Methods and systems for forming particles comprising therapeutics

Abstract

The present disclosure relates to methods of forming particles comprising a therapeutic agent, the methods comprising: a) providing a first liquid stream comprising an aqueous feed solution, the feed solution comprising the therapeutic agent; b) providing a second liquid stream comprising an organic liquid; c) dispersing the first liquid stream into the second liquid stream at a junction; d) applying turbulent flow conditions to a fluid channel connected to the junction, the fluid channel containing the first liquid stream and the second liquid stream, to form aqueous droplets comprising the therapeutic agent; e) dehydrating an aqueous fraction of the aqueous droplets, the dehydrating comprising contacting the droplets with a dehydration liquid to form particles comprising the therapeutic agent; and f) collecting the particles aseptically.

Description

5726.1016001METHODS AND SYSTEMS FOR FORMING PARTICLES COMPRISING THERAPEUTICS RELATED APPLICATION(S)

[0001] This application claims the benefit of U.S. Provisional Application No.63 / 721,342, filed on November 15, 2024. The entire teachings of the above application(s) are incorporated herein by reference.BACKGROUND

[0002] Efficient, reproducible, and scalable methods are needed to produce particles that may be incorporated into drug products for research or commercial use. The present disclosure relates to processes and equipment that can be used to manufacture large quantities of drug products comprising particles having certain desirable quality attributes.SUMMARY

[0003] Provided herein are methods allowing aqueous liquid droplet formation, dehydration of the aqueous droplets, and removal of an aqueous fraction of the aqueous droplets to produce particles comprising a therapeutic agent.

[0004] In some embodiments, the present disclosure provides a method of making a suspension of particles comprising a therapeutic agent, the method comprising:a) providing a first liquid stream comprising an aqueous feed solution, the feed solution comprising the therapeutic agent;b) providing a second liquid stream comprising an organic liquid;c) dispersing the first liquid stream into the second liquid stream at a junction; d) applying turbulent flow conditions to a fluid channel connected to the junction, the fluid channel containing the first liquid stream and the second liquid stream, to form aqueous droplets comprising the therapeutic agent; e) dehydrating an aqueous fraction of the aqueous droplets, the dehydrating comprising contacting the aqueous droplets with a dehydration liquid to form particles comprising the therapeutic agent;f) removing the dehydration liquid; andg) suspending the particles in a pharmaceutically acceptable carrier liquid to form the suspension.14248642. vl5726.1016001

[0005] In certain embodiments, the organic liquid and the dehydration liquid are the same liquid. In some embodiments, the organic liquid and the dehydration liquid are different liquids. In some embodiments, the dehydration liquid is an organic solvent.

[0006] In some embodiments, the aqueous feed solution further comprises an excipient and the particles further comprise the excipient.

[0007] In other embodiments, the present disclosure provides a method of making a suspension of particles comprising a therapeutic agent, the method comprising:a) providing a first liquid stream comprising an aqueous feed solution, the feed solution comprising the therapeutic agent;b) providing a second liquid stream comprising an organic liquid;c) dispersing the first liquid stream into the second liquid stream at a junction; d) applying turbulent flow conditions to a fluid channel connected to the junction, the fluid channel containing the first liquid stream and the second liquid stream, to form aqueous droplets comprising the therapeutic agent; e) dehydrating an aqueous fraction of the aqueous droplets, the dehydrating comprising contacting the droplets with a dehydration liquid to form particles comprising the therapeutic agent; andf) suspending the particles in a carrier liquid to form the suspension of particles aseptically.

[0008] In some embodiments, a method of adjusting the water content of a plurality of particles in a suspension, the method comprising performing sparging on the suspension, wherein the suspension comprises the particles suspended in a carrier liquid, and wherein the plurality of the particles comprise a therapeutic agent.

[0009] The present disclosure also provides a method of forming particles comprising a therapeutic agent, the method comprising:a) providing a first liquid stream comprising an aqueous feed solution, the feed solution comprising the therapeutic agent;b) providing a second liquid stream comprising an organic liquid;c) dispersing the first liquid stream into the second liquid stream at a first junction;d) applying turbulent flow conditions to a fluid channel connected to the first junction and a second junction, the fluid channel containing the first liquid stream and the second liquid stream, to form aqueous droplets comprising the therapeutic agent;24248642. vl5726.1016001e) dehydrating an aqueous fraction of the aqueous droplets, the dehydrating comprising contacting the droplets with a dehydration liquid to form particles comprising the therapeutic agent; andf) collecting the particles aseptically.

[0010] In some embodiments, a method of forming particles comprising a therapeutic agent is provided herein, wherein the method comprises:a) providing a first liquid stream comprising an aqueous feed solution comprising the therapeutic agent;b) providing a second liquid stream comprising an organic liquid;c) dispersing the first liquid stream into the second liquid stream at or after a first junction;d) applying turbulent flow conditions to a fluid channel connected to the junction, the fluid channel containing the first liquid stream and the second liquid stream, to form aqueous droplets comprising the therapeutic agent; e) dehydrating an aqueous fraction of the aqueous droplets, the dehydrating comprising contacting the droplets with a dehydration liquid at a second junction to form particles comprising the therapeutic agent, wherein the Peclet number of the combination of the first liquid stream, the second liquid stream and the dehydration liquid determines the morphology of the particles; and f) collecting the particles, thereby forming particles comprising the therapeutic agent, wherein the particles have a circularity of from about 0.80 to about 1.00.

[0011] The present disclosure further provides a method of adjusting the volume-to-weight percent water content of particles in a suspension comprising particles in a carrier liquid, the method comprising performing sparging on the suspension, wherein a plurality of the particles comprise at least one therapeutic agent.

[0012] Also disclosed herein is a method of making a suspension of particles comprising a therapeutic agent, the method comprising:a) providing a first liquid stream comprising an aqueous feed solution, the feed solution comprising the therapeutic agent;b) providing a second liquid stream comprising an organic liquid;c) dispersing the first liquid stream into the second liquid stream at a first junction;34248642. vl5726.1016001d) applying turbulent flow conditions to a fluid channel connected to the first junction and a second junction, the fluid channel containing the first liquid stream and the second liquid stream, to form aqueous droplets comprising the therapeutic agent;e) dehydrating an aqueous fraction of the aqueous droplets, the dehydrating comprising contacting the droplets with a dehydration liquid to form particles comprising the therapeutic agent;f) exchanging the dehydration liquid with a carrier liquid aseptically to form the suspension of particles; andg) sparging the suspension of particles to reach a predetermined volume-to- weight percent water content in the particles.BRIEF DESCRIPTION OF THE DRAWINGS

[0013] The foregoing will be apparent from the following more particular description of example embodiments, as illustrated in the accompanying drawings in which like reference characters, refer to the same parts throughout the different views. The drawings are not necessarily to scale, emphasis instead being placed upon illustrating embodiments.

[0014] FIG. 1 illustrates a block diagram of an example of a method for generating particles according to the present disclosure.

[0015] FIG. 2A shows an image of morphology of microparticles generated using a particle-in-tube (PIT) method.

[0016] FIG. 2B shows a size distribution of microparticles generated using a particle-in-tube (PIT) method. The x-axis of the bar chart of FIG. 2B has units of pm.

[0017] FIG. 3 A shows a concentration calibration curve of two suspensions.

[0018] FIG. 3B shows NMR relaxation plot of a suspension and paste with respect to time.

[0019] FIG. 3C shows an NMR plot of a fresh suspension and a resuspension with respect to shaking time.

[0020] FIG. 3D shows an NMR plot of a fresh suspension and three resuspensions (users 1-3) with respect to shaking time.DETAILED DESCRIPTION

[0021] The present disclosure generally relates to methods of making a suspension of particles comprising a therapeutic agent, the methods comprising a) providing a first liquid44248642. vl5726.1016001stream comprising an aqueous feed solution, the feed solution comprising the therapeutic agent; b) providing a second liquid stream comprising an organic liquid; c) dispersing the first liquid stream into the second liquid stream at a junction; d) applying turbulent flow conditions to a fluid channel coupled (e.g., fluidly connected to) to the junction, the fluid channel containing the first liquid stream and the second liquid stream, to form aqueous droplets comprising the therapeutic agent; e) dehydrating an aqueous fraction of the aqueous droplets, the dehydrating comprising contacting the aqueous droplets with a dehydration liquid to form particles comprising the therapeutic agent; f) removing the dehydration liquid; and g) suspending the particles in a pharmaceutically acceptable carrier liquid to form the suspension.

[0022] In some embodiments, the suspension has a volume-to-weight percent water content of less than about 20% (i.e., less than 20 % w / v).

[0023] In some embodiments, step f) comprises exchanging the dehydration liquid for a pharmaceutically acceptable carrier liquid without forming a powder intermediate. In other embodiments, step f) comprises forming a powder intermediate and suspending the powder intermediate in a pharmaceutically acceptable carrier liquid.

[0024] In some embodiments, the first liquid stream further comprises one or more excipients (e.g., an excipient) and the particles further comprise the one or more excipients.

[0025] In certain embodiments, the organic liquid and the dehydration liquid are the same liquid. In some embodiments, the organic liquid and the dehydration liquid are different liquids.

[0026] In certain aspects, the present disclosure generally relates to methods of making a suspension of particles comprising a therapeutic agent, the methods comprising: a) providing a first liquid stream comprising an aqueous feed solution, the feed solution comprising the therapeutic agent; b) providing a second liquid stream comprising an organic liquid; c) dispersing the first liquid stream into the second liquid stream at a junction; d) applying turbulent flow conditions to a fluid channel connected to the junction, the fluid channel containing the first liquid stream and the second liquid stream, to form aqueous droplets comprising the therapeutic agent; e) dehydrating an aqueous fraction of the aqueous droplets, the dehydrating comprising contacting the droplets with a dehydration liquid to form particles comprising the therapeutic agent; and f) suspending the particles in a carrier liquid to form the suspension of particles aseptically.

[0027] As described herein, the present disclosure provides methods of making a suspension of particles comprising at least one therapeutic agent, e.g., a small molecule, a peptide, a protein, an antibody, antibody fragment, antibody-drug conjugate, radionuclide antibody-conjugate, immune-stimulating antibody conjugate, antibody drug conjugate,54248642. vl5726.1016001antibody-oligonucleotide conjugate, or human serum albumin (HSA). In some embodiments, the therapeutic agent is a therapeutic biologic. In some embodiments, the therapeutic biologic is an antibody, antibody fragment, antibody conjugate, or human serum albumin (HSA). In some embodiments one of the therapeutic agents is a hyaluronidase. In some embodiments, a first liquid stream comprises an aqueous feed solution. In some embodiments, the aqueous feed solution comprises at least one therapeutic agent and an aqueous fraction. The generation of particles can be accomplished by generating aqueous droplets comprising a therapeutic biologic and an aqueous fraction. In some embodiments, the aqueous fraction includes an aqueous liquid such as water. The aqueous droplets may be contacted with an organic solvent to dehydrate the aqueous droplets and remove the aqueous fraction by a process (e.g., a continuous process). In certain embodiments, the aqueous droplets are miscible, partially miscible, or immiscible with the organic liquid. In preferred embodiments, the aqueous droplets are immiscible with the organic liquid, / .< ., do not form a uniform solution.

[0028] The aqueous fraction may be removed from the aqueous droplets by contacting the aqueous droplets comprising the therapeutic agent (e.g., therapeutic biologic) with a dehydration liquid (e.g., an organic solvent) by a process in which the aqueous fraction (e.g., water, buffer), but not the therapeutic biologic, is partially soluble or insoluble, thereby leaving behind a particle. In some embodiments, the aqueous droplets are miscible, partially miscible, or immiscible with the dehydration liquid. In preferred embodiments, the aqueous droplets are immiscible with the dehydration liquid.

[0029] The dehydration liquid may dry the aqueous droplets, for example, by removing the aqueous fraction from the aqueous droplets, leaving behind the therapeutic agent as a particle in the organic liquid, in a carrier liquid, on a membrane, or on a filter. In some embodiments, isolation of the particles occurs following removal of the aqueous fraction and / or organic liquid from a fluid channel. In some embodiments, after dehydration, the dehydration liquid may be exchanged with a carrier liquid (e.g., a pharmaceutically acceptable carrier liquid) to maintain the particles in suspension (e.g., with no powder intermediate).

[0030] In certain embodiments, after dehydration, the dehydration liquid may be removed removing the particles from the suspension to form mostly dry particles. The mostly dry particles may be dried by washing the mostly dry particles with the dehydration liquid such as an organic solvent. In some embodiments, the washing step may occur 2 or more times. In certain embodiments, the mostly dry powder may be dried using a fluidized bed dryer alone or in addition to the washing step to form a dry powder comprising at least one therapeutic. The dried particles may form a powder.64248642. vl5726.1016001

[0031] A process of making a suspension of particles comprising a therapeutic agent, as described herein, may significantly alter the structure and morphology of the resulting particles and enhance the stability of the therapeutic agent. Particles of the present disclosure may be used to generate stabilized pharmaceutical compositions, pharmaceutical suspension formulations, pharmaceutical powder formulations (e.g., inhalable powders, injectable powders, and powders that may be reconstituted with a liquid for injection), creams or other topical pastes, nutraceuticals, or cosmetics. The term “pharmaceutical composition,” as used herein, denotes a composition comprising a therapeutic agent (e.g, a therapeutic biologic) in which a therapeutic agent (e.g., a therapeutic biologic) retains, or partially retains, its intended biological activity or functional form, and in which only pharmaceutically acceptable components are included.

[0032] It will be readily understood that the aspects and embodiments, as generally described herein, are examples or exemplary. The following more detailed description of various aspects and embodiments are not intended to limit the scope of the present disclosure, but is merely representative of various aspects and embodiments. Moreover, the methods disclosed herein may be changed by those skilled in the art without departing from the scope of the present disclosure. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in the art to which this disclosure belongs. All publications and patents referred to herein are incorporated by reference.Definitions

[0033] For purposes of the present disclosure, the following definitions will be used unless expressly stated otherwise:

[0034] The terms “a”, “an”, “the” and similar referents used in the context of describing the present disclosure are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. All methods described herein, can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., “such as”) provided herein, is intended merely to better illuminate the present disclosure and does not pose a limitation on the scope of the present disclosure otherwise claimed. No language in the present specification should be construed as indicating any unclaimed element is essential to the practice of the present disclosure.74248642. vl5726.1016001

[0035] The term “about” in relation to a given numerical value, such as for temperature and period of time, is meant to include numerical values within 10% of the specified value.

[0036] Unless otherwise defined, all terms of art, notations and other scientific terminology used herein, are intended to have the meanings commonly understood by those of skill in the art to which this disclosure pertains. In some cases, terms with commonly understood meanings are defined herein, for clarity and / or for ready reference, and the inclusion of such definitions herein, should not necessarily be construed to represent a substantial difference over what is generally understood in the art. The techniques and procedures described or referenced herein, are generally well understood and commonly employed using conventional methodology by those skilled in the art. As appropriate, procedures involving the use of commercially available kits and reagents are generally carried out in accordance with manufacturer defined protocols and / or parameters unless otherwise noted. As used herein, the phrase “and / or” when used in a list of two or more items, means that any one of the listed items can be employed by itself or any combination of two or more of the listed items can be employed. For example, if a composition is described as containing or excluding components A, B, and / or C, the composition can contain or exclude A alone; B alone; C alone; A and B in combination; A and C in combination; B and C in combination; or A, B, and C in combination.

[0037] An organic liquid, organic solvent, dehydration liquid, or carrier liquid may include an alkyl group or alkane. As used herein, a “alkyl” group or “alkane” is a straight chained or branched non-aromatic hydrocarbon which is completely saturated. Typically, a straight chained or branched alkyl group has from 1 to about 20 carbon atoms, preferably from 1 to about 10 unless otherwise defined. Examples of straight chained and branched alkyl groups include methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, tert-butyl, n-pentyl, tert-pentyl, neo-pentyl, iso-pentyl, sec-pentyl, 3-pentyl, sec-iso-pentyl, active-pentyl, hexyl, heptyl, octyl, ethylhexyl, and the like. A Ci-8 straight chained or branched alkyl group is also referred to as a “lower alkyl” group. An alkyl group with two open valences is sometimes referred to as an alkylene group, such as methylene, ethylene, propylene and the like. Moreover, the term “alkyl” (or “lower alkyl”) as used throughout the specification, examples, and claims is intended to include both “unsubstituted alkyls” and “substituted alkyls”, the latter of which refers to alkyl moieties having substituents replacing a hydrogen on one or more carbons of the hydrocarbon backbone. Such substituents, if not otherwise specified, can include, for example, an alkyl, a halogen, a hydroxyl, a carbonyl (such as a carboxyl, and alkoxycarbonyl, a formyl, or an acyl), a thiocarbonyl (such as a thioester, a thioacetate, or a thioformate), an alkoxyl, a phosphoryl, a phosphate, a phosphonate, a phosphinate, an amino,84248642. vl5726.1016001an amido, an amidine, an imine, a cyano, a nitro, an azido, a sulfhydryl, an alkylthio, a sulfate, a sulfonate, a sulfamoyl, a sulfonamido, a sulfonyl, a heterocyclyl, an aralkyl, or an aromatic or heteroaromatic moiety. It will be understood by those skilled in the art that the moieties substituted on the hydrocarbon chain can themselves be substituted, if appropriate. For instance, the substituents of a substituted alkyl may include substituted and unsubstituted forms of amino, azido, imino, amido, phosphoryl (including phosphonate and phosphinate), sulfonyl (including sulfate, sulfonamide, sulfamoyl and sulfonate), and silyl groups, as well as ethers, alkylthios, carbonyls (including ketones, aldehydes, carboxylates, and esters), -CF3, -CN and the like. Example substituted alkyls are described below. Cycloalkyls can be further substituted with alkyls, alkenyls, alkoxys, alkylthios, aminoalkyls, carbonyl-substituted alkyls, -CF3, -CN and the like. In other embodiments, the term “alkyl” can mean “cycloalkyl” which refers to a non-aromatic carbocyclic ring having 3 to 10 carbon ring atoms, which are carbon atoms bound together to form the ring. The ring may be saturated or have one or more carbon-carbon double bonds. Examples of cycloalkyl include, but not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, and cycloheptyl, as well as bridged and caged saturated ring groups such as norbomyl and adamantyl. As described herein, organic solvents and / or dehydration liquids include, but are not limited to aliphatic hydrocarbon solvents, aromatic hydrocarbon solvents, alcohols or alkylalcohols, alkylethers, sulfoxides, alkylketones, alkylacetates, trialkylamines, alkylformates, trialkylamines, or a combination thereof. Aliphatic hydrocarbon solvents can be pentane, hexane, heptane, octane, cyclohexane, and the like or a combination thereof. Aromatic hydrocarbon solvents can be benzene, toluene, and the like or a combination thereof. Alcohols or alkylalcohols include, for example, methanol, ethanol, propanol, butanol, pentanol, hexanol, heptanol, octanol, decanol, amylalcohol, or a combination thereof. Alkylethers include methyl, ethyl, propyl, butyl, and the like, e.g., di ethylether, diisopropylether or a combination thereof. Sulfoxides include dimethyl sulfoxide (DMSO), decylmethyl sulfoxide, tetradecylmethyl sulfoxide, and the like or a combination thereof. The term “alkylketone” refers to a ketone substituted with an alkyl group, e.g., acetone, ethylmethylketone, and the like or a combination thereof. The term “alkylacetate” refers to an acetate substituted with an alkyl group, e.g., ethylacetate, propylacetate (n-propylacetate, iso-propylacetate), butylacetate (n-butyl acetate, isobutylacetate, sec-butylacetate, tert-butylacetate), amylacetate (n-pentylacetate, tertpentylacetate, neo-pentylacetate, iso-pentylacetate, sec-pentylacetate, 3 -pentylacetate, sec-iso-pentylacetate, active-pentylacetate), 2-ethylhexylacetate, and the like or a combination thereof. The term “alkylformate” refers to a formate substituted with an alkyl group, e.g.,94248642. vl5726.1016001methylformate, ethylformate, propylformate, butylformate, and the like or a combination thereof. The term “trialkylamine” refers to an amino group substituted with three alkyl groups, e.g., tri ethylamine.

[0038] As used herein, an “amino acid” or “residue” refers to any naturally or non-naturally occurring amino acid, any amino acid derivative or any amino acid mimic known in the art. Included are the L- as well as the D-forms of the respective amino acids, although the L-forms are usually preferred. In some embodiments, the term relates to any one of the 20 naturally occurring amino acids: glycine (Gly), alanine (Ala), valine (Vai), leucine (Leu), isoleucine (He), proline (Pro), cysteine (Cys), methionine (Met), serine (Ser), threonine (Thr), glutamine (Gin), asparagine (Asn), glutamic acid (Glu), aspartic acid (Asp), lysine (Lys), histidine (His), arginine (Arg), phenylalanine (Phe), tryptophan (Trp), and tyrosine (Tyr) in their L-form. In certain embodiments, the amino acid side-chain may be a side-chain of Gly, Ala, Vai, Leu, He, Met, Cys, Ser, Thr, Trp, Phe, Lys, Arg, His, Tyr, Asn, Gin, Asp, Glu, or Pro.

[0039] “Antigen binding fragments” of antibodies as described herein, comprise only a portion of an intact antibody, generally including an antigen binding site of the intact antibody and thus retaining the ability to bind antigen. Examples of antibody fragments encompassed by the present definition include but are not limited to: (i) the Fab fragment, having VL, CL, VH and CHI domains; (ii) the Fab’ fragment, which is a Fab fragment having one or more cysteine residues at the C-terminus of the CHI domain; (iii) the Fd fragment having VH and CHI domains; (iv) the Fd’ fragment having VH and CHI domains and one or more cysteine residues at the C-terminus of the CHI domain; (v) the Fv fragment having the VL and VH domains of a single arm of an antibody; (vi) the dAb fragment which consists of a VH domain; (vii) isolated CDR regions; (viii) F(ab’)2 fragments, a bivalent fragment including two Fab’ fragments linked by a disulfide bridge at the hinge region; (ix) single chain antibody molecules (e.g. single chain Fv; scFv); (x) “diabodies” with two antigen binding sites, comprising a heavy chain variable domain (VH) connected to a light chain variable domain (VL) in the same polypeptide chain; (xi) “linear antibodies” comprising a pair of tandem Fd, segments (VH-CHl-VH-CHl) which, together with complementary light chain polypeptides, form a pair of antigen binding regions. In some embodiments, an “antigen binding site” generally refers to a molecule that includes at least the hypervariable and framework regions that are required for imparting antigen binding function to a V domain. An antigen binding site may be in the form of an antibody or an antibody fragment, (such as a dAb, Fab, Fd, Fv, F(ab’)2 or scFv) in a method described herein. In some embodiments, an antigen-binding fragment competes with intact antibody, e.g., with the intact antibody from which the fragment was derived, for antigen binding.104248642. vl5726.1016001

[0040] The terms “antibody” and “immunoglobulin” are used interchangeably in the broadest sense and include monoclonal antibodies, polyclonal antibodies, multivalent antibodies and fragments thereof, and multispecific antibodies, regardless of how they are produced (i.e., using immunization or recombinant). Antibodies can be gamma globulin proteins that are found in blood, or other bodily fluids of vertebrates that function in the immune system to bind antigen, hence identifying and / or neutralizing foreign objects. Antibodies can be assigned to different classes or isotypes. There are five classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, having heavy chains designated alpha, delta, epsilon, gamma, and mu, respectively. The gamma class is further divided into subclasses based on the differences in sequences and function, e.g., humans express the following subclasses: IgGl, IgG2, IgG3, IgG4, IgAl, and IgA2. In certain embodiments of the present disclosure, the IgG antibody is a human antibody. The L chain from any vertebrate species may be assigned to one of two clearly distinct types, e.g, kappa and lambda, based on the amino acid sequences of their constant domains.

[0041] The recognized immunoglobulin genes include the kappa, lambda, alpha, gamma, delta, epsilon and mu constant region genes, as well as the myriad immunoglobulin variable region genes. In some embodiments, light chains are classified as either kappa or lambda. In other embodiments, heavy chains are classified as gamma, mu, alpha, delta, or epsilon, which in turn define the immunoglobulin classes, IgG, IgM, IgA, IgD and IgE, respectively. In embodiments of the present disclosure, the antibody is an IgG antibody.

[0042] An example antibody (immunoglobulin) structural unit comprises a tetramer. Each tetramer is composed of two identical pairs of polypeptide chains, each pair having one “light” (about sss25 kD) and one “heavy” chain (about 50-70 kD). The N-terminus of each chain defines a variable region of about 100 to 110 or more amino acids primarily responsible for antigen recognition. The terms “variable light” chain, domain, region and component are used interchangeably, are abbreviated by “VL” or “VL” and refer to the light chain of an antibody or antibody fragment. Similarly, terms “variable heavy” chain, domain, region and component are used interchangeably, are abbreviated by “VH” or “VH” and refer to the heavy chain of an antibody or antibody fragment. Antibodies are generally a heterotetrameric glycoprotein composed of two identical light (L) chains and two identical heavy (H) chains. Each L chain is linked to a H chain by one covalent disulfide bond. The two H chains are linked to each other by one or more disulfide bonds depending on the H chain isotype. Each H and L chain also has regularly spaced intra-chain disulfide bridges. H and L chains define specific Ig domains. In particular, each H chain has at the N-terminus, a variable domain (VH) followed by three 114248642. vl5726.1016001constant domains (CH) for each of the alpha and gamma chains and four CH domains for p and c isotypes. Each L chain has at the N-terminus, a variable domain (VL) followed by a constant domain (CL) at its other end. The VL is aligned with the VH and the CL is aligned with the first constant domain of the heavy chain (CHL). The constant domain includes the Fc portion which comprises the carboxy -terminal portions of both H chains held together by disulfides. The effector functions of antibodies such as ADCC are determined by sequences in the Fc region, which is also the part recognized by Fc receptors (FcR) found on certain types of cells.

[0043] As disclosed herein, the pairing of a VH and VL together form a “variable region” or “variable domain” including the amino-terminal domains of the heavy or light chain of the antibody. The variable domain of the heavy chain may be referred to as “VH”. The variable domain of the light chain may be referred to as “VL”. The V domain contains an “antigen binding site” which affects antigen binding and defines specificity of a particular antibody for its particular antigen. V regions span about 110 amino acid residues and consist of relatively invariant stretches called framework regions (FRs) (generally about 4) of 15-30 amino acids separated by shorter regions of extreme variability called “hypervariable regions” (generally about 3) that are each generally 9-12 amino acids long. The FRs largely adopt a p-sheet configuration and the hypervariable regions form loops connecting, and in some cases forming part of, the p-sheet structure. In certain embodiments, the “hypervariable region” refers to the regions of an antibody variable domain which are hypervariable in sequence and / or form structurally defined loops. Generally, antibodies comprise six hypervariable regions; three in the VH (Hl, H2, H3), and three in the VL (LI, L2, L3). “Framework” or “FR” residues are those variable domain residues other than the hypervariable region residues defined herein.

[0044] The terms “full length antibody”, “intact antibody” and “whole antibody” are used herein, interchangeably, to refer to an antibody in its substantially intact form, not as antibody fragments as defined above. The terms particularly refer to an antibody with heavy chains that contain the Fc region. A full length antibody can be a native sequence antibody or an antibody variant. In certain embodiments, an “intact” or “whole” antibody is one which comprises an antigen-binding site as well as a CL and at least heavy chain constant domains, CHI, CH2 and CH3. The constant domains may be native sequence constant domains (e.g. human native sequence constant domains) or amino acid sequence variants thereof.

[0045] As disclosed herein, “whole antibody fragments including a variable domain” include Fab, Fab’, F(ab’)2, and Fv fragments; diabodies; linear antibodies, single-chain antibody molecules; and multispecific antibodies formed from antibody fragments. The “Fab fragment” consists of an entire L chain along with the variable region domain of the H chain 124248642. vl5726.1016001(VH), and the first constant domain of one heavy chain (CHI). Each Fab fragment is monovalent with respect to antigen binding, z.e., it has a single antigen-binding site. A “Fab’ fragment” differs from Fab fragments by having additional few residues at the carboxy terminus of the CHI domain including one or more cysteines from the antibody hinge region. Fab’-SH is the designation herein for Fab’ in which the cysteine residue(s) of the constant domains bear a free thiol group. A “F(ab’)2 fragment” roughly corresponds to two disulfide linked Fab fragments having divalent antigen-binding activity and is still capable of crosslinking antigen. An “Fv” is the minimum antibody fragment which contains a complete antigen-recognition and binding site. This fragment consists of a dimer of one heavy and one light chain variable region domain in tight, non-covalent association. In a single-chain Fv (scFv) species, one heavy and one light chain variable domain can be covalently linked by a flexible peptide linker such that the light and heavy chains can associate in a “dimeric” structure analogous to that in a two-chain Fv species. From the folding of these two domains emanate six hypervariable loops (3 loops each from the H and L chain) that contribute the amino acid residues for antigen binding and confer antigen binding specificity to the antibody. “Singlechain Fv” also abbreviated as “sFv” or “scFv” are antibody fragments that comprise the VH and VL antibody domains connected to form a single polypeptide chain. In embodiments, the scFv polypeptide further comprises a polypeptide linker between the VH and VL domains which enables the scFv to form the desired structure for antigen binding. In some embodiments, a “single variable domain” is half of an Fv (comprising only three CDRs specific for an antigen) that has the ability to recognize and bind antigen, although at a lower affinity than the entire binding site.

[0046] In some embodiments, “diabodies” refer to antibody fragments with two antigenbinding sites, which fragments comprise a heavy-chain variable domain (VH) connected to a light-chain variable domain (VL) in the same polypeptide chain (VH-VL). The small antibody fragments are prepared by constructing sFv fragments with short linkers (about 5-10 residues) between the VH and VL domains such that inter-chain but not intra-chain pairing of the V domains is achieved, resulting in a bivalent fragment, i. e. , fragment having two antigen-binding sites. In other embodiments, diabodies may be bivalent or bispecific. In certain embodiments, bispecific diabodies are heterodimers of two “crossover” sFv fragments in which the VH and VL domains of the two antibodies are present on different polypeptide chains. Triabodies and tetrabodies are also generally known in the art.

[0047] The term “fragment” refers to a part or portion of an antibody or antibody chain comprising fewer amino acid residues than an intact or complete antibody or antibody chain.134248642. vl5726.1016001As used herein, the term “fragment” of an antibody includes Fc fragments and antigen-binding fragments of antibodies, for example, an antibody light chain variable domain (VL), an antibody heavy chain variable domain (VH), a single chain antibody (scFv), a F(ab’)2 fragment, a Fab fragment, an Fd fragment, an Fv fragment, a single domain antibody fragment (DAb), a one-armed (monovalent) antibody, or any antigen-binding molecule formed by combination, assembly or conjugation of such antigen binding fragments. In particular embodiments, the fragment is an Fc fragment.

[0048] In certain embodiments, the term “single-chain Fv” or “scFv” or “single chain” antibody can refer to antibody fragments comprising the VH and VL domains of antibody, wherein these domains are present in a single polypeptide chain. Generally, the Fv polypeptide further comprises a polypeptide linker between the VH and VL domains which enables the sFv to form the desired structure for antigen binding. For a review of sFv, see Pluckthun, THE PHARMACOLOGY OF MONOCLONAL ANTIBODIES, vol. 113, Rosenburg and Moore eds. Springer-Verlag, New York, pp. 269-315 (1994).

[0049] As used herein, the term “monoclonal antibody” refers to an antibody obtained from a population of substantially homogeneous antibodies, / .< ., the individual antibodies comprising the population are identical except for possible naturally occurring variations that may be present in minor amounts. Variations may be due to degradation or post-translational modifications. Monoclonal antibodies (mAbs) are highly specific, being directed against a single antigenic site or determinant on the antigen. In addition to their specificity, the monoclonal antibodies are advantageous in that they may be synthesized uncontaminated by other antibodies. Monoclonal antibodies may be prepared by the hybridoma methodology. The monoclonal antibodies may also be isolated from phage antibody libraries using molecular engineering techniques. Monoclonal antibodies of the present disclosure may be generated by recombinant DNA methods, and are sometimes referred to as “recombinant antibodies” or “recombinant monoclonal antibodies” as described herein. In some embodiments, a monoclonal antibody is a single species of antibody wherein every antibody molecule recognizes the same epitope because all antibody producing cells are derived from a single B-lymphocyte cell line. Methods for generating monoclonal antibodies (mAbs) generally begin along the same lines as those for preparing polyclonal antibodies. In other embodiments, rodents such as mice and rats are used in generating monoclonal antibodies. In certain embodiments, rabbit, sheep, or frog cells are used in generating monoclonal antibodies. The use of rat cells is well known and may provide certain advantages. Mice cells (e.g., BALB / c mice) are routinely used and generally give a high percentage of stable fusions. Similarly,144248642. vl5726.1016001Chinese hamster ovary cells may be used as known in the art. In still other embodiments of the present disclosure, the antibody is a monoclonal antibody. In embodiments of the present disclosure, the IgG antibody is monoclonal.

