Compositions and methods for delivery of therapeutic biological agents for the treatment of disease

Abstract

To provide methods and compositions that allow for the delivery (e.g., subcutaneous delivery) of biopharmaceutical products for therapy.SOLUTION: A method useful for treating a disease or condition in a subject in need thereof, comprising administering to the subject a pharmaceutically effective amount of a plurality of particles suspended in a pharmaceutically acceptable liquid carrier of low viscosity, wherein: Wherein each of the particles comprises at least one therapeutic biological agent or a salt thereof; wherein the particles have less than about 10% aggregates of the therapeutic biological agent or salt thereof; and wherein the concentration of the therapeutic biological agent or salt thereof in the composition is from about 20 mg / ml to about 700 mg / ml.SELECTED DRAWING: None

Description

[Technical Field]

[0001] Related Applications This application claims the benefit of U.S. Patent Application No. 63 / 024,703, filed May 14, 2020, U.S. Patent Application No. 62 / 899,907, filed September 13, 2019, and U.S. Patent Application No. 62 / 899,981, filed September 13, 2019. The entire teachings of the above applications are incorporated herein by reference.

[0002] government support This invention was made with government support under Grant No. 1831212 awarded by the National Science Foundation. The government has certain rights in this invention.

[0003] The present disclosure relates to methods and compositions that enable the delivery (e.g., subcutaneous delivery) of biopharmaceutical products for therapy. In particular, the delivery methods disclosed herein use high-concentration, low-volume, and low-viscosity compositions of biological agents that allow for a practical transition from intravenous to subcutaneous delivery of therapeutic agents, and from frequent to less frequent subcutaneous delivery. [Background technology]

[0004] Biologic agents, particularly antibodies, have driven a paradigm shift in drug discovery and development over the past few decades, helping patients for whom few or no treatment options previously existed. For example, current monoclonal antibody (mAb) therapies often require large doses, which are administered via intravenous (IV) infusion at high volumes and low concentrations, which can take several hours to deliver, resulting in patient discomfort and increased risk of infection. Subcutaneous (SC) injections offer a more desirable alternative for delivery because they reduce the burden on hospitals and clinical facilities, require less time, and lower the risk of complications. However, SC injections require low delivery volumes, which necessitate high therapeutic biologic agent concentrations that are often difficult to achieve. The need for high concentrations at low delivery volumes also results in highly viscous injection solutions, which can lead to excessively high injection forces. Furthermore, highly viscous injection solutions of therapeutic biologic agents, such as mAbs, can cause increased protein-protein interactions, resulting in product loss and impacting product safety. Therefore, there is a need for highly concentrated, low-volume, low-viscosity injectable solutions for SC delivery of therapeutic biopharmaceutical products. Summary of the Invention [Means for solving the problem]

[0005] Provided herein are methods useful for treating a disease or condition in a subject in need thereof, comprising administering a pharmaceutically effective amount of: A plurality of particles suspended in a low viscosity pharmaceutically acceptable liquid carrier, each particle comprising at least one therapeutic biological agent or salt thereof. administering to a subject a composition comprising: the particles have less than about 10% aggregates of the therapeutic biological agent or salt thereof; and The concentration of the therapeutic biological agent or salt thereof in the composition is from about 20 mg / mL to about 700 mg / mL.

[0006] In one aspect, the disclosure provides a method of treating cancer in a subject in need thereof. That is, a pharmaceutically effective amount of A plurality of particles suspended in a low viscosity pharmaceutically acceptable liquid carrier, each particle comprising at least one therapeutic biological agent or salt thereof. administering to a subject a composition comprising: the particles have less than about 10% aggregates of the therapeutic biological agent or salt thereof; and The method provides a method in which the concentration of the therapeutic biological agent or salt thereof in the composition is from about 20 mg / mL to about 700 mg / mL.

[0007] The disclosure herein provides a method of treating an inflammatory disease or condition in a subject in need thereof, comprising administering a pharmaceutically effective amount of: A plurality of particles suspended in a low viscosity pharmaceutically acceptable liquid carrier, each particle comprising at least one therapeutic biological agent or salt thereof. administering to a subject a composition comprising: the particles have less than about 10% aggregates of the therapeutic biological agent or salt thereof; and Also provided is a method wherein the concentration of the therapeutic biological agent or salt thereof in the composition is from about 20 mg / mL to about 700 mg / mL.

[0008] In another aspect, the disclosure provides a method of treating an immune disorder in a subject in need thereof, comprising administering a pharmaceutically effective amount of: A plurality of particles suspended in a low viscosity pharmaceutically acceptable liquid carrier, each particle comprising at least one therapeutic biological agent or salt thereof. administering to a subject a composition comprising: the particles have less than about 10% aggregates of the therapeutic biological agent or salt thereof; and The method provides a method in which the concentration of the therapeutic biological agent or salt thereof in the composition is from about 20 mg / mL to about 700 mg / mL.

[0009] The disclosure herein provides a method of treating renal disease in a subject in need thereof, comprising administering a pharmaceutically effective amount of: A plurality of particles suspended in a low viscosity pharmaceutically acceptable liquid carrier, each particle comprising at least one therapeutic biological agent or salt thereof. administering to a subject a composition comprising: the particles have less than about 10% aggregates of the therapeutic biological agent or salt thereof; and Further provided is a method wherein the concentration of the therapeutic biological agent or salt thereof in the composition is from about 20 mg / mL to about 700 mg / mL.

[0010] In yet another aspect, the disclosure provides a method of treating a skin disease or condition in a subject in need thereof, comprising administering a pharmaceutically effective amount of: A plurality of particles suspended in a low viscosity pharmaceutically acceptable liquid carrier, each particle comprising at least one therapeutic biological agent or salt thereof. administering to a subject a composition comprising: the particles have less than about 10% aggregates of the therapeutic biological agent or salt thereof; and The method provides a method in which the concentration of the therapeutic biological agent or salt thereof in the composition is from about 20 mg / mL to about 700 mg / mL.

[0011] In yet another aspect, the present disclosure provides a pharmaceutically effective composition comprising: A plurality of particles suspended in a low viscosity pharmaceutically acceptable liquid carrier, each particle comprising at least one therapeutic biological agent or salt thereof. Includes; the particles have less than about 10% aggregates of the therapeutic biological agent or salt thereof; and The concentration of the therapeutic biological agent or its salt in the composition is about 20 mg / mL to about 700 mg / mL. / mL.

[0012] The disclosure herein provides a method of treating human immunodeficiency virus (HIV) infection in a subject in need thereof, comprising administering a pharmaceutically effective amount of: A plurality of particles suspended in a low viscosity pharmaceutically acceptable liquid carrier, each particle comprising at least one therapeutic biological agent or salt thereof. administering to a subject a composition comprising: the particles have less than about 10% aggregates of the therapeutic biological agent or salt thereof; and Also provided is a method wherein the concentration of the therapeutic biological agent or salt thereof in the composition is from about 20 mg / mL to about 700 mg / mL.

[0013] The compositions and methods may be useful for treating a disease or condition in a subject in need thereof, comprising administering a pharmaceutically effective amount of the composition to the subject. In a preferred embodiment, the treatment method employs a composition comprising a plurality of particles suspended in a low-viscosity pharmaceutically acceptable liquid carrier, each particle comprising at least one therapeutic biological agent or salt thereof; the particles having less than about 10% aggregates of the therapeutic biological agent or salt thereof; and the concentration of the therapeutic biological agent or salt thereof in the composition is from about 20 mg / mL to about 700 mg / mL. [Brief explanation of the drawings]

[0014] [Figure 1] 1 shows a plot of syringe force versus solids concentration using a 27 gauge ultra-thin wall (UTW) needle with a 4.7 mm internal diameter syringe barrel. [Figure 2A] 1 shows a plot of normalized ADCC % as a function of mAb concentration for FDA-labeled formulations and particle compositions at day 0. [Figure 2B] 1 shows a plot of normalized ADCC % as a function of mAb concentration for FDA-labeled formulations and particle compositions after 30 days of storage at 40° C. [Figure 3]1 shows graphs of the pharmacokinetic profiles for IV rituximab aqueous, SC rituximab aqueous, and SC rituximab particle (Elektroject) composition cohorts in rats (Sprague-Dawley). [Figure 4] Photographs of fluorescent images of SKH1 mice 1 minute, 2 days, and 7 days after receiving a SC injection of an aqueous formulation or particle composition (Elektroject) containing rituximab are shown. [Figure 5] 1 shows plots representing mean tumor volume growth curves for no treatment, SC rituximab aqueous, and SC rituximab composition (Elektroject) cohorts of mice (Fox Chase SCID). [Figure 6] 1 shows a graph of the concentration of mAb in the plasma of Yucatan minipigs normalized to body weight as a function of time for aqueous formulations and particulate compositions (Elektroject) after SC injection. [Figure 7] 1 shows a graph depicting viscosity and syringe force versus solid addition for an aqueous formulation and a particulate composition (Elektroject). DETAILED DESCRIPTION OF THE INVENTION

[0015] Therapeutic biological agents, particularly monoclonal antibody (mAb) therapeutics, have dramatically improved the treatment of human diseases. However, the delivery of these biological agents has placed a heavy burden on patients. The standard of administration is often by intravenous (IV) infusion at low concentrations, which can take several hours to deliver, cause discomfort to the patient, and increase the patient's risk of infection. Subcutaneous (SC) delivery by simple injection is preferred, but constraints on SC delivery volumes (1.5-2.0 mL) require antibody concentrations greater than 100 mg / mL, which is often impractical. Solution concentrations greater than 100 mg / mL are highly viscous, which results in very high injection forces and often enhances degradation of the antibody composition. Particle Suspension Technology The use of SC delivery allows for the delivery of therapeutic biological agents (e.g., antibodies) at concentrations >500 mg / mL while preserving the integrity of the therapeutic biological agent's (e.g., mAb) structure and bioactivity. Thus, shifting delivery of therapeutic biological agents from IV to SC can provide benefits to patients, healthcare providers, payers, and drug developers. In some cases, the concentration of biological agents in particulate form can result in less frequent SC injections for biological agents that are frequently injected SC.

[0016] The present disclosure generally relates to compositions and methods for treating (e.g., downregulating, reversing, inhibiting progression of, preventing) a disease or condition in a subject in need thereof, comprising administering to the subject a pharmaceutically effective amount of a composition comprising a plurality of particles suspended in a low-viscosity pharmaceutically acceptable liquid carrier, wherein each particle comprises at least one therapeutic biological agent or salt thereof; the particles have less than about 10% aggregates of the therapeutic biological agent or salt thereof; and the concentration of the therapeutic biological agent or salt thereof in the composition is from about 20 mg / mL to about 700 mg / mL. See Example 17 herein for a description of an example method for determining the concentration of a therapeutic biological agent in a composition of the present disclosure.

[0017] The present disclosure also relates to a method of treating cancer, an inflammatory disease or condition, an immune disease, a renal disease, a skin disease or condition, or human immunodeficiency virus (HIV) infection in a subject in need thereof, comprising administering to the subject a pharmaceutically effective amount of a composition comprising a plurality of particles suspended in a low-viscosity pharmaceutically acceptable liquid carrier, wherein each particle comprises at least one therapeutic biological agent or salt thereof; the particles have less than about 10% aggregates of the therapeutic biological agent or salt thereof; and the concentration of the therapeutic biological agent or salt thereof in the composition is from about 20 mg / mL to about 700 mg / mL. In certain embodiments, the compositions and methods described herein further comprise a pharmaceutically effective amount of at least one hyaluronan degrading agent, which can be administered simultaneously, sequentially, or intermittently with the composition. In a preferred embodiment, the hyaluronan degrading agent is hyaluronidase.

[0018] It is readily understood that the aspects and embodiments generally described herein are exemplary. The following more detailed description of various aspects and embodiments is not intended to limit the scope of the present disclosure, but is merely representative of various aspects and embodiments. Furthermore, the compositions and methods disclosed herein can be modified by those skilled in the art without departing from the scope of the present disclosure. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this disclosure belongs. All publications and patents mentioned herein are incorporated by reference.

[0019] Therapeutic Biological Agents Unless otherwise defined, all technical terms, notations, and other scientific terminology used herein are intended to have the meaning commonly understood by one of ordinary skill in the art to which this disclosure pertains. In some cases, terms with commonly understood meanings are defined herein for clarity and / or ready reference, and the inclusion of such definitions herein should not necessarily be construed as representing a substantial deviation from what is commonly understood in the art. The techniques and procedures described or referenced herein are generally well understood and commonly used by those skilled in the art using conventional methodology. Where appropriate, procedures involving the use of commercially available kits and reagents are generally performed according to manufacturer-specified protocols and / or parameters, unless otherwise noted. As used herein, the term "and / or," when used in a list of two or more items, means that any one of the listed items can be used alone, or any combination of two or more of the listed items can be used. For example, if a composition is described as including or excluding components A, B, and / or C, the composition may include A alone; B alone; C alone; a combination of A and B; a combination of A and C; a combination of B and C; or a combination of A, B, and C. The combination may be included or excluded.

[0020] A therapeutic biological agent, also known as a biological medical product or biopharmaceutical, is any pharmaceutical product manufactured in, extracted from, or semi-synthesized from a biological source. Therapeutic biological agents can include a wide range of products, such as vaccines, blood and blood components, allergens, somatic cells, gene therapy, tissues, and recombinant therapeutic proteins. In some embodiments, a therapeutic biological agent can be composed of sugars, proteins, or nucleic acids, or complex combinations of these substances, or can be a living entity, such as cells and tissues. Biological agents can be isolated from a variety of natural sources, such as humans, animals, or microorganisms, and can be produced by biotechnology methods or other techniques known in the art. Gene-based and cellular biological agents are often used, for example, to treat various medical conditions for which no other treatments are available. In certain embodiments of the present disclosure, the therapeutic biological agent is an antibody. In a preferred embodiment, the therapeutic biological agent is a monoclonal antibody (mAb).

[0021] The terms "antibody" and "immunoglobulin" are used interchangeably in the broadest sense and include, but are not limited to, monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, multispecific antibodies, e.g., bispecific antibodies, humanized antibodies, fully human antibodies, biparatopic antibodies, humanized camelid heavy chain antibodies, and non-human / human chimeric antibodies, regardless of how they are produced, i.e., using immunization, recombinant, or synthetic methodologies. A "parent antibody" is an antibody obtained by exposure of the immune system to an antigen prior to modification of the antibody for its intended use, e.g., humanization of a non-human antibody for use as a human therapeutic antibody. Antibodies may be gamma globulin proteins found in the blood or other body fluids of vertebrates, which function in the immune system to bind antigens and thus identify and / or neutralize foreign substances. Antibodies can be assigned to different classes or isotypes. There are five classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, each with heavy chains designated alpha, delta, epsilon, gamma, and mu, respectively. The gamma class is further divided into subclasses based on differences in sequence and function, for example, humans express the following subclasses: IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. Light chains from any vertebrate species can be assigned to one of two distinct types, e.g., kappa and lambda, based on the amino acid sequence of their constant domain.

[0022] 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 kappa or lambda. In other embodiments, heavy chains are classified as gamma, mu, alpha, delta, or epsilon, which define the immunoglobulin classes IgG, IgM, IgA, IgD, and IgE, respectively. In other embodiments of the disclosure, the antibody is a human antibody. In certain embodiments, the human antibody is an IgG antibody. In certain other embodiments, the IgG antibody is an IgG1 antibody. In preferred embodiments of the disclosure, the antibody is a monoclonal antibody.

[0023] An exemplary antibody (immunoglobulin) structural unit comprises a tetramer. Each tetramer is composed of two identical pairs of polypeptide chains, each pair having one "light" chain (about 25 kD) and one "heavy" chain (about 50-70 kD). The N-terminus of each chain defines a variable region of about 100-110 or more amino acids primarily responsible for antigen recognition. The terms "variable light" chain, domain, region, and component are used interchangeably and are referred to as "VL" or "V L abbreviated as " , refers to the light chain of an antibody or antibody fragment. Similarly, the terms "variable heavy" chain, domain, region, and component are used interchangeably and may be referred to as "VH" or "V H " and antibody or antibody The term "antibody" refers to the heavy chain of a fragment of an antibody. Antibodies are generally heterotetrameric glycoproteins composed of two identical light (L) chains and two identical heavy (H) chains. Each L chain contains one covalently linked Each H chain is linked to an H chain by a synthetic disulfide bond. Two H chains are linked to each other by one or more disulfide bonds depending on the H chain isotype. Each H chain and L chain also has regularly spaced intrachain disulfide bridges. The H chain and L chain define a specific Ig domain. Specifically, each H chain has a variable domain (VH) at the N-terminus, followed by three constant domains (CH) for each of the alpha and gamma chains and four CH domains for the p and c isotypes. Each L chain has a variable domain (VL) at the N-terminus, followed by a constant domain (CL) at the 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 domains comprise the Fc portion, which includes 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, the portion recognized by Fc receptors (FcRs) that are also found on certain types of cells.

[0024] As disclosed herein, the pairing of a VH and a VL together forms a "variable region" or "variable domain" comprising the amino-terminal domain of either the heavy or light chain of an antibody. The variable domain of a heavy chain may be referred to as "VH." The variable domain of a light chain may be referred to as "VL." The variable domain contains an "antigen-binding site" that influences antigen binding and defines the specificity of a particular antibody for its particular antigen. The variable region spans approximately 110 amino acid residues and consists of relatively invariant stretches of 15-30 amino acids called framework regions (FRs) (generally about four) separated by shorter, highly variable regions called "hypervariable regions" (generally about three), each generally 9-12 amino acids long. The FRs largely adopt a p-sheet configuration, and the hypervariable regions form loops that connect and occasionally form part of the p-sheet structure. In certain embodiments, "hypervariable region" refers to the region of an antibody variable domain that is hypervariable in sequence and / or forms structurally defined loops. Generally, antibodies contain six hypervariable regions: three in the VH (H1, H2, H3) and three in the VL (L1, L2, L3). "Framework" or "FR" residues are those variable domain residues other than the hypervariable region residues as herein defined.

[0025] The terms "full length antibody," "intact antibody," and "whole antibody" are used interchangeably herein to refer to an antibody in its substantially intact form, and not as an antibody fragment as defined above. These terms particularly refer to an antibody having a heavy chain containing an Fc region. A full length antibody may be a native sequence antibody or an antibody variant. In certain embodiments, an "intact" or "whole" antibody is one that comprises an antigen-binding site and a CL and at least heavy chain constant domains, CH1, 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.

[0026] As indicated above, the term antibody, as used herein, includes fragments of antibodies that retain the ability to specifically interact with, e.g., bind to, an antigen, unless otherwise stated or clearly contradicted by context. It has been shown that the antigen-binding function of an antibody can be performed by fragments of a full-length antibody. Examples of binding fragments encompassed within the term "antibody" include: (i) Fab' or Fab fragments, monovalent fragments consisting of a light chain variable domain (VL), a heavy chain variable domain (VH), a light chain constant region (CL), and a heavy chain constant region domain 1 (CH1) domain, or monovalent antibodies as described in WO 2007 / 059782; (ii) F(ab')2 fragments (iii) a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; (iii) an Fd fragment consisting essentially of a VH and a CH1 domain; (iv) an Fv fragment consisting essentially of the VL and VH domains of a single arm of an antibody; (v) an Fv fragment consisting essentially of a VH domain, also called a domain antibody (Hol (t et al; Trends Biotechnol. 2003 November; 21(11):484-90) dAb fragments (Ward et al., Nature 341, 544-546(1989)); (vi) camelids or nanobodies (Revets et al; Expert Opin Biol Ther. 2005 January; 5(1):111-24), and (vii) isolated complementarity determining regions (CDRs). Furthermore, although the two domains of an Fv fragment, VL and VH, are encoded by separate genes, they can be joined using recombinant methods by a synthetic linker that allows them to be produced as a single protein chain in which the VL and VH regions pair to form a monovalent molecule (known as a single-chain antibody or single-chain Fv (scFv), see, e.g., Revets et al.; Expert Opin Biol Ther. 2005 January; 5(1):111-24 and Bird et al., Science 242, 423-426 (1988)). Such single-chain antibodies are encompassed within the term antibody unless otherwise specified or clearly indicated by context. While such fragments are generally included within the meaning of antibody, they collectively and individually are distinctive features of the present invention and exhibit distinct biological properties and utilities. These and other useful antibody fragments are discussed further herein in connection with the present invention.

[0027] As disclosed herein, "whole antibody fragments comprising the variable domain" include Fab, Fab', F(ab')2 and Fv fragments; diabodies; linear antibodies; single chain antibodies; These include antibody molecules; and multispecific antibodies formed from antibody fragments. A "Fab fragment" consists of an entire L chain together with the variable region domain of the H chain and the first constant domain (CHI) of one heavy chain. Each Fab fragment is monovalent with respect to antigen binding, i.e., it has a single antigen-binding site. A "Fab' fragment" differs from a Fab fragment by having a few additional 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 ​​residues of the constant domains bear a free thiol group. A "F(ab')2 fragment" has bivalent antigen-binding activity. It roughly corresponds to two disulfide-linked Fab fragments with the same affinity and is still capable of cross-linking antigen. "Fv" is the minimum antibody fragment containing 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 single-chain Fv (scFv) species, one heavy- and one light-chain variable domain can be covalently linked by a flexible peptide linker so that the light and heavy chains can associate in a "dimeric" structure similar to that in two-chain Fv species. The folding of these two domains results in six hypervariable loops (three loops from each H and L chain) that provide amino acid residues for antigen binding and confer antigen-binding specificity to the antibody. "Single-chain Fv," also abbreviated as "sFv" or "scFv," is an antibody fragment containing VH and VL antibody domains connected to form a single-chain polypeptide. In preferred 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 the antigen) that has the ability to recognize and bind antigen, although at a lower affinity than the entire binding site.

