Particles comprising therapeutic or diagnostic agent and suspensions and methods of use thereof

Abstract

To provide particles, compositions including the particles, and methods of making the particles using electrospray, wherein the particles enable high concentrations of therapeutic or diagnostic agents to be delivered at low viscosity and the particles also exhibit beneficial properties in terms of stability.SOLUTION: The present invention provides a method of electrospraying a first liquid comprising a first therapeutic or diagnostic agent to form droplets and evaporating the first liquid to produce particles from the droplets, wherein the therapeutic or diagnostic agent in the particles has 0.5 to 1.0 activity per unit.SELECTED DRAWING: None

Description

[Technical Field]

[0001] Background of the Invention There is an urgent and unmet need for therapeutic and diagnostic formulations that are reliable, convenient, and cost-effective to administer. This is particularly true for therapeutic proteins, such as monoclonal antibodies, which are becoming increasingly important in the treatment of a wide range of life-threatening and debilitating diseases. Desirable formulations contain high concentrations of therapeutic or diagnostic agents and possess adequate stability, allowing high doses of the agent to be administered using a minimal volume of formulation. In some cases, this helps reduce the time to deliver the required dose and the pain or discomfort experienced by the patient. In some cases, this can also help reduce the frequency of agent administration. However, such formulation attributes are typically not achievable with aqueous solutions. Highly concentrated aqueous therapeutic or diagnostic agents are often characterized by high fluid dynamic viscosity, which prevents the use of standard injection equipment, while degradation of the active ingredient proceeds at an accelerated rate through one or several pathways. Summary of the Invention [Means for solving the problem]

[0002] SUMMARY OF THE INVENTION In one aspect, the present invention provides a method of forming particles by electrospraying, e.g., conventional electrospraying, a stream of a first liquid comprising a first therapeutic or diagnostic agent towards a collector (e.g., another liquid), wherein the particles are collected in the collector, and the concentration of the first therapeutic or diagnostic agent in the liquid is between 0.0001 and 1000 mg / mL, e.g., between 1 and 1000 mg / mL, 1 and 900 mg / mL, The concentration of the encapsulating agent is in the range of 1 to 500 mg / mL, 1 to 250 mg / mL, 1 to 100 mg / mL, 1 to 50 mg / mL, 5 to 1000 mg / mL, 100 to 900 mg / mL, 150 to 800 mg / mL, or 200 to 700 mg / mL, and the viscosity of the liquid is in the range of 0.1 to 5000 cP, e.g., 0.75 to 1.5 cP, 0.1 to 1000 cP, 0.1 to 100 cP, 1 to 5000 cP, 10 to 1000 cP, or 100 to 500 cP. In some embodiments, the present invention provides a method of forming particles by electrospraying an annular flow of an encapsulating material in a second liquid toward a collector and electrospraying a flow of a first liquid centrally relative to the annular flow of the encapsulating material. In some embodiments, the encapsulating material is in the first liquid.

[0003] In another aspect, the invention provides a method of electrospraying a first liquid containing a first therapeutic or diagnostic agent, e.g., by conventional electrospraying, to form droplets, and removing (e.g., evaporating) the first liquid to produce particles from the droplets, wherein the therapeutic or diagnostic agent in the particles has an activity of 0.5 to 1.0 activity / unit, e.g., 0.75 to 1.0 activity / unit, or 0.9 to 1.0 activity / unit.

[0004] In either method, the method may further comprise suspending the particles in a pharmaceutically acceptable vehicle, thereby forming a pharmaceutical composition. Alternatively, the particles may be formulated as a pharmaceutical composition in dry form, e.g., a powder.

[0005] In some embodiments, the encapsulating material comprises poly(vinyl alcohol), poly(acrylic acid), poly(acrylamide), poly(ethylene oxide), poly(lactic acid), poly(glycolic acid), polycaprolactone, poly(lactic-co-glycolic acid), chitosan, cellulose, or any combination thereof. In some embodiments, the encapsulating material is any additive, therapeutic agent, or diagnostic agent.

[0006] In another embodiment, the first liquid is aqueous, an organic solvent, an ionic liquid, a hydrogel, an ionogel, or a combination thereof. The organic solvent may be benzyl alcohol, benzyl benzoate, castor oil, coconut oil, corn oil, cottonseed oil, fish oil, grapeseed oil, hazelnut oil, hydrogenated palm kernel oil, olive oil, peanut oil, peppermint oil, safflower oil, sesame oil, soybean oil, sunflower oil, vegetable oil, walnut oil, polyethylene glycol, glycofurol, acetone, diglyme, dimethylacetamide, dimethyl isosorbide, dimethyl sulfoxide, ethanol, ethyl acetate, ethyl ether, ethyl lactate, isopropyl acetate, methyl acetate, methyl isobutyl ketone, methyl tert-butyl ether, N-methylpyrrolidone, perfluorodecalin, 2- Examples of suitable aqueous solutions include pyrrolidone, triglycerides, tetrahydrofurfuryl alcohol, fractionated C8 and C10 vegetable fatty acid triglycerides (e.g., MIGLYOL® 810 and MIGLOYL® 812N), propylene glycol diesters of saturated C8 and C10 vegetable fatty acids (e.g., MIGLYOL® 840), ethyl oleate, ethyl caprate, dibutyl adipate, fatty acid esters, hexanoic acid, octanoic acid, triacetin, diethyl glycol monoether, gamma-butyrolactone, eugenol, clove bud oil, citral, limonene, and any combination thereof. Aqueous liquids include water, 0.9% saline, lactated Ringer's solution, and buffers (e.g., acetate buffer, histidine buffer, succinate buffer, HEPES buffer, Tris buffer, carbonate buffer, citrate buffer, phosphate buffer, glycine buffer, barbital buffer, and cacodylate buffer). The liquid may further comprise other components such as carbohydrates, pH adjusters, salts, chelating agents, minerals, polymers, surfactants, protein stabilizers, emulsifiers, preservatives, amino acids, antioxidants, proteins, organic solvents, or nutrient media. In some embodiments, each of the other components independently represents 0.0001-99% (w / v) of the liquid to be sprayed, e.g., 0.0001-90% (w / v), 0.0001-50% (w / v), 0.0001-10% (w / v), 0.0001-1% (w / v), or 0.0001-0.1% (w / v).Those skilled in the art will be able to determine the appropriate amounts of other ingredients in the liquid to be sprayed. Carbohydrates include dextran, trehalose, sucrose, agarose, mannitol, lactose, sorbitol, and maltose. pH adjusters can be, for example, acetate, citrate, glutamate, glycinate, histidine, lactate, maleate, phosphate, succinate, tartrate, bicarbonate, aluminum hydroxide, phosphoric acid, DL-lactic acid / glycolic acid, phosphorylethanolamine, tromethamine, imidazole, glycylglycine, or monosodium glutamate. Salts include sodium chloride, calcium chloride, potassium chloride, sodium hydroxide, stannous chloride, magnesium sulfate, sodium glucoheptonate, sodium pertechnetate, or guanidine hydrochloride. Chelating agents can be, for example, disodium edetate. Minerals can be, for example, calcium, zinc, or titanium dioxide. Suitable polymers include propylene glycol, glucose star polymers, silicone polymers, polydimethylsiloxane, polyethylene glycol, carboxymethylcellulose, poly(glycolic acid), poly(lactic-co-glycolic acid), and polylactic acid. Surfactants can be, for example, polysorbates, magnesium stearate, sodium dodecyl sulfate, polyethylene glycol nonylphenyl ether (Triton™ N-101), glycerin, or polyoxyethylated castor oil. Protein stabilizers include acetyltryptophan salts, caprylate salts, and N-acetyltryptophan. Emulsifiers can be, for example, polysorbate 80, polysorbate 20, sorbitan monooleate, ethanolamine, polyoxyl 35 castor oil, polyoxyl 40 hydrogenated castor oil, carbomer 1342, corn oil mono-di-triglyceride, polyoxyethylated oleic acid glyceride, or poloxamer. Preservatives include phenol, m-cresol, benzyl alcohol, 2-phenyloxyethanol, chlorobutanol, neomycin, benzethonium chloride, glutaraldehyde, or beta-propiolactone.The amino acid may be, for example, alanine, aspartic acid, cysteine, isoleucine, glutamic acid, leucine, methionine, phenylalanine, pyrrolidine, serine, selenocysteine, threonine, tryptophan, tyrosine, valine, asparagine, L-arginine, histidine, glycine, or glutamine. The antioxidant may be, for example, glutathione, ascorbic acid, cysteine, or tocopherol. The protein may be, for example, protamine, protamine sulfate, or gelatin. The organic solvent may be dimethyl sulfoxide, N-methyl-pyrrolidone, N-ethyl-pyrrolidone, or a mixture thereof. Suitable preservatives include methyl hydroxybenzoate, thimerosal, parabens, formaldehyde, and castor oil. The liquid may further include adenine, tri-n-butyl phosphate, octafluoropropane, white petrolatum, or p-aminophenyl-p-anisate. Exemplary ionic liquids include pyridinium, pyridazinium, pyrimidinium, pyrazinium, imidazolium, pyrazolium, thiazolium, oxazolium, triazolium, ammonium, sulfonium, halides, sulfates, sulfonates, carbonates, phosphates, bicarbonates, nitrates, acetates, and PF6. - , BF4 - , triflate, nonaflate, bis(trifyl)amide, trifluoroacetate, heptafluorobutanoate, haloaluminate, or any combination thereof. Exemplary hydrogels or ionogels are collagen hydrogels, chitosan hydrogels, methylcellulose hydrogels, dextran hydrogels, alginate hydrogels, agarose hydrogels, poly(methyl methacrylate) hydrogels, poly(amidoamine) hydrogels, poly(ethyleneimine) hydrogels, polyethylene oxide hydrogels, gelatin hydrogels, hyaluronic acid hydrogels, and any combination thereof.

[0007] In some embodiments, the pharmaceutical composition has a concentration of the first therapeutic or diagnostic agent of 0.0001 to 1000 mg / mL, for example, 100 to 800, 200 to 700, 200 to 600, or 300 to 700 mg / mL.

[0008] In some embodiments, the pharmaceutical composition includes a second therapeutic or diagnostic agent, for example, at a concentration of 0.0001 to 1000 mg / mL. The first therapeutic or diagnostic agent and the second therapeutic or diagnostic agent can be the same or different.

[0009] Therapeutic and diagnostic agents include nucleic acids, oligonucleotides, antibodies, amino acids, peptides, proteins, cells, bacteria, gene therapy drugs, genome-engineered therapeutic drugs, epigenome-engineered therapeutic drugs, carbohydrates, chemical drugs, contrast agents, magnetic particles, polymer beads, metal nanoparticles, metal microparticles, quantum dots, antioxidants, antibiotics, hormones, nucleoproteins, polysaccharides, glycoproteins, lipoproteins, steroids, analgesics, local anesthetics, anti-inflammatory agents, antimicrobial agents, chemotherapeutic agents, exosomes, outer membrane vesicles, vaccines, viruses, bacteriophages, adjuvants, vitamins, minerals, organelles, and any combination thereof.

[0010] In some embodiments, either the electrosprayed second liquid, the liquid collector, or the suspension medium is aqueous, an organic solvent, an ionic liquid, a hydrogel, an ionogel, or a combination thereof. Organic solvents for use in the medium include benzyl alcohol, benzyl benzoate, castor oil, coconut oil, corn oil, cottonseed oil, fish oil, grapeseed oil, hazelnut oil, hydrogenated palm kernel oil, olive oil, peanut oil, peppermint oil, safflower oil, sesame oil, soybean oil, sunflower oil, vegetable oil, walnut oil, polyethylene glycol, glycofurol, acetone, diglyme, dimethylacetamide, dimethyl isosorbide, dimethyl sulfoxide, ethanol, ethyl acetate, ethyl ether, ethyl lactate, isopropyl acetate, methyl acetate, methyl isobutyl ketone, methyl tert-butyl ether, N-methylpyrrolidone, perfluorodecamethylolpropane ... phosphorus, 2-pyrrolidone, triglycerides, tetrahydrofurfuryl alcohol, fractionated triglycerides of C8 and C10 vegetable fatty acids (e.g., MIGLYOL® 810 and MIGLOYL® 812N), propylene glycol diesters of saturated C8 and C10 vegetable fatty acids (e.g., MIGLYOL® 840), ethyl oleate, ethyl caprate, dibutyl adipate, fatty acid esters, hexanoic acid, octanoic acid, triacetin, diethyl glycol monoether, gamma-butyrolactone, eugenol, clove bud oil, citral, limonene, and any combination thereof. Exemplary aqueous liquids are water, 0.9% saline, lactated Ringer's solution, and buffers (e.g., acetate buffer, histidine buffer, succinate buffer, HEPES buffer, Tris buffer, carbonate buffer, citrate buffer, phosphate buffer, glycine buffer, barbital buffer, and cacodylate buffer). The medium may further comprise additional components such as carbohydrates, pH adjusters, salts, chelating agents, minerals, polymers, surfactants, protein stabilizers, emulsifiers, preservatives, amino acids, antioxidants, proteins, organic solvents, or nutrient media.In some embodiments, each of the other components independently constitutes 0.0001-99% (w / v) of the liquid, e.g., 0.0001-90% (w / v), 0.0001-50% (w / v), 0.0001-10% (w / v), 0.0001-1% (w / v), or 0.0001-0.1% (w / v). One of skill in the art would be able to determine appropriate amounts of the other components in the liquid. Carbohydrates include dextran, trehalose, sucrose, agarose, mannitol, lactose, sorbitol, or maltose. pH adjusters include acetate, citrate, glutamate, glycinate, histidine, lactate, maleate, phosphate, succinate, tartrate, bicarbonate, aluminum hydroxide, phosphoric acid, hydrochloric acid, DL-lactic acid / glycolic acid, phosphorylethanolamine, tromethamine, imidazole, glycylglycine, or monosodium glutamate. Salts include sodium chloride, calcium chloride, potassium chloride, sodium hydroxide, stannous chloride, magnesium sulfate, sodium glucoheptonate, sodium pertechnetate, or guanidine hydrochloride. An exemplary chelating agent is disodium edetate. Minerals include calcium, zinc, and titanium dioxide. Polymers include propylene glycol, glucose star polymers, silicone polymers, polydimethylsiloxane, polyethylene glycol, carboxymethylcellulose, poly(glycolic acid), poly(lactic acid-co-glycolic acid), and polylactic acid. Surfactants include polysorbates, magnesium stearate, sodium dodecyl sulfate, polyethylene glycol nonylphenyl ether (Triton™ N-101), glycerin, or polyoxyethylated castor oil. Protein stabilizers include acetyltryptophan salts, caprylate salts, or N-acetyltryptophan. Emulsifiers can be, for example, polysorbate 80, polysorbate 20, sorbitan monooleate, ethanolamine, polyoxyl 35 castor oil, polyoxyl 40 hydrogenated castor oil, carbomer 1342, corn oil mono-di-triglyceride, polyoxyethylated oleic acid glyceride, or poloxamer.Preservatives include phenol, m-cresol, benzyl alcohol, 2-phenyloxyethanol, chlorobutanol, neomycin, benzethonium chloride, glutaraldehyde, and beta-propiolactone. The amino acid may be, for example, alanine, aspartic acid, cysteine, isoleucine, glutamic acid, leucine, methionine, phenylalanine, pyrrolidine, serine, selenocysteine, threonine, tryptophan, tyrosine, valine, asparagine, L-arginine, histidine, glycine, or glutamine, for example, asparagine, L-arginine, histidine, glycine, or glutamine. Suitable antioxidants include glutathione, ascorbic acid, cysteine, and tocopherol. The protein may be protamine, protamine sulfate, or gelatin. The organic solvent can be dimethyl sulfoxide, N-ethyl-pyrrolidone, N-methyl-pyrrolidone, or a mixture thereof. The preservative can be, for example, methyl hydroxybenzoate, thimerosal, paraben, formaldehyde, or castor oil. The medium can further include, for example, adenine, tri-n-butyl phosphate, octafluoropropane, white petrolatum, or p-aminophenyl-p-anisate. The ionic liquid can be, for example, pyridinium, pyridazinium, pyrimidinium, pyrazinium, imidazolium, pyrazolium, thiazolium, oxazolium, triazolium, ammonium, sulfonium, halide, sulfate, sulfonate, carbonate, phosphate, bicarbonate, nitrate, acetate, or PF6. - , BF4 - , triflate, nonaflate, bis(trifyl)amide, trifluoroacetate, heptafluorobutanoate, haloaluminate, or any combination thereof. Exemplary hydrogels or ionogels are collagen hydrogels, chitosan hydrogels, methylcellulose hydrogels, dextran hydrogels, alginate hydrogels, agarose hydrogels, poly(methyl methacrylate) hydrogels, poly(amidoamine) hydrogels, poly(ethyleneimine) hydrogels, polyethylene oxide hydrogels, gelatin hydrogels, hyaluronic acid hydrogels, and any combination thereof.

[0011] In some embodiments, the amount of additional compound, i.e., additive, present in the first liquid, second liquid, recovery liquid, or medium is as shown in the table below. The percentages are as a mass percentage of the total solute loading. For a first liquid containing a therapeutic agent at a concentration of 10 mg / mL and an additive at a concentration of 5 mg / mL, for example, the mass fraction of the additive relative to the total solute population is 33%.

[0012] [Table 1]

[0013] For the second liquid, recovery liquid, and vehicle, Range 4 can be up to 100% for any additive listed in the table.

[0014] In some embodiments, the particles have a diameter of 0.1 to 1000 μm, e.g., 1 to 400 μm, 1 to 200 μm, 1 to 100 μm, 1 to 50 μm, 1 to 25 μm, 1 to 10 μm, 10 to 100 μm, 50 to 100 μm, 50 to 75 μm, or 75 to 100 μm.

[0015] In one embodiment, the particles have a polydispersity index of 0.05 to 0.9.

[0016] In some embodiments, the pharmaceutical composition has a viscosity of 0.27 to 200 cP, for example, 0.27 to 100 cP, 0.27 to 50 cP, 0.27 to 30 cP, 20 to 50 cP, 1 to 30 cP, 1 to 20 cP, or 1 to 15 cP.

[0017] In some embodiments, the pharmaceutical composition comprises 5 to 90% by volume of particles, for example, 20 to 90% by volume, 40 to 80% by volume, 50 to 60% by volume, or 70 to 90% by volume of particles.

[0018] In a related aspect, the invention provides a composition comprising particles made by the methods of the invention.

[0019] In another aspect, the invention provides a method of administering a first therapeutic or diagnostic agent by administering a pharmaceutical composition comprising particles produced by the methods of the invention.

[0020] In another aspect, the present invention provides a method for administering a first therapeutic or diagnostic agent to a mammal. The method comprises administering an effective amount of a pharmaceutical composition to the mammal. In one embodiment, the pharmaceutical composition comprises a vehicle and particles, the particles comprising the therapeutic or diagnostic agent, wherein the pharmaceutical composition has a viscosity of 0.27 to 200 cP, e.g., 0.27 to 100 cP, 0.27 to 50 cP, 0.27 to 30 cP, 20 to 50 cP, 1 to 30 cP, 1 to 20 cP, or 1 to 15 cP, and a concentration of the first therapeutic or diagnostic agent of 0.0001 to 1000 mg / mL, e.g., 100 to 800, 200 to 700, 200 to 600, or 300 to 700 mg / mL. In another embodiment, the pharmaceutical composition is in dry form particles comprising the therapeutic or diagnostic agent. In a related aspect, the present invention provides a composition, e.g., a pharmaceutical composition, comprising particles comprising a first therapeutic or diagnostic agent. In some embodiments, the composition further comprises a vehicle, wherein the composition has a viscosity of 0.27-200 cP, e.g., 0.27-100 cP, 0.27-50 cP, 0.27-30 cP, 20-50 cP, 1-30 cP, 1-20 cP, or 1-15 cP, and a concentration of the first therapeutic or diagnostic agent of 0.0001-1000 mg / mL, e.g., 100-800, 200-700, 200-600, or 300-700 mg / mL. In other embodiments, the composition comprises the particles in dry form.

[0021] In some embodiments, the composition comprises 5-90% by volume of particles, for example, 20-90%, 40-80%, 50-60%, or 70-90% by volume of particles.

[0022] Administration may be via the auricular, buccal, conjunctival, cutaneous, dental, electroosmotic, intracervical, intrasinus, intratracheal, enteral, epidural, extraamniotic, extracorporeal, infiltrative, interstitial, intraperitoneal, intraamniotic, intra-arterial, intra-articular, intrabiliary, intra-bronchial, intra-vesical, intracardiac, intracartilaginous, intrasacral, intracavity, intracavity, intracerebral, intracisternal, intracorneal, intracoronary, intracoronary, intracavernosal, intradermal, intradiscal, intraductal, intraduodenal, intradural, intraepidermal, intraesophageal, intragastric, intragingival, intraileal, intralesional, intraluminal, intralymphatic, intramedullary, intrameningeal, intramuscular, intraocular, intraovarian, intrapericardial, intraperitoneal, intrapleural, intraprostatic, intrapulmonary, or nasal routes. Administration may be by intrasinus or periorbital, intraspinal, intrasynovial, intratendinous, intratesticular, intrathecal, intrathoracic, intratubular, intratumoral, intratympanic, intrauterine, intravascular, intravenous, intravenous bolus, intravenous infusion, intraventricular, intravesical, intravitreal, iontophoretic, lavage, laryngeal, nasal, nasogastric, occlusive dressing, ocular, oral, oropharyngeal, parenteral, transdermal, periarticular, epidural, perineural, periodontal, rectal, inhalation, retrobulbar, soft tissue, subarachnoid, subconjunctival, subcutaneous, sublingual, submucosal, topical, transdermal, transmucosal, transplacental, transtracheal, transtympanic, ureteral, urethral, ​​or vaginal administration.

[0023] In some embodiments of any aspect of the invention, the viscosity is measured at shear rates in the Newtonian region. -1 More than, say, 1000s -1 of or 1000s -1 Measured at higher shear rates.

[0024] In some embodiments, the concentration of the first therapeutic or diagnostic agent in the first liquid is 0.0001 to 1000 mg / mL, for example, 1 to 1000 mg / mL, 1 to 900 mg / mL, 1 to 500 mg / mL, 1 to 250 mg / mL, 1 to 100 mg / mL, 1 to 50 mg / mL, 5 to 1000 mg / mL, 100 to 900 mg / mL, 150 to 800 mg / mL, or 200 to 700 mg / mL.

[0025] In another aspect, the present invention provides a method of administering a first therapeutic or diagnostic agent to a mammal. The method comprises administering to the mammal an effective amount of a suspension or dry formulation of particles comprising the therapeutic or diagnostic agent, wherein the first therapeutic or diagnostic agent has an activity of 0.5 to 1.0 activity / unit, e.g., 0.75 to 1.0 activity / unit, or 0.9 to 1.0 activity / unit (e.g., about 0.99 activity / unit). In a related aspect, the present invention provides a composition comprising particles comprising a first therapeutic or diagnostic agent having an activity of 0.5 to 1.0 activity / unit, e.g., 0.75 to 1.0 activity / unit, or 0.9 to 1.0 activity / unit. The composition may be a suspension of particles in a non-aqueous or aqueous liquid. Alternatively, the composition may be in a dry form, e.g., a powder, for inhalation or needleless injection, etc. The composition may be in the form of a pharmaceutical composition in which the first therapeutic or diagnostic agent is present in an effective amount.

[0026] In some embodiments, the suspension has a viscosity of 0.27 to 200 cP, for example, 0.27 to 100 cP, 0.27 to 50 cP, 0.27 to 30 cP, 20 to 50 cP, 1 to 30 cP, 1 to 20 cP, or 1 to 15 cP.

[0027] In some embodiments, the suspension comprises 5-90% by volume of particles, such as, for example, 20-90% by volume, 40-80% by volume, 50-60% by volume, or 70-90% by volume of particles.