[0050] In other embodiments, recombinant antibody fragments may be isolated from phage antibody libraries using techniques well known in the art. See, for example, Clackson et al., 1991, Nature 352: 624-628; Marks et al., 1991, J. Mol. Biol. 222: 581-597. Recombinant antibody fragments may be derived from large phage antibody libraries generated by recombination in bacteria (Sblattero and Bradbury, 2000, Nature Biotechnology 18:75-80; and as described herein). Polynucleotides encoding the VH and VL components of antibody fragments (i.e., scFv) may be used to generate recombinant full length immunoglobulins using methods known in the art (see, for example, Persic et al., 1997, Gene 187: 9-18).

[0051] An “isolated antibody” is one that has been identified and separated and / or recovered from a component of its pre-existing environment. Contaminant components are materials that would interfere with therapeutic uses for the antibody, and may include enzymes, hormones, and other proteinaceous or nonproteinaceous solutes.

[0052] As used herein, a “human antibody” refers to an antibody that possesses an amino acid sequence that corresponds to that of an antibody produced by a human. Human antibodies can be produced using various techniques known in the art, including phage-display libraries. Human antibodies can be prepared by administering the antigen to a transgenic animal that has been modified to produce such antibodies in response to antigenic challenge, but whose endogenous loci has been disabled. “Humanized” forms of non-human (e.g., rodent) antibodies are chimeric antibodies that contain minimal sequence derived from the non-human antibody. For the most part, humanized antibodies are human immunoglobulins (recipient antibody) in which residues from a hypervariable region of the recipient are replaced by residues from a hypervariable region of a non-human species (donor antibody) such as mouse, rat, rabbit or non-human primate having the desired antibody specificity, affinity, and capability. In some instances, framework region (FR) residues of the human immunoglobulin are replaced by corresponding non-human residues. Furthermore, humanized antibodies may comprise residues that are not found in the recipient antibody or in the donor antibody. These modifications are made to further refine antibody performance. In general, the humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin and all or substantially all of the FRs are those of a human immunoglobulin154248642. vl5726.1016001sequence. The humanized antibody optionally also will comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin.

[0053] An “affinity matured” antibody is one with one or more alterations in one or more hypervariable region thereof that result in an improvement in the affinity of the antibody for antigen, compared to a parent antibody that does not possess those alterations. In some embodiments, affinity matured antibodies can have micromolar affinities for the target antigen. In other embodiments, affinity matured antibodies can have nanomolar or even picomolar affinities for the target antigen. Affinity matured antibodies are produced by procedures known in the art.

[0054] The term “aseptic” as used herein, with respect to all or part of a process (e.g., a step), refers to a process or part of a process that is designed to maintain sterility and prevent or minimize microbial contamination. For example, an aseptic process of forming particles or of making a particle suspension can begin with a sterilized solution (e.g., a sterilized aqueous feed and organic liquid) and conclude with particles or a particle suspension that is / are sterile. Such aseptic processes can be batch, continuous, or semi-continuous. In such aseptic processes, sterility can be obtained, for example, by using sterilized equipment and by processing within controlled environmental conditions that have been demonstrated to produce sterile material. In one embodiment, an aseptic process is provided of forming particles or of making a particle suspension, in which a dehydration liquid (e.g., an organic solvent) may be exchanged with a carrier liquid (e.g., a pharmaceutically acceptable carrier liquid) to maintain the particles in suspension with no powder intermediate.

[0055] A “blocking” antibody or an “antagonist” antibody is one that inhibits or reduces biological activity of the antigen it binds. In some embodiments, blocking antibodies or antagonist antibodies substantially or completely inhibit the biological activity of the antigen. An “agonist antibody”, as used herein, is an antibody, which mimics at least one of the functional activities of a polypeptide of interest.

[0056] “Binding affinity” generally refers to the strength of the sum total of noncovalent interactions between a single binding site of a molecule, e.g., an antibody, and its binding partner, e.g., an antigen. Unless indicated otherwise, as used herein, “binding affinity” refers to intrinsic binding affinity, which reflects a 1 : 1 interaction between members of a binding pair, e.g., antibody and antigen. The affinity of a molecule X for its partner Y can generally be represented by the dissociation constant (Kd). Affinity can be measured by common methods known in the art, including those described herein. Low-affinity antibodies generally bind antigen slowly and tend to dissociate readily, whereas high-affinity antibodies generally bind 164248642. vl5726.1016001antigen faster and tend to remain bound longer. A variety of methods of measuring binding affinity are known in the art, any of which can be used for purposes of the present disclosure. “Epitope” generally refers to that part of an antigen that is bound by the antigen binding site of an antibody. In some embodiments, an epitope may be “linear” in the sense that the hypervariable loops of the antibody CDRs that form the antigen binding site bind to a sequence of amino acids as in a primary protein structure. In other embodiments, the epitope is a “conformational epitope”, i.e. one in which the hypervariable loops of the CDRs bind to residues as they are presented in the tertiary or quaternary protein structure.

[0057] As used herein, except where the context requires otherwise, the term “comprise” and variations of the term, such as “comprising”, “comprises” and “comprised”, are not intended to exclude further additives, components, integers or steps. The terms “including” and “comprising” may be used interchangeably. As used herein, the phrases “selected from the group consisting of’, “chosen from”, and the like, include mixtures of the specified materials. Where a numerical limit or range is stated herein, the endpoints are included. Also, all values and subranges within a numerical limit or range are specifically included as if explicitly written herein. References to an element in the singular is not intended to mean “one and only one” unless specifically stated, but rather “one or more”. Unless specifically stated otherwise, terms such as “some” refer to one or more, and singular terms such as “a”, “an” and “the” refer to one or more.

[0058] As used herein, “continuous” means, with respect to all or a portion of a process (e.g., one or more steps), that all or a portion of the process proceeds in succession and without interruption for a period of time (e.g., about 1 to about 60 seconds, about 1 minute to about 60 minutes, about 1 hour to about 24 hours, about 1 day to about 7 days, and about 1 week to about 4 weeks). As used herein, the phrase “semi-continuous” means, with respect to all or a portion of a process (e.g., one or more steps) which may be interrupted or stopped for a period of time. Embodiments of the process disclosed herein may be continuous or semi-continuous. In some embodiments of the process disclosed herein, each step of the process may be continuous or semi-continuous.

[0059] Oligopeptides described herein, are typically comprised of about two to about forty amino acid residues. Oligopeptides include dipeptides (two amino acids), tripeptides (three amino acids), tetrapeptides (four amino acids), pentapeptides (five amino acids), hexapeptides (six amino acids), heptapeptides (seven amino acids), octapeptides (eight amino acids), nonapeptides (nine amino acids), decapeptides (ten amino acids), undecapeptides (eleven amino acids), dodecapeptides (twelve amino acids), icosapeptides (twenty amino acids),174248642. vl5726.1016001tricontapeptides (thirty amino acids), tetracontapeptides (forty amino acids), and the like. Oligopeptides may also be classified according to molecular structure: aeruginosins, cyanopeptolins, microcystins, microviridins, microginins, anabaenopeptins and cyclamides, and the like. Homo-oligopeptides are oligopeptides comprising the same amino acid. In embodiments, homo-oligopeptides comprise 10 amino acid poly-valine, poly-alanine, and poly-glycine hexamers.

[0060] The meaning of the term “peptides” are defined as small proteins of two or more amino acids linked by the carboxyl group of one to the amino group of another. Accordingly, at its basic level, peptide synthesis of whatever type comprises the repeated steps of adding amino acid or peptide molecules to one another or to an existing peptide chain. The term “peptide” generally has from about 2 to about 100 amino acids, whereas a polypeptide or protein has about 100 or more amino acids, up to a full length sequence which may be translated from a gene. Additionally, as used herein, a peptide can be a subsequence or a portion of a polypeptide or protein. In certain embodiments, the peptide consists of 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100 amino acid residues. In embodiments, the peptide is from about 30 to about 100 amino acids in length. In some embodiments, the peptide is from about 40 to about 100 amino acids in length.

[0061] As used herein, the term “pharmaceutically acceptable” refers to compositions that are physiologically tolerable and do not typically produce an allergic or similar untoward reaction when administered to a subject, preferably a human subject.

[0062] As used herein, the term “prodrug” is intended to encompass therapeutic agents which, under physiologic conditions, are converted into the therapeutically active agents of the present disclosure. A common method for making a prodrug is to include one or more selected moieties which are hydrolyzed under physiologic conditions to reveal the desired molecule. In other embodiments, the prodrug is converted by an enzymatic activity of the host animal. For example, esters or carbonates (e.g., esters or carbonates of alcohols or carboxylic acids) are prodrugs contemplated by the present disclosure. In certain embodiments, some or all of the molecules in a composition represented above can be replaced with the corresponding suitable prodrug, e.g., wherein a hydroxyl in the parent molecule is presented as an ester or a carbonate or carboxylic acid present in the parent therapeutic agent is presented as an ester.184248642. vl5726.1016001

[0063] The meaning of the term “protein” is defined as a linear polymer built from about 20 different amino acids. The type and the sequence of amino acids in a protein are specified by the DNA that produces them. In certain embodiments, the sequences can be natural and unnatural. The sequence of amino acids determines the overall structure and function of a protein. In some embodiments, proteins can contain 50 or more residues. In embodiments, proteins can contain greater than about 101 residues in length. A protein's net charge can be determined by two factors: 1) the total count of acidic amino acids vs. basic amino acids; and 2) the specific solvent pH surroundings, which expose positive or negative residues. As used herein, “net positively or net negatively charged proteins” are proteins that, under nondenaturing pH surroundings, have a net positive or net negative electric charge. In general, those skilled in the art will recognize that all proteins may be considered “net negatively charged proteins”, regardless of their amino acid composition, depending on their pH (such as if the pH is less than 9.5) and / or solvent surroundings. Proteins or peptides are preferably selected from any type of enzyme or antibodies or fragments thereof. Proteins or peptides may serve as a structural material (e.g. keratin), as enzymes, as hormones, as transporters (e.g. hemoglobin), as antibodies, or as regulators of gene expression. Proteins or peptides are required for the structure, function, and regulation of cells, tissues, and organs.

[0064] The term “substantially” as used herein, refers to a majority of, or mostly, as in at least about 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%, 99.99%, or at least about 99.999% or more.

[0065] The terms “particle” or “particles” or “microparticle” or “microparticles” are used herein, interchangeably in the broadest sense, refers to a discrete body or bodies. The particles described herein, are circular, spheroidal and of controlled poly dispersity across a population of particles with a characteristic size from sub-micrometers to tens of micrometers, in contrast to, e.g, a porous monolithic “cake”, which is typically produced during conventional lyophilization. The poly dispersity of the population of particles may be measured by taking a weighted average or weighted volume average depending on the characteristic of the population or a portion of the population. This morphology allows for a flowable powder (as described by low Hausner ratios) without post-processing. Additionally, the controlled smoothness, circularity and spherical shape of the particles increases the volume available for at least one therapeutic forming at least a portion of the particle(s) thereby increasing an achievable density of therapeutic across a portion of the population of particles. In some embodiments, the term “particle” refers to a quantity of a therapeutic agent or therapeutic agents which is either in a state of matter that is substantially solid as compared to a liquid 194248642. vl5726.1016001droplet or in a gel form. The term “proto-particle” refers to a stage of particle formation in which one or more of the components comprising the particle are in an at least a partial state of desiccation. For example, methods described herein may include forming particles by, at least in part, dehydrating an aqueous fraction of an aqueous droplet. The dehydrating aqueous droplet transitioning to a particle as described herein may be referred to at each stage of dehydration and throughout the dehydration process as a “proto-particle” as contemplated by this disclosure. In some embodiments, the particles or proto-particles may be in a suspension. The total liquid content of the proto-particle is less than that of the aqueous droplet and greater than that of the formed particle. Similarly, the volume-weighted average density of a therapeutic in the aqueous droplet is typically less than that of the formed particle. In certain embodiments of the present disclosure, proto-particles are formed in a step of a method of the present disclosure. The term “encapsulant” refers to a substance that can be dried or gelled around a particle core to form a shell.

[0066] As disclosed herein, a therapeutic agent, also known as a biologic medical product, or biopharmaceutical, is any pharmaceutical drug product manufactured in, extracted from, or semi synthesized from biological sources. Therapeutic agents can include a wide range of products such as vaccines, blood and blood components, allergenics, somatic cells, gene therapy, tissues, and recombinant therapeutic proteins. In some embodiments, biologies may include sugars, proteins, or nucleic acids or complex combinations of these substances, or may be living entities such as cells and tissues. Biologies can be isolated from a variety of natural sources, e.g. , a human, animal, microorganism, or plant and may be produced by biotechnology methods or other technologies. Gene-based and cellular biologies, for example, are often used to treat a variety of medical conditions for which no other treatments are available. In certain embodiments of the present disclosure, a therapeutic agent is an antibody or fragment thereof, or human serum albumin (HSA). In certain embodiments, the antibody is a human antibody, e.g., human IgG, or a monoclonal antibody (mAb). In certain other embodiments, the antibody is Rituximab or Trastuzumab. In still other embodiments, the antibody is a fragment of an antibody.

[0067] In some embodiments of the foregoing methods, the therapeutic agent is an antibody. Examples of antibodies contemplated by this disclosure are disclosed in U.S. Patent Publication No. 2024 / 0270864, the entirety of which is incorporated by reference herein. In other embodiments, the targeting moiety is an antibody from an intact polyclonal antibody, an intact monoclonal antibody, an antibody fragment, a single chain Fv (scFv) mutant, a multispecific antibody, a bispecific antibody, a chimeric antibody, a humanized antibody, a 204248642. vl5726.1016001human antibody, a fusion protein comprising an antigenic determinant portion of an antibody, or other modified immunoglobulin molecules comprising antigen recognition sites.

[0068] In some embodiments, the therapeutic agent is an immunotherapy. In other embodiments, the immunotherapy is an anti-CD20 antibody. In certain embodiments, the anti-CD20 antibody is rituximab. In certain other embodiments, the therapeutic agent is an anti-CD20 antibody. As described herein, any antibody capable of binding the CD20 antigen may be used in the methods of the instant disclosure. Antibodies which bind the CD20 antigen include, for example: C2B8 (rituximab; RITUXAN®) (U.S. Pat. No. 5,736,137, expressly incorporated herein by reference); the yttrium-

[0090] -labeled 2138 murine antibody designated Y2B8 (U.S. Pat. No. 5,736,137, expressly incorporated herein by reference); murine IgG2a 131 optionally labeled with 131 1 to generate the 131 1-B1 antibody (BEXXAR®) (U.S. Pat. No. 5,595,721, expressly incorporated herein by reference); murine monoclonal antibody 1F5 (Press et al. Blood 69(2): 584-591 (1987)); chimeric 2H7 antibody (U.S. Pat. No. 5,677,180 expressly incorporated herein by reference); and monoclonal antibodies L27, G28-2, 93-1 133, B— Cl or NU— B2 available from the International Leukocyte Typing Workshop (Valentine et al., In: Leukocyte Typinglll (McMichael, Ed., p. 440, Oxford University Press (1987)).

[0069] In certain embodiments of the present disclosure, the anti-CD20 antibody is rituximab. Rituximab is a genetically engineered chimeric murine / human monoclonal antibody. Rituximab is an IgG, kappa immunoglobulin containing murine light and heavy chain variable region sequences and human constant region sequences. Rituximab has a binding affinity for the CD20 antigen of approximately 8.0 nM and is commercially available, e.g., from Genentech (South San Francisco, Calif.).

[0070] In some embodiments, the therapeutic agent is an immunotherapeutic. Examples of immunotherapeutic agents and immunotherapies contemplated by this disclosure are disclosed in U.S. Patent Publication No. 2024 / 0270864, the entirety of which is incorporated by reference herein.

[0071] The therapeutic agent in the particles may have an activity per unit of about 0.5 to about 1.0, about 0.75 to about 1.0 activity per unit, or about 0.9 to about 1.0 activity per unit. Activity is measured relative to the same therapeutic agent prior to particle formation. In embodiments, the therapeutic agent has an activity per unit of about 0.5 to about 1.0. The term “activity” refers to the ratio of a functional or structural aspect of a therapeutic agent, e.g., an antibody, antibody fragment, or human serum albumin (HSA), at two points in time. The denominator of the ratio corresponds to a measure of the functional or structural aspect of the therapeutic agent in the feed solution, immediately in advance of droplet formation. The 214248642. vl5726.1016001numerator of the ratio corresponds to the same measure of a functional or structural aspect of the therapeutic agent at a later point in time, e.g., immediately after particle formation.

[0072] It is understood that the specific order or hierarchy of steps in the methods or processes disclosed is an illustration of example approaches. Based upon design preferences, it is understood that the specific order or hierarchy of steps in the methods or processes may be rearranged. Some of the steps may be performed simultaneously. The accompanying method claims present elements of the various steps in a sample order, and are not meant to be limited to a specific hierarchy or order presented. A phrase such as “embodiment” does not imply that such embodiment applies to all configurations of the subject technology. A disclosure relating to an embodiment may apply to all embodiments, or one or more embodiments. A phrase such as an embodiment may refer to one or more embodiments and vice-versa.Methods of the present disclosure

[0073] The methods described herein, are generally provided for making a suspension of particles comprising a therapeutic agent as described herein, the method comprising: a) providing a first liquid stream comprising an aqueous feed solution, the feed solution comprising a therapeutic agent; b) providing a second liquid stream comprising an organic liquid; c) dispersing the first liquid stream into the second liquid stream at a junction; d) applying turbulent flow conditions to a fluid channel connected to the junction, the fluid channel containing the first liquid stream and the second liquid stream, to form aqueous droplets comprising the therapeutic agent; e) dehydrating an aqueous fraction of the aqueous droplets, the dehydrating comprising contacting the aqueous droplets with a dehydration liquid to form particles comprising the therapeutic agent; f) removing the dehydration liquid; and g) suspending the particles in a pharmaceutically acceptable carrier liquid to form a suspension, wherein the suspension has a water content of less than about 20% (w / v). As disclosed herein, the therapeutic agent is a therapeutic biologic (e.g., an antibody or antibody fragment). A therapeutic agent’s activity or potency in a portion of the particles may be measured compared to therapeutic agent’s activity or potency in the aqueous feed solution. In certain embodiments, the therapeutic agent has an activity per unit of about 0.5 to about 1.0. In embodiments, the therapeutic agent in a portion of the particles has an activity per unit of about 0.8 to about 1.0. In some embodiments, therapeutic agent in a portion of the particles has about 50% to about 100% potency compared to therapeutic agent in the aqueous feed solution. In certain embodiments, therapeutic agent in a portion of the particles has about 80% to about 100% potency compared to therapeutic agent in the aqueous feed solution. In some embodiments, the224248642. vl5726.1016001second fluid stream is under turbulent flow conditions. In certain embodiments, the first fluid stream is under laminar flow conditions. The activity or potency may be measured using a binding or cell based assay as known in the art.

[0074] In some embodiments, prior to a), the aqueous feed solution is filtered using a process that comprises applying diafiltration or tangential flow filtration and concentration to the aqueous feed solution. In some embodiments, at least one excipient in the aqueous feed solution may be exchanged and replaced with a different excipient. The aqueous feed solution may include an initial excipient profile. In some embodiments, the initial excipient profile of the aqueous feed solution may be added or replaced with a second excipient profile prior to a).

[0075] In some embodiments, step a) includes performing tangential flow filtration. In some embodiments, the tangential flow filtration is performed using a recirculation flow rate of from about 100 mL / min to about 5000 mL / min (e.g., about 100 mL / min to about 4500 mL / min, about 100 mL / min to about 4000 mL / min, about 100 mL / min to about 3500 mL / min, about 100 mL / min to about 3000 mL / min, about 100 mL / min to about 2500 mL / min, about 100 mL / min to about 2000 mL / min, about 500 mL / min to about 2000 mL / min, about 1000 mL / min to about 2000 mL / min, etc.). In some embodiments, the recirculation flow rate is from about 1000 mL / min to about 2000 mL / min.

[0076] In some embodiments, cross-flow filtration is performed using a transmembrane pressure of at least 1 psi, preferably from about 1 psi to about 50 psi (e.g., about 1 psi to about 40 psi, about 1 psi to about 30 psi, about 1 psi to about 20 psi, about 5 psi to about 20 psi, etc.). In some embodiments, the cross-flow filtration is performed using a transmembrane pressure of from about 10 psi to about 15 psi.

[0077] In some embodiments, cross-flow filtration is performed using a membrane loading of at least 100 g / m2, preferably from about 100 to about 1000 g / m2(e.g., about 100 to about 1000 g / m2, about 200 to about 1000 g / m2, about 300 to about 1000 g / m2, about 400 to about 1000 g / m2, about 500 to about 1000 g / m2, about 500 to about 900 g / m2, about 500 to about 800 g / m2, etc.). In some embodiments, the cross-flow filtration is performed using a membrane loading of from about 500 to about 1000 g / m2. In some embodiments, the cross-flow filtration is performed using a membrane loading of from about 500 to about 750 g / m2.

[0078] In some embodiments, cross-flow filtration is performed using a flow rate (e.g., feed flow rate) of from about 100 mL / min to about 5000 mL / min (e.g., about 100 mL / min to about 4500 mL / min, about 100 mL / min to about 4000 mL / min, about 100 mL / min to about 3500 mL / min, about 100 mL / min to about 3000 mL / min, about 100 mL / min to about 2500 mL / min, about 100 mL / min to about 2000 mL / min, about 500 mL / min to about 2000 mL / min,234248642. vl5726.1016001about 1000 mL / min to about 2000 mL / min, etc.). In some embodiments, the feed flow rate is from about 100 mL / min to about 5000 mL / min. In some embodiments, the feed flow rate is from about 100 mL / min to about 2000 mL / min.

[0079] In some embodiments, prior to a), the aqueous feed solution is warmed to a temperature of at least 10°C, 15°C, or 20°C, preferably from about 20°C to about 25°C for a defined duration (e.g., for at least about an hour, at least about two hours, at least about three hours, etc.). In some embodiments, the temperature is from about 15-30°C.

[0080] In some embodiments disclosed herein are methods of making a suspension of particles comprising at least one therapeutic agent, the methods comprising:a) providing a first liquid stream comprising an aqueous feed solution, the feed solution comprising the therapeutic agent;b) providing a second liquid stream comprising an organic liquid;c) dispersing the first liquid stream into the second liquid stream at a first junction;d) applying turbulent flow conditions to a fluid channel connected to the first junction and a second junction, the fluid channel containing the first liquid stream and the second liquid stream, to form aqueous droplets comprising the therapeutic agent;e) dehydrating an aqueous fraction of the aqueous droplets, the dehydrating comprising contacting the droplets with a dehydration liquid to form particles comprising the therapeutic agent; andf) suspending the particles in a carrier liquid to form the suspension of particles aseptically.

[0081] In some embodiments, the organic liquid is filtered using a filter having a pore size of about 0.2 pm. In other embodiments, the organic liquid is filtered using a filter having a pore size of at least about 0.1 pm (e.g., at least about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 10, 15, 25, 50 pm).

[0082] In other embodiments disclosed herein are methods of adjusting the water content of a plurality of particles in a suspension, the method comprising performing sparging on the suspension, wherein the suspension comprises the particles suspended in an organic carrier liquid, and wherein the plurality of the particles comprise a therapeutic agent.

[0083] In some embodiments disclosed herein are methods of forming a suspension of particles comprising a therapeutic agent, the methods comprising:244248642. vl5726.1016001a) providing a first liquid stream comprising an aqueous feed solution, the feed solution comprising the therapeutic agent;b) providing a second liquid stream comprising an organic liquid;c) dispersing the first liquid stream into the second liquid stream at a first junction;d) applying turbulent flow conditions to a fluid channel connected to the first junction and a second junction, the fluid channel containing the first liquid stream and the second liquid stream, to form aqueous droplets comprising the therapeutic agent;e) dehydrating an aqueous fraction of the aqueous droplets, the dehydrating comprising contacting the droplets with a dehydration liquid to form particles comprising the therapeutic agent; andf) collecting the particles aseptically.

[0084] In other embodiments disclosed herein are methods of forming particles comprising a therapeutic agent, the methods comprising:a) providing a first liquid stream comprising an aqueous feed solution comprising the therapeutic agent;b) providing a second liquid stream comprising an organic liquid;c) dispersing the first liquid stream into the second liquid stream at or after a first junction;d) applying turbulent flow conditions to a fluid channel connected to the junction, the fluid channel containing the first liquid stream and the second liquid stream, to form aqueous droplets comprising the therapeutic agent; e) dehydrating an aqueous fraction of the aqueous droplets, the dehydrating comprising contacting the droplets with a dehydration liquid at a second junction to form particles comprising the therapeutic agent, wherein the Peclet number of the combination of the first liquid stream, the second liquid stream and the dehydration liquid determines the morphology of the particles; and f) collecting the particles, thereby forming particles comprising the therapeutic agent, wherein the particles have a circularity of from about 0.80 to about 1.00.

[0085] In some embodiments, a junction (e.g., a junction for dispersing a first liquid stream into a second liquid stream) is provided. In certain embodiments, the junction includes a stream inlet with an internal diameter greater than about 0.1 mm (e.g., greater than about 0.1, 0.2, 0.3,254248642. vl5726.10160010.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 3.5, 4.0, 4.5, 5, etc.). Examples of junctions may include, but are not limited to, a fitting, a Tee-mixer, a Y-mixer, or a static mixer. The junction may include an interface between a first fluid channel and a second fluid channel. The junction may prevent backflow such as by use of at least one check-valve. The first fluid channel and / or the second fluid channel may include a check-valve. The first fluid channel and the second fluid channel may include the same internal diameter. The first fluid channel may include a larger internal diameter than the second fluid channel. The first fluid channel may include a smaller internal diameter than the second fluid channel. In some embodiments, the junction may utilize the first fluid channel and the second fluid channel as inlet channels. In certain embodiments, the junction may include a third fluid channel as an outlet.

[0086] In some embodiments, liquid streams are combined at or within a mixing apparatus having movable parts (e.g., a rotor-stator mixer, a static mixer, or an extruder). In some embodiments, the methods described herein comprise the use of more than one junction. By way of example, an embodiment of the present disclosure may include dispersing a first liquid stream into a second liquid stream at a first junction and applying high-shear forces at a second junction in a fluid channel. By way of further example, an embodiment of the present disclosure may include dispersing a first liquid stream into a second liquid stream at a first junction wherein the fluid channel at the first junction includes highly turbulent flow conditions and applying high-shear forces at a second junction in a fluid channel. In some embodiments, the first liquid stream may be dispersed into a second liquid stream at a first junction, wherein the second liquid stream is under turbulent flow conditions, and applying highly turbulent flow conditions at a second junction. In some embodiments, flow conditions within a fluid channel are highly turbulent. Additional liquids such as an organic solvent or a dehydration liquid may be introduced at any of the one or more junctions. In certain embodiments, the methods disclosed herein do not employ the use of a mixing apparatus having movable parts. In some embodiments, the methods disclosed herein do not use a microfluidic device, static mixer, or homogenizer.

[0087] In some embodiments, turbulent flow conditions are applied using a rotor stator mixer, Tee-mixer, or static mixer. In some embodiments, turbulent flow conditions are applied at a first junction or a second junction. In certain embodiments, the turbulent flow conditions are applied within a fluid channel or fluid vessel. In some embodiments, the first junction comprises a rotor stator mixer, Tee-mixer, or static mixer. In some embodiments, the second junction comprises a rotor stator mixer, Tee-mixer, or static mixer.264248642. vl5726.1016001

[0088] Laminar flow conditions, as known in the art, form when the Reynolds number is less than 2,000. Bird, R. Byron et al., Phenomena Second Edition (2002), In some embodiments, turbulent flow conditions form when the Reynolds number is 2,100 or greater. In some embodiments, highly turbulent conditions form at a Reynolds number of 25,000 or greater. In certain embodiments, the Reynolds number within the junction, fluid channel, or fluid vessel is between about 2,100 and about 100,000 (e.g., between about 2,100-25,000, about 25,000-50,000, about 50,000-75,000, about 75,000-100,000, about 25,000-30,000, about 30,000-40,000, about 40,000-50,000, about 50,000-60,000, about 60,000-70,000, about 70,000-80,000, about 80,000-90,000, about 90,000-100,000, about 2,100-50,000, about 50,000-80,000, about 80,000-100,000).

[0089] In some embodiments, a rotor-stator mixer may be used to combine the aqueous liquid and the organic liquid. In certain embodiments, the rotor-stator mixer may replace the junction. In certain other embodiments, the rotor stator mixer may be fluidly connected to the junction or at the junction where the first fluid channel and the second fluid channel meet. In some embodiments, the rotor-stator mixer may be fluidly connected in-line with the fluid channel. In certain embodiments, the rotor-stator mixer may be a vessel or continuous mixing tank for forming aqueous droplets in an organic liquid from an aqueous liquid which is immiscible in the organic liquid. In some embodiments, the rotor stator mixer may include a vessel containing the aqueous fluid, the organic liquid, or any combination thereof. The rotor stator mixer may apply a shearing force to the liquid(s) to form aqueous droplets of the aqueous fluid in the organic liquid. The methods and systems disclosed herein may utilize any rotorstator mixer known in the art without departing from the scope of the present disclosure.

[0090] In some embodiments, a rotating mixer may float, through magnetic repulsion or other techniques known in the art, within a fluid channel or vessel to apply shear forces on at least one liquid stream in the fluid channel or vessel. The rotating mixer may be operated by manipulating a magnetic field around the rotating mixer to cause rotation. The rotating mixer may be located in a junction in the fluid channel, before a junction, after a junction, between junctions in the fluid channel, or in place of a junction. As a result, moving parts may be introduced into the methods and systems disclosed herein without the introduction of SVPs or contaminants.

[0091] In some embodiments, a first liquid stream and a second liquid stream are combined at a junction having an internal diameter of from about 0.1 mm to about 30 mm (e.g., about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30 mm). In some embodiments, the junction includes a first stream inlet and a second stream inlet. The first 274248642. vl5726.1016001stream inlet and the second stream inlet may be sized with reference to each other such that the first stream inlet is approximately the same size as the second stream inlet (1 : 1), the first stream inlet is approximately twice the size of the second stream inlet (2: 1), or the first stream inlet is approximately half the size of the second stream inlet (1:2). In some embodiments the ratio of the first stream inlet to the second stream inlet may be from about 1:1 to about 1:300 (e.g., about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, 1:11, about 1:12, about 1:13, about 1:14, about 1:15, about 1:16, about 1:17, about 1:18, about 1:19, about 1:20, about 1:21, about 1:22, about 1:23, about 1:24, about 1:25, about 1:26, about 1:27, about 1:28, about 1:29, about 1:30, about 1:50, about 1:100, about 1:150, about 1 :200, about 1 :250, about 1 :300). In some embodiments the ratio of the first stream inlet to the second stream inlet may be from about 1:1 to about 300:1 (e.g., about 1:1, about 2:1, about 3:1, about 4:1, about 5:1, about 6:1, about 7:1, about 8:1, about 9:1, about 10:1, 11:1, about 12:1, about 13:1, about 14:1, about 15:1, about 16:1, about 17:1, about 18:1, about 19:1, about 20:1, about 21:1, about 22:1, about 23:1, about 24:1, about 25:1, about 26:1, about 27:1, about 28:1, about 29:1, about 30:1, about 50:1, about 100:1, about 150:1, about 200:1, about 250:1, about 300:1).

[0092] In some embodiments, the method further comprises filtering a portion of particles in the suspension through a filter having a pore size of at least 25 pm, more preferably 75 pm. In some embodiments, the filter has a pore size of about 100 pm. In some embodiments, the filter has a pore size of about 75 pm. In some embodiments, the filter has a pore size of about 50 pm. In some embodiments, the filter has a pore size of about 25 pm.

[0093] In some embodiments, the filter has a pore size of about 100 pm or less. In some embodiments, the filter has a pore size of about 75 pm or less. In some embodiments, the filter has a pore size of about 50 pm or less. In some embodiments, the filter has a pore size of about 25 pm or less.

[0094] In some embodiments, the filter has a pore size of about 100 pm or greater. In some embodiments, the filter has a pore size of about 75 pm or greater. In some embodiments, the filter has a pore size of about 50 pm or greater. In some embodiments, the filter has a pore size of about 25 pm or greater.

[0095] In some embodiments, methods of the present disclosure further comprise adding a flocculation agent to the suspension. In certain methods, the flocculation agent is added as an excipient to the aqueous feed solution. In certain other methods, the flocculation agent is added to the carrier liquid, the dehydration liquid, the organic liquid, or any combination thereof.284248642. vl5726.1016001

[0096] In some embodiments, methods of the present disclosure further comprise concentrating the particles, the concentrating comprising performing cross-flow filtration. In some embodiments, methods of the present disclosure further comprise concentrating the particles after suspending the particles in a pharmaceutically acceptable carrier liquid.

[0097] In some embodiments, methods of the present disclosure further comprise concentrating the suspension in a concentration vessel. In certain embodiments, the suspension is mixed and a concentration measurement is taken to determine a concentration of the suspension. An amount of permeate to remove may be calculated to meet a concentration target. The suspension may be circulated through a crossflow filter with a specified pore size at a fixed flow rate. A transmembrane pressure may be controlled and a permeate flow rate may be monitored during filtration. As the suspension is circulated, the carrier liquid may permeate through the filter but not the particles comprising at least one therapeutic agent. As the suspension is filtered by the cross-flow filter, the suspension’s concentration of particles may increase as a measurement of g / cm3.