[0028] In some embodiments, "diabody" refers to an antibody fragment with two antigen-binding 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). Small antibody fragments comprise sFv fragments with a short linker (about 5-10 residues) between the VH and VL domains such that interchain pairing but not intrachain pairing of the variable domains is achieved. Diabodies are prepared by constructing fragments of sFv fragments, resulting in bivalent fragments, i.e., fragments with two antigen-binding sites. In other embodiments, diabodies can 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. Trispecific and tetraspecific antibodies are also generally known in the art.

[0029] An "antibody fragment" or "antigen-binding fragment" of an antibody as described herein includes only a portion of the intact antibody that generally comprises the antigen-binding site of the intact antibody and thus retains the ability to bind antigen. Illustrative examples of antibody fragments encompassed by the present disclosure include, but are not limited to, (i) a Fab fragment having the VL, CL, VH, and CH1 domains; (ii) a Fab' fragment, which is a Fab fragment with one or more cysteine ​​residues at the C-terminus of the CH1 domain; (iii) an Fd fragment having the VH and CH1 domains; (iv) an Fd' fragment having the VH and CH1 domains and one or more cysteine ​​residues at the C-terminus of the CH1 domain; (v) an Fv fragment having the VL and VH domains of a single arm of the antibody; (vi) a dAb fragment consisting of the VH domain; (vii) isolated CDR regions; and (viii) a F(ab')2 fragment (resp. a fragment separated by disulfide bridges in the hinge region). (ix) single-chain antibody molecules (e.g., single-chain Fv; scFv); (x) "diabodies" having two antigen-binding sites comprising a heavy-chain variable domain (VH) connected to a light-chain variable domain (VL) in the same polypeptide chain; and (xi) "linear antibodies" comprising a pair of tandem Fd segments (VH-CH1-VH-CH1) that, 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 comprising at least the hypervariable and framework regions required to confer antigen-binding function to the V domain. An antigen-binding site can be used in the methods described herein to bind to an antibody or antibody fragment (e.g., dAb, Fab, Fd, Fv, F(ab')2, or scFv). ) In certain embodiments, the therapeutic biological agent is an antibody fragment.

[0030] In some embodiments, the term "single-chain Fv" or "scFv" or "single-chain" antibody can refer to an antibody fragment comprising the VH and VL domains of an 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 that enables the sFv to form the desired structure for antigen binding. For a review of sFvs, see Pluckthun, THE PHARMACOLOGY OF MONOCLONAL ANTIBODIES, vol. 113, Rosenburg and Moore eds. Springer-Verlag, New York, pp. 269-315 (1994).

[0031] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible natural mutations that may be present in minor amounts. Monoclonal antibodies (mAbs) are highly specific, targeting a single antigenic site or determinant on an antigen. In addition to their specificity, monoclonal antibodies are advantageous in that they may be synthesized uncontaminated by other antibodies. Monoclonal antibodies may be prepared by hybridoma methodology. Monoclonal antibodies may also be isolated from phage antibody libraries using molecular engineering techniques. The monoclonal antibodies of the present disclosure may be generated by recombinant DNA methodology and may be referred to as "recombinant antibodies" or "recombinant monoclonal antibodies" as disclosed herein. In some embodiments, a monoclonal antibody is a single species of antibody, in which all antibody-producing cells are derived from a single B-lymphocyte cell lineage and therefore all antibody molecules target the same epitope. We recognize that methods for raising 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 raising monoclonal antibodies. In certain embodiments, rabbit, sheep, or frog cells are used in raising monoclonal antibodies. The use of rats is known and may offer certain advantages. Mice, such as BALB / c mice, are routinely used and generally yield a high percentage of stable fusions. In yet other embodiments of the present disclosure, the antibody is a monoclonal antibody. In certain embodiments of the present disclosure, the antibody is a human monoclonal antibody. In a preferred embodiment, the IgG antibody is a human monoclonal antibody.

[0032] In other embodiments, recombinant antibody fragments can be isolated from phage antibody libraries using techniques known in the art. See, e.g., Clackson et al., 1991, Nature 352:624-628; Marks et al., 1991, J. Mol. Biol. 222:581-597. Recombinant antibody fragments can 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) can be used to generate recombinant full-length immunoglobulins using methods known in the art (see, e.g., Persic et al., 1997, Gene 187:9-18).

[0033] An "isolated antibody" is one that has been identified and separated and / or recovered from components of its existing environment. Contaminant components are materials that would interfere with therapeutic uses for the antibody, and may include enzymes, hormones, and other proteinaceous or non-proteinaceous solutes.

[0034] As used herein, "human antibody" refers to an antibody having an amino acid sequence corresponding 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 antigen to transgenic animals that have been engineered to produce such antibodies in response to antigen exposure, but whose endogenous gene loci are impaired. "Humanized" forms of non-human, e.g., rodent, antibodies are chimeric antibodies that contain minimal sequence derived from the non-human antibody. As used herein, a "chimeric antibody" is an antibody having variable domains from a first antibody and constant domains from a second antibody, where (i) the first and second antibodies are from different species (Morrison et al., (1984) Proc. Natl. Acad. Sci. USA 81:6851-6855) or (ii) the first and second antibodies are from different isotypes, e.g., variable domains from an IgG1 antibody and constant domains from an IgG4 antibody. In some embodiments, the variable domain is derived from a non-human antibody, such as a murine antibody (the "parent antibody"), and the constant domain sequences are derived from a human antibody. In other embodiments, the variable domain is a humanized variable domain from a murine antibody and a human antibody constant domain. 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 performance. In some cases, 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 the donor antibody. These modifications are made to further refine antibody performance. Generally, humanized antibodies 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 the non-human immunoglobulin, and the F All or substantially all of R are FRs of a human immunoglobulin sequence. The humanized antibody optionally also comprises at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin.

[0035] An "affinity matured" antibody is one that has one or more alterations in one or more hypervariable regions thereof that result in an improvement in the affinity of the antibody for an antigen, compared to a parent antibody that does not possess those alterations. In some embodiments, an affinity matured antibody may have micromolar affinity for the target antigen. In other embodiments, an affinity matured antibody may have nanomolar or even picomolar affinity for the target antigen. Affinity matured antibodies are produced by procedures known in the art.

[0036] A "blocking" or "antagonist" antibody is one that inhibits or reduces the biological activity of the antigen to which it binds. In some embodiments, a blocking or antagonist antibody substantially or completely inhibits the biological activity of the antigen. An "agonist antibody," as used herein, is an antibody that mimics at least one functional activity of a polypeptide of interest. As used herein, the terms "inhibition," "inhibit," "inhibiting," and the like, with respect to a protein inhibitor, e.g., an antagonist, interaction, mean to adversely affect, e.g., reduce, the activity or function of a protein relative to the activity or function of the protein in the absence of the inhibitor. In some embodiments, inhibition refers to a reduction in a disease or disease symptom. In other embodiments, inhibition refers to a reduction in the activity of a signal transduction pathway or signaling pathway. Thus, inhibition includes at least partially or completely blocking a stimulus; reducing, preventing, or delaying activation; or inactivating, desensitizing, or downregulating signal transduction, enzyme activity, or the amount of a protein.

[0037] "Binding affinity" generally refers to the strength of the sum of non-covalent interactions between a single binding site of a molecule, e.g., an antibody, and its binding partner, e.g., an antigen. Unless otherwise indicated, as used herein, "binding affinity" refers to the intrinsic binding affinity, reflecting a 1:1 interaction between members of a binding pair, e.g., an antibody and an antigen. The affinity of 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 easily, while high-affinity antibodies generally bind antigen more quickly and tend to remain bound longer. Various methods for 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 portion of an antigen bound by the antigen-binding site of an antibody. In some embodiments, an epitope can 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 found in the primary protein structure. In another embodiment, the epitope is a "conformational epitope," ie, a hypervariable loop of a CDR binds to residues that are displayed in the tertiary or quaternary structure of a protein.

[0038] In some embodiments of the compositions and methods described above, the therapeutic biological agent is an antibody. In other embodiments, the antibody is selected from the group consisting of, but not limited to, 3F8, abagovomab, abatacept, abciximab, abiutuzumab, abrezekimab, abrilumab, acritumomab, actoxumab, abituzumab, adalimumab-adbm, adalimumab-atto, adalimumab-bwwb, adecatumumab, Ado-trastuzumab emtansine, and adu Kanumab, afasevicumab, afelimomab, aflibercept, afutuzumab, aracizumab pegol, ALD518, alefacept, alemtuzumab, alirocumab, altumomab pentetate, amatuximab, anatumomab mafenatox, andecaliximab, anetumab ravtansine, anifrolumab, anrukinzumab, apolizumab, Purtumab ixadotin, arcitumomab, asclinicalvacuumab, acelizumab, atezolizumab, atidortoxumab, atinumab, atlizumab, atolimu-mab, avelumab, azintuximab vedotin, bapineuzumab, basiliximab, bavituximab, BCD-100, brosozumab, begelomab, belantamab mafodotin, belatacept, belimumab, bemarituzumab, benralizumab, bermekimab, bersanlimab, bertilimumab, besilesomab, bevacizumab, bevacizumab-awwb, bezlotoxumab, biciromab, bimaglu Mab, bimekizumab, viltamimab, bivatuzumab mertansine, bleselumab, blinatumomab, brontuzumab, brosozumab, bococizumab, brazikumab, brentuximab vedotin, briakinumab, brodalumab, brolucizumab, brontiximab, burosumab, cabilalizumab, camidalumab tesirin, camrelizumab, canakinumab, cantuzumab mertansine, cantuzumab ravtansine, caplacizumab, capromab pendetide, carlumab, carotuximab, catumaxomab, cBR96-doxorubicin immunoconjugate, sede Lizumab, cemiplimab, sergituzumab amnaleukin, sergituzumab amnaleukin, certolizumab pegol, cetrelimab, cetuximab, civisatamab, cirmtuzumab, sitatuzumab bogatox, cixutumumab, clazakizumab, clenoliximab, clivatuzumab tetraxetan, codrituzumab, cofetuzumab peridotin, coltuximab ravtansine, conatumumab, concizumab, cosfrobiximab, crenezumab, CR6261, crizanlizumab, clotezumab, cusatuzumab, dacetuzumab, daclizumab, darotuzumab , dapirolizumab pegol, daratumumab, dectrecumab, demcizumab, denileukin diftitox, denintuzumab mafodotin, denosumab, depatuxizumab mafodotin, dellotuximab biotin, detumomab, dezamizumab, dinutuximab, zilidabumab, domagrozumab, dorlimomab alitox, dostarlimab, drozitumab, DS-8201, zurigotumab, dupilumab, zulvalumab, zusigitumab, zuvortuxizumab, ecromeximab, eculizumab, edovacomab, edrecolomab, efalizumab, efungumab,Eldelumab, elezanumab, elgemtumab, elotuzumab, ersilimomab, emactuzumab, emibetuzumab, emicizumab, enapotamab vedotin, enavatuzumab, enfortumab vedotin, enlimomab pegol, enoblitzumab, enokizumab, enoticumab, ensituximab, epitumomab cituxetan, epoetin-alfa, epoetin-alfa-epbx, epratuzumab, eptinezumab, erenumab, erlizumab, ertumaxomab, etanercept, etanercept-szzs, etaracizumab, etigilimab, etoro Lisumab, evinacumab, evolocumab, exibirumab, Factor VIII Fc fusion protein, Factor IX Fc fusion protein, fanolesomab, faralimomab, faricimab, farletuzumab, fasinumab, felvizumab, fezakinumab, fivatuzumab, ficlatuzumab, figitumumab, filgrastim, filgrastim-sndz, filibumab, framvotumab, fretikumab, flotetuzumab, hontolizumab, foramumab, foravilumab, fremanezumab, fresolimumab, flobocimab, furunvetomab, furanumab , futuximab, galcanezumab, galiximab, ganitumab, gantenerumab, gatipotuzumab, gavilimomab, gedivumab, gemtuzumab ozogamicin, gevokizumab, gilvetomab, gimcilumab, girentuximab, glenbatumumab vedotin, golimumab, gomiliximab, goslanemab, guselkumab, ibalizumab, IBI308, ibritumomab tiuxetan, icrucumab, idarucizumab, ifavotuzumab, igovomab, iradatuzumab vedotin, IMAB362, imalumab, imaprelimab, imciromab, imgatuzumab , inlacumab, indatuximab ravtansine, indusatumab vedotin, inebilizumab, infliximab, infliximab-abda, infliximab-dyyb, infliximab-qbtx, intetumumab, inolimomab, inotuzumab ozogamicin, ipilimumab, iomab-B, iratumumab, isatuximab, iscalimab, istiratumab, itolizumab, ixekizumab, keliximab, labetuzumab, lacunotuzumab, radatuzumab vedotin, lambrolizumab, lampalizumab, lanadelumab, landrozumab, la,Prituximab emtansine, ralcabiximab, lebrikizumab, remaresomab, lendalizumab, lembervimab, lenzilumab, lerdelimumab, leronlimab, lesofabumab, letolizumab, lexatumumab, ribivirumab, rifastuzumab vedotin, ligelizumab, loncastuximab tesirin, rosatuximab vedotin, rilotumab satetraxetan, lintuzumab, lirilumab, roderucizumab, lokivetmab, lorvotuzumab mertansine, lucatumumab, lurizumab pegol, rumiliximab, lumletuzumab, rupartumab amd Chin, rutikizumab, mapatumumab, margetuximab, marstacimab, maslimomab, mavrilimumab, matuzumab, mepolizumab, metelimumab, milatuzumab, minletumomab, mirikizumab, mirvetuximab soravtansine, mitumomab, modotuximab, mogamulizumab, monalizumab, morolimumab, mosunetuzumab, motavizumab, moxetumomab pasudotoxin, muromonab-CD3, nacolomab butafenatox, namilumab, naptumomab estafenatox, naratuximab emtansine, narutomab, natalizumab, nabiscosides Xixizumab, navivumab, naxitamab, nebacumab, necitumumab, nemolizumab, NEOD001, nerelimomab, nesbacumab, netakimab, nimotuzumab, nirsevimab, nivolumab, nofetumomab merpentan, obilutoxaximab, obinutuzumab, ocralatuzumab, ocrelizumab, ozlimomab, ofatumumab, olaratumab, oleculab, orendalizumab, olokizumab, omalizumab, omburtamab, OMS721, onartuzumab, ontuxizumab, onvatilimab, opicinumab, oportuzumab monatox, Legovomab, olticumab, otelixizumab, otilimab, otlertuzumab, oxelumab, ozanezumab, ozoralizumab, pagibaximab, palivizumab, pamrevlumab, panitumumab, pancomab, panobacumab, palsatuzumab, pascolizumab, pasotuxizumab, pateclizumab, patritumab, PDR001, pegfilgrastim-jmdb, pembrolizumab, pemtumomab, perakizumab, pertuzumab, pexelizumab, pidilizumab, pinatuzumab vedotin, pintumomab, placurab, prosalizumab, pogalizumab,Polatuzumab vedotin, ponezumab, polgabiximab, prasinezumab, prezalizumab, priliximab, plitoxaximab, pritumumab, PRO140, kilimumab, racotumomab, radletumab, rafivirumab, ralpancizumab, ramucirumab, ranevetomab, ranibizumab, raxibacumab, ravagalimab, ravuturizumab, refanezumab, regavirumab, relatolimab, lemtolumab, reslizumab, rilonacept, rilotumumab, rinukumab, risankizumab, rituximab, rituximab-abbs, rituximab-pvvr ... Babazumab pegol, rivabazumab pegol, lobatumumab, Rmab, lorezumab, romilukimab, romiplostim, romosozumab, rontalizumab, rosmantuzumab, rovalpituzumab tesirin, rovalpituzumab tesirin, rovelizumab, rozanolixizumab, ruplizumab, sacituzumab govitecan, samalizumab, samuraizumab vedotin, sapelizumab, sarilumab, satralizumab (SA237), satumomab pendetide, secukinumab, selicrelumab, seribantumab, cetoxaximab, ceturusumab, sevilumab, sibrotuzumab, SG N-CD19A, SGN-CD33A, SHP647, sifalimumab, siltuximab, simtuzumab, siplizumab, siltratumab vedotin, sirukumab, sofituzumab vedotin, solanezumab, solitomab, sonepcizumab, sontuzumab, spartalizumab, stamulumab, sulesomab, sputumumab, stimulimab, subizumab, sublatoxumab, tabalumab, tacatuzumab tetraxetan, tadocizumab, talaxuzumab, talizumab, tamtubetumab, tanezumab, tapritumomab paptox, tarexuzumab, tabolimab, tefiba Zumab, terimomab alitox, telisotuzumab vedotin, tenatumomab, teneliximab, teplizumab, tepositamab, teprotumumab, tesidolumab, tetulomab, tezepelumab, TGN1412, tiburizumab, ticilimumab, tildrakizumab, tigatuzumab, timigituzumab, timolumab, tiragotumab, tislelizumab, tisotuzumab vedotin, TNX-650, tocilizumab, tomzotuximab, toralizumab, tosatoxumab, tositumomab, tobetumab, tralokinumab, trastuzumab, trastuzumab-anns, trastuzumab-, dkst, trastuzumab-dttb, trastuzumab emtansine, trastuzumab-pkrb, tregalizumab, tremelimumab, trevoglumab, tucotuzumab-celmoleukin, tuvilumab, ublituximab, urocupulumab, urelumab, urtoxazumab, ustekinumab, utomilumab, vadastuximab-butarilin, banalimab, bundlestuzumab vedotin, vanticutuzumab, vanucizumab, bapaliximab, valisacumab, varlilumab , batelizumab, vedolizumab, veltuzumab, bepalimomab, besencumab, visilizumab, bovalilizumab, volociximab, bonlerolizumab, bopratelimab, borsetuzumab mafodotin, votumumab, xentuzumab, XMAB-5574, zalutumumab, zanolimumab, zatuximab, xenoctuzumab, ziralimumab, zolbetuximab (IMAB362, claudiximab), Ziv-aflibercept, or zolimomab alitoxin.