[0028] In some embodiments, the suspension has a concentration of the first therapeutic or diagnostic agent of 0.0001 to 1000 mg / mL, for example, 100 to 800, 200 to 700, 200 to 600, or 300 to 700 mg / mL.

[0029] In some embodiments, the liquid or dry formulation in suspension includes a second therapeutic or diagnostic agent, for example, at a concentration of 0.0001-1000 mg / mL. The first therapeutic or diagnostic agent and the second therapeutic or diagnostic agent can be the same or different.

[0030] definition

[0031] The term "activity" refers to the ratio of a functional or structural aspect of a therapeutic or diagnostic agent at two time points. The denominator of the ratio corresponds to a measure of the functional or structural aspect of the therapeutic or diagnostic agent in the feed solution immediately prior to electrospray particle formation. The numerator of the ratio corresponds to the same measure of the functional or structural aspect of the therapeutic or diagnostic agent at a later time point, e.g., immediately after electrospray particle formation. In certain embodiments, protein activity is assessed by SEC-HPLC or the protein's propensity to bind to a selected target.

[0032] The term "conventional electrospray" is used to refer to electrospray generated with an assembly including a capillary tube, a hydraulic pump, a power supply, and a counter electrode. The capillary tube and counter electrode are spaced a distance apart and disposed in a dielectric medium. The power supply electrically couples the tube to the counter electrode, thereby creating a relative electrical bias. This generates an electric field whose action drives electrospray when a pump is used to eject liquid from the end of the capillary tube.

[0033] The "dry" particle component, i.e., the dry core or dry shell containing the therapeutic or diagnostic agent, has undergone a drying step or a series of drying steps. As a result, its moisture or solvent content is substantially reduced compared to that before drying. In some embodiments, the residual moisture or solvent content of the dry component is less than about 10% by weight, e.g., less than about 5% by weight. Exemplary methods for measuring moisture content include chemical titration, e.g., Karl Fischer titration with a vacuum oven. Various solvents, including water, can also be measured using weight loss methods with thermal excitation. Exemplary methods include thermogravimetric analysis with infrared spectroscopy (TGA-IR).

[0034] The term "electrospray" refers to a process in which droplets of a first liquid are formed in a dielectric medium in the presence of an electric field. Suitable dielectric media include vacuum, air, and a second liquid that is at least partially immiscible with the first liquid. The droplets need not be electrically charged (e.g., droplets can be formed from an electrically insulating liquid such as oil), and an electric field need not be the primary driving force behind droplet formation. In some embodiments, the droplets are generated by conventional electrospray (M. Cloupeau and B. Prunet-Foch, J. Aerosol Sci., vol. 25, no. 6, pp. 1021-1036, 1994; J.F. de la Mora, Annu. Rev. Fluid Mech., vol. 39: 217-243, 2007). In other embodiments, droplets are formed under an electric field by a sprayer, such as an air nozzle sprayer (Z. Takats, J.M. Wiseman, B. Gologan, and R.G. Cooks, Anal. Chem., 2004, 76 (14), pp. 4050-4058). In other embodiments, droplets are formed under an electric field by a microfluidic device, such as a T-junction (H. Kim, D. Luo, D. Link, D. Weitz, M. Marquez, and Z. Cheng, Appl. Phys. Lett., 91, 133106, 2007). In some embodiments, electrospray is a gentle method of spraying used to generate particles. In some embodiments, in electrospraying, a liquid or gel (or pre-gel) is pumped from a supply through a capillary nozzle into an electric field formed at the nozzle opening, which causes the liquid or gel (or pre-gel) to disperse away from the nozzle as a uniform particle aerosol.

[0035] The term "electrospray particle formation" refers to a process in which a first liquid containing a solute is electrosprayed to form droplets, and then the first liquid is removed to form particles containing the solute. Removal of the first liquid, or drying, can be performed by one of several methods known in the art, or any combination thereof. In some embodiments, drying is performed by contacting the droplets with a hot gas stream, such as in spray drying (B. Gikanga, R. Turok, A. Hui, M. Bowen, O.B. Stauch, Y. Maa, J. Pharm Sci. Tech., 2015, 69, 59-73). In other embodiments, drying is performed by freezing the droplets and then sublimating the first liquid, such as in spray freeze drying (S. Wanning, R. Suverkrup, A. Lamprecht, Int. J. Pharm., 2015, 488, 136-153). In other embodiments, drying is performed, such as in certain microfluidic systems, by contacting a first liquid with a second liquid that is at least partially miscible with it, thereby precipitating the solute (Aniket, DA Gaul, DL Rickard, D. Needham J. Pharm. Sci., 2014, 103, 810-820).

[0036] The term "encapsulating material" refers to a material that can be dried or gelled around a particle core to form a shell.

[0037] The term "additive" refers to an addition to a formulation, the effect of which may be useful for achieving desired modifications to the properties of the formulation or formulation. Such modifications include, but are not limited to, physical stability, chemical stability, and therapeutic efficacy. Exemplary additives include, but are not limited to, carbohydrates, pH adjusters, salts, chelating agents, minerals, polymers, surfactants, protein stabilizers, emulsifiers, preservatives, amino acids, antioxidants, proteins, organic solvents, or nutrient media.

[0038] The term "feed solution" refers to a preparation of a therapeutic or diagnostic agent in a first liquid as a solution, slurry, or some other liquid form. In some embodiments, the preparation contains additives and, optionally, a buffer.

[0039] The term "injectability" refers to the relative ease with which a liquid formulation can be administered to a subject via use of an injection device. In some embodiments, injectability is determined by measuring the viscosity of the formulation at various shear rates. In some embodiments, injectability is determined by measuring the sliding yield stress and / or sliding equilibrium stress required to actuate a standard injection device consisting of a syringe barrel, plunger, and, optionally, a needle. In some embodiments, the injectability of a suspension formulation is superior to that of an aqueous formulation containing approximately the same concentration of a therapeutic or diagnostic agent.

[0040] The term "injection breakaway force" refers to the force required to overcome the friction between the syringe barrel and plunger of a standard injection device before the contents of the syringe can be expelled at a consistent rate. This force is applied to the outward-facing end of the syringe plunger shaft and directed along the axis of the syringe barrel. The contents of the syringe are optionally expelled through a syringe needle of a predetermined gauge and length. In some embodiments, the injection breakaway force is measured through a load cell placed on the outward-facing end of the syringe plunger during actuation.

[0041] The term "injection glide force" refers to the force required to maintain consistent delivery of the contents of a standard injection device. This force is applied to the outward-facing end of the syringe plunger shaft and is directed along the axis of the syringe barrel. The contents of the syringe are optionally delivered through the tip of a syringe needle of a predetermined gauge and length. In some embodiments, the injection glide force is measured through a load cell located on the outward-facing end of the syringe plunger during actuation.

[0042] The term "medium" refers to the liquid in which the particles are dispersed.

[0043] The term "Newtonian region" refers to the range of shear rates over which the maximum and minimum viscosity values ​​differ by at most 1% of the maximum value.

[0044] The term "particle" refers to a quantity of therapeutic or diagnostic material that, in one aspect, is in a state of matter that is substantially solid or in gel form compared to a liquid droplet. In some embodiments, the particle comprises a core and a shell, where the shell is considered to be the encapsulating material. In other embodiments, the particle does not comprise a shell, in which case the particle is composed entirely of the core.

[0045] The term "pharmaceutical composition" refers to a composition in which a therapeutic or diagnostic agent retains or partially retains its intended biological activity or functional form and contains only pharmaceutically acceptable components.

[0046] A "pharmaceutically acceptable" component, eg, an excipient, is a component that is suitable for administration to a subject, eg, a human.

[0047] The term "powder formulation" refers to a solid formulation comprising solid particles in the absence of a carrier liquid. In some embodiments, the powder formulation is suitable for powder injection, for example, using a Portal PRIME device.

[0048] The term "Rayleigh limit" refers to the specific charge, e.g., in units of coulombs per kilogram, corresponding to the point where repulsive Coulombic forces overcome the cohesive forces of surface tension of the droplet, resulting in Coulombic breakup.

[0049] The term "stabilizer" refers to an additive or mixture of additives that stabilizes the physical and / or chemical properties of a pharmaceutical formulation. In some embodiments, the stabilizer prevents degradation of the therapeutic or diagnostic agent, for example, during electrospraying, drying, and / or storage of the particulate matter. Exemplary stabilizers include, but are not limited to, sugars, salts, hydrophobic salts, surfactants, reducing agents, cyclodextrins, polyols, carboxylic acids, and amino acids.

[0050] A "stable" formulation refers to a formulation in which a therapeutic or diagnostic agent retains an acceptable portion of its essential physical and / or chemical and / or biological properties over an acceptable period of time. For proteins and peptides, for example, exemplary methods for assessing stability are reviewed in (i) Peptide and Protein Drug Delivery, 247-301, Vincent Lee Ed., Marcel Dekker, Inc., New York, NY, 1991, and (ii) Jones, A., Adv. Drug Delivery Rev. 10: 29-90 (1993). In some embodiments, the chemical stability of a protein is assessed by measuring the size distribution of samples at several stages. These include, for example, before particle formation (evaluation of the feed solution), immediately after particle formation, and even after a period of storage (where storage occurs in or without a suspension formulation carrier medium). In some embodiments, the size distribution is assessed by size exclusion chromatography (SEC-HPLC).

[0051] A "sterile" preparation is aseptic or free of living microorganisms and their spores.

[0052] The term "suspension formulation" refers to a liquid formulation containing solid particles disposed within a carrier liquid that do not dissolve within a reasonable timescale. The particles may settle over time; that is, the physical stability of the suspension is not indefinite, but it can be resuspended using some form of agitation or shaking.

[0053] "Therapeutic amount" refers to the amount of a therapeutic or diagnostic agent needed to produce a desired effect.

[0054] As used herein, the terms "treat," "treated," and "treating" refer to both therapeutic treatment and prophylactic or preventative measures, the purpose of which is to prevent or slow (alleviate) an undesirable physiological condition, disorder, or disease, or to obtain a beneficial or desired clinical result. Beneficial or desired clinical results include, but are not limited to, alleviation of symptoms; a diminution in the extent of the condition, disorder, or disease; a stable (i.e., not worsening) state of the condition, disorder, or disease; a delay in the onset or slowing of the condition, disorder, or disease progression; an improvement or remission (whether partial or total) of the condition, disorder, or disease state, whether detectable or undetectable; an improvement in at least one measurable physical parameter, not necessarily discernible by the patient; or an enhancement or amelioration of the condition, disorder, or disease. Treatment includes eliciting a clinically significant response without excessive levels of side effects. Treatment also includes prolonging survival compared to expected survival if not receiving treatment.

[0055] The term "viscosity" is used to describe the property of a fluid that acts to resist shear flow. For purposes of the present invention, viscosity can be determined at a specific shear rate at 25°C using a rheometer, for example, an AR-G2 rheometer (TA Instruments, USA) equipped with a cone and plate (2° / 40 mm). In some embodiments, viscosity is measured at a shear rate in the Newtonian region. In other embodiments, viscosity is measured at a shear rate of 100 s -1 More than, say, 1000s-1 or 1000s -1 Measured at higher shear rates. [Brief explanation of the drawings]

[0056] BRIEF DESCRIPTION OF THE DRAWINGS

[0057] [Figure 1] Figure 1 shows a basic electrospray assembly. One end of a tube 3 is positioned a distance from an electrode 4, and the other end is attached to a syringe 1 controlled by a syringe pump 2. A power supply 5 charges the tube 3 relative to the electrode 4, creating an electric field 6 in the region between the two.

[0058] [Figure 2] 2 shows a basic electrospray assembly, in which a reservoir 7 containing a liquid 8 is placed between the end of a tube 3 and an electrode 4. The end of the tube 3 is not immersed in the liquid 8.

[0059] [Figure 3] 3 shows a basic electrospray assembly in which a reservoir 7 containing a liquid 8 is placed between the end of a tube 3 and an electrode 4. The end of the tube 3 is immersed in the liquid 8.

[0060] [Figure 4] Figure 4 shows a coaxial electrospray assembly. Tubing 3 is connected to syringe 1, which is controlled by syringe pump 2. A second tube 11 is connected to a second syringe 9, which is controlled by syringe pump 10. The distal ends of tubes 3 and 11 are connected to adapter 12, which outputs coaxial tubing 13, the end of which is spaced apart from electrode 4. Power source 5 charges coaxial tubing 13 relative to electrode 4, creating an electric field 6 in the region between the two.

[0061] [Figure 5]5 shows a coaxial electrospray assembly in which a reservoir 7 containing a liquid 8 is positioned between the end of a tube 13 and an electrode 4. The end of the tube 13 is not immersed in the liquid 8.

[0062] [Figure 6] 6 shows a coaxial electrospray assembly in which a reservoir 7 containing a liquid 8 is positioned between the end of a tube 13 and an electrode 4. The end of the tube 13 is immersed in the liquid 8.

[0063] [Figure 7] 7 shows the meniscus at the end of the tube when the electric field 6 is below the electrospray threshold. In a conventional single-liquid tube 3, the meniscus of liquid 16 does not form an electrospray jet. In a coaxial assembly including outer and inner tubes 14 and 15, and outer and inner liquids 17 and 16, the meniscus at the end of the assembly does not form a coaxial electrospray jet.

[0064] [Figure 8] 8 shows droplet formation from the end of the tube when the electric field 6 is above the electrospray threshold. In a conventional single-liquid tube 3, the jet 18 breaks up into a droplet collection 20, where droplets 22 contain a single liquid 16. This is accompanied by a flow 25 of liquid 16 through the tube 3. In a coaxial electrospray assembly, the coaxial jet 19 breaks up into a core-shell droplet collection 21 having a core 23 of liquid 16 and a shell 24 of liquid 17. This is accompanied by a flow 26 of liquid 16 through the inner tube 15 and a flow 27 of liquid 17 through the outer tube 14.

[0065] [Figure 9]9 shows a comparison of suspension formulations and solutions produced by the disclosed methods. Solutions 29 of therapeutic or diagnostic agents may be too viscous to be administered using a standard syringe 28 due to troublesome intermolecular interactions 30. In contrast, suspension formulations 31 of particles 32 containing therapeutic or diagnostic agents 33 have a much lower effective viscosity and may be able to be administered using a standard syringe 28.

[0066] [Figure 10] FIG. 10 is a series of images of particles of human IgG protein formed from an aqueous solution by electrospray particle generation.

[0067] [Figure 11] Figure 11 shows an image of bovine serum albumin particles generated by electrospray particle generation with an ImageJ analysis overlay. Scale bar is 50 μm.

[0068] [Figure 12] Figure 12 is a series of images showing human IgG particles generated by electrospray particle generation. Image 37 is an optical image at 20X magnification. Images 38 and 39 are SEM images at 2000X and 5000X magnification, respectively.

[0069] [Figure 13] Figure 13 is a series of optical images of monoclonal antibody particles generated by electrospray particle generation. Scale bar is 50 μm.

[0070] [Figure 14] FIG. 14 is a graph showing the results of an ELISA assay control experiment for human IgG.

[0071] [Figure 15] FIG. 15 is a graph showing a comparison of ELISA signals for untreated (before electrospray particle formation) and electrosprayed human IgG.

[0072] [Figure 16] FIG. 16 is a series of graphs showing cell binding data for an electrosprayed monoclonal antibody (mAb1).

[0073] [Figure 17] FIG. 17 is a series of graphs showing cell binding data for an electrosprayed monoclonal antibody (mAb2).

[0074] [Figure 18] 18 is a series of graphs showing cell binding data for an electrosprayed monoclonal antibody (mAb1) before and after accelerated storage. The particles were stored with and without a suspension medium (carrier).

[0075] [Figure 19] FIG. 19 is a graph showing viscosity data for various aqueous and suspension formulations at different concentrations.

[0076] [Figure 20] Figure 20 is an image showing human IgG particles generated by a nebulizer using an electric field. The scale bar is 10 μm.

[0077] [Figure 21] Figure 21 is an image showing human IgG particles generated by a nebulizer without an electric field. The scale bar is 10 μm.

[0078] [Figure 22] Figure 22 is an image showing a population of human IgG particles with salts protruding from the particle surface. Scale bar is 100 μm.

[0079] [Figure 23] Figure 23 is an image showing a population of human IgG particles with salts protruding from the particle surface. Scale bar is 30 μm.

[0080] [Figure 24] Figure 24 is an image showing a population of human IgG particles with protruding sugars on the surface of the particle. Scale bar is 30 μm.

[0081] [Figure 25] Figure 25 is an image showing a group of human IgG particles with sugars protruding from the surface of the particle. Scale bar is 20 μm.

[0082] [Figure 26] Figure 26 is an image showing a group of monoclonal antibody (mAb3) particles containing sugars protruding from the particle surface. Scale bar is 100 μm.

[0083] [Figure 27] Figure 27 is an image showing a group of monoclonal antibody (mAb3) particles containing sugars protruding from the surface of the particles. Scale bar is 30 μm. DETAILED DESCRIPTION OF THE INVENTION

[0084] Detailed Description The present disclosure provides methods for preparing electrosprayed particles containing one or more therapeutic or diagnostic agents. Particles have traditionally been prepared by milling, conventional spray drying (G. Lee, Spray-Drying of Proteins. In: J.F. Carpenter and M.C. Manning (eds) Rational Design of Stable Protein Formulations, vol. 13. Springer, Boston, MA), freeze-drying, and conventional emulsion techniques (D. Saglam, P. Venema, R. de Vries, L.M.C. Sagis, E. van der Linde, Food Hydrocolloids, 2011, 25, 1139-1148). However, each of these techniques presents challenges for the preparation of particles from certain therapeutic or diagnostic agents. For example, milling proteins requires an initial freeze-drying step and a subsequent milling step, which may affect biological activity. Control over particle morphology is also limited. While spray drying and emulsion techniques can affect the biological function of proteins due to high shear rates, the latter also presupposes high operating temperatures that can be detrimental. A gentler process, e.g., one that preserves biological activity and thereby allows precise control over particle morphology, e.g., one that results in highly monodisperse particles, is that of electrospraying.

[0085] In some embodiments, the particles do not have a shell. In some embodiments, the particles have a liquid, solid, or gel core and a shell. The liquid, solid, or gel contains one or more therapeutic or diagnostic agents. In some embodiments, the therapeutic or diagnostic agent can be in the shell. The therapeutic or diagnostic agent can be dissolved or suspended in the liquid or gel, or otherwise carried by the liquid or gel, e.g., in a slurry. Electrosprayed particles can be incorporated into a vehicle to form a pharmaceutical composition. In some embodiments, the pharmaceutical composition is a highly concentrated colloidal suspension or slurry with a low viscosity, e.g., a viscosity of <50 cP or greater (Dias, C. et al. AAPS PharmSciTech. 2015, 16, 1107), while maintaining stability and a controlled drug release rate of the therapeutic or diagnostic agent. In some embodiments, the present invention allows for the delivery of higher doses of therapeutic or diagnostic agents while minimizing delivery volume, shortening administration time, and / or reducing pain. In another embodiment, a powder composition of a therapeutic or diagnostic agent is provided that can be stored stably for extended periods of time.

[0086] Particles can be formed by electrospraying a first liquid containing a therapeutic or diagnostic agent to form droplets and then removing (e.g., evaporating) the first liquid to generate particles from the droplets. In at least one embodiment, the particles are solid, e.g., they may have a solid shell and a liquid core. The particles can be suspended in a non-aqueous or aqueous liquid, thereby forming a non-aqueous or aqueous suspension. Alternatively, the particles can be used in a dry form, e.g., as a powder. Electrospraying allows for high-throughput, gentle preparation and compatibility with highly viscous feed solutions. Importantly, the process of generating a non-aqueous or aqueous suspension with a therapeutic or diagnostic agent does not significantly alter the structure or biological activity of the agent. Additionally, in some embodiments, the present invention allows for the delivery of higher doses of a therapeutic or diagnostic agent while minimizing delivery volume, shortening administration time, and / or reducing pain.

[0087] Suspension Concept

[0088] In some embodiments, pharmaceutical suspension formulations are formed to improve the injection performance of certain therapeutic or diagnostic agents. Specifically, the suspensions exhibit lower viscosities than aqueous solutions with equivalent therapeutic or diagnostic agent loadings, potentially reducing the force, i.e., sliding yield stress and sliding equilibrium stress, required to administer the suspension using standard syringe equipment. Conceptually, particles in suspension provide a means to replace the intermolecular interactions present in regular solutions, e.g., aqueous solutions, with less troublesome effects, such as excluded volume effects. In some embodiments, this allows the performance of the suspension to approximately follow Einstein's equation for solution viscosity (E.W.J. Mardles, Nature, 1940, 145, 970):

number

[0089] In other embodiments, especially those with high volume fractions φ, the performance of the suspension approximately follows other equations such as the Krieger-Dougherty equation or the Frankel-Acrivos equation, among others (S. Ueller, EW Llewellin, HM Mader, Proc. Royal Soc. A, 2010, 466, 2116).

[0090] In certain embodiments, suspension formulations provide a means to improve the stability of certain therapeutic or diagnostic agents at a given concentration, as compared to, for example, aqueous formulations, while also improving injectability. In other embodiments, where injectability is not necessarily improved, suspension formulations improve the stability characteristics of therapeutic or diagnostic agents at a given concentration. In yet other embodiments, powder formulations improve the stability of certain therapeutic or diagnostic agents, as compared to, for example, aqueous formulations.

[0091] particle

[0092] The particles can have a diameter of 0.1 to 1000 μm. For example, the 0.1 to 90 μm range can be delivered through a 25-gauge needle. Larger particles, for example, in the 90 to 230 μm range, can be useful in conjunction with smaller-gauge needles or other delivery routes or modes. In some embodiments, the lower range of 0.1 to 1 μm is of interest. The particles can have a dispersity index of 0.05 to 0.9. Methods for measuring particle size and distribution include imaging flow cytometry and image analysis of scanning electron micrographs of particles, where the mean spherical radius or diameter is calculated based on the cross-sectional area of ​​the particles projected onto the plane of the image.

[0093] Particles may include both a core and a shell. In some embodiments, particles include a core but not a shell. If no shell is present, the core is a gel core or a dry solid-state core, but if the particle includes a gel shell or a dry solid-state shell, the core may exist in a liquid state. Particle morphology may be approximately spherical, mushroom-shaped, or raisin-shaped, among other potential morphologies, depending on the properties of the electrospray feed solution and drying conditions (FIG. 10). In some embodiments, the particle surface may have wrinkles or indentations.

[0094] In some embodiments, the particles have a density of about 1-6 g / cm 3 Skeletal density, for example, about 1 to 5 g / cm 3 , about 1~3g / cm 3 , about 1~2g / cm 3 , about 1~1.5g / cm 3 , or about 1.1 to 1.4 g / cm 3 Exemplary methods for density measurement include gas displacement pycnometry.

[0095] In some embodiments, the residual amount of the first liquid in the particles after drying is 0-10% by weight, e.g., 0-5% by weight, 0-3% by weight, or 0-1% by weight. Exemplary methods for measuring residual solvent content include Karl Fischer titration and various weight loss methods.

[0096] In some embodiments, the particles may exhibit a porosity of about 0-50%, e.g., about 0-10%, about 0-5%, about 0-1%, about 0-0.5%, about 0-0.1%, or about 0-0.01%. Exemplary pore size measurements include scanning electron microscopy (SEM), transmission electron microscopy (TEM), and confocal laser scanning microscopy analysis. The specific surface area of ​​porous microspheres and nanospheres can also be investigated by nitrogen adsorption / desorption analysis and the Brunauer-Emmett-Teller adsorption model. In embodiments where the pore size is sufficiently large, mercury intrusion porosimetry can be used.