[0098] In some embodiments, the crossflow filter has a pore size of about 10 pm or less. In some embodiments, the filter has a pore size of about 7.5 pm or less. In some embodiments, the filter has a pore size of about 5.0 pm or less. In some embodiments, the filter has a pore size of about 2.5 pm or less. In some embodiments, the filter has a pore size of about 2.0 pm or less. In some embodiments, the filter has a pore size of about 1.5 pm or less. In some embodiments, the filter has a pore size of about 1.0 pm or less. In some embodiments, the filter has a pore size of about 0.5 pm or less.

[0099] In some embodiments, the crossflow filter has a pore size of about 10 pm. In some embodiments, the filter has a pore size of about 7.5 pm. In some embodiments, the filter has a pore size of about 5.0 pm. In some embodiments, the filter has a pore size of about 2.5 pm. In some embodiments, the filter has a pore size of about 2.0 pm. In some embodiments, the filter has a pore size of about 1.5 pm. In some embodiments, the filter has a pore size of about 1.0 pm. In some embodiments, the filter has a pore size of about 0.5 pm.

[0100] In some embodiments, the crossflow filter has a pore size of about 10 pm or greater. In some embodiments, the filter has a pore size of about 7.5 pm or greater. In some embodiments, the filter has a pore size of about 5.0 pm or greater. In some embodiments, the filter has a pore size of about 2.5 pm or greater. In some embodiments, the filter has a pore size of about 2.0 pm or greater. In some embodiments, the filter has a pore size of about 1.5 pm or greater. In some embodiments, the filter has a pore size of about 1.0 pm or greater. In some embodiments, the filter has a pore size of about 0.5 pm or greater.294248642. vl5726.1016001

[0101] In some embodiments, the crossflow filter may be backpulsed at a fixed pressure at a specified frequency to keep the crossflow filter clear of particles in the suspension maintain permeate flux. The concentration tank may be mixed throughout the crossflow filtering process to maintain homogeneity in the concentration tank.

[0102] In some embodiments, the concentration of particles in the suspension may be increased using centrifugation, a settling tank, a hydrocyclone, or a dead end filter without departing from the scope of the present disclosure.

[0103] In some embodiments, methods of the present disclosure further comprise pumping a suspension from a fill-finish vessel into at least one vial, cartridge, or pre-filled syringe. In certain embodiments, the pumping is peristaltic pumping.

[0104] In some embodiments, methods of the present disclosure further comprise quantifying a concentration of particles in a suspension using nuclear magnetic resonance. In other embodiments, methods of the present disclosure further comprise quantifying a concentration of particles in a fill / finish vessel using nuclear magnetic resonance. In other embodiments, methods of the present disclosure further comprise quantifying a concentration of particles in a suspension pumped from the fill-finish vessel using nuclear magnetic resonance. In some embodiments, the nuclear magnetic resonance quantifies a concentration of particles in a suspension within an at least one vial, cartridge, or pre-filled syringe.

[0105] In some embodiments, quantifying a concentration of a suspension of particles comprises obtaining a relaxation rate of a suspension. In some embodiments, quantifying a concentration of a suspension of particles further comprises obtaining a relaxation rate of a carrier liquid. In some embodiments, quantifying a concentration of a suspension of particles further comprises obtaining a relaxation number from a relaxation rate of the suspension and a relaxation rate of a carrier liquid.

[0106] In some embodiments, an aseptic process comprises collecting particles and combining the particles with a carrier liquid in a closed system without forming a powder intermediate.

[0107] In some embodiments, the particles may be formed and removed from the dehydration liquid to form a mostly dry powder. The mostly dry powder may be further dehydrated by washing the particles with the dehydration liquid once, twice, three times, or more. The mostly dry powder may be dried using a fluidized bed dryer to form a dry powder comprising at least one therapeutic.

[0108] In some embodiments disclosed herein are methods of making a suspension of particles comprising a therapeutic agent, the methods comprising:304248642. vl5726.1016001a) providing a first liquid stream comprising an aqueous feed solution, the feed solution comprising the therapeutic agent;b) providing a second liquid stream comprising an organic liquid;c) dispersing the first liquid stream into the second liquid stream at a first junction;d) applying turbulent flow conditions to a fluid channel connected to the first junction and a second junction, the fluid channel containing the first liquid stream and the second liquid stream, to form aqueous droplets comprising the therapeutic agent;e) dehydrating an aqueous fraction of the aqueous droplets, the dehydrating comprising contacting the aqueous droplets with a dehydration liquid to form particles comprising the therapeutic agent;f) exchanging the dehydration liquid with a carrier liquid aseptically to form the suspension of particles; andg) sparging the suspension of particles to reach a predetermined volume-to- weight percent water content in the particles.

[0109] In some embodiments, the volume-to-weight percent water content in the particles is between about 5% and about 20% (w / v).

[0110] In some embodiments, methods of the present disclosure further comprises filtering out a plurality or a portion of particles from a suspension of particles. In some embodiments, the volume-to-weight percent water content in the particles after filtering out a plurality or a portion of particles from a suspension of particles is between about 0% and about 10% by weight.

[0111] In some embodiments, step g) comprises suspending the particles directly into the carrier liquid without forming a powder intermediate.

[0112] In some embodiments, step a) comprises performing tangential flow filtration to exchange an initial excipient profile of the first liquid stream with a second excipient profile.

[0113] In some embodiments, the first junction includes a rotor-stator mixer, a Tee-mixer, or a static mixer. In some embodiments, the second junction includes a rotor-stator mixer, a Tee-mixer, or a static mixer.

[0114] In some embodiments, the particles may be collected on a membrane.

[0115] In some embodiments, a volume-to-weight percent water content (e.g., predetermined water content (w / v)) in a plurality of particles in the suspension before step g) is between about 5% and 20% (w / v). In some embodiments, an average water content (w / v) in 314248642. vl5726.1016001a plurality of the particles in the suspension after step g) is between about 0% and 10% (w / v). In certain embodiments, an average water content (w / v) in a plurality of the particles in the suspension after step g) is between about 0.1% and 10% (w / v). In still other embodiments, an average water content (w / v) in a plurality of the particles in the suspension after step g) is less than about 50% by weight, less than about 40% (w / v), less than about 30% (w / v), less than about 25% (w / v), less than about 20% (w / v), less than about 15% (w / v), less than about 10% (w / v), less than about 5% (w / v), less than about 3% (w / v), less than about 1% (w / v). In other embodiments, an average water content (w / v) in a plurality of the particles in the suspension after step g) is greater than about 0.1% (w / v).

[0116] In some embodiments, combining the particles with a carrier liquid (e.g., propylene glycol diesters, ethyl oleate, medium chain triglycerides, triacetin, benzyl benzoate, or a combination thereof) aseptically to form the suspension of particles comprises suspending the particles directly into the carrier liquid without forming a powder intermediate.

[0117] In some embodiments, methods of the present disclosure further comprise performing tangential flow filtration to exchange an initial excipient profile of a first liquid stream with a second excipient profile.

[0118] In some embodiments, methods of the present disclosure further comprise increasing the concentration of particles in a suspension. The concentration may be increased by crossflow filtration, a settling tank, centrifugation, hydrocyclones, or any combination thereof.

[0119] In some embodiments, the system may include at least one pump fluidly connected to the fluid channel for adding / removing pressure from the system. In certain embodiments, the first liquid stream and the second liquid stream may each be pressurized by at least one pump to generate the flow rates described herein.Droplets

[0120] Droplets as described herein, can be formed through the input of mechanical energy on a liquid. In some embodiments, droplets of a liquid are formed by a process that imparts high shear stress and / or turbulence (e.g., turbulent flow) on the liquid. The droplets may be formed under conditions of mesomixing (the turbulent exchange between an aqueous liquid stream and its surroundings such as an organic liquid). In certain embodiments, droplets of an aqueous liquid are produced at or after a junction in which a first liquid stream comprising an aqueous feed solution and a second liquid stream comprising an organic liquid are contacted, collide or are combined. In some embodiments, the droplets are formed at the junction, or as324248642. vl5726.1016001the result of turbulent eddies applying shearing forces on the aqueous feed solution within a fluid channel or fluid vessel comprising the aqueous feed solution and the organic liquid, or a combination thereof. In certain embodiments, droplets of an aqueous feed solution are formed as the result of turbulent flow of the first liquid stream and the second liquid stream in a fluid channel or a fluid vessel. In certain other embodiments, droplets of an aqueous feed solution are formed as the result of turbulent flow of the first liquid stream and the second liquid stream in a junction. In some embodiments, droplets of an aqueous feed solution begin forming in a junction and continue forming as the aqueous liquid flows within a fluid channel or a fluid vessel wherein the aqueous liquid is subjected to turbulence.

[0121] As used herein, the term “dispersion,” with respect to droplets of a first liquid, refers to the distribution of the droplets within a second liquid. In some embodiments, a dispersion of droplets of a first liquid within a second liquid is provided, where the typical inter-droplet spacing and droplet number concentration are fairly uniform. Such dispersions may be stable due to the presence of components having both hydrophilic and hydrophobic sites, e.g., as in a surfactant or emulsifier. The terms “dispersed phase” (DP) and “continuous phase” (CP) are related to a dispersion system, in which an aqueous liquid is dispersed within an organic liquid. In such a dispersion system, the term “dispersed phase” (DP) refers aqueous droplets dispersed in the organic liquid. The term “continuous phase” (CP) refers to the organic liquid surrounding the aqueous droplets, e.g., the dispersed phase. As used herein, the term “emulsion” refers to a heterogeneous system consisting of a continuous phase and a non-continuous phase, e.g., the dispersed phase, capable of forming droplets in the continuous phase (CP). The term “emulsifier” refers to an agent that can reduce and / or eliminate the surface and the interfacial tension in a two-phase system. The emulsifier agent may possess both hydrophilic and lipophilic groups. The emulsifier may be considered to be either in the continuous phase (CP), dispersed phase (DP), or both. In some embodiments, the preparation contains excipients. In other embodiments, the preparation further contains a buffer. In certain embodiments, the preparation further contains a surfactant.

[0122] As described herein, in liquid-liquid or solid-liquid dispersions (emulsions or suspensions, respectively), the dispersed phase (DP) is present as discrete droplets or particles which are distributed throughout the continuous phase (CP).

[0123] In some embodiments, the aqueous feed solution or aqueous liquid is water, 0.9% saline, lactated Ringer’s solution, a buffer, dextrose 5%, or a combination thereof. In embodiments, the aqueous feed solution or aqueous liquid is water. In other embodiments, the buffer is acetate buffer, histidine buffer, succinate buffer, HEPES buffer, tris buffer, carbonate 334248642. vl5726.1016001buffer, citrate buffer, phosphate buffer, phosphate-buffered saline, glycine buffer, barbital buffer, cacodylate buffer, ammonium formate buffer, urea solution, or a combination thereof.

[0124] In other embodiments, the aqueous feed solution or aqueous liquid further comprises a carbohydrate, a pH adjusting agent, a salt, a chelator, a mineral, a polymer, a protein stabilizer, an emulsifier, an antiseptic, an amino acid, an antioxidant, a protein, an organic solvent, a paraben, a bactericide, a fungicide, a vitamin, a preservative, a nutrient media, an oligopeptide, a biologic excipient, a chemical excipient, a surfactant, or a combination thereof.

[0125] In certain embodiments, the carbohydrate is dextran, trehalose, sucrose, agarose, mannitol, lactose, sorbitol, maltose, or a combination thereof. In embodiments, the carbohydrate is trehalose, cyclodextrins, hydroxypropyl beta-cyclodextrin, sucrose, sulfobutylether beta-cyclodextrin, or a combination thereof.

[0126] In some embodiments, the pH adjusting agent is acetate, citrate, glutamate, glycinate, histidine, lactate, maleate, phosphate, succinate, tartrate, bicarbonate, aluminum hydroxide, phosphoric acid, hydrochloric acid, DL-lactic / glycolic acids, phosphorylethanolamine, tromethamine, imidazole, glyclyglycine, monosodium glutamate, sodium hydroxide, potassium hydroxide, or a combination thereof. In other embodiments, the pH adjusting agent is citrate, histidine, phosphate, succinate, sodium hydroxide, potassium hydroxide, or a combination thereof. In certain embodiments, the pH adjusting agent is hydrochloric acid or citric acid.

[0127] In other embodiments, the salt is sodium chloride, calcium chloride, potassium chloride, sodium hydroxide, stannous chloride, magnesium sulfate, sodium glucoheptonate, sodium pertechnetate, guanidine hydrochloride, potassium hydroxide, magnesium chloride, potassium nitrate, or a combination thereof. In embodiments, the salt is sodium chloride.

[0128] In certain embodiments, the protein stabilizer is trehalose, polyethylene glycol (PEG), polyoxamers, polyvinylpyrrolidone, polyacrylic acids, poly(vinyl) polymers, polyesters, polyaldehydes, tert-polymers, polyamino acids, hydroxy ethyl starch, N-methyl-2-pyrrolidone, sorbitol, sucrose, mannitol, cyclodextrin, hydroxypropyl beta-cyclodextrin, sulfobutylether beta-cyclodextrin, pentetic acid, or a combination thereof. In embodiments, the protein stabilizer is trehalose, cyclodextrin, hydroxypropyl beta-cyclodextrin, sulfobutylether beta-cyclodextrin, pentetic acid, or a combination thereof. In certain embodiments, the PEG is PEG 200, PEG 300, PEG 3350, PEG 8000, PEG 10000, PEG 20000, or a combination thereof.

[0129] In some embodiments, the emulsifier is polysorbate 80, polysorbate 60, polysorbate 20, sorbitan monooleate, ethanolamine, polyoxyl 35 castor oil, poloxyl 40 hydrogenated castor 344248642. vl5726.1016001oil, carbomer 1342, a corn oil-mono-di -triglyceride, a poly oxy ethylated oleic glyceride, a poloxamer, or a combination thereof. In embodiments, the fatty acid ester of sorbitol is a sorbitan ester, e.g., span 20, span 40, span 60, or span 80. In certain embodiments, the emulsifier is polysorbate 80, sorbitan monooleate, or a combination thereof.

[0130] In other embodiments, the amino acid is alanine, aspartic acid, cysteine, isoleucine, glutamic acid, leucine, phenylalanine, pyrrolysine, serine, selenocysteine, threonine, tryptophan, tyrosine, valine, asparagine, arginine, histidine, glycine, glutamine, proline, methionine, or a combination thereof. In certain embodiments, the amino acid is arginine, histidine, proline, asparagine, or a combination thereof. In embodiments, the amino acid is histidine.

[0131] In some embodiments, the surfactant is polysorbate, magnesium stearate, sodium dodecyl sulfate, TRITON™ N-101, glycerin, poly oxy ethylated castor oil, docusate, sodium stearate, decyl glucoside, nonoxynol-9, cetyltrimethylammonium bromide, sodium bis(2-ethylhexyl) sulfosuccinate, lecithin, sorbitan ester, phosphatidylcholine, polyglycerol polyricinoleate, siloxanes, cetyl polyethylene glycol / polypropylene glycol- 10 / 1 dimethicone triglyceride, bis- polyethylene glycol / polypropylene glycol-14 / 14 dimethicone, bis-(glyceryl / lauryl) glyceryl lauryl dimethicone and caprylic / capric triglyceride, cetyl polyethylene glycol / polypropylene glycol- 10 / 1 dimethicone, phospholipids, or a combination thereof. In other embodiments, the surfactant is polysorbate, docusate or lecithin. In certain embodiments, the surfactant is polysorbate 20, polysorbate 60, or polysorbate 80, e.g., Tween 20, Tween 60, Tween 80. In still other embodiments, the surfactant is polysorbate 20 or polysorbate 80. In certain embodiments, the fatty acid ester of sorbitol is a sorbitan ester, e.g., span 20, span 40, span 60, or span 80. In other embodiments, the surfactant is an ionic surfactant. In embodiments, the surfactant is polysorbate 80.

[0132] In other embodiments, the concentration of an therapeutic agent in the aqueous feed solution as described herein, is about 10 mg / mL to about 650 mg / mL, e.g., about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 525, 550, 575, 600, 625 mg / mL to about 650 mg / mL; about 20 mg / mL to about 625 mg / mL, e.g., about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 525, 550, 575, 600 mg / mL to about 625 mg / mL; about 20 mg / mL to about 600 mg / mL, e.g., about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 525, 550, 575 mg / mL to about 600 mg / mL; about 20 mg / mL to about 575 mg / mL, e.g., about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 525, 550 mg / mL to about 575 mg / mL; about 20 mg / mL to about 354248642. vl5726.1016001550 mg / mL, e.g, about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 525 mg / mL to about 550 mg / mL; about 20 mg / mL to about 525 mg / mL, e.g., about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500 mg / mL to about 525 mg / mL; about 20 mg / mL to about 500 mg / mL, e.g., about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475 mg / mL to about 500 mg / mL; about 20 mg / mL to about 475 mg / mL, e.g., about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450 mg / mL to about 475 mg / mL; about 20 mg / mL to about 450 mg / mL, e.g., about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425 mg / mL to about 450 mg / mL; about 20 mg / mL to about 425 mg / mL, e.g., about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400 mg / mL to about 425 mg / mL; about 20 mg / mL to about 400 mg / mL, e.g., about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375 mg / mL to about 400 mg / mL; about 20 mg / mL to about 375 mg / mL, e.g., about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350 mg / mL to about 375 mg / mL; about 20 mg / mL to about 350 mg / mL, e.g., about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325 mg / mL to about 350 mg / mL; about 20 mg / mL to about 325 mg / mL, e.g., about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300 mg / mL to about 325 mg / mL; or about 20 mg / mL to about 300 mg / mL, e.g., about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275 mg / mL to about 300 mg / mL. In some embodiments, a therapeutic agent has a concentration of from about 70 g / L to about 160 g / L in an aqueous feed solution.

[0133] In certain embodiments of the present disclosure, the concentration of a therapeutic agent in an aqueous feed solution is from about 0.0001 mg / mL to about 1000 mg / mL, e.g., about 100 to about 800, about 200 to about 700, about 200 to about 600, or about 300 mg / mL to about 700 mg / mL. In other embodiments, the concentration of a therapeutic agent in an aqueous feed solution is about 10 mg / mL to about 500 mg / mL. In still other embodiments, the concentration of a therapeutic agent in an aqueous feed solution is about 10 mg / mL to about 100 mg / mL. In embodiments, the concentration of a therapeutic agent in an aqueous feed solution is about 20 mg / mL to about 100 mg / mL. In some embodiments, the particles have a mass loading of a therapeutic agent from about 1% to about 100%.

[0134] In other embodiments, the concentration of a therapeutic agent in a first liquid stream is about 10 mg / mL to about 650 mg / mL, e.g., about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 525, 550, 575, 600, 625 mg / mL to about 650 mg / mL; about 20 mg / mL to about 625 mg / mL, e.g., about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 525, 550, 575,364248642. vl5726.1016001600 mg / mL to about 625 mg / mL; about 20 mg / mL to about 600 mg / mL, e.g., about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 525, 550, 575 mg / mL to about 600 mg / mL; about 20 mg / mL to about 575 mg / mL, e.g., about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 525, 550 mg / mL to about 575 mg / mL; about 20 mg / mL to about 550 mg / mL, e.g., about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 525 mg / mL to about 550 mg / mL; about 20 mg / mL to about 525 mg / mL, e.g., about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500 mg / mL to about 525 mg / mL; about 20 mg / mL to about 500 mg / mL, e.g., about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475 mg / mL to about 500 mg / mL; about 20 mg / mL to about 475 mg / mL, e.g., about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450 mg / mL to about 475 mg / mL; about 20 mg / mL to about 450 mg / mL, e.g., about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425 mg / mL to about 450 mg / mL; about 20 mg / mL to about 425 mg / mL, e.g., about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400 mg / mL to about 425 mg / mL; about 20 mg / mL to about 400 mg / mL, e.g., about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375 mg / mL to about 400 mg / mL; about 20 mg / mL to about 375 mg / mL, e.g., about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350 mg / mL to about 375 mg / mL; about 20 mg / mL to about 350 mg / mL, e.g., about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325 mg / mL to about 350 mg / mL; about 20 mg / mL to about 325 mg / mL, e.g., about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300 mg / mL to about 325 mg / mL; or about 20 mg / mL to about 300 mg / mL, e.g., about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275 mg / mL to about 300 mg / mL. In some embodiments, a therapeutic agent has a concentration of from about 70 g / L to about 160 g / L in a first liquid stream.

[0135] In other embodiments, the aqueous liquid has a viscosity of less than about 200 mPa s, less than about 150 mPa s, less than about 125 mPa s, less than about 100 mPa s, less than about 75 mPa s, less than about 75 mPa s, less than about 70 mPa s, less than about 65 mPa s, less than about 60 mPa s, less than about 55 mPa s, less than about 50 mPa s, less than about 45 mPa s, less than about 40 mPa s, less than about 35 mPa s, less than about 30 mPa s, less than about 25 mPa s, less than about 20 mPa s, less than about 19 mPa s, less than about 18 mPa s, less than about 17 mPa s, less than about 16 mPa s, less than about 15 mPa s, less than about 14 mPa s, less than about 13 mPa s, less than about 12 mPa s, less than about 11 mPa s, less than about 10 mPa s, less than about 9.5 mPa s, less than about 9 mPa s, less than 374248642. vl5726.1016001about 8.5 mPa s, less than about 8 mPa s, less than about 7.5 mPa s, less than about 7 mPa s, less than about 6.5 mPa s, less than about 6 mPa s, less than about 5.5 mPa s, less than about 5 mPa s, less than about 4.5 mPa s, less than about 4 mPa s, less than about 3.5 mPa s, less than about 3 mPa s, less than about 2.5 mPa s, less than about 2 mPa s, less than about 1.5 mPa s, less than about 1 mPa s, or less than about 0.5 mPa s (one millipascal-second).

[0136] The term “viscosity” is used to describe the property of a fluid acting to resist shearing flow. For the purposes of the present disclosure, viscosity can be determined using a rheometer, e.g., AR-G2 Rheometer (TA Instruments, USA), fitted with a cone and plate (2° / 40 mm) at 25 °C at a specified shear rate. In certain embodiments, the viscosity is measured at a shear rate in the Newtonian regime. The term “Newtonian regime” means a range of shear rates which are linearly proportional or nearly linearly proportional to the local strain rate at every point. In some embodiments, the viscosity is measured at a shear rate of about 100 s'1or greater, e.g., at about 1000 s'1or greater than about 1000 s'1. Methods of controlling viscosity include temperature regulation and viscosity modifying additives. Mixtures of liquids may also be used to control viscosity. The units “mPa s” and “cP” are used herein, interchangeably in the broadest sense.

[0137] In some embodiments, the aqueous liquid has a viscosity of less than about 100 mPa s. In other embodiments, the aqueous liquid has a viscosity of less than about 10 mPa s. In certain other embodiments, the aqueous liquid has a viscosity of less than about 3 mPa s. In still other embodiments, the aqueous liquid has a viscosity of less than about 0.9 mPa s. In embodiments, the aqueous liquid has a viscosity of less than about 0.5 mPa s.

[0138] In embodiments, the organic liquid is an organic solvent. In some embodiments, the organic liquid is an ester or an alcohol. In some embodiments, the dehydration liquid is an organic solvent. In some embodiments, the dehydration liquid is an ester or an alcohol.

[0139] In some embodiments, the organic solvent is acetone, acetonitrile, acyclic alkanes (e.g., hexanes, heptane, pentane), amyl acetate, butanol, butyl acetate, chlorobenzene, chloroform, cumene, cyclohexane, 1,2-di chloroethene, di chloromethane, diethyl ether, dimethoxy ethane, dimethylacetamide, dimethylformamide, dimethyl sulfoxide, 1,4-di oxane, ethanol, 2-ethoxyethanol, ethyl acetate, ethyl nitrate, ethyleneglycol, hydrazine, isopropanol, methanol, methyl acetate, 2 -m ethyl- 1 -butanol, 2-methyl-l -propanol, methylbutyl ketone, methylcyclohexane, methylethyl ketone, methylpyrrolidone, methyl tert-butyl ether, nitromethane, propanol, propyl acetate, sulfolane, propyleneglycol, tetrahydrofuran, tetralin, toluene, 1,1,2-tricholoroethane, tri ethylamine, xylene, benzyl benzoate, ethyl lactate, dimethyl isosorbide, dimethyl sulfoxide, glycofurol, diglyme, methyl tert-butyl ether, polyethylene 384248642. vl5726.1016001glycol, 2-pyrrolidone, tetrahydrofurfuryl alcohol, trigylcerides, octyl acetate, ethanol, butanol, octanol, decanol, diglyme, tocopherol, octa-fluoropropane, (perfluorohexyl)octane, n-acetyltryptophan, trigylcerides, triglycerides of the fractionated plant fatty acids C8 and CIO, propylene glycol diesters of saturated plant fatty acids C8 and CIO, propylene glycol diesters of Di caprate / Di capryl ate (C8 and CIO), propylene glycol diesters of Dicaprylate (C8), propylene glycol diesters of Dicaprate (CIO), ethyl laurate, methyl caprylate, ethyl laurate, ethyl myristate, ethyl caprylate, methyl caprate, methyl myristate, methyl oleate, methyl linoleate, dimethyl adipate, dibutyl suberate, diethyl sebacate, ethyl macadamiate, trimethylolpropane triisosterate, isopropyl laurate, isopropyl myristate, diethyl succinate, polysorbate esters, ethanol amine, propanoic acid, triacetin, citral, anisole, anethol, benzaldehyde, linalool, caprolactone, phenol, thioglycerol, dimethylacetamide, ethyl formate, ethyl hexyl acetate, eugenol, clove bud oil, diethyl glycol monoether, dimethyl isosorbide, isopropyl acetate, methyl isobutyl ketone, methyl tert-butyl ether, N-methyl pyrrolidone, perfluorodecalin, 2-pyrrolidone, ethyl oleate, ethyl caprate, dibutyl adipate, hexanoic acid, octanoic acid, diethyl glycol monoether, gamma-butyrolactone, polyoxyl 40 hydrogenated castor oil, polyoxyl 35 castor oil, propylene carbonate, octanol, hexanol, sorbitan monooleate, n-acetyltryptophan, solketal, an alkyl acetate, an aryl acetate, an aryl alkyl acetate, tolyl acetate, benzyl acetate, polysorbate 80, phenethyl acetate, phenyl acetate, glycerol, or a combination thereof. In certain embodiments, the organic solvent is a Class III solvent as defined by the International Council for Harmonisation (ICH) guidelines. In some embodiments, the organic liquid is n-butyl acetate or pentanol. In some embodiments, the organic liquid is n-butyl acetate. In some embodiments, the organic liquid is hydrophobic. In certain embodiments, the organic liquid is immiscible with an aqueous liquid.

[0140] In some embodiments, the organic solvent is benzyl alcohol, benzyl benzoate, castor oil, coconut oil, com oil, cottonseed oil, fish oil, grape seed oil, hazelnut oil, hydrogenated palm seed 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, methyl acetate, ethyl acetate, propyl acetate, butyl acetate, ethyl ether, ethyl lactate, isopropyl acetate, methyl acetate, methyl isobutyl ketone, methyl tert-butyl ether, N-methyl pyrrolidone, perfluorodecalin, 2-pyrrolidone, trigylcerides, tetrahydrofurfuryl alcohol, triglycerides of the fractionated plant fatty acids C8 and CIO (e.g., MIGLYOL® 810 and MIGLOYL® 812N), propylene glycol diesters of saturated plant fatty acids C8 and CIO (e.g., MIGLYOL® 840), propylene glycol diesters of Dicaprate / Dicaprylate (C8 and CIO), propylene glycol diesters of 394248642. vl5726.1016001Dicaprylate (C8), propylene glycol diesters of Dicaprate (CIO), 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, hexanes, heptane, or a combination thereof. In some embodiments, the organic solvent is hydrophobic. In certain embodiments, the organic solvent is immiscible with an aqueous liquid. In some embodiments, the dehydration liquid is an organic solvent. In certain embodiments, the dehydration liquid is immiscible with the aqueous liquid.

[0141] In certain embodiments, the organic solvent is acetonitrile, chlorobenzene, chloroform, cyclohexane, cumene, 1,2-di chloroethene, di chloromethane, 1,2-dimethoxy ethane, N,N-dimethylacetamide, N,N-dimethylformamide, 1,4-di oxane, 2-ethoxyethanol, ethyleneglycol, formamide, hexane, methanol, 2-methoxyethanol, methylbutyl ketone, methylcyclohexane, methylisobutylketone, N-methylpyrrolidone, nitromethane, pyridine, sulfolane, tetrahydrofuran, tetralin, toluene, 1,1,2-trichloroethene, xylene, acetic acid, acetone, anisole, 1 -butanol, 2-butanol, butylacetate, tert-butylmethyl ether, dimethyl sulfoxide, ethanol, ethylacetate, ethyl ether, ethyl formate, formic acid, heptane, isobutylacetate, isopropyl acetate, methylacetate, 3 -methyl- 1 -butanol, methylethyl ketone, 2-methyl-l-propanol, pentane, 1 -pentanol, 1 -propanol, 2-propanol, propyl acetate, tri ethylamine, 1,1-di ethoxypropane, 1,1 -dimethoxymethane, 2,2-dimethoxypropane, isooctane, isopropyl ether, methylisopropyl ketone, methyltetrahydrofuran, petroleum ether, trichloroacetic acid, trifluoroacetic acid, decanol, 2-ethylhexylacetate, amylacetate, or a combination thereof. In embodiments, the organic solvent is methylacetate, ethylacetate, propylacetate, butylacetate, amylacetate, 2-ethylhexylacetate, heptane, or a combination thereof.

[0142] In some embodiments, the organic liquid and the dehydration liquid are the same liquid. In some embodiments, the organic liquid and the dehydration liquid are different liquids.

[0143] In some embodiments, the organic liquid further comprises a carbohydrate, a pH adjusting agent, a salt, a chelator, a mineral, a polymer, a protein stabilizer, an emulsifier, an antiseptic, an amino acid, an antioxidant, a protein, an organic solvent, a paraben, a bactericide, a fungicide, a vitamin, a preservative, a nutrient media, an oligopeptide, a biologic excipient, a chemical excipient, a surfactant, a flocculation agent, or a combination thereof. In embodiments, the organic liquid further comprises a surfactant.

[0144] In other embodiments, the surfactant is polysorbate, magnesium stearate, sodium dodecyl sulfate, TRITON™ N-101, glycerin, poly oxy ethylated castor oil, docusate, sodium stearate, decyl glucoside, nonoxynol-9, cetyltrimethylammonium bromide, sodium bis(2-ethylhexyl) sulfosuccinate, lecithin, sorbitan ester, phosphatidylcholine, polyglycerol 404248642. vl5726.1016001polyricinoleate, siloxanes, cetyl polyethylene glycol / polypropylene glycol- 10 / 1 dimethicone triglyceride, bis- polyethylene glycol / polypropylene glycol-14 / 14 dimethicone, bis-(glyceryl / lauryl) glyceryl lauryl dimethicone and caprylic / capric triglyceride, cetyl polyethylene glycol / polypropylene glycol- 10 / 1 dimethicone, phospholipids, or a combination thereof. In certain embodiments, the surfactant is polysorbate, docusate or lecithin. In certain other embodiments, the surfactant is polysorbate 20, polysorbate 60, or polysorbate 80, e.g, Tween 20, Tween 60, Tween 80. In still other embodiments, the surfactant is polysorbate 20 or polysorbate 80. In certain embodiments, the fatty acid ester of sorbitol is a sorbitan ester, e.g, span 20, span 40, span 60, or span 80. In other embodiments, the surfactant is an ionic surfactant. In embodiments, the surfactant is polysorbate 80.

[0145] In some embodiments, the flocculation agent is ionic or non-ionic. Generally, an ionic flocculation agent interacts with charged particles, e.g., anionic flocculation agent interacts with positively charged particles, and a cationic flocculation agent interacts with negatively charged particles. In some embodiments, the flocculation agent is ionic. In some embodiments, the ionic flocculation agent is magnesium stearate, sodium dodecyl sulfate, sodium stearate, cetyltrimethyl ammonium bromide, lecithin, or a combination thereof. In particular embodiments, the flocculation agent is non-ionic. As disclosed herein, a non-ionic flocculation agent interacts with charged particles. In certain embodiments, the non-ionic flocculation agent is a polysorbate (polysorbate 80, polysorbate 60, polysorbate 20, e.g., Tween 80, Tween 60, Tween 20), an alkylphenol ethoxylate, glycerol, polyoxyethylated castor oil, docusate, decyl glucoside, nonoxynol-9, a sorbitan ester, sorbitan monooleate, ethanolamine, polyoxyl 35 castor oil, poloxyl 40 hydrogenated castor oil, carbomer 1342, a com oil-mono-di-triglyceride, a polyoxyethylated oleic glyceride, a poloxamer, or a combination thereof. In some embodiments, the non-ionic flocculation agent is a polysorbate, an alkylphenol ethoxylate, a sorbitan ester, a poloxamer, or a combination thereof. In certain embodiments, the non-ionic flocculation agent is a polysorbate or a sorbitan ester.