[0039] In other embodiments of the above compositions and methods, the antibody is monoclonal. In certain embodiments, the monoclonal antibody is selected from the group consisting of, but not limited to, 3F8, 8H9, abatacept, abagovomab, abciximab, abituzumab, adalimumab-adbm, adalimumab-atto, adalimumab-bwwb, abrelumab, actoxumab, abituzumab, abrezekimab, abrelumab, actoxumab, adalimumab, adecatumumab, adotrastuzumab emtansine, aducanumab, afacevicumab, afelimomab, afribe Rucept, afutuzumab, aracizumab pegol, ALD518, alefacept, alemtuzumab, alirocumab, altumomab pentetate, amatuximab, anatumomab mafenatox, andecaliximab, anetumab ravtansine, anifrolumab, anrukinzumab (IMA-638), apolizumab, arcitumomab, asclinbacumab, acelizumab, atezolizumab, atidortoxumab, atinumab, atlizumab (tocilizumab), atorlim Mab, avelumab, bapineuzumab, basiliximab, bevacizumab, bevacizumab-awwb, BCD-100, brosozumab, begelomab, belatacept, belimumab, bemarituzumab, benralizumab, bermekimab, bersanlimab, bertilimumab, besilesomab, bezlotoxumab, biciromab, bimagrumab, bimekizumab, vilutamimab, bivatuzumab mertansine, bleselumab, blinatumomab, brontuzumab, brosozumab, bococizumab , brazikumab, brentuximab vedotin, briakinumab, brodalumab, brolucizumab, brontixutuzumab, burosumab, cabilalizumab, camrelizumab, canakinumab, cantuzumab mertansine, cantuzumab ravtansine, caplacizumab, capromab pendetide, carlumab, carotuximab, catumaxomab, cedelizumab, cemiplimab, certolizumab pegol, cetrelimab, cetuximab, civisatamab, cimutuzumab, Ch.14.18, sitatuzumab bogatox, cixutumumab, clazakizumab, clenoliximab, clivatuzumab tetraxetan, codrituzumab, cofetuzumab peridotin, coltuximab ravtansine, conatumumab, concizumab, cosfrobiximab, crenezumab, CR6261, crizanlizumab, clotezumab, cusatuzumab, dacetuzumab, dac Lizumab, dalotuzumab, dapirolizumab pegol, daratumumab, dectrecumab, demcizumab, denileukin diftitox, denintuzumab mafodotin, denosumab, delrotuximab biotin, detumomab, dezamizumab, dinutuximab, zilidabumab, domagrozumab, dorlimomab alitox, dostarlimab, drozitumab, zurigotumab, Dupilumab, Zulvalumab, Zusigitumab, Zvortuxizumab, Eclomeximab, Eculizumab, Edobacomab, Edrecolomab, Efalizumab, Efungumab, Eldelumab, Elezanumab, Elgemtuzumab, Elotuzumab, Elcilimomab, Emactuzumab, Emibetuzumab, Emicizumab, Enavatuzumab, Enfortumab vedotin, Enrimo Mab pegol, enoblituzumab, enokizumab, enoticumab, encituximab, epitumomab cituxetan, epoetin-alfa, epoetin-alfa-epbx, epratuzumab, eptinezumab, erenumab, erlizumab, ertumaxomab, etanercept, etanercept-szzs, etaracizumab, etigilimab, etorolizumab, et. Binacumab, evolocumab, exibirumab, Factor VIII Fc fusion protein, Factor IX Fc fusion protein, fanolesomab, faralimomab, faricimab, farletuzumab, fasinumab, FBTA05, felvizumab, fezakinumab, fivatuzumab, ficlatuzumab, figitumumab, filgrastim, filgrastim-sndz, filibumab, framvotumab, fretikumab, flotetuzumab, hontolizumab, foramumab, foravilumab, fremanezumab, fresolimumab, flobocimab, furunvetomab, furanumab Mab, futuximab, galcanezumab, galiximab, ganitumab, gantenerumab, gatipotuzumab, gavilimomab, gezivumab, gemtuzumab ozogamicin, gevokizumab, gilvetomab, gimcilumab, girentuximab, glenbatumumab vedotin, golimumab, gomiliximab, goslanemab, guselkumab, ibalizumab, IBI308, ibritumomab tiuxetan, icrucumab, idarucizumab, ifavotuzumab, igovomab, IMAB362, imalumab, imaprelimab, imciromab, imgatuzumab, inlacumab, in Datuximab ravtansine, indusatumab vedotin, inebilizumab, infliximab, infliximab-abda, infliximab-dyyb, infliximab-qbtx, intetumumab, inolimomab, inotuzumab ozogamicin, ipilimumab, iratumumab, isatuximab, iscalimab, istiratumab, itolizumab, ixekizumab, keliximab, labetuzumab, lacunotuzumab, lambrolizumab, lampalizumab, lanadelumab, landgrozumab, ralcabiximab, lebrikizumab, lemaresomab, and lendarizumab , lembervimab, lenzilumab, lerdelimumab, leronlimab, lesofabumab, letolizumab, lexatumumab, ribivirumab, rifastuzumab vedotin, ligelizumab, rilotumab satetraxetan, lintuzumab, lirilumab, roderucizumab, lokivetmab, lorvotuzumab mertansine, lucatumumab, lurizumab pegol, lumiliximab, lumletuzumab, lutikizumab, mapatumumab, margetuximab, marstacimab, maslimomab, mavrilimumab, matuzumab, mepolizumab, metelimumab, milatuzumab, minletumomab,Mirikizumab, mirvetuximab soravtansine, mitumomab, modotuximab, mogamulizumab, monalizumab, morolimumab, mosunetuzumab, motavizumab, moxetumomab pasudotoxin, muromonab-CD3, nacolomab butafenatox, namilumab, naptumomab estafenatox, narunatumumab, natalizumab, nabicixizumab, naxitamab, nebacumab, necitumumab, nemolizumab, NEOD001, nerelimomab, nesbacumab, netakimab, nimotuzumab, nirsevimab, nivolumab, nofetumomab merpen Tan, obiltoxaximab, obinutuzumab, ocralizumab, ocrelizumab, ozlimomab, ofatumumab, olaratumab, oleculab, orendalizumab, olokizumab, omalizumab, omburtamab, OMS721, onartuzumab, ontuxizumab, onvatilimab, opicinumab, oportuzumab monatox, oregovomab, olticumab, otelixizumab, otilimab, otreltuzumab, oxelumab, ozanezumab, pagibaximab, palivizumab, pamrevlumab, panitumumab, pancomab, panobacumab, pulsat Ibuprofen, pascolizumab, pasotuximab, pateclizumab, patritumab, PDR001, pegfilgrastim-jmdb, pembrolizumab, pemtumomab, perakizumab, pertuzumab, pexelizumab, pidilizumab, pinatuzumab vedotin, pintumomab, placurumab, prosalizumab, pogalizumab, polatuzumab vedotin, ponezumab, polgabiximab, prasinezumab, prezalizumab, priliximab, pritoxaximab, pritumumab, PRO140, kilimumab, tetulomab, racotumomab, radletumab, rafivirumab , ralpancizumab, ramucirumab, ranevetomab, ranibizumab, raxibacumab, ravagalimab, ravutolizumab, refanezumab, regavirumab, relatolimab, lemtolumab, reslizumab, rilonacept, rilotumumab, linukumab, risankizumab-rzaa, rituximab, rituximab-abbs, rituximab-pvvr, lobatumumab, Rmab, lorezumab, romilkimab, romiplostim, romosozumab, rontalizumab, rosmantuzumab, rovelizumab, rozanolixizumab, ruplizumab, sacituzumab govitecan, sumari,Ibuprofen, sarilumab, satralizumab (SA237), satumomab pendetide, secukinumab, cericlerumab, seribantumab, cetoxaximab, setorsumab, sevirumab, sibrotuzumab, SGN-CD19A, SGN-CD33A, SHP647, sifalimumab, siltuximab, simtuzumab, siplizumab, sirukumab, sofituzumab vedotin, solanezumab, solitomab, sonepcizumab, sontuzumab, spartalizumab, stamulumab, sulesomab, sputavumab, stimulimab, suvizumab, subratoxumab , tabalumab, tacatuzumab tetraxetan, tadocizumab, talaxuzumab, talizumab, tamtubetumab, tanezumab, taplitumomab paptokis, tarexuzumab, tabolimab, tefibazumab, terimomab alitokis, tenatumomab, teneliximab, teplizumab, tepositamab, teprotumumab, tesidolumab, tetulomab (rilotomab), tezepelumab, TGN1412, tiburizumab, ticilimumab (tremelimumab), tildrakizumab, tigatuzumab, timigtuzumab, timolumab, tiragotumab, tislelizumab, TNX-650, tocilizumab (atlizumab), tomzotuximab, toralizumab, tosatoxumab, tositumomab, tobetumab, tralokinumab, trastuzumab, trastuzumab-anns, trastuzumab-dkst, trastuzumab-dttb, trastuzumab emtansine, trastuzumab-pkrb, TRBS07, tregalizumab, tremelimumab, trevoglumab, tucotuzumab-celmoleukin, tuvilumab, ublituximab, urocuplumab, urelumab, urtoxazumab, ustekinumab, utomilumab, Limab, bundletuzumab vedotin, vanticutumab, vanucizumab, bapaliximab, valisacumab, varlilumab, batelizumab, vedolizumab, veltuzumab, bepalimomab, besencumab, visilizumab, bovalilizumab, volociximab, bonlerolizumab, bopratelimab, borsetuzumab mafodotin, votumumab, xentuzumab, XMAB-5574, zalutumumab, zanolimumab, zatuximab, xenoctuzumab, ziralimumab, zolbetuximab (IMAB362, claudiximab), Ziv-aflibercept,The monoclonal antibody comprises rituximab, rituximab-abbs, or rituximab-pvvr.

[0040] In some embodiments, the monoclonal antibody is a biosimilar. In other embodiments, biosimilars include, but are not limited to, adalimumab-adbm, adalimumab-atto, adalimumab-bwwb, bevacizumab-awwb, epoetin alfa-epbx, etanercept-szzs, infliximab-abda, infliximab-dyyb, infliximab-qbtx, filgrastim-sndz, pegfilgrastim-jmdb, pegfilgrastim-bmez, risankizumab-rzaa, rituximab-abbs, rituximab-pvvr, trastuzumab-anns, trastuzumab-dttb, trastuzumab-pkrb, or trastuzumab-dkst. In certain embodiments, the active biosimilar substance is adalimumab, bevacizumab, enoxaparin sodium, epoetin alfa, epoetin zeta, etanercept, filgrastim, follitropin alfa, infliximab, insulin glargine, insulin lispro, pegfilgrastim, risankizumab, rituximab, rituximab-abbs, rituximab-pvvr, somatropin, teriparatide, or trastuzumab. In preferred embodiments, the biosimilar is rituximab, rituximab-abbs, or rituximab-pvvr.

[0041] 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) variant, a multispecific antibody, a bispecific antibody, a chimeric antibody, a humanized antibody, a human antibody, a fusion protein containing an antigenic determinant portion of an antibody, or other modified immunoglobulin molecule containing an antigen recognition site.

[0042] Provided herein are compositions and methods for treating cancer in a subject in need thereof, comprising administering to the subject a pharmaceutically effective amount of a composition comprising a plurality of particles suspended in a low-viscosity pharmaceutically acceptable liquid carrier, wherein each particle comprises at least one therapeutic biological agent or salt thereof; the particles have less than about 10% aggregates of the therapeutic biological agent or salt thereof; and the concentration of the therapeutic biological agent or salt thereof in the composition is from about 20 mg / mL to about 700 mg / mL. In certain embodiments, provided herein are methods for treating non-Hodgkin's lymphoma (NHL) in a subject in need thereof, comprising administering to the subject a pharmaceutically effective amount of a composition comprising a plurality of particles suspended in a low-viscosity pharmaceutically acceptable liquid carrier, wherein each particle comprises rituximab; the particles have less than about 10% aggregates of the therapeutic biological agent or salt thereof; and the concentration of the therapeutic biological agent or salt thereof in the composition is from about 20 mg / mL to about 700 mg / mL.

[0043] The terms "treat," or "treating," or "treatment" generally refer to both therapeutic treatment and prophylactic or preventative measures, e.g., treating, preventing, reversing, and / or downregulating a disease or condition. As used herein, "treat" or "treating" means administering, internally or externally, a composition containing a therapeutic biological agent, e.g., any of the antibodies or antigen-binding fragments thereof of the present disclosure, to a subject having one or more symptoms or suspected of having a disease for which the biological agent has therapeutic or preventative activity. Typically, a therapeutic biological agent is administered in an amount effective to alleviate one or more symptoms in the treated subject or population by inducing any clinically measurable degree of regression of such symptoms or inhibiting the progression of such symptoms. The amount of a therapeutic biological agent effective to alleviate any particular symptom may vary according to factors such as the condition, age, and weight of the subject or patient and the ability of the therapeutic biological agent to induce a desired response in the subject or patient. Alleviation of disease symptoms can typically be assessed by any clinical measure used by a physician or other skilled healthcare provider to assess the severity or progression of a condition. The terms further include postponing the onset of symptoms associated with a disorder and / or reducing the severity of symptoms of such a disorder. These terms further include ameliorating existing uncontrolled or undesirable symptoms, preventing further symptoms, and ameliorating or preventing the underlying cause of such symptoms. As such, these terms indicate that a beneficial result is achieved in a human or animal subject having a disorder, disease, or condition, or having the potential to develop such a disorder, disease, or condition. As used herein, "treatment," as applied to a human or veterinary subject, refers to therapeutic treatment and diagnostic applications. As applied to a human or veterinary subject, "treatment" encompasses contacting a human or animal subject with a therapeutic biological agent, such as an antibody or antigen-binding fragment of the present disclosure. Subjects in need of treatment for cancer include those already with a benign, precancerous, or non-metastatic tumor and those for preventing the appearance or recurrence of cancer.In some embodiments, the treating or goal or result of treatment may be to reduce the number of cancer cells; reduce the size of the primary tumor; inhibit (i.e., slow to some extent, and preferably stop) cancer cell invasion into peripheral organs; inhibit (i.e., slow to some extent, and preferably stop) tumor metastasis; inhibit tumor growth to some extent; and / or alleviate to some extent one or more symptoms associated with the disorder. In some embodiments, the efficacy of treatment may be measured by assessing the duration of survival, time to disease progression, response rate (RR), duration of response, and / or quality of life. The concentration of the therapeutic biological agent in the composition is typically about 20 mg / mL to about 700 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, 650, 675 mg / mL to about 700 mg / mL. The therapeutic biological agent in the composition can have about 0.5 to about 1.0 unit activity, about 0.75 to about 1.0 unit activity, or about 0.9 to about 1.0 unit activity. The activity is determined by the particle form. The activity is measured relative to the same therapeutic biological agent prior to development. In preferred embodiments, the therapeutic biological agent has a unit activity of about 0.5 to about 1.0. The terms "prevent," or "preventing," or "prevention," as used herein, refer to any action that inhibits or delays the onset of a disease or condition in a subject in need thereof, including administering to the subject a pharmaceutically effective amount of a composition according to the present disclosure. In certain embodiments of the present disclosure, "treating" a subject suffering from a disease or condition includes, but is not limited to, (i) slowing, halting, or reversing the progression of the disease or condition; (ii) slowing, halting, or reversing the progression of symptoms of the disease or condition; (iii) reducing the likelihood of recurrence of the disease or condition; and / or (iv) reducing the likelihood of symptoms of the disease or condition recurring.

[0044] "Precancerous" or "pre-neoplastic" generally refers to a condition or growth that typically precedes or develops into cancer. A "precancerous" growth may have cells characterized by aberrant cell cycle regulation, proliferation, or differentiation, which can be determined by cell cycle markers.

[0045] Examples of cancer include, but are not limited to, blastoma (including medulloblastoma and retinoblastoma), sarcoma (including liposarcoma and synovial cell sarcoma), neuroendocrine tumors (including carcinoid tumor, gastrinoma, and islet cell carcinoma), mesothelioma, schwannoma (including acoustic neuroma), meningioma, adenocarcinoma, melanoma, leukemia or lymphoid malignancy, small cell lung cancer (SGLG), non-small cell lung cancer (NSGLG), adenocarcinoma of the lung, and pulmonary carcinoma. These include lung cancer including squamous cell carcinoma, cancer of the peritoneum, hepatocellular carcinoma, stomach cancer or gastric cancer including gastrointestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer (including metastatic breast cancer), colon cancer, rectal cancer, colorectal cancer, endometrial cancer or uterine cancer, salivary gland cancer, kidney cancer or renal cancer, prostate cancer, vulvar cancer, thyroid cancer, liver cancer, anal cancer, penile cancer, testicular cancer, esophageal cancer, bile duct tumors and head and neck cancer.

[0046] In some embodiments, the "condition or symptom associated with" can be any condition that results from, precedes, or arises from cancer. For example, if the cancer is skin cancer, the condition or associated symptom can be a microbial infection. If the cancer is a secondary tumor, the condition or symptom can relate to organ dysfunction of an associated organ with tumor metastasis. In other embodiments, the methods of treatment described herein are for minimizing or treating a condition or symptom in an individual that is associated with cancer in the individual. In certain embodiments, the subject is a mammal. In preferred embodiments, the subject is a human.

[0047] Non-limiting examples of cancers that can be treated with the compositions and methods described herein include, but are not limited to, tumors of the gastrointestinal tract (colon cancer, rectal cancer, colorectal carcinoma, colorectal cancer, colorectal adenoma, hereditary nonpolyposis 1, hereditary nonpolyposis 2, hereditary nonpolyposis 3, hereditary nonpolyposis 6; colorectal cancer, hereditary nonpolyposis 7, small intestine and / or large intestine cancer, esophageal cancer, callus formation associated with esophageal cancer, stomach carcinoma, pancreatic cancer, pancreatic endocrine tumors), endometrial cancer, dermatofibrosarcoma protuberans, gallbladder cancer, bile duct tumor, prostate cancer, prostate adenocarcinoma, kidney cancer (e.g., Wilms' tumor type 2 or type 1), liver cancer (e.g., hepatoblastoma, hepatocellular carcinoma, carcinoma), hepatocellular carcinoma, bladder cancer, embryonal rhabdomyosarcoma, germ cell tumors, trophoblastic tumors, testicular germ cell tumors, ovary, uterus, immature teratoma of the epithelial ovary, sacrococcygeal tumors, choriocarcinoma, placental trophoblastic tumors, epithelial adult tumors, ovarian cancer, serous ovarian cancer, ovarian sex cord tumors, cervical cancer, small cell and non-small cell lung cancer, nasopharynx, breast cancer (e.g., ductal carcinoma, invasive intraductal carcinoma, sporadic; breast cancer, susceptibility to breast cancer, type 4 breast cancer, breast cancer-1, breast cancer-3; breast-ovarian cancer), squamous cell carcinoma (e.g., in the head and neck), Neuroblastoma, astrocytoma, ganglioneuroblastoma, neuroblastoma, lymphoma (e.g., Hodgkin's disease, non-Hodgkin's lymphoma, B-cell, Burkitt's, cutaneous T-cell, histiocytic, lymphoblastic, T-cell, thymic, B-cell non-Hodgkin's lymphoma), glioma, adenocarcinoma, adrenal gland tumor, hereditary adrenocortical carcinoma, brain malignancies (tumors), various other carcinomas (e.g., bronchogenic large cell, ductal, malignant ascites, Ehrlich-Lettlet ascites, epidermoid, large cell, Lewis lung, medullary, mucoepidermoid, oat cell, small cell, spindle cell, spinous cell, transitional cell, undifferentiated, carcinosarcoma, choriocarcinoma, cyst) adenocarcinoma), ependymoblastoma, epithelioma, erythroleukemia (e.g., Friend, lymphoblastic), fibrosarcoma, giant cell tumor, glial tumor, glioblastoma (e.g., pleomorphic, astrocytoma), glioma, hepatoma, heterohybridoma, heteromyeloma, histiocytoma, hybridoma (e.g., B cell), adrenal gland tumor, insulinoma, pancreatic islet tumor, keratoma, epithelioid leiomyoma, leiomyosarcoma, leukemia (e.g., acute lymphocytic, acute lymphoblastic, acute lymphoblastic pre-B cell, acute lymphoblastic T-cell leukemia, acute megakaryoblastic, monocytic, acute myelogenous, acute myeloid leukemia, acute myelogenous myeloid), acute myeloid with eosinophilia, B cell, basophilic, chronic myeloid, chronic, B cell, eosinophilic, Friend, granulocytic or myeloid, hairy cell, lymphocytic, megakaryoblastic, monocytic, monocytic-macrophage, myeloblastic, myeloid, myelomonocytic, plasma cell, pre-B cell, promyelocytic, subacute, T cell, lymphoid neoplasm, predisposition to myeloid malignancies, acute non-lymphocytic leukemia), lymphosarcoma, melanoma, breast tumor, mast cell tumor, medulloblastoma, mesothelioma, metastatic tumor, monocytic tumor, multiple myeloma, bone myelodysplastic syndrome, myeloma, nephroblastoma, neural tissue glial tumor, neural tissue neuronal tumor, schwannoma, neuroblastoma, oligodendroglioma, osteochondroma, bone myeloma, osteosarcoma (e.g., Ewing), osteoporosis, bone metastasis, papilloma, transitional cell, pheochromocytoma, pituitary tumor (invasive), plasmacytoma, retinoblastoma, rhabdomyosarcoma, sarcoma (e.g., Ewing, histiocytic, Jensen, osteogenic, reticular cell), schwannoma, subcutaneous tumor, teratocarcinoma (e.g., pluripotent), teratoma, testicular tumor, thymoma and trichoepithelioma, gastric cancer, fibrosarcoma, glioblastoma multiforme;Any solid or non-solid cancer and / or cancer metastasis, including, but not limited to, Turcot's syndrome with multiple glomus tumors, Li-Fraumeni syndrome, liposarcoma, Lynch familial cancer syndrome type II, male germ cell tumors, mast cell leukemia, medullary thyroid, multiple meningiomas, endocrine neoplasia, myxosarcoma, paraganglioma, familial nonchromaffin, pilomatricoma, papillary, familial and sporadic, rhabdoid predisposition syndrome, familial, rhabdoid tumor, soft tissue sarcoma, or glioblastoma;

[0048] According to some embodiments of the present disclosure, the compositions and methods described herein further comprise an anti-cancer agent suitable for treatment, including, but not limited to, chemotherapy, biological therapy, photodynamic therapy, nutritional therapy, brachytherapy, immunotherapy, or cell therapy. In some embodiments, the anti-cancer agent can kill cancer cells (cytotoxic agents). In other embodiments, the anti-cancer agent can prevent cancer cells from proliferating (cytostatic agents).