[0097] In some embodiments, the particles have a residual net charge of either polarity, i.e., a net positive charge or a net negative charge. In terms of size, the particles can have a charge of 0 to 10 billion, e.g., 0 to 100 million, 0 to 1 million, 0 to 10,000, or 0 to 100. The magnitude of the charge is the magnitude of the charge carried by an electron, i.e., the elementary charge, 1.6x10 -19 It is defined as a coulomb. Exemplary methods for measuring particle charge include those that involve analyzing the movement of the particles in response to an externally applied electric field. In some cases, this is done while the particles are suspended in an insulating liquid such as oil.

[0098] In certain embodiments, the therapeutic or diagnostic agent has a zeta potential of about -90 to 90 mV; e.g., about -60 to 60 mV, about -40 to 40 mV, about -20 to 20 mV, or about -5 to 5 mV. An exemplary method for measuring zeta potential involves reconstituting the therapeutic or diagnostic agent by dissolving the particles in water and analyzing the solution by electrophoretic light scattering, which is similar to dynamic light scattering (DLS) measurements performed in the presence of a positive or negative electric field.

[0099] In some embodiments, the subvisible particles (SVPs) remaining upon particle reconstitution are present in an amount of about 0-10,000 / mL, e.g., 0-6,000 / mL, 0-1,000 / mL, 0-500 / mL, 0-250 / mL, 0-100 / mL, or 0-10 / mL. An exemplary method for measuring SVPs includes microflow imaging, in which a therapeutic or diagnostic agent is reconstituted and diluted to a concentration of about 1 mg / mL.

[0100] In some embodiments, the particles include a loading of therapeutic or diagnostic agent of 1-100 wt%, e.g., 50-100 wt%, 75-100 wt%, 90-100 wt%, 95-100 wt%, 99-100 wt%, or 99.9-100 wt%. At these loadings, the therapeutic or diagnostic agent retains 0.5-1.0 activity, e.g., 0.75-1.0 activity, 0.9-1.0 activity, 0.95-1.0 activity, 0.99-1.0 activity, or 0.999-1.0 activity during electrospray particle formation. This includes activity retained through primary drying and, optionally, secondary drying.

[0101] In some embodiments, dissolution or reconstitution of the particles results in less than 10% aggregates (e.g., less than 8%, less than 5%, less than 4%, less than 3%, or less than 1%) of the diagnostic or therapeutic agent, e.g., protein, as measured, e.g., by HPLC.

[0102] In some embodiments, the particles are flowable. The Hausner ratio can be from 1.0 to greater than 3.0, such as 1.0-3.0, 1.0-2.0, 1.0-1.70 (e.g., very poor), 1.0-1.59, 1.0-1.35, 1.0-1.25, or 1.0-1.11 (e.g., excellent). An exemplary method for measuring powder flowability includes the tapped density method (Carr RL. Chem. Eng., 1965; 72:163-168). Bulk density can be first obtained by adding a known mass of powder to a graduated cylinder. The density can be calculated as mass / volume. The same sample can then be mechanically tapped until no further volume change is observed. The tapped density can then be calculated as the mass divided by the final volume of the powder. A comparison of tapped density to bulk density can be used as an indicator of a powder's ability to flow. Specifically, the Hausner ratio (the unsubmerged apparent or bulk volume, V0) is expressed as the ratio of the final tapped volume, V fThe Hausner ratio (divided by ) is a measure of the ability of a product to sink and allows for an assessment of the relative importance of particle-particle interactions. These interactions are less pronounced in free-flowing powders. The bulk and tapped densities of such free-flowing powders are close in value, so the Hausner ratio is close to 1.0.

[0103] In some embodiments, the particles have one or more of the following characteristics: a size of 1 to 50 μm; a solid core; a gel or solid shell; and a density of 1 to 1.5 g / cm. 3 a density of 0-3 wt%; a residual solvent content of 0-10%; a porosity of 0-10%; a net charge of 0-1 million charges of either polarity, i.e., positive or negative; a therapeutic or diagnostic component having a zeta potential of -60-60 mV; an SVP of 0-1,000 / mL upon reconstitution; a therapeutic or diagnostic agent loading of 50-100 wt%, with the therapeutic or diagnostic agent activity being 0.9-1.0 upon reconstitution; less than 10% aggregates upon reconstitution; and / or a Hausner ratio of 1.0-1.35, or 1.0-1.11.

[0104] The particles can be stored and formulated in various devices for delivery. In some embodiments, the device is a subcutaneous administration device, such as a pre-filled syringe. In some embodiments, the present invention provides a method for making an article of manufacture, comprising filling a container with a suspension formulation. The container in the article of manufacture may include a syringe (e.g., a pre-filled syringe), an autoinjector, a bottle, a vial (e.g., a dual-chamber vial), and a test tube. The container may hold the suspension formulation, and a label on or associated with the container may indicate how to use it. The article of manufacture may further include other materials desirable from a commercial and user standpoint, such as other buffers, diluents, filters, needles, syringes, and package inserts with instructions for use.

[0105] Particle Core

[0106] The core of each particle typically contains one or more therapeutic or diagnostic agents. The core is a solid, dry core if no shell is present, but may be in a liquid state if the particle contains a gel shell or a solid-state dry shell. If a shell is present, the shell may contain therapeutic or diagnostic agents, while the core does not.

[0107] One or more therapeutic or diagnostic agents may be dissolved or suspended in the electrospray feed solution, the first liquid, prior to particle formation. The concentration of the therapeutic or diagnostic agent in the first liquid can range from 0.0001 to 1000 mg / mL. The liquid can be an aqueous or organic solvent, a hydrogel, an ionogel, or a combination thereof. The liquid can be, for example, water, 0.9% saline, lactated Ringer's solution, 5% dextrose, or a buffer solution. In some embodiments, the buffer is acetate buffer, histidine buffer, succinate buffer, HEPES buffer, Tris buffer, carbonate buffer, citrate buffer, phosphate buffer, glycine buffer, barbital buffer, or cacodylate buffer. Organic solvents, hydrogels, and ionogels are described herein. The liquid can further comprise, for example, carbohydrates, pH adjusters, salts, chelating agents, minerals, polymers, surfactants, protein stabilizers, emulsifiers, preservatives, amino acids, antioxidants, proteins, organic solvents, and / or nutrient media. In some embodiments, each of the other components independently comprises 0.0001-99% (w / v), e.g., 0.0001-90% (w / v), 0.0001-50% (w / v), 0.0001-10% (w / v), 0.0001-1% (w / v), or 0.0001-0.1% (w / v) of the liquid to be sprayed. One of skill in the art would be able to determine the appropriate amounts of other components in the liquid to be sprayed. In some embodiments, the carbohydrate is dextran, trehalose, sucrose, agarose, mannitol, lactose, sorbitol, or maltose. In some embodiments, the pH adjuster is acetate, citrate, glutamate, glycinate, histidine, lactate, maleate, phosphate, succinate, tartrate, bicarbonate, aluminum hydroxide, phosphoric acid, hydrochloric acid, DL-lactic acid / glycolic acid, phosphorylethanolamine, tromethamine, imidazole, glycylglycine, or monosodium glutamate. In some embodiments, the salt is sodium chloride, calcium chloride, potassium chloride, sodium hydroxide, stannous chloride, magnesium sulfate, sodium glucoheptonate, sodium pertechnetate, or guanidine hydrochloride.In some embodiments, the chelating agent is edetate disodium. In some embodiments, the mineral is calcium, zinc, or titanium dioxide. In some embodiments, the polymer is propylene glycol, glucose star polymer, silicone polymer, polydimethylsiloxane, polyethylene glycol, carboxymethylcellulose, poly(glycolic acid), poly(lactic-co-glycolic acid), or polylactic acid. In some embodiments, the surfactant is polysorbate, magnesium stearate, sodium dodecyl sulfate, polyethylene glycol nonylphenyl ether (Triton™ N-101), glycerin, or polyoxyethylated castor oil. In some embodiments, the protein stabilizing agent is acetyltryptophan salt, caprylate salt, or N-acetyltryptophan. In some embodiments, the emulsifier is selected from polysorbate 80, polysorbate 20, sorbitan monooleate, ethanolamine, polyoxyl 35 castor oil, polyoxyl 40 hydrogenated castor oil, carbomer 1342, corn oil-mono-di-triglyceride, polyoxyethylated oleic acid glyceride, or poloxamer. In some embodiments, the preservative is phenol, m-cresol, benzyl alcohol, 2-phenyloxyethanol, chlorobutanol, neomycin, benzethonium chloride, glutaraldehyde, or beta-propiolactone. In some embodiments, the amino acid is alanine, aspartic acid, cysteine, isoleucine, glutamic acid, leucine, methionine, phenylalanine, pyrrolidine, serine, selenocysteine, threonine, tryptophan, tyrosine, valine, asparagine, L-arginine, histidine, glycine, or glutamine, such as asparagine, L-arginine, histidine, glycine, or glutamine. In some embodiments, the antioxidant is glutathione, ascorbic acid, cysteine, or tocopherol. In some embodiments, the protein is protamine, protamine sulfate, or gelatin. In some embodiments, the organic solvent may be dimethyl sulfoxide or N-methyl-2-pyrrolidone.In some embodiments, the preservative is methyl hydroxybenzoate, thimerosal, paraben, formaldehyde, or castor oil. In some embodiments, the liquid can further include adenine, tri-n-butyl phosphate, octafluoropropane, white petrolatum, or p-aminophenyl-p-anisate. In some embodiments, the organic solvent can be dichloromethane, dimethyl sulfoxide, urea, sarcosine, methanol, formic acid, acetic acid, ethyl acetate, acetonitrile, acetone, methyl acetate, diethyl ether, hydrazine, ethyl nitrate, butanol, dimethoxyethane, methyl tert-butyl ether, triethylamine, or any combination thereof.

[0108] Particle Shell

[0109] Generally, any additive is suitable as a shell material. Exemplary additives include, but are not limited to, sugars, salts, and amino acids. Therapeutic agents, diagnostic agents, and biocompatible polymers can also be used to form the shell. This includes small molecule drugs. Non-limiting examples of hydrophilic biocompatible polymers include poly(vinyl alcohol), poly(acrylic acid), poly(acrylamide), poly(ethylene oxide), or copolymers or combinations of any two or more thereof. Hydrophilic polymers may be modified to adjust their properties. Alternatively or additionally, the shell component may include one or more biocompatible hydrophobic polymers. Hydrophobic polymers may be modified to adjust their properties. Non-limiting examples of hydrophobic polymers include polycaprolactam, poly(lactic acid), poly(glycolic acid), polycaprolactone, PLGA, or copolymers or combinations of any two or more thereof. In some embodiments, PLGA (50:50) polymer is used as the shell to encapsulate the antibody just below its solubility limit. Polymers can also be prepared with PLGA in various lactic acid-glycolic acid ratios, and can be copolymers with other polymers, such as chitosan, cellulose, etc.

[0110] In some embodiments, the thickness of the particle shell may range from 0 to 90% of the particle diameter. The shell need not be perfectly formed and uniform for encapsulation. In some embodiments, the interface between the shell and the core is partially mixed, so that there is no clear boundary. Furthermore, as described herein, one or more therapeutic or diagnostic agents can be included in the particle shell. The therapeutic or diagnostic agent can be the same or different from that in the core. The concentration of the therapeutic or diagnostic agent in the shell can range from 0.0001 to 300 mg / mL.

[0111] Core-shell ratio

[0112] In embodiments in which the particles include a shell, a core-shell volume ratio of 1:99% to 99:1% by volume, e.g., about 10:90% by volume, about 90:10% by volume, or about 95:5% by volume, is expected to be most useful. A complete coating is not always necessary for sufficient encapsulation. In certain circumstances, for example, for highly concentrated cores, a thicker shell may be beneficial. The core-shell ratio may be useful for adjusting the release kinetics of one or more therapeutic or diagnostic agents. In certain embodiments, it is advantageous to have a polydisperse system, for example, to reduce the viscosity of a suspension formulation. In this case, various core-shell ratios may be of interest.

[0113] Electrospray particle formation

[0114] Electrospraying is a process in which droplets of a first liquid are formed in a dielectric medium in the presence of an electric field. Exemplary dielectric media include vacuum, air, a second liquid that is a suitable electrical insulator and at least partially immiscible with the first liquid, and combinations thereof. The first liquid need not be electrically conductive, and the droplets need not carry a net charge (e.g., droplets can be formed from an electrically insulating liquid such as oil). Furthermore, the electric field acting on the first liquid need not be the primary driving force behind droplet formation. In some embodiments, droplets are formed primarily due to electrostatic interactions between the first liquid and the electric field, such as in conventional electrospraying. In other embodiments, the electric field acts to assist droplet formation by the primary droplet formation device, playing a secondary role and, in some instances, modifying droplet properties. Devices include rotary atomizers, air nozzle atomizers, ultrasonic nozzle atomizers, sonic nozzles, microfluidic T-junctions, microfluidic Y-junctions, microcapillaries, and the like. In all embodiments, the electric field is generated by applying a potential difference between an electrode disposed in a dielectric medium and the first liquid.

[0115] The potential difference between the location where the droplets are initially generated and the electrode, measured in the dielectric medium separating the electrospray source and the electrode, is 0.001 to 100,000 V, e.g., 1 to 50,000 V, 100 to 25,000 V, or 1,000 to 10,000 V. Depending on the Rayleigh limit, the droplets in this field may be charged, on average, to 0 to 1, e.g., 0.1 to 1.0, 0.2 to 1.0, 0.3 to 1.0, 0.4 to 1.0, or 0.5 to 1.0. In some embodiments, the droplets are charged above the Rayleigh limit, inducing Coulombic splitting. The droplets can be dried by any of several techniques known in the literature and then recovered for use. Exemplary collectors include, but are not limited to, electrode plates (N. Bock, TR Dargaville, MA Woodruff, Prog. Polym. Sci., 2012, 37, 1510-1551), liquid baths (FIGS. 2, 3, 5, 6) (US8939388), electrostatic precipitators (YA Haggag, AM Faheem, Front. Pharmacol., 2015, 6, 140), and cyclones (J. Bogelein, G. Lee, Int. J. Pharm., 2010, 401, 68-71).

[0116] Electrospray technology is perhaps most widely known for its use in biological mass spectrometry (J. Fenn, M. Mann, C. Kai Meng, S. Fu Wong, and C. M. Whitehouse, Science, vol. 246, no. 4926, pp. 64–71, October 1989), where its gentle spraying characteristics have enabled the analysis of high-molecular-weight molecules. A typical electrospray device includes a tube and an electrode, which are aligned so that one end of the tube is positioned a distance from the electrode (Figure 1). The tube carries a liquid stream driven by a positive displacement pump, e.g., a syringe pump, or a source of pressurized gas, e.g., a compressed gas bottle. A power source charges the liquid in the tube relative to the electrode, generating an electric field in the region between the distal end of the tube and the electrode. Charge accumulates in the liquid meniscus at the end of the tube in proportion to the strength of this field (Figure 7). When the field reaches a certain threshold strength, the electrostatic drag force acting on the charges becomes sufficient to overcome the surface tension of the meniscus (Figure 8). In some embodiments, this results in the reconstitution of a morphologically stable meniscus, characterized by a cone with a half angle of approximately 49.2 degrees. This so-called Taylor cone, characteristic of conventional electrospray droplet formation, anchors a thin jet at its tip. This thin jet extends briefly toward the electrode as it breaks into a series of uniformly charged droplets. These droplets propagate toward the electrode via the action of the field before finally breaking off. In other embodiments, an increase in the electrostatic drag force relative to the surface tension creates a pulsating meniscus, which generates droplets by dripping. During each cycle of pulsation, the meniscus extends toward the electrode in the form of a jet, which eventually breaks off from the base of the meniscus. As the meniscus contracts in preparation for the next cycle, the detached jet either forms a single droplet or breaks off into a collection of droplets. In yet other embodiments, there are a variety of different modes of electrospray.The exact mode in which electrically charged droplets are generated generally depends at least on the conductivity, polarizability, viscosity, and surface tension coefficient of the electrosprayed liquid, and the geometry of the electric field (M. Cloupeau and B. Prunet-Foch, Electrostatic spraying of liquids: Main functioning modes, J. Electrostatics, 25, 165-184, 1990; M. Cloupeau and B. Prunet-Foch, J. Aerosol Sci., vol. 25, no. 6, pp. 1021-1036, 1994).

[0117] In some embodiments, electrospray technology is utilized by supplementing a nebulizer, microfluidic device, or some other primary droplet-forming device with an electric field to form electrospray droplets. In some embodiments, this can reduce the amount of energy that this primary device contributes to the formation of each droplet. In the case of an ultrasonic nebulizer, for example, the presence of an electric field can potentially reduce the minimum power required by the ultrasonic generator to generate droplets. This can be advantageous in that it reduces selectivity effects, such as cavitation, that can affect the integrity of certain therapeutic or diagnostic agents in the first liquid (S. Vonhoff, The Influence of Atomization Conditions on Protein Secondary and Tertiary Structure During Microparticle Formation by Spray-Freeze-Drying, PhD Thesis, University of Erlangen-Nuremberg, 2010). In certain embodiments, the electric field can also reduce the average droplet size and / or narrow the droplet dispersity compared to what can be achieved in the absence of an electric field.

[0118] Drying of the droplets, i.e., removal of the first liquid to produce dried particles, can be accomplished via any of several methods known in the art. These include, but are not limited to, warm gas evaporation, freeze-drying, critical point drying, emulsion solvent evaporation, emulsion solvent diffusion (US8013022; US8512754), and combinations thereof. In some embodiments, the primary drying step is followed by a secondary drying step, such as freeze-drying or vacuum drying, intended to further reduce the residual amount of the first liquid within the particles. In some embodiments, the residual amount of the first liquid in the particles after primary or secondary drying is 0-10% by weight, e.g., 0-5% by weight, or 0-3% by weight, or 0-1% by weight.

[0119] In some embodiments, the effect of the electric field is such that a reversible or irreversible charge is induced on the therapeutic or diagnostic agent in the droplets of the first liquid, i.e., the agent is ionized (Anal. Chem., 2005, 77, 5370). This effect can be tuned by controlling certain electrospray parameters, such as the size of the droplets and their composition. In some embodiments, the inclusion of certain additives, e.g., amino acids, stabilizes the charged therapeutic or diagnostic agent. In other embodiments, the additives preferentially carry a charge, making the therapeutic or diagnostic agent less susceptible to ionization. In some embodiments, the charge on the droplets is reduced or even completely neutralized during drying and / or during contact between the particles and the electrode. In other embodiments, some of this charge is intentionally preserved by preventing direct contact between the particles and the electrode.

[0120] In some embodiments, the effect of the electric field is such that free charges and / or polar molecules preferentially migrate to the surface of the droplets of the first liquid due to the Coulomb effect. The former phenomenon, localization of free charges at the interface between the first liquid and the dielectric medium where the droplets are formed, creates a layer of surface charge. In some embodiments, such an effect is exploited to influence the structure and / or surface properties of the droplets and / or particles. In some embodiments, for example, the orientation of the first liquid near the surface of the droplets facilitates faster removal of the first liquid, possibly at lower temperatures. The ability to achieve low residual moisture content in primary drying may also be improved. This may be particularly beneficial when drying with a warm gas stream, where the temperature of the gas stream may be correlated with degradation of the therapeutic or diagnostic agent.

[0121] In some embodiments, the effect of the electric field is such that free charges and / or polar molecules preferentially migrate to the surface of the droplet of the first liquid due to the Coulomb effect, causing the therapeutic or diagnostic agent to crystallize. Nucleation of the agent can be controlled to preferentially obtain a desired polymorph (A. Ziabicki, L. Jarecki, Macromolecular Symposia, 1996, 104, 65-87).

[0122] In some embodiments, core-shell particles are produced by coaxial electrospray (FIG. 4). In this case, a second liquid containing dissolved encapsulant or shell material is provided along with the first liquid during electrospraying. This is typically accomplished by flowing the second liquid through an annular tube that is aligned coaxially with the tube through which the first liquid flows. At the end of this coaxial tube arrangement, an electrospray is formed, in which droplets comprise a core of the first liquid and a shell of the second liquid. Drying of the droplets can proceed via any of the usual routes. In other embodiments, core-shell particles are formed from electrospray of only the first liquid by exploiting the tendency of certain polar molecules and free charges to be located at the surface of the droplets. In certain cases, this results in localization of therapeutic or diagnostic agents to the core or surface of the droplets, where they can be preserved during drying. In some embodiments, this involves deterministic layering of various agents (e.g., therapeutic agents, diagnostic agents, additives) throughout the thickness of the particle. In certain embodiments, non-therapeutic components such as salts (e.g., NaCl) or sugars (e.g., sucrose) are preferentially driven to the surface by the electric field and crystallize or otherwise form a thin shell around the particle. This shell may have a protective effect or provide a means of control over pharmacokinetics. In other embodiments, a portion of the drug may be localized to the particle surface without necessarily forming a uniform or continuous shell (Figures 22, 23).

[0123] Treatment drugs

[0124] Exemplary therapeutic or diagnostic agents include nucleic acids, oligonucleotides, antibodies, amino acids, peptides, proteins, cells, bacteria, gene therapy drugs, genome-engineered therapeutic drugs, epigenome-engineered therapeutic drugs, carbohydrates, chemical drugs, contrast agents, magnetic particles, polymer beads, metal nanoparticles, metal microparticles, quantum dots, antioxidants, antibiotics, hormones, nucleoproteins, polysaccharides, glycoproteins, lipoproteins, steroids, analgesics, local anesthetics, anti-inflammatory agents, antimicrobial agents, chemotherapeutic agents, exosomes, outer membrane vesicles, vaccines, viruses, bacteriophages, adjuvants, vitamins, minerals, organelles, and combinations thereof (Table 1). Therapeutic and diagnostic agents may have a molecular weight of 20 to 200 kDa, e.g., 40 to 150 kDa. The concentration of the therapeutic or diagnostic agent in the droplet is typically at least 1 mg / mL, e.g., at least 5 mg / mL, at least 10 mg / mL, at least 50 mg / mL, at least 100 mg / mL, or at least 500 mg / mL. The first therapeutic or diagnostic agent in the droplets can have an activity of 0.5 to 1.0 activity / unit, 0.75 to 1.0 activity / unit, 0.9 to 1.0 activity / unit, 0.95 to 1.0 activity / unit, or 0.99 to 1.0 activity / unit, where the activity is measured relative to the same therapeutic or diagnostic agent before it is electrosprayed.