[0146] In some embodiments, the organic liquid has a viscosity of less than about 200 mPa s, less than about 150 mPa s, less than about 125 mPa s, less than about 100 mPa s, less than about 75 mPa s, less than about 75 mPa s, less than about 70 mPa s, less than about 65 mPa s, less than about 60 mPa s, less than about 55 mPa s, less than about 50 mPa s, less than about 45 mPa s, less than about 40 mPa s, less than about 35 mPa s, less than about 30 mPa s, less than about 25 mPa s, less than about 20 mPa s, less than about 19 mPa s, less than about 18 mPa s, less than about 17 mPa s, less than about 16 mPa s, less than about 15 mPa s, less than about 14 mPa s, less than about 13 mPa s, less than about 12 mPa s, less than about 11414248642. vl5726.1016001mPa s, less than about 10 mPa s, less than about 9.5 mPa s, less than about 9 mPa s, less than about 8.5 mPa s, less than about 8 mPa s, less than about 7.5 mPa s, less than about 7 mPa s, less than about 6.5 mPa s, less than about 6 mPa s, less than about 5.5 mPa s, less than about 5 mPa s, less than about 4.5 mPa s, less than about 4 mPa s, less than about 3.5 mPa s, less than about 3 mPa s, less than about 2.5 mPa s, less than about 2 mPa s, less than about 1.5 mPa s, less than about 1 mPa s, less than about 0.5 mPa s, less than about 0.1 mPa s, less than about 0.05 mPa s, or less than about 0.01 mPa s (one millipascal-second). Methods of controlling viscosity include temperature regulation and viscosity modifying additives. Mixtures of liquids may also be used to control viscosity.

[0147] In other embodiments, the organic liquid has a viscosity of less than about 50 mPa- s. In some embodiments, the organic liquid has a viscosity of less than about 10 mPa s. In certain other embodiments, the organic liquid has a viscosity of less than about 5 mPa s. In still other embodiments, the organic liquid has a viscosity of less than about 2 mPa s.

[0148] In other embodiments as described herein, methods of the present disclosure further comprises decreasing the temperature of the organic liquid to a temperature within about 30 °C of the freezing point of the aqueous feed solution. In some embodiments, the boiling point of the organic liquid at atmospheric pressure is from about 0 to about 200 °C. In certain embodiments the temperature, pressure, and water content of the organic liquid, in which the aqueous droplets are dispersed can be regulated to control the dehydration kinetics.

[0149] In some embodiments static mixing includes one or more of: membrane emulsification, homogenization, mechanical stirring, mechanical shaking, impinging jet mixing, ultra-sound, sonication, micro-channel emulsification, microsieve emulsification, or capillary extrusion. In certain embodiments, the micro-channel emulsification is accomplished using a microfluidic chip based device. In certain other embodiments, the membrane emulsification is conducted by rotating membrane emulsification, cross-flow membrane emulsification, or a combination thereof. In still other embodiments, the homogenization is conducted by shear homogenization, pressure homogenization, rotor-stator homogenization, microfluidization, or a combination thereof. In some embodiments, static mixing includes forming a turbulent flow in a fluid channel or a fluid vessel of an aqueous liquid and an organic liquid to introduce or form turbulent eddies within the fluid channel or fluid vessel. The turbulent eddies may apply a shearing force on the aqueous feed solution within the fluid channel to form aqueous droplets of the aqueous feed solution within the organic liquid. A person of ordinary skill in the field of this disclosure can readily assess the shear homogenization or pressure homogenization of the disclosed methods using routine and 424248642. vl5726.1016001standard techniques for high or how shear homogenization, or high or low pressure homogenization. In certain embodiments, the mechanical stirring is conducted by a turbulent stirred vessel, a magnetic stirring device, a mechanical stirring device, or a combination thereof. In certain embodiments, the static mixing comprises laminar flow, turbulent flow, transition flow, or a combination thereof.

[0150] In some embodiments, dispersing a first liquid stream into a second liquid stream at or after a junction includes flowing the first liquid stream and / or the second liquid stream into a rotor-stator mixer, Tee-mixer, or static mixer at or after the junction. In certain embodiments, the rotor-stator mixer may be used without a junction. A fluid channel or a fluid vessel within the junction or after the junction may include a turbulent flow applying shear forces on an aqueous liquid within the fluid channel. The fluid channel may connect one or more junctions. In some embodiments, a first junction may include a first inlet channel for the first liquid stream and a second inlet channel for the second liquid stream. The first inlet channel and the second inlet channel may join to form the fluid channel. As the first liquid stream flows through the first inlet channel into the first junction and the second liquid stream flows through the second inlet channel into the first junction, the first liquid stream may be dispersed into the second liquid stream. In some embodiments, the first liquid stream may be dispersed into the second liquid using a rotor stator mixer, Tee-mixer, or junction with a static mixer. In certain embodiments, the first liquid stream may be dispersed into the second liquid stream using one or more Tee-mixers or one or more junctions with static mixers. The second liquid stream may include a turbulent flow. The first junction may be fluidly connected to a fluid channel or a fluid vessel. The first liquid stream and the second liquid stream may be immiscible and do not mix in the fluid channel or fluid vessel.

[0151] In some embodiments, the first liquid stream is dispersed into the second liquid stream at a temperature of from about 3°C to about 40°C.

[0152] A turbulent flow may be applied to the fluid channel or the fluid vessel by a rotorstator mixer, a Tee-mixer, or a static mixer at or after the junction. The rotor-stator mixer, Tee-mixer, static mixer, or turbulent flow may apply a shearing force to the first liquid within the fluid channel or the fluid vessel to form aqueous droplets within the second liquid.

[0153] A fluid channel may fluidly connect the first junction to a second junction. In some embodiments, the first junction and the second junction may be connected together directly. The second junction may include a solvent inlet channel for introducing a dehydration liquid into the fluid channel or fluid vessel. The dehydration liquid may dehydrate the aqueous droplets. The dehydration process may occur in a primary dehydration step. A third junction 434248642. vl5726.1016001may connect the fluid channel to a dehydration tube, dehydration vessel, or combination thereof.

[0154] The fluid channel may expand or contract with respect to its internal diameter at each junction.

[0155] In some embodiments, the static mixing comprises using a Tee-mixer or a Y-mixer. In certain embodiments, the Tee-mixer is a side Tee mixer. In certain other embodiments, the Tee-mixer is a multi-jet design.

[0156] In some embodiments, the static mixer comprises a motionless mixer; plate-type mixer; vortex design mixer; helical static mixer; HSM high shear mixer; wafer type static mixer; fin type static mixer; KMS, KME, KMX, HEV, SMV, SMX, SMXL, SMR, KVM, KHT, SMF, or KFBE mixer offered by Chemineer, Inc. located in Dayton, Ohio; Komax mixer offered by Komax Systems, Inc. located in Huntington Beach, California; ISG mixer offered by Charles Ross & Son Company located in Hauppauge, NY; SMV, SMX, SMXL, SMF, or SMR mixer offered by Sulzer Chemtech located in Winterthur, Switzerland; or any combination thereof. Other static mixers known in the art may be substituted without departing from the scope of the present disclosure.

[0157] In some embodiments, a first liquid stream flow rate is from about 0.1 mL / min to about 500 mL / min (e.g., about 0.1 mL / min to about 500 mL / min, about 0.1 mL / min to about 450 mL / min, about 0.1 mL / min to about 400 mL / min, about 0.1 mL / min to about 350 mL / min, about 0.1 mL / min to about 300 mL / min, about 0.1 mL / min to about 250 mL / min, about 0.1 mL / min to about 200 mL / min, about 0.1 mL / min to about 150 mL / min, about 0.1 mL / min to about 100 mL / min, about 0.1 mL / min to about 50 mL / min, about 0.5 mL / min to about 50 mL / min, about 1 mL / min to about 50 mL / min, about 2 mL / min to about 50 mL / min, about 3 mL / min to about 50 mL / min, about 4 mL / min to about 50 mL / min, about 5 mL / min to about 50 mL / min, about 6 mL / min to about 50 mL / min, about 7 mL / min to about 50 mL / min, about 8 mL / min to about 50 mL / min, about 9 mL / min to about 50 mL / min, about 10 mL / min to about 50 mL / min, about 15 mL / min to about 50 mL / min, about 15 mL / min to about 45 mL / min, about 15 mL / min to about 40 mL / min, about 15 mL / min to about 35 mL / min, about 15 mL / min to about 30 mL / min, about 20 mL / min to about 30 mL / min, etc.). In some embodiments, a first liquid stream flow rate is from about 10 mL / min to about 50 mL / min. In some embodiments, a first liquid stream flow rate is from about 20 mL / min to about 30 mL / min. In some embodiments, a first liquid stream flow rate is greater than about 5 mL / min, greater than about 10 mL / min, greater than about 15 mL / min, greater than about 20 mL / min, greater than about444248642. vl5726.101600125 mL / min, greater than about 30 mL / min, greater than about 35 mL / min, greater than about 40 mL / min, greater than about 45 mL / min, greater than about 50 mL / min.

[0158] In some embodiments, dispersing a first liquid stream into a second liquid stream at or after a junction is performed using a first liquid stream flow rate of from about 0.1 mL / min to about 500 mL / min (e.g., about 0.1 mL / min to about 500 mL / min, about 0.1 mL / min to about 450 mL / min, about 0.1 mL / min to about 400 mL / min, about 0.1 mL / min to about 350 mL / min, about 0.1 mL / min to about 300 mL / min, about 0.1 mL / min to about 250 mL / min, about 0.1 mL / min to about 200 mL / min, about 0.1 mL / min to about 150 mL / min, about 0.1 mL / min to about 100 mL / min, about 0.1 mL / min to about 50 mL / min, about 0.5 mL / min to about 50 mL / min, about 1 mL / min to about 50 mL / min, about 2 mL / min to about 50 mL / min, about 3 mL / min to about 50 mL / min, about 4 mL / min to about 50 mL / min, about 5 mL / min to about 50 mL / min, about 6 mL / min to about 50 mL / min, about 7 mL / min to about 50 mL / min, about 8 mL / min to about 50 mL / min, about 9 mL / min to about 50 mL / min, about 10 mL / min to about 50 mL / min, about 15 mL / min to about 50 mL / min, about 15 mL / min to about 45 mL / min, about 15 mL / min to about 40 mL / min, about 15 mL / min to about 35 mL / min, about 15 mL / min to about 30 mL / min, about 20 mL / min to about 30 mL / min, etc.). In some embodiments, dispersing a first liquid stream into a second liquid stream at or after a junction is performed using a first liquid stream flow rate of from about 10 mL / min to about 50 mL / min. In some embodiments, the first liquid stream flow rate is from about 20 mL / min to about 30 mL / min.

[0159] In some embodiments, dispersing a first liquid stream into a second liquid stream at or after a junction is performed using a second liquid stream flow rate (e.g., turbulent flow rate) of from about 0.1 L / min to about 100 L / min (e.g., about 0.1 mL / min to about 90 mL / min, about 0.1 mL / min to about 80 mL / min, about 0.1 mL / min to about 70 mL / min, about 0.1 mL / min to about 60 mL / min, about 0.1 mL / min to about 50 mL / min, about 0.1 mL / min to about 40 mL / min, about 0.1 mL / min to about 30 mL / min, about 0.1 mL / min to about 20 mL / min, about 0.1 mL / min to about 10 mL / min, about 0.5 mL / min to about 10 mL / min, about 1 mL / min to about 10 mL / min, etc.). In some embodiments, dispersing a first liquid stream into a second liquid stream at or after a junction is performed using a second liquid stream flow rate of from about 0.1 L / min to about 50 L / min. In some embodiments, the second liquid stream flow rate of from about 1 L / min to about 20 L / min. In some embodiments, the second liquid stream flow rate of from about 1 L / min to about 10 L / min.

[0160] In some embodiments, the dispersing a first liquid stream into a second liquid stream at or after a junction is performed using an organic liquid flow rate of from about 0.1 L / min to about 100 L / min (e.g., about 0.1 mL / min to about 90 mL / min, about 0.1 mL / min to 454248642. vl5726.1016001about 80 mL / min, about 0.1 mL / min to about 70 mL / min, about 0.1 mL / min to about 60 mL / min, about 0.1 mL / min to about 50 mL / min, about 0.1 mL / min to about 40 mL / min, about 0.1 mL / min to about 30 mL / min, about 0.1 mL / min to about 20 mL / min, about 0.1 mL / min to about 10 mL / min, about 0.5 mL / min to about 10 mL / min, about 1 mL / min to about 10 mL / min, etc.). In some embodiments, dispersing a first liquid stream into a second liquid stream at or after a junction is performed using an organic liquid flow rate of from about 0.1 L / min to about 50 L / min. In some embodiments, the organic liquid flow rate of from about 1 L / min to about 10 L / min. In some embodiments, dispersing a first liquid stream into a second liquid stream at or after a junction is performed using an organic liquid flow rate greater than about 1 L / min, greater than about 2 L / min, greater than about 3 L / min, greater than about 4 L / min, greater than about 5 L / min, greater than about 6 L / min, greater than about 7 L / min, greater than about 8 L / min, greater than about 9 L / min, greater than about 10 L / min.

[0161] In some embodiments, dispersing a first liquid stream into a second liquid stream at or after a junction (e.g., a first junction, a second junction) is performed using a feed solution to organic liquid flow rate ratio of from about 1:1 to about 1000:1 (e.g., about 5:1 to about 1000:1, about 10:1 to about 1000:1, about 20:1 to about 1000:1, about 50:1 to about 1000:1, about 60: 1 to about 1000: 1, about 70: 1 to about 1000: 1, about 80: 1 to about 1000: 1, about 90: 1 to about 1000: 1, about 100: 1 to about 1000: 1, about 100: 1 to about 900: 1, about 100: 1 to about 800:1, about 100:1 to about 700:1, about 100:1 to about 600:1, about 100:1 to about 500:1, about 25:1 to about 500:1, about 25:1 to about 400:1, about 25:1 to about 300:1, about 25:1 to about 275:1, about 25:1 to about 250:1, about 25:1 to about 225:1, about 25:1 to about 200:1, about 25:1 to about 175:1, about 25:1 to about 150:1, about 25:1 to about 125:1, about 25:1 to about 100:1, about 25:1 to about 75:1, about 25:1 to about 50:1, about 10:1 to about 500:1, about 10:1 to about 400:1, about 10:1 to about 300:1, about 10:1 to about 275:1, about 10:1 to about 250:1, about 10:1 to about 225:1, about 10:1 to about 200:1, about 10:1 to about 175:1, about 10:1 to about 150:1, about 10:1 to about 125:1, about 10:1 to about 100:1, about 10:1 to about 75:1, about 10:1 to about 50:1, about 10:1 to about 10:25, etc.). In some embodiments, dispersing a first liquid stream into a second liquid stream at or after a junction is performed using a feed solution to organic liquid flow rate ratio of from about 1 : 1 to about 500: 1. In some embodiments, the feed solution to organic liquid flow rate ratio is from about 100:1 to about 500:1.

[0162] In some embodiments, dispersing a first liquid stream into a second liquid stream at or after a junction is performed using a first liquid stream flow rate to second liquid stream flow rate ratio of from about 1:1 to about 1000:1 (e.g., about 5:1 to about 1000:1, about 10:1464248642. vl5726.1016001to about 1000:1, about 20:1 to about 1000:1, about 50:1 to about 1000:1, about 60:1 to about 1000:1, about 70:1 to about 1000:1, about 80:1 to about 1000:1, about 90:1 to about 1000:1, about 100:1 to about 1000:1, about 100:1 to about 900:1, about 100:1 to about 800:1, about 100:1 to about 700:1, about 100:1 to about 600:1, about 100:1 to about 500:1, about 25:1 to about 500:1, about 25:1 to about 400:1, about 25:1 to about 300:1, about 25:1 to about 275:1, about 25:1 to about 250:1, about 25:1 to about 225:1, about 25:1 to about 200:1, about 25:1 to about 175:1, about 25:1 to about 150:1, about 25:1 to about 125:1, about 25:1 to about 100:1, about 25:1 to about 75:1, about 25:1 to about 50:1, about 10:1 to about 500:1, about 10:1 to about 400:1, about 10:1 to about 300:1, about 10:1 to about 275:1, about 10:1 to about 250:1, about 10:1 to about 225:1, about 10:1 to about 200:1, about 10:1 to about 175:1, about 10:1 to about 150:1, about 10:1 to about 125:1, about 10:1 to about 100:1, about 10:1 to about 75:1, about 10:1 to about 50:1, about 10:1 to about 10:25, etc.). In some embodiments, dispersing a first liquid stream into a second liquid stream at or after a junction is performed using a first liquid stream flow rate to second liquid stream flow rate ratio of from about 1 : 1 to about 500: 1. In some embodiments, the first liquid stream flow rate to second liquid stream flow rate ratio is from about 100:1 to about 500:1.

[0163] In some embodiments, turbulent flow conditions are applied using a first liquid stream flow rate to second liquid stream flow rate ratio of from about 1 : 1 to about 1000: 1 (e.g., about 5:1 to about 1000:1, about 10:1 to about 1000:1, about 20:1 to about 1000:1, about 50:1 to about 1000:1, about 60:1 to about 1000:1, about 70:1 to about 1000:1, about 80:1 to about 1000:1, about 90:1 to about 1000:1, about 100:1 to about 1000:1, about 100:1 to about 900:1, about 100:1 to about 800:1, about 100:1 to about 700:1, about 100:1 to about 600:1, about 100:1 to about 500:1, about 25:1 to about 500:1, about 25:1 to about 400:1, about 25:1 to about 300:1, about 25:1 to about 275:1, about 25:1 to about 250:1, about 25:1 to about 225:1, about 25:1 to about 200:1, about 25:1 to about 175:1, about 25:1 to about 150:1, about 25:1 to about 125:1, about 25:1 to about 100:1, about 25:1 to about 75:1, about 25:1 to about 50:1, about 10:1 to about 500:1, about 10:1 to about 400:1, about 10:1 to about 300:1, about 10:1 to about 275:1, about 10:1 to about 250:1, about 10:1 to about 225:1, about 10:1 to about 200:1, about 10:1 to about 175:1, about 10:1 to about 150:1, about 10:1 to about 125:1, about 10:1 to about 100:1, about 10:1 to about 75:1, about 10:1 to about 50:1, about 10:1 to about 10:25, etc.). In some embodiments, turbulent flow conditions are applied using a first liquid stream flow rate to second liquid stream flow rate ratio of from about 1 : 1 to about 500: 1. In some embodiments, the first liquid stream flow rate to second liquid stream flow rate ratio is from about 100:1 to about 500:1.474248642. vl5726.1016001

[0164] Applicant has surprisingly discovered that controlling the dehydration kinetics (e.g. , the flow rate ratio between the first liquid stream and second liquid stream, or the timing of dehydration) may influence particle size and particle properties (e.g., morphology). For example, faster flow rates alone or in combination with lowering the concentration of the therapeutic agent in the aqueous feed solution have been found to form smaller particles (e.g., a d50 less than about 10 pm on average). In some embodiments, the first liquid stream flow rate to second liquid stream flow rate ratio is about 1 : 10 to about 1 :250. In certain embodiments, the first liquid stream flow rate to second liquid stream flow rate ratio depends on the organic liquid within the second liquid stream. For example, where the organic liquid comprises an alkylacetate, Applicant has surprisingly discovered that the flow rate ratio of between about 1:10 to about 1:250 produces droplets and particles having particularly advantageous properties. In certain embodiments, an organic liquid that comprises an alkyl group produces advantageous properties with a flow rate ratio of between about 1 :5 to about 1 :20.

[0165] In some embodiments, dispersing a first liquid stream into a second liquid stream at or after a junction is performed at a temperature of from about 0°C to about 40°C (e.g., about 0°C to about 15°C, about 10°C to about 20°C, about 20°C to about 30°C, 30°C to about 40°C, about 15°C to about 30°C, about 18°C to about 30°C, about 18°C to about 25°C, etc.). In some embodiments, dispersing a first liquid stream into a second liquid stream at or after a junction is performed at a temperature of from about 18°C to about 22°C.

[0166] In some embodiments, in any of the methods described herein comprising the use of a first and / or second liquid stream, the method may further comprise providing one or more additional liquids (e.g., providing a liquid stream comprising the additional liquid) at or after the junction. The additional liquid stream (e.g., a second, third, fourth, fifth, etc. liquid stream) may comprise the same, or different, liquid (e.g., an organic liquid, an organic solvent, a dehydration liquid) as the first and / or second liquid stream. In some embodiments, the additional liquid stream is added at a second junction. In certain embodiments, a third liquid stream comprising an organic liquid is provided after the first junction, wherein the third liquid stream comprises the same organic liquid as the second liquid stream. In certain other embodiments, a third liquid stream comprising a dehydration liquid is provided after the first junction, wherein the third liquid stream comprises a different liquid to the second liquid stream. In some embodiments, the third liquid stream is added at a third junction.Formation of Particles484248642. vl5726.1016001

[0167] The particles as described herein, can be formed, for example, by dispersing a first liquid stream comprising an aqueous feed solution or aqueous liquid into a second liquid stream comprising an organic liquid, forming aqueous droplets (e.g., by subjecting the aqueous feed solution and organic liquid to turbulent flow conditions), and removing water from the aqueous droplets by maintaining the aqueous droplets in the presence of a dehydration liquid. In some embodiments, the dehydration liquid is the organic liquid. In some embodiments, the methods comprise the continuous formation of aqueous droplets in an organic liquid, and the processing of the droplets to produce particles. In certain embodiments, particle formation begins to take place when at least a subset of the components of the aqueous droplets begin to undergo dehydration, precipitation or phase separation as water is removed. In embodiments, the aqueous droplets are dried by maintaining the aqueous droplets comprising the therapeutic agent in continuous contact with an organic liquid.

[0168] In some embodiments, particles are formed after the aqueous feed solution or aqueous liquid disperses within the organic liquid, e.g., through a diffusion or dispersion process. In other embodiments the organic liquid may have varying degrees of miscibility with the aqueous liquid and represent a weakly or negligibly solubilizing medium in relation to the components of the particles or a subset of the components of the particles, e.g., the therapeutic agent. In some embodiments the aqueous liquid may be immiscible in the organic liquid. The therapeutic agent, e.g., antibody or antibody fragment, is typically less soluble in the organic liquid relative to the aqueous liquid in the timeframe of or under the conditions of production, e.g., at least about 5, 10, 100, or about 1000 times less soluble. In some embodiments, the organic liquid may be an organic solvent. The organic liquid can further include a carbohydrate, a pH adjusting agent, a salt, a chelator, a mineral, a polymer, a surfactant, an amino acid, an oligopeptide, a biologic excipient, a chemical excipient, an antiseptic, an antioxidant, a paraben, a bactericide, a fungicide, a vitamin, a preservative, an analgesic, a nutrient media, a flocculation agent, or a combination thereof. Example aqueous liquids may contain stabilizers, e.g., crowding agents. These solutions, in certain embodiments, include excipients such as a salt (e.g., sodium chloride), sugars and sugar alcohols (e.g., sorbitol, dextran 40, dextran 6000, or trehalose), polymers e.g., polyacrylamide, PEG 3350, PEG 300, PEG 8000, PEG 20k, Ficoll 400, Ficoll 70, or polyvinylpyrrolidone, e.g, Povidone), a protein (e.g, or human serum albumin (HSA)), or a combination thereof. In still other embodiments, where the aqueous liquid is water, particles are obtained via osmotic drying of the aqueous droplets. The organic liquid that is used to continuously dry the particles, in certain embodiments, includes a high494248642. vl5726.1016001concentration of a solute (therapeutic agent and / or excipient, e.g., surfactant), e.g., at least about 0.03 osmol, at least about 0.2 osmol, at least about 1.0 osmol, or at least about 1.2 osmol.

[0169] In some embodiments, the aqueous liquid and the organic liquid are immiscible. As the two liquids interact with each other in a fluid channel or fluid vessel, turbulent flow conditions may be applied within the fluid channel or fluid vessel. The immiscible organic liquid may apply a shearing force on the aqueous liquid causing aqueous droplets to form. In certain embodiments, the organic liquid and the organic solvent are the same. In certain embodiments, the organic liquid and the organic solvent are different. In certain embodiments, the organic liquid and the dehydration liquid are the same. In certain embodiments, the organic liquid and the dehydration liquid are different.

[0170] In some embodiments, the aqueous liquid and the organic liquid are partially miscible. As the two liquids interact with each other in a fluid channel or fluid vessel, turbulent flow conditions may be applied within the fluid channel or fluid vessel. The partially miscible organic liquid may apply a shearing force on the aqueous liquid in the fluid channel or fluid vessel causing aqueous droplets to form. At least a portion of an aqueous fraction in the aqueous droplets may be dissolved into the organic liquid, removing at least a portion of the aqueous fraction from the aqueous droplet. In certain embodiments, therapeutic in the aqueous liquid may not be miscible ( / .< ., therapeutic is immiscible) with the organic solvent and / or the dehydration liquid. As the portion of the aqueous liquid is dissolved ( / .< ., the fraction of the aqueous liquid which is miscible) therapeutic may remain. As a result, the aqueous droplets formed may be subjected to dehydration leaving therapeutic behind to form a particle including therapeutic. In certain embodiments, the organic liquid and the organic solvent are the same. In certain embodiments, the organic liquid and the organic solvent are different. In certain embodiments, the organic liquid and the dehydration liquid are the same. In certain embodiments, the organic liquid and the dehydration liquid are different. In certain embodiments, the organic solvent and the dehydration liquid are the same. In certain embodiments, the organic solvent and the dehydration liquid are different.

[0171] In other embodiments, the surfactant in the organic liquid helps to prevent coalescence of the aqueous droplets. In certain embodiments, an oligopeptide excipient, a protein excipient, the therapeutic agent(s) themselves, e.g, antibody, antibody fragment, or human serum albumin (HSA), or a combination thereof, act as surfactants.

[0172] In some embodiments, droplets (e.g., aqueous droplets) are dehydrated by an organic liquid, an organic solvent, a dehydration liquid, or a combination thereof. In other embodiments, dehydration of droplets (e.g., dehydrating an aqueous fraction of aqueous 504248642. vl5726.1016001droplets) comprises contacting the droplets with a dehydration liquid in tubing (e.g., a drying tube), a vessel (e.g., a drying vessel), or a combination thereof (e.g., continuous drying tube, a continuous drying vessel, or a combination thereof). In some embodiments, a drying tube is provided comprising a dehydration coil. In certain other embodiments, the drying tube is further connected to at least one process. In still other embodiments, the drying vessel is further connected to at least one process. The drying tube, drying vessel, or combination thereof may be connected to a process by a junction. In certain embodiments, the drying vessel comprises mechanical stirring. In certain embodiments, the mechanical stirring is conducted by a turbulent stirred vessel, a magnetic stirring device, or a mechanical stirring device. In some embodiments, the average residence time T of the particles in the drying vessel (e.g., continuous drying vessel) and the relationship between this time and the characteristic particle formation time t* is considered in the design and sizing of the drying vessel. In some embodiments, the average residence time can be estimated as T = V / Q, where V is the volume of the drying vessel and Q is the volumetric flow rate of product through it. In some embodiments, the particle resides in the drying vessel for about 0 to about 10,000 seconds, e.g., about 0 to about 5,000 seconds, 1,000 seconds, 500 seconds, 400 seconds, 300 seconds, 200 seconds, 100 seconds, 90 seconds, 80 seconds, 70 seconds, 60 seconds, 50 seconds, 40 seconds, 30 seconds, 20 seconds, 10 seconds, 9 seconds, 8 seconds, 7 seconds, 6 seconds, 5 seconds, 4 seconds, 3 seconds, 2 seconds, or 1 second. In some embodiments, the ratio of the characteristic particle formation time to the average residence time t* / z is a figure of merit. In some embodiments, the ratio of the characteristic particle formation time to the average residence time is about 0 to about 100, e.g., 0 to about 100, 50, 25, 10, 5, 4, 3, 2, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, or 0.1. In some embodiments, the particle resides in the drying vessel for at least about 1 minute, about 2 minutes, about 3 minutes, about 4 minutes, about 5 minutes, about 6 minutes, about 7 minutes, about 8 minutes, about 9 minutes, about 10 minutes, about 15 minutes, about 30 minutes, about 45 minutes, about 1 hour, about 2 hours, or about 3 or more hours.

[0173] In some embodiments, the residence time of droplets (e.g., aqueous droplets) in a tubing, a vessel, or a coil (e.g., a dehydration coil) is at least about 10 seconds (e.g., at least about 11 seconds, at least about 12 seconds, at least about 13 seconds, at least about 14 seconds, at least about 15 seconds, etc.). In some embodiments, the residence time of aqueous droplets in a drying tube or vessel is at least about 15 seconds. In some embodiments, the residence time of droplets in a coil is about 15 seconds. In certain embodiments, the tubing or coil has a defined length. In certain embodiments, the tubing or coil is between about 1 ft to about 50 ft long, e.g.,514248642. vl5726.1016001about 1 ft, about 2 ft, about 3 ft, about 4 ft, about 5 ft, about 6 ft, about 7 ft, about 8 ft, about 9 ft, about 10 ft, about 15 ft, about 20 ft, about 25 ft, about 30 ft, about 35 ft, about 40 ft, about 45 ft, or about 50 ft. In some embodiments, the tubing or coil may be between about 1 ft and about 50 ft, about 1 ft and about 5 ft, about 5 ft and about 10 ft, about 10 ft and about 15 ft, about 15 ft and about 20 ft, about 20 ft and about 25 ft, about 25 ft and about 30 ft, about 30 ft and about 35 ft, about 35 ft and about 40 ft, about 40 ft and about 45 ft, or about 45 ft and about 50 ft. In some embodiments, the tubing or coil may have an inner diameter of between about 0.1 in to about 3 in, e.g., about 0.1 in, 0.15 in, 0.2 in, 0.25 in, 0.3 in, 0.35 in, 0.4 in, 0.45 in, 0.5 in, 0.55 in, 0.6 in, 0.65 in, 0.7 in, 0.75 in, 0.8 in, 0.85 in, 0.9 in, 0.95 in, 1 in, 1.1 in, 1.15 in, 1.2 in, 1.25 in, 1.3 in, 1.35 in, 1.4 in, 1.45 in, 1.5 in, 1.55 in, 1.6 in, 1.65 in, 1.7 in, 1.75 in, 1.8 in, 1.85 in, 1.9 in, 1.95 in, 2 in, 2.1 in, 2.15 in, 2.2 in, 2.25 in, 2.3 in, 2.35 in, 2.4 in, 2.45 in, 2.5 in, 2.55 in, 2.6 in, 2.65 in, 2.7 in, 2.75 in, 2.8 in, 2.85 in, 2.9 in, 2.95 in, 3 in.

[0174] The actual continuous dehydration or desiccation time may vary as a function of the properties of the organic solvent in addition to changes that take place in the drop as the concentration of the solutes, precipitation of solutes, and / or phase separation begin to take place. Relevant properties of the organic solvent that influence the drying time include, e.g., the solubility of an aqueous fraction of the aqueous droplet in the organic solvent, the saturation state of the organic solvent, the diffusivity of the aqueous fraction of the aqueous droplet in the organic solvent, and the polarity of the organic solvent. The term “polarity” or “polarities” refer to the overall solvation capability (solvation power) of the aqueous fraction of the aqueous droplet, which in turn depends on the action of all possible, nonspecific and specific, intermolecular interactions between solute ions or molecules and solvent molecules, excluding, however, those interactions leading to definite chemical alterations of the ions of molecules of the solute (Chem. Rev., 1994, 94, 2319-2358). A prediction of liquid polarity may be made from their dielectric constant. Liquids with high dielectric constants are considered more polar and those with low dielectric constants are considered less polar or nonpolar (<~15). Depending on the chosen process conditions, dehydration of the aqueous fraction of the aqueous droplets to form particles may occur over a period of nanoseconds to days. In embodiments where the aqueous liquid is aqueous, drying times can vary, e.g., between 1 ps and 1000 s depending on the solvent chemistry. In some embodiments, the characteristic particle formation time can be estimated as: t* = pr2 / 3Dc (1 — / ?), where p is the density of the aqueous liquid, r is the initial radius of the droplet, D is the diffusivity of the aqueous liquid in the organic liquid, c is the solubility of the aqueous liquid in the organic liquid, and ft is the524248642. vl5726.1016001fractional level of saturation of the organic liquid with the aqueous liquid at the start of the particle formation process.

[0175] The term “primary dehydration” refers to a step by which a droplet comprising an aqueous liquid is maintained within an organic solvent and dehydrated or desiccated by the organic solvent, e.g., through dispersal of an aqueous fraction of the droplet in the organic solvent, to form a particle. In some embodiments, the methods described herein use a primary dehydration process that is continuous. The term “secondary dehydration” refers to a step of removing residual moisture and / or liquid from particles, e.g., modifying the residual moisture and / or aqueous liquid and organic liquid content of the particles. In some embodiments, the methods described herein use a secondary dehydration process that is continuous. Example methods of secondary dehydration include washing the particles with a dehydration liquid, vacuum drying, air filter drying, with or without the application of heat, lyophilization, fluidized bed drying, tray drying, belt drying, or slurry spray drying. In some embodiments, secondary dehydration comprises sparging. In certain embodiments, aqueous liquid and / or organic liquid is removed through centrifugation, sieving, filtration, settling, magnetic collection, solvent exchange, decanting, hydrocyclone separation, or a combination thereof. In certain embodiments, the filtration is cross-flow filtration, tangential flow filtration or normal flow filtration. In some embodiments, the methods described herein comprise more than one secondary dehydration step (e.g., solvent exchange and sparging).