[0049] In some embodiments, the compositions and methods described herein further comprise at least one pharmaceutically acceptable cytotoxic or cytostatic agent. In other embodiments, the cytotoxic or cytostatic agent includes, but is not limited to, abiraterone acetate, Abitrexate (methotrexate), Adriamycin (doxorubicin hydrochloride), Adrucil (fluorouracil), afatinib dimaleate, Afinitor (everolimus), aldesleukin, Alimta (pemetrexed disodium), Aloxi (palonosetron hydrochloride), Ambochlorin (chlorambucil), aminolevulinic acid, anastrozole, aprepitant, Aredia (pamidronate disodium), Arimidex (anastrozole), Aromasin (exemestane), Arranon (nelarabine), arsenic trioxide, asparaginase, blackleg disease (Erwinia chrysanthemi), axitinib, azacitidine, Becenum (carmustine), Beleoodaq (belinostat), bendamustine hydrochloride, bexarotene, bicalutamide, BiC NU (carmustine), bleomycin, bortezomib, Bosulif (bosutinib), bosutinib, busulfan, Busulfex (busulfan), cabazitaxel, cabozantinib-S-malate, Camptosar (irinotecan hydrochloride), capecitabine, carboplatin, carboplatin-TAXOL, carfilzomib, Carmubris (carmustine), carmustine, Casodex (bical tamide), CeeNU (lomustine), ceritinib, Cerubidine (daunorubicin hydrochloride), chlorambucil, chlorambucil-prednisone, cisplatin, Clafen (cyclophosphamide), clofarabine, Clofarex (clofarabine), Clolar (clofarabine), Cometriq (cabozantinib-S-malate), Cosmegen (dactinomycin), crizotinib, cyclophosphamide Rofosfamide, Cyfos (ifosfamide), cytarabine, Cytosar-U (cytarabine), Cytoxan (cyclophosphamide), dabrafenib, dacarbazine, Dacogen (decitabine), dactinomycin, dasatinib, daunorubicin hydrochloride, decitabine, degarelix, denileukin diftitox, dexrazoxane hydrochloride, docetaxel, doxorubicin hydrochloride, DTIC-Dome (da Carbazine), Efudex (fluorouracil), Elitek (rasburicase), Ellence (epirubicin hydrochloride), Eloxatin (oxaliplatin), eltrombopag olamine, Emend (aprepitant), enzalutamide, epirubicin hydrochloride, eribulin mesylate, Erivedge (vismodegib), erlotinib hydrochloride, Erwinaze (asparaginase), blackleg disease (Erwinia chrysanthemi), Etopophos (etoposide phosphate), etoposide, etoposide phosphate, everolimus, Evista (raloxifene hydrochloride), exemestane, Fareston (toremifene), Farydak (panobinostat), Faslodex (fulvestrant), Femara (letrozole), filgrastim, Fludara (fludarabine phosphate), fludarabine phosphate, Fluoroplex (fluorouracil), fluorouracil, Folex (methotrexate), Folex.PFS (methotrexate), Folfirinox, FOLFOX, Folotyn (pralatrexate), fulvestrant, gefitinib, gemcitabine hydrochloride, gemcitabine-cisplatin, gemcitabine-oxaliplatin, Gemzar (gemcitabine hydrochloride), Gilotrif (afatinib dimaleate), Gleevec (imatinib mesylate), Gliadel (carmustine implant), glucarpidase, goserelin acetate, Halaven (eribulin mesylate), Hycamtin (topotecan hydrochloride), Ibr ance (palbociclib), ibrutinib, Iclusig (ponatinib hydrochloride), Idamycin (idarubicin hydrochloride), idarubicin hydrochloride, idelalisib, Ifex (ifosfamide), ifosfamide, Ifosfamidum (ifosfamide), imatinib mesylate, Imbruvica (ibrutinib), Inlyta (axitinib), Iressa (gefitinib), irinotecan hydrochloride, Istodax (romidepsin), ixabepilone, Ixempra (ixabepilone), Jakafi (ruxolitinib phosphate), Jev Tana (cabazitaxel), Keoxifen (raloxifene hydrochloride), Kyprolis (carfilzomib), lanreotide acetate, lapatinib ditosylate, lenalidomide, lenvatinib mesylate, Lenvima (lenvatinib mesylate), letrozole, leucovorin calcium, Leukeran (chlorambucil), leuprolide acetate, Levulan (aminolevulinic acid), Linfolizin (chlorambucil), lomustine, Lupron (leuprolide acetate), Lynparza (olaparib), Marqibo (vinyl sulfate) Cristine liposome), Matulane (procarbazine hydrochloride), mechlorethamine hydrochloride, Megace (megestrol acetate), megestrol acetate, Mekinist (trametinib), mercaptopurine, mesna, Mesnex (mesna), Methazolastone (temozolomide), methotrexate, methotrexate LPF (methotrexate), Mexate (methotrexate), Mexate-AQ (methotrexate), mitomycin C, mitoxantrone hydrochloride, Mitozytrex (mitomycin C), Mozobil (plelixafor), Mustargen (mechlorethamine hydrochloride), Mutamycin (mitomycin C), Myleran (busulfan), Mylosar (azacytidine), Navelbine (vinorelbine tartrate), nelarabine, Neosar (cyclophosphamide), Nexavar (sorafenibut tosylate), nilotinib, Nolvadex (tamoxifen citrate), Nplate (romiplostim), Olapari , omacetaxine mepesuccinate, Ontak (denileukin diftitox), oxaliplatin, paclitaxel, palbociclib, palonosetron hydrochloride, pamidronate disodium, panobinostat, Paraplat (carboplatin), Paraplatin (carboplatin), pazopanib hydrochloride, pemetrexed disodium, Platinol (cisplatin), Platinol-AQ (cisplatin), Plerixa foll, pomalidomide, Pomalyst (pomalidomide), ponatinib hydrochloride, pralatrexate, prednisone, procarbazine hydrochloride, Promacta (eltrombopag olamine), Purinethol (mercaptopurine), Purixan (mercaptopurine), radium-223 dichloride, raloxifene hydrochloride, regorafenib, Revlimid (lenalidomide), Rheumatrex (methotrexate), Midepsin, romiplostim, rubidomycin (daunorubicin hydrochloride), ruxolitinib phosphate, sorafenib tosylate, Sprycel (dasatinib), Stivarga (regorafenib), sunitinib malate, Sutent (sunitinib malate), Synovir (thalidomide), Synribo (omacetaxine mepesuccinate), Tafinlar (dabrafenib), tamoxifen citrate, TarabinePFS (cytarabine), Tarceva (erlotinib hydrochloride), Targretin (bexarotene), Tasigna (nilotinib), Taxol (paclitaxel), Taxotere (docetaxel), Temodar (temozolomide), temozolomide, temsirolimus, thalidomide, Thalomid (thalidomide), thiotepa, Toposar (etoposide), topotecan hydrochloride, toremifene, Torisel (temsirolimus), Totect ( dexrazoxane hydrochloride), trametinib, Treanda (bendamustine hydrochloride), Trisenox (arsenic trioxide), Tykerb (lapatinib ditosylate), vandetanib, Velban (vinblastine sulfate), Velcade (bortezomib), Velsar (vinblastine sulfate), vemurafenib, VePesid (etoposide), Viadur (leuprolide acetate), Vidaza (azacitidine), vinblastine sulfate, Vincasar These include PFS (vincristine sulfate), vincristine sulfate, vinorelbine tartrate, vismodegib, vorinostat, Votrient (pazopanib hydrochloride), Wellcovorin (leucovorin calcium), Xalkori (crizotinib), Xeloda (capecitabine), Xofigo (radium-223 dichloride), Xtandi (enzalutamide), Zaltrap (Ziv-aflibercept), Zelboraf (vemurafenib), Zinecard (dexrazoxane hydrochloride), Ziv-aflibercept, Zoladex (goserelin acetate), zoledronic acid, Zolinza (vorinostat), Zometa (zoledronic acid), Zydelig (idelalisib), Zykadia (ceritinib), Zytiga (abiraterone acetate) or combinations thereof. In a preferred embodiment, the cancer is non-Hodgkin's lymphoma (NHL).

[0050] As described herein, CD20 (also known as Bp35) is a differentiation antigen restricted to B lymphocytes that is expressed during early pre-B cell development and persists until plasma cell differentiation. Because this antigen is expressed at very high density on the surface of malignant B cells, i.e., B cells whose unabated proliferation can lead to B cell lymphoma, CD20 may be a useful target for B cell lymphoma. The U.S. Food and Drug Administration (FDA) has approved the therapeutic use of the anti-CD20 antibody, rituximab (RITUXAN®), for use in relapsed and previously treated indolent non-Hodgkin's lymphoma (NHL). Rituximab acts by binding to the CD20 antigen on B cells, thereby mediating complement-dependent cytotoxicity (CDC). Lysis of B cells results through a mechanism that appears to involve antibody-dependent cell-mediated cytotoxicity (CDC) and antibody-dependent cell-mediated cytotoxicity (ADCC). In certain embodiments, the therapeutic biological agent is an antibody. In a preferred embodiment, the antibody is an anti-CD20 antibody.

[0051] In some embodiments, the therapeutic biological agent is an immunotherapy. In other embodiments, the immunotherapy is an anti-CD20 antibody. In a specific embodiment, the anti-CD20 antibody is rituximab. Preferred embodiments of the compositions and methods described herein may be useful for treating non-Hodgkin's lymphoma (NHL) in a subject in need thereof, comprising administering to the subject a pharmaceutically effective amount of a composition comprising a plurality of particles suspended in a low-viscosity pharmaceutically acceptable liquid carrier, wherein each particle comprises rituximab; the particles have less than about 10% aggregates of the therapeutic biological agent or salt thereof; and the concentration of the therapeutic biological agent or salt thereof in the composition is from about 20 mg / mL to about 700 mg / mL. As described herein, any antibody capable of binding the CD20 antigen may be used in the methods of the present disclosure. Antibodies that bind the CD20 antigen include, for example, C2B8 (rituximab; RITUXAN®) (U.S. Pat. No. 5,736,137, expressly incorporated herein by reference); yttrium-

[90] -labeled 2138 murine antibody, designated Y2B8 (U.S. Pat. No. 5,736,137, expressly incorporated herein by reference); murine IgG2a131 (BEXXAR™), optionally labeled with 1311 to generate the 1311-B1 antibody (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 International Leukocyte Typing Monoclonal antibodies include monoclonal antibodies L27, G28-2, 93-1133, B--Cl, or NU--B2 available from the Workshop (Valentine et al., In: Leukocyte Typing III (McMichael, Ed., p. 440, Oxford University Press (1987)).

[0052] In a specific embodiment of the present disclosure, the anti-CD20 antibody is rituximab. Rituximab is a genetically engineered chimeric mouse / human monoclonal antibody. Rituximab is an IgG, kappa immunoglobulin containing mouse 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, for example, from Genentech (South San Francisco, CA).

[0053] In other embodiments, the anti-CD20 antibodies used in the present disclosure may be administered in conjunction with standard chemotherapy agents / combinations, such as the CHOP chemotherapy regimen, which is a regimen consisting of a combination of cyclophosphamide, doxorubicin, vincristine, and prednisolone. Rituximab has been approved in combination with CHOP chemotherapy for the treatment of certain types of lymphoma, and this combination has become known as RCHOP chemotherapy. In certain embodiments, the composition further comprises at least one of cyclophosphamide, doxorubicin, vincristine, or prednisolone.

[0054] In certain embodiments, the antibodies described herein are antibodies that bind to tumor cells, such as antibodies against cell surface receptors or tumor-associated antigens (TAA). In attempts to discover effective cellular targets for cancer therapy, those skilled in the art have sought to identify transmembrane polypeptides or other tumor-associated polypeptides that are differentially expressed on the surface of one or more specific types of cancer cells compared to one or more normal, non-cancerous cells. In many cases, such tumor-associated polypeptides are more abundantly expressed on the surface of cancer cells compared to the surface of non-cancerous cells. Such cell surface receptors and tumor-associated antigens are well known in the art. and can be prepared for use in producing antibodies using methods and information known in the art.

[0055] Exemplary examples of cell surface receptors and TAAs that can be targeted by the antibodies described herein include, but are not limited to, the various receptors and TAAs listed in the National Center for Biotechnology Information (NCBI) Nucleic Acid and Protein Sequence Identification Code. The nucleic acid and protein sequences corresponding to any listed cell surface receptor and TAA are available in public databases, such as GenBank, and are known in the art.

[0056] Also provided herein is a method of treating an inflammatory disease and / or pain condition in a subject in need thereof, comprising administering to the subject a pharmaceutically effective amount of a composition comprising a plurality of particles suspended in a low-viscosity pharmaceutically acceptable liquid carrier, wherein each particle comprises at least one therapeutic biological agent or salt thereof; the particles have less than about 10% aggregates of the therapeutic biological agent or salt thereof; and the concentration of the therapeutic biological agent or salt thereof in the composition is from about 20 mg / mL to about 700 mg / mL. In certain embodiments, the method further comprises a nonsteroidal anti-inflammatory drug (NSAID). In yet other embodiments, the NSAID may include, but is not limited to, salicylates, indomethacin, flurbiprofen, diclofenac, ketorolac, naproxen, piroxicam, tebufelone, ibuprofen, etodolac, nabumetone, tenidap, alcofenac, antipyrine, aminopyrine, dipyrone, aminopyrone, phenylbutazone, clofezone, oxyphenbutazone, prenazone, apazone, benzydamine, bucolome, cinchophen, clonixin, ditrazol, epizole, fenoprofen, floctafenine, flufenamic acid, glafenine, indoprofen, ketoprofen, loxoprofen, meclofenamic acid, mefenamic acid, niflumic acid, phenacetin, salidifamide, sulindac, suprofen, tolmetin, pharmaceutically acceptable salts thereof, or mixtures thereof.

[0057] In some embodiments of any of the methods described herein, the inflammatory disease or condition includes, but is not limited to, arthritic disease, ophthalmologic disease, retinal disease, Crohn's disease, irritable bowel syndrome, Sjogren's disease, tissue graft rejection, asthma, multiple sclerosis, scleroderma, Goodpasture's syndrome, atherosclerosis, chronic idiopathic thrombocytopenic purpura, Addison's disease, Parkinson's disease, Alzheimer's disease, diabetes, septic shock, myasthenia gravis, inflammatory pelvic disease, inflammatory bowel disease, urethritis, uveitis, sinusitis, pneumonitis, encephalitis, meningitis, myocarditis, chronic osteomyelitis, myositis, hepatitis, gastritis, enteritis, appendicitis, pancreatitis, or cholecystitis.

[0058] In other embodiments, the inflammatory disease or condition is a joint disease. In some embodiments, the joint disease includes, but is not limited to, osteoarthritis, rheumatoid arthritis, spondyloarthritis, systemic juvenile idiopathic arthritis, psoriatic arthritis, gout, ankylosing spondylitis, or juvenile rheumatoid arthritis. In certain embodiments, the subject has previously been diagnosed with an inflammatory condition.

[0059] In some embodiments, the methods provided herein are methods of reducing inflammation, pain, or fever in a subject in need thereof. In other embodiments, pain includes, but is not limited to, nociceptive pain, neuropathic pain, psychogenic pain, breakthrough pain, incident pain, back pain, musculoskeletal pain, postoperative pain, surgical pain, visceral pain, arthritis pain, acute pain, inflammatory pain, or chronic pain. In certain embodiments, the inflammation reduced is in the subject's joints. In yet other embodiments of any of the methods described herein, the subject has a joint disease.

[0060] In some embodiments, inflammatory and / or painful diseases or conditions include, but are not limited to, arthritic pain, musculoskeletal pain, back pain, neuropathic pain, nociceptive pain, acute pain, chronic pain inflammatory pain, surgical pain, postoperative pain, visceral pain, associated pain, breakthrough pain, psychogenic pain, Crohn's disease, irritable bowel syndrome (IBS), inflammatory bowel disease (IBD), Sjogren's disease, tissue graft rejection, asthma, multiple sclerosis, scleroderma, Goodpasture's syndrome, atherosclerosis, Addison's disease, Parkinson's disease, Alzheimer's disease, diabetes, septic shock, myasthenia gravis, inflammatory pelvic disease, inflammatory bowel disease, urethritis, uveitis, sinusitis, pneumonitis, encephalitis, meningitis, myocarditis, chronic osteomyelitis, myositis, hepatitis, gastritis, enteritis, appendicitis, pancreatitis, or cholecystitis. In some examples, the inflammatory condition is a joint disease. Non-limiting examples of joint diseases include osteoarthritis, rheumatoid arthritis, spondyloarthritis, systemic juvenile idiopathic arthritis, psoriatic arthritis, gout, ankylosing spondylitis or juvenile rheumatoid arthritis.

[0061] In other embodiments, the inflammatory and / or painful disease or condition is a chronic or acute inflammatory process, hi certain embodiments, the chronic or acute inflammatory process may be rheumatoid arthritis, Crohn's disease, ulcerative colitis, or sepsis.

[0062] In some embodiments, the inflammatory and / or painful disease or condition includes, but is not limited to, metabolic syndrome or disease, ulcerative colitis, Crohn's disease, irritable bowel syndrome (IBS), or inflammatory bowel disease (IBD). In other embodiments, the inflammatory bowel disease (IBD) is Crohn's disease of the small intestine (CDSB), Crohn's disease of the colon (CDC), or ulcerative colitis (UC). In particular embodiments, the metabolic syndrome or disease is associated with inflammation, such as type II diabetes, obesity, or fatty acid metabolism disorders.

[0063] In other embodiments, the compositions and methods described herein are useful for treating immune disorders, acquired hypogammaglobulinemia secondary to hematologic malignancies, chronic inflammatory demyelinating polyneuropathy (CIDP), Guillain-Barré syndrome, idiopathic thrombocytopenic purpura, inflammatory myopathies, Lambert-Eaton myasthenic syndrome, multifocal motor neuropathy, myasthenia gravis, Mersch-Wortmann syndrome, secondary hypogammaglobulinemia, specific antibody deficiencies, acute disseminated encephalomyelitis, autoimmune hemolytic anemia; cicatricial pemphigoid, It is useful to treat Evans syndrome, feto-maternal / neonatal alloimmune thrombocytopenia (FMAIT / NAIT), hemophagocytic syndrome, high-risk allogeneic hematopoietic stem cell transplant, IgM paraproteinemic neuropathy, kidney transplant, multiple sclerosis, opsoclonus-myoclonus ataxia, post-transfusion purpura, toxic epidermal necrolysis / Stevens-Johnson syndrome (TEN / SJS), toxic shock syndrome, Alzheimer's disease, multiple myeloma, sepsis; B-cell neoplasms, trauma, or bacterial, viral, or fungal infections.

[0064] In some embodiments of any of the methods described herein, the immune disease is common variable immunodeficiency (CVID), congenital agammaglobulinemia, Wiskott-Aldrick syndrome, severe combined immunodeficiency (SCID), primary hypogammaglobulinemia, primary immunodeficiency with antibody deficiency, sex-linked recessive agammaglobulinemia (XLA), infantile hypogammaglobulinemia, or paraneoplastic cerebellar degeneration without antibodies.

[0065] In other embodiments, the immune disease is an autoimmune disease. In certain embodiments, the autoimmune disease includes, but is not limited to, multiple sclerosis, scleroderma, type 1 diabetes, rheumatoid arthritis, thyroiditis, Raynaud's syndrome, Sjogren's syndrome, autoimmune uveitis, autoimmune myocarditis, inflammatory bowel disease, amyotrophic lateral sclerosis (ALS), systemic lupus, neuromyelitis optica, idiopathic thrombocytopenic purpura, myasthenia gravis, ulcerative colitis, Crohn's disease, polyarthritis, graft-versus-host reaction, juvenile-onset diabetes, Hashimoto's thyroiditis, Graves' disease, pernicious anemia, chronic active (lupoid) hepatitis, psoriatic arthritis, or neurodermatitis.

[0066] In certain embodiments, immune disorders include low antibody levels, impaired function of white blood cells due to problems with T lymphocytes, problems with T lymphocytes, problems with B lymphocytes, death and dysfunction of white blood cells, and abnormalities. In other embodiments, the infection can be classified as an immune disorder, i.e., common variable immunodeficiency, selective antibody deficiency, transient hypogammaglobulinemia of infancy, X-linked recessive agammaglobulinemia (Bruton's agammaglobulinemia), chronic mucocutaneous candidiasis, DiGeorge anomaly, ataxia-telangiectasia, severe combined immunodeficiency, Wiskott-Aldrick syndrome, X-linked lymphoproliferative syndrome, human immunodeficiency virus, Chediak-Higashi syndrome, leukocyte glucose-6-phosphate dehydrogenase deficiency, myeloperoxidase, hyperglobulinemia E (Jobe-Buckles syndrome), leukocyte adhesion defect, complement component 3 (C3) deficiency, complement component 6 (C6) deficiency, complement component 7 (C7) deficiency, or complement component 8 (C8) deficiency.

[0067] In other embodiments, the methods described herein further comprise administering to the patient a therapeutic agent. In certain embodiments, the therapeutic agent is an anti-inflammatory agent, an antihistamine, an analgesic, or a corticosteroid. In yet other embodiments, the methods further comprise administering a nonsteroidal anti-inflammatory drug (NSAID). In some embodiments, the NSAID can be salicylate, indomethacin, flurbiprofen, diclofenac, ketorolac, naproxen, piroxicam, tebufelone, ibuprofen, etodolac, nabumetone, tenidap, alcofenac, antipyrine, aminopyrine, dipyrone, aminopyrone, phenylbutazone, clofezone, oxyphenbutazone, prenazone, apazone, benzydamine, bucolome, cinchophen, clonixin, ditrazol, epizole, fenoprofen, floctafenine, flufenamic acid, glafenine, indoprofen, ketoprofen, loxoprofen, meclofenamic acid, mefenamic acid, niflumic acid, phenacetin, salidifamide, sulindac, suprofen, tolmetin, a pharmaceutically acceptable salt thereof, or a mixture thereof.

[0068] In some embodiments, the method comprises treating a tumor in a tumor selected from the group consisting of neuroblastoma, sarcoma, brain cancer, metastatic brain cancer, ovarian cancer, prostate cancer and breast cancer, lymphoma, colorectal cancer, metastatic colorectal cancer, colorectal cancer, colorectal tumors, non-small cell lung cancer, carcinoma, blood cancer, hematological malignancies, hematological tumors, gastrointestinal cancer, non-Hodgkin's lymphoma, Hodgkin's lymphoma, B-cell non-Hodgkin's lymphoma, metastatic cancer, squamous cell carcinoma, malignant ascites, gastric cancer, head and neck cancer, squamous cell carcinoma of the head and neck, solid tumors, pancreatic cancer, bone metastasis, recurrent glioblastoma multiforme, Malignant melanoma, melanoma, multiple myeloma, cancers expressing Nectin-4, chronic lymphocytic leukemia, adrenocortical carcinoma, non-small cell lung cancer, B-cell lymphoma, B-cell carcinoma, acute myeloid leukemia, renal clear cell carcinoma, renal cell carcinoma, gastrointestinal adenocarcinoma, pancreatic tumors, gastric cancer, nasopharyngeal carcinoma, glioma, acute lymphoblastic leukemia, follicular lymphoma, T-cell lymphoma, platelet aggregation inhibition, inflammatory bowel disease, ulcerative colitis, Crohn's disease, Clostridium difficile difficile colitis, rheumatoid arthritis, plaque psoriatic arthritis, ankylosing spondylitis, juvenile idiopathic arthritis, hemolytic disease of the newborn, multiple sclerosis, amyotrophic lateral sclerosis, hypercholesterolemia, asthma, allergic asthma, chronic asthma, severe allergic disorders, allergic reactions, inflammatory lesions, inflammation, retinopathy of prematurity, thromboembolism, thrombosis, oncology / immunology indications, treatment of autoimmune diseases, invasive Candida infections, reduction of scarring after glaucoma surgery, white blood cell disorders, systemic sclerosis, Bacillus anthracis spores, anthrax (prophylaxis and treatment), immunologically mediated inflammatory disorders, immunodeficiency disorders, Pseudomonas aeruginosa aeruginosa infection, tumor necrosis factor, reduction of side effects of cardiac surgery, macular degeneration (wet form), neovascular age-related macular degeneration, recovery of motor function after stroke, inflammation of the airway, non-malignant skin cancer, inflammation of the gastrointestinal tract, uveitis, polymyositis, choroidal and retinal neovascularization, muscular dystrophy, orthopedic muscle atrophy, sarcopenia, pain, Escherichia coli (E.Diarrhea caused by Staphylococcus aureus (coli), Alzheimer's disease, sepsis, sepsis caused by gram-negative bacteria, sepsis (Staphylococcus), viral infections, osteoporosis, chronic osteomyelitis, bleeding, infections / influenza A, atopic diseases, paroxysmal nocturnal hemoglobinuria, heart attack, stroke, traumatic shock, hepatitis B, chronic hepatitis B. The method is useful for treating appendicitis, acute sciatica, respiratory syncytial virus infection, respiratory syncytial virus prophylaxis, rabies (prophylaxis), idiopathic pulmonary fibrosis, pain, HIV infection, cytomegalovirus infection, hemorrhagic shock, fibrosis, percutaneous coronary intervention, Staphylococcus aureus infection, prevention of organ transplant rejection, graft-versus-host disease, reversal of the anticoagulant effect of dabigatran, antineoplastic agent, myostatin inhibitor, dyslipidemia, diabetes, diabetes mellitus type 1, cardiac imaging, diagnostic agent, or tumor detection. In other embodiments, the organ transplant rejection (host-versus-graft reaction) is prophylaxis for transplant rejection. In certain embodiments, the method is useful for treating non-malignant skin cancer. In a preferred embodiment, the method is useful for treating non-malignant skin cancer.