[0125] [Table 2]

[0126] In some embodiments of any of the foregoing methods, the therapeutic and diagnostic agent is an antibody. In some embodiments, the antibody is selected from the group consisting of 3F8, abagovomab, abciximab, abituzumab, abrilumab, acritumomab, actoxumab, adalimumab, adalimumab-atto, adecatumumab, Ado-trastuzumab emtansine, aducanumab, afacevicumab, afelimomab, afutuzumab, alacizumab pegol, ALD518, alemtuzumab, alirocumab, altumomab pentetate, amatuximab, anatumomab mafenatox, anetumab ravtansine, anifrolumab, and anrukinz. Mab, apolizumab, arcitumomab, ascribacumab, azelizumab, atezolizumab, atinumab, atlizumab, atollimumab, avelumab, bapineuzumab, basiliximab, bavituximab, bectumomab, begelomab, belimumab, benralizumab, bertilimumab, besilesomab, bevacizumab, bezlotoxumab, biciromab, bimagrumab, bimekizumab, bivatuzumab mertansine, bleselumab, blinatumomab, brontuzumab, brosozumab, bococizumab, brazikumab, brentuximab Buvedotin, briakinumab, brodalumab, brolucizumab, brontixutuzumab, burosumab, cabilalizumab, canakinumab, cantuzumab mertansine, cantuzumab ravtansine, caplacizumab, capromab pendetide, carlumab, carotuximab, catumaxomab, cBR96-doxorubicin immunoconjugate, cedelizumab, sergituzumab amnaleukin, certolizumab pegol, cetuximab, sitatuzumab bogatox, cixutumumab, clazakizumab, clenoliximab, clivatuzumab tetraxetan , codrituzumab, coltuximab ravtansine, conatumumab, concizumab, crenezumab, clotedumab, CR6261, dacetuzumab, daclizumab, darotuzumab, dapirolizumab pegol, daratumumab, dectrecumab, demcizumab, denintuzumab mafodotin, denosumab, depatuxizumab mafodotin, delrotuximab biotin, detumomab, dinutuximab, zilidabumab, domagurozumab, dorlimomab alitoxin, drozitumab, durigotumab, dupilumab, durvalumab, dusigitumab,Eclomeximab, eculizumab, edovacomab, edrecolomab, efalizumab, efungumab, eldelumab, elgemtumab, elotuzumab, ersilimomab, emactuzumab, emibetuzumab, emicizumab, enavatuzumab, enfortumab vedotin, enlimomab pegol, enoblitzumab, enokizumab, enoticumab, ensituximab, epitumomab cituxetan, epratuzumab, erenumab, erlizumab, ertumaxomab, etaracizumab, etrolizumab, evinacumab, evolocumab, exibirmab, fanoresoma , faralimomab, farletuzumab, fasinumab, felvizumab, fezakinumab, fibatuzumab, ficlatuzumab, figitumumab, filibumab, framvotumab, fretikumab, fontolizumab, foralumab, foravirumab, fresolimumab, furanumab, futuximab, galcanezumab, galiximab, ganitumab, gantenerumab, gavilimomab, gemtuzumab ozogamicin, gevokizumab, girentuximab, glenbatumumab vedotin, golimumab, gomiliximab, guselkumab, ibalizumab, ibritumomab Uxetan, icrucumab, idarucizumab, igovomab, IMAB362, imalumab, imciromab, imgatuzumab, inlacumab, indatuximab ravtansine, indusatumab vedotin, inebilizumab, infliximab, infliximab-dyyb, intetumumab, inolimomab, inotuzumab ozogamicin, ipilimumab, iratumumab, isatuximab, itolizumab, ixekizumab, keliximab, labetuzumab, lambrolizumab, lampalizumab, lanadelumab, landgrozumab, laprituximab emtansine , lebrikizumab, remaresomab, lendalizumab, lendilumab, lerdelimumab, lexatumumab, ribivirumab, rifastuzumab vedotin, ligelizumab, rilotomab satetraxetan, lintuzumab, lirilumab, roderucizumab, loxivetumab, lorvotuzumab mertansine, lucatumumab, lurizumab pegol, lumiliximab, lumletuzumab, mapatumumab, marjetuximab, maslimomab, mavrilimumab, matuzumab, mepolizumab, metelimumab, milatuzumab, minletumomab, mirvetuximab soravtansine, mitsumomab,Mogamulizumab, monalizumab, morolimu- mab, motavizumab, moxetumomab pasudotox, muromonab-CD3, nacolomab butafenatox, namilumab, naptumomab estafenatox, naratuximab emtansine, narutumab, natalizumab, nabicixizumab, nabibu- mab, nebacumab, necitumumab, nemolizumab, nerelimomab, nesbacumab, nimotuzumab, nivolumab, nofetumomab merpentane, obilutoxaximab, obinutuzumab, occaratuzumab, ocrelizumab, odulimomab, ofatumumab, olaratumab, Olokizumab, omalizumab, onartuzumab, ontuxizumab, opicinumab, oportuzumab monatox, oregovomab, olticumab, otelixizumab, otreltuzumab, oxelumab, ozoralizumab, pasivaximab, palivizumab, pamrevlumab, panitumumab, pancomab, panobacumab, palsatuzumab, pascolizumab, pasotuxizumab, pateclizumab, patritumab, pembrolizumab, pemtumomab, perakizumab, pertuzumab, pexelizumab, pidilizumab, pinatuzumab vedotin, pintumomab , placumab, prosalizumab, pogalizumab, polatuzumab vedotin, ponezumab, prezalizumab, priliximab, plitoxaximab, pritumumab, PRO140, kilimumab, racotumomab, radletumab, rafivirumab, ralpancizumab, ramucirumab, ranibizumab, raxibacumab, refanezumab, regavirumab, reslizumab, rilotumumab, rinukumab, risankizumab, rituximab, rivavazumab pegol, lobatumumab, loredumab, romosozumab, rontalizumab, rovalpituzumab tesirin, rovelizumab, ru Prizumab, sacituzumab govitecan, samalizumab, sapelizumab, sarilumab, satumomab pendetide, secukinumab, seribantumab, cetoxaximab, sevilumab, sibrotuzumab, SGN-CD19A, SGN-CD33A, sifalimumab, siltuximab, simtuzumab, siplizumab, sirukumab, sofituzumab vedotin, solanezumab, solitomab, sonepcizumab, sontuzumab, stamulumab, sulesomab, subizumab, tabalumab, tacatuzumab tetraxetan, tadocizumab, talizumab, tamtubetumab, tanezumab,Taplitumomab paptox, talexuzumab, tefibazumab, terimomab alitox, tenatumomab, teneliximab, teplizumab, teprotumumab, tesidolumab, tetulomab, tezepelumab, TGN1412, ticilimumab, tildrakizumab, tigatuzumab, timolumab, tisotumab vedotin, TNX-650, tocilizumab, toralizumab, tosatoxumab, tositumomab, tobetumab, tralokinumab, trastuzumab, trastuzumab emtansine, tregalizumab, tremelimumab, trevoglumab, tucotuzumab-cel Molleukin, tuvilumab, ublituximab, urocupulumab, urelumab, urtoxazumab, ustekinumab, utomilumab, vadastuximab butarilin, bundletuzumab vedotin, vanticutumab, vanucizumab, bapaliximab, varlilumab, batelizumab, vedolizumab, veltuzumab, bepalimomab, besencumab, bisilizumab, bovalilizumab, volociximab, borsetuzumab mafodotin, votumumab, xentuzumab, zalutumumab, zanolimumab, zatuximab, diralimumab, or zolimomab alitox.

[0127] In some embodiments, the therapeutic agent is an immunotherapeutic agent. In some embodiments, the immunotherapeutic agent is a PD-1 inhibitor such as a PD-1 antibody, a PD-L1 inhibitor such as a PD-L1 antibody, a CTLA-4 inhibitor such as a CTLA-4 antibody, a CSF-1R inhibitor, an IDO inhibitor, an A1 adenosine inhibitor, an A2A adenosine inhibitor, an A2B adenosine inhibitor, an A3A adenosine inhibitor, an arginase inhibitor, or an HDAC inhibitor. In some embodiments, the immunotherapeutic agent is a PD-1 inhibitor (e.g., nivolumab, pembrolizumab, pidilizumab, BMS936559, and MPDL328OA). In some embodiments, the immunotherapeutic agent is a PD-L1 inhibitor (e.g., atezolizumab and MEDI4736). In some embodiments, the immunotherapeutic agent is a CTLA-4 inhibitor (e.g., ipilimumab). In some embodiments, the immunotherapeutic agent is a CSF-1R inhibitor (e.g., pexidartinib and AZD6495). In some embodiments, the immunotherapeutic agent is an IDO inhibitor (e.g., norharmane, rosmarinic acid, and alpha-methyl-tryptophan). In some embodiments, the immunotherapeutic agent is an A1 adenosine inhibitor (e.g., 8-cyclopentyl-1,3-dimethylxanthine, 8-cyclopentyl-1,3-dipropylxanthine, 8-phenyl-1,3-dipropylxanthine, bamifylline, BG-9719, BG-9928, FK-453, FK-838, rolofylline, or N-0861). In some embodiments, the immunotherapeutic agent is an A2A adenosine inhibitor (e.g., ATL-4444, istradefylline, MSX-3, preladenant, SCH-58261, SCH-412,348, SCH-442,416, ST-1535, VER-6623, VER-6947, VER-7835, viadenant, or ZM-241,385). In some embodiments, the immunotherapeutic agent is an A2B adenosine inhibitor (e.g., ATL-801, CVT-6883, MRS-1706, MRS-1754, OSIP-339,391, PSB-603, PSB-0788, or PSB-1115).In some embodiments, the immunotherapeutic agent is an A3A adenosine inhibitor (e.g., KF-26777, MRS-545, MRS-1191, MRS-1220, MRS-1334, MRS-1523, MRS-3777, MRE-3005-F20, MRE-3008-F20, PSB-11, OT-7999, VUF-5574, and SSR161421). In some embodiments, the immunotherapeutic agent is an arginase inhibitor (e.g., an arginase antibody, (2s)-(+)-amino-5-iodoacetamidopentanoic acid, NG-hydroxy-L-arginine, (2S)-(+)-amino-6-iodoacetamidohexanoic acid, or (R)-2-amino-6-borono-2-(2-(piperidin-1-yl)ethyl)hexanoic acid). In some embodiments, the immunotherapeutic agent is an HDAC inhibitor (eg, valproic acid, SAHA, or romidepsin).

[0128] In some embodiments, the therapeutic agent can be ledipasvir / sofosbuvir, insulin glargine, lenalidomide, pneumococcal 13-valent conjugate vaccine, fluticasone / salmeterol, elvitegravir / cobicistat / emtricitabine / tenofovir alafenamide, emtricitabine, rilpivirine and tenofovir alafenamide, emtricitabine / tenofovir alafenamide, grazoprevir / elbasvir, coagulation factor VIIa recombinant, epoetin alfa, aflibercept, or etanercept.

[0129] In some embodiments, the therapeutic or diagnostic agent is abatacept, abobotulinumtoxinA, agalsidase beta, albiglutide, aldesleukin, alglucosidase alfa, alteplase (cathfloactivase), anakinra, asfotase alfa, asparaginase, asparaginase from Erwinia chrysanthemi, becaplermin, belatacept, collagenase, clostridium histolyticum-derived collagenase, darbepoetin alfa, denileukin diftitox, dornase alfa, dulaglutide, ecallantide, elosulfase alfa, etanercept-szzs, filgrastim, filgrastim-sndz, galsulfase, glucarpidase, idursulfase, incobotulinumtoxin A, interferon alfa-2b, interferon alfa-n3, interferon beta-1a, interferon beta-1b, interferon gamma-1b, laronidase, methoxypolyethylene glycol-epoetin beta, metreleptin, ocriplasmin, onabotulinumtoxinA, oprelvekin, palifermin, parathyroid hormone, pegaspargase, pegfilgrastim, peginterferon alfa-2a, peginterferon alfa-2a packaged with ribavirin, peginterferon alfa-2b, peginterferon beta-1a, pegloticase, rasburicase, reteplase, rilonacept, rimabotulinumtoxinB, romiplostim, sargramostim, sebelipase alfa, Tbo-filgrastim, tenecteplase, or Ziv-aflibercept.

[0130] In some embodiments, the diagnostic agent is tuberculin purified protein derivative, thyrotropin alpha, secretin, soluble transferrin receptor, troponin, B-type natriuretic peptide, iobenguane I 123, florbetapir F 18, perflutren, gadoterate meglumine, florbetaben F 18, flutemetamol F 18, gadoterate meglumine, isosulfan blue, regadenoson, technetium Tc 99m tilmanocept, florbetaben F 18, perflutren, regadenoson, or flutemetamol F 18.

[0131] formulation

[0132] The electrosprayed particles can be suspended in a non-aqueous or aqueous liquid or gel (or a mixture thereof) to form a suspension formulation. The non-aqueous liquid can be an organic solvent or an ionic liquid, or some combination thereof. In some embodiments, the organic solvent is selected from the group consisting of benzyl alcohol, benzyl benzoate, coconut oil, cottonseed oil, fish oil, grapeseed oil, hazelnut oil, hydrogenated vegetable oil, olive oil, palm kernel oil, peanut oil, peppermint oil, safflower oil, sesame oil, soybean oil, sunflower oil, walnut oil, acetone, ethyl acetate, ethyl lactate, dimethylacetamide, dimethylisosorbide, dimethyl sulfoxide, glycofurol, diglyme, methyl tert-butyl ether, N-methylpyrrolidone, perfluorodecalin, polyethylene glycol, 2-pyrrolidone, tetrahydrofurfuryl alcohol, and the like. ethanol, triglycerides, fractionated triglycerides of C8 and C10 vegetable fatty acids (e.g., MIGLYOL® 810 and MIGLOYL® 812N), propylene glycol diesters of saturated C8 and C10 vegetable fatty acids (e.g., MIGLYOL® 840), ethyl oleate, ethyl caprate, dibutyl adipate, fatty acid esters, hexanoic acid, octanoic acid, triacetin, diethyl glycol monoether, gamma-butyrolactone, eugenol, clove bud oil, citral, limonene, and any combination thereof. In some embodiments, the ionic liquid is pyridinium, pyridazinium, pyrimidinium, pyrazinium, imidazolium, pyrazolium, thiazolium, oxazolium, triazolium, ammonium, sulfonium, halide, sulfate, sulfonate, carbonate, phosphate, bicarbonate, nitrate, acetate, PF6 - , BF4 -, triflate, nonaflate, bis(trifyl)amide, trifluoroacetate, heptafluorobutanoate, haloaluminate, or any combination thereof. Aqueous liquids for suspension include water, 0.9% saline, lactated Ringer's solution, dextrose 5%, or buffer solutions. Buffer solutions may include, for example, acetate buffer, histidine buffer, succinate buffer, HEPES buffer, Tris buffer, carbonate buffer, citrate buffer, phosphate buffer, glycine buffer, barbital buffer, and cacodylate buffer. The suspension medium may further include other components, such as carbohydrates, pH adjusters, salts, chelating agents, minerals, polymers, surfactants, protein stabilizers, emulsifiers, preservatives, amino acids, antioxidants, proteins, organic solvents, or nutrient media. In some embodiments, each of the other components independently constitutes 0.0001 to 99% (w / v) of the vehicle, e.g., 0.0001 to 90% (w / v), 0.0001 to 50% (w / v), 0.0001 to 10% (w / v), 0.0001 to 1% (w / v), or 0.0001 to 0.1% (w / v). One of skill in the art would be able to determine the appropriate amounts of the other components in the vehicle. The carbohydrate may be dextran, trehalose, sucrose, agarose, mannitol, lactose, sorbitol, or maltose. The pH adjuster may be, for example, acetate, citrate, glutamate, glycinate, histidine, lactate, maleate, phosphate, succinate, tartrate, bicarbonate, aluminum hydroxide, phosphoric acid, hydrochloric acid, DL-lactic acid / glycolic acid, phosphorylethanolamine, tromethamine, imidazole, glycylglycine, or monosodium glutamate. Exemplary salts include sodium chloride, calcium chloride, potassium chloride, sodium hydroxide, stannous chloride, magnesium sulfate, sodium glucoheptonate, sodium pertechnetate, or guanidine hydrochloride. The chelating agent may be, for example, disodium edetate. The mineral may be, for example, calcium, zinc, or titanium dioxide.Examples of polymers include propylene glycol, glucose star polymers, silicone polymers, polydimethylsiloxane, polyethylene glycol, carboxymethylcellulose, poly(glycolic acid), poly(lactic acid-co-glycolic acid), or polylactic acid. Surfactants can be, for example, polysorbate, magnesium stearate, sodium dodecyl sulfate, Triton N-101, glycerin, or polyoxyethylated castor oil. Protein stabilizers include acetyltryptophan salts, caprylate salts, or N-acetyltryptophan. Emulsifiers can be, for example, polysorbate 80, polysorbate 20, sorbitan monooleate, ethanolamine, polyoxyl 35 castor oil, polyoxyl 40 hydrogenated castor oil, carbomer 1342, corn oil mono-di-triglyceride, polyoxyethylated oleic acid glyceride, or poloxamer. Exemplary preservatives include phenol, m-cresol, benzyl alcohol, 2-phenyloxyethanol, chlorobutanol, neomycin, benzethonium chloride, glutaraldehyde, or beta-propiolactone. The amino acid may be alanine, aspartic acid, cysteine, isoleucine, glutamic acid, leucine, methionine, phenylalanine, pyrrolidine, serine, selenocysteine, threonine, tryptophan, tyrosine, valine, asparagine, L-arginine, histidine, glycine, or glutamine, such as asparagine, L-arginine, histidine, glycine, or glutamine. The antioxidant may be glutathione, ascorbic acid, cysteine, or tocopherol. The protein may be protamine, protamine sulfate, or gelatin.

[0133] For aqueous suspension formulations, high-concentration trehalose solutions can stabilize particles in suspension and prevent premature dissolution. The sugar acts as a "crowder" molecule, i.e., a crowding agent that occupies a large volume and, in some embodiments, increases short-range protein-protein attractive interactions. This stabilizing effect has also been described for other crowding agents in water, such as polymers, e.g., PEG300, and organic molecules, e.g., N-methyl-2-pyrrolidone (Miller et al. J. Pharm. Sci., 2012, 101, 3763-3778).

[0134] The viscosity of the suspension can range from 0.27 to 200 cP, e.g., 0.27 to 50 cP, 1 to 30 cP, or 20 to 50 cP. In some embodiments, the viscosity of the suspension ranges from 0.27 to 200 cP, e.g., 0.27 to 100 cP, 0.27 to 50 cP, 0.27 to 30 cP, 1 to 20 cP, or 1 to 15 cP. In certain embodiments, the viscosity is measured at a shear rate in the Newtonian region. In other embodiments, the viscosity is measured at a shear rate of 100 s -1 More than, for example, 1000s -1 or 1000s -1 The suspension may comprise 5-90% by volume of particles, such as, for example, 20-90%, 40-80%, 50-60%, or 70-90% by volume of particles. The suspension may have a concentration of the first therapeutic or diagnostic agent of 0.0001-1000 mg / mL, e.g., 100-900, 150-800, or 200-700 mg / mL.

[0135] In some embodiments of the invention described herein, high concentrations of therapeutic or diagnostic agents in particles and high concentrations of particles in non-aqueous or aqueous liquids are possible, the latter of which can be achieved by mixing particles of various sizes.

[0136] In some embodiments, one or more therapeutic or diagnostic agents can be present in the particles and outside the particles, i.e., in a non-aqueous or aqueous liquid. The therapeutic or diagnostic agent contained in the non-aqueous or aqueous liquid can be the same as or different from that used in the particles. The one or more therapeutic or diagnostic agents can reduce pain or inflammation during administration. The concentration of the therapeutic agent in the pharmaceutical composition outside the particles can range, for example, from 0.0001 to 1000 mg / mL.

[0137] In some embodiments, the particles are nebulized and collected for use in a needleless injector or for use in an inhalation or other nasal delivery system. In some embodiments, the particles are stored as a dry powder. In some embodiments, the dry powder is reconstituted immediately prior to administration of the formulation, in which the therapeutic or diagnostic agent is dissolved in an aqueous or non-aqueous solution. Such a paradigm may be beneficial in some cases to avoid the relative instability or degradation of certain therapeutic or diagnostic agents that are stored in aqueous or non-aqueous solution form rather than in dry powder form.

[0138] Needle-free injection systems may include liquids, suspensions, powders, or projectiles. Powders are suitable for long-term storage and can be injected at home using such systems without prior reformulation and preparation. Powders require an injection chamber filled with solid drug and a nozzle for projecting particles into the skin. Briefly, in some embodiments, particles exit the nozzle with a gas stream and impact the skin surface, creating small perforations or holes where the particles are deposited. After penetrating the stratum corneum, they are completely distributed within the stratum corneum and the biological epidermis. Particles for needle-free injection have a density of approximately 1 g / cm. 3 The particles may have a density of 100 μm or more and an average diameter of 20 μm or more.

[0139] In some embodiments, the particles are administered via a dual-chamber syringe device, such as the LyoTwist. The particles are present as a dry powder in one chamber of the device, while the second chamber is occupied by an aqueous or non-aqueous carrier liquid. The components of the two chambers are mixed immediately prior to administration.

[0140] How to use

[0141] The pharmaceutical compositions of the present invention, including suspensions or dry forms, can be administered at appropriate dosages, which can be adjusted as needed depending on the clinical response. The compositions can also be used cosmetically. The dosage of the pharmaceutical composition can vary depending on many factors, such as the pharmacokinetics of the therapeutic or diagnostic agent; the mode of administration; the recipient's age, health, and weight; the nature and severity of symptoms; the frequency of treatment and, if present, the type of concomitant treatment; and the clearance rate of the therapeutic or diagnostic agent in the treated animal. Those skilled in the art can determine the appropriate dosage based on the above factors. Administration may be daily, weekly, every two weeks, every three weeks, monthly, or at any other suitable interval. Generally, satisfactory results can be obtained when the therapeutic or diagnostic agent is administered to humans at a dosage of, for example, 0.01 mg / kg to 70 mg / kg (measured as solid form). In some embodiments, the dosage can range from 0.01 mg / kg to 1 mg / kg. Dose ranges include, for example, between 30 mg and 5000 mg. In some embodiments, at least 30, 100, 500, 1000, 2000, or 5000 mg of the compound is administered. Preferred dose ranges include, for example, 1-30 mg / kg, such as 1-10 mg / kg.

[0142] The volume delivered will depend on the indication and the route of delivery. In one embodiment, a pharmaceutical composition having a viscosity of less than 50 cP, e.g., less than 30 cP, in a volume greater than 0.5 mL, e.g., greater than 2 mL, is administered, e.g., injected, into the skin of an animal. In some embodiments, a pharmaceutical composition having a viscosity of less than 50 cP, e.g., less than 30 cP, in a volume greater than 1.5 mL, e.g., greater than 5 mL, is administered, e.g., injected, into the skin of an animal.

[0143] The pharmaceutical composition can be administered by any suitable method, for example, into the auricle, buccal, conjunctival, skin, teeth, electroosmotic, intracervical, intrasinus, intratracheal, enteral, epidural, extraamniotic, extracorporeal, infiltrative, interstitial, intraperitoneal, intraamniotic, intra-arterial, intra-articular, intra-biliary, intra-bronchial, intra-vesical, intracardiac, intracartilaginous, intrasacral, intra-cavity, intra-cavity, intracerebral, intra-cisternal, intra-corneal, intra-coronary, intra-coronary, intra-cavernous, intradermal, intra-discal, intra-ductal, intra-duodenal, intra-dural, intra-epidermal, intra-esophageal, intra-gastric, intra-gingival, intra-ileal, intra-lesional, intra-luminal, intra-lymphatic, intra-medullary, intra-meningeal, intra-muscular, intra-ocular, intra-ovarian, intra-pericardial, intra-peritoneal, intra-pleural, intra-pleural. The pharmaceutical composition may be administered intraprostatically, intrapulmonary, intranasal or periorbital, intraspinal, intrasynovial, intratendinous, intratesticular, intrathecal, intrathoracic, intrarenal tubular, intratumor, intratympanic, intrauterine, intravascular, intravenous, intravenous bolus, intravenous infusion, intraventricular, intravesical, intravitreal, iontophoretic, lavage, laryngeal, nasal, nasogastric, occlusive dressing, ocular, oral, oropharyngeal, parenteral, transdermal, periarticular, epidural, perineural, periodontal, rectal, inhalation, retrobulbar, soft tissue, subarachnoid, subconjunctival, subcutaneous, sublingual, submucosal, topical, transdermal, transmucosal, transplacental, transtracheal, transtympanic, ureteral, urethral, ​​or vaginal. The pharmaceutical composition may then be appropriately formulated. [Example]

[0144] example

[0145] The method disclosed herein was utilized in separate cases to produce particles containing bovine serum albumin (BSA), whole human IgG, and one of three separate monoclonal antibodies. Various analytical techniques were applied to evaluate the physical properties of the particles themselves and the structural and functional properties of the proteins they contain. Scanning electron microscopy and associated image analysis were used to study particle morphology and size distribution, respectively. Highly spherical, smooth particles were achieved for certain additive profiles, while easy control of average size was possible over a wide range with low dispersity. In certain cases, the particle surface was decorated with additives. Density and water content measurements showed that the particles approached crystalline packing efficiency and retained very low levels of residual moisture, especially when secondary drying was used. The functional properties of the proteins were also preserved, as evidenced by ELISA and binding assays performed on the reconstituted particles. This was supported by size-exclusion HPLC analysis, which showed that the process had minimal, or even reversible, effects on the degree of protein-protein association. Finally, suspensions of particles in various carrier media showed that ultra-high loadings, eg, >300 mg / mL, were possible, with apparent viscosities of the particle-vehicle complexes below 20 cP.