[0176] In certain embodiments of the continuous dehydrating process, the organic liquid includes or is in contact with a drying substance, / .<?., a desiccant, to absorb the aqueous liquid or otherwise sequester it, e.g., by reaction. Such substances can be useful for ensuring a uniform, steady-state degree of saturation of the aqueous liquid in the organic liquid during the continuous dehydrating process. Example desiccants include, but are not limited to celite, molecular sieves, phosphorous pentoxide, magnesium sulfate, silica, calcium chloride, activated charcoal, potassium carbonate, activated alumina, or a combination thereof.Particles

[0177] In some embodiments, any of the methods and processes described herein produce a collection of particles having uniform or nonuniform properties or morphology. In some embodiments, a portion of the particles have no visible internal void spaces (when assessed through FIB-SEM). In some embodiments, a portion of the particles have less than about 90% internal void spaces, e.g., less than about 90, 85, 80, 75, 70, 65, 60, 55, 50, 45, 40, 35, 30, 25, 24, 23, 22, 21, 20, 15, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, or 0.5% internal void spaces (e.g., after534248642. vl5726.1016001removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In certain embodiments, the particles have less than about 90% internal void spaces, less than about 80% internal void spaces, less than about 70% internal void spaces, less than about 60% internal void spaces, less than about 50% internal void spaces, less than about 40% internal void spaces, less than about 30% internal void spaces, less than about 25% internal void spaces, less than about 20% internal void spaces, less than about 10% internal void spaces, less than about 5% internal void spaces, less than about 1% internal void spaces, or less than about 0.5 % internal void spaces (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In embodiments, the particles are substantially free from any internal void spaces (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent).

[0178] Some embodiments of this disclosure may include a population of particles in a suspension having varying void space. By way of example, an embodiment of this disclosure may include a population of particles including a portion of the particles having approximately 90% internal void space, a portion of the particles having approximately 20% internal void space, and a portion of the particles having approximately 3% internal void space. The population of particles in embodiments of this disclosure may have any combination of particles having varying amounts of internal void space as disclosed above without departing from the scope of this disclosure.

[0179] Suitable methods for determining internal void space can be performed using Focused Ion Beam Scanning Electron Microscopy (FIB-SEM), which can be used to visualize “accessible” and “inaccessible” void spaces, or gas displacement pycnometry (Micromeritics Instrument Corporation of Norcross, Ga), which can determine “accessible” voids (void spaces accessible from the surface rather than those resembling a core-shell structure that are “unaccessible form the surface”). Gas pycnometry is a common analytical technique that uses a gas displacement method to measure volume. Inert gases, such as helium or nitrogen, are used as the displacement medium. True volume is total volume minus volume accessible to the gas. Density is calculated by dividing sample weight with true volume. The sample is sealed in the instrument compartment of a known volume, the appropriate inert gas is admitted, and then expanded into another precision internal volume. The pressure before and after expansion is measured and used to compute the sample volume. Dividing this volume into the sample weight gives the gas displacement density. Cross-sections of typical particles of the present disclosure 544248642. vl5726.1016001indicate an absence of pores (substantially free from any internal void spaces) and low particle porosity as shown by FIB-SEM or by gas pycnometry using helium at temperatures at about 22 °C to provide densities typically averaging about 1.3 g / cm3with standard deviations at about 0.0005 g / cm3. For example, internal void space can be calculated using the following formula: internal void space = AV / AP, where Avis the total area of void spaces and APis the total area of the particle.

[0180] In other embodiments, at least a portion of the population of particles (e.g., a plurality of particles) may exhibit a porosity from about 0 to about 50% (e.g., from about 0 to about 10%, from about 0 to about 5%, from about 0 to about 1%, from about 0 to about 0.5%, from about 0 to about 0.1%, or from about 0 to about 0.01%). In some embodiments, a portion of the particles may exhibit a porosity from about 0 to about 50% (e.g., from about 0 to about 10%, from about 0 to about 5%, from about 0 to about 1%, from about 0 to about 0.5%, from about 0 to about 0.1%, or from about 0 to about 0.01%) (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In some embodiments, a portion of the particles may exhibit less than about 50% porosity (e.g., less than about 50%, less than about 40%, less than about 30%, less than about 20%, less than about 10%, less than about 5%, less than about 4%, less than about 4%, less than about 2%, less than about 1%, less than about 0.1%, less than about 0.01%).

[0181] Example pore size measurements include scanning electron microscopy (SEM), transmission electron microscopy (TEM), and confocal laser scanning microscopy analysis. A gallium focused ion beam (FIB) was used to cut one of the particles in half to reveal a crosssection of the particle interior. The specific surface area of porous micro- and nanospheres may also be investigated by nitrogen adsorption / desorption analysis and a Brunauer-Emmett-Teller adsorption model. In certain embodiments where the pore sizes are sufficiently large, mercury -intrusion porosimetry may be employed.

[0182] In some embodiments, a population of particles is provided, wherein a portion of the particles are circular. Circularity can serve as an indicator of the shape of the particle. The particles described herein, can have a characteristic circularity, e.g, have a relative shape, that is substantially circular. This characteristic describes and defines the form of a particle on the basis of its circularity. The circularity is 1.0 when the particle has a completely circular structure. At least a portion of the population of particles as described herein, have a circularity of at least about 0.8, 0.9, 0.95, 0.96, 0.97, 0.98, or 0.99 (e.g, after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a 554248642. vl5726.1016001therapeutic agent); greater than about 0.80, greater than about 0.90, greater than about 0.95, or greater than about 0.98 (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In some embodiments, the circularity of the portion of the particles is greater than about 0.88 (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In other embodiments, the circularity of the portion of the particles is greater than about 0.90 (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In certain embodiments, the circularity of the portion of the particles is greater than about 0.93 (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In embodiments, the circularity of the portion of the particles is greater than about 0.97 (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). The diameter and the circularity of the portion of the population of particles can be determined by the image processing of an image observed under an electron microscope or the like or a flow-type particle image analyzer. The circularity can also be determined by subjecting a portion of the particles to circularity measurement and averaging the resulting values. For example, circularity (circ) can be calculated using the following formula:

[0183] The term “perimeter”, as used herein, refers to the boundary of a closed plane figure or the sum of all borders of a two-dimensional image. As used herein, the term “area”, refers to the cross-sectional area of a two-dimensional image of at least one particle. The circularity of a particle can also be described as the ratio of the smallest dimension of the particle to its largest diameter. For a perfect circle, the ratio is 1. The percentage circularity can be calculated by multiplying the circularity by 100. The circularity can be calculated, for example, by measuring the aspect ratio using any software adapted to deal with images, for example, images obtained by microscopy, in particular, scanning electron microscopy (SEM) or transmission electron microscopy (TEM). In some embodiments, the circularity of the portion of the particles is at least about 10% (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent), e.g., at least about 20, 30, 40, 50, 55, 60, 65, 70, 75, 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or about 100% (e.g., after removing an aqueous fraction from a564248642. vl5726.1016001droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In other embodiments, the circularity of the particles is at least about 88% (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In certain embodiments, the circularity of the particles is at least about 90% (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In still other embodiments, the circularity of the particles is at least about 93% (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In embodiments, the circularity of the particles is at least about 97% (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent).

[0184] In some embodiments, the circularity (e.g., number- weighted average circularity) of a portion (e.g., more than one (1), about two (2), about a quarter, about a half, about three-quarters, about a plurality, etc.) of the particles is from about 0.10 to about 1.00 (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent), e.g., at least from about 0.20, 0.30, 0.40, 0.50. 0.60, 0.70, 0.75, 0.80, 0.81, 0.82, 0.83, 0.84, 0.85, 0.86, 0.87, 0.88, 0.89, 0.90, 0.91, 0.92, 0.93, 0.94, 0.95, 0.96, 0.97, 0.98, or 0.99 to about 1.00 (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In certain embodiments, the circularity of the portion of the particle is at least from about 0.80 to about 1.00 (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In certain embodiments, the circularity of the portion of the particle is at least from about 0.88 to about 1.00 (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In still other embodiments, the circularity of the portion of the particles is at least from about 0.90 to about 1.00 (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In certain other embodiments, the circularity of the portion of the particles is at least from about 0.93 to about 1.00 (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In embodiments, the circularity of the portion of the particles is at least from about 0.97 to about 1.00 (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream 574248642. vl5726.1016001to form particles comprising a therapeutic agent). In other embodiments, methods of measuring particle circularity include image analysis of scanning electron micrographs of the particles in which the average roundness is calculated on the basis of the cross-sectional shapes of a portion of the population of particles projected onto the plane of the image. Such roundness factors can be extended to identify the corresponding circularity.

[0185] In other embodiments of the present disclosure, the dehydration or drying operation may be controlled to provide a portion of particles in a population of particles having particular characteristics, such as particles having a substantially smooth surface. “Surface roughness”, as used herein, means a particle having numerous wrinkles or creases, e.g., being ridged or wrinkled. The term “pit”, as used herein, refers to an indentation or crevice in the particle, either an indentation or crevice in the two-dimensional image or an indentation or crevice in an object. The term “spike”, as used herein, refers to a projection pointing outward from the centroid of a particle, a projection pointing outward from the centroid of a two-dimensional image or a sharp projection pointing outward from an object. As described herein, the dehydration process may be controlled by adjusting the contact time between aqueous droplets and a dehydration liquid. The dehydration liquid may be chosen, as disclosed herein, to control the Peclet number or the rate of dehydration of the aqueous droplets to form a portion of particles in the population in the suspension without pits, spikes, wrinkles, creases, or other forms of surface roughness.

[0186] In embodiments of the present disclosure, a portion of particles in a population of particles as described herein, have a surface morphology that is smooth rather than ridged or wrinkled. The surface roughness of the particles may be decreased by controlling the formulation and / or process to form the particles as described herein. In certain embodiments, the drying conditions can be selected to control the particle morphology in order to enhance the smoothness of the particle’s surface. In particular, the drying conditions can be selected to provide a portion of particles in a population having a substantially smooth surface. In certain embodiments, the portion of particles in the population have a substantially smooth surface (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). A person of ordinary skill in the field of this disclosure can readily assess the surface morphology of the disclosed particles using routine and standard techniques.

[0187] In some embodiments, a portion of particles in a population have a volume-weighted average diameter of at least about 0.01 pm to about 1000 pm (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form 584248642. vl5726.1016001particles comprising a therapeutic agent), e.g., at least about 0.01 to about 900, 800, 700, 600, 500, 400, 300, 200, 100, 90, 80, 70, 60, 50, 45, 40, 35, 30, 25, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.09, 0.08, 0.07, 0.06, 0.05, 0.04, 0.03, 0.02, or about 0.01 pm (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In certain embodiments, a portion of the particles have a volume-weighted average diameter of at least about 1 pm to about 100 pm (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent), e.g., at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, or 50 to about 100 pm (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In still other embodiments, at least a portion of the particles have a volume-weighted average diameter of at least about 4 pm to about 100 pm (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In certain other embodiments, a portion of the particles have a volume-weighted average diameter of at least about 10 pm to about 100 pm (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In some embodiments, a portion of the particles have a volume-weighted average diameter of at least about 10 pm to about 95 pm (e.g. , after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In some embodiments, a portion of the particles have a volume-weighted average diameter of at least about 10 pm to about 75 pm (e.g. , after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In some embodiments, a portion of the particles have a volume-weighted average diameter of at least about 10 pm to about 50 pm (e.g. , after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In certain embodiments, a portion of the particles in a population are intentionally controlled in their diameter. In other embodiments, a portion of the particles have volume-weighted average diameters from at least about 0.1 pm to about 1000 pm (e.g. , after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent), e.g., at least about 1 pm to about 400 pm, at least about 1 pm to about 200 pm, at least about 1 pm to about 100 pm, at least about 1 pm to about 50 pm, at least about 1 pm to about 25 pm, at least about 1 pm to about 10 pm, at least about 10 pm to about 100 pm,594248642. vl5726.1016001at least about 50 pm to about 100 pm, at least about 50 pm to about 75 pm, or at least about 75 pm to about 100 pm (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In some embodiments, a portion of the particles have volume-weighted average diameters from at least about 1 pm to about 100 pm (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent), e.g., from at least about 4 pm to about 100 pm, from at least about 10 pm to about 100 pm, or from at least about 20 pm to about 50 pm (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent).

[0188] In certain embodiments, a portion of the particles have a volume-weighted average diameter of at least about 0.1 pm to about 1000 pm (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In certain other embodiments, a portion of the particles have a volume-weighted average diameter of at least about 1 pm to about 100 pm (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In still other embodiments, a portion of the particles have a volume-weighted average diameter of at least about 5 pm to about 100 pm (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In certain embodiments, a portion of the particles have a volume-weighted average diameter of at least about 5 pm to about 50 pm (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In embodiments, a portion of the particles have a volume-weighted average diameter of at least about 5 pm to about 20 pm (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent).

[0189] In some embodiments, a suspension comprises a population of particles wherein at least a plurality of particles have an volume-weighted average diameter of between at least about 0.1 pm to about 1000 pm (e.g., at least about 1 pm to about 400 pm, at least about 1 pm to about 200 pm, at least about 1 pm to about 100 pm, at least about 1 pm to about 50 pm, at least about 1 pm to about 25 pm, at least about 1 pm to about 10 pm, at least about 10 pm to about 100 pm, at least about 50 pm to about 100 pm, at least about 50 pm to about 75 pm, at least or about 75 pm to about 100 pm). In certain embodiments, a suspension comprises a population of particles wherein at least a plurality of particles have a volume-weighted average 604248642. vl5726.1016001diameter of between at least about 1 to about 100 pm (e.g., at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, or 50 to about 100 pm). In still other embodiments, a suspension comprises a population of particles wherein at least a plurality of particles have a volume-weighted average diameter of between at least about 4 pm to about 100 pm. In certain other embodiments, a suspension comprises a population of particles wherein at least a plurality of particles have a volume-weighted average diameter of between at least about 10 pm to about 100 pm. In embodiments, a suspension comprises a population of particles wherein at least a plurality of particles have a volume-weighted average diameter of between at least about 20 pm to about 50 pm. In some embodiments, a suspension comprises a population of particles wherein at least a plurality of particles have a volume-weighted average diameter of between at least about 1 pm to about 100 pm. In some embodiments, a suspension comprises a population of particles wherein at least a plurality of particles have a volume-weighted average diameter of between at least about 5 pm and about 50 pm. In some embodiments, a suspension comprises a population of particles wherein at least a plurality of particles have a volume-weighted average diameter of between at least about 10 pm and about 20 pm. In certain embodiments, the suspension comprises a population of particles wherein at least a plurality of particles have a volume-weighted average diameter that is equal to or smaller than l / 5ththe inner diameter of a syringe needle for injecting the suspension. In some embodiments, a volume-weighted average diameter of the plurality of particles to the inner diameter of a syringe needle for administering the suspension may be 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20, etc.

[0190] In some embodiments, the diameter of the particles produced according to the disclosure may be controlled by affecting the amount of turbulence or shearing force applied to an aqueous feed solution in an organic liquid. In certain embodiments, the turbulence introduced to the system may apply a shearing force to the aqueous feed solution to form the aqueous droplets as disclosed herein. According to this disclosure, factors that may be controlled to affect the formation of aqueous droplets may include applying turbulent conditions or highly turbulent conditions; the flow rate of an aqueous feed solution; the flow rate of an organic liquid; the relative flow rates between the aqueous feed solution and the organic liquid; the viscosity of the aqueous feed solution; the viscosity of the organic liquid; the relative viscosity of the aqueous feed solution to the viscosity of the organic liquid; the introduction of shearing forces in a fluid channel or a fluid vessel by a fitting, rotor stator mixer, Tee-mixer, or static mixer at a junction; changing an internal diameter of at least one fluid channel at one or more junctions; or applying an increase in / decrease in / or maintained pressure 614248642. vl5726.1016001to the at least one fluid channel to affect a flow rate of at least one liquid in the at least one fluid channel.

[0191] In other embodiments, a portion of particles in a population exhibit a volume-weighted average skeletal density from at least about 1.00 to about 6.00 g / cm3(e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent), e.g., from at least about 1.00 to about 5.00 g / cm3, from at least about 1.00 to about 3.00 g / cm3, from at least about 1.00 to about 2.00 g / cm3, from at least about 1.00 to about 1.50 g / cm3, from at least about 1.30 to about 1.50 g / cm3, from at least about 1.30 to about 1.50 g / cm3, or from at least about 1.10 to about 1.40 g / cm3(e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In some embodiments, a portion of the particles in a population exhibit a volume-weighted average skeletal density from at least about 0.10 to about 5.00 g / cm3(e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent), e.g., from at least about 0.10 to about 2.50 g / cm3, from at least about 0.10 to about 1.40 g / cm3, from at least about 0.50 to about 1.40 g / cm3, or from at least about 1.00 to about 1.40 g / cm3(e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In certain embodiments, a portion of the particles in a population have a volume-weighted average skeletal density of at least about 0.90 to about 1.60 g / cm3(e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In still other embodiments, a portion of the particles in a population have a volume-weighted average skeletal density of at least about 1.30 to about 1.58 g / cm3(e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In embodiments, at least a portion of the particle in a population have a volume-weighted average skeletal density of at least about 1.30 to about 1.50 g / cm3(e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). The terms “skeletal density”, “envelope density” and “density” may be used interchangeably. The terms “skeletal density”, “envelope density” or “density” as used throughout the specification, examples, and claims is intended to show pore volume and internal void space of the particles. Example methods of skeletal density measurements include gas displacement pycnometry and / or mercury intrusion.624248642. vl5726.1016001

[0192] In certain embodiments, a portion of the particles in a population have a volume-weighted average skeletal density of at least about 1000 mg / mL to about 1500 mg / mL, at least about 1050 mg / mL to about 1500 mg / mL, at least about 1100 mg / mL to about 1500 mg / mL, at least about 1150 mg / mL to about 1500 mg / mL, at least about 1200 mg / mL to about 1500 mg / mL, at least about 1250 mg / mL to about 1500 mg / mL, at least about 1300 mg / mL to about 1500 mg / mL, at least about 1310 mg / mL to about 1500 mg / mL, at least about 1320 mg / mL to about 1500 mg / mL, at least about 1330 mg / mL to about 1500 mg / mL, at least about 1340 mg / mL to about 1500 mg / mL, at least about 1350 mg / mL to about 1500 mg / mL, at least about 1360 mg / mL to about 1500 mg / mL, at least about 1370 mg / mL to about 1500 mg / mL, at least about 1380 mg / mL to about 1500 mg / mL, at least about 1390 mg / mL to about 1500 mg / mL, at least about 1400 mg / mL to about 1500 mg / mL, at least about 1410 mg / mL to about 1500 mg / mL, at least about 1420 mg / mL to about 1500 mg / mL, at least about 1430 mg / mL to about 1500 mg / mL, at least about 1440 mg / mL to about 1500 mg / mL, at least about 1450 mg / mL to about 1500 mg / mL, at least about 1460 mg / mL to about 1500 mg / mL, at least about 1470 mg / mL to about 1500 mg / mL, at least about 1480 mg / mL to about 1500 mg / mL, at least about 1490 mg / mL to about 1500 mg / mL, or at least about 1200 mg / mL to about 1350 mg / mL (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In certain other embodiments, a portion of the particles in a population have a volume-weighted average skeletal density of greater than at least about 1000 mg / mL, at least about 1050 mg / mL, at least about 1100 mg / mL, at least about 1150 mg / mL, at least about 1200 mg / mL, at least about 1250 mg / mL, at least about 1300 mg / mL, at least about 1310 mg / mL, at least about 1320 mg / mL, at least about 1330 mg / mL, at least about 1340 mg / mL, at least about 1350 mg / mL, at least about 1360 mg / mL, at least about 1370 mg / mL, at least about 1380 mg / mL, at least about 1390 mg / mL, at least about 1400 mg / mL, at least about 1410 mg / mL, at least about 1420 mg / mL, at least about 1430 mg / mL, at least about 1440 mg / mL, at least about 1450 mg / mL, at least about 1460 mg / mL, at least about 1470 mg / mL, at least about 1480 mg / mL, at least about 1490 mg / mL, or at least about 1500 mg / mL (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent).

[0193] In other embodiments, a portion of the particles in a population exhibit a volume-weighted average therapeutic density from at least about 1.00 to about 6.00 g / cm3(e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent), e.g., from at least about 1.00 to about 5.00 g / cm3, from at least about 1.00 to about 3.00 g / cm3, from at least about 1.00 to about 2.00634248642. vl5726.1016001g / cm3, from at least about 1.00 to about 1.50 g / cm3, from at least about 1.30 to about 1.50 g / cm3, from at least about 1.30 to about 1.50 g / cm3, or from at least about 1.10 to about 1.40 g / cm3(e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In some embodiments, a portion of the particles in a population exhibit a volume-weighted average therapeutic density from at least about 0.10 to about 5.00 g / cm3(e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent), e.g., from at least about 0.10 to about 2.50 g / cm3, from at least about 0.10 to about 1.40 g / cm3, from at least about 0.50 to about 1.40 g / cm3, or from at least about 1.00 to about 1.40 g / cm3(e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In certain embodiments, a portion of the particles in a population have a volume-weighted average therapeutic density of at least about 0.90 to about 1.60 g / cm3(e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In still other embodiments, a portion of the particles have a volume-weighted average therapeutic density of at least about 1.30 to about 1.58 g / cm3(e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In embodiments, a portion of the particles have a volume-weighted average therapeutic density of at least about 1.30 to about 1.50 g / cm3(e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). The terms “therapeutic density” and “therapeutic agent density” may be used interchangeably. The terms “therapeutic density” and “therapeutic agent density” as used throughout the specification, examples, and claims is intended to describe a therapeutic loading in the particles. Example methods of therapeutic density measurements include gas displacement pycnometry and / or mercury intrusion.

[0194] In some embodiments, a portion of the particles (e.g., solid particles) have a volume-weighted average therapeutic agent density of at least about 0.1% by mass to about 100% by mass (e.g., at least about 0.1% by mass to about 100%, about 99.9%, about 99%, about 98%, about 97%, about 96%, about 95%, about 94%, about 93%, about 92%, about 91%, about 90%, about 85%, about 80%, about 75%, about 70%, about 65%, about 60%, about 55%, about 50%, about 45%, about 40%, about 35%, about 30%, about 25%, about 20%, about 15%, about 10%, about 9%, about 8%, about 7%, about 6%, about 5%, about 4%, about 3%, about 2%, about 1%). In certain embodiments, a portion of the particles in a population have a 644248642. vl5726.1016001volume-weighted average therapeutic agent density of at least about 0.1% by mass (e.g., at least about 0.1% by mass, about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, about 99.9%, about 100%).

[0195] In some embodiments, a portion of the particles (e.g., solid particles) have a volume-weighted average therapeutic agent density of at least about 0.1% by volume to about 100% by volume (e.g., at least about 0.1% by volume to about 100%, about 99.9%, about 99%, about 98%, about 97%, about 96%, about 95%, about 94%, about 93%, about 92%, about 91%, about 90%, about 85%, about 80%, about 75%, about 70%, about 65%, about 60%, about 55%, about 50%, about 45%, about 40%, about 35%, about 30%, about 25%, about 20%, about 15%, about 10%, about 9%, about 8%, about 7%, about 6%, about 5%, about 4%, about 3%, about 2%, about 1%). In certain embodiments, a portion of the particles in a population have a volume-weighted average therapeutic agent density of at least about 0.1% by volume (e.g., at least about 0.1% by volume, about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, about 99.9%, about 100%).

[0196] In some embodiments, a portion of the particles in a population can be characterized by a glass transition temperature of about 0 °C to about 250 °C (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent), e.g., of about 34 °C to about 200 ° C, of about 60 °C to about 170 °C, of about 90 °C to about 170 °C, of about 100 to about 170 °C, of about 130 to about 170 °C, of about 150 to about 170 °C, or of about 160 to about 170 °C (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). The term “glass transition” as used herein, refers to a thermodynamic transition of an amorphous material characterized by step changes in specific heat capacity and modulus. In embodiments, a portion of the particles that are produced by the methods described herein are amorphous. At temperatures above the glass transition temperature, molecular mobility is increased as are the rates of physical and chemical changes of the therapeutic agent. Example analytical methods for the determination of the glass transition temperature include differential scanning calorimetry and dynamic mobility analysis.654248642. vl5726.1016001In other embodiments, a portion of the particles in a population have a glass transition temperature of about 60 to about 170 °C (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In still other embodiments, a portion of the particles in a population have a glass transition temperature of about 60 to about 100 °C (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In certain embodiments, a portion of the particles in a population have a glass transition temperature of about 75 to about 80 °C (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In some embodiments, a portion of the particles in a population have a glass transition temperature of about 60 to about 120 °C (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent).

[0197] In certain embodiments, a portion of the particles in a population have a glass transition temperature that is higher than about 50 °C (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In certain other embodiments, a portion of the particles in a population have a glass transition temperature that is higher than about 90 °C (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In still other embodiments, a portion of the particles in a population have a glass transition temperature that is higher than about 100 °C (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In certain embodiments, a portion of the particles in a population have a glass transition temperature that is higher than about 160 °C (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In embodiments, a portion of the particles in a population have a glass transition temperature that is higher than about 170 °C (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In some embodiments, a portion of the particles in a population have a glass transition temperature of about 60 to about 100 °C (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In other embodiments, a portion of the particles in a population have a glass transition temperature of about 75 to about664248642. vl5726.101600180 °C (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent).

[0198] In some embodiments, a portion of the particles in a population further comprise a carbohydrate, a pH adjusting agent, a salt, a chelator, a mineral, a polymer, a protein stabilizer, an emulsifier, an antiseptic, an amino acid, an antioxidant, a protein, an organic solvent, a paraben, a bactericide, a fungicide, a vitamin, a preservative, a nutrient media, an oligopeptide, a biologic excipient, a chemical excipient, a surfactant, or a combination thereof. In some embodiments, the portion of the particles in the population further comprise one or more excipients (e.g., two or more excipients). In some embodiments, an excipient comprises a surfactant, an amino acid, a carbohydrate, a salt, an antioxidant, or a combination thereof. In certain embodiments, the excipients within a particle comprise the same excipients within the aqueous feed solution used to form the particle.

[0199] In other embodiments, the carbohydrate may be from the families of monosaccharides, disaccharides, oligosaccharides, or polysaccharides. In some embodiments, the carbohydrate is dextran, trehalose, sucrose, agarose, mannitol, lactose, sorbitol, maltose, starch, alginates, xanthan, galactomanin, agar, agarose, or a combination thereof. In certain embodiments, the carbohydrate is dextran, trehalose, sucrose, agarose, mannitol, lactose, sorbitol, maltose, hydroxypropyl beta-cyclodextrin, sulfobutylether beta-cyclodextrin, or a combination thereof. In embodiments, the carbohydrate is trehalose, cyclodextrins, hydroxypropyl beta-cyclodextrin, sulfobutylether beta-cyclodextrin, sucrose, or a combination thereof. In certain embodiments, the carbohydrate is trehalose. Cyclodextrins are available in three different forms a, P, and y based on the number of glucose monomers. The number of glucose monomers in a, P, and y cyclodextrin can be 6, 7, or 8, respectively.

[0200] In some embodiments, the pH adjusting agent is acetate, citrate, glutamate, glycinate, histidine, lactate, maleate, phosphate, succinate, tartrate, bicarbonate, aluminum hydroxide, phosphoric acid, hydrochloric acid, DL-lactic / glycolic acids, phosphorylethanolamine, tromethamine, imidazole, glyclyglycine, monosodium glutamate, sodium hydroxide, potassium hydroxide, or a combination thereof. In other embodiments, the pH adjusting agent is citrate, histidine, phosphate, succinate, sodium hydroxide, potassium hydroxide, or a combination thereof. In certain embodiments, the pH adjusting agent is citrate, histidine, phosphate, succinate, or a combination thereof. In certain other embodiments, the pH is about 4.5 to about 7.5. In embodiments, the pH is about 5.5 to about 6.5.

[0201] In certain embodiments, the charge of a portion of the particles in a population in a suspension may be adjusted to repel at least a second portion of the particles in the population 674248642. vl5726.1016001in the suspension. The portion of the particles may be charged by adjusting the pH of the aqueous feed solution or the portion of the particles with a pH adjusting agent. The respective portions of the particles in the population in the suspension may repel each other to suspend the particles and reduce sedimentation and / or settling within the suspension. In some embodiments, a plurality of the particles in population in the suspension are charged to repel each other within the suspension.

[0202] In other embodiments, the salt is sodium chloride, calcium chloride, potassium chloride, sodium hydroxide, stannous chloride, magnesium sulfate, sodium glucoheptonate, sodium pertechnetate, guanidine hydrochloride, potassium hydroxide, magnesium chloride, potassium nitrate, or a combination thereof. In embodiments, the salt is sodium chloride.

[0203] In some embodiments, the chelator is disodium edetate, ethylenediaminetetraacetic acid, pentetic acid, or a combination thereof. In other embodiments, the mineral is calcium, zinc, titanium dioxide, or a combination thereof. In certain embodiments, the polymer is glucose star polymer, silicone polymer, polydimethylsiloxane, polyethylene glycol (PEG), carboxymethylcellulose, poly(glycolic acid), poly(lactic-co-glycolic acid), polylactic acid, polycaprolactone (PCL), polyvinylpyrrolidone (PVP), ficoll, dextran, or a combination thereof.

[0204] In other embodiments, the protein stabilizer is acetyltryptophanate, caprylate, N-acetyltryptophan, trehalose, polyethylene glycol (PEG), polyoxamers, polyvinylpyrrolidone, polyacrylic acids, poly(vinyl) polymers, polyesters, polyaldehydes, tert-polymers, polyamino acids, hydroxy ethyl starch, N-methyl-2-pyrrolidone, sorbitol, sucrose, mannitol, or a combination thereof. In certain embodiments, the protein stabilizer is trehalose, polyethylene glycol (PEG), polyoxamers, polyvinylpyrrolidone, polyacrylic acids, poly(vinyl) polymers, polyesters, polyaldehydes, tert-polymers, polyamino acids, hydroxy ethyl starch, N-methyl-2-pyrrolidone, sorbitol, sucrose, mannitol, cyclodextrin, hydroxypropyl beta-cyclodextrin, sulfobutylether beta-cyclodextrin, pentetic acid, or a combination thereof. In embodiments, the protein stabilizer is trehalose, cyclodextrin, hydroxypropyl beta-cyclodextrin, sulfobutylether beta-cyclodextrin, pentetic acid, or a combination thereof. In certain embodiments, the PEG is PEG 200, PEG 300, PEG 3350, PEG 8000, PEG 10000, PEG 20000, or a combination thereof. The stabilizers, used synonymously with the term “stabilizing agent”, as described herein, can be a salt, a carbohydrate, saccharides or amino acids, preferably a carbohydrate or saccharide admitted by the authorities as a suitable additive or excipient in pharmaceutical compositions. The term “stabilizer” refers to an excipient or a mixture of excipients which stabilizes the physical and / or chemical properties of a therapeutic agent, e.g., an antibody. In some embodiments, stabilizers prevent, e.g., degradation of the therapeutic agent during droplet 684248642. vl5726.1016001formation, desiccation, and / or storage of the particulate matter. Example stabilizers include, but are not limited to, sugars, salts, hydrophobic salts, detergents, reducing agents, cyclodextrins, polyols, carboxylic acids, and amino acids. A “stable” formulation as described herein, refers to a formulation in which the therapeutic agent retains an acceptable portion of its essential physical, chemical, or biological properties over an acceptable period of time. In the case of proteins, e.g., example methods of 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 certain embodiments, chemical stability of a protein is assessed by measuring the size distribution of the sample at several stages. These include, e.g., before particle formation (assessment of the feed solution), immediately after particle formation, and again after a period of storage, where storage takes place either within or in the absence of a suspension formulation carrier medium. In certain other embodiments, the size distribution is assessed by size exclusion chromatography (SEC-HPLC). The term “excipient” refers to an additive to a preparation or formulation, which may be useful in achieving a desired modification to the characteristics of the preparation or formulation. Such modifications include, but are not limited to, physical stability, chemical stability, and therapeutic efficacy. Example excipients include, but are not limited to a carbohydrate, a pH adjusting agent, a salt, a chelator, a mineral, a polymer, a surfactant, an amino acid, an oligopeptide, a biologic excipient, a chemical excipient, an antiseptic, an antioxidant, a paraben, a bactericide, a fungicide, a vitamin, a preservative, an analgesic, and / or nutrient media.