[0069] Provided herein is a method of treating a renal disease in a subject in need thereof, comprising administering to the subject a pharmaceutically effective amount of a composition comprising a plurality of particles suspended in a low viscosity pharmaceutically acceptable liquid carrier, wherein each particle comprises at least one therapeutic biological agent or salt thereof; the particles have less than about 10% aggregates of the therapeutic biological agent or salt thereof; and the concentration of the therapeutic biological agent or salt thereof in the composition is from about 20 mg / mL to about 700 mg / mL. In certain embodiments, provided herein is a method of treating membranoproliferative glomerulonephritis (MPGN), membranoproliferative glomerulonephritis (MPGN) pathogenesis, focal segmental glomerulosclerosis (FSGS), membranous nephropathy (MN), severe membranous nephropathy (MN), steroid-resistant nephrotic syndrome, vasculitis, or a combination thereof, in a subject in need thereof, comprising administering to the subject a pharmaceutically effective amount of a composition comprising a plurality of particles suspended in a low-viscosity pharmaceutically acceptable liquid carrier, wherein each particle comprises at least one therapeutic biological agent or salt thereof; the particles have less than about 10% aggregates of the therapeutic biological agent or salt thereof; and the concentration of the therapeutic biological agent or salt thereof in the composition is from about 20 mg / mL to about 700 mg / mL.

[0070] "Treatment," in reference to kidney disease, means (1) inhibiting the onset of symptoms of the disease, e.g., causing clinical symptoms of the disease not to occur in an animal (e.g., a human) that has or may be susceptible to developing the disease but has not yet experienced or displayed symptoms of the disease; (2) inhibiting the disease, e.g., arresting the onset of the disease or one or more of its clinical symptoms; or (3) alleviating or ameliorating the disease, e.g., causing regression of the disease or one or more of its clinical symptoms. For example, treatment can refer to alleviating one or more symptoms associated with kidney disease. Treating kidney disease does not require the complete absence of the disease. For example, a reduction of at least 25% or at least 50% of one or more symptoms or undesirable consequences of the disease may be sufficient.

[0071] "Reduce" or "reduce" means to become less or smaller in number, amount, size, or intensity. In certain embodiments, reducing the risk of a disease (e.g., focal segmental glomerulosclerosis (FSGS)) includes reducing the likelihood of developing the disease by at least about 20%, e.g., at least about 30%, 40%, 50%, 60%, 70%, 80%, or 90%. In other embodiments, reducing the risk of a disease includes a delay in the onset of the disease, e.g., a delay of at least about 6 months, e.g., about 1 year, e.g., about 2 years, about 5 years, or about 10 years.

[0072] The term "renal disease" or "kidney disease," as generally used herein, means a disorder that specifically results in kidney damage. Renal diseases include, but are not limited to, focal segmental glomerulosclerosis (FSGS), hypertensive end-stage renal disease (ESRD), nephropathy secondary to systemic lupus erythematosus, diabetic nephropathy, hypertensive nephropathy, IgA nephropathy, IgA nephritis, HIV-associated nephropathy, non-diabetic chronic kidney disease, chronic kidney disease, reflux nephropathy, glomerulonephritis, glomerulonephropathy, polycystic kidney disease, or Kidney disease can be chronic or acute. Chronic kidney disease can progress, in certain embodiments, from stage 1 (slightly reduced kidney function) to stage 2 (mild reduction in function), stage 3 (moderate reduction in function), stage 4 (severe reduction in function), or stage 5 (established kidney failure), as described herein.

[0073] In some embodiments, the "condition or symptom associated with" can be any condition that results from, precedes, or arises from renal disease. For example, if the renal disease is focal segmental glomerulosclerosis (FSGS), the condition or associated symptom can be a microbial infection. In other embodiments, the methods of treatment described herein are for minimizing or treating a condition or symptom in an individual that is associated with renal disease in the individual. In certain embodiments, the subject is a mammal. In preferred embodiments, the subject is a human.

[0074] In some embodiments, the desired response is preventing the onset of kidney disease (e.g., focal segmental glomerulosclerosis (FSGS)). In other embodiments, the desired response is delaying the onset or progression of kidney disease (e.g., focal segmental glomerulosclerosis (FSGS)), e.g., by at least about 3 months, at least about 6 months, at least about 1 year, at least about 2 years, at least about 5 years, or at least about 10 years. In certain embodiments, the desired response is reducing signs and symptoms of kidney disease (e.g., focal segmental glomerulosclerosis (FSGS)), such as scarring of kidney tissue and / or neurological symptoms in the hands and feet or associated with speaking.

[0075] As disclosed herein, the methods can be used to treat kidney diseases. In some embodiments, the kidney disease is antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV), idiopathic membranous nephropathy, membranoproliferative glomerulonephritis (MPGN), lupus nephritis, systemic lupus erythematosus (SLE), minimal change disease (MCD), focal segmental glomerulosclerosis (FSGS), antibody-mediated kidney transplant rejection, membranous nephropathy (MN), severe membranous nephropathy (MN), severe membranous nephropathy (MN), steroid-resistant nephrotic syndrome, vasculitis, IgA nephritis, diabetic nephropathy, chronic kidney disease, reflux nephropathy, glomerulonephritis, glomerular nephropathy, polycystic kidney disease, or as part of a desensitization regimen to allow for ABO / human leukocyte antigen (HLA)-mismatched kidney transplantation. In a preferred embodiment, the method can be used to treat membranoproliferative glomerulonephritis (MPGN), membranoproliferative glomerulonephritis (MPGN) pathogenesis, focal segmental glomerulosclerosis (FSGS), membranous nephropathy (MN), severe membranous nephropathy (MN), steroid-resistant nephrotic syndrome, vasculitis, or a combination thereof.

[0076] In certain embodiments, the methods described herein can significantly reduce kidney damage in individuals with a common form of kidney disease called membranous nephropathy. In preferred embodiments, the methods can be used as a first-line treatment for severe membranous nephropathy (MN).

[0077] In certain embodiments, the methods may show best treatment outcomes in patients with steroid-resistant nephrotic syndrome, particularly those with focal segmental glomerulosclerosis (FSGS) and young patients suffering from recurrent FSGS.

[0078] In some embodiments, the methods described herein can reduce the number of harmful antibodies, called autoantibodies (ANCA), produced by the body by targeting and destroying B cells. ANCA attacks healthy tissues and cells and is produced by B cells. Autoantibodies target specific white blood cells called neutrophils, and ANCA causes neutrophils to stick to and aggregate in the walls of small blood vessels in different tissues and organs of the body. In other embodiments, the methods can be used to treat vasculitis, which can harm the kidneys.

[0079] In certain other embodiments, the methods can be used to treat or reduce the likelihood of developing focal segmental glomerulosclerosis (FSGS) in subjects who have not had a kidney transplant or in subjects identified as being at risk for developing kidney disease.

[0080] In some embodiments, the subject is an end-stage renal disease (ESRD) patient, a patient undergoing immunosuppressive therapy, an AIDS patient, a diabetic patient, a newborn, an organ transplant patient, a patient with a compromised immune system, the elderly, a patient with an autoimmune disease, a burn patient, a cancer patient, or a patient in an acute care setting.

[0081] The methods as described herein may be administered in combination with other known therapies for the treatment of kidney disease. In some embodiments, the composition further comprises at least one therapeutic agent. For example, a subject treated with a composition of the present disclosure may also be treated with a blood pressure medication, a steroid, and / or an immunosuppressant. Examples of therapeutic agents include blood pressure medications (e.g., diuretics (e.g., chlorthalidone, chlorothiazide, furosemide, hydrochlorothiazide, indapamide, metolazone, amiloride hydrochloride, spironolactone, triamterene, bumetanide, or a combination thereof), alpha adrenergic antagonists (e.g., alfuzosin, doxazosin, prazosin, terazosin, or tamsulosin, or a combination thereof), central adrenergic inhibitors (e.g., clonidine, guanfacine, or or methyldopa or a combination thereof), angiotensin-converting enzyme (ACE) inhibitors (e.g., benazepril, captopril, enalapril, fosinopril, lisinopril, moexipril, perindopril, quinapril, ramipril, or trandolapril or a combination thereof), angiotensin II receptor blockers (e.g., candesartan, eprosartan, irbesartan, losartan, olmesartan, telmisartan, or valsartan or a combination thereof), alfa α-blockers (e.g., doxazosin, prazosin, or terazosin, or a combination thereof), beta-blockers (e.g., acebutolol, atenolol, betaxolol, bisoprolol, carteolol, metoprolol, nadolol, nebivolol, penbutolol, pindolol, propranolol, solotol, or timolol, or a combination thereof), calcium channel blockers (e.g., amlodipine, bepridil, diltiazem, felodipine, isradipine, nicardipine, benzodiazepine, nifedipine, nisoldipine or verapamil or a combination thereof), vasodilators (e.g., hydralazine or minoxidil or a combination thereof) and renin inhibitors (e.g., aliskiren) or a combination thereof), steroids (e.g., corticosteroids such as cortisone, prednisone, methylprednisolone or prednisolone) or anabolic steroids (anatrophin, anakisval, anadrol, bolasterone, decadiabolin, deca-durabolin,Dehydroepiandrosterone (DHEA), delatestril, dianiabol, dihydrolone, durabolin, dimethazine, enoltestobis, expose, gamma hydroxybutyric acid, maxibolin, metatriol, methyltestosterone, paraboline, primobolin, quinolones, serabolin, trophobolene and winstrol) or immunosuppressants, such as glucocorticoids, cytostatics, antibodies or anti-inflammatory drugs. The composition may comprise at least one of the following: munofilin and / or mycophenolate mofetil (MMF), FK-506, azathioprine, cyclophosphamide, methotrexate, dactinomycin, antithymocyte globulin (ATGAM), muromonab-CD3 antibody, basilizimab, daclizumab, cyclosporine, tacrolimus, voclosporin, sirolimus, interferon, infliximab, etanercept, adalimumab, fingolimod, and / or myriocin. In a preferred embodiment, the composition further comprises at least one of a steroid, diphenhydramine, acetaminophen, a blood pressure medication, an immunosuppressant, or a combination thereof.

[0082] Provided herein is a pharmaceutical composition comprising a pharmaceutical effective amount of a low viscosity, pharmaceutically acceptable liquid carrier. Also provided is a method of treating a skin disease or condition in a subject in need thereof, comprising administering to the subject a composition comprising a plurality of particles suspended in the body, each particle comprising at least one therapeutic biological agent or salt thereof; the particles having less than about 10% aggregates of the therapeutic biological agent or salt thereof; and the concentration of the therapeutic biological agent or salt thereof in the composition is from about 20 mg / mL to about 700 mg / mL. In certain embodiments, the method further comprises a nonsteroidal anti-inflammatory drug (NSAID). In yet other embodiments, the NSAID may include, but is not limited to, salicylates, indomethacin, flurbiprofen, diclofenac, ketorolac, naproxen, piroxicam, tebufelone, ibuprofen, etodolac, nabumetone, tenidap, alcofenac, antipyrine, aminopyrine, dipyrone, aminopyrone, phenylbutazone, clofezone, oxyphenbutazone, prenazone, apazone, benzydamine, bucolome, cinchophen, clonixin, ditrazol, epizole, fenoprofen, floctafenine, flufenamic acid, glafenine, indoprofen, ketoprofen, loxoprofen, meclofenamic acid, mefenamic acid, niflumic acid, phenacetin, salidifamide, sulindac, suprofen, tolmetin, pharmaceutically acceptable salts thereof, or mixtures thereof.

[0083] The methods as described herein may be administered in combination with other known therapies for the treatment of skin disorders. In some embodiments, the composition further comprises at least one therapeutic agent. For example, a subject treated with a composition of the present disclosure may also be treated with a blood pressure medication, a steroid, and / or an immunosuppressant. Examples of therapeutic agents include blood pressure medications (e.g., diuretics (e.g., chlorthalidone, chlorothiazide, furosemide, hydrochlorothiazide, indapamide, metolazone, amiloride hydrochloride, spironolactone, triamterene, bumetanide, or a combination thereof), alpha adrenergic antagonists (e.g., alfuzosin, doxazosin, prazosin, terazosin, or tamsulosin, or a combination thereof), central adrenergic inhibitors (e.g., clonidine, guanfacine, or or methyldopa or a combination thereof), angiotensin-converting enzyme (ACE) inhibitors (e.g., benazepril, captopril, enalapril, fosinopril, lisinopril, moexipril, perindopril, quinapril, ramipril, or trandolapril or a combination thereof), angiotensin II receptor blockers (e.g., candesartan, eprosartan, irbesartan, losartan, olmesartan, telmisartan, or valsartan or a combination thereof), alfa α-blockers (e.g., doxazosin, prazosin, or terazosin, or a combination thereof), beta-blockers (e.g., acebutolol, atenolol, betaxolol, bisoprolol, carteolol, metoprolol, nadolol, nebivolol, penbutolol, pindolol, propranolol, solotol, or timolol, or a combination thereof), calcium channel blockers (e.g., amlodipine, bepridil, diltiazem, felodipine, isradipine, nicardipine, benzodiazepine, nifedipine, nisoldipine or verapamil or a combination thereof), vasodilators (e.g., hydralazine or minoxidil or a combination thereof) and renin inhibitors (e.g., aliskiren) or a combination thereof), steroids (e.g., corticosteroids such as cortisone, prednisone, methylprednisolone or prednisolone) or anabolic steroids (anatrophin, anakisval, anadrol, bolasterone, decadiabolin, deca-durabolin,dehydroepiandrosterone (DHEA), delatestril, dianiabol, dihydrolone, durabolin, dimethazine, enoltestobis, expose, gamma hydroxybutyric acid, maxibolin, metatriol, methyltestosterone, paraboline, primobolin, quinolones, serabolin, trophobolene and winstrol) or immunosuppressants, such as glucocorticoids, cytostatics, antibodies or anti-immunophilins and / or mycophenolate mofetil (MMF), FK-506, azathioprine, cyclophosphamide, methotrexate, dactinomycin, antithymocyte globulin (ATGAM), muromonab-CD3 antibody, basilizimab, daclizumab, cyclosporine, tacrolimus, voclosporin, sirolimus, interferon, infliximab, etanercept, a, The compositions include rituximab, dalimumab, fingolimod, and / or myriocin. In a preferred embodiment, the compositions further comprise at least one of a steroid, diphenhydramine, acetaminophen, a blood pressure medication, an immunosuppressant, or a combination thereof. In particular, the U.S. Food and Drug Administration (FDA) has approved the use of rituximab in combination with systemic steroids for the treatment of granulomatosis with polyangiitis and microscopic polyangiitis in adult patients. In a preferred embodiment, the compositions further comprise at least one steroid.

[0084] The term "skin disease" or "skin condition," as generally used herein, means a disorder that specifically results in damage to the skin. Skin diseases include, but are not limited to, dermatitis, contact dermatitis, eczema, urticaria, rosacea, cicatricial psoriatic lesions of the skin, psoriasis, atopic dermatitis, contact hypersensitivity, acne, dermatomyositis, vitiligo, ichthyosis vulgaris, photoallergic hypersensitivity, infantile hemangiomas, common warts, psoriasis, neurofibromatosis, epidermolysis bullosa, atopic eczema, food allergies, granulomatosis with polyangiitis (e.g., Wegener's granulomatosis), cutaneous vasculitis (e.g., acute hemorrhagic edema, Henoch-Schönlein purpura, hypersensitivity vasculitis), genital ulcers, and genital ulcers. These include pemphigus vulgaris, dermatomyositis, atopic dermatitis (e.g., eczema), lupus erythematosus or primary bullous diseases associated with autoantibodies against various structural support proteins in the epidermis and dermo-epidermal junction (e.g., autoimmune bullous disorders), pemphigus vulgaris and treatment-resistant cases (e.g., pemphigus foliaceus, paraneoplastic pemphigus associated with CD20+ lymphoma, bullous pemphigoid, epidermolysis bullosa acquisita, mucous membrane pemphigoid), refractory cutaneous lupus erythematosus, thrombotic thrombocytopenic purpura, or a combination thereof.

[0085] In some embodiments, the subject in need or aware of a need for a treatment as described herein may be a patient suffering from, by way of example only, dermatitis, contact dermatitis, eczema, urticaria, rosacea, cicatricial psoriatic lesions of the skin, psoriasis, atopic dermatitis, contact hypersensitivity, acne, dermatomyositis, vitiligo vulgaris, ichthyosis vulgaris, photoallergic hypersensitivity, infantile hemangiomas, common warts, psoriasis, neurofibromatosis, epidermolysis bullosa, atopic eczema, food allergies, granulomatosis with polyangiitis (e.g., Wegener's granulomatosis), cutaneous vasculitis (e.g., acute hemorrhagic edema, Henoch-Schönlein purpura), or other skin conditions, including, but not limited to, skin allergies, skin allergies, skin allergies, skin allergies, skin sensitivities ... The patient suffers from a skin disease including pemphigus vulgaris and treatment-resistant cases (e.g., pemphigus foliaceus, paraneoplastic pemphigus associated with CD20+ lymphoma, bullous pemphigoid, epidermolysis bullosa acquisita, mucous membrane pemphigoid), refractory cutaneous lupus erythematosus, thrombotic thrombocytopenic purpura, or a combination thereof.

[0086] In other embodiments, the skin disease is granulomatosis with polyangiitis (e.g., Wegener's granulomatosis), cutaneous vasculitis (e.g., acute hemorrhagic edema, Henoch-Schönlein purpura, hypersensitivity vasculitis), primary bullous disease, dermatomyositis, atopic dermatitis (e.g., eczema), lupus erythematosus or primary bullous disease associated with autoantibodies against various structural support proteins at the epidermis and dermo-epidermal junction (e.g., autoimmune bullous disorders), pemphigus vulgaris and treatment-resistant cases (e.g., pemphigus foliaceus, paraneoplastic pemphigus associated with CD20+ lymphoma, bullous pemphigoid, epidermolysis bullosa acquisita, mucous membrane pemphigoid), refractory cutaneous lupus erythematosus, thrombotic thrombocytopenic purpura, or a combination thereof.

[0087] In certain embodiments, the skin disease is granulomatosis with polyangiitis, e.g., Wegener's granulomatosis or microscopic polyangiitis. Granulomatosis with polyangiitis is a potentially fatal form of vasculitis in which there is an immune response in which, for example, antibodies damage the walls of small blood vessels and surrounding tissue. In certain embodiments, patients with granulomatosis with polyangiitis suffer from skin ulcers, e.g., palpable purpura, nodules, papules and vesicles, pyoderma gangrenosum, Raynaud's phenomenon, or a combination thereof.

[0088] In certain other embodiments, the skin disease is cutaneous vasculitis. Cutaneous vasculitis is a group of disorders in which inflamed blood vessels, such as capillaries, venules, arterioles, and lymphatic vessels, are present in the skin. Classifications of cutaneous vasculitis include, but are not limited to, capillaritis, such as progressive pigmented purpura (the most common form of capillaritis), pruritic purpura, pigmented purpuric lichenoid dermatosis, telangiectatic purpura annulare, capillaritis associated with contact allergy, pigmented purpuric dermatosis; small-vessel vasculitis, such as idiopathic, drug- or infection-induced cutaneous small-vessel vasculitis (hypersensitivity vasculitis), Henoch-Schönlein purpura, acute hemorrhagic edema of infancy, urticarial vasculitis, exercise-induced vasculitis, and vasculitis. These include inflammatory vasculitis, erythema elevatum elevata, malignant atrophic papulosis (Degos), cryoglobulinemia, recurrent cutaneous necrotizing eosinophilic vasculitis, ANCA-associated vasculitis, microscopic polyangiitis, eosinophilic granulomatosis with polyangiitis / Churg-Strauss, granulomatosis with polyangiitis, lymphomatoid granulomatosis; medium-sized vasculitis, cutaneous polyarteritis nodosa, Kawasaki disease, vasculitis nodosa; and large-sized vasculitis, such as temporal arteritis, Takayasu's disease, and giant cell arteritis. Vasculitis is classified as a type III hypersensitivity reaction involving immune complexes, i.e., antibodies bound to antigens in affected blood vessels. In another embodiment, the skin disease is atopic dermatitis. Atopic dermatitis (eczema) results from complex interactions between various immune cells and proteins and is characterized by high levels of serum IgE in many patients.