[0146] Materials and Methods

[0147] Bovine serum albumin and human IgG were obtained as lyophilized powders from Sigma-Aldrich (A2934, >98%) and Innovative Research (IRHUGGF-LY, >97%), respectively. Biosimilar versions of three commercially available monoclonal antibody products (mAb1, mAb2, mAb3) were obtained in aqueous solution. Formulations conformed to the corresponding FDA label. Concentration columns were obtained from Millipore Sigma (Amicon® Ultra 15 mL filters for protein purification and concentration with a 3 kDa cutoff) and used as needed to achieve the desired protein concentration for the electrospray feed solution. All additives and carrier media, including benzyl benzoate, were purchased from Sigma-Aldrich and used as received.

[0148] Electrospray particle formation

[0149] Unless otherwise noted, the particle samples disclosed herein were prepared using a custom-designed conventional electrospray device. The device included a 30-gauge blunt disposable syringe needle (89134-196, VWR International) attached to a Harvard Apparatus Model 33 dual-channel syringe pump. The needle was charged by a Matsuda EQ-30P1-LCtG high-voltage DC power supply. For selected samples, the needle was replaced by a Sono-Tek 120 kHz ultrasonic atomizer nozzle driven by 4.5 W of power.

[0150] Freeze drying

[0151] Freeze-dried samples, i.e., particles of samples marked as having undergone secondary drying, were placed in microcentrifuge tubes and flash-frozen by immersion in liquid nitrogen for approximately 10 minutes. The samples were then loosely covered and placed in a Labconoco Freezone freeze dryer at approximately 0.035 Torr pressure for approximately 48 hours.

[0152] Scanning electron microscopy

[0153] Electron micrographs were collected on selected samples using a Hitachi TM3030Plus tabletop microscope. Samples were mounted on conductive tape and tested in a low vacuum antistatic environment, eliminating the need for sample preparation.

[0154] optical microscopy

[0155] Selected samples were prepared for imaging by vortexing 5 mg of particles with 20 microliters of benzyl benzoate, then pipetting the samples onto glass slides and imaging them using a Celena microscope from Logos Biosystems.

[0156] Image analysis

[0157] Selected microscopy images were chosen for further analysis based on (i) minimal particle overlap, (ii) good contrast between particles and the background, and (iii) a resolution that provided a particle occupancy of at least 10 pixels. This allowed for easy particle identification and reduced resolution-based errors. A binarization threshold was applied to separate particles from the background, and a watershed segmentation algorithm was applied to ensure that individual particles were measured separately. The ImageJ tool "Analyze Particles" was then applied to the binary images with the following parameters: circularity between 0.5 and 1.0; size between 5 and infinity square micrometers; exclusion on edges; and hole filling. The outlines of identified particles were overlaid on the original image. Misidentified particles, such as clusters identified as single particles or particles whose outlines did not match any particle, were then discarded. Missing particles were measured by manually tracing particle outlines and using ImageJ's measurement tools.

[0158] Density analysis

[0159] The skeletal density of electrosprayed particles from selected samples was determined by examining approximately 0.1 g of powder using an AccuPyc II 1340 gas displacement pycnometry system.

[0160] water content analysis

[0161] Residual moisture in electrosprayed particles from selected samples was determined by placing approximately 0.1 g of powder in a vacuum oven equipped with a Karl Fischer titrator and heating the sample.

[0162] Salt content analysis

[0163] The salt content of electrosprayed particles from selected samples was determined by elemental analysis of chlorine, which was performed by flask combustion followed by ion chromatography.

[0164] Zeta potential analysis

[0165] Zeta potential analysis was performed on selected samples using a Malvern Instruments Zetasizer Nano ZS. Solutions were prepared at 8 mg / mL in DI water and placed in a Malvern Instruments INC DTS1070 folded capillary cell.

[0166] ELISA assay

[0167] An ELISA assay was used on select samples to detect human antibodies in a denaturing-sensitive format. Human IgG was first plated in PBS for 1 hour, followed by three 4-minute washes with wash buffer (PBS + 0.05% Tween 20), followed by blocking with 2% BSA (Sigma) in wash buffer for 45 minutes, followed by incubation with diluted (20 μg / ml) Protein A-HRP (Abcam) for 45 minutes, three 3-minute washes with wash buffer, followed by incubation with TMB (Abcam) for 10 minutes, and finally quenching the reaction with stop solution (Abcam). Colorimetric readings were performed on a Thermo Multiskan Spectrum.

[0168] Monoclonal antibody binding assay

[0169] Monoclonal antibodies from selected samples were evaluated for cell binding ability using cells expressing the appropriate cell surface receptor. Cells were incubated with each concentration of monoclonal antibody for 30 minutes at 4°C, then spun down at 2000 rpm and washed three times with PBS. Cells were then incubated with a secondary goat anti-human Fab antibody fluorescently labeled with PE for 30 minutes at 4°C. Cells were then spun down at 2000 rpm and washed three times with PBS. Cells were then resuspended and analyzed using an Attune flow cytometer (Invitrogen).

[0170] Size exclusion chromatography

[0171] Quantification of size variants in selected samples was determined by size exclusion chromatography. The analysis utilized an Advanced BioSEC column, 7.8 mm ID x 30 cm, 3 μm (Agilent), run on an HPLC system (1100, Agilent). The mobile phase was 0.2 M potassium phosphate and 0.25 M potassium chloride at pH 6.0. Chromatography was performed isocratically for 15 min at a flow rate of 1.0 mL / min. The column temperature was maintained at ambient temperature of 25°C, and the eluent absorbance was monitored at 280 nm. Each monoclonal antibody was diluted to 1 mg / mL in its respective formulation buffer. The injection volume was 20 μL.

[0172] Suspension preparation

[0173] For selected samples, particles were weighed into a 2 mL Eppendorf microcentrifuge tube. Based on the measured powder density, an appropriate amount of suspension vehicle was added to prepare a suspension of the desired concentration. The sample was then vortexed for 30 seconds.

[0174] Viscosity measurement

[0175] The viscosity of solutions and suspensions was measured using an AR-G2 rheometer (TA Instruments) with a cone and plate geometry (20 mm / 2°). Samples were measured every 30 seconds for 5 minutes at a shear rate of 1000 / s. In certain cases, the intrinsic viscosity coefficient [η] was calculated based on the measurements. This was an indicator of the degree to which particles contributed to the viscosity of the suspension. It was calculated from the Krieger-Dougherty equation:

number

[0176] Results and Discussion

[0177] Particle size, shape, and chemical composition

[0178] A 4 mL solution of BSA was prepared in deionized water at a concentration of approximately 80 mg / mL. This solution was electrosprayed at a flow rate of 0.4 mL / hr and an applied voltage of approximately 13.3 kV. After primary drying and optical microscopy, ImageJ analysis showed a mean particle size of 19.95 μm and a dispersibility index of 0.27 (Figure 11).

[0179] A 3 mL volume of desalted IgG solution was prepared at a concentration of 50 mg / mL by exchanging the PBS buffer solution with deionized water using a concentration column. This solution was electrosprayed at a flow rate of 0.4 mL / hr and an applied voltage of 10.6 kV. After primary drying and SEM imaging, ImageJ showed a mean particle size of 18.06 μm and a dispersibility index of 0.12 (Figure 12). The salt content was found to be less than 1.5 wt% (note that the initial PBS buffer contained approximately 8 g / L NaCl). The particle density and water content after primary drying were 1.324 g / mL and 7-10 wt%, respectively. After performing the secondary drying step, a residual water content of less than 3 wt% was achieved.

[0180] 5 mL of desalted mAb solution was prepared at a concentration of approximately 20 mg / mL by exchanging the buffer solution with deionized water in a concentration column. This solution was electrosprayed at a flow rate of approximately 0.4 mL / hr and an applied voltage of approximately 11.8 kV. After primary drying and optical microscopy, ImageJ analysis showed a mean particle size of 16.02 μm and a dispersity index of 0.09 (Figure 13).

[0181] Aggregate analysis

[0182] A 1 mL solution of mAb1 at a concentration of 20 mg / mL containing an 8 mg / mL additive was prepared at pH 6.5 (feed solution). This solution was electrosprayed at a flow rate of 0.4 mL / hr and an applied voltage of 11.9 kV. Aggregate analysis was performed by HPLC after (i) primary drying, (ii) secondary drying, (iii) and storage under accelerated conditions (7 days at 40°C). The aggregate size distribution was compared to that observed in both the feed solution and the labeled formulation. When stored under similar conditions, after 7 days of accelerated storage, the particle formulation had approximately 0.2% fewer aggregates than the labeled formulation (see Table 2 below).

[0183] [Table 3]

[0184] IgG function by ELISA assay

[0185] A solution of IgG at a concentration of 50 mg / mL was electrosprayed at a flow rate of approximately 0.4 mL / hr and an applied voltage of 10.6 kV. After primary drying, the particles were reconstituted in PBS so that the IgG contained therein could be analyzed by ELISA assay. Figure 15 shows that the signal of the reconstituted IgG was equivalent to that of the control sample in Figure 14, indicating that the binding activity of the Fc (constant) domain of the electrosprayed protein was preserved.

[0186] Cell-binding activity of mAb1 and mAb2 (in vitro indicator of therapeutic activity)

[0187] A solution of mAb1 at a concentration of 20 mg / mL was electrosprayed at a flow rate of approximately 0.4 mL / hr and an applied voltage of 11.8 kV. A second solution of mAb2 at a concentration of 20 mg / mL was electrosprayed at a flow rate of approximately 0.4 mL / hr and an applied voltage of 11.9 kV. The particles were subjected to primary drying. Figures 16 and 17 show the binding activity of the labeled formulations for mAb1 and mAb2, as assessed by flow cytometry. A heat-denatured control and a mAb-negative control, which have no affinity for the target expressed by the cell line, demonstrated that the assay was sensitive to heat denaturation and selectivity. Particles containing mAb1 (Figure 16) and mAb2 (Figure 17) were then reconstituted in PBS and assayed for binding activity. The results indicated that the full binding activity of the antibodies was preserved throughout the electrospray particle formation process.

[0188] In Figure 18, the thermal stability of mAb1 particles was evaluated over 7 days of accelerated storage at 40°C. The particles were split into two samples: one stored as a dry powder and the other stored in benzyl benzoate. After storage, the particles were reconstituted in PBS, at which point flow cytometry assays again showed preservation of cell binding activity compared to untreated material (labeled formulation) without storage.

[0189] Viscosity of IgG particle suspension

[0190] Desalted IgG and mAb1 particles were prepared by electrospray as described above. These particles were individually suspended in benzyl benzoate and analyzed in a rheometer at a shear rate of approximately 1000 s. Both suspensions exhibited similar rheological behavior and performed substantially better than the aqueous formulation of mAb1 (Figure 19). Suspension formulations of either drug above 300 mg / mL were observed to have apparent viscosities of approximately less than 20 cP. Furthermore, IgG particles in a 500 mg / mL suspension were found to have an [η] value of approximately 2.64 dL / g, indicating that the viscosity was primarily dominated by the excluded volume effect. This should be compared with aqueous solutions of various proteins, where intermolecular interactions can contribute significantly to the apparent viscosity of the medium; for example, aqueous IgG is characterized by an [η] value around 6.5 dL / g (E. Longman, SE Harding, N. Mareineke, LCGC North America, 2006, 24, 1, 64-72).

[0191] Comparison of human IgG particles produced with and without an electric field

[0192] Human IgG particles were generated by drying droplets of aqueous IgG aerosol (approximately 16 mg / mL IgG, 4 mg / mL NaCl) generated by an ultrasonic nozzle. Aerosol formation for one group of particles (Group A) was assisted by an electric field applied by biasing 8 kV between the nozzle and an electrode placed several centimeters downstream (Figure 20), whereas it was not assisted for the other group (Group B) (Figure 21).

[0193] The average particle sizes of Groups A and B were 6.5 and 10.1 μm, respectively. This indicates that the electric field acted to reduce the average droplet size. The standard deviations were 1.8 and 3.4 μm, respectively. The particles were then reconstituted in water and analyzed by HPLC. Aggregates were found to be 7.9% (Group A) and 6.9% (Group B) of the protein mass, compared to 14.1% aggregates in the feed solution. The zeta potential of the same reconstituted material was 3.95 mV (Group A) and 6.91 mV (Group B).

[0194] Surface-protruding additives in human IgG particles produced by electrospray

[0195] A solution of approximately 80 mg / mL human IgG and approximately 30 mg / mL salt was electrosprayed at a flow rate of 0.4 mL / hr and an applied voltage of 12.2 kV. After primary drying of the droplets, particles were formed and visualized by SEM (Figures 22 and 23). Large amounts of crystalline or semi-crystalline salt were observed on the surface of the particles.

[0196] A solution of approximately 80 mg / mL human IgG, approximately 20 mg / mL salt, and approximately 10 mg / mL sugar was electrosprayed at a flow rate of 0.4 mL / hr and an applied voltage of 12 kV. After primary drying of the droplets, particles were generated and visualized by SEM (Figures 24 and 25). Salt and / or sugar were observed in large amounts on the surface of the particles.

[0197] A monoclonal antibody solution containing approximately 70 mg / mL mAb3 and approximately 46 mg / mL sugar was electrosprayed at a flow rate of 0.4 mL / hr and an applied voltage of 12.1 kV. After primary drying of the droplets, particles were generated and visualized by SEM (Figures 26 and 27). Sugar was observed in large amounts on the surface of the particles.

[0198] Other embodiments are within the claims.

Claims

1. 1. A method of electrospraying a first liquid containing a first therapeutic or diagnostic agent to form droplets, and evaporating the first liquid to produce particles from the droplets, wherein the therapeutic or diagnostic agent in the particles has an activity of 0.5 to 1.0 activity / unit.

2. 10. The method of claim 1, wherein the concentration of the first therapeutic or diagnostic agent in the first liquid is 0.0001 to 1000 mg / mL.

3. The method of claim 1 or 2, wherein the first liquid is aqueous.

4. 4. The method of claim 3, wherein the aqueous first liquid is selected from the group consisting of water, 0.9% saline, lactated Ringer's solution, dextrose 5%, or a buffer solution.

5. 5. The method of claim 4, wherein the buffer is selected from the group consisting of acetate buffer, histidine buffer, succinate buffer, HEPES buffer, Tris buffer, carbonate buffer, citrate buffer, phosphate buffer, glycine buffer, barbital buffer, and cacodylate buffer.

6. 10. The method of claim 1, wherein the first liquid further comprises carbohydrates, pH adjusters, salts, chelating agents, minerals, polymers, surfactants, protein stabilizers, emulsifiers, preservatives, amino acids, antioxidants, proteins, organic solvents, and nutrient media.

7. 7. The method of claim 6, wherein the carbohydrate is dextran, trehalose, sucrose, agarose, mannitol, lactose, sorbitol, or maltose.

8. 7. The method of claim 6, wherein the pH adjuster is acetate, citrate, glutamate, glycinate, histidine, lactate, maleate, phosphate, succinate, tartrate, bicarbonate, aluminum hydroxide, phosphoric acid, hydrochloric acid, DL-lactic acid / glycolic acid, phosphorylethanolamine, tromethamine, imidazole, glycylglycine, or monosodium glutamate.

9. 7. The method of claim 6, wherein the salt is sodium chloride, calcium chloride, potassium chloride, sodium hydroxide, stannous chloride, magnesium sulfate, sodium glucoheptonate, sodium pertechnetate, or guanidine hydrochloride.

10. 7. The method of claim 6, wherein the chelating agent is edetate disodium.

11. 7. The method of claim 6, wherein the mineral is calcium, zinc, or titanium dioxide.

12. 7. The method of claim 6, wherein the polymer is propylene glycol, glucose star polymer, silicone polymer, polydimethylsiloxane, polyethylene glycol, carboxymethylcellulose, poly(glycolic acid), poly(lactic-co-glycolic acid), or polylactic acid.

13. 7. The method of claim 6, wherein the surfactant is polysorbate, magnesium stearate, sodium dodecyl sulfate, Triton N-101, glycerin, or polyoxyethylated castor oil.

14. 7. The method of claim 6, wherein the protein stabilizer is acetyltryptophan salt, caprylate salt, or N-acetyltryptophan.

15. 7. The method of claim 6, wherein the emulsifier is polysorbate 80, polysorbate 20, sorbitan monooleate, ethanolamine, polyoxyl 35 castor oil, polyoxyl 40 hydrogenated castor oil, carbomer 1342, corn oil-mono-di-triglyceride, polyoxyethylated oleic acid glyceride, or poloxamer.

16. 7. The method of claim 6, wherein the preservative is phenol, m-cresol, benzyl alcohol, 2-phenyloxyethanol, chlorobutanol, neomycin, benzethonium chloride, glutaraldehyde, or beta-propiolactone.

17. 7. The method of claim 6, wherein the amino acid is alanine, aspartic acid, cysteine, isoleucine, glutamic acid, leucine, methionine, phenylalanine, pyrrolidine, serine, selenocysteine, threonine, tryptophan, tyrosine, valine, asparagine, L-arginine, histidine, glycine, or glutamine.

18. 7. The method of claim 6, wherein the antioxidant is glutathione, ascorbic acid, cysteine, or tocopherol.

19. 7. The method of claim 6, wherein the protein is protamine, protamine sulfate, or gelatin.

20. The method of claim 6, wherein the organic solvent is dimethyl sulfoxide or N-methyl-2-pyrrolidone.

21. 7. The method of claim 6, wherein the preservative is methyl hydroxybenzoate, thimerosal, a paraben, formaldehyde, or castor oil.

22. The method of claim 1 , wherein the first liquid further comprises adenine, tri-n-butyl phosphate, octafluoropropane, white petrolatum, or p-aminophenyl-p-anisate.

23. The method of claim 1 or 2, wherein the first liquid is an organic solvent.

24. 24. The method of claim 23, wherein the organic solvent is selected from the group consisting of dichloromethane, dimethyl sulfoxide, urea, sarcosine, methanol, formic acid, acetic acid, ethyl acetate, acetonitrile, acetone, methyl acetate, diethyl ether, hydrazine, ethyl nitrate, butanol, dimethoxyethane, methyl tert-butyl ether, triethylamine, and any combination thereof.

25. 25. The method of any one of claims 1 to 24, wherein the particles have a diameter of 0.1 to 1000 μm.

26. 26. The method of any one of claims 1 to 25, wherein the particles have a polydispersity index of 0.05 to 0.

9.

27. A method according to any one of claims 1 to 26, wherein after electrospraying, the particles are suspended in a non-aqueous or aqueous liquid, thereby forming a suspension.

28. 28. The method of claim 27, wherein the non-aqueous liquid is an organic solvent or an ionic liquid.

29. The organic solvent may be selected from the group consisting of benzyl alcohol, benzyl benzoate, castor oil, coconut oil, corn oil, cottonseed oil, fish oil, grape seed oil, hazelnut oil, hydrogenated palm kernel oil, olive oil, peanut oil, peppermint oil, safflower oil, sesame oil, soybean oil, sunflower oil, vegetable oil, walnut oil, polyethylene glycol, glycofurol, acetone, diglyme, dimethylacetamide, dimethyl isosorbide, dimethyl sulfoxide, ethanol, ethyl acetate, ethyl ether, ethyl lactate, isopropyl acetate, methyl acetate, methyl isobutyl ketone, and methyl tert-butyl ether. ether, N-methylpyrrolidone, perfluorodecalin, 2-pyrrolidone, triglycerides, tetrahydrofurfuryl alcohol, fractionated triglycerides of C8 and C10 vegetable fatty acids, propylene glycol diesters of C8 and C10 saturated vegetable fatty acids, ethyl oleate, ethyl caprate, dibutyl adipate, fatty acid esters, hexanoic acid, octanoic acid, triacetin, diethyl glycol monoether, gamma-butyrolactone, eugenol, clove bud oil, citral, limonene, and any combination thereof; or The ionic liquid may be selected from the group consisting of pyridinium, pyridazinium, pyrimidinium, pyrazinium, imidazolium, pyrazolium, thiazolium, oxazolium, triazolium, ammonium, sulfonium, halide, sulfate, sulfonate, carbonate, phosphate, bicarbonate, nitrate, acetate, PF ６ － , B.F. ４ － , triflate, nonaflate, bis(trifyl)amide, trifluoroacetate, heptafluorobutanoate, haloaluminate, or any combination thereof; 29. The method of claim 28.

30. 28. The method of claim 27, wherein the aqueous liquid is selected from the group consisting of water, 0.9% saline, lactated Ringer's solution, dextrose 5%, or a buffer solution.

31. 31. The method of any one of claims 27 to 30, wherein the suspension further comprises carbohydrates, pH adjusters, salts, chelating agents, minerals, polymers, surfactants, protein stabilizers, emulsifiers, preservatives, amino acids, antioxidants, proteins, organic solvents, and nutrient media.

32. 32. The method of any one of claims 27 to 31, wherein the suspension has a viscosity of 0.27 to 200 cP.

33. 33. The method of any one of claims 27 to 32, wherein the suspension comprises 5 to 90% by volume of particles.

34. 34. The method of any one of claims 27 to 33, wherein the suspension has a concentration of the first therapeutic or diagnostic agent of 0.0001 to 1000 mg / mL.

35. 35. The method of any one of claims 1 to 34, wherein the first therapeutic or diagnostic agent is selected from the group consisting of a nucleic acid, an antibody, a peptide, a protein, a cell, a carbohydrate, a chemical agent, a contrast agent, a magnetic particle, a polymer bead, a metal nanoparticle, a metal microparticle, a quantum dot, an antioxidant, an antibiotic, a hormone, a nucleoprotein, a polysaccharide, a glycoprotein, a lipoprotein, a steroid, an analgesic, a local anesthetic, an anti-inflammatory agent, an antimicrobial agent, a chemotherapeutic agent, an exosome, an outer membrane vesicle, a vaccine, a virus, a bacteriophage, an adjuvant, a vitamin, a mineral, an organelle, and any combination thereof.

36. 36. The method of any one of claims 27 to 35, wherein the non-aqueous or aqueous liquid comprises a second therapeutic or diagnostic agent.

37. 37. The method of claim 36, wherein the first therapeutic or diagnostic agent and the second therapeutic or diagnostic agent are the same.

38. 37. The method of claim 36, wherein the first therapeutic or diagnostic agent and the second therapeutic or diagnostic agent are different.

39. 39. The method of any one of claims 36 to 38, wherein the concentration of the second therapeutic or diagnostic agent in the non-aqueous or aqueous liquid is from 0.0001 to 1000 mg / mL.

40. 40. The method of any one of claims 36 to 39, wherein the second therapeutic or diagnostic agent is selected from the group consisting of a nucleic acid, an antibody, a peptide, a protein, a cell, a carbohydrate, a chemical agent, a contrast agent, a magnetic particle, a polymer bead, a metal nanoparticle, a metal microparticle, a quantum dot, an antioxidant, an antibiotic, a hormone, a nucleoprotein, a polysaccharide, a glycoprotein, a lipoprotein, a steroid, an analgesic, a local anesthetic, an anti-inflammatory agent, an antimicrobial agent, a chemotherapeutic agent, an exosome, an outer membrane vesicle, a vaccine, a virus, a bacteriophage, an adjuvant, a vitamin, a mineral, an organelle, and any combination thereof.

41. 1. A method of administering a first therapeutic or diagnostic agent to a mammal, the method comprising administering to the mammal an effective amount of a suspension or dry formulation comprising particles, the suspension comprising a non-aqueous or aqueous liquid, and the particles comprising the first therapeutic or diagnostic agent, the first therapeutic or diagnostic agent having 0.5 to 1.0 activity / unit.

42. 42. The method of claim 41, wherein the suspension has a viscosity of 0.27 to 200 cP.