[0205] Examples of emulsifiers suitable for use in the particles include, but are not limited to, lipophilic agents having an HLB of less than 15, such as mixed fatty acid monoglycerides; mixed fatty acid diglycerides; mixtures of fatty acid mono- and diglycerides; lipophilic polyglycerol esters; glycerol esters including glyceryl monooleate, glyceryl dioleate, glyceryl monostearate, glyceryl distearate, glyceryl monopalmitate, and glyceryl dipalmitate; glyceryllacto esters of fatty acids; propylene glycol esters including propylene glycol monopalmitate, propylene glycol monostearate, and propylene glycol monooleate; sorbitan ester including sorbitan monostearate, sorbitan sesquioleate; fatty acids and their soaps including stearic acid, palmitic acid, and oleic acid; and mixtures thereof glyceryl monooleate, glyceryl dioleate, glyceryl monostearate, glyceryl di stearate, glyceryl monopalmitate, and glyceryl dipalmitate; glyceryl-lacto esters of fatty acids; propylene glycol esters including propylene glycol monopalmitate, propylene glycol monostearate, and propylene glycol monooleate; sorbitan ester including sorbitan monostearate, sorbitan sesquioleate; fatty acids and their soaps 694248642. vl5726.1016001including stearic acid, palmitic acid, and oleic acid; phospholipids; or a combination thereof. In some embodiments, the emulsifier is polysorbate 80, polysorbate 60, polysorbate 20, e.g., Tween 80, Tween 60, Tween 20, sorbitan monooleate, ethanolamine, polyoxyl 35 castor oil, poloxyl 40 hydrogenated castor oil, carbomer 1342, a com oil-mono-di -triglyceride, a polyoxyethylated oleic glyceride, a poloxamer, or a combination thereof. In embodiments, the fatty acid ester of sorbitol is a sorbitan ester, e.g., span 20, span 40, span 60, or span 80. In certain embodiments, the emulsifier is polysorbate 80, sorbitan monooleate, or a combination thereof. In other embodiments, the antiseptic is phenol, m-cresol, benzyl alcohol, 2-phenyloxyethanol, chlorobutanol, neomycin, benzethonium chloride, gluteraldehyde, beta-propiolactone, or a combination thereof.

[0206] In certain embodiments, the amino acid is alanine, aspartic acid, cysteine, isoleucine, glutamic acid, leucine, phenylalanine, pyrrolysine, serine, selenocysteine, threonine, tryptophan, tyrosine, valine, asparagine, arginine, histidine, glycine, glutamine, proline, methionine, or various salts thereof (arginine hydrochloride, arginine glutamate, and the like) or a combination thereof. In certain other embodiments, the amino acid is alanine, aspartic acid, cysteine, isoleucine, glutamic acid, leucine, phenylalanine, pyrrolysine, serine, selenocysteine, threonine, tryptophan, tyrosine, valine, asparagine, arginine, histidine, glycine, glutamine, proline, methionine, or a combination thereof. In certain embodiments, the amino acid is arginine, histidine, proline, asparagine, or a combination thereof. In embodiments, the amino acid is histidine.

[0207] In some embodiments, the antioxidant is glutathione, ascorbic acid, cysteine, N-acetyl-L-tryptophanate, tocopherol, histidine, methionine, or a combination thereof. In certain embodiments, the antioxidant is methionine. In other embodiments, the protein is protamine, protamine sulfate, gelatin, or a combination thereof. In certain embodiments, the organic solvent is dimethyl sulfoxide, N-methyl-2-pyrrolidone, or a combination thereof. The paraben can be a parahydroxybenzoate. In still other embodiments, the bactericide is benzalkonium chloride (cationic surfactants), hypochlorites, peroxides, alcohols, phenolic compounds (e.g. carbolic acid), benzyl benzoate, or a combination thereof. In embodiments, the bactericide is benzyl benzoate.

[0208] In other embodiments, the fungicide is acibenzolar, 2-phenylphenol, anilazine, carvone, natamycin, potassium azide, or a combination thereof. In embodiments, the fungicide is benzyl benzoate. In certain embodiments, the vitamin is thiamine, riboflavin, niacin, pantothenic acid, biotin, vitamin Be, vitamin B12, folate, niacin, ascorbic acid, calciferols, retinols, quinones, or a combination thereof. In still other embodiments, the preservative is 704248642. vl5726.1016001sodium nitrate, sulfur dioxide, potassium sorbate, sodium sorbate, sodium benzoate, benzoic acid, methyl hydroxybenzoate, thimerosal, parabens, formaldehyde, castor oil, or a combination thereof. In embodiments, the preservative is methyl hydroxybenzoate, thimerosal, a paraben, formaldehyde, castor oil, or a combination thereof.

[0209] A number of nutrient media, preferably serum free, alone or in combination, may be used in the present disclosure, including commercially available media or other media well known in the art. Examples of such media (all without serum or having had the serum removed) include ADC-1, LPM (Bovine Serum Albumin-free), F10 (HAM), F12 (HAM), DCCM1, DCCM2, RPMI 1640, BGJ Medium (Fitton- Jackson Modification), Basal Medium Eagle (BME-with the addition of Earle's salt base), Dulbecco's Modified Eagle Medium (DMEM-without serum), Glasgow Modification Eagle Medium (GMEM), Leibovitz L-15 Medium, McCoy's 5 A Medium, Medium M199 (M199E-with Earle's salt base), Medium M199 (M199H- with Hank's salt base), Minimum Essential Medium Eagle (MEM-E- with Earle's salt base), Minimum Essential Medium Eagle (MEM-H- with Hank's salt base) and Minimum Essential Medium Eagle (MEM-NAA- with non-essential amino acids), among numerous others. In addition, serum-containing nutrient media may also be used in compositions according to the present disclosure, but the use of serum-containing media is less preferred because of the possibility that the serum may be contaminated with microbial agents and because the patient may develop immunological reactions to certain antigenic components contained in the serum.

[0210] In some embodiments, the oligopeptide is trileucine. In other embodiments, the biologic excipient are nucleic acids, oligonucleotides, antibodies or fragment thereof, amino acids, polyamino acids, peptides, proteins, cells, bacteria, gene therapeutics, genome engineering therapeutics, epigenome engineering therapeutics, hormones, nucleoproteins, glycoproteins, lipoproteins, exosomes, outer membrane vesicles, vaccines, viruses, bacteriophages, organelles, nutrient media, or a combination thereof. In certain embodiments, the chemical excipient are chemical drugs, contrast agents, dyes, magnetic particles, polymer beads, metal nanoparticles, metal microparticles, quantum dots, antioxidants, antibiotic agents, steroids, analgesics, local anesthetics, anti-inflammatory agents, parabens, anti-microbial agents, chemotherapeutic agents, vitamins, minerals, bactericides, antiseptics, or a combination thereof.

[0211] In other embodiments, the surfactant is polysorbate, magnesium stearate, sodium dodecyl sulfate, TRITON™ N-101, glycerin, poly oxy ethylated castor oil, docusate, sodium stearate, decyl glucoside, nonoxynol-9, cetyltrimethylammonium bromide, sodium bis(2- 714248642. vl5726.1016001ethylhexyl) sulfosuccinate, sodium laureth sulfate, lecithin, or a combination thereof. In some embodiments, the surfactant includes, but is not limited to: (i) cationic surfactants such as; cetyltrimethylammonium chloride, hexadecyltrimethylammonium chloride, benzalkonium chloride, benzethonium chloride, dioctadecyldimethylammonium bromide; (ii) anionic surfactants such as magnesium stearate, sodium dodecyl sulfate, dioctyl sodium sulfosuccinate, sodium myreth sulfate, perfluorooctanesulfonate, alkyl ether phosphates; (iii) non-ionic surfactants such as alkylphenol ethoxylates (TritonX-100), fatty alcohol ethoxylates (octaethylene glycol monododecyl ether, cocamide diethanolamine, poloxamers, glycerolmonostearate, fatty acid esters of sorbitol (sorbitan monolaurate, Tween 80, Tween 20; and (iv) zwitterionic surfactants such as cocamidopropyl hydroxysultaine, and 3-[(3-Cholamidopropyl)dimethylammonio]-1 -propanesulfonate (CHAPS). In other embodiments, the surfactant is polysorbate, magnesium stearate, sodium dodecyl sulfate, TRITON™ N-101, glycerin, polyoxyethylated castor oil, docusate, sodium stearate, decyl glucoside, nonoxynol-9, cetyltrimethylammonium bromide, sodium bis(2-ethylhexyl) sulfosuccinate, lecithin, sorbitan ester, phosphatidylcholine, polyglycerol polyricinoleate, siloxanes, cetyl polyethylene glycol / polypropylene glycol-10 / 1 dimethicone triglyceride, bis- polyethylene glycol / polypropylene glycol-14 / 14 dimethicone, bis-(glyceryl / lauryl) glyceryl lauryl dimethicone and caprylic / capric triglyceride, cetyl polyethylene glycol / polypropylene glycol-10 / 1 dimethicone, phospholipids, or a combination thereof. In certain embodiments, the surfactant is polysorbate, docusate, lecithin, sorbitan ester, or a combination thereof. In certain other embodiments, the surfactant is polysorbate 20, polysorbate 60, or polysorbate 80, e.g., Tween 20, Tween 60, Tween 80. In still other embodiments, the surfactant is polysorbate 20 or polysorbate 80. In certain embodiments, the fatty acid ester of sorbitol is a sorbitan ester, e.g., span 20, span 40, span 60, or span 80. In other embodiments, the surfactant is an ionic surfactant. In embodiments, the surfactant is polysorbate 80.

[0212] In some embodiments, a portion of the particles in a population have greater than about 50% therapeutic agent by weight (e.g. , after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent), e.g, greater than about 50, 55, 60, 65, 70, 75, 80, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 99.1, 99.2, 99.3, 99.4, 99.5, 99.6, 99.7, 99.8, 99.9% therapeutic agent by weight (e.g, after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In certain other embodiments, a portion of particles in the population have greater than about 80% therapeutic agent by weight (e.g., after removing an aqueous fraction from a droplet, after dehydrating 724248642. vl5726.1016001droplets within a liquid stream to form particles comprising a therapeutic agent). In other embodiments, a portion of particles in the population have greater than about 90% therapeutic agent by weight (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In certain embodiments, a portion of particles in the population have greater than about 95% therapeutic agent by weight (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In still other embodiments, a portion of particles in the population have greater than about 98% therapeutic agent by weight (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In certain embodiments, a portion of particles in the population have greater than about 99% therapeutic agent by weight (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent).

[0213] In certain embodiments, a portion of particles in a population include a loading of therapeutic agent from at least about 1 to about 100 wt% (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent), e.g., from at least about 50 to about 100 wt%, from at least about 75 to about 100 wt%, from at least about 90 to about 100 wt%, from at least about 95 to about 100 wt%, from at least about 99 to about 100 wt%, or from at least about 99.9 to about 100 wt% e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). At these loadings the therapeutic agent retains from at least about 0.5 to about 1.0 activity during particle formation, e.g., from at least about 0.75 to about 1.0 activity, from at least about 0.9 to about 1.0 activity, from at least about 0.95 to about 1.0 activity, from at least about 0.99 to about 1.0 activity, or from at least about 0.999 to about 1.0 activity. This includes the activity retained through primary desiccation and, in some cases, secondary desiccation post processing.

[0214] In other embodiments, a portion of particles in a population have less than 25% increased aggregation or less than 25% increased fragmentation of the therapeutic agent compared with an aqueous feed solution before forming the particles (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent), e.g., less than about 25, 20, 15, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1.9, 1.8, 1.7, 1.6, 1.5, 1.4, 1.3, 1.2, 1.1, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, or 0.1% of the therapeutic agent compared with an aqueous feed solution before forming the particles (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid 734248642. vl5726.1016001stream to form particles comprising a therapeutic agent). In certain embodiments, a portion of particles in a population have about 10% to about 1% increased aggregation of the therapeutic agent compared with the aqueous feed solution before forming the portion of the particles (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In certain other embodiments, a portion of particles in a population have about 1% to about 0.5% increased aggregation of the therapeutic agent compared with the aqueous feed solution before forming the particles (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In embodiments, a portion of particles in a population are substantially free from any additional aggregation of the therapeutic agent compared with an amount of aggregation present in the aqueous feed solution before forming the particles (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In still other embodiments, a portion of particles in a population have less than about 25%, about 20%, about 15%, about 10%, about 5%, about 3%, about 2%, about 1%, or about 0.5% increased fragmentation of the therapeutic agent compared with the aqueous feed solution before forming the particles (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In certain embodiments, a portion of particles in a population are substantially free from any fragmentation of the therapeutic agent compared with the aqueous feed solution before forming the particles (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). The process of forming the particles may be gentle and / or reduce total aggregation of the therapeutic compared to the amount of aggregation present in the aqueous feed solution. In some embodiments, the amount of aggregation present in the suspension of particles may be at least about 0.5%, at least about 1%, at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, or at least about 50% reduced fragmentation of the therapeutic agent compared with the aqueous feed solution before forming the particles. Suitable methods for measuring aggregation and fragmentation of a biologic can be accomplished by using size-exclusion chromatography (SEC).

[0215] In some embodiments, the process of particle formation provides less than a 50% change in charge variants of a therapeutic agent in at least a portion of a population of particles (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a 744248642. vl5726.1016001liquid stream to form particles comprising a therapeutic agent), e.g., an antibody, (e.g., less than 40, 30, 25, 20, 15, 10, 8, 5, 4, 3, 1, or 0.5% (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent)) as compared to the therapeutic agent prior to particle formation. Charge variants may be acidic, basic, or neutral, and the variation may be caused post-translation modifications at terminal amino acids, such as asparagine deamidation or lysine glycation. For example, charge variants include the loss of a positive charge by the loss of a C-terminal lysine residue, covalent bonding of the amine portions of two lysine residues by reducing sugars, or the conversion of an N-terminal amine to a neutral amide by the cyclization of N-terminal glutamines. Negative charges on proteins, e.g., antibodies, can appear by the conversion of asparagine residues to aspartic acid and / or isoaspartic residues via a deamidation reaction. Example methods of measuring charge variants include cation exchange chromatography (CIEX), where the variants are quantified by dividing the area under the peak corresponding to the variant, e.g., acidic, basic, or neutral population by the cumulative area contained beneath all peaks in the sample spectrum. Changes in charge variant population percentage between two samples, e.g., Sample A and Sample B, are computed as the numerical difference in the respective population variant percentages, i.e., by subtracting the specific variant, e.g., acidic, percentage of Sample B from the specific variant, e.g., acidic, percentage of Sample A, or vice versa. In certain embodiments, the analysis may be extended similarly for all variants within a population.

[0216] In certain embodiments, a portion of particles in a population have less than about 50% change in charge variants of the therapeutic agent (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent), e.g., less than about 45, 40, 35, 30, 25, 20, 15, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, 0.5, 0.1% (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent), compared to the starting biologic prior to particle formation. In embodiments, a portion of the particles in the population are substantially free from any change in charge variants of the therapeutic agent (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent), compared to the starting biologic prior to particle formation. Suitable methods for measuring a change in charge variants of a biologic can be accomplished by using cation exchange chromatography (CIEX).

[0217] In some embodiments, a portion of particles in a population have a surfactant content of less than about 10% by mass (e.g., after removing an aqueous fraction from a droplet,754248642. vl5726.1016001after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent), e.g., less than about 9, 8, 7, 6, 5, 4, 3, 2, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.09, 0.08, 0.07, 0.06, 0.05, 0.04, 0.03, 0.02, 0.01, 0.009, 0.008, 0.007, 0.006, 0.005, 0.004, 0.003, 0.002, 0.001% by mass (e.g, after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In other embodiments, a portion of the particles in a population have a surfactant content of less than about 5% by mass (e.g, after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In certain embodiments, a portion of the particles in a population have a surfactant content of less than about 3% by mass (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In still other embodiments, a portion of the particles in a population have a surfactant content of less than about 1% by mass (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In certain other embodiments, a portion of the particles in a population have a surfactant content of less than about 0.1% by mass (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In some embodiments, a portion of the particles in a population have a surfactant content of less than about 0.01% by mass (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In other embodiments, a portion of the particles in a population have a surfactant content of less than about 0.001% by mass (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In embodiments, a portion of the particles in a population are substantially free from any surfactant content (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent).

[0218] In some embodiments, a portion of particles in a population have a surfactant content of at least about 10% by mass (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent), e.g., at least about 9, 8, 7, 6, 5, 4, 3, 2, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.09, 0.08, 0.07, 0.06, 0.05, 0.04, 0.03, 0.02, 0.01, 0.009, 0.008, 0.007, 0.006, 0.005, 0.004, 0.003, 0.002, 0.001% by mass (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In other embodiments, a portion of the particles in a population have a surfactant content of at 764248642. vl5726.1016001least about 5% by mass (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In certain embodiments, a portion of the particles in a population have a surfactant content of at least about 3% by mass (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In still other embodiments, a portion of the particles in a population have a surfactant content of at least about 1% by mass (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In certain other embodiments, a portion of the particles in a population have a surfactant content of at least about 0.1% by mass (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In some embodiments, a portion of the particles in a population have a surfactant content of at least about 0.01% by mass (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In other embodiments, a portion of the particles in a population have a surfactant content of at least about 0.001% by mass (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent).

[0219] In other embodiments, the surfactant content of a portion of particles in a population is from 0 to 10 wt% (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent), e.g., from 0 to 5 wt%, from 0 to 3 wt%, from 0 to 2 wt%, from 0 to 1 wt%, from 0 to 0.5 wt%, from 0 to 0.2 wt%, from 0 to 0.1 wt%, from 0 to 0.01 wt%, or from 0 to 0.001 wt% (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In certain embodiments, the surfactant content of a portion of the particles in a population is at least about 0 wt% (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent), e.g., at least about 5 wt%, at least about 3 wt%, at least about 2 wt%, at least about 1 wt%, at least about 0.5 wt%, at least about 0.2 wt%, at least about 0.1 wt%, at least about 0.01 wt%, or at least about 0.001 wt% (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). Example methods of measuring the surfactant content include reconstitution of a portion of the particles in a population in an appropriate medium, e.g., deionized water, and subsequent analysis of the reconstituted solution through774248642. vl5726.1016001liquid chromatography. The chromatographic technique may include the use of a charged aerosol detector (CAD) or an evaporative light scattering detector (ELSD).

[0220] In some embodiments, the aqueous liquid content remaining in a portion of particles in a population are less than about 25% by mass, e.g., less than about 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1.9, 1.8, 1.7, 1.6, 1.5, 1.4, 1.3, 1.2, 1.1, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, or 0.1% by mass remaining (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In other embodiments, a portion of the particles in a population have less than about 15% of residual aqueous fraction by mass remaining (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In still other embodiments, a portion of the particles in a population have less than about 5% of residual aqueous fraction by mass remaining (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In certain embodiments, a portion of the particles in a population have less than about 1% of residual aqueous fraction by mass remaining (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In embodiments, a portion of the particles in a population are substantially free from any residual aqueous fraction by mass (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent).

[0221] In other embodiments, a portion of particles in a population have about 1% to about 15% of residual aqueous fraction by mass remaining (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In still other embodiments, a portion of the particles in a population have about 1% to about 10% of residual aqueous fraction by mass remaining (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In certain embodiments, a portion of the particles in a population have about 1% to about 5% of residual aqueous fraction by mass remaining (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In certain embodiments, a portion of the particles in a population have about 1% to about 3% of residual aqueous fraction by mass remaining (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In embodiments, a 784248642. vl5726.1016001portion of the particles in a population are substantially free from any residual aqueous fraction by mass (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). Example methods for the measurement of residual aqueous fraction and / or organic liquid content include chemical titration methods, e.g., Karl Fischer titration involving an oven. A variety of solvents, including water, may also be measured using weight loss methods involving thermal excitation. Example methods include Thermogravimetric Analysis with Infrared Spectroscopy (TGA-IR) or Gas Chromatography Flame Ionization Detector Mass Spectrometry (GC-FID / MS).

[0222] As used herein, the terms “moisture content” and “water content” are used interchangeably. In some embodiments, the volume-to-weight percent average moisture e.g. water content of a portion of particles in a population is less than about 30% by mass (e.g, after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent), e.g, less than about 30, 25, 20, 15, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1.9, 1.8, 1.7, 1.6, 1.5, 1.4, 1.3, 1.2, 1.1, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, or 0.1% by mass (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In other embodiments, a portion of the particles in a population have a volume-weighted average of less than about 15% residual moisture by mass (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In certain embodiments, a portion of the particles in a population have a volume-weighted average of less than about 10% residual moisture by mass (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In other embodiments, a portion of the particles in a population have a volume-weighted average of less than about 5% residual moisture by mass (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In still other embodiments, a portion of the particles in a population have a volume-weighted average of less than about 3% residual moisture by mass (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In certain embodiments, a portion of the particles in a population have a volume-weighted average of less than about 2% residual moisture by mass (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In embodiments, a portion of the particles in a population have a volume-weighted average of less than about 1% residual moisture by mass 794248642. vl5726.1016001(e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In certain embodiments, a portion of the particles in a population are substantially free from any residual moisture by mass (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In some embodiments, a portion of the particles in a population have a volume-weighted average of less than about 0.1% of residual moisture by mass e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In some embodiments, a portion of the particles in a population have a volume-weighted average of less than about 0.01% of residual moisture by mass e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). Example methods for the measurement of moisture content include chemical titration methods, e.g., Karl Fischer titration involving an oven. A variety of solvents, including water, may also be measured using weight loss methods involving thermal excitation. Example methods include Thermogravimetric Analysis with Infrared Spectroscopy (TGA-IR) or Gas Chromatography Flame Ionization Detector Mass Spectrometry (GC-FID / MS).

[0223] In some embodiments, the volume-weighted average organic liquid content remaining in a portion of particles in a population is less than about 30% by mass, e.g., less than about 30, 25, 20, 15, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1.9, 1.8, 1.7, 1.6, 1.5, 1.4, 1.3, 1.2, 1.1, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, or 0.1% by mass remaining (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In other embodiments a portion of the particles in a population have a volume-weighted average of less than about 15% of residual organic liquid by mass remaining (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In certain embodiments, a portion of the particles in a population have a volume-weighted average of less than about 10% of residual organic liquid by mass remaining (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In other embodiments, a portion of the particles in a population have a volume-weighted average of less than about 5% of residual organic liquid by mass remaining (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In still other embodiments, a portion of the particles in a population have a volume-weighted average of less 804248642. vl5726.1016001than about 3% of residual organic liquid by mass remaining (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In certain embodiments, a portion of the particles in a population have a volume-weighted average of less than about 1% of residual organic liquid by mass remaining (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In embodiments, a portion of the particles in a population are substantially free from any residual organic liquid by mass e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent).

[0224] In other embodiments, a portion of particles in a population has a volume-weighted average of about 1% to about 15% residual moisture by mass e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In still some embodiments, a portion of the particles in a population have a weighted volume average of about 1% to about 10% residual moisture by mass e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In some embodiments, a portion of the particles in a population have a weighted volume average of about 1% to about 5% residual moisture by mass e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In certain embodiments, a portion of the particles in a population have a weighted volume average of about 1% to about 3% residual moisture by mass e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In embodiments, a portion of the particles in a population are substantially free from any residual moisture by mass e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In other embodiments, a portion of the particles in a population are stable for at least one month. In some embodiments, a portion of the particles in a population are stable for at least two months. In certain embodiments, a portion of the particles in a population are stable for at least three months. In certain other embodiments, a portion of the particles in a population are stable for at least three months at 40 °C. In certain other embodiments, a portion of the particles in a population are stable for at least six months at 4 °C.

[0225] As used herein, the term “dispersity index” (DI) is a parameter characterizing the degree of non-uniformity of a size distribution of particles. The poly dispersity index (PDI), “population dispersity” or “span”, e.g., D10, D50, D90, is a value that can indicate the breadth 814248642. vl5726.1016001of the particle size distribution. Particle size distribution are reported by DIO, D50, D90, and the mean particle size in pm, with the values representing the percentage of particles that are smaller than the indicated D-number, e.g. the DIO particle size is the particle diameter at which 10% of the mass is composed of particles with a diameter less than this value, the D50 particle size is the particle diameter at which 50% of the mass is composed of particles with a diameter less than this value and the D90 particle size is the particle diameter at which 90% of the mass is composed of particles with a diameter less than this value. The DIO, D50, and D90 particle size distribution can be measured using a laser light scattering particle sizer.

[0226] In other embodiments described herein, a portion of particles in a population have a poly dispersity index from about 0.002 to about 1.000, e.g., from about 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, 0.010, 0.020, 0.030, 0.040, 0.050, 0.060, 0.070, 0.080, 0.090, 0.100, 0.200, 0.300, 0.400, 0.500, 0.600, 0.700, 0.800, 0.900 to about 1.000. In certain embodiments, a portion of the particles in a population have a poly dispersity index from about 0.002 to about 0.900. In certain embodiments, a portion of the particles in a population have a poly dispersity index from about 0.100 to about 0.300.

[0227] In certain embodiments, a particle may comprise one or more therapeutic agents (e.g., two therapeutic agents). In some embodiments, a particle comprises two or more therapeutic agents. In certain embodiments, a particle comprises two or more active pharmaceutical ingredients comprising at least one biologic and at least one small molecule. In some embodiments, a plurality of particles is provided, wherein each particle within the plurality of particles comprises the same one or more therapeutic agents (e.g., two biologic agents). In some embodiments, a plurality of particles is provided, wherein at least two particles within the plurality of particles comprise one or more different therapeutic agents. In certain embodiments, at least one therapeutic agent in a particle or plurality of particles is a hyaluronan degrading agent, such as hyaluronidase. In some embodiments, the hyaluronidase is rHuPH20 or is a modified PH20 hyaluronidase, such as those set forth in PCT / US2012 / 072182 and PCT / US2024 / 045720, the contents of each of which are hereby incorporated by reference in their entireties.

[0228] In some embodiments, a plurality of particles can have a volume-weighted average diameter of at least about 5 pm, e.g., at least about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75 pm. In some embodiments, a plurality of the particles can have a volume-weighted average diameter of at least about 20 pm. In some embodiments, a plurality 824248642. vl5726.1016001of the particles can have a volume-weighted average diameter of at least about 15 pm. In some embodiments, a plurality of the particles can have a volume-weighted average diameter of at least about 10 pm. In some embodiments, a plurality of the particles can have a volume-weighted average diameter of at least about 5 pm.

[0229] In some embodiments, a portion of particles in a population can have a volume-weighted average diameter of at least about 5 pm, e.g., at least about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75 pm. In some embodiments, a portion of the particles in a population can have a volume-weighted average diameter of at least about 20 pm. In some embodiments, a portion of the particles in a population can have a volume-weighted average diameter of at least about 15 pm. In some embodiments, a portion of the particles in a population can have a volume-weighted average diameter of at least about 10 pm. In some embodiments, a portion of the particles in a population can have a volume-weighted average diameter of at least about 5 pm.

[0230] In other embodiments, a portion of particles in a population can have a volume-weighted average diameter from at least about 0.1 to about 1000 pm (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent), e.g, at least about 0.1 to about 75 pm, at least about 1 to about 100 pm, at least about 100 to about 250 pm, or at least about 0.1 to about 1 pm (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). In still other embodiments, a portion of the particles in a population can have a size dispersity from at least about 0 to about 0.9 e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent), e.g, from at least about 0 to about 0.7, from at least about 0 to about 0.5, or from at least about 0 to about 0.2 (e.g., after removing an aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent). Example methods of measuring the particle size and distribution include imaging flow cytometry, laser diffraction, and image analysis of scanning electron micrographs of the particles in which an average spherical radius or diameter can be calculated on the basis of the cross-sectional areas of the particles projected onto the plane of the image. In certain other embodiments of the present disclosure, a portion of particles in a population can have a volume-weighted average diameter of at least about 0.1 to about 1000 pm, and a glass transition temperature of about 0 to about 250 °C, (e.g., after removing an 834248642. vl5726.1016001aqueous fraction from a droplet, after dehydrating droplets within a liquid stream to form particles comprising a therapeutic agent).

[0231] The particles comprising at least one therapeutic agent described herein, can be prepared in a number of ways, as well as any methods of forming the particles disclosed in, for example, in International Application Nos. PCT / US2017 / 063150 (Pub. No. WO 2018 / 098376), PCT / US2018 / 043774 (Pub. No. WO 2019 / 023392), PCT / US2019 / 033875 (Pub. No. WO 2019 / 226969), PCT / US2020 / 15957 (Pub. No. WO 2020 / 160323), PCT / US2020 / 050508 (Pub. No. WO 2021 / 050953), PCT / US2021 / 16878, and PCT / US2021 / 018806, the contents of each of which are hereby incorporated by reference in their entireties.Pharmaceutical Compositions

[0232] Injectable particle suspensions can exhibit variations in sedimentation, which may impact the use or administration of the drug composition. In particular, high concentration, low volume and low syringe force injectable particle suspensions of therapeutic biologies will settle or sediment out of the suspension medium over some period of time, thus, requiring premixing or resuspension prior to injection. The sedimentation of high particle concentrations at low delivery volumes may also lead to high injection forces and propagate decomposition of the therapeutic biologic in the composition. As used herein, the term “sediment” or “sedimentation” is the process of particles settling or being densely deposited at the bottom of a container closure, e.g., vial, cartridge, syringe, portable drug delivery injection device, or the like. The settling of densely deposited particles generally leads to excessively high injection forces or syringe blockage and requires manual agitation or premixing prior to administration of the composition. In some embodiments, a container is a vial, cartridge, or syringe.

[0233] Flocculation or “particle flocculation” is a phenomenon that arises from the interplay between the interfacial chemistry and environmental conditions that govern particleparticle interactions. Flocculation volume, as used herein, is a measure of the amount of particle agglomerates that are occupied by the particulate dispersion, expressed as a percentage of the total fluid volume. More specifically, the flocculation volume (F) is the ratio of the volume of particle agglomerates (Vu) to the total fluid volume (Vo). The flocculation volume can decrease over time based on the sedimentation properties of the particle agglomerates. In a flocculated suspension, the particles undergo collisions leading to particle agglomeration or agglomerates, that increase in size, e.g., increase in flocculation volume. The term “agglomerates” refers to a cluster of particles that are loosely coherent. The loose networks of particle agglomerates maintain enough particle-to-particle distance to prevent particle844248642. vl5726.1016001sedimentation that can lead to high injection forces. When the flocculation volume is equal to 1, the yield stress (the stress at which the flocculation volume will undergo deformation) can be measured to indicate the shear stress that is needed to make the suspension flow. Measurements for yield stress can be acquired by a parallel plate rheometer (ANTON PAAR™ MCR 92). An increase in yield stress indicates an increase in agglomerates. Other approaches for measuring agglomerates include, but are not limited to, laser diffraction, relaxation NMR, microscopy and small-angle scattering.

[0234] In compositions disclosed herein, the process of agglomeration may be controlled by the use of flocculation agents. Flocculation agents, or flocculating agents, are chemicals that promote flocculation by causing suspended particles in the liquid to agglomerate, thereby forming agglomerates. In non-aqueous particle suspensions, the breakup and redispersion of agglomerates is governed by the shear stress imparted on the flocculated particle system. Shear stress is caused by a force acting on the material’s surface, which causes deformation. In these systems, shear can be attributed to the movement of particles or agglomerates through a velocity gradient in the non-aqueous suspending medium or near any interface (e.g., the container closure, other particles, etc.). Shear stress can be introduced to the flocculation volume, for example, mechanically, and can be controlled experimentally.

[0235] Compositions disclosed herein may comprise a flocculating agent and are generally high concentration, low volume and low syringe force injectable particle suspensions of therapeutic biologies that permit administration without the need for manual agitation or premixing prior to injection or administration of the composition. The compositions beneficially maintain stable flocculation volumes that avoid particle sedimentation that can result in blockage of syringes, portable drug delivery injection devices, or orally dosed liquid injector capsules.

[0236] In some embodiments, pharmaceutical compositions for administration include non-aqueous solutions of the active therapeutic biologies in water-soluble form. Preferably, suspensions of the active therapeutic biologies may be prepared as appropriate oily injection compositions. Suitable lipophilic solvents or vehicles include fatty oils (e.g., sesame oil, corn oil), or fatty acid esters (e.g., ethyl oleate or triglycerides), or liposomes. The proper viscosity can be maintained by the maintenance of the required particle size in the case of injection, and by the use of a flocculation agents. The prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like.854248642. vl5726.1016001

[0237] In a composition as described herein, the process of agglomeration can be controlled by the use of flocculation agents which can permit administration of particle compositions without the need for manual agitation or premixing prior to injection or administration. Flocculation agents suitable for use in the methods described herein are described in PCT / US2023 / 066395, which is incorporated by reference herein in its entirety. Examples of flocculation agents include, but are not limited to, polysorbate 20, polysorbate 60, polysorbate 80, PEG 300, PEG 400, PEG 600, ethylene glycol, propylene glycol, or a combination thereof. In some embodiments, the addition of a flocculation agent to the particle suspension does not substantially increase viscosity of the composition. As disclosed herein, the addition of a flocculation agent to a suspension of particles increases the flocculation volume of the suspension of the particles. In certain embodiments, the addition of a flocculation agent to a suspension of particles increases the stability of the flocculation volume for at least one month. In some embodiments, the addition of a flocculation agent to a suspension of particles reduces the syringe force. In some embodiments, the addition of a flocculation agent to a suspension of particles improves the pharmacokinetics (PK) of administration (e.g., subcutaneous administration) of microparticle (e.g., Ab microparticle) suspensions and shows higher bioavailability than the administration e.g., subcutaneous administration) of aqueous Ab solution. In some embodiments, the addition of a flocculation agent to a suspension of particles increases the stability of the proteins in the particles.