[0089] Human immunodeficiency virus (HIV) affects specific cells of the immune system called CD4 cells or T cells. Over time, HIV can destroy these cells to the point that the body is unable to fight off infection and disease. HIV disease has a well-documented progression. Untreated, HIV is almost universally fatal as it eventually overwhelms the immune system, leading to acquired immune deficiency syndrome (AIDS). HIV treatment can help people at all stages of the disease and slow or prevent progression from one stage to the next. As is known in the art, the clinical signs and symptoms of HIV infection are primarily caused by the proliferation of lymphocytes marked by the CD3 and CD4 antigens (CD 4 + T cells) This is due to the significant loss of life expectancy. It is also generally accepted that the infectious agent in AIDS is HIV. Non-limiting examples of symptoms of HIV infection within the first few weeks following exposure to the virus include fever, headache, sore throat, swollen lymph nodes, and rash. Additional symptoms of HIV infection that may appear later include mild infection, swollen lymph nodes, diarrhea, weight loss, fever, cough, and shortness of breath. HIV infection typically progresses to AIDS in about 10 years. By the time AIDS develops, the subject's immune system is severely damaged. Late-stage symptoms of HIV infection or AIDS include profuse night sweats, weeks of shaking chills or fever above 38°C, cough or shortness of breath, chronic diarrhea, persistent white spots or unusual lesions on the tongue or in the mouth, headache, persistent unexplained fatigue, blurred or distorted vision, weight loss, and rash or bumps. Subjects with HIV infection or AIDS also have an increased risk of developing several opportunistic infections, such as bacterial, viral, fungal and protozoan infections, as well as cancer. Non-limiting examples of opportunistic bacterial infections include bacterial strains that cause diarrhea (e.g., Salmonella sp., Campylobacter sp., Shigella sp.), bacterial strains that cause pneumonia (e.g., Streptococcus pneumoniae, Haemophilus influenzae, Pseudomonas aeruginosa, Staphylococcus aureus, Legionella pneumophila, Mycoplasma pneumoniae, and Chlamydia pneumoniae), Mycobacterium avium, Treponema pallidum, and the like. Non-limiting examples of opportunistic viral infections include cytomegalovirus (CMV), hepatitis C, herpes simplex virus, and flu-like viruses. (oral and genital herpes), varicella zoster virus (shingles), human papillomavirus (HPV), molluscum contagiosum virus (MCV), Epstein-Barr virus (EBV), and JC polyomavirus. Non-limiting examples of opportunistic fungal infections include Aspergillus, Candida albicans, Coccidioides immitis, Coccidioides posadasii, Cryptococcus neoformans, and Histoplasma capsulatum. Non-limiting examples of opportunistic protozoan infections include Cryptosporidium species, Isospora belli, Microsporidium species (e.g., Enterocytozoon bieneusi), Pneumocystis jiroveci, and Toxoplasma gondii. Subjects with HIV infection or AIDS also have an increased risk of developing cancer, including, but not limited to, anal dysplasia or cancer, cervical dysplasia or cancer, Kaposi's sarcoma, and lymphoma.

[0090] As used herein, the term "human immunodeficiency virus (HIV) infection" refers to the presence of at least one HIV virion in a subject, a detectable level of HIV virions in a subject, or the presence or detectable level of HIV genomic nucleic acid in a subject. Non-limiting examples of methods for detecting the presence or level of HIV virions or HIV genomic nucleic acid in a subject are known in the art. For example, methods for detecting or measuring the level of HIV genomic nucleic acid in a subject, such as reverse transcriptase polymerase chain reaction, are known in the art. Methods for indirectly detecting the presence of HIV virions, such as measuring anti-HIV antibody titers in a subject, are also known in the art. With respect to human immunodeficiency virus (HIV) infection, "reducing" refers to a decrease in number, amount, or level. For example, reducing HIV viral load refers to a reduction or decrease in the amount of HIV in relevant bodily fluids. A reduction can generally be compared to an initial starting number, amount, or level, but can also be compared to a control or standard number, amount, or level.

[0091] In some embodiments, compounds that inhibit HIV infection of CD4 T cells include, for example, entry inhibitors such as CC chemokine receptor type 5 (CCR5) inhibitors, CXX chemokine receptor type 4 (CXCR4) inhibitors, CD4 inhibitors, gp120 inhibitors, and gp41 inhibitors (e.g., enfuvirtide); and anti-inflammatory drugs such as hydroxychloroquine, chloroquine, PD-1 inhibitors, type I interferons, IL6, cyclooxygenase 2 inhibitors, peroxisome proliferator-activated receptor-c (PPAR-c) agonists (e.g., pioglitazone and leflunomide), methotrexate, mesalazine, and anti-fibrotic agents (e.g., angiotensin-converting enzyme (ACE) inhibitors). Examples of CCR5 inhibitors include maraviroc, aplaviroc, and vicriviroc. Other examples of entry inhibitors include TNX-355, PRO140, BMS-488043, plerixafor, epigallocatechin gallate, anti-gp120 antibodies such as antibody b12, Griffithsin, DCM205, and designed ankyrin repeat proteins (DARPins). HAART is used to reduce the chance that the virus will develop resistance. Compounds for HAART are well known and include, for example, combinations of two or more nucleoside reverse transcriptase inhibitors (NRTIs), such as tenofovir, emtricitabine, zidovudine (AZT), lamivudine (3TC), abacavir, and tenofovir alafenamide fumarate; and one or more non-nucleotide reverse transcriptase inhibitors (NNRTIs), such as efavirenz, rilpivirine, and etravirine; integrase inhibitors, such as raltegravir and elvitegravir; and / or protease inhibitors, such as ritonavir, These include darunavir, atazanavir, lopinavir, and cobicistat. The HAART drugs most frequently used to treat HIV infection include nucleoside / nucleotide reverse transcriptase inhibitors, such as tenofovir, emtricitabine, and abacavir; and non-nucleoside reverse transcriptase inhibitors (NNRTIs), such as efavirenz, nevirapine, or etravirine; protease inhibitors (PIs), such as atazanavir, ritonavir, or darunavir; fusion and entry inhibitors, such as enfuvirtide and maraviroc; and integrase inhibitors, such as raltegravir. In other embodiments, compounds that stimulate reactivation of latent HIV include, for example, histone deacetylase (HDAC) inhibitors, such as vorinostat, pomidepsin, panobinostat, gibinostat, belinstat, valproic acid, CI-994, MS-275, BML-210, M344, NVP-LAQ824, mocetinostat, and sirtuin inhibitors; NF-κB inducers, such as anti-CD3 / CD28 antibodies, tumor necrosis factor alpha (TNFα), prostratin, ionomycin, bryostatin- 1 and picologs; histone methyltransferase (HMT) inhibitors, such as BIX-01294 and chaetocin; proapoptotic and cell differentiation molecules, such as JQ1, nutlin 3, disulfiram, aphidicolin, hexamethylene bisacetamide (HMBA), dactinomycin, aclarubicin, cytarabine, Wnt small molecule inhibitors, and Notch inhibitors; immunomodulators, such as anti-PD-1 antibodies, anti-CTLA-4 antibodies, anti-TRIM-3 antibodies, and BMS-936558; and CD4 T cell vaccines. Combinations of such stimulating agents can also be used. The effect of some stimulating agents on HIV reactivation can also be enhanced by combining them with other compounds.

[0092] In other embodiments, "symptoms of HIV infection" refer to any measurable or detectable physical manifestation of HIV infection or AIDS in a human subject. One or more symptoms of HIV infection can be detected during a physical examination by a medical professional. Non-limiting examples of symptoms of HIV infection in humans include fever, headache, muscle and joint pain, sore throat, rash, diarrhea, swollen lymph nodes, e.g., in the neck, axilla, or groin, night sweats, loss of appetite, fatigue, weight loss, dry cough, shortness of breath, depression, peripheral neuropathy, confusion, altered level of consciousness, lesions on the tongue or in the mouth, blurred or distorted vision, or a combination thereof. In certain embodiments, "symptoms of HIV infection" also include the presentation of one or more opportunistic infections in a subject, e.g., bacterial, viral, fungal, and / or parasitic infections. Opportunistic infections are typically not observed in human subjects with healthy immune systems, e.g., immunocompromised subjects.Non-limiting examples of opportunistic infections include cytomegalovirus (CMV) infection, Pneumocystis carinii pneumonia (PCP) (Pneumocystis jirovecii), Mycobacterium tuberculosis infection, recurrent pneumonia, candidiasis infection (thrush), coccidioidomycosis, cryptococcosis, herpes simplex virus infection, e.g., herpes simplex virus-1 infection, Salmonella infection, Shigella infection, Listeria infection, Campylobacter infection, cryptosporidiosis, microsporidiosis, Mycobacterium avium complex infection, and the like. These include: HIV / AIDS (Hodgkin's Virus or Mycobacterium Angusii), astrovirus, histoplasmosis, isosporosis, adenovirus infection, rotavirus infection, Clostridium difficile infection, toxoplasmosis (Toxoplasma gondii), Cryptococcus neoformans infection, Penicillium marneffei infection, or a combination thereof.

[0093] In some embodiments, a plurality of therapeutic biological agents comprising at least one therapeutic biological agent or salt thereof. Administration of a composition comprising the particles reduces opportunistic infections in a subject with HIV infection or AIDS, or reduces the risk of developing an opportunistic infection in a subject with HIV infection or AIDS. For example, the methods described herein can result in a substantial reduction in the number or severity, frequency, or duration of one or more symptoms of an opportunistic bacterial, fungal, viral, or parasitic infection, such as any of the exemplary opportunistic bacterial, fungal, viral, or parasitic infections described herein, in a subject with HIV infection or AIDS who has such an opportunistic infection. In other embodiments, the methods described herein reduce the risk of developing an opportunistic bacterial, viral, fungal, or parasitic infection, such as any of the exemplary opportunistic bacterial, fungal, viral, or parasitic infections described herein, in a subject with HIV infection or AIDS who does not have an opportunistic bacterial, viral, fungal, or parasitic infection, e.g., compared to a subject or group of subjects with HIV infection or AIDS who have not received treatment or an alternative form of treatment. In still other embodiments, the methods described herein reduce the risk of developing cancer, e.g., any of the exemplary cancers described herein, in a subject with HIV infection or AIDS, e.g., compared to a subject or group of subjects with HIV infection or AIDS who are not receiving treatment or an alternative form of treatment. In certain other embodiments, administering a composition comprising a plurality of particles suspended in a low-viscosity pharmaceutically acceptable liquid carrier, each particle comprising at least one therapeutic biological agent or salt thereof, results in a reduction, e.g., a significant or detectable reduction, e.g., at least about a 5%, 10%, 15%, 20%, 25%, or about a 30% reduction, in HIV titer in the subject.

[0094] Pharmaceutical Composition In certain embodiments, the present disclosure relates to compositions, e.g., pharmaceutical compositions, comprising a plurality of particles suspended in a low viscosity pharmaceutically acceptable liquid carrier, each particle comprising any one of the therapeutic biological agents described above.

[0095] In one aspect, the present disclosure provides a composition, e.g., a pharmaceutically effective composition, comprising a plurality of particles suspended in a low viscosity pharmaceutically acceptable liquid carrier, wherein each particle comprises at least one therapeutic biological agent or salt thereof; the particles have less than about 10% aggregates of the therapeutic biological agent or salt thereof; and the concentration of the therapeutic biological agent or salt thereof in the composition is from about 20 mg / mL to about 700 mg / mL.

[0096] Patients, including but not limited to humans, can be treated by administering to the patient an effective amount of a pharmaceutical composition comprising an active therapeutic biological agent or a pharmaceutically acceptable prodrug or salt thereof in the presence of a pharmaceutically acceptable liquid, carrier, or diluent. The active material can be administered in liquid form by any suitable route, for example, parenterally, intradermally, or subcutaneously. Parenteral dosage forms are intended for administration as an injection. Common types of injection are intravenous (into a vein), subcutaneous (under the skin), and intramuscular (into a muscle). Infusions are typically given by the intravenous route.

[0097] The concentration of the active therapeutic biological agent in the composition will depend on the absorption, inactivation, and excretion rates of the drug, as well as other factors known to those skilled in the art. It should be noted that dosage values ​​will also vary depending on the severity of the condition to be alleviated and potential drug-drug interactions with antiretroviral drugs. For any particular subject, specific dosage regimens should be adjusted over time according to the individual's needs and the professional judgment of the person administering or supervising the administration of the composition, and it is further understood that the concentration ranges described herein are merely exemplary and are not intended to limit the scope or practice of the claimed compositions. The active ingredient can be administered immediately, or can be divided into several smaller doses to be administered at varying time intervals.

[0098] The therapeutic biological agent or its pharmaceutically acceptable prodrug or salt can also be mixed with other additives or active ingredients that do not impair the desired action, or with ingredients that supplement the desired action, such as antibiotics, antifungals, anti-inflammatory agents, or other antivirals, including, but not limited to, nucleoside compounds. Solutions, suspensions, or compositions used herein for intradermal or subcutaneous administration can contain the following components: a sterile diluent, such as water for injection, saline, fixed oils, polyethylene glycol, glycerin, propylene glycol, or other synthetic solvents; an antibacterial agent, such as benzyl alcohol or methylparaben; an antioxidant, such as ascorbic acid or sodium bisulfite; a chelating agent, such as ethylenediaminetetraacetic acid; a buffer, such as acetic acid, citric acid, histidine, succinic acid, phosphate, or a combination thereof, and an agent for regulating osmotic pressure, such as sodium chloride or dextrose, or a combination thereof. Parenteral preparations can be enclosed in disposable syringes or cartridges made of glass, plastic, or the like. In certain embodiments, the composition is dispensed from a pre-filled syringe.

[0099] The compositions and methods of the present disclosure can be used to treat a subject in need thereof. In certain embodiments, the subject is a mammal, such as a human or a non-human mammal. When administered to a mammal, such as a human, the composition is preferably administered as a pharmaceutical composition, for example, comprising the composition of the present disclosure and a pharmaceutically acceptable liquid carrier. Pharmaceutically acceptable liquid carriers are known in the art and include, for example, aqueous solutions, such as water or physiologically buffered saline, or other solvents or vehicles, such as glycols, glycerol, oils, such as olive oil, or injectable organic esters. In preferred embodiments, when such pharmaceutical compositions are intended for human administration, for example, parenteral administration, the aqueous solution is pyrogen-free or substantially pyrogen-free. In certain preferred embodiments, the pharmaceutically acceptable liquid carrier is non-aqueous.

[0100] In some embodiments, the pharmaceutically acceptable carrier can contain a physiologically acceptable agent that acts, for example, to stabilize or increase the absorption of the therapeutic biological agent of the present disclosure. Such physiologically acceptable agents include, for example, carbohydrates such as glucose, sucrose, trehalose, or dextran; antioxidants such as ascorbic acid or glutathione; chelating agents; low molecular weight proteins; or other stabilizers or excipients. The choice of a pharmaceutically acceptable carrier containing a physiologically acceptable agent depends, for example, on the route of administration of the composition. In a preferred embodiment, the composition is administered by subcutaneous syringe injection.

[0101] The phrase "pharmaceutically acceptable" is used herein to refer to those therapeutic biological agents, materials, compositions, and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans 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 therapeutic biological agents and compositions that do not produce adverse, allergic, or other untoward reactions when administered to a mammal, e.g., a human, as appropriate. The preparation of pharmaceutical compositions containing antibodies or additional active ingredients will be known to those of skill in the art in light of the present disclosure. Moreover, it will be understood that for mammalian, e.g., human, administration, preparations should meet sterility, pyrogenicity, general safety, and purity standards as required by the FDA Office of Biological Standards.

[0102] The phrase "pharmaceutically acceptable carrier" as used herein means a pharmaceutically acceptable material, composition, or vehicle, such as a liquid, diluent, excipient, or solvent. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the composition and not harmful to the patient. Some examples of materials that can serve as pharmaceutically acceptable carriers include, but are not limited to, (1) sugars, such as lactose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; (4) tragacanth powder; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository wax; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; (10) glycols, such as propylene Glycols; (11) polyols, such as glycerin, sorbitol, mannitol, and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) phosphate buffer solution; and (21) other non-toxic, compatible substances used in pharmaceutical compositions. Those skilled in the art may dilute or increase the volume of therapeutic biological agents with inert materials. These diluents may include carbohydrates, particularly trehalose, mannitol, α-lactose, anhydrous lactose, cellulose, sucrose, modified dextrans, and starch. Certain inorganic salts, including calcium triphosphate, magnesium carbonate, and sodium chloride, may also be used as fillers. Some commercially available diluents are Fast-Flo, Emdex, STA-Rx1500, Emcompress, and Avicell. The phrase "pharmaceutically acceptable carrier" includes any and all aqueous solvents (e.g., water, alcoholic / aqueous solutions, saline, parenteral vehicles such as sodium chloride, Ringer's dextrose, and the like), non-aqueous solvents (e.g., propylene glycol, polyethylene glycol, vegetable oils, and injectable organic esters such as ethyl oleate), dispersion media, coatings, surfactants, antioxidants, preservatives (e.g., antibacterial or antifungal agents, antioxidants, chelating agents, and inert gases), isotonic agents, absorption delaying agents, salts, drugs, drug stabilizers, gels, binders, excipients, disintegrants, lubricants, sweeteners, flavorings, dyes, fluid and nutrient replenishers, such materials, and combinations thereof, as are known to those skilled in the art.The pH and exact concentration of the various components in the pharmaceutical composition are controlled by well-known parameters.

[0103] In some embodiments, pharmaceutical compositions or formulations for parenteral administration comprise aqueous solutions of the active therapeutic biological agent in water-soluble form. Preferably, suspensions of the active therapeutic biological agent may be prepared as appropriate oily injectable compositions. Suitable lipophilic solvents or vehicles include fatty oils (e.g., sesame oil, corn oil), or synthetic fatty acid esters (e.g., ethyl oleate or triglycerides), or liposomes. Optionally, the composition may also contain suitable stabilizers or agents that increase the solubility of the therapeutic biological agent to allow for the preparation of highly concentrated solutions. Pharmaceutical compositions may also contain solvents or dispersion media containing, for example, water, ethanol, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), suitable mixtures thereof, and vegetable oils. Proper fluidity can be maintained by maintaining the required particle size in the case of dispersions and by the use of surfactants. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In certain cases, it may be preferable to include isotonic agents, for example, sugars or sodium chloride.

[0104] Suitable buffering agents include acetic acid and salts (1-2% w / v); citric acid and salts (1-3% w / v); boric acid and salts (0.5-2.5%) w / v); or phosphoric acid and salts (0.8-2% w / v). Suitable preservatives include benzalkonium chloride (0.003-0.03%) w / v; chlorobutanol (0.3-0.9%) w / v; parabens (0.01-0.25%) w / v; or thimerosal (0.004-0.02%) w / v.

[0105] Pharmaceutical compositions, such as formulations as disclosed herein, can be administered, for example, parenterally (including intramuscularly, intravenously, subcutaneously, or intrathecally, for example, as sterile solutions or suspensions); intraperitoneally; The composition can be administered to a subject by any of several administration routes, including intravenous, intravenous, intraperitoneal, or intravenous. In some embodiments, the composition is administered parenterally, subcutaneously, orally, epidermally, intradermally, intramuscularly, intraarterially, intraperitoneally, or intravenously. In certain embodiments, the composition can be simply suspended in a non-aqueous liquid carrier. Details of suitable administration routes and compositions for these can be found, for example, in 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 and the patents cited therein. The term "pharmaceutical composition" or "formulation," as disclosed herein, refers to a preparation that is in a form that allows a therapeutic biological agent in the composition to be effective, e.g., when administered to a subject, and that does not contain additional components that are unacceptably toxic to the subject to which the composition or formulation is administered. Such a composition or formulation is sterile. A "sterile" composition or formulation is aseptic or free of all living microorganisms and their spores.

[0106] The compositions may be conveniently presented in unit dosage form and prepared by any method known in the art of pharmacy. The amount of active ingredient which can be combined with a liquid carrier material to produce a single dosage form will vary depending on the host treated and the particular mode of administration. The amount of active ingredient which can be combined with a carrier material to produce a single dosage form will generally be that amount of therapeutic biological agent which produces a therapeutic effect. Generally, out of 100 percent, this amount will range from about 1% to about 99% active ingredient, preferably from about 50% to about 99%, and most preferably from about 70% to about 98%.

[0107] In some embodiments of the present disclosure, compositions suitable for use in the present disclosure may be administered parenterally, particularly subcutaneously.

[0108] An effective amount of the composition can be administered in a single dose per day or in divided doses throughout the day, e.g., two to three times daily. By way of example, administration of compositions according to the present disclosure can generally occur over an extended period of at least one week, two weeks, three weeks, four weeks, or even four to fifteen weeks, optionally including one or more rest periods or repeated after rest periods, e.g., three or more times daily. In some embodiments, the composition is administered one or more times daily, e.g., one, two, three, four, five, six, seven, eight, nine, ten, or more times daily. In other embodiments, the composition is administered for about 1 to about 31 days, e.g., 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 22 days, 23 days, 24 days, 25 days, 26 days, 27 days, 28 days, 29 days, 30 days, or 31 days. In certain embodiments, the composition is administered for at least 1 day. In still other embodiments, the composition is administered for one or more weeks, e.g., 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, or more. In yet other embodiments, the composition is administered for one or more months, such as 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months or more.

[0109] In certain embodiments, the therapeutic biological agent may be administered at a dose of about 1 mg to about 1,500 mg per day, about 5 mg to about 1,300 mg per day, about 10 mg to about 900 mg per day, about 20 mg to about 600 mg per day, about 40 mg to about 300 mg per day, about 150 mg to about 350 mg per day, about 40 mg to about 150 mg per day, about 25 mg to about 150 mg per day, about 2.5 mg to about 150 mg per day, about 20 mg to about 80 mg per day, or about 1 mg to about 30 mg per day. In still other embodiments, the therapeutic biological agent may be administered at a dose of about 1,300 mg / day, about 900 mg / day, about 600 mg / day, about 150 mg / day, about 250 mg / day, about 150 mg / day, about 250 mg / day, about 2.5 mg / day, about 150 mg / day, about 20 mg / day, about 80 mg / day, or about 1 mg / day. In some embodiments, the therapeutic biological agent is administered at a dose of about 10 mg to about 1000 mg per day.

