen-, immunogen-, compoundcontaining particles or microparticles or combination thereof are agglomerated compared to a mixture not including the plurality of milling agents.

61. The mixture according to any one of claims 50 to 60, wherein the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles or combination thereof are embedded in an organic glassy matrix comprising at least one glass-forming agent or shell-forming agent.

62. The mixture according to claim 61, wherein the at least one glass-forming agent or shell-forming agent comprises one or more of trehalose, sucrose, trileucine or other shortpeptide of 10 or less amino acids, ficoll, dextran, maltotriose, lactose, hydroxyethyl starch, glycine, glycine and mannitol, cyclodextrin, povidone, or a combination thereof.

63. The mixture according to any one of claims 50 to 62, wherein plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles or combination thereof comprise spray-dried, lyophilized, vacuum-dried, spray-freeze-dried or other evaporatively dried plurality of glassy plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles or combination thereof.

64. The mixture according to any one of claims 50 to 63, the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles comprises at least one coating layer applied by chemical vapor deposition; optionally, wherein the chemical vapor deposition comprises ALD.

65. The mixture according to claim 64, wherein the at least one coating layer comprises at least one of a metal agent, non-metal agent, or a combination thereof.

66. The mixture according to claim 65, wherein the metal agent comprises a metal oxide or a metal alkoxide.

67. The mixture according to claim 65, wherein the metal agent comprises at least one of aluminum oxide (AI2O3), an aluminum alkoxide, silicon dioxide (SiCh), Zinc dioxide (ZnCh), titanium dioxide (TiCh), and silicon nitride (SisN4), and any combinations thereof.

68. The mixture according to claim 66, wherein the metal alkoxide comprises an aluminum alkoxide.

69. The mixture according to claim 65, wherein the non-metal agent comprises silicon dioxide.

70. The mixture according to any one of claims 50 to 69, wherein a therapeutic agent, antigen, immunogen, and / or compound of the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles or combination thereof comprise at least one of immunogenic agents, small molecule therapeutics, anti-canceragents, anti-inflammatory agents, anti-immune agents, antibodies or fragments thereof, other therapeutics, and / or other biologies and combinations thereof.

71. The mixture according to claim 70, wherein the immunogenic agents comprise at least one of a viral antigen, bacterial antigen, toxin, fungal agent, recombinant peptide, recombinant protein, peptide derived from a target protein or pathogen, synthetic peptide or protein, a polynucleotide or fragment thereof, or combination thereof.

72. The mixture according to claim 70, wherein the immunogen comprises at least one of a virus, a virus-like particle, a live virus, a live attenuated virus, an inactivated virus, a bacterial antigen, a bacteriophage, a phage, a prion, or a combination thereof.

73. The mixture according to any one of claims 50 to 72, wherein a therapeutic agent, antigen, immunogen, and / or compound of the plurality of therapeutic agent-, antigen-, immunogen-, compound-containing particles or microparticles or combination thereof comprises at least one polynucleotide and the at least one polynucleotide comprises at least one of a mRNA, a DNA, a siRNA, a gRNA, a chimera, or fragment thereof or combination thereof.

74. A kit comprising the mixture according to any one of claims 50 to 73, and at least one container.

75. A composition comprising a mixture of minimally agglomerated coated therapeutic agent-, antigen-, immunogen-, and / or compound- or combination thereof particles or microparticles, wherein a coated plurality of therapeutic agent-, antigen-, immunogen-, and / or compound- or combination thereof particles or microparticles comprise at least one of a core, one or more coating layers encasing the core, and a plurality of milling agents.

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