[0238] In some embodiments, the composition described herein, use a concentration of the flocculation agent in the suspension of at least about 50 mg / mL, e.g., at least about 45, 40, 35, 30, 25, 20, 15, 10, 5, 3, 1, 0.5, 0.1, 0.05, or 0.01 mg / mL. In some embodiments, the concentration of the flocculation agent in the suspension is at least about 10 mg / mL. In some embodiments, the concentration of the flocculation agent in the suspension is at least about 5 mg / mL. In some embodiments, the concentration of the flocculation agent in the suspension is at least about 3 mg / mL. In particular embodiments, the concentration of the flocculation agent in the suspension is at least about 1 mg / mL. In some embodiments, the concentration of the flocculation agent in the suspension is at least about 0.1 mg / mL. In certain embodiments, the concentration of the flocculation agent in the suspension is at least about 0.01 mg / mL.

[0239] Viscosity can play an important role in the handling and administration of inj ectable products. For suspension products, high viscosities drug products may be difficult to deliver through a needle (e.g., 27-gauge needle) since it takes greater force to actuate the injection device, e.g., syringe, portable drug delivery injection device, or orally dosed liquid injector864248642. vl5726.1016001capsules. Alternatively, using a larger gauge needle or requiring longer injection times can reduce patient compliance to therapy.

[0240] In some embodiments, the composition has a viscosity (e.g., an apparent viscosity) of less than about 500 mPa s, less than about 400 mPa s, less than about 300 mPa s, less than about 200 mPa s, less than about 175 mPa s, less than about 150 mPa s, less than about 125 mPa s, less than about 100 mPa s, less than about 95 mPa s, less than about 90 mPa s, less than about 85 mPa s, less than about 80 mPa s, less than about 75 mPa s, less than about 70 mPa s, less than about 65 mPa s, less than about 60 mPa s, less than about 55 mPa s, less than about 50 mPa s, less than about 45 mPa s, less than about 40 mPa s, less than about 35 mPa s, less than about 30 mPa s, less than about 25 mPa s, less than about 20 mPa s, less than about 19 mPa s, less than about 18 mPa s, less than about 17 mPa s, less than about 16 mPa s, less than about 15 mPa s, less than about 14 mPa s, less than about 13 mPa s, less than about 12 mPa s, less than about 11 mPa s, less than about 10 mPa s, less than about 9.5 mPa s, less than about 9 mPa s, less than about 8.5 mPa s, less than about 8 mPa s, less than about 7.5 mPa s, less than about 7 mPa- s, less than about 6.5 mPa- s, less than about 6 mPa- s, less than about 5.5 mPa s, less than about 5 mPa s, or less than about 1 mPa s (one millipascal-second). In certain embodiments, the composition’s viscosity (e.g., apparent viscosity) may be measured at a syringe needle. The apparent viscosity is the shear stress applied to a fluid divided by the shear rate. Mixtures of liquids may also be used to control viscosity. The units “mPa s” and “cP” are used herein interchangeably in the broadest sense.

[0241] In some embodiments, the composition has an apparent viscosity of less than about 500 mPa s (e.g., less than about 500, about 400, about 300, about 200, about 150, about 100, about 75, about 50, about 25, about 20, about 15, about 10, about 5, about 1 mPa s). In some embodiments, the composition has an apparent viscosity of less than about 80 mPa- s. In certain embodiments, the composition has an apparent viscosity of less than about 50 mPa s. In some embodiments, the composition has an apparent viscosity of less than about 40 mPa- s. In certain embodiments, the composition has an apparent viscosity of less than about 30 mPa s. In particular embodiments, the composition has an apparent viscosity of less than about 25 mPa- s. In some embodiments, the composition has an apparent viscosity of less than about 20 mPa s. In certain embodiments, the composition has an apparent viscosity of less than about 15 mPa s. In some embodiments, the composition has an apparent viscosity of less than about 10 mPa s. In certain embodiments, the composition has an apparent viscosity of less than about 5 mPa s.

[0242] In some embodiments, the composition’s viscosity (e.g., apparent viscosity) is greater than about 500 mPa s (e.g., greater than about 500 mPa s, about 1,000 mPa s, about 874248642. vl5726.10160012,500 mPa s, about 5,000 mPa s, about 7,500 mPa s, about 10,000 mPa s). In some embodiments, the composition has an apparent viscosity of greater than about 10,000 mPa s. In some embodiments, the composition has an apparent viscosity of greater than about 7,500 mPa s. In some embodiments, the composition has an apparent viscosity of greater than about 5,000 mPa s. In some embodiments, the composition has an apparent viscosity of greater than about 2,500 mPa - s. In some embodiments, the composition has an apparent viscosity of greater than about 1,000 mPa s. In some embodiments, the composition has an apparent viscosity of greater than about 500 mPa s.

[0243] In some embodiments, the composition may be injected using less than about 1000 N of syringe force (e.g., less than about 1000 N, about 900 N, about 800 N, about 700 N, about 600 N, about 500 N, about 400 N, about 300 N, about 250 N, about 200 N, about 150 N, about 125 N, about 115 N, about 110 N, about 105 N, about 100 N, about 95 N, about 90 N, about 85 N, about 80 N, about 75 N, about 70 N, about 65 N, about 60 N, about 55 N, about 50 N, about 45 N, about 40 N, about 35 N, about 30 N, about 25 N, about 20 N, about 15 N, about 10N, about 9 N, about 8 N, about 7 N, about 6 N, about 5 N, about 4 N, about 3 N, about 2 N, about 1 N). In certain embodiments, the composition may be injected in less than about 20 seconds (e.g., less than about 20, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1 seconds). In some embodiments, the composition may be injected in less than about 7 seconds.

[0244] As described herein, a higher flocculation volume indicates increased agglomerates or less particle sedimentation, and accordingly, improved injection force performance due to more uniform dispersion of the particles in a liquid carrier e.g., propylene glycol diesters, ethyl oleate). In some embodiments, the composition has a flocculation volume greater than about 50% after initial mixing, e.g., 60%, 70%, 75%, 80%, 85%, 90%, 95% or greater than about 98% after initial mixing. In some embodiments, the composition has a flocculation volume greater than about 70% after initial mixing. In certain embodiments, the composition has a flocculation volume greater than about 80% after initial mixing. In particular embodiments, the composition has a flocculation volume greater than about 85% after initial mixing. In certain embodiments, the composition has a flocculation volume of about 100% after initial mixing. Initial mixing refers to the homogenization of an initially heterogeneous particle suspension with a flocculating agent through bulk motion creating a flocculation volume.

[0245] In some embodiments, the flocculation volume is reduced by less than about 20%, e.g., 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1%, after at least one month under container closure storage conditions at less than about 40 °C. In some embodiments, the flocculation volume is reduced by less than about 7% after at least one month under container 884248642. vl5726.1016001closure storage conditions at less than about 40 °C. In certain embodiments, the flocculation volume is reduced by less than about 5% after at least one month under container closure storage conditions at less than about 40 °C. In some embodiments, the flocculation volume is reduced by less than about 3% after at least one month under container closure storage conditions at less than about 40 °C. In certain embodiments, the flocculation volume is reduced by less than about 1% after at least one month under container closure storage conditions at less than about 40 °C. In some embodiments, the plurality of particles and flocculation agent remain substantially suspended in the liquid carrier for at least one month. In some embodiments, the composition has substantially the same flocculation volume for at least one month. In some embodiments, the composition does not require resuspension.

[0246] In certain embodiments, the disclosure relates to a composition, e.g., pharmaceutical composition, comprising a plurality of particles suspended in a pharmaceutically acceptable liquid carrier with a flocculation agent (e.g., a flocculation agent disclosed herein), wherein the particles (e.g., substantially all of the particles) comprise at least one therapeutic biologic (e.g., a therapeutic biologic disclosed herein).

[0247] In compositions disclosed herein, the process of agglomeration can be controlled by altering the surface charge on a portion of the particles (e.g., microparticles) in a suspension. In some embodiments, the surface charge is adjusted by modifying the pH of the aqueous feed solution. An increase in surface charge can lead to particle repulsion and excluded volume effects. As a result, the material exhibits an appreciable yield stress, preventing the particles from settling under gravity.

[0248] In certain embodiments according to the disclosure as described herein, a composition comprising a plurality of particles can have improved stability of the therapeutic agent compared to an aqueous composition comprising the therapeutic agent in monomeric form.

[0249] In other embodiments, the disclosure provides a composition containing a plurality of particles that include a therapeutic agent, e.g., an antibody, antibody fragment, or human serum albumin (HSA), where the storage stability of the therapeutic agent in the particles is improved with respect to the storage stability of the therapeutic agent in the feed solution. In some embodiments, storage conditions are defined by time (e.g., more than about 2 years, more than about 1 year, more than about 6 months, more than about 3 months, more than about 1 month, or more than about 1 week) and temperature (e.g., about -80 °C to about 100 °C, about -80 °C to about 60 °C, about -20°C to about 60°C, about 4 to about 60 °C), among potentially other variables. In still other embodiments, the storage time is about 3 days, about 7 days, about 894248642. vl5726.101600130 days, about 90 days, about 180 days, about 1 year, or about 2 years. In certain embodiments, this temperature is about -80 °C, about -40 °C, about -20 °C, about 4 °C, about 25 °C, about 40 °C, or about 40 to about 60 °C.

[0250] The phrase “pharmaceutically acceptable” is employed herein, to refer to those therapeutic agents, materials, compositions, and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio. The term “pharmaceutically acceptable” can refer to particles and compositions comprising a plurality of particles that do not produce an adverse, allergic, or other untoward reaction when administered to a mammal, such as a human, as appropriate. The preparation of a pharmaceutical composition comprising an antibody or additional active ingredient will be known to those of skill in the art in light of the present disclosure. Moreover, for mammal (e.g., human) administration, it will be understood that preparations should meet sterility, pyrogenicity, general safety, and purity standards as required by the FDA Office of Biological Standards.

[0251] The phrase “pharmaceutically acceptable liquid” includes any and all aqueous solvents (e.g., water, alcoholic / aqueous solutions, saline solutions, parenteral vehicles, such as sodium chloride, Ringer's dextrose, and the like), non-aqueous solvents (e.g., propylene glycol, polyethylene glycol, vegetable oil, and injectable organic esters, such as ethyl oleate), dispersion media, coatings, surfactants, antioxidants, preservatives (e.g., antibacterial or antifungal agents, anti-oxidants, chelating agents, and inert gases), isotonic agents, absorption delaying agents, salts, drugs, drug stabilizers, gels, binders, excipients, disintegration agents, lubricants, sweetening agents, flavoring agents, dyes, fluid and nutrient replenishers, such like materials and combinations thereof, as would be known to one of ordinary skill in the art. The pH and exact concentration of the various components in a pharmaceutical composition are adjusted according to well-known parameters. In certain embodiments, the plurality of particles is suspended in a pharmaceutically acceptable liquid. In embodiments, the liquid is a pharmaceutically acceptable liquid.

[0252] A pharmaceutical composition (formulation) as described herein, can be administered to a subject by any of a number of routes of administration including, for example, parenterally (including intramuscularly, intravenously, subcutaneously or intrathecally as, for example, a sterile solution or suspension); intraperitoneally; or subcutaneously. In certain embodiments, a composition may be suspended in a non-aqueous carrier for subcutaneous delivery. Details of appropriate routes of administration and compositions suitable for same 904248642. vl5726.1016001can be found in, for example, U.S. Patent Nos. 6,110,973; 5,763,493; 5,731,000; 5,541,231; 5,427,798; 5,358,970 and 4,172,896, as well as in patents cited therein. The term “suspension formulation” refers to a liquid formulation including particles disposed within a carrier liquid in which they are not soluble on an appropriate timescale. The particles may settle over time, / .< ., the physical stability of the suspension is not indefinite, but may be re-suspended using a form of agitation or excitation.

[0253] A “therapeutic amount” refers to an amount of a therapeutic agent required to produce the desired effect. As used herein, the terms “treat,” “treated,” and “treating” mean both therapeutic treatment and prophylactic or preventative measures wherein the object is to prevent or slow down (lessen) an undesired physiological condition, disorder, or disease, or obtain beneficial or desired clinical results. Beneficial or desired clinical results include, but are not limited to, alleviation of symptoms; diminishment of the extent of a condition, disorder, or disease; stabilized ( / .< ., not worsening) state of condition, disorder, or disease; delay in onset or slowing of condition, disorder, or disease progression; amelioration of the condition, disorder, or disease state or remission (whether partial or total), whether detectable or undetectable; an amelioration of at least one measurable physical parameter, not necessarily discernible by the patient; or enhancement or improvement of condition, disorder, or disease. Treatment includes eliciting a clinically significant response without excessive levels of adverse events and other side effects. Treatment also includes prolonging survival as compared to expected survival if not receiving treatment.

[0254] In some embodiments, insoluble particulate matter with characteristic sizes greater than or equal to about 100 pm that persist upon dissolution in an aqueous feed solution are referred to as Visible Particles (VP). In embodiments of the present disclosure described herein, the composition is substantially free of Visible Particles (VP). In certain embodiments, the aqueous feed solution is water, aqueous buffer or a physiologically relevant aqueous feed solution. In other embodiments, insoluble particulate matter which is visible to the naked eye under prescribed lighting conditions persist upon reconstitution of the particles of the present disclosure into a liquid pharmaceutical composition. Insoluble particulates of this type, is sometimes referred to as Visible Particles (VPs), and are typically greater than about 100 pm in size. VPs are present in quantities from about 0 to about 1 per about 1 mL, e.g., from about 0 to about 0.01 per about 1 mL, from about 0 to about 0.001 per about 1 mL, or about 0 to about 0.0001 per about 1 mL. Example methods of measuring VPs include analysis of the therapeutic agent by visual inspection against and black and white background for 5 seconds under illumination of about 2000 and about 3750 lux in accordance with USP <790> after 914248642. vl5726.1016001reconstitution and dilution of the therapeutic agent to a standard concentration, e.g., about 100 mg / mL or about 1 mg / mL. In some embodiments, fewer than 65 samples in 10,000 (0.65%) are rejected on the basis of USP <790>. Alternate inspection strategies light-obscuration, automated optical imaging systems, or X-ray imaging in accordance with USP <1790>.

[0255] In other embodiments, insoluble particulate matter with characteristic sizes from about 1 pm to about 100 pm that persist upon dissolution in an aqueous feed solution are referred to as Subvisible Particles ( SVPs). SVPs are present in quantities from about 0 to 100,000,000 per about 1 mL, e.g., from about 0 to about 10,000,000 per about 1 mL, from about 0 to about 1,000,000 per about 1 mL, from about 0 to about 500,000 per about 1 mL, from about 0 to about 100,000 per about 1 mL, from about 0 to about 50,000 per about 1 mL, from about 0 to about 10,000 per about 1 mL, from about 0 to about 6,000 per about 1 mL, from about 0 to about 1,000 per about 1 mL, from about 0 to about 600 per about 1 mL, from about 0 to about 250 per about 1 mL, from about 0 to about 100 per about 1 mL, from about 0 to about 60 per about 1 mL, or from about 0 to about 10 per about 1 mL. In other embodiments, the count of SVPs with characteristic size greater than or equal to 10 pm is from about 0 to about 6,000 per about 1 mL, e.g., from about 0 to about 1,000 per about 1 mL, from about 0 to about 100 per about 1 mL, from about 0 to about 10 per about 1 mL, from about 0 to about 5 per about 1 mL, from about 0 to about 3 per about 1 mL, or from about 0 to about 1 per about 1 mL. In certain embodiments, the count of SVPs with characteristic size greater than or equal to 25 pm is from about 0 to about 600 per about 1 mL, e.g., from about 0 to about 100 per about 1 mL, from about 0 to about 10 per about 1 mL, from about 0 to about 3 per about 1 mL, from about 0 to about 1 per about 1 mL, from about 0 to about 0.5 per about 1 mL, or from about 0 to about 0.1 per about 1 mL. Example methods of measuring SVPs include analysis of therapeutic agent with a Coulter Counter, HIAC Royco, or micro-flow imaging system after reconstitution and dilution of therapeutic agent to a standard concentration, e.g., about 100 mg / mL or about 1 mg / mL. In still other embodiments, the composition has a concentration of insoluble SVPs of about 0 per about 1 mL to about 100,000,000 per about 1 mL of greater than about 10 pm particles upon dissolution in an aqueous feed solution. In certain embodiments, the composition has a concentration of insoluble SVPs of about 0 per about 1 mL to about 6000 per about 1 mL of greater than about 10 pm particles upon dissolution in an aqueous feed solution. In embodiments, the composition has a concentration of insoluble SVPs of about 0 per about 1 mL to about 600 per about 1 mL of greater than about 25 pm particles upon dissolution in an aqueous feed solution. In certain embodiments, the composition is substantially free of insoluble SVPs upon dissolution in an aqueous feed solution. In 924248642. vl5726.1016001embodiments, the aqueous feed solution is water, aqueous buffer or a physiologically relevant aqueous feed solution.

[0256] In some embodiments, insoluble particulate matter with characteristic sizes from about 100 nm to about 1 pm that persist upon dissolution in an aqueous feed solution are referred to as submicron particles (SMP) and sometimes known as nanoparticles. Quantitatively, SMPs are present in quantities from about 0 to 5*1012per about 1 mL, e.g., from about 0 to about 0.5* 1012per about 1 mL, from about 0 to about 50* 109per about 1 mL, from about 0 to about lOMO9per about 1 mL, from about 0 to about 5 / I09per about 1 mL, from about 0 to about 0.5* 109per about 1 mL, from about 0 to about 50* 106per about 1 mL, from about 0 to about l><106per about 1 mL, from about 0 to about 500,000 per about 1 mL, from about 0 to about 200,000 per about 1 mL, from about 0 to about 100,000 per about 1 mL, from about 0 to about 10,000 per about 1 mL, from about 0 to about 5000 per about 1 mL, or from about 0 to about 1000 per about 1 mL. Example methods of measuring SMPs quantitatively include analysis of the therapeutic agent with a NanoSight, asymmetric field flow fractionation coupled to a multi-angle laser light scattering (AF4 MALS), Dynamic Light Scattering (DLS), or any other nano-particle tracking device known in the art, after reconstitution and dilution of the therapeutic agent to a standard concentration, e.g., about 100 mg / mL, about 1 mg / mL, or about 1 pg / mL. Qualitatively, SMPs are within a range comparable to the starting monomeric therapeutic agent solution. In embodiments, the composition is substantially free of submicron particles (SMP) upon dissolution in an aqueous feed solution. In certain embodiments, the aqueous feed solution is water, aqueous buffer or a physiologically relevant aqueous feed solution. In certain embodiments, the aqueous feed solution comprises a therapeutic agent and an excipient. In certain embodiments, the aqueous feed solution comprises a therapeutic agent and one or more excipients (e.g., two or more excipients). Qualitatively, as described herein, SMPs are within a range comparable to the feed solution.

[0257] In certain embodiments, the pharmaceutical composition (e.g., formulation) includes insoluble particulate matter smaller than or equal to 1 pm. The pharmaceutical composition can have a concentration of insoluble particles with a characteristic size greater than or equal to about 100 nm is about 1 to 5xl012per about 1 mL in suspension, or have a concentration of insoluble particles with a characteristic size less than or equal to about 1 pm is about 1 to 5xl012per about 1 mL in suspension. In still other embodiments, the pharmaceutical composition of particles may include insoluble particulate matter larger than or equal to about 1 pm in size. In certain other embodiments, the number of insoluble particles is from about 0 to about 100,000,000 per about 1 mL, e.g., less than about 10,000,000,934248642. vl5726.10160011,000,000, 100,000, 10,000, 1000, 100, 10, or about 1 per about 1 mL. For example, the number of insoluble particles greater than about 10 pm is from about 0 to about 6,000 per about 1 mL, e.g., less than about 5,000, about 4,000, about 3,000, about 2,000, about 1,000, about 500, about 100, about 10, or about 1 per about 1 mL, and / or the number of insoluble particles greater than about 25 pm is from about 0 to about 600 per about 1 mL, e.g., less than about 500, about 400, about 300, about 200, about 100, about 50, about 10, or about 1 about 1 per about 1 mL.

[0258] In some embodiments, the disclosure provides a pharmaceutical composition, e.g., a suspension or dried form, containing a plurality of particles that include a therapeutic agent, e.g., an antibody. The composition preferably has a concentration of insoluble particles, e.g., SVPs, of about 0 and about 100,000,000 per about 1 mL in suspension or upon reconstitution. In other embodiments, the concentration of insoluble particles is of about 0 and about 1,000,000 per about 1 mL in suspension or upon reconstitution. In still other embodiments, the concentration of insoluble particles is of about 0 and about 10,000 per about 1 mL in suspension or upon reconstitution. In certain other embodiments, the concentration of insoluble particles with a characteristic size greater than or equal to about 10 pm is of about 0 to about 6,000 per about 1 mL in suspension or upon reconstitution. In certain embodiments, the concentration of insoluble particles with a characteristic size greater than or equal to about 25 pm is of about 0 to about 600 per about 1 mL in suspension or upon reconstitution.

[0259] In other embodiments, after dissolution or reconstitution of the particles following storage, SVPs are present in quantities from about 0 to about 100,000,000 per about 1 mL, e.g., from about 0 to about 10,000,000 per about 1 mL, from about 0 to about 1,000,000 per about 1 mL, from about 0 to about 500,000 per about 1 mL, from about 0 to about 100,000 per about 1 mL, from about 0 to about 50,000 per about 1 mL, from about 0 to about 10,000 per about 1 mL, from about 0 to about 6,000 per about 1 mL, from about 0 to about 1,000 per about 1 mL, from about 0 to about 600 per about 1 mL, from about 0 to about 250 per about 1 mL, from about 0 to about 100 per about 1 mL, from about 0 to about 60 per about 1 mL, or from about 0 to about 10 per about 1 mL. In some embodiments, the count of particles with characteristic size greater than or equal to about 10 pm is from about 0 to about 6,000 per about 1 mL, e.g., from about 0 to about 1,000 per about 1 mL, from about 0 to about 100 per about 1 mL, from about 0 to about 10 per about 1 mL, from about 0 to about 5 per about 1 mL, from about 0 to about 3 per about 1 mL, or from about 0 to about 1 per about 1 mL. In certain embodiments, the count of particles with characteristic size greater than or equal to about 25 pm is from about 0 to about 600 per about 1 mL, e.g., from about 0 to about 100 per about 1 mL, from about 0 to about 10 per about 1 mL, from about 0 to about 3 per about 1 mL, from 944248642. vl5726.1016001about 0 to about 1 per about 1 mL, from about 0 to about 0.5 per about 1 mL, or from about 0 to about 0.1 per about 1 mL. In still other embodiments, after dissolution or reconstitution of the particles following storage, the therapeutic agent retains from about 0.5 to about 1.0 activity, e.g., from about 0.75 to about 1.0 activity, from about 0.9 to about 1.0 activity, from about 0.95 to about 1.0 activity, from about 0.99 to about 1.0 activity, or from about 0.999 to about 1.0 activity. In certain other embodiments, dissolution or reconstitution of the particles following storage provides less than about a 50% increase in aggregates of the therapeutic agent, e.g., an antibody, antibody fragment, or human serum albumin (HSA), (e.g., less than about 50%, less than about 25%, less than about 20%, less than about 15%, less than about 10%, less than about 8%, less than about 5%, less than about 4%, less than about 3%, less than about 1%, less than about 0.5%, or less than about 0.1%) as compared to the therapeutic agent in the aqueous liquid prior to processing. In certain embodiments, the dissolution or reconstitution of the particles after storage provides less than about a 50% increase in fragments of the therapeutic agent, e.g., an antibody, antibody fragment, or human serum albumin (HSA), (e.g., less than about 50%, less than about 25%, less than about 20%, less than about 15%, less than about 10%, less than about 8%, less than about 5%, less than about 4%, less than about 3%, less than about 1%, less than about 0.5%, or less than about 0.1%) as compared to the therapeutic agent in the aqueous liquid prior to processing. In some embodiments, the dissolution or reconstitution of the particles following storage provides less than about a 50% change in charge variants in the population of the therapeutic agent, e.g., an antibody or an antibody fragment, (e.g., less than about 40, 30, 20, 10, 8, 5, 4, 3, or about 1%) as compared to therapeutic agent prior to particle formation.

[0260] In some embodiments, after dissolution or reconstitution of the particles following storage, SVPs are present in quantities from about 0 to about 100,000,000 per about 1 mL, e.g., from about 0 to about 10,000,000 per about 1 mL, from about 0 to about 1,000,000 per about 1 mL, from about 0 to about 500,000 per about 1 mL, from about 0 to about 100,000 per about 1 mL, from about 0 to about 50,000 per about 1 mL, from about 0 to about 10,000 per about 1 mL, from about 0 to about 6,000 per about 1 mL, from about 0 to about 1,000 per about 1 mL, from about 0 to about 600 per about 1 mL, from about 0 to about 250 per about 1 mL, from about 0 to about 100 per about 1 mL, from about 0 to about 60 per about 1 mL, or from about 0 to about 10 per about 1 mL. In certain embodiments, the count of particles with characteristic size greater than or equal to about 10 pm is from about 0 to about 6,000 per about 1 mL, e.g., from about 0 to about 1,000 per about 1 mL, from about 0 to about 100 per about 1 mL, from about 0 to about 10 per about 1 mL, from about 0 to about 5 per 1 mL, from about 0954248642. vl5726.1016001to about 3 per about 1 mL, or from about 0 to about 1 per about 1 mL. In certain other embodiments, the count of particles with characteristic size greater than or equal to about 25 pm is from about 0 to about 600 per about 1 mL, e.g., from about 0 to about 100 per about 1 mL, from about 0 to about 10 per about 1 mL, from about 0 to about 3 per about 1 mL, from about 0 to about 1 per about 1 mL, from about 0 to about 0.5 per about 1 mL, or from about 0 to about 0.1 per about 1 mL.

[0261] Applicant herein discloses methods and systems for the formation of a suspension of particles including at least one therapeutic for administration to humans. The method and systems disclosed herein capture the surprising discovery of a method that avoids the introduction of SVPs. Applicant discloses that the methods described herein provide for the formation of droplets of an aqueous liquid, and subsequent production of particles and particle suspensions (including suspensions of particles), with minimal or no direct contact with mixing equipment having movable parts (e.g., an impeller or chopper) that can shed metal and other materials, which have been found to result in the introduction of SVPs into the collection of particles or particle compositions in undesirable amounts. The use of rotor-stator mixers, Tee-mixers, and static mixers as disclosed herein may allow for the controlled production of high concentration therapeutic suspensions meeting regulatory requirements for critical quality attributes (“CQAs”). Applicant discovered that producing particles and particle compositions for administration to humans by injection (e.g., subcutaneous or intramuscular) using the systems and methods disclosed herein produces a suspension likely meeting regulatory CQAs. Applicant’s methods and systems overcome the issues known in the art that drug products comprising particles of one or more therapeutic agents (e.g., a suspension of particles) cannot be filtered (i.e., terminal or sterile filtration) at the end of the manufacturing process to remove SVPs introduced during the process. Applicant discovered that the systems and methods disclosed herein limit or avoid direct contact with equipment having movable parts and achieve a 5%, 10%, 20%, 25%, 30%, 35%, 40%, 45%, 50% or greater reduction in SVPs compared to a process that employs equipment having movable parts, such as an impeller or chopper. Thereby allowing the formation of shelf-stable, high concentration therapeutic suspensions with low levels of SVPs for meeting regulator CQAs.

[0262] In some embodiments, dissolution or reconstitution of the particles following storage provides less than about a 50% increase in aggregates of the therapeutic agent, e.g., an antibody, antibody fragment, or human serum albumin (HSA), (e.g., less than about 45%, less than about 40%, less than about 35%, less than about 30%, less than about 25%, less than about 20%, less than about 15%, less than about 10%, less than about 8%, less than about 5%, less 964248642. vl5726.1016001than about 4%, less than about 3%, less than about 1%, less than about 0.5%, or less than about 0.1%) as compared to the therapeutic agent in the aqueous liquid prior to processing. In other embodiments, the dissolution or reconstitution of the particles after storage provides less than about a 50% increase in fragments of the therapeutic agent, e.g., an antibody, antibody fragment, or human serum albumin (HSA), (e.g., less than about 45%, less than about 40%, less than about 35%, less than about 30%, less than about 25%, less than about 20%, less than about 15%, less than about 10%, less than about 8%, less than about 5%, less than about 4%, less than about 3%, less than about 1%, less than about 0.5%, or less than about 0.1%) as compared to the therapeutic agent in the aqueous liquid prior to processing. In certain other embodiments, the dissolution or reconstitution of the particles following storage provides less than about 50% change in charge variants in the population of a therapeutic agent, e.g., an antibody, antibody fragment, or human serum albumin (HSA), e.g., less than about 40, about 30, about 20, about 10, about 8, about 5, about 4, about 3, or about 1%, as compared to the therapeutic agent prior to particle formation.

[0263] In certain embodiments, the present disclosure includes concentrated particles comprising at least one therapeutic agent, wherein the particles upon dissolution in water, buffers or other physiologically relevant aqueous feed solutions, e.g., biological fluids in the patients’ body, have a substantially similar turbidity compared to a composition comprising the therapeutic agent in the aqueous liquid. The term “turbidity” means the cloudiness or haziness of a fluid caused by individual particles that remain insoluble after dissolution at the desired concentration in water, buffer or other physiologically relevant aqueous feed solutions, e.g., biological fluids in the patients’ body. As used herein, “physiologically relevant” conditions as may be encountered inside a mammal or human, can apply. The skilled person will be able to determine the set of conditions most appropriate for testing in accordance with the ultimate application of the compositions described herein. In some embodiments, the particles upon dissolution in the aqueous liquid has a substantially similar turbidity compared to a composition comprising the therapeutic agent in the first aqueous feed solution. In embodiments, the particles upon dissolution in the aqueous liquid is substantially free of turbidity. In certain embodiments, the aqueous liquid is water, aqueous buffer or a physiologically relevant aqueous feed solution.

[0264] In some embodiments, the particles of the present disclosure can be reconstituted into a liquid pharmaceutical composition to assess the turbidity or turbidance (USP <855>). Turbidity may be measured in units of FTU (Formazin Turbidity Units). This is achieved by comparing the turbidity of a sample with that of a formazine suspension. Turbidity may also 974248642. vl5726.1016001be measured as Nephelometric Turbidity Units (NTU) where 1NTU = 1FTU. In other embodiments, when 10 mg of particles are dissolved in 1 mL of liquid, turbidity can be of about 0 to about 4000 FTU, about 0 to about 1000 FTU, about 0 to about 500 FTU, about 0 to about 50 FTU, about 0 to about 20 FTU, about 0 to about 10 FTU, about 0 to about 5 FTU, about 0 to about 1 FTU, about 0 to about 0.1 FTU, or about 0 to about 0.01 FTU. In certain embodiments, the pharmaceutical composition has a turbidity of about 0 to about 4000 Formazin Turbidity Units (FTU). In certain other embodiments, the pharmaceutical composition upon dissolution in an aqueous feed solution has a substantially similar turbidity compared to an aqueous composition comprising the therapeutic agent in monomeric form. In embodiments, the pharmaceutical composition upon dissolution in an aqueous feed solution is substantially free of turbidity. In certain embodiments, the aqueous feed solution is water, aqueous buffer, or a physiologically relevant aqueous feed solution.

[0265] In other embodiments, the present disclosure concerns highly concentrated compositions of low turbidity comprising a carbohydrate, a pH adjusting agent, a salt, a surfactant, a protein stabilizer, an emulsifier, an amino acid, a flocculation agent, and a plurality of particles comprising a therapeutic agent, in a non-aqueous carrier (e.g., propylene glycol diesters, ethyl oleate, medium chain triglycerides, triacetin, benzyl benzoate). In embodiments, the disclosure concerns highly concentrated pharmaceutical compositions of low turbidity comprising trehalose, arginine hydrochloride, sodium succinate, succinic acid, citric acid, sodium citrate, histidine, histidine hydrochloride, sodium chloride, hydroxypropyl betacyclodextrin, polysorbate, or sorbitan monooleate, and a plurality of particles comprising an antibody, in the non-aqueous carrier such as propylene glycol diesters, ethyl oleate, medium chain triglycerides, triacetin, or benzyl benzoate. In certain embodiments, the pharmaceutical composition upon dissolution in water, aqueous buffer or any physiologically relevant aqueous feed solution is substantially free of turbidity.

[0266] In some embodiments, droplets, particles, and / or compositions disclosed herein further comprise at least one hyaluronan degrading agent (e.g., the at least one hyaluronan degrading agent can be administered simultaneously, sequentially or intermittently with the composition). The subcutaneous tissue or the extracellular matrix is comprised of a network of fibrous proteins embedded within a viscoelastic gel of glycosaminoglycans. Hyaluronan is the prominent glycosaminoglycan of the subcutaneous tissue. Hyaluronan is secreted into the interstitium by fibroblasts as a viscous polymer that is subsequently degraded locally, in the lymph, through the action of hyaluronidases. Glycosaminoglycans are complex linear984248642. vl5726.1016001polysaccharides of the extracellular matrix and are characterized by repeating disaccharide structures of an N-substituted hexosamine and a uronic acid, as in the case of hyaluronan.