[0110] In other embodiments, the measured daily dose for a therapeutic biological agent is from about 0.01 mg / kg per day to about 1500 mg / kg per day for a human subject. Dosages may be adjusted appropriately to achieve desired local or systemic drug levels upon administration. For example, subcutaneous administration is expected to result in doses one to several orders of magnitude higher per day compared to IV infusion. If the subject responds inadequately at such doses, even higher doses (or higher effective doses via more localized delivery routes) may be used, to the extent patient tolerance permits. Multiple doses per day may be contemplated to achieve adequate systemic levels of the therapeutic biological agent.

[0111] For any therapeutic biological agent composition described herein, the therapeutically effective amount can be initially determined from animal models. The therapeutically effective dose can also be determined from human data for therapeutic biological agents of the present disclosure tested in humans and for therapeutic biological agents known to exhibit similar pharmacological activity, such as other related active agents. Higher doses may be required for parenteral administration. The applied dose can be adjusted based on the relative bioavailability and efficacy of the administered therapeutic biological agent. Adjusting the dose to achieve maximum efficacy based on the compositions and methods described above and other methods known in the art is well within the capabilities of one of ordinary skill in the art.

[0112] To facilitate dissolution of the therapeutic biological agent in an aqueous environment, a surfactant may be added as an additive. The surfactant may include an anionic detergent, such as sodium lauryl sulfate, dioctyl sodium sulfosuccinate, or dioctyl sodium sulfonate. Cationic detergents that can be used include benzalkonium chloride or benzethonium chloride. Possible nonionic detergents that can be included in the composition as surfactants include lauromacrogol 400, polyoxyl 40 stearate, polyoxyethylene hydrogenated castor oil 10, 50, and 60, glycerol monostearate, polysorbate 20, 40, 60, 65, and 80, sucrose fatty acid esters, methylcellulose, or carboxymethylcellulose. These surfactants can be present alone or in a mixture of different ratios in the therapeutic biological agent composition or derivative of the present disclosure.

[0113] The present disclosure includes the use of pharmaceutically acceptable salts of therapeutic biological agents in the methods of the present disclosure. The term "pharmaceutically acceptable salt," as used herein, includes salts derived from inorganic or organic acids, including, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, perchloric acid, phosphoric acid, formic acid, acetic acid, lactic acid, maleic acid, fumaric acid, succinic acid, tartaric acid, glycolic acid, salicylic acid, citric acid, methanesulfonic acid, benzenesulfonic acid, benzoic acid, malonic acid, trifluoroacetic acid, trichloroacetic acid, naphthalene-2-sulfonic acid, or other acids. In some embodiments, pharmaceutically acceptable salt forms can include forms in which the ratio of salt-containing molecules is not 1:1. For example, a salt can contain multiple inorganic or organic acid molecules per molecule of base, e.g., two hydrochloric acid molecules per molecule of therapeutic biological agent. As another example, a salt may contain less than one inorganic or organic acid molecule per molecule of base, for example, two molecules of therapeutic biological agent per molecule of tartaric acid.

[0114] In certain embodiments, salts contemplated by the present disclosure include, but are not limited to, arginine, benethamine, benzathine, betaine, calcium hydroxide, choline, deanol, diethanolamine, diethylamine, 2-(diethylamino)ethanol, ethanolamine , ethylenediamine, N-methylglucamine, hydrabamine, 1H-imidazole, L-lysine, magnesium, 4-(2-hydroxyethyl)morpholine, piperazine, potassium, 1-(2-hydroxyethyl)pyrrolidine, sodium, triethanolamine, tromethamine, or zinc salts. In other embodiments, contemplated salts of the present disclosure include, but are not limited to, Na, Ca, K, Mg, Zn, or other metal salts. In further embodiments, contemplated salts of the present disclosure include, but are not limited to, alkyl, dialkyl, trialkyl, or tetraalkylammonium salts. Pharmaceutically acceptable salts may also exist as various solvates, for example, with water, methanol, ethanol, dimethylformamide, and the like. Mixtures of such solvates can also be prepared. The source of such solvates may be from the solvent of crystallization, inherent in the solvent of preparation or crystallization, or exogenous to such solvent.

[0115] In yet another embodiment, the proteinaceous composition may be formulated in a neutral or salt form. Pharmaceutically acceptable salts include acid addition salts (formed with the protein's free amino groups) and are formed with inorganic acids such as hydrochloric or phosphoric acids, or organic acids such as acetic, oxalic, tartaric, mandelic, and the like. Salts formed with free carboxyl groups may also be derived from inorganic bases such as sodium, potassium, ammonium, calcium, or ferric hydroxides, and organic bases such as isopropylamine, trimethylamine, histidine, procaine, and the like.

[0116] As one of skill in the art will appreciate, compositions of the present disclosure that have no adverse effects upon administration to a subject can be administered to a subject daily.

[0117] Preferred embodiments of the present disclosure are described herein. Naturally, variations, modifications, and equivalent substitutions of those preferred embodiments will be apparent to those skilled in the art upon reading the above description. The inventors anticipate that those skilled in the art will employ such variations, modifications, modifications, and equivalent substitutions as appropriate, and the inventors intend for the present disclosure to be practiced other than as specifically described herein. Those skilled in the art will readily recognize a variety of non-critical parameters that can be changed, altered, or modified to yield essentially similar results. Accordingly, the present disclosure includes all modifications and equivalents of the subject matter recited in the claims appended hereto as permitted by applicable law. Moreover, any combination of the above-described elements in all possible variations thereof is encompassed by the present disclosure unless otherwise indicated herein or otherwise clearly contradicted by context.

[0118] Whereas each element of the present disclosure is described herein as including multiple embodiments, it should be understood that, unless otherwise indicated, each embodiment of a given element of the present disclosure can be used with each embodiment of the other elements of the present disclosure, and that each such use is intended to form a separate embodiment of the present disclosure.

[0119] It will be appreciated by those skilled in the relevant art that other suitable modifications and adaptations to the compositions and methods described herein will be readily apparent from the description of the disclosure contained herein, in view of the information known to those skilled in the art, and may be made without departing from the scope of the disclosure or any embodiment thereof.

[0120] definition For purposes of this disclosure, the following definitions will be used unless expressly stated otherwise.

[0121] As used in the context of describing this disclosure, the terms "a," "an," "the," and similar referents are to be construed to encompass both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. All compositions and methods described can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any examples or exemplary language (e.g., "etc.") provided herein is intended merely to better elucidate the disclosure and does not pose a limitation on the scope of the otherwise claimed disclosure. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the disclosure.

[0122] For example, the term "about" with respect to given numerical values ​​for temperature and time period means including numerical values ​​within 10% of the particular value.

[0123] As used herein, the term "administering" refers to the actual physical introduction of a composition into or onto a subject (as appropriate). Any method of introducing a composition into a subject is contemplated by this disclosure. The method is not dependent on any particular means of introduction, and should not be construed as such. Means of introduction are known to those of skill in the art and are exemplified herein.

[0124] As used herein, "amino acid" or "residue" refers to any natural or unnatural amino acid, any amino acid derivative, or any amino acid mimetic known in the art. Included are L- and D-forms of each amino acid, although the L-form is generally preferred. In some embodiments, the term refers to any one of the 20 natural amino acids in their L-form: glycine (Gly), alanine (Ala), valine (Val), leucine (Leu), isoleucine (Ile), 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 certain embodiments, the amino acid side chain can be the side chain of Gly, Ala, Val, Leu, Ile, Met, Cys, Ser, Thr, Trp, Phe, Lys, Arg, His, Tyr, Asn, Gln, Asp, Glu, or Pro.

[0125] As used herein, unless the context requires otherwise, the term "comprise" and variations of this term, such as "comprises," "including," and "including," are not intended to exclude additional additives, components, integers, or steps. The terms "comprises" and "comprising" may be used interchangeably. As used herein, the phrases "selected from the group consisting of," "selected from," and the like, include mixtures of specified materials. When a numerical limit or range is stated herein, the endpoints are included. Also, all values ​​and subranges within the numerical limit or range are specifically included as if expressly written out. Reference to an element in the singular is not intended to mean "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.

[0126] As used herein, the terms "effective amount," "effective dose," "sufficient amount," "effective amount," "therapeutically effective amount," "pharmaceutical effective amount," or grammatical equivalents thereof, refer to a dosage sufficient to produce a desired result, improve, or in some manner reduce symptoms or halt or reverse the progression of a condition, achieving a subjective relief of symptoms or an objectively identifiable improvement as noted by a clinician or other qualified observer. Improvement of symptoms of a particular condition by administration of a pharmaceutical composition described herein refers to a reduction, whether sustained or temporary, permanent or transient, that may be associated with administration of the pharmaceutical composition. As noted above, "effective amount" refers to any amount sufficient to achieve a desired biological effect. By selecting from a variety of active therapeutic biological agents and weighting factors, such as potency, relative bioavailability, patient weight, severity of adverse side effects, and preferred mode of administration, in conjunction with the teachings provided herein, one can design an effective prophylactic or therapeutic treatment regimen that does not result in substantial undesirable toxicity and is effective in treating a particular subject. The effective amount for any particular application can vary depending on factors such as the disease or condition being treated, the particular therapeutic biological agent of the present disclosure being administered, the size of the subject, or the severity of the disease or condition. One of ordinary skill in the art can empirically determine the effective amount of a particular therapeutic biological agent of the present disclosure and / or other therapeutic agent without necessitating undue experimentation. It is generally preferred to use the maximum dose, which is the highest safe dose according to some medical judgment. Multiple doses per day may be intended to achieve an appropriate systemic level of the therapeutic biological agent. The appropriate systemic level can be determined, for example, by measuring peak or sustained plasma levels of the drug in the patient. "Dose" and "administration" are used interchangeably herein. The term "dose" or "dosage" refers to a physically discrete unit suitable for use in a subject, each unit containing a predetermined amount of a therapeutic composition calculated to produce the desired response discussed above in connection with its administration, i.e., appropriate route and treatment regimen. The amount administered, both in terms of number of treatments and doses, depends on the desired effect. The actual amount of the composition of the present embodiments administered to a patient or subject can be determined by physical and physiological factors, such as the subject's weight, age, health, and sex, the type of disease being treated, the degree of disease penetration, previous or concurrent therapeutic interventions, the patient's idiopathic condition, the route of administration, and the efficacy, stability, and toxicity of the particular therapeutic agent. For example, doses can range from about 1 μg / kg / body weight to about 1000 mg / kg / body weight per administration (such ranges including intervening doses) or more, and any range derivable therein. Non-limiting examples of ranges that can be derived from the numbers recited herein include ranges of about 5 μg / kg / body weight to about 100 mg / kg / body weight, about 5 μg / kg / body weight to about 500 mg / kg / body weight, etc.The practitioner responsible for administration will, in any event, determine the concentration of active ingredient(s) in a composition and appropriate dose(s) for the individual subject.

[0127] The term "oligopeptide" is used to refer to peptides with a smaller number of amino acid residues, as opposed to polypeptides or proteins. The oligopeptides described herein typically consist of about 2 to about 40 amino acid residues. Oligopeptides include dipeptides (2 amino acids), tripeptides (3 amino acids), tetrapeptides (4 amino acids), pentapeptides (5 amino acids), hexapeptides (6 amino acids), heptapeptides (7 amino acids), octapeptides (8 amino acids), nonapeptides (9 amino acids), decapeptides (10 amino acids), undecapeptides (11 amino acids), dodecapeptides (12 amino acids), icosapeptides (20 amino acids), tricontapeptides (30 amino acids), and tetracontapeptides (40 amino acids). Oligopeptides can also be classified by molecular structure, such as aeruginosins, cyanopeptolins, microcystins, microviridins, microginins, anabaenopeptins, and cyclamides. Homo-oligopeptides are oligopeptides that contain the same amino acids. In a preferred embodiment, the homo-oligopeptides contain polyvaline, polyalanine, and polyglycine hexamers of 10 amino acids.

[0128] The term "peptide" is defined as a small protein of two or more amino acids linked by the carboxyl group of one amino acid to the amino group of another. Thus, at its basic level, any type of peptide synthesis involves repeated steps of adding amino acids or peptide molecules to each other or to an existing peptide chain. The term "peptide" generally has from about 2 to about 100 amino acids, while a polypeptide or protein has about 100 or more amino acids up to the full-length sequence that can be translated from a gene. Furthermore, as used herein, peptide refers to a polypeptide or protein. can be a subsequence or portion of a protein. In certain embodiments, the peptide is 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 preferred embodiments, the peptide is about 30 to about 100 amino acids in length. In some embodiments, the peptide is about 40 to about 100 amino acids in length.

[0129] As used herein, the term "pharmaceutically acceptable" refers to a composition that is physiologically tolerable and typically does not produce an allergic or similar adverse reaction when administered to a subject, preferably a human subject. Preferably, as used herein, the term "pharmaceutically acceptable" means approved by a federal or state regulatory agency or listed in the United States Pharmacopoeia or other generally recognized pharmacopoeias for use in animals, more particularly in humans.

[0130] As used herein, a treatment that "prevents" a disorder or condition refers to a therapeutic biological agent that, in a statistical sample, reduces or delays the onset of the disorder or condition in a treated sample relative to an untreated control sample, or reduces the severity of one or more symptoms of the disorder or condition in a treated sample relative to an untreated control sample.

[0131] As used herein, the term "prodrug" is intended to encompass therapeutic biological agents that are converted under physiological conditions to the therapeutically active biological agents of the present disclosure. A common method for making prodrugs includes one or more selected moieties that are hydrolyzed under physiological conditions to reveal the desired molecule. In other embodiments, the prodrug is converted by enzymatic activity in the host animal. For example, esters or carbonates, such as esters or carbonates of alcohols or carboxylic acids, are preferred prodrugs of the present disclosure. In certain embodiments, some or all of the molecules in the compositions depicted above can be replaced with the corresponding appropriate prodrug; for example, a hydroxyl in the parent molecule is presented as an ester, or a carbonate or carboxylic acid present in the parent therapeutic biological agent is presented as an ester.

[0132] The term "protein" is defined as a linear polymer of about 20 different amino acids. The types and sequence of amino acids in a protein are specified by the DNA that produces the protein. In certain embodiments, the sequence can be natural and non-natural. The sequence of amino acids determines the overall structure and function of the protein. In some embodiments, a protein can contain 50 or more residues. In preferred embodiments, a protein can contain more than about 101 residues in length. The net charge of a protein can be determined by two factors: 1) the total number of acidic amino acids relative to basic amino acids; and 2) the particular solvent pH environment that exposes positive or negative residues. As used herein, a "net positively or net negatively charged protein" is a protein that has a net positive or net negative charge in a non-denaturing pH environment. Those skilled in the art will understand that, in general, all proteins can be considered "net negatively charged proteins" regardless of their amino acid composition due to their pH and / or solvent environment. For example, different solvents can expose negative or positive side chains depending on the solvent pH. The protein or peptide is preferably selected from any type of enzyme or antibody or fragment thereof that exhibits substantially the same activity as the corresponding enzyme or antibody. The protein or peptide may serve as a structural material, e.g., keratin, an enzyme, a hormone, a transporter, e.g., hemoglobin, an antibody, or a regulator of gene expression. The protein or peptide is required for the structure, function, and regulation of cells, tissues, and organs.

[0133] The term "substantially free," as used herein, refers to a majority or majority, such as at least about 51%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%, 99.99%, or at least about 99.999% or more.

[0134] The term "treating" is art-recognized and includes administration of one or more of the subject compositions to a host, for example to reduce, ameliorate, or stabilize an existing undesirable condition or its side effects.

[0135] It is understood that the specific order or hierarchy of steps in any disclosed method or process is illustrative of a sample approach. Based on design preferences, it is understood that the specific order or hierarchy of steps in a method or process may be rearranged. Some of the steps may occur simultaneously. Other steps may occur continuously or intermittently, for example, with on / off periods occurring at regular or irregular intervals, and the duration of the periods and / or the allocation therebetween may vary in a structured or unstructured manner. The accompanying methods call for the presence of elements of the various steps in a sample order and are not meant to be limited to the specific hierarchy or order presented. The use of a phrase, such as "embodiment," does not imply that such embodiment applies to all configurations of the subject technology. Disclosure regarding an embodiment may apply to all embodiments or to one or more embodiments. A phrase, such as "one embodiment," may refer to one or more embodiments, and vice versa.

[0136] Methods of the present disclosure When clinical applications of therapeutic compositions containing therapeutic biological agents, e.g., mAbs, are attempted, it is generally beneficial to prepare pharmaceutical or therapeutic compositions that are appropriate for the intended use. In certain embodiments, pharmaceutical compositions may contain, for example, at least about 0.1% of an active therapeutic biological agent. In other embodiments, the active therapeutic biological agent may comprise from about 2% to about 99% or from about 50% to about 99% by weight of the unit, and any range derivable therein.

[0137] For large molecule biological agents, such as mAbs, protein instability and high viscosity can result from intermolecular interactions in solution. High viscosity can make the drug composition difficult to handle and inject. Protein instability can reduce the effective dose as less protein is therapeutically useful and can form aggregates that can harm patients.

[0138] As described herein, microparticle-based compositions or suspension formulations can directly address the current challenges of subcutaneous therapeutic biological agent, e.g., mAb, protein, delivery by allowing for high concentrations (>400 mg / mL) of therapeutic biological agents while maintaining an injectable, e.g., injectable, format with excellent mAb or protein stability. The terms "particle" and "microparticle" are used interchangeably herein in their broadest sense. In some embodiments, solid microparticles limit intermolecular interactions responsible for high viscosity and instability in aqueous compositions. In other embodiments, the particles are suspended in a liquid carrier vehicle, preventing dissolution prior to injection. The compositions described herein can be loaded into a pre-filled syringe or injection device format, e.g., needleless injection, and can be administered in a complex manner. This eliminates the need for error-prone reconstitution procedures. The highly dispersible nature of the particles in the compositions described herein allows for easy resuspension by gentle shaking, making subcutaneous injection patient-friendly. Within the subcutaneous space, therapeutic biological agents, including the particles described herein, easily return to their original monomeric state upon injection, allowing for full bioavailability. In a preferred embodiment, the composition achieves higher loading without compromising the quality of the therapeutic biological agent, thus allowing therapeutic biological agents, such as mAbs, to be delivered in pre-filled syringes via subcutaneous injection.

[0139] Provided herein are compositions and methods useful for treating a disease or condition in a subject in need thereof, comprising administering to the subject a composition comprising a pharmaceutically effective amount of a plurality of particles suspended in a low viscosity pharmaceutically acceptable liquid carrier, each particle comprising at least one therapeutic biological agent or salt thereof; the particles having less than about 10% aggregates of the therapeutic biological agent; and the concentration of the therapeutic biological agent in the composition is from about 20 mg / mL to about 700 mg / mL.

[0140] In some embodiments, the disease or condition is cancer. In other embodiments, the disease or condition is an inflammatory disease or condition. In certain embodiments, the disease or condition is an immune disease. In certain other embodiments, the disease or condition is a kidney disease. In yet other embodiments, the disease or condition is a skin disease or condition. In yet other embodiments, the disease or condition is human immunodeficiency virus (HIV) infection.

[0141] Viscosity can play an important role in the handling and administration of injectable products.For suspension products, high viscosity can prevent the suspension from settling, but the drug product may be difficult to deliver through a 27-gauge needle because it requires a larger force to actuate the injection device, such as a syringe.Instead, using a wider needle or requiring a longer injection time can reduce patient compliance with treatment.

[0142] Particle size and dispersity also affect injectability, e.g., injectability. It is often recommended that particles be at least 3-10 times smaller than the inner diameter of the needle. Even a small fraction of the particle population larger than the inner diameter of the needle can clog the needle, resulting in the entire dose being wasted. As described herein, particles containing at least one therapeutic biological agent in suspension can achieve lower viscosities at higher concentrations. In some embodiments, particles as described herein can achieve greater than 90% protein loading while maintaining protein stability. The compositions described herein contain therapeutic biological agent concentrations of about 400 mg / mL at less than about 100 mPa·s, enabling subcutaneous delivery of therapeutic biological agents, e.g., proteins, mAbs, and drug compositions, without compromising their structural and functional bioactivity.

[0143] The particles described herein are, for example, a controlled distribution of discrete spheroids with characteristic sizes ranging from submicrometers to tens of micrometers, as opposed to the porous, monolithic "cake" typically produced during conventional freeze-drying. This morphology typically allows for a free-flowing powder (as described by a low Hausner ratio) without further processing.

[0144] In some embodiments, the particles are from about 0.1 to about 1000 μm, e.g., from about 1 to about 400 μm, from about 1 to about 200 μm, from about 1 to about 100 μm, from about 1 to about 50 μm, from about 1 to about 25 μm, from about 1 to about 10 μm, from about 10 to about 100 μm, from about 50 to about 100 μm, from about 50 to about 75 μm, or In other embodiments, the particles have a diameter of about 1 to about 100 μm, for example, about 4 to about 100 μm, about 10 to about 100 μm, or about 20 to about 50 μm.

[0145] In certain embodiments, the particles may contain one or more drugs, such as therapeutic biological agents. In other embodiments, the particles may have a diameter of about 0.1 to about 1000 μm, e.g., about 0.1 to about 90 μm, about 90 to about 230 μm, or about 0.1 to about 1 μm. In still other embodiments, the particles may have a size dispersity of about 0 to about 0.9, e.g., about 0 to about 0.7, about 0 to about 0.5, or about 0 to about 0.2. Methods for measuring particle size and particle distribution include imaging flow cytometry and image analysis of scanning electron micrographs of the particles, and the mean spherical radius or diameter can be calculated based on the cross-sectional area of ​​the particle projected onto the image plane.