43. 43. The method of claim 41 or 42, wherein the suspension comprises 5 to 90% by volume of particles.

44. 44. The method of any one of claims 41 to 43, wherein the suspension has a concentration of the first therapeutic or diagnostic agent of 0.0001 to 1000 mg / mL.

45. 45. The method of any one of claims 41 to 44, wherein the first therapeutic or diagnostic agent is selected from the group consisting of a nucleic acid, an antibody, a peptide, a protein, a cell, a carbohydrate, a chemical agent, a contrast agent, a magnetic particle, a polymer bead, a metal nanoparticle, a metal microparticle, a quantum dot, an antioxidant, an antibiotic, a hormone, a nucleoprotein, a polysaccharide, a glycoprotein, a lipoprotein, a steroid, an analgesic, a local anesthetic, an anti-inflammatory agent, an antimicrobial agent, a chemotherapeutic agent, an exosome, an outer membrane vesicle, a vaccine, a virus, a bacteriophage, an adjuvant, a vitamin, a mineral, an organelle, and any combination thereof.

46. 46. ​​The method of any one of claims 41 to 45, wherein the non-aqueous liquid is an organic solvent or an ionic liquid.

47. The organic solvent may be selected from the group consisting of benzyl alcohol, benzyl benzoate, castor oil, coconut oil, corn oil, cottonseed oil, fish oil, grape seed oil, hazelnut oil, hydrogenated palm kernel oil, olive oil, peanut oil, peppermint oil, safflower oil, sesame oil, soybean oil, sunflower oil, vegetable oil, walnut oil, polyethylene glycol, glycofurol, acetone, diglyme, dimethylacetamide, dimethyl isosorbide, dimethyl sulfoxide, ethanol, ethyl acetate, ethyl ether, ethyl lactate, isopropyl acetate, methyl acetate, methyl isobutyl ketone, and methyl tert-butyl ether. ether, N-methylpyrrolidone, perfluorodecalin, 2-pyrrolidone, triglycerides, tetrahydrofurfuryl alcohol, fractionated triglycerides of C8 and C10 vegetable fatty acids, propylene glycol diesters of C8 and C10 saturated vegetable fatty acids, ethyl oleate, ethyl caprate, dibutyl adipate, fatty acid esters, hexanoic acid, octanoic acid, triacetin, diethyl glycol monoether, gamma-butyrolactone, eugenol, clove bud oil, citral, limonene, and any combination thereof; or The ionic liquid may be selected from the group consisting of pyridinium, pyridazinium, pyrimidinium, pyrazinium, imidazolium, pyrazolium, thiazolium, oxazolium, triazolium, ammonium, sulfonium, halide, sulfate, sulfonate, carbonate, phosphate, bicarbonate, nitrate, acetate, PF ６ － , B.F. ４ － , triflate, nonaflate, bis(trifyl)amide, trifluoroacetate, heptafluorobutanoate, haloaluminate, or any combination thereof; 47. The method of claim 46.

48. 46. ​​The method of any one of claims 41 to 45, wherein the aqueous liquid is selected from the group consisting of water, 0.9% saline, lactated Ringer's solution, dextrose 5%, or a buffer solution.

49. 49. The method of any one of claims 41 to 48, wherein the suspension further comprises carbohydrates, pH adjusters, salts, chelating agents, minerals, polymers, surfactants, protein stabilizers, emulsifiers, preservatives, amino acids, antioxidants, proteins, organic solvents, and nutrient media.

50. 50. The method of any one of claims 41 to 49, wherein the non-aqueous or aqueous liquid comprises a second therapeutic or diagnostic agent.

51. 51. The method of claim 50, wherein the first therapeutic or diagnostic agent and the second therapeutic or diagnostic agent are the same.

52. 51. The method of claim 50, wherein the first therapeutic or diagnostic agent and the second therapeutic or diagnostic agent are different.

53. 53. The method of any one of claims 50 to 52, wherein the concentration of the second therapeutic or diagnostic agent in the non-aqueous or aqueous liquid is from 0.0001 to 1000 mg / mL.

54. 54. The method of any one of claims 50 to 53, wherein the second therapeutic or diagnostic agent is selected from the group consisting of a nucleic acid, an antibody, a peptide, a protein, a cell, a carbohydrate, a chemical agent, a contrast agent, a magnetic particle, a polymer bead, a metal nanoparticle, a metal microparticle, a quantum dot, an antioxidant, an antibiotic, a hormone, a nucleoprotein, a polysaccharide, a glycoprotein, a lipoprotein, a steroid, an analgesic, a local anesthetic, an anti-inflammatory agent, an antimicrobial agent, a chemotherapeutic agent, an exosome, an outer membrane vesicle, a vaccine, a virus, a bacteriophage, an adjuvant, a vitamin, a mineral, an organelle, and any combination thereof.

55. 55. The method of any one of claims 41 to 54, wherein the particles have a diameter of 0.1 to 1000 μm.

56. 56. The method of any one of claims 41 to 55, wherein the particles have a polydispersity index of 0.05 to 0.

9.

57. The suspension or dry formulation may be administered intraauricularly, intrabuccally, conjunctivally, on the skin, in teeth, electroosmotically, intracervically, intrasinusally, intratracheally, enterally, epidurally, extraamniotically, extracorporeally, by infiltration, interstitially, intraperitoneally, intraamniotically, intraarterially, intraarticularly, intrabiliary, intrabronchial, intravesically, intracardially, intracartilaginously, intrasaccularly, intrasaccularly, intracavity, intracavity, intracerebrally, intracisternally, intracornea, intradental crown, intracoronary artery, intracavernosally, intradermally, intradiscally, intraductally, intraduodenum, intradurally, intraepidermally, intraesophageally, intragastrically, intragingivally, intraileum, intralesionally, intraluminally, intralymphatic, intramedullary, intrameningeal, intramuscularly, intraocularly, intraovarianly, intrapericardially, intraperitoneally, intrapleurally, intraprostatically, intrapulmonary, nasal sinus or periorbitally.

57. The method of any one of claims 41-56, wherein the method is administered via intracavernous, intraspinal, intrasynovial, intratendinous, intratesticular, intrathecal, intrathoracic, intratubular, intratumoral, intratympanic, intrauterine, intravascular, intravenous, intravenous bolus, intravenous infusion, intraventricular, intravesical, intravitreal, iontophoretic, lavage, laryngeal, nasal, nasogastric, occlusive dressing, ocular, oral, oropharyngeal, parenteral, transdermal, periarticular, epidural, perineural, periodontal, rectal, inhalation, retrobulbar, soft tissue, subarachnoid, subconjunctival, subcutaneous, sublingual, submucosal, topical, transdermal, transmucosal, transplacental, transtracheal, transtympanic, ureteral, urethral, ​​or vaginal administration.

58. 1. A composition comprising a suspension, said suspension comprising a non-aqueous or aqueous liquid and particles, said particles comprising a first therapeutic or diagnostic agent, said first therapeutic or diagnostic agent having 0.5 to 1.0 activity / unit.

59. 59. The composition of claim 58, wherein the suspension has a viscosity of 0.27 to 200 cP.

60. 60. The composition of claim 58 or 59, wherein the suspension comprises 5 to 90% by volume of particles.

61. 61. The composition of any one of claims 58 to 60, wherein the suspension has a concentration of the first therapeutic or diagnostic agent of 0.0001 to 1000 mg / mL.

62. 62. The composition of any one of claims 58-61, wherein the first therapeutic or diagnostic agent is selected from the group consisting of a nucleic acid, an antibody, a peptide, a protein, a cell, a carbohydrate, a chemical agent, a contrast agent, a magnetic particle, a polymer bead, a metal nanoparticle, a metal microparticle, a quantum dot, an antioxidant, an antibiotic, a hormone, a nucleoprotein, a polysaccharide, a glycoprotein, a lipoprotein, a steroid, an analgesic, a local anesthetic, an anti-inflammatory agent, an antimicrobial agent, a chemotherapeutic agent, an exosome, an outer membrane vesicle, a vaccine, a virus, a bacteriophage, an adjuvant, a vitamin, a mineral, an organelle, and any combination thereof.

63. 63. The composition of any one of claims 58 to 62, wherein the non-aqueous liquid is an organic solvent or an ionic liquid.

64. The organic solvent may be selected from the group consisting of benzyl alcohol, benzyl benzoate, castor oil, coconut oil, corn oil, cottonseed oil, fish oil, grape seed oil, hazelnut oil, hydrogenated palm kernel oil, olive oil, peanut oil, peppermint oil, safflower oil, sesame oil, soybean oil, sunflower oil, vegetable oil, walnut oil, polyethylene glycol, glycofurol, acetone, diglyme, dimethylacetamide, dimethyl isosorbide, dimethyl sulfoxide, ethanol, ethyl acetate, ethyl ether, ethyl lactate, isopropyl acetate, methyl acetate, methyl isobutyl ketone, and methyl tert-butyl ether. ether, N-methylpyrrolidone, perfluorodecalin, 2-pyrrolidone, triglycerides, tetrahydrofurfuryl alcohol, fractionated triglycerides of C8 and C10 vegetable fatty acids, propylene glycol diesters of C8 and C10 saturated vegetable fatty acids, ethyl oleate, ethyl caprate, dibutyl adipate, fatty acid esters, hexanoic acid, octanoic acid, triacetin, diethyl glycol monoether, gamma-butyrolactone, eugenol, clove bud oil, citral, limonene, and any combination thereof; or The ionic liquid may be selected from the group consisting of pyridinium, pyridazinium, pyrimidinium, pyrazinium, imidazolium, pyrazolium, thiazolium, oxazolium, triazolium, ammonium, sulfonium, halide, sulfate, sulfonate, carbonate, phosphate, bicarbonate, nitrate, acetate, PF ６ － , B.F. ４ － , triflate, nonaflate, bis(trifyl)amide, trifluoroacetate, heptafluorobutanoate, haloaluminate, or any combination thereof; 64. The composition of claim 63.

65. 63. The composition of any one of claims 58 to 62, wherein the aqueous liquid is selected from the group consisting of water, 0.9% saline, lactated Ringer's solution, dextrose 5% or a buffer.

66. 66. The composition of any one of claims 58 to 65, wherein the suspension further comprises carbohydrates, pH adjusters, salts, chelating agents, minerals, polymers, surfactants, protein stabilizers, emulsifiers, preservatives, amino acids, antioxidants, proteins, organic solvents, and nutrient media.

67. 67. The composition of any one of claims 58 to 66, wherein the non-aqueous or aqueous liquid comprises a second therapeutic or diagnostic agent.

68. 68. The composition of claim 67, wherein the first therapeutic or diagnostic agent and the second therapeutic or diagnostic agent are the same.

69. 68. The composition of claim 67, wherein the first therapeutic or diagnostic agent and the second therapeutic or diagnostic agent are different.

70. 70. The composition of any one of claims 67 to 69, wherein the concentration of the second therapeutic or diagnostic agent in the non-aqueous or aqueous liquid is from 0.0001 to 1000 mg / mL.

71. 71. The composition of any one of claims 67-70, wherein the second therapeutic or diagnostic agent is selected from the group consisting of a nucleic acid, an antibody, a peptide, a protein, a cell, a carbohydrate, a chemical agent, a contrast agent, a magnetic particle, a polymer bead, a metal nanoparticle, a metal microparticle, a quantum dot, an antioxidant, an antibiotic, a hormone, a nucleoprotein, a polysaccharide, a glycoprotein, a lipoprotein, a steroid, an analgesic, a local anesthetic, an anti-inflammatory agent, an antimicrobial agent, a chemotherapeutic agent, an exosome, an outer membrane vesicle, a vaccine, a virus, a bacteriophage, an adjuvant, a vitamin, a mineral, an organelle, and any combination thereof.

72. 72. The composition of any one of claims 58 to 71, wherein the particles have a diameter of 0.1 to 1000 μm.

73. 73. The composition of any one of claims 58 to 72, wherein the particles have a polydispersity index of 0.05 to 0.

9.

74. A composition comprising particles made by the method of any one of claims 1 to 40.

75. 75. The composition of claim 74, wherein the particles are in suspension in a non-aqueous or aqueous liquid.

76. 76. The composition of claim 75, wherein the suspension has a viscosity of 0.27 to 200 cP.

77. 77. The composition of claim 75 or 76, wherein the suspension comprises 5 to 90% by volume of particles.

78. 78. The composition of any one of claims 74 to 77, wherein the particles comprise the first therapeutic or diagnostic agent at a concentration of 0.0001 to 1000 mg / mL.

79. 79. The composition of any one of claims 74-78, wherein the first therapeutic or diagnostic agent is selected from the group consisting of a nucleic acid, an antibody, a peptide, a protein, a cell, a carbohydrate, a chemical agent, a contrast agent, a magnetic particle, a polymer bead, a metal nanoparticle, a metal microparticle, a quantum dot, an antioxidant, an antibiotic, a hormone, a nucleoprotein, a polysaccharide, a glycoprotein, a lipoprotein, a steroid, an analgesic, a local anesthetic, an anti-inflammatory agent, an antimicrobial agent, a chemotherapeutic agent, an exosome, an outer membrane vesicle, a vaccine, a virus, a bacteriophage, an adjuvant, a vitamin, a mineral, an organelle, and any combination thereof.

80. 80. The composition of any one of claims 75 to 79, wherein the non-aqueous liquid is an organic solvent or an ionic liquid.

81. The organic solvent may be selected from the group consisting of benzyl alcohol, benzyl benzoate, castor oil, coconut oil, corn oil, cottonseed oil, fish oil, grape seed oil, hazelnut oil, hydrogenated palm kernel oil, olive oil, peanut oil, peppermint oil, safflower oil, sesame oil, soybean oil, sunflower oil, vegetable oil, walnut oil, polyethylene glycol, glycofurol, acetone, diglyme, dimethylacetamide, dimethyl isosorbide, dimethyl sulfoxide, ethanol, ethyl acetate, ethyl ether, ethyl lactate, isopropyl acetate, methyl acetate, methyl isobutyl ketone, and methyl tert-butyl ether. ether, N-methylpyrrolidone, perfluorodecalin, 2-pyrrolidone, triglycerides, tetrahydrofurfuryl alcohol, fractionated triglycerides of C8 and C10 vegetable fatty acids, propylene glycol diesters of C8 and C10 saturated vegetable fatty acids, ethyl oleate, ethyl caprate, dibutyl adipate, fatty acid esters, hexanoic acid, octanoic acid, triacetin, diethyl glycol monoether, gamma-butyrolactone, eugenol, clove bud oil, citral, limonene, and any combination thereof; or The ionic liquid may be selected from the group consisting of pyridinium, pyridazinium, pyrimidinium, pyrazinium, imidazolium, pyrazolium, thiazolium, oxazolium, triazolium, ammonium, sulfonium, halide, sulfate, sulfonate, carbonate, phosphate, bicarbonate, nitrate, acetate, PF ６ － , B.F. ４ － , triflate, nonaflate, bis(trifyl)amide, trifluoroacetate, heptafluorobutanoate, haloaluminate, or any combination thereof; 81. The composition of claim 80.

82. 80. The composition of any one of claims 75 to 79, wherein the aqueous liquid is selected from the group consisting of water, 0.9% saline, lactated Ringer's solution, dextrose 5%, or a buffer solution.

83. 83. The composition of any one of claims 75 to 82, wherein the suspension further comprises carbohydrates, pH adjusters, salts, chelating agents, minerals, polymers, surfactants, protein stabilizers, emulsifiers, preservatives, amino acids, antioxidants, proteins, organic solvents, and nutrient media.

84. 84. The composition of any one of claims 75 to 83, wherein the non-aqueous or aqueous liquid comprises a second therapeutic or diagnostic agent.

85. 85. The composition of claim 84, wherein the first therapeutic or diagnostic agent and the second therapeutic or diagnostic agent are the same.

86. 85. The composition of claim 84, wherein the first therapeutic or diagnostic agent and the second therapeutic or diagnostic agent are different.

87. 87. The composition of any one of claims 84 to 86, wherein the concentration of the second therapeutic or diagnostic agent in the non-aqueous or aqueous liquid is 0.0001 to 1000 mg / mL.

88. 88. The composition of any one of claims 84-87, wherein the second therapeutic or diagnostic agent is selected from the group consisting of a nucleic acid, an antibody, a peptide, a protein, a cell, a carbohydrate, a chemical agent, a contrast agent, a magnetic particle, a polymer bead, a metal nanoparticle, a metal microparticle, a quantum dot, an antioxidant, an antibiotic, a hormone, a nucleoprotein, a polysaccharide, a glycoprotein, a lipoprotein, a steroid, an analgesic, a local anesthetic, an anti-inflammatory agent, an antimicrobial agent, a chemotherapeutic agent, an exosome, an outer membrane vesicle, a vaccine, a virus, a bacteriophage, an adjuvant, a vitamin, a mineral, an organelle, and any combination thereof.

89. 89. The composition of any one of claims 74 to 88, wherein the particles have a diameter of 0.1 to 1000 μm.

90. 90. The composition of any one of claims 74 to 89, wherein the particles have a polydispersity index of 0.05 to 0.

9.

91. 91. The composition of any one of claims 74 to 90, wherein the first liquid used to create the particles is aqueous.

92. 92. The composition of claim 91, wherein the aqueous first liquid is selected from the group consisting of water, 0.9% saline, lactated Ringer's solution, dextrose 5%, or a buffer solution.

93. 93. The composition of claim 92, wherein the buffer is selected from the group consisting of acetate buffer, histidine buffer, succinate buffer, HEPES buffer, Tris buffer, carbonate buffer, citrate buffer, phosphate buffer, glycine buffer, barbital buffer, and cacodylate buffer.

94. 75. The composition of claim 74, wherein the first liquid further comprises carbohydrates, pH adjusters, salts, chelating agents, minerals, polymers, surfactants, protein stabilizers, emulsifiers, preservatives, amino acids, antioxidants, proteins, organic solvents, and nutrient media.

95. 95. The composition of claim 94, wherein the carbohydrate is dextran, trehalose, sucrose, agarose, mannitol, lactose, sorbitol, or maltose.

96. 95. The composition of claim 94, wherein the pH adjuster is acetate, citrate, glutamate, glycinate, histidine, lactate, maleate, phosphate, succinate, tartrate, bicarbonate, aluminum hydroxide, phosphoric acid, hydrochloric acid, DL-lactic acid / glycolic acid, phosphorylethanolamine, tromethamine, imidazole, glycylglycine, or monosodium glutamate.

97. 95. The composition of claim 94, wherein the salt is sodium chloride, calcium chloride, potassium chloride, sodium hydroxide, stannous chloride, magnesium sulfate, sodium glucoheptonate, sodium pertechnetate, or guanidine hydrochloride.

98. 95. The composition of claim 94, wherein the chelating agent is edetate disodium.

99. 95. The composition of claim 94, wherein the mineral is calcium, zinc, or titanium dioxide.

100. 95. The composition of claim 94, wherein the polymer is propylene glycol, a glucose star polymer, a silicone polymer, polydimethylsiloxane, polyethylene glycol, carboxymethylcellulose, poly(glycolic acid), poly(lactic-co-glycolic acid), or polylactic acid.

101. 95. The composition of claim 94, wherein the surfactant is polysorbate, magnesium stearate, sodium dodecyl sulfate, Triton N-101, glycerin, or polyoxyethylated castor oil.

102. 95. The composition of claim 94, wherein the protein stabilizer is an acetyltryptophan salt, caprylate salt, or N-acetyltryptophan.

103. 95. The composition of claim 94, wherein the emulsifier is polysorbate 80, polysorbate 20, sorbitan monooleate, ethanolamine, polyoxyl 35 castor oil, polyoxyl 40 hydrogenated castor oil, carbomer 1342, corn oil-mono-di-triglyceride, polyoxyethylated oleic acid glyceride, or poloxamer.

104. 95. The composition of claim 94, wherein the preservative is phenol, m-cresol, benzyl alcohol, 2-phenyloxyethanol, chlorobutanol, neomycin, benzethonium chloride, glutaraldehyde, or beta-propiolactone.

105. 95. The composition of claim 94, wherein the amino acid is alanine, aspartic acid, cysteine, isoleucine, glutamic acid, leucine, methionine, phenylalanine, pyrrolidine, serine, selenocysteine, threonine, tryptophan, tyrosine, valine, asparagine, L-arginine, histidine, glycine, or glutamine.

106. 95. The composition of claim 94, wherein the antioxidant is glutathione, ascorbic acid, cysteine, or tocopherol.

107. 95. The composition of claim 94, wherein the protein is protamine, protamine sulfate, or gelatin.

108. 95. The composition of claim 94, wherein the organic solvent is dimethyl sulfoxide or N-methyl-2-pyrrolidone.

109. 95. The composition of claim 94, wherein the preservative is methyl hydroxybenzoate, thimerosal, a paraben, formaldehyde, or castor oil.

110. 75. The composition of claim 74, wherein the first liquid further comprises adenine, tri-n-butyl phosphate, octafluoropropane, white petrolatum, or p-aminophenyl-p-anisate.

111. 75. The composition of claim 74, wherein the first liquid used to create the particles is an organic solvent.

112. 112. The composition of claim 111, wherein the organic solvent is selected from the group consisting of dichloromethane, dimethyl sulfoxide, urea, sarcosine, methanol, formic acid, acetic acid, ethyl acetate, acetonitrile, acetone, methyl acetate, diethyl ether, hydrazine, ethyl nitrate, butanol, dimethoxyethane, methyl tert-butyl ether, triethylamine, and any combination thereof.

113. 41. A method of administering a first therapeutic or diagnostic agent by administering a composition comprising particles made by the method of any one of claims 1 to 40.

114. 114. The method of claim 113, wherein the composition is a suspension of the particles in a non-aqueous or aqueous liquid.

115. 115. The method of claim 114, wherein the suspension has a viscosity of 0.27 to 200 cP.

116. 116. The method of claim 114 or 115, wherein the suspension comprises 5 to 90% by volume of particles.

117. 117. The method of any one of claims 113 to 116, wherein the concentration of the first therapeutic or diagnostic agent is 0.0001 to 1000 mg / mL.

118. 118. The method of any one of claims 113 to 117, wherein the first therapeutic or diagnostic agent is selected from the group consisting of a nucleic acid, an antibody, a peptide, a protein, a cell, a carbohydrate, a chemical agent, a contrast agent, a magnetic particle, a polymer bead, a metal nanoparticle, a metal microparticle, a quantum dot, an antioxidant, an antibiotic, a hormone, a nucleoprotein, a polysaccharide, a glycoprotein, a lipoprotein, a steroid, an analgesic, a local anesthetic, an anti-inflammatory agent, an antimicrobial agent, a chemotherapeutic agent, an exosome, an outer membrane vesicle, a vaccine, a virus, a bacteriophage, an adjuvant, a vitamin, a mineral, an organelle, and any combination thereof.

119. 119. The method of any one of claims 114 to 118, wherein the non-aqueous liquid is an organic solvent or an ionic liquid.

120. The organic solvent may be selected from the group consisting of benzyl alcohol, benzyl benzoate, castor oil, coconut oil, corn oil, cottonseed oil, fish oil, grape seed oil, hazelnut oil, hydrogenated palm kernel oil, olive oil, peanut oil, peppermint oil, safflower oil, sesame oil, soybean oil, sunflower oil, vegetable oil, walnut oil, polyethylene glycol, glycofurol, acetone, diglyme, dimethylacetamide, dimethyl isosorbide, dimethyl sulfoxide, ethanol, ethyl acetate, ethyl ether, ethyl lactate, isopropyl acetate, methyl acetate, methyl isobutyl ketone, and methyl tert-butyl ether. ether, N-methylpyrrolidone, perfluorodecalin, 2-pyrrolidone, triglycerides, tetrahydrofurfuryl alcohol, fractionated triglycerides of C8 and C10 vegetable fatty acids, propylene glycol diesters of C8 and C10 saturated vegetable fatty acids, ethyl oleate, ethyl caprate, dibutyl adipate, fatty acid esters, hexanoic acid, octanoic acid, triacetin, diethyl glycol monoether, gamma-butyrolactone, eugenol, clove bud oil, citral, limonene, and any combination thereof; or The ionic liquid may be selected from the group consisting of pyridinium, pyridazinium, pyrimidinium, pyrazinium, imidazolium, pyrazolium, thiazolium, oxazolium, triazolium, ammonium, sulfonium, halide, sulfate, sulfonate, carbonate, phosphate, bicarbonate, nitrate, acetate, PF ６ － , B.F. ４ － , triflate, nonaflate, bis(trifyl)amide, trifluoroacetate, heptafluorobutanoate, haloaluminate, or any combination thereof; 120. The method of claim 119.