[0267] The hyaluronan degrading agent can enhance the subcutaneous administration of the composition comprising a plurality of particles and a flocculation agent, for example, by enhancing and / or increasing the volume of the composition being administered by injection thereby improving the absorption of the therapeutic agent. The use of a hyaluronan degrading agent, such as hyaluronidase, can improve the subcutaneous administration of the therapeutic agent into systemic circulation via the reversible hydrolyzation of hyaluronan, e.g., the reversible degradation of hyaluronan. The degradation of hyaluronan in the extracellular matrix temporarily opens channels in the subcutaneous tissue thereby allowing for larger volumes to be administered safely and comfortably into the subcutaneous tissue. Moreover, the degradation of hyaluronan temporarily decreases the viscosity of the subcutaneous tissue and promotes the dispersion of injected liquids facilitating their absorption. The effects of hyaluronidase are local and reversible with complete reconstitution of the hyaluronan tissue occurring within 24 to 48 hours. See, e.g., Frost, G. I., “Recombinant human hyaluronidase (rHuPH20): an enabling platform for subcutaneous drug and fluid administration”, Expert Opinion on Drug Delivery, 2007; 4:427-440], The increase in the permeability of the subcutaneous tissue through the degradation of hyaluronan correlates with the efficacy of hyaluronidase for their capability to increase the dispersion and absorption of compositions comprising a plurality of particles administered simultaneously, sequentially or intermittently. In certain embodiments, the methods described herein, further comprise administering at least one hyaluronan degrading agent. In certain embodiments, the hyaluronan degrading agent is a second therapeutic agent in the particles. In some embodiments, the hyaluronan degrading agent is in the suspension.

[0268] In certain embodiments, the hyaluronan degrading agent is a hyaluronidase enzyme, also referred to herein as a hyaluronidase. In some embodiments, the hyaluronidase is a soluble neutral-active hyaluronidase. In certain embodiments, the hyaluronidase is a mammalian hyaluronidase. In particular embodiments, the mammalian hyaluronidase is a human hyaluronidase. In particular embodiments, the human hyaluronidase is a recombinant human hyaluronidase, e.g., a rHuPH20. Certain recombinant human hyaluronidases that are suitable for use in the compositions disclosed herein are commercially available, e.g., rHuPH20 from Halozyme Therapeutics (San Diego, CA).

[0269] The particles comprising at least one therapeutic agent described herein, can be used in a number of ways, as well as any methods for the delivery of the particles disclosed in, for 994248642. vl5726.1016001example, in International Application Nos. PCT / US2023 / 066395 (Pub. No. WO 2023 / 212721), PCT / US2021 / 027755 (Pub. No. WO 2021 / 212019), PCT / US2017 / 063150 (Pub. No. WO 2018 / 098376), PCT / US2018 / 043774 (Pub. No. WO 2019 / 023392), PCT / US2019 / 033875 (Pub. No. WO 2019 / 226969), PCT / US2020 / 15957 (Pub. No. WO 2020 / 160323), PCT / US2020 / 050508 (Pub. No. WO 2021 / 050953), PCT / US2021 / 016878 (Pub. No.2021 / 158959), and PCT / US2021 / 018806 (Pub. No. WO 2021 / 168271), the contents of each of which are hereby incorporated by reference in their entireties.Post-Processing

[0270] In some embodiments, an organic liquid, organic solvent, or dehydration liquid used in any of the methods described herein is removed through filtration, solvent exchange, or a combination thereof. In some embodiments, an organic liquid, organic solvent, or dehydration liquid used in any of the methods described herein is removed using one or more hydrocyclones, continuous centrifugation, or one or more settling tanks. In some embodiments, the organic liquid is removed by filtration. In certain embodiments, the filtration comprises direct-flow filtration. In some embodiments, the filtration comprises using a porous filter. In some embodiments, the filter comprises sintered metal. In some embodiments, the dehydration liquid is an organic solvent.

[0271] In some embodiments, the filtration (e.g., direct-flow filtration) is performed using a filter pore size of from about 0.1 pm to about 100 pm (e.g., about 1 pm to about 90 pm, about 1 pm to about 80 pm, about 1 pm to about 70 pm, about 1 pm to about 60 pm, about 1 pm to about 50 pm, about 1 pm to about 40 pm, about 1 pm to about 30 pm, about 1 pm to about 20 pm, about 1 pm to about 10 pm, about 1 pm to about 5 pm, about 0.1 pm to about 9 pm, about 0.1 pm to about 8 pm, about 0.1 pm to about 7 pm, about 0.1 pm to about 6 pm, about 0.1 pm to about 5 pm, about 0.1 pm to about 4 pm, about 0.1 pm to about 3 pm, about 0.1 pm to about 2 pm, about 0.1 pm to about 1 pm, etc.). In some embodiments, the filtration is performed using a filter pore size of from about 1 pm to about 50 pm. In some embodiments, the filter pore size is about 100 pm, about 75 pm, about 50 pm, about 25 pm, about 15 pm, about 10 pm, about 5 pm. In some embodiments, the filter is configured to retain a plurality of particles with a volume-weighted average particle size distribution.

[0272] In some embodiments, the direct-flow filtration is performed using a flow rate of from about 0.1 L / min to about 100 L / min (c.g, about 0.1 mL / min to about 90 mL / min, about 0.1 mL / min to about 80 mL / min, about 0.1 mL / min to about 70 mL / min, about 0.1 mL / min to about 60 mL / min, about 0.1 mL / min to about 50 mL / min, about 0.1 mL / min to about 401004248642. vl5726.1016001mL / min, about 0.1 mL / min to about 30 mL / min, about 0.1 mL / min to about 20 mL / min, about 0.1 mL / min to about 10 mL / min, about 0.5 mL / min to about 10 mL / min, about 1 mL / min to about 10 mL / min, etc.). In some embodiments, the direct-flow filtration is performed using a flow rate of from about 0.1 L / min to about 50 L / min. In some embodiments, the flow rate is from about 1 L / min to about 10 L / min.

[0273] In some embodiments, the direct-flow filtration is performed using a filter loading of from about 0.1 kg / m2to about 50 kg / m2(e.g., about 0.5 kg / m2to about 50 kg / m2, about 1 kg / m2to about 50 kg / m2, about 1 kg / m2to about 40 kg / m2, about 1 kg / m2to about 30 kg / m2, about 1 kg / m2to about 20 kg / m2, about 1 kg / m2to about 10 kg / m2, about 1 kg / m2to about 9 kg / m2, about 1 kg / m2to about 8 kg / m2, about 1 kg / m2to about 7 kg / m2, about 1 kg / m2to about 6 kg / m2, about 1 kg / m2to about 5 kg / m2, about 1 kg / m2to about 4 kg / m2, about 1 kg / m2to about 3 kg / m2, about 1 kg / m2to about 2 kg / m2, etc.). In some embodiments, the direct-flow filtration is performed using a filter loading of from about 1 kg / m2to about 2 kg / m2. In some embodiments, the filter loading is about 1.8 kg / m2.

[0274] In some embodiments, the direct-flow filtration is performed using a flux of from about 100 L / m2h to about 10000 L / m2h (e.g., about 100 L / m2h to about 9000 L / m2h, about 100 L / m2h to about 8000 L / m2h, about 100 L / m2h to about 7000 L / m2h, about 100 L / m2h to about 6000 L / m2h, about 100 L / m2h to about 5000 L / m2h, about 500 L / m2h to about 5000 L / m2h, about 600 L / m2h to about 5000 L / m2h, about 700 L / m2h to about 5000 L / m2h, about 800 L / m2h to about 5000 L / m2h, about 900 L / m2h to about 5000 L / m2h, about 1000 L / m2h to about 5000 L / m2h, about 1000 L / m2h to about 4000 L / m2h, about 2000 L / m2h to about 4000 L / m2h, about 3000 L / m2h to about 4000 L / m2h, etc.). In some embodiments, the direct-flow filtration is performed using a flux of from about 100 L / m2h to about 10000 L / m2h. In some embodiments, the flux is from about 1000 L / m2h to about 5000 L / m2h.

[0275] In other embodiments, the methods as described herein, further comprise washing the particles with a washing fluid, e.g., an organic liquid, an organic solvent, a dehydration liquid, a supercritical fluid, a cryogenic liquid, or a combination thereof. In certain embodiments, the washing fluid is an organic liquid, an organic solvent, a dehydration liquid, a supercritical fluid, a cryogenic liquid, a carrier liquid, or a combination thereof. In some embodiments, the particles may be washed with the washing fluid at least once to remove residual organic liquid, organic solvent, dehydration liquid, and / or aqueous feed solution from the particles. In certain embodiments, the particles may be washed 1, 2, 3, 4, or 5 times to remove residual organic liquid, organic solvent, dehydration liquid, and / or a residual aqueous...

Claims

1. 5726.10160012.CLAIMS3.What is claimed is:

1. A method of making a suspension of particles comprising a therapeutic agent, the method comprising:5.a) providing a first liquid stream comprising an aqueous feed solution, the feed solution comprising the therapeutic agent;6.b) providing a second liquid stream comprising an organic liquid;7.c) dispersing the first liquid stream into the second liquid stream at a junction; d) applying turbulent flow conditions to a fluid channel connected to the junction, the fluid channel containing the first liquid stream and the second liquid stream, to form aqueous droplets comprising the therapeutic agent;8.e) dehydrating an aqueous fraction of the aqueous droplets, the dehydrating comprising contacting the aqueous droplets with a dehydration liquid to form particles comprising the therapeutic agent;9.f) removing the dehydration liquid; and10.g) suspending the particles in a pharmaceutically acceptable carrier liquid to form the suspension.

2. The method of claim 1, wherein the suspension has a water content of less than about 20% (w / v).

3. The method of claim 1 or 2, wherein the therapeutic agent is a therapeutic biologic or a salt thereof.

4. The method of any one of claims 1-3, wherein the therapeutic agent is an antibody or a fragment thereof.

5. The method of any one of claims 1-4, wherein the therapeutic agent has a concentration of from about 70 g / L to about 160 g / L in the first liquid stream.

6. The method of any one of claims 1-5, wherein the first liquid stream further comprises one or more excipients.16.1274248642. vl5726.10160017. The method of claim 6, wherein the one or more excipients comprises a surfactant, an amino acid, a carbohydrate, a salt, an antioxidant, or a combination thereof.

8. The method of any one of claims 1-7, wherein the organic liquid is an ester or an alcohol.

9. The method of any one of claims 1-8, wherein the organic liquid is n-butyl acetate or pentanol.

10. The method of any one of claims 1-9, wherein the first liquid stream flow rate is from about 10 mL / min to about 100 mL / min.

11. The method of claim 10, wherein the first liquid stream flow rate is from about 20 mL / min to about 30 mL / min.

12. The method of any one of claims 1-11, wherein the second liquid stream flow rate is a turbulent flow rate from about 0.1 L / min to about 50 L / min.

13. The method of claim 12, wherein the second liquid stream flow rate is from about 1 L / min to about 20 L / min.

14. The method of any one of claims 1-13, wherein the second liquid stream flow rate to first liquid stream flow rate ratio is from about 1 : 1 to about 500: 1.

15. The method of claim 14, wherein the flow conditions within the fluid channel are highly turbulent.

16. The method of any one of claims 1-15, wherein the first liquid stream is dispersed into the second liquid stream at a temperature of from about 3°C to about 40°C.

17. The method of any one of claims 1-16, wherein the first liquid stream is dispersed into a second liquid stream using a rotor stator mixer, Tee-mixer, or junction with static mixer.29.1284248642. vl5726.101600118. The method of claim 17, wherein the first liquid stream is dispersed into a second liquid stream using one or more Tee-mixers or one or more junctions with static mixers.

19. The method of any one of claims 1-18, wherein dehydrating an aqueous fraction of the aqueous droplets comprises contacting the aqueous droplets with a dehydration liquid in a drying tube, a drying vessel, or a combination thereof.

20. The method of claim 19, wherein the drying tube is a dehydration coil.

21. The method of claim 19, wherein the residence time of the aqueous droplets in the drying tube or vessel is at least about 10 seconds.

22. The method of claim 19, wherein the residence time of the aqueous droplets in the drying tube or vessel is at least about 15 seconds.

23. The method of any one of claims 1-22, wherein the organic liquid is the same liquid as the dehydration liquid.

24. The method of any one of claims 1-23, wherein a plurality of the particles formed in e) have a number-weighted average circularity of from about 0.80 to about 1.00.

25. The method of any one of claims 1-24, wherein the dehydration liquid in f) is removed through filtration, solvent exchange, or a combination thereof.

26. The method of any one of claims 1-25, wherein the dehydration liquid in f) is removed using one or more hydrocyclones, continuous centrifugation, or one or more settling tanks.

27. The method of claim 26, wherein the filtration comprises using a porous filter.

28. The method of claim 27, wherein the filtration is performed using a filter pore size of from about 0.1 pm to about 5 pm.42.1294248642. vl5726.101600129. The method of claim 27 or 28, wherein the filter comprises sintered metal.

30. The method of any one of claims 25 and 27-29, wherein the filtration comprises performing direct-flow filtration.

31. The method of claim 30, wherein the direct-flow filtration is performed using a flow rate of from about 0.1 L / min to about 50 L / min.

32. The method of claim 31, wherein the direct-flow filtration flow rate is from about 1 L / min to about 20 L / min.

33. The method of any one of claims 27-29, wherein the filter is configured to retain a plurality of particles with a volume-weighted average particle size distribution.

34. The method of any one of claims 30-32, wherein the direct-flow filtration is performed using a flux of from about 100 L / m2h to about 10000 L / m2h.

35. The method of claim 34, wherein the flux is from about 1000 L / m2h to about 5000 L / m2h.

36. The method of any one of claims 27-35, wherein the filter pore size is less than about 5 pm.

37. The method of any one of claims 1-36, wherein step f) comprises exchanging the dehydration liquid for a pharmaceutically acceptable carrier liquid without forming a powder intermediate.

38. The method of claim 37, wherein the pharmaceutically acceptable carrier liquid comprises benzyl benzoate, medium chain triglycerides, propylene glycol diesters, triacetin or ethyl oleate.54.1304248642. vl5726.101600139. The method of claim 37, wherein step f) comprises forming a powder intermediate and resuspending the powder intermediate in a pharmaceutically acceptable carrier liquid.

40. The method of any one of claims 37-39, wherein step f) is performed using a pharmaceutically acceptable carrier liquid flow rate of from about 100 mL / min to about 300 mL / min.

41. The method of any one of claims 37-40, wherein step f) is performed using a discharge pressure of from about 25 psi to about 30 psi.

42. The method of any one of claims 37-41, wherein step f) is performed using a discharge time of from about 10 s to about 100 s.

43. The method of claim 42, wherein the discharge time is from about 50 s to about 100 s.

44. The method of any one of claims 1-43, further comprising removing residual dehydration liquid and / or residual water from the suspension of particles in the pharmaceutically acceptable carrier liquid.

45. The method of any one of claims 1-43, further comprising performing sparging on the suspension of particles in the pharmaceutically acceptable carrier liquid.

46. The method of claim 45, wherein the sparging is performed using a gas flow rate of from about 0.1 L / min to about 100 L / min.

47. The method of claim 46, wherein the gas flow rate is from about 1 L / min to about 100 L / min.

48. The method of any one of claims 45-47, wherein the sparging is performed using nitrogen gas.

49. The method of any one of claims 45-48, wherein the sparging is performed using a gas relative humidity percentage of from about 0.1% to about 50%.67.1314248642. vl5726.101600150. The method of claim 49, wherein the gas relative humidity percentage is from about 1% to about 10%.

51. The method of any one of claims 45-50, wherein the sparging is performed using a mixing speed of from about 10 rpm to about 1000 rpm.

52. The method of claim 51, wherein the mixing speed is from about 100 rpm to about 500 rpm.

53. The method of any one of claims 45-52, wherein the sparging is performed for at least about 4 hours.

54. The method of claim 53, wherein the sparging is performed for about 8 hours.

55. The method of any one of claims 45-54, wherein the sparging is performed at a temperature of from about 20°C to about 30°C.

56. The method of any one of claim 1-55, the method further comprising removing at least a plurality of particles or particle aggregates of greater than about 50 pm in diameter.

57. The method of claim 56, wherein the removing comprises using sieving, cross-flow filtration, or a combination thereof.

58. The method of claim 57, wherein the sieving is performed using a sieving pressure of from about 1 psi to about 100 psi.

59. The method of claim 58, wherein the sieving pressure is from about 1 psi to about 50 psi.

60. The method of any one of claims 57-59, wherein the sieving is performed from about 1 min to about 100 min.80.1324248642. vl5726.101600161. The method of claim 60, wherein the sieving is performed from about 2 min to about 10 min.

62. The method of any one of claims 57-61, wherein the sieving is performed using a sieve loading of less than about 12 kg / m2.

63. The method of any one of claims 57-62, wherein the sieving is performed using a unidirectional sieve.

64. The method of claim 63, wherein the sieve removes a plurality of particles with a volume-weighted average diameter greater than 100 pm.

65. The method of any one of claims 1-64, the method further comprising concentrating the particles, the concentrating comprising performing cross-flow filtration.

66. The method of claim 65, wherein the cross-flow filtration is performed using a feed flow rate of from about 100 mL / min to about 5,000 mL / min.

67. The method of claims 1-66, the method further comprising concentrating the particles after suspending the particles in the pharmaceutically acceptable carrier liquid using a process that comprises cross-flow filtration.

68. The method of claim 66, wherein the feed flow rate is from about 1000 mL / min to about 2000 mL / min.

69. The method of any one of claims 66-68, wherein the cross-flow filtration is performed using a transmembrane pressure of from about 10 psi to about 15 psi.

70. The method of any one of claims 66-69, wherein the cross-flow filtration is performed using a membrane loading of from about 500 to about 1000 g / m2.

71. The method of any one of claims 1-70, wherein prior to a), the aqueous feed solution is warmed for at least about an hour.93.1334248642. vl5726.101600172. The method of claim 71, wherein the aqueous feed solution is warmed to a temperature of from about 15°C to about 30°C.

73. The method of any one of claims 1-72, the method further comprising adding the particles to a container.

74. The method of claim 73, wherein the container is a vial, cartridge, or syringe.

75. A method of making a suspension of particles comprising at least one therapeutic agent, the method comprising:99.a) providing a first liquid stream comprising an aqueous feed solution, the feed solution comprising the therapeutic agent;100.b) providing a second liquid stream comprising an organic liquid;101.c) dispersing the first liquid stream into the second liquid stream at a junction; d) applying turbulent flow conditions to a fluid channel connected to the junction, the fluid channel containing the first liquid stream and the second liquid stream, to form aqueous droplets comprising the therapeutic agent; e) dehydrating an aqueous fraction of the aqueous droplets, the dehydrating comprising contacting the droplets with a dehydration liquid to form particles comprising the therapeutic agent; and102.f) suspending the particles in a carrier liquid to form the suspension of particles aseptically.

76. The method of claim 75, wherein the fluid channel is coupled to a second junction.

77. The method of claim 76, wherein the second junction comprises a rotor stator mixer, Tee-mixer, or static mixer.

78. The method of claim 76, wherein the first liquid stream has a feed flow rate of from about 10 mL / min to about 50 mL / min.

79. The method of any one of claims 78, wherein the second liquid stream has a turbulent liquid flow rate of from about 0.1 L / min to about 50 L / min.107.1344248642. vl5726.101600180. The method of claim 79, wherein the turbulent liquid flow rate is from about 1 L / min to about 10 L / min.

81. The method of any one of claims 75, wherein the turbulent flow conditions are applied using a first liquid stream flow rate to second liquid stream flow rate ratio of from about 1 : 1 to about 500: 1.

82. The method of claim 81, wherein the second liquid stream flow rate to first liquid stream flow rate ratio is about 10: 1 to about 500: 1.

83. The method of any one of claims 75-82, wherein the turbulent flow conditions are applied at a temperature of from about 18°C to about 22°C.

84. The method of any one of claims 75-83, wherein step f) comprises exchanging the dehydration liquid with the carrier liquid without forming a powder intermediate.

85. The method of any one of claims 75-84, wherein the organic liquid is an ester or an alcohol.

86. The method of claim 85, wherein the organic liquid is n-butyl acetate or pentanol.

87. The method of any one of claims 75-86, wherein the therapeutic agent is a therapeutic biologic or salt.

88. The method of any one of claims 75-87, wherein the therapeutic agent is an antibody or a fragment thereof.

89. The method of any one of claims 75-88, wherein the first liquid stream further comprises one or more excipients.

90. The method of any one of claims 75-89, wherein the therapeutic agent has a concentration of from about 70 g / L to about 160 g / L in the first liquid stream.120.1354248642. vl5726.101600191. The method of claim 89 or 90, wherein the one or more excipients comprises a surfactant, an amino acid, a carbohydrate, a salt, an antioxidant, or a combination thereof.

92. A method of adjusting the water content of a plurality of particles in a suspension, the method comprising performing sparging on the suspension, wherein the suspension comprises the particles suspended in a carrier liquid, and wherein the plurality of the particles comprise at least one therapeutic agent.

93. The method of claim 92, wherein the sparging is performed using nitrogen gas.

94. The method of claim 92 or 93, wherein the sparging is performed using a gas relative humidity percentage of from about 0.1% to about 50%.

95. The method of claim 94, wherein the gas relative humidity percentage is from about 1% to about 10%.

96. The method of any one of claims 92-95, wherein the sparging is performed using a mixing speed of from about 10 rpm to about 1000 rpm.

97. The method of claim 96, wherein the mixing speed is from about 100 rpm to about 500 rpm.

98. The method of any one of claims 92-97, wherein the sparging is performed for at least about 4 hours.

99. The method of claim 98, wherein the sparging is performed for about 8 hours.

100. The method of any one of claims 92-99, wherein the sparging is performed at a temperature of from about 20°C to about 25°C.

101. The method of any one of claims 92-100, wherein the carrier liquid comprises benzyl benzoate, medium chain triglycerides, propylene glycol diesters, triacetin, or ethyl oleate.133.1364248642. vl5726.1016001102. The method of claim 101, wherein the carrier liquid comprises propylene glycol diesters.

103. The method of any one of claims 92-102, wherein the therapeutic agent is a therapeutic biologic or a salt thereof.

104. The method of any one of claims 92-103, wherein the therapeutic agent is an antibody or a fragment thereof.

105. The method of any one of claims 1-91, wherein the particles have a therapeutic agent density of about 1.0-3.0 g / cm3.

106. The method of any one of claims 1-91 and 105, further comprising adding a flocculation agent to the suspension.

107. The method of any one of claims 1-91 105, and 106, wherein the therapeutic agent has a concentration of from about 70 g / L to about 160 g / L in the first liquid stream.

108. The method of any one of claims 1-91, and 105-107, further comprising filtering at least a plurality of particles in the suspension through a filter having a pore size of at least about 75 pm.

109. The method of any one of claims 1-91 and 105-107, further comprising filtering at least a portion of particles in the suspension through a filter having a pore size of at most 75 pm.

110. The method of any one of claims 1-91 and 105-108, further comprising filtering at least a portion of particles in the suspension through a filter having a pore size of about 100 pm.

111. The method of any one of claims 1-91 and 105-110, further comprising filtering at least a portion of particles in the suspension through a filter having a pore size of about 75 pm.145.1374248642. vl5726.1016001112. The method of any one of claims 1-91, 105-107 and 109, further comprising filtering at least a portion of particles in the suspension through a filter having a pore size of about 50 pm.

113. The method of any one of claims 92-104, wherein the plurality of particles further comprises an excipient.

114. The method of claim 113, wherein the excipient comprises a surfactant, an amino acid, a carbohydrate, a salt, an antioxidant, or a combination thereof.

115. The method of any one of claims 89-91, wherein each of the first liquid stream and the particles further comprise two or more excipients, wherein each of the two or more excipients comprises a surfactant, an amino acid, a carbohydrate, a salt, an antioxidant, or a combination thereof.

116. The method of any one of claims 1-91, further comprising retaining the suspension in a fill-finish vessel.

117. The method of claim 116, further comprising pumping the suspension from the fillfinish vessel into at least one vial, cartridge, or syringe.

118. The method of any one of claims 1-91, further comprising quantifying a concentration of the particles in the suspension using nuclear magnetic resonance.

119. The method of claim 118, wherein the nuclear magnetic resonance quantifies the concentration of the particles in the suspension within the at least one vial, cartridge, or pre-filled syringe.

120. The method of claim 116, further comprising quantifying a concentration of the particles in the fill / finish vessel using nuclear magnetic resonance.156.1384248642. vl5726.1016001121. The method of claim 117, further comprising quantifying a concentration of the particles in the suspension pumped from the fill-finish vessel into at least one syringe using nuclear magnetic resonance.

122. The method of any one of claims 118-121, wherein quantifying the concentration of the particles comprises obtaining the relaxation rate of the suspension.

123. The method of any one of claims 118-122, wherein the quantifying further comprises obtaining the relaxation rate of the carrier liquid.

124. The method of any one of claims 118-123, wherein the quantifying further comprises obtaining the relaxation number from the relaxation rate of the suspension and the relaxation rate of the carrier liquid.

125. The method of any one of claims 1-124, wherein the suspension comprises a plurality of particles having a volume-weighted average diameter between about 5 pm and about 50 pm.

126. The method of any one of claims 1-125, wherein the suspension comprises a plurality of particles having a volume-weighted average diameter between about 10 pm and about 30 pm.

127. The method of any one of claims 1-91, wherein the method is an aseptic process performed in a closed system without forming a mostly dry powder intermediate.

128. A method of forming particles comprising a therapeutic agent, the method comprising:166.a) providing a first liquid stream comprising an aqueous feed solution, the feed solution comprising the therapeutic agent;167.b) providing a second liquid stream comprising an organic liquid;168.c) dispersing the first liquid stream into the second liquid stream at a first junction;169.d) applying turbulent flow conditions to a fluid channel connected to the first junction and a second junction, the fluid channel containing the first liquid170.1394248642. vl5726.1016001172.stream and the second liquid stream, to form aqueous droplets comprising the therapeutic agent;173.e) dehydrating an aqueous fraction of the aqueous droplets, the dehydrating comprising contacting the droplets with a dehydration liquid to form particles comprising the therapeutic agent; and174.f) collecting the particles aseptically.

129. The method of claim 128, wherein step f) comprises collecting the particles on a membrane.

130. The method of claim 128 or 129, wherein the first liquid stream further comprises an excipient.

131. A method of making a suspension of particles comprising a therapeutic agent, the method comprising:178.a) providing a first liquid stream comprising an aqueous feed solution, the feed solution comprising the therapeutic agent;179.b) providing a second liquid stream comprising an organic liquid;180.c) dispersing the first liquid stream into the second liquid stream at a first junction;181.d) applying turbulent flow conditions to a fluid channel connected to the first junction and a second junction, the fluid channel containing the first liquid stream and the second liquid stream, to form aqueous droplets comprising the therapeutic agent;182.e) dehydrating an aqueous fraction of the aqueous droplets, the dehydrating comprising contacting the droplets with a dehydration liquid to form particles comprising the therapeutic agent;183.f) exchanging the dehydration liquid with a carrier liquid aseptically to form the suspension of particles; and184.g) sparging the suspension of particles to reach a predetermined volume- weighted average water content in the particles.

132. The method of claim 131, wherein the volume-weighted average water content in the particles is between about 5% and about 20% (w / v).186.1404248642. vl5726.1016001133. The method of claim 131 or 132, the method further comprising: h) filtering out a portion of the particles in the suspension of particles, and wherein the volume- weighted average water content in the particles after step h) is between about 0% and about 10% (w / v).

134. The method of any one of claims 131-133, wherein step g) comprises suspending the particles directly into the carrier liquid without forming a powder intermediate.

135. The method of any one of claims 131-134, wherein step a) comprises performing tangential flow filtration to exchange an initial excipient profile of the first liquid stream with a second excipient profile.

136. The method of any one of claims 131-135, wherein the carrier liquid is a pharmaceutically acceptable carrier liquid.

137. The method of any one of claims 128-136, wherein the therapeutic agent is a therapeutic biologic or a salt thereof.

138. The method of any one of claims 128-137, wherein the therapeutic agent is an antibody or a fragment thereof.

139. The method of any one of claims 128-138, wherein the therapeutic agent has a concentration of from about 70 g / L to about 160 g / L in the first liquid stream.

140. The method of any one of claims 128-139, wherein the first liquid stream further comprises an excipient.

141. The method of claim 140, wherein the excipient comprises a surfactant, an amino acid, a carbohydrate, a salt, an antioxidant, or a combination thereof.

142. The method of any one of claims 128-141, wherein the organic liquid and / or dehydration liquid is an ester or an alcohol.198.1414248642. vl5726.1016001143. The method of any one of claims 128-142, wherein the organic liquid and / or dehydration liquid is n-butyl acetate or pentanol.

144. The method of any one of claims 128-143, wherein the first liquid stream is dispersed into a second liquid stream using a rotor stator mixer, Tee-mixer, or junction with static mixer.

145. The method of any one of claims 144, wherein the first liquid stream is dispersed into a second liquid stream using one or more Tee-mixers or one or more junctions with static mixers.

146. The method of any one of claims 128-145, wherein the dispersing is performed using a first liquid stream flow rate of from about 10 mL / min to about 50 mL / min.

147. The method of claim 146, wherein the first liquid stream flow rate is from about 20 mL / min to about 30 mL / min.

148. The method of any one of claims 128-147, wherein the dispersing is performed using a second liquid stream flow rate of from about 0.1 L / min to about 50 L / min.

149. The method of claim 148, wherein the second liquid stream flow rate is from about 1 L / min to about 10 L / min.

150. The method of any one of claims 128-149, wherein the dispersing is performed using a first liquid stream flow rate to second liquid stream flow rate ratio of from about 1:10 to about 1:500.

151. The method of claims 128-150, wherein the dispersing is performed using a first liquid stream flow rate to second liquid stream flow rate ratio of from about 10: 1 to about 500:1.

152. The method of any one of claims 128-151, wherein the dispersing is performed at a temperature of from about 18°C to about 22°C.210.1424248642. vl5726.1016001153. The method of any one of claims 128-152, wherein step d) applies turbulent flow conditions using a fitting, a rotor stator mixer, a Tee-mixer, or a static mixer at the first junction and / or the second junction.

154. The method of any one of claims 128-153, wherein the dehydrating occurs in a drying tube, a drying vessel, or a combination thereof.

155. The method of claim 154, wherein the drying tube is a dehydration coil.

156. The method of claim 154, wherein the residence time of the aqueous droplets in the drying tube, drying vessel or combination thereof is at least about 10 seconds.

157. The method of claim 156, wherein the residence time of the aqueous droplets in the drying tube, drying vessel, or combination thereof is about 15 seconds.

158. The method of any one of claims 128-157, wherein the number-weighted average circularity of at least a plurality of the particles is from about 0.80 to about 1.00.

159. The method of any one of claims 131-158, wherein the sparging is performed using a gas flow rate of from about 0.1 L / min to about 100 L / min.

160. The method of claim 159, wherein the gas flow rate is from about 1 L / min to about 100 L / min.

161. The method of any one of claims 131-160, wherein the sparging is performed using nitrogen gas.

162. The method of any one of claims 131-161, wherein the sparging is performed using a gas relative humidity percentage of from about 0.1% to about 50%.

163. The method of claim 162, wherein the gas relative humidity percentage is from about 1% to about 10%.223.1434248642. vl5726.1016001164. The method of any one of claims 131-163, wherein the sparging is performed using a mixing speed of from about 10 rpm to about 1000 rpm.

165. The method of claim 164, wherein the mixing speed is from about 100 rpm to about 500 rpm.

166. The method of any one of claims 131-165, wherein the sparging is performed for at least about 4 hours.

167. The method of claim 166, wherein the sparging is performed for about 8 hours.

168. The method of any one of claims 131-167, wherein the sparging is performed at a temperature of from about 20°C to about 25°C.

169. The method of any one of claims 128-168, the method further comprising removing particles or particle aggregates having a volume-weighted average diameter of greater than about 50 pm.

170. The method of claim 169, wherein the removing comprises using sieving, cross-flow filtration, or a combination thereof.

171. The method of claim 170, wherein the sieving is performed using a sieving pressure of from about 1 psi to about 100 psi.

172. The method of claim 171, wherein the sieving pressure is from about 1 psi to about 50 psi.

173. The method of any one of claims 170-172, wherein the sieving is performed from about 1 min to about 100 min.

174. The method of claim 173, wherein the sieving is performed from about 2 min to about 10 min.236.1444248642. vl5726.1016001175. The method of any one of claims 170-174, wherein the sieving is performed using a sieve loading of less than about 12 kg / m2.

176. The method of any one of claims 170-175, wherein the sieving is performed using a unidirectional sieve.

177. The method of claim 176, wherein the sieve filters out a plurality of particles with a volume-weighted average diameter of greater than about 100 pm.

178. A method of forming particles comprising a therapeutic agent, the method comprising:242.a) providing a first liquid stream comprising an aqueous feed solution comprising the therapeutic agent;243.b) providing a second liquid stream comprising an organic liquid;244.c) dispersing the first liquid stream into the second liquid stream at or after a first junction;245.d) applying turbulent flow conditions to a fluid channel connected to the first junction and a second, the fluid channel containing the first liquid stream and the second liquid stream, to form aqueous droplets comprising the therapeutic agent;246.e) dehydrating an aqueous fraction of the aqueous droplets, the dehydrating comprising contacting the droplets with a dehydration liquid at a second junction to form particles comprising the therapeutic agent, wherein the Peclet number of the first liquid stream, the second liquid stream and the dehydration liquid determines the morphology of the particles; and247.f) collecting the particles, thereby forming particles comprising the therapeutic agent, wherein the particles have a circularity of from about 0.80 to about 1.00.248.1454248642. vl

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