121. 119. The method of any one of claims 114 to 118, wherein the aqueous liquid is selected from the group consisting of water, 0.9% saline, lactated Ringer's solution, dextrose 5%, or a buffer solution.

122. 122. The method of any one of claims 114 to 121, wherein the suspension further comprises carbohydrates, pH adjusters, salts, chelating agents, minerals, polymers, surfactants, protein stabilizers, emulsifiers, preservatives, amino acids, antioxidants, proteins, organic solvents, and nutrient media.

123. 123. The method of any one of claims 114 to 122, wherein the non-aqueous or aqueous liquid comprises a second therapeutic or diagnostic agent.

124. 124. The method of claim 123, wherein the first therapeutic or diagnostic agent and the second therapeutic or diagnostic agent are the same.

125. 124. The method of claim 123, wherein the first therapeutic or diagnostic agent and the second therapeutic or diagnostic agent are different.

126. 126. The method of any one of claims 123 to 125, wherein the concentration of the second therapeutic or diagnostic agent in the non-aqueous or aqueous liquid is from 0.0001 to 1000 mg / mL.

127. 127. The method of any one of claims 123 to 126, wherein the second therapeutic or diagnostic agent is selected from the group consisting of a nucleic acid, an antibody, a peptide, a protein, a cell, a carbohydrate, a chemical agent, a contrast agent, a magnetic particle, a polymer bead, a metal nanoparticle, a metal microparticle, a quantum dot, an antioxidant, an antibiotic, a hormone, a nucleoprotein, a polysaccharide, a glycoprotein, a lipoprotein, a steroid, an analgesic, a local anesthetic, an anti-inflammatory agent, an antimicrobial agent, a chemotherapeutic agent, an exosome, an outer membrane vesicle, a vaccine, a virus, a bacteriophage, an adjuvant, a vitamin, a mineral, an organelle, and any combination thereof.

128. 128. The method of any one of claims 113 to 127, wherein the particles have a diameter of 0.1 to 1000 μm.

129. 129. The method of any one of claims 113 to 128, wherein the particles have a polydispersity index of 0.05 to 0.

9.

130. 130. The method of any one of claims 113 to 129, wherein the first liquid used to create the particles is aqueous.

131. 131. The method of claim 130, wherein the aqueous first liquid is selected from the group consisting of water, 0.9% saline, lactated Ringer's solution, dextrose 5%, or a buffer solution.

132. 132. The method of claim 131, wherein the buffer is selected from the group consisting of acetate buffer, histidine buffer, succinate buffer, HEPES buffer, Tris buffer, carbonate buffer, citrate buffer, phosphate buffer, glycine buffer, barbital buffer, and cacodylate buffer.

133. 114. The method of claim 113, wherein the first liquid further comprises carbohydrates, pH adjusters, salts, chelating agents, minerals, polymers, surfactants, protein stabilizers, emulsifiers, preservatives, amino acids, antioxidants, proteins, organic solvents, and nutrient media.

134. 134. The method of claim 133, wherein the carbohydrate is dextran, trehalose, sucrose, agarose, mannitol, lactose, sorbitol, or maltose.

135. 134. The method of claim 133, wherein the pH adjuster is acetate, citrate, glutamate, glycinate, histidine, lactate, maleate, phosphate, succinate, tartrate, bicarbonate, aluminum hydroxide, phosphoric acid, hydrochloric acid, DL-lactic acid / glycolic acid, phosphorylethanolamine, tromethamine, imidazole, glycylglycine, or monosodium glutamate.

136. 134. The method of claim 133, wherein the salt is sodium chloride, calcium chloride, potassium chloride, sodium hydroxide, stannous chloride, magnesium sulfate, sodium glucoheptonate, sodium pertechnetate, or guanidine hydrochloride.

137. 134. The method of claim 133, wherein the chelating agent is edetate disodium.

138. 134. The method of claim 133, wherein the mineral is calcium, zinc, or titanium dioxide.

139. 134. The method of claim 133, wherein the polymer is propylene glycol, a glucose star polymer, a silicone polymer, polydimethylsiloxane, polyethylene glycol, carboxymethylcellulose, poly(glycolic acid), poly(lactic-co-glycolic acid), or polylactic acid.

140. 134. The method of claim 133, wherein the surfactant is polysorbate, magnesium stearate, sodium dodecyl sulfate, Triton N-101, glycerin, or polyoxyethylated castor oil.

141. 134. The method of claim 133, wherein the protein stabilizing agent is an acetyltryptophan salt, caprylate salt, or N-acetyltryptophan.

142. 134. The method of claim 133, wherein the emulsifier is polysorbate 80, polysorbate 20, sorbitan monooleate, ethanolamine, polyoxyl 35 castor oil, polyoxyl 40 hydrogenated castor oil, carbomer 1342, corn oil-mono-di-triglyceride, polyoxyethylated oleic acid glyceride, or poloxamer.

143. 134. The method of claim 133, wherein the preservative is phenol, m-cresol, benzyl alcohol, 2-phenyloxyethanol, chlorobutanol, neomycin, benzethonium chloride, glutaraldehyde, or beta-propiolactone.

144. 134. The method of claim 133, wherein the amino acid is alanine, aspartic acid, cysteine, isoleucine, glutamic acid, leucine, methionine, phenylalanine, pyrrolysine, serine, selenocysteine, threonine, tryptophan, tyrosine, valine, asparagine, L-arginine, histidine, glycine, or glutamine.

145. 134. The method of claim 133, wherein the antioxidant is glutathione, ascorbic acid, cysteine, or tocopherol.

146. 134. The method of claim 133, wherein the protein is protamine, protamine sulfate, or gelatin.

147. 134. The method of claim 133, wherein the organic solvent is dimethyl sulfoxide or N-methyl-2-pyrrolidone.

148. 134. The method of claim 133, wherein the preservative is methyl hydroxybenzoate, thimerosal, a paraben, formaldehyde, or castor oil.

149. 114. The method of claim 113, wherein the first liquid further comprises adenine, tri-n-butyl phosphate, octafluoropropane, white petrolatum, or p-aminophenyl-p-anisate.

150. 150. The method of any one of claims 113 to 149, wherein the first liquid used to create the particles is an organic solvent.

151. 151. The method of claim 150, wherein the organic solvent is selected from the group consisting of dichloromethane, dimethyl sulfoxide, urea, sarcosine, methanol, formic acid, acetic acid, ethyl acetate, acetonitrile, acetone, methyl acetate, diethyl ether, hydrazine, ethyl nitrate, butanol, dimethoxyethane, methyl tert-butyl ether, triethylamine, and any combination thereof.

152. the composition is delivered to the auricle, buccal, conjunctival, skin, teeth, electroosmotic, intracervical, intrasinus, intratracheal, enteral, epidural, extra-amniotic, extracorporeal, infiltrative, interstitial, intraperitoneal, intra-amniotic, intra-arterial, intra-articular, intra-biliary, intra-bronchial, intra-vesical, intracardiac, intra-cartilage, intra-sacral, intra-cavity, intra-cavity, intra-cerebral, intra-cisternal, intra-corneal, intra-dental, intra-coronary, intra-cavernous, intra-dermal, intra-discal, intra-ductal, intra-duodenal, intra-dural, intra-epidermal, intra-esophageal, intra-gastric, intra-gingival, intra-ileal, intra-lesional, intra-luminal, intra-lymphatic, intra-bone marrow, intra-meningeal, intra-muscular, intra-ocular, intra-ovarian, intra-pericardium, intra-peritoneum, intra-pleural, intra-prostate, intra-pulmonary, nasal or peri-orbital sinus, 152. The method of any one of claims 113-151, wherein the method is administered by intraspinal, intrasynovial, intratendinous, intratesticular, intrathecal, intrathoracic, intratubular, intratumoral, intratympanic, intrauterine, intravascular, intravenous, intravenous bolus, intravenous infusion, intraventricular, intravesical, intravitreal, iontophoretic, lavage, laryngeal, nasal, nasogastric, occlusive dressing, ocular, oral, oropharyngeal, parenteral, transdermal, periarticular, epidural, perineural, periodontal, rectal, inhalation, retrobulbar, soft tissue, subarachnoid, subconjunctival, subcutaneous, sublingual, submucosal, topical, transdermal, transmucosal, transplacental, transtracheal, transtympanic, ureteral, urethral, ​​or vaginal administration.

153. 1. A method of forming particles by electrospraying an annular flow of an encapsulating material toward a collector, and electrospraying a flow of a liquid toward the collector centrally to the annular flow of the encapsulating material, the flow comprising a first therapeutic or diagnostic agent, wherein the particles are collected at the collector, and the concentration of the first therapeutic or diagnostic agent in the liquid ranges from 1 to 1000 mg / mL.

154. 154. The method of claim 153, wherein the particles are formulated into a pharmaceutical composition in dry form or suspended in a pharmaceutically acceptable vehicle.

155. 155. The method of claim 154, wherein the encapsulating material is selected from the group consisting of poly(vinyl alcohol), poly(acrylic acid), poly(acrylamide), poly(ethylene oxide), poly(lactic acid), poly(glycolic acid), polycaprolactone, poly(lactic-co-glycolic acid), chitosan, cellulose, and any combination thereof.

156. 156. The method of claim 154 or 155, wherein the liquid is an organic solvent.

157. The organic solvent may be selected from the group consisting of benzyl alcohol, benzyl benzoate, castor oil, coconut oil, corn oil, cottonseed oil, fish oil, grape seed oil, hazelnut oil, hydrogenated palm kernel oil, olive oil, peanut oil, peppermint oil, safflower oil, sesame oil, soybean oil, sunflower oil, vegetable oil, walnut oil, polyethylene glycol, glycofurol, acetone, diglyme, dimethylacetamide, dimethyl isosorbide, dimethyl sulfoxide, ethanol, ethyl acetate, ethyl ether, ethyl lactate, isopropyl acetate, methyl acetate, methyl isobutyl ketone, methyl tert-butyl ether, N- 157. The method of claim 156, wherein the surfactant is selected from the group consisting of methylpyrrolidone, perfluorodecalin, 2-pyrrolidone, triglycerides, tetrahydrofurfuryl alcohol, fractionated triglycerides of C8 and C10 vegetable fatty acids, propylene glycol diesters of C8 and C10 saturated vegetable fatty acids, ethyl oleate, ethyl caprate, dibutyl adipate, fatty acid esters, hexanoic acid, octanoic acid, triacetin, diethyl glycol monoether, gamma-butyrolactone, eugenol, clove bud oil, citral, limonene, and any combination thereof.

158. 156. The method of any one of claims 154 or 155, wherein the liquid is aqueous.

159. 159. The method of claim 158, wherein the aqueous liquid is selected from the group consisting of water, 0.9% saline, lactated Ringer's solution, or a buffer solution.

160. 160. The method of claim 159, wherein the buffer is selected from the group consisting of acetate buffer, histidine buffer, succinate buffer, HEPES buffer, Tris buffer, carbonate buffer, citrate buffer, phosphate buffer, glycine buffer, barbital buffer, and cacodylate buffer.

161. 154. The method of claim 153, wherein the liquid further comprises a carbohydrate, a pH adjuster, a salt, a chelating agent, a mineral, a polymer, a surfactant, a protein stabilizer, an emulsifier, a preservative, an amino acid, an antioxidant, a protein, an organic solvent, or a nutrient medium.

162. 162. The method of claim 161, wherein the carbohydrate is dextran, trehalose, sucrose, agarose, mannitol, lactose, sorbitol, or maltose.

163. 162. The method of claim 161, wherein the pH adjuster is acetate, citrate, glutamate, glycinate, histidine, lactate, maleate, phosphate, succinate, tartrate, bicarbonate, aluminum hydroxide, phosphoric acid, hydrochloric acid, DL-lactic acid / glycolic acid, phosphorylethanolamine, tromethamine, imidazole, glycylglycine, or monosodium glutamate.

164. 162. The method of claim 161, wherein the salt is sodium chloride, calcium chloride, potassium chloride, sodium hydroxide, stannous chloride, magnesium sulfate, sodium glucoheptonate, sodium pertechnetate, or guanidine hydrochloride.

165. 162. The method of claim 161, wherein the chelating agent is edetate disodium.

166. 162. The method of claim 161, wherein the mineral is calcium, zinc, or titanium dioxide.

167. 162. The method of claim 161, wherein the polymer is propylene glycol, glucose star polymer, silicone polymer, polydimethylsiloxane, polyethylene glycol, carboxymethylcellulose, poly(glycolic acid), poly(lactic-co-glycolic acid), or polylactic acid.

168. 162. The method of claim 161, wherein the surfactant is polysorbate, magnesium stearate, sodium dodecyl sulfate, polyethylene glycol nonylphenyl ether, glycerin, or polyoxyethylated castor oil.

169. 162. The method of claim 161, wherein the protein stabilizing agent is acetyltryptophan salt, caprylate salt, or N-acetyltryptophan.

170. 162. The method of claim 161, wherein the emulsifier is polysorbate 80, polysorbate 20, sorbitan monooleate, ethanolamine, polyoxyl 35 castor oil, polyoxyl 40 hydrogenated castor oil, carbomer 1342, corn oil-mono-di-triglyceride, polyoxyethylated oleic acid glyceride, or poloxamer.

171. 162. The method of claim 161, wherein the preservative is phenol, m-cresol, benzyl alcohol, 2-phenyloxyethanol, chlorobutanol, neomycin, benzethonium chloride, glutaraldehyde, or beta-propiolactone.

172. 162. The method of claim 161, wherein the amino acid is alanine, aspartic acid, cysteine, isoleucine, glutamic acid, leucine, methionine, phenylalanine, pyrrolysine, serine, selenocysteine, threonine, tryptophan, tyrosine, valine, asparagine, L-arginine, histidine, glycine, or glutamine.

173. 162. The method of claim 161, wherein the antioxidant is glutathione, ascorbic acid, cysteine, or tocopherol.

174. 162. The method of claim 161, wherein the protein is protamine, protamine sulfate, or gelatin.

175. 162. The method of claim 161, wherein the organic solvent is dimethyl sulfoxide or N-methyl-2-pyrrolidone.

176. 162. The method of claim 161, wherein the preservative is methyl hydroxybenzoate, thimerosal, a paraben, formaldehyde, or castor oil.

177. 155. The method of claim 154, wherein the liquid further comprises adenine, tri-n-butyl phosphate, octafluoropropane, white petrolatum, or p-aminophenyl-p-anisate.

178. 156. The method of any one of claims 154 or 155, wherein the liquid is an ionic liquid.

179. The ionic liquid may be selected from the group consisting of pyridinium, pyridazinium, pyrimidinium, pyrazinium, imidazolium, pyrazolium, thiazolium, oxazolium, triazolium, ammonium, sulfonium, halide, sulfate, sulfonate, carbonate, phosphate, bicarbonate, nitrate, acetate, PF ６ － , B.F. ４ － , triflate, nonaflate, bis(trifyl)amide, trifluoroacetate, heptafluorobutanoate, haloaluminate, or any combination thereof.

180. 156. The method of any one of claims 154 or 155, wherein the liquid is a hydrogel or an ionogel.

181. 181. The method of claim 180, wherein the hydrogel or ionogel is selected from the group consisting of collagen hydrogels, chitosan hydrogels, methylcellulose hydrogels, dextran hydrogels, alginate hydrogels, agarose hydrogels, poly(methyl methacrylate) hydrogels, poly(amidoamine) hydrogels, poly(ethyleneimine) hydrogels, polyethylene oxide hydrogels, gelatin hydrogels, hyaluronic acid hydrogels, and any combination thereof.

182. 182. The method of any one of claims 154 to 181, wherein the pharmaceutical composition has a concentration of the first therapeutic or diagnostic agent of 0.0001 to 1000 mg / mL.

183. 183. The method of any one of claims 153-182, wherein the first therapeutic or diagnostic agent is selected from the group consisting of a nucleic acid, an antibody, a peptide, a protein, a cell, a carbohydrate, a chemical agent, a contrast agent, a magnetic particle, a polymer bead, a metal nanoparticle, a metal microparticle, a quantum dot, an antioxidant, an antibiotic, a hormone, a nucleoprotein, a polysaccharide, a glycoprotein, a lipoprotein, a steroid, an analgesic, a local anesthetic, an anti-inflammatory agent, an antimicrobial agent, a chemotherapeutic agent, an exosome, an outer membrane vesicle, a vaccine, a virus, a bacteriophage, an adjuvant, a vitamin, a mineral, an organelle, and any combination thereof.

184. 184. The method of any one of claims 154 to 183, wherein the pharmaceutical composition comprises a second therapeutic or diagnostic agent.

185. 185. The method of claim 184, wherein the first therapeutic or diagnostic agent and the second therapeutic or diagnostic agent are the same.

186. 185. The method of claim 184, wherein the first therapeutic or diagnostic agent and the second therapeutic or diagnostic agent are different.

187. 187. The method of any one of claims 184 to 186, wherein the concentration of the second therapeutic or diagnostic agent in the pharmaceutical composition is 0.0001 to 1000 mg / mL.

188. 188. The method of any one of claims 184-187, wherein the second therapeutic or diagnostic agent is selected from the group consisting of a nucleic acid, an antibody, a peptide, a protein, a cell, a carbohydrate, a chemical agent, a contrast agent, a magnetic particle, a polymer bead, a metal nanoparticle, a metal microparticle, a quantum dot, an antioxidant, an antibiotic, a hormone, a nucleoprotein, a polysaccharide, a glycoprotein, a lipoprotein, a steroid, an analgesic, a local anesthetic, an anti-inflammatory agent, an antimicrobial agent, a chemotherapeutic agent, an exosome, an outer membrane vesicle, a vaccine, a virus, a bacteriophage, an adjuvant, a vitamin, a mineral, an organelle, and any combination thereof.

189. 189. The method of any one of claims 154 to 188, wherein the medium is an organic solvent.

190. The organic solvent may be selected from the group consisting of benzyl alcohol, benzyl benzoate, castor oil, coconut oil, corn oil, cottonseed oil, fish oil, grape seed oil, hazelnut oil, hydrogenated palm kernel oil, olive oil, peanut oil, peppermint oil, safflower oil, sesame oil, soybean oil, sunflower oil, vegetable oil, walnut oil, polyethylene glycol, glycofurol, acetone, diglyme, dimethylacetamide, dimethyl isosorbide, dimethyl sulfoxide, ethanol, ethyl acetate, ethyl ether, ethyl lactate, isopropyl acetate, methyl acetate, methyl isobutyl ketone, methyl tert-butyl ether, N- 190. The method of claim 189, wherein the surfactant is selected from the group consisting of methylpyrrolidone, perfluorodecalin, 2-pyrrolidone, triglycerides, tetrahydrofurfuryl alcohol, fractionated C8 and C10 triglycerides of vegetable fatty acids, propylene glycol diesters of saturated C8 and C10 vegetable fatty acids, ethyl oleate, ethyl caprate, dibutyl adipate, fatty acid esters, hexanoic acid, octanoic acid, triacetin, diethyl glycol monoether, gamma-butyrolactone, eugenol, clove bud oil, citral, limonene, and any combination thereof.

191. 189. The method of any one of claims 154 to 188, wherein the medium is aqueous.

192. 192. The method of claim 191, wherein the aqueous medium is selected from the group consisting of water, 0.9% saline, lactated Ringer's solution, or a buffer solution.

193. 193. The method of claim 192, wherein the buffer is selected from the group consisting of acetate buffer, histidine buffer, succinate buffer, HEPES buffer, Tris buffer, carbonate buffer, citrate buffer, phosphate buffer, glycine buffer, barbital buffer, and cacodylate buffer.

194. 155. The method of claim 154, wherein the medium further comprises a carbohydrate, a pH adjuster, a salt, a chelating agent, a mineral, a polymer, a surfactant, a protein stabilizer, an emulsifier, a preservative, an amino acid, an antioxidant, a protein, an organic solvent, or a nutrient medium.

195. 195. The method of claim 194, wherein the carbohydrate is dextran, trehalose, sucrose, agarose, mannitol, lactose, sorbitol, or maltose.

196. 195. The method of claim 194, wherein the pH adjuster is acetate, citrate, glutamate, glycinate, histidine, lactate, maleate, phosphate, succinate, tartrate, bicarbonate, aluminum hydroxide, phosphoric acid, hydrochloric acid, DL-lactic acid / glycolic acid, phosphorylethanolamine, tromethamine, imidazole, glycylglycine, or monosodium glutamate.

197. 195. The method of claim 194, wherein the salt is sodium chloride, calcium chloride, potassium chloride, sodium hydroxide, stannous chloride, magnesium sulfate, sodium glucoheptonate, sodium pertechnetate, or guanidine hydrochloride.

198. 195. The method of claim 194, wherein the chelating agent is edetate disodium.

199. 195. The method of claim 194, wherein the mineral is calcium, zinc, or titanium dioxide.

200. 200. The method of claim 194, wherein the polymer is propylene glycol, glucose star polymer, silicone polymer, polydimethylsiloxane, polyethylene glycol, carboxymethylcellulose, poly(glycolic acid), poly(lactic-co-glycolic acid), or polylactic acid.

201. 195. The method of claim 194, wherein the surfactant is polysorbate, magnesium stearate, sodium dodecyl sulfate, polyethylene glycol nonylphenyl ether, glycerin, or polyoxyethylated castor oil.

202. 195. The method of claim 194, wherein the protein stabilizing agent is an acetyltryptophan salt, caprylate salt, or N-acetyltryptophan.

203. 195. The method of claim 194, wherein the emulsifier is polysorbate 80, polysorbate 20, sorbitan monooleate, ethanolamine, polyoxyl 35 castor oil, polyoxyl 40 hydrogenated castor oil, carbomer 1342, corn oil-mono-di-triglyceride, polyoxyethylated oleic acid glyceride, or poloxamer.

204. 195. The method of claim 194, wherein the preservative is phenol, m-cresol, benzyl alcohol, 2-phenyloxyethanol, chlorobutanol, neomycin, benzethonium chloride, glutaraldehyde, or beta-propiolactone.

205. 195. The method of claim 194, wherein the amino acid is alanine, aspartic acid, cysteine, isoleucine, glutamic acid, leucine, methionine, phenylalanine, pyrrolidine, serine, selenocysteine, threonine, tryptophan, tyrosine, valine, asparagine, L-arginine, histidine, glycine, or glutamine.

206. 195. The method of claim 194, wherein the antioxidant is glutathione, ascorbic acid, cysteine, or tocopherol.

207. 195. The method of claim 194, wherein the protein is protamine, protamine sulfate, or gelatin.

208. 195. The method of claim 194, wherein the organic solvent is dimethyl sulfoxide or N-methyl-2-pyrrolidone.

209. 195. The method of claim 194, wherein the preservative is methyl hydroxybenzoate, thimerosal, a paraben, formaldehyde, or castor oil.

210. 193. The method of claim 192, wherein the aqueous medium further comprises adenine, tri-n-butyl phosphate, octafluoropropane, white petrolatum, or p-aminophenyl-p-anisate.

211. 189. The method of any one of claims 154 to 188, wherein the medium is an ionic liquid.

212. The ionic liquid may be selected from the group consisting of pyridinium, pyridazinium, pyrimidinium, pyrazinium, imidazolium, pyrazolium, thiazolium, oxazolium, triazolium, ammonium, sulfonium, halide, sulfate, sulfonate, carbonate, phosphate, bicarbonate, nitrate, acetate, PF ６ － , B.F. ４ － , triflate, nonaflate, bis(trifyl)amide, trifluoroacetate, heptafluorobutanoate, haloaluminate, or any combination thereof.

213. 189. The method of any one of claims 154 to 188, wherein the medium is a hydrogel or an ionogel.

214. 214. The method of claim 213, wherein the hydrogel or ionogel is selected from the group consisting of collagen hydrogels, chitosan hydrogels, methylcellulose hydrogels, dextran hydrogels, alginate hydrogels, agarose hydrogels, poly(methyl methacrylate) hydrogels, poly(amidoamine) hydrogels, poly(ethyleneimine) hydrogels, polyethylene oxide hydrogels, gelatin hydrogels, hyaluronic acid hydrogels, and any combination thereof.

215. 215. The method of any one of claims 154 to 214, wherein the particles have a diameter of 0.1 to 1000 μm.

216. 216. The method of any one of claims 154 to 215, wherein the particles have a polydispersity index of 0.05 to 0.

9.

217. 217. The method of any one of claims 154 to 216, wherein the liquid has a viscosity of 1 to 5000 cP.

218. 218. The method of any one of claims 154 to 217, wherein the pharmaceutical composition has a viscosity of 0.27 to 200 cP.

219. 219. The method of any one of claims 154 to 218, wherein the pharmaceutical composition comprises 5 to 90% by volume of particles.

220. 1. A method of administering a first therapeutic or diagnostic agent to a mammal, said method comprising administering to said mammal an effective amount of a pharmaceutical composition, said pharmaceutical composition comprising a pharmaceutically acceptable vehicle and particles, said particles comprising said therapeutic or diagnostic agent, said pharmaceutical composition having a viscosity of 0.27 to 200 cP and a concentration of said first therapeutic or diagnostic agent of 5 to 1000 mg / mL, or said pharmaceutical composition comprising particles in dry form comprising said therapeutic or diagnostic agent.

221. 221. The method of claim 220, wherein the pharmaceutical composition comprises 5 to 90% by volume of particles.

222. 222. The method of claim 220 or 221, wherein the pharmaceutical composition has a concentration of the first therapeutic or diagnostic agent of 100 to 1000 mg / mL.

223. 223. The method of any one of claims 220-222, wherein the first therapeutic or diagnostic agent is selected from the group consisting of a nucleic acid, an antibody, a peptide, a protein, a cell, a carbohydrate, a chemical agent, a contrast agent, a magnetic particle, a polymer bead, a metal nanoparticle, a metal microparticle, a quantum dot, an antioxidant, an antibiotic, a hormone, a nucleoprotein, a polysaccharide, a glycoprotein, a lipoprotein, a steroid, an analgesic, a local anesthetic, an anti-inflammatory agent, an antimicrobial agent, a chemotherapeutic agent, an exosome, an outer membrane vesicle, a vaccine, a virus, a bacteriophage, an adjuvant, a vitamin, a mineral, an organelle, and any combination thereof.

224. 224. The method of any one of claims 220 to 223, wherein the pharmaceutical composition comprises a second therapeutic or diagnostic agent.

225. 225. The method of claim 224, wherein the first therapeutic or diagnostic agent and the second therapeutic or diagnostic agent are the same.

226. 225. The method of claim 224, wherein the first therapeutic or diagnostic agent and the second therapeutic or diagnostic agent are different.

227. 227. The method of any one of claims 224 to 226, wherein the concentration of the second therapeutic or diagnostic agent in the pharmaceutical composition is 0.0001 to 1000 mg / mL.

228. 228. The method of any one of claims 224-227, wherein the second therapeutic or diagnostic agent is selected from the group consisting of a nucleic acid, an antibody, a peptide, a protein, a cell, a carbohydrate, a chemical agent, a contrast agent, a magnetic particle, a polymer bead, a metal nanoparticle, a metal microparticle, a quantum dot, an antioxidant, an antibiotic, a hormone, a nucleoprotein, a polysaccharide, a glycoprotein, a lipoprotein, a steroid, an analgesic, a local anesthetic, an anti-inflammatory agent, an antimicrobial agent, a chemotherapeutic agent, an exosome, an outer membrane vesicle, a vaccine, a virus, a bacteriophage, an adjuvant, a vitamin, a mineral, an organelle, and any combination thereof.

229. 229. The method of any one of claims 220 to 228, wherein the medium is an organic solvent.

230. The organic solvent may be selected from the group consisting of benzyl alcohol, benzyl benzoate, castor oil, coconut oil, corn oil, cottonseed oil, fish oil, grape seed oil, hazelnut oil, hydrogenated palm kernel oil, olive oil, peanut oil, peppermint oil, safflower oil, sesame oil, soybean oil, sunflower oil, vegetable oil, walnut oil, polyethylene glycol, glycofurol, acetone, diglyme, dimethylacetamide, dimethyl isosorbide, dimethyl sulfoxide, ethanol, ethyl acetate, ethyl ether, ethyl lactate, isopropyl acetate, methyl acetate, methyl isobutyl ketone, methyl tert-butyl ether, N- 230. The method of claim 229, wherein the surfactant is selected from the group consisting of methylpyrrolidone, perfluorodecalin, 2-pyrrolidone, triglycerides, tetrahydrofurfuryl alcohol, fractionated triglycerides of C8 and C10 vegetable fatty acids, propylene glycol diesters of C8 and C10 saturated vegetable fatty acids, ethyl oleate, ethyl caprate, dibutyl adipate, fatty acid esters, hexanoic acid, octanoic acid, triacetin, diethyl glycol monoether, gamma-butyrolactone, eugenol, clove bud oil, citral, limonene, and any combination thereof.

231. 229. The method of any one of claims 220 to 228, wherein the medium is aqueous.

232. 232. The method of claim 231, wherein the aqueous medium is selected from the group consisting of water, 0.9% saline, lactated Ringer's solution, or a buffer solution.

233. 233. The method of claim 232, wherein the buffer is selected from the group consisting of acetate buffer, histidine buffer, succinate buffer, HEPES buffer, Tris buffer, carbonate buffer, citrate buffer, phosphate buffer, glycine buffer, barbital buffer, and cacodylate buffer.

234. 221. The method of claim 220, wherein the medium further comprises a carbohydrate, a pH adjuster, a salt, a chelating agent, a mineral, a polymer, a surfactant, a protein stabilizer, an emulsifier, a preservative, an amino acid, an antioxidant, a protein, an organic solvent, or a nutrient medium.

235. 235. The method of claim 234, wherein the carbohydrate is dextran, trehalose, sucrose, agarose, mannitol, lactose, sorbitol, or maltose.

236. 235. The method of claim 234, wherein the pH adjuster is acetate, citrate, glutamate, glycinate, histidine, lactate, maleate, phosphate, succinate, tartrate, bicarbonate, aluminum hydroxide, phosphoric acid, hydrochloric acid, DL-lactic acid / glycolic acid, phosphorylethanolamine, tromethamine, imidazole, glycylglycine, or monosodium glutamate.

237. 235. The method of claim 234, wherein the salt is sodium chloride, calcium chloride, potassium chloride, sodium hydroxide, stannous chloride, magnesium sulfate, sodium glucoheptonate, sodium pertechnetate, or guanidine hydrochloride.

238. 235. The method of claim 234, wherein the chelating agent is edetate disodium.

239. 235. The method of claim 234, wherein the mineral is calcium, zinc, or titanium dioxide.

240. 235. The method of claim 234, wherein the polymer is propylene glycol, glucose star polymer, silicone polymer, polydimethylsiloxane, polyethylene glycol, carboxymethylcellulose, poly(glycolic acid), poly(lactic-co-glycolic acid), or polylactic acid.

241. 235. The method of claim 234, wherein the surfactant is polysorbate, magnesium stearate, sodium dodecyl sulfate, polyethylene glycol nonylphenyl ether, glycerin, or polyoxyethylated castor oil.

242. 235. The method of claim 234, wherein the protein stabilizer is an acetyltryptophan salt, caprylate salt, or N-acetyltryptophan.

243. 235. The method of claim 234, wherein the emulsifier is polysorbate 80, polysorbate 20, sorbitan monooleate, ethanolamine, polyoxyl 35 castor oil, polyoxyl 40 hydrogenated castor oil, carbomer 1342, corn oil-mono-di-triglyceride, polyoxyethylated oleic acid glyceride, or poloxamer.

244. 235. The method of claim 234, wherein the preservative is phenol, m-cresol, benzyl alcohol, 2-phenyloxyethanol, chlorobutanol, neomycin, benzethonium chloride, glutaraldehyde, or beta-propiolactone.

245. 235. The method of claim 234, wherein the amino acid is alanine, aspartic acid, cysteine, isoleucine, glutamic acid, leucine, methionine, phenylalanine, pyrrolysine, serine, selenocysteine, threonine, tryptophan, tyrosine, valine, asparagine, L-arginine, histidine, glycine, or glutamine.

246. 235. The method of claim 234, wherein the antioxidant is glutathione, ascorbic acid, cysteine, or tocopherol.

247. 235. The method of claim 234, wherein the protein is protamine, protamine sulfate, or gelatin.

248. 235. The method of claim 234, wherein the organic solvent is dimethyl sulfoxide or N-methyl-2-pyrrolidone.

249. 235. The method of claim 234, wherein the preservative is methyl hydroxybenzoate, thimerosal, a paraben, formaldehyde, or castor oil.

250. 221. The method of claim 220, wherein the medium further comprises adenine, tri-n-butyl phosphate, octafluoropropane, white petrolatum, or p-aminophenyl-p-anisate.

251. 221. The method of claim 220, wherein the medium is an ionic liquid.

252. The ionic liquid may be selected from the group consisting of pyridinium, pyridazinium, pyrimidinium, pyrazinium, imidazolium, pyrazolium, thiazolium, oxazolium, triazolium, ammonium, sulfonium, halide, sulfate, sulfonate, carbonate, phosphate, bicarbonate, nitrate, acetate, PF ６ － , B.F. ４ － , triflate, nonaflate, bis(trifyl)amide, trifluoroacetate, heptafluorobutanoate, haloaluminate, or any combination thereof.

253. 221. The method of claim 220, wherein the medium is a hydrogel or an ionogel.

254. 254. The method of claim 253, wherein the hydrogel or ionogel is selected from the group consisting of collagen hydrogels, chitosan hydrogels, methylcellulose hydrogels, dextran hydrogels, alginate hydrogels, agarose hydrogels, poly(methyl methacrylate) hydrogels, poly(amidoamine) hydrogels, poly(ethyleneimine) hydrogels, polyethylene oxide hydrogels, gelatin hydrogels, hyaluronic acid hydrogels, and any combination thereof.

255. 255. The method of any one of claims 220 to 254, wherein the particles have a diameter of 0.1 to 1000 μm.

256. 256. The method of any one of claims 220 to 255, wherein the particles have a polydispersity index of 0.05 to 0.

9.

257. the pharmaceutical composition is administered intravenously to a patient in the following locations: auricle, buccal, conjunctival, skin, teeth, electroosmotic, intracervical, intrasinus, intratracheal, enteral, epidural, extraamniotic, extracorporeal, infiltrative, interstitial, intraperitoneal, intraamniotic, intra-arterial, intra-articular, intra-biliary, intra-bronchial, intra-vesical, intracardiac, intracartilage, intrasacral, intra-cavity, intra-cavity, intracerebral, intra-cisternal, intra-corneal, intra-coronary, intra-coronary, intra-cavernosal, intradermal, intra-discal, intra-ductal, intra-duodenal, intra-dural, intra-epidermal, intra-esophageal, intra-gastric, intra-gingival, intra-ileum, intra-lesion, intra-luminal, intra-lymphatic, intra-bone marrow, intra-meningeal, intra-muscular, intra-ocular, intra-ovarian, intra-pericardium, intra-peritoneum, intra-pleural, intra-prostate, intra-pulmonary, nasal or peri-orbital sinus , intraspinal, intrasynovial, intratendinous, intratesticular, intrathecal, intrathoracic, intratubular, intratumoral, intratympanic, intrauterine, intravascular, intravenous, intravenous bolus, intravenous infusion, intraventricular, intravesical, intravitreal, iontophoretic, lavage, laryngeal, nasal, nasogastric, occlusive dressing, ocular, oral, oropharyngeal, parenteral, transdermal, periarticular, epidural, perineural, periodontal, rectal, inhalation, retrobulbar, soft tissue, subarachnoid, subconjunctival, subcutaneous, sublingual, submucosal, topical, transdermal, transmucosal, transplacental, transtracheal, transtympanic, ureteral, urethral, ​​and vaginal administration.

258. 1. A composition comprising a vehicle and particles, wherein the particles comprise a first therapeutic or diagnostic agent, and wherein the pharmaceutical composition has a viscosity of 0.27-200 cP and a concentration of the first therapeutic or diagnostic agent of 0.5-1000 mg / mL, or wherein the pharmaceutical composition comprises particles in dry form comprising the therapeutic or diagnostic agent.

259. The composition of claim 258, wherein the composition comprises 5 to 90% by volume of particles.

260. 260. The composition of claim 258 or 259, wherein the composition has a concentration of the first therapeutic or diagnostic agent of 100 to 1000 mg / mL.

261. 261. The composition of any one of claims 258-260, wherein the first therapeutic or diagnostic agent is selected from the group consisting of a nucleic acid, an antibody, a peptide, a protein, a cell, a carbohydrate, a chemical agent, a contrast agent, a magnetic particle, a polymer bead, a metal nanoparticle, a metal microparticle, a quantum dot, an antioxidant, an antibiotic, a hormone, a nucleoprotein, a polysaccharide, a glycoprotein, a lipoprotein, a steroid, an analgesic, a local anesthetic, an anti-inflammatory agent, an antimicrobial agent, a chemotherapeutic agent, an exosome, an outer membrane vesicle, a vaccine, a virus, a bacteriophage, an adjuvant, a vitamin, a mineral, an organelle, and any combination thereof.

262. 262. The composition of any one of claims 258-261, wherein the pharmaceutical composition comprises a second therapeutic or diagnostic agent.

263. 263. The composition of claim 262, wherein the first therapeutic or diagnostic agent and the second therapeutic or diagnostic agent are the same.

264. 263. The composition of claim 262, wherein the first therapeutic or diagnostic agent and the second therapeutic or diagnostic agent are different.

265. 265. The composition of any one of claims 262-264, wherein the concentration of the second therapeutic or diagnostic agent in the pharmaceutical composition is 0.0001 to 1000 mg / mL.

266. 266. The composition of any one of claims 262-265, wherein the second therapeutic or diagnostic agent is selected from the group consisting of a nucleic acid, an antibody, a peptide, a protein, a cell, a carbohydrate, a chemical agent, a contrast agent, a magnetic particle, a polymer bead, a metal nanoparticle, a metal microparticle, a quantum dot, an antioxidant, an antibiotic, a hormone, a nucleoprotein, a polysaccharide, a glycoprotein, a lipoprotein, a steroid, an analgesic, a local anesthetic, an anti-inflammatory agent, an antimicrobial agent, a chemotherapeutic agent, an exosome, an outer membrane vesicle, a vaccine, a virus, a bacteriophage, an adjuvant, a vitamin, a mineral, an organelle, and any combination thereof.

267. The composition of any one of claims 258 to 266, wherein the medium is an organic solvent.

268. The organic solvent may be selected from the group consisting of benzyl alcohol, benzyl benzoate, castor oil, coconut oil, corn oil, cottonseed oil, fish oil, grape seed oil, hazelnut oil, hydrogenated palm kernel oil, olive oil, peanut oil, peppermint oil, safflower oil, sesame oil, soybean oil, sunflower oil, vegetable oil, walnut oil, polyethylene glycol, glycofurol, acetone, diglyme, dimethylacetamide, dimethyl isosorbide, dimethyl sulfoxide, ethanol, ethyl acetate, ethyl ether, ethyl lactate, isopropyl acetate, methyl acetate, methyl isobutyl ketone, methyl tert-butyl ether, N-methyl 268. The composition of claim 267, wherein the fatty acid esters are selected from the group consisting of ethyl pyrrolidone, perfluorodecalin, 2-pyrrolidone, triglycerides, tetrahydrofurfuryl alcohol, fractionated triglycerides of C8 and C10 vegetable fatty acids, propylene glycol diesters of C8 and C10 saturated vegetable fatty acids, ethyl oleate, ethyl caprate, dibutyl adipate, fatty acid esters, hexanoic acid, octanoic acid, triacetin, diethyl glycol monoether, gamma-butyrolactone, eugenol, clove bud oil, citral, limonene, and any combination thereof.

269. The composition of any one of claims 258 to 266, wherein the medium is aqueous.

270. 270. The composition of claim 269, wherein the aqueous vehicle is selected from the group consisting of water, 0.9% saline, lactated Ringer's solution, or a buffer solution.

271. The composition of claim 270, wherein the buffer is selected from the group consisting of acetate buffer, histidine buffer, succinate buffer, HEPES buffer, Tris buffer, carbonate buffer, citrate buffer, phosphate buffer, glycine buffer, barbital buffer, and cacodylate buffer.

272. The composition of claim 258, wherein the vehicle further comprises a carbohydrate, a pH adjuster, a salt, a chelating agent, a mineral, a polymer, a surfactant, a protein stabilizer, an emulsifier, a preservative, an amino acid, an antioxidant, a protein, an organic solvent, or a nutrient medium.

273. 273. The composition of claim 272, wherein the carbohydrate is dextran, trehalose, sucrose, agarose, mannitol, lactose, sorbitol, or maltose.

274. The composition of claim 272, wherein the pH adjuster is acetate, citrate, glutamate, glycinate, histidine, lactate, maleate, phosphate, succinate, tartrate, bicarbonate, aluminum hydroxide, phosphoric acid, hydrochloric acid, DL-lactic acid / glycolic acid, phosphorylethanolamine, tromethamine, imidazole, glycylglycine, or monosodium glutamate.

275. The composition of claim 272, wherein the salt is sodium chloride, calcium chloride, potassium chloride, sodium hydroxide, stannous chloride, magnesium sulfate, sodium glucoheptonate, sodium pertechnetate, or guanidine hydrochloride.

276. The composition of claim 272, wherein the chelating agent is edetate disodium.

277. 273. The composition of claim 272, wherein the mineral is calcium, zinc, or titanium dioxide.

278. 273. The composition of claim 272, wherein the polymer is propylene glycol, glucose star polymer, silicone polymer, polydimethylsiloxane, polyethylene glycol, carboxymethylcellulose, poly(glycolic acid), poly(lactic-co-glycolic acid), or polylactic acid.

279. The composition of claim 272, wherein the surfactant is polysorbate, magnesium stearate, sodium dodecyl sulfate, polyethylene glycol nonylphenyl ether, glycerin, or polyoxyethylated castor oil.

280. 273. The composition of claim 272, wherein the protein stabilizer is an acetyltryptophan salt, caprylate salt, or N-acetyltryptophan.

281. 273. The composition of claim 272, wherein the emulsifier is polysorbate 80, polysorbate 20, sorbitan monooleate, ethanolamine, polyoxyl 35 castor oil, polyoxyl 40 hydrogenated castor oil, carbomer 1342, corn oil-mono-di-triglyceride, polyoxyethylated oleic acid glyceride, or poloxamer.

282. The composition of claim 272, wherein the preservative is phenol, m-cresol, benzyl alcohol, 2-phenyloxyethanol, chlorobutanol, neomycin, benzethonium chloride, glutaraldehyde, or beta-propiolactone.

283. 273. The composition of claim 272, wherein the amino acid is alanine, aspartic acid, cysteine, isoleucine, glutamic acid, leucine, methionine, phenylalanine, pyrrolidine, serine, selenocysteine, threonine, tryptophan, tyrosine, valine, asparagine, L-arginine, histidine, glycine, or glutamine.

284. 273. The composition of claim 272, wherein the antioxidant is glutathione, ascorbic acid, cysteine, or tocopherol.

285. 273. The composition of claim 272, wherein the protein is protamine, protamine sulfate, or gelatin.

286. 273. The composition of claim 272, wherein the organic solvent is dimethyl sulfoxide or N-methyl-2-pyrrolidone.

287. The composition of claim 272, wherein the preservative is methyl hydroxybenzoate, thimerosal, a paraben, formaldehyde, or castor oil.

288. The composition of claim 258, wherein the vehicle further comprises adenine, tri-n-butyl phosphate, octafluoropropane, white petrolatum, or p-aminophenyl-p-anisate.

289. The composition of claim 258, wherein the medium is an ionic liquid.

290. The ionic liquid may be selected from the group consisting of pyridinium, pyridazinium, pyrimidinium, pyrazinium, imidazolium, pyrazolium, thiazolium, oxazolium, triazolium, ammonium, sulfonium, halide, sulfate, sulfonate, carbonate, phosphate, bicarbonate, nitrate, acetate, PF ６ － , B.F. ４ － , triflate, nonaflate, bis(trifyl)amide, trifluoroacetate, heptafluorobutanoate, haloaluminate, or any combination thereof.

291. The composition of claim 258, wherein the medium is a hydrogel or an ionogel.

292. 292. The composition of claim 291, wherein the hydrogel or ionogel is selected from the group consisting of collagen hydrogels, chitosan hydrogels, methylcellulose hydrogels, dextran hydrogels, alginate hydrogels, agarose hydrogels, poly(methyl methacrylate) hydrogels, poly(amidoamine) hydrogels, poly(ethyleneimine) hydrogels, polyethylene oxide hydrogels, gelatin hydrogels, hyaluronic acid hydrogels, and any combination thereof.

293. 293. The composition of any one of claims 258 to 292, wherein the particles have a diameter of 0.1 to 1000 μm.

294. 294. The composition of any one of claims 258 to 293, wherein the particles have a polydispersity index of 0.05 to 0.

9.

295. 220. A composition comprising particles made by the method of any one of claims 153 to 219.

296. The composition of claim 295, wherein the particles are in a pharmaceutically acceptable medium, thereby forming a pharmaceutical composition.

297. 220. A method of administering a first therapeutic or diagnostic agent by administering a pharmaceutical composition comprising particles made by the method of any one of claims 154 to 219.

298. The composition may be administered intravenously through an auricle, buccal, conjunctival, skin, tooth, electroosmotic, intracervical, intrasinus, intratracheal, enteral, epidural, extraamniotic, extracorporeal, infiltrative, interstitial, intraperitoneal, intraamniotic, intra-arterial, intra-articular, intra-biliary, intra-bronchial, intra-vesical, intracardiac, intracartilaginous, intrasacral, intra-cavity, intra-cavity, intracerebral, intra-cisternal, intra-corneal, intra-coronary, intra-coronary, intra-cavernosal, intradermal, intra-discal, intra-ductal, intra-duodenal, intra-dural, intra-epidermal, intra-esophageal, intra-gastric, intra-gingival, intra-ileal, intra-lesional, intra-luminal, intra-lymphatic, intra-medullary, intra-meningeal, intra-muscular, intra-ocular, intra-ovarian, intra-pericardial, intra-peritoneal, intra-pleural, intra-prostatic, intra-pulmonary, nasal sinus, or ocular 298. The method of claim 297, wherein the administration is via perifobic sinus, intraspinal, intrasynovial, intratendinous, intratesticular, intrathecal, intrathoracic, intrarenal tubule, intratumor, intratympanic, intrauterine, intravascular, intravenous, intravenous bolus, intravenous infusion, intraventricular, intravesical, intravitreal, iontophoretic, lavage, laryngeal, nasal, nasogastric, occlusive dressing, ocular, oral, oropharyngeal, parenteral, transdermal, periarticular, epidural, perineural, periodontal, rectal, inhalation, retrobulbar, soft tissue, subarachnoid, subconjunctival, subcutaneous, sublingual, submucosal, topical, transdermal, transmucosal, transplacental, transtracheal, transtympanic, ureteral, urethral, ​​and vaginal administration.

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