[0146] In some embodiments, the sphericity of a particle can range from about 0.1 to about 1, e.g., at least about 0.2, about 0.4, about 0.6, or about 0.8. Methods for measuring particle sphericity include image analysis of scanning electron micrographs of the particle, where an average roundness is calculated based on the cross-sectional shape of the particle projected onto the image plane. Such roundness factors can be expanded to identify corresponding sphericities.

[0147] In other embodiments, the particles have less than about 20% aggregates or less than about 20% fragments of the therapeutic biological agent, e.g., less than about 19%, less than 18%, less than 17%, less than 16%, less than 15%, less than 14%, less than 13%, less than 12%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1.9%, less than 1.8%, less than 1.7%, less than 1.6%, less than 1.5%, less than 1.4%, less than 1.3%, less than 1.2%, less than 1.1%, less than 1%, less than 0.9%, less than 0.8%, less than 0.7%, less than 0.6%, less than 0.5%, less than 0.4%, less than 0.3%, less than 0.2%, or less than 0.1%. In some embodiments, the particles have less than about 10% aggregates or less than about 10% fragments of the therapeutic biological agent, e.g., less than about 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%. In certain embodiments, the particles have between about 3% and about 1% aggregates of the therapeutic biological agent. In certain other embodiments, the particles have between about 1% and about 0.5% aggregates of the therapeutic biological agent. In preferred embodiments, the particles are substantially free of aggregates of the therapeutic biological agent. In yet other embodiments, the particles are substantially free of fragments of the therapeutic biological agent. A suitable method for measuring aggregates and fragments of the biological agent can be achieved by using size exclusion chromatography (SEC).

[0148] In some embodiments, the particle formation process achieves less than a 50% change in charge variants in a population of therapeutic biological agents, e.g., antibodies or antibody fragments, compared to the therapeutic biological agents prior to particle formation (e.g., less than about 40%, 30%, 20%, 10%, 8%, 5%, 4%, 3%, or 1%). Charge variants can be acidic, basic, or neutral, and variations can be caused by post-translational modifications at terminal amino acids, such as asparagine deamidation or lysine glycosylation. For example, charge variants include loss of a positive charge due to loss of a C-terminal lysine residue, covalent attachment of the amine moieties of two lysine residues with a reducing sugar, or conversion of an N-terminal amine to a neutral amide via cyclization of an N-terminal glutamine. Negative charges on proteins, e.g., antibodies, can appear due to conversion of an asparagine residue to an aspartic acid and / or isoaspartic acid residue via a deamidation reaction.

[0149] An exemplary method for measuring charge variants involves cation exchange chromatography (CIEX), in which variants are quantified by dividing the area under the peak corresponding to a variant, e.g., an acidic, basic, or neutral population, by the cumulative area contained under all peaks in the sample spectrum. The change in percentage of a charge variant population between two samples, e.g., sample A and sample B, is calculated as the numerical difference in the percentage of each population variant, i.e., by subtracting the percentage of a particular variant, e.g., acidic, in sample B from the percentage of a particular variant, e.g., acidic, in sample A, or vice versa. In certain embodiments, the analysis can be similarly extended to all variants within a population.

[0150] In certain embodiments, the particles have less than about 50%, e.g., less than about 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% change in charge variants of the therapeutic biological agent compared to the starting therapeutic biological agent before particle formation. In preferred embodiments, the particles are substantially free of any change in charge variants of the therapeutic biological agent compared to the starting therapeutic biological agent before particle formation. A suitable method for measuring the change in charge variants of a therapeutic biological agent can be achieved by using cation exchange chromatography (CIEX).

[0151] Particles according to the present disclosure are circular. Circularity can serve as an indicator of particle shape. The particles described herein can have a characteristic circularity, e.g., a relative shape that is substantially circular. This characteristic describes and defines the particle's morphology based on its circularity. When a particle has a perfectly circular structure, the circularity is 1.0. Particles as described herein can have a circularity of about 0.8, 0.85, 0.9, 0.95, 0.96, 0.97, 0.98, or about 0.99. The diameter and circularity of a particle can be determined by image processing of an image observed under an electron microscope or a flow particle image analyzer. Circularity can also be determined by subjecting particles to circularity measurement and averaging the values ​​obtained. For example, circularity (circ) can be calculated using the following formula:

number

[0152] The term "perimeter" as used herein refers to the boundary line 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 a particle. The circularity of a particle can also be described as the ratio of the particle's smallest diameter to its largest diameter. For a perfect circle, this ratio is 1. The circularity percentage 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 handle images, such as images obtained by microscopy, in particular scanning electron microscopy (SEM) or transmission electron microscopy (TEM). In some embodiments, the particles have a circularity of about 0.10 to about 1.00, e.g., 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. In other embodiments, the particles have a circularity of about 0.80 to about 1.00. In still other embodiments, the particles have a circularity of about 0.85 to about 1.00. In certain other embodiments, the particles have a circularity of about 0.90 to about 1.00. In certain embodiments, the particles have a circularity of about 0.95 to about 1.00. In certain preferred embodiments, the particles have a circularity of about 0.98 to about 1.00. In preferred embodiments, the particles have a circularity of about 1.00. In some embodiments, a method for measuring the circularity of a particle involves image analysis of a scanning electron micrograph of the particle, and an average roundness is calculated based on the cross-sectional shape of the particle projected onto the image plane. Such roundness factors can be expanded to identify corresponding circularities.

[0153] In other embodiments, the residual moisture and / or solvent content of the dry components is less than about 7% by weight, e.g., less than about 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 weight. In some embodiments, the particles have less than about 7% by weight residual moisture. In still other embodiments, the particles have less than about 5% by weight residual moisture. In certain embodiments, the particles have less than about 3% by weight residual moisture. In preferred embodiments, the particles have less than about 1% by weight residual moisture.

[0154] In some embodiments, the particles have about 7% to about 1% residual moisture by weight. In other embodiments, the particles have about 5% to about 1% residual moisture by weight. In preferred embodiments, the particles have about 3% to about 1% residual moisture by weight. In certain preferred embodiments, the particles are substantially free of residual moisture by weight.

[0155] Exemplary methods for measuring moisture content include chemical titration methods, such as Karl Fischer titration, which involves an oven. Various solvents, including water, can also be measured using weight loss methods, which involve thermal excitation. Exemplary methods include thermogravimetric analysis with infrared spectroscopy (TGA-IR) or gas chromatography-flame ionization detector-gas chromatography (GC-FID / MS).

[0156] In some embodiments, the particles have less than about 50% internal void space, such as less than about 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1%. In certain embodiments, the particles may contain less than 25% internal void space, less than 10%, less than 5%, less than 3%, less than 1%, or less than 0.1% internal void space. In preferred embodiments, the particles are substantially free of any internal void space. Suitable methods for determining internal void space can be achieved by using a focused ion beam scanning electron microscope (FIB-SEM), which can be used to visualize "accessible" and "inaccessible" void space, or gas displacement pycnometry (Micromeritics Instrument Corporation, Norcross, Ga.), which can determine "accessible" void space (void space accessible from the surface rather than similar to a core-shell structure, which is "inaccessible from the surface"). Gas pycnometry is a common analytical technique that uses gas displacement methods to measure volume. An inert gas, such as helium or nitrogen, is used as the displacement medium. The true volume is the total volume minus the volume accessible by the gas. Density is calculated by dividing the sample weight by the true volume. The sample is sealed in an instrument compartment of known volume, receives the appropriate inert gas, and then expanded into another high-precision internal volume. The pressure before and after expansion is measured and used to calculate the sample volume. Dividing the sample weight by this volume gives the gas displacement density. Cross sections of exemplary particles of the present disclosure exhibit an absence of pores (substantially no internal void space) and low particle porosity, about 0.0005 g / cm, as shown by FIB-SEM or by gas pycnometry using helium at a temperature of about 22°C. 3 in Typically averaging about 1.3 g / cm with a standard deviation of 3 Achieve a density of

[0157] In other embodiments, the particles have a density of about 1 to about 6 g / cm 3 , for example, about 1 to about 5 g / cm 3 , about 1~3g / cm 3 , about 1~2g / cm 3 , about 1~1.5g / cm 3 or about 1.1 to about 1 .4g / cm 3 An exemplary method for measuring skeletal density is gas displacement pycnometry. In certain embodiments, the particles have a density of about 0.1 to about 5 g / cm 3 , for example, about 0.1 to about 2 .5g / cm 3 , about 0.1 to about 1.4 g / cm 3 , about 0.5 to about 1.4 g / cm 3 or about 1 0.0 to approx. 1.4g / cm 3 In certain embodiments, the particles have a density of about 1000 mg / mL ~ approx. 1500mg / mL, approx. 1050mg / mL ~ approx. 1500mg / mL, approx. 1100mg / mL ~ approx. 1500mg / mL, approx. 1150mg / mL ~ approx. 1500mg / mL, about 1200mg / mL to about 1500mg / mL, about 1250mg / mL to about 1500mg / mL, about 1300mg / mL to about 1500mg / mL, about 1310mg / mL to about 1500mg / mL, about 1320mg / mL to about 1500mg / mL, about 1330mg / mL to about 1500mg / mL, about 1340mg / mL ~ approx. 1500mg / mL, approx. 1350mg / mL ~ approx. 1500mg / mL, approx. 1360mg / mL ~ approx. 1500mg / mL, approx. 1370mg 1400mg / mL to about 1500mg / mL, about 1410mg / mL to about 1500mg / mL, about 1420mg / mL to about 1500mg / mL, about 1430mg / mL to about 1500mg / mL, about 1440mg / mL to about 1500mg / mL, about 1450mg / mL to about 1500mg / mL, about 1460mg / mL to about 1500mg / mL, about 1470mg / mL to about 1500mg / mL, about 1480mg / mL to about 1500mg / mL, or about 1490mg / mL to about 1500mg / mL. In preferred embodiments, the particles have a density of about 1000 mg / mL to about 1500 mg / mL, about 1300 mg / mL to about 1500 mg / mL, or about 1320 mg / mL to about 1500 mg / mL.

[0158] In some embodiments, the particles have greater than about 60% by weight of a therapeutic biological agent, e.g., greater than about 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% by weight of a therapeutic biological agent.

[0159] In some embodiments, the particles further comprise a carbohydrate, a pH adjuster, a salt, a chelating agent, a mineral, a polymer, a surfactant, a protein stabilizer, an emulsifier, a preservative, an amino acid, an antioxidant, a protein, an organic solvent, a paraben, a bactericide, a fungicide, a vitamin, a preservative, a nutrient medium, or a combination thereof. In certain embodiments, the particles further comprise a carbohydrate, a pH adjuster, a salt, a surfactant, a protein stabilizer, an emulsifier, an amino acid, or a combination thereof.

[0160] In other embodiments, the carbohydrate may be from the monosaccharide, disaccharide, oligosaccharide, or polysaccharide family. In some embodiments, the carbohydrate is dextran, trehalose, sucrose, agarose, mannitol, lactose, sorbitol, maltose, starch, alginate, xanthan, galactomannan, agar, agarose, or a combination thereof. In certain embodiments, the carbohydrate is dextran, trehalose, sucrose, agarose, mannitol, lactose, sorbitol, maltose, or a combination thereof. In preferred embodiments, the carbohydrate is trehalose, cyclodextrin, hydroxypropyl beta-cyclodextrin, sulfobutyl ether beta-cyclodextrin, or a combination thereof. Cyclodextrins are available in three different forms, α, β, and γ, based on the number of glucose monomers. The number of glucose monomers in α, β, and γ cyclodextrins can be 6, 7, or 8, respectively.

[0161] In some embodiments, the pH adjuster is acetate, citrate, glutamate, glycinate, histidine, lactate, maleate, phosphate, succinate, tartrate, bicarbonate, aluminum hydroxide, phosphoric acid, hydrochloric acid, DL-lactic acid / glycolic acid, phosphorylethanolamine, tromethamine, imidazole, glycylglycine, monosodium glutamate, sodium hydroxide, potassium hydroxide, or a combination thereof. In other embodiments, the pH adjuster is citrate, histidine, phosphate, succinate, sodium hydroxide, potassium hydroxide, or a combination thereof. In certain embodiments, the pH adjuster is hydrochloric acid or citric acid.

[0162] 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 a preferred embodiment, the salt is sodium chloride.

[0163] In some embodiments, the chelating agent is edetate disodium, 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 propylene glycol, glucose star polymer, silicone polymer, polydimethylsiloxane, polyethylene glycol, carboxymethylcellulose, poly(glycolic acid), poly(lactic-co-glycolic acid), polylactic acid, polycaprolactone (PCL), polyvinylpyrrolidone (PVP), ficoll, dextran, or a combination thereof.

[0164] 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, Aerosol-OT (sodium bis(2-ethylhexyl) sulfosuccinate), sodium laureth sulfate, lecithin, sorbitan esters, or combinations thereof. In some embodiments, surfactants include, but are 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, perfluorooctane sulfonate, alkyl ether phosphates; (iii) nonionic surfactants, such as alkylphenol ethoxylates (Triton X-100), fatty alcohol ethoxylates (octaethylene glycol monododecyl ether, cocamide diethanolamine, poloxamer, glycerol monostearate, fatty acid esters of sorbitol (monosodium phosphate), methylparaben, ... and (iv) zwitterionic surfactants, such as cocamidopropyl hydroxysultaine and 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS). In certain embodiments, the surfactant is a polysorbate, docusate, lecithin, or sorbitan ester. In preferred embodiments, the surfactant is a sorbitan ester, polysorbate 20, polysorbate 60, or polysorbate 80. In certain preferred embodiments, the surfactant is a sorbitan ester, polysorbate 20, or polysorbate 80. In certain other embodiments, the fatty acid ester of sorbitol is a sorbitan ester, such as span 20, span 40, span 60, span 80, or span 85.

[0165] In some embodiments, the protein stabilizer is acetyltryptophanate, caprylate, N-acetyltryptophan, trehalose, polyethylene glycol (PEG), poloxamer, polyvinylpyrrolidone, polyacrylic acid, poly(vinyl) polymer, poly In certain embodiments, the protein stabilizer is trehalose, polyethylene glycol (PEG), poloxamer, polyvinylpyrrolidone, polyacrylic acid, poly(vinyl) polymer, polyester, polyaldehyde, tertiary polymer, polyamino acid, hydroxyethyl starch, N-methyl-2-pyrrolidone, sorbitol, sucrose, mannitol, cyclodextrin, sugar, hydroxypropyl beta-cyclodextrin, sulfobutyl ether beta-cyclodextrin, or combinations thereof. In preferred embodiments, the protein stabilizer is trehalose, cyclodextrin, hydroxypropyl beta-cyclodextrin, sulfobutyl ether beta-cyclodextrin, or combinations thereof. As used herein, the stabilizer, which is used synonymously with the term "protein stabilizer" or "stabilizer," can be a salt, carbohydrate, sugar, or amino acid, preferably a carbohydrate or sugar, approved by an authority as a suitable additive or excipient in pharmaceutical compositions. In a preferred embodiment, the PEG is PEG200, PEG300, PEG3350, PEG8000, PEG10000, PEG20000, or a combination thereof.

[0166] Examples of emulsifiers suitable for use in the particles include, but are not limited to, lipophilic drugs having an HLB of less than 7, 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, glycerol monostearate, glyceryl distearate, glyceryl monopalmitate, and glyceryl dipalmitate; glyceryl-lactoesters of fatty acids; propylene glycol esters including propylene glycol monopalmitate, propylene glycol monostearate, and propylene glycol monooleate; sorbitan monostearate, sorbitan sesquioleate. sorbitan esters; fatty acids including stearic acid, palmitic acid, and oleic acid and soaps thereof; and mixtures thereof such as glyceryl monooleate, glyceryl dioleate, glyceryl monostearate, glyceryl distearate, glyceryl monopalmitate, and glyceryl dipalmitate; glyceryl-lactoesters of fatty acids; propylene glycol esters including propylene glycol monopalmitate, propylene glycol monostearate, and propylene glycol monooleate; sorbitan esters including sorbitan monostearate, sorbitan sesquioleate; fatty acids including stearic acid, palmitic acid, and oleic acid and soaps thereof; or combinations thereof. In some embodiments, the emulsifier is polysorbate 80, polysorbate 60, polysorbate 20, sorbitan monooleate, ethanolamine, polyoxyl 35 castor oil, polyoxyl 40 hydrogenated castor oil, carbomer 1342, corn oil mono-di-triglyceride, polyoxyethylated oleic acid glyceride, poloxamer, or a combination thereof. In preferred embodiments, the fatty acid ester of sorbitol is a sorbitan ester, such as span 20, span 40, span 60, span 80, span 85, etc. In certain preferred embodiments, the emulsifier is polysorbate 80, sorbitan monooleate, or a combination thereof.

[0167] In other embodiments, the preservative is phenol, m-cresol, benzyl alcohol, 2-phenyloxyethanol, chlorobutanol, neomycin, benzethonium chloride, glutaraldehyde, beta-propiolactone, or a combination thereof.

[0168] In certain embodiments, the amino acid is alanine, aspartic acid, cysteine, isoleucine, glutamic acid, leucine, methionine, phenylalanine, pyrrolidine, serine, selenocysteine, threonine, tryptophan, tyrosine, valine, asparagine, arginine, histidine, glycine, glutamine, proline, or a combination thereof. In a preferred embodiment, the amino acid is arginine, histidine, proline, asparagine, or a combination thereof.

[0169] In some embodiments, the antioxidant is glutathione, ascorbic acid, cysteine, N-acetyl-L-tryptophanate, tocopherol, histidine, methionine, or a combination thereof. 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. In still other embodiments, the preservative is methyl hydroxybenzoate, thimerosal, paraben, formaldehyde, castor oil, or a combination thereof. The paraben may be parahydroxybenzoate. In some embodiments, the disinfectant is benzalkonium chloride (a cationic surfactant), hypochlorite, peroxide, alcohol, a phenolic compound (e.g., carbolic acid), or a combination thereof.

[0170] In other embodiments, the fungicide is acibenzolar, 2-phenylphenol, allilazine, carvone, natamycin, potassium azide, or a combination thereof. In certain embodiments, the vitamin is thiamine, riboflavin, niacin, pantothenic acid, biotin, vitamin B6, vitamin B12, folate, niacin, ascorbic acid, calciferol, retinol, quinone, or a combination thereof. In still other embodiments, the preservative is sodium nitrate, sulfur dioxide, potassium sorbate, sodium sorbate, sodium benzoate, benzoic acid, methyl hydroxybenzoate, thimerosal, parabens, formaldehyde, castor oil, or a combination thereof.

[0171] Several nutrient media, preferably serum-free, alone or in combination, may be used in the present disclosure, including commercially available media or other media known in the art. Examples of such media (all serum-free or serum-depleted) include ADC-1, LPM (without bovine serum albumin), F10 (HAM), F12 (HAM), DCCM1, DCCM2, RPMI 1640, BGJ medium (Fitton-Jackson modified), Basal Medium Eagle (BME supplemented with Earl's salt base), Dulbecco's Modified Eagle Medium (DMEM, serum-free), Glasgow's Modified Eagle Medium (GMEM), Leibovitz's L-15 medium, McCoy's 5A medium, Medium M199 (M199E - with Earl's salt base), Medium M199 (M199H - with Hank's salt base), Minimum Essential Medium Eagle (MEM-E - with Earl'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 many others. Additionally, although serum-containing nutrient media may also be used in compositions according to the present disclosure, the use of serum-containing media is less preferred due to the possibility that serum may be contaminated with microbial agents and because patients may develop immune responses to certain antigenic components contained in the serum.

[0172] A plurality of particles comprising at least one therapeutic biological agent as described herein can be prepared by many methods and any method of forming particles disclosed in, for example, International Application Nos. PCT / US2017 / 063150, PCT / US2018 / 043774, PCT / US2019 / 033875, PCT / US20 / 15957, US62 / 799,696, US62 / 899,907, US62 / 899,981, US62 / 978,641, and US63 / 011,820, each of which is incorporated herein by reference in its entirety.

[0173] The particles of the present disclosure can be dispersed in an aqueous liquid carrier, a non-aqueous liquid carrier, such as an organic liquid, an ionic liquid carrier, or the like. The composition may be suspended in a solvent, a gel carrier, or a combination thereof to form a suspension composition. The vehicle for the suspension may further comprise, for example, carbohydrates, pH adjusters, salts, chelating agents, minerals, polymers, surfactants, amino acids, oligopeptides, biological excipients, chemical excipients, preservatives, antioxidants, parabens, bactericides, fungicides, vitamins, preservatives, analgesics, and / or nutrient media. In some embodiments, each of the other components may independently comprise 0.0001 to 99% (w / v) of the vehicle, e.g., 0.0001 to 90% (w / v), 0.0001 to 50% (w / v), 0.0001 to 10% (w / v), 0.0001 to 1% (w / v), or 0.0001 to 0.1% (w / v).

[0174] For aqueous or non-aqueous suspension compositions or formulations, high concentration trehalose solutions can be used to stabilize particles in suspension and prevent premature dissolution. In certain cases, sugars act as steric stabilizers when adsorbed onto particle surfaces, but can also act as "crowder" molecules when non-adsorbed. Crowder molecules can function by enhancing depletion repulsion. This stabilizing effect has also been described for other crowding agents, such as (i) polymers, such as PEG200, PEG300, PEG3350, PEG8000, PEG10000, PEG20000, poloxamers, polyvinylpyrrolidone, polyacrylic acid, poly(vinyl) polymers, polyesters, polyaldehydes, tertiary polymers, polyamino acids, or hydroxyethyl starch (note that these can be used alone or in combination); (ii) organic molecules, such as N-methyl-2-pyrrolidone (Miller et al. J. Pharm. Sci., 2012, 101, 3763-3778), and (iii) sugars or sugar alcohols, such as sorbitol, sucrose, and mannitol, among others. Other "crowding agents" include salts, such as ammonium sulfate, which can compete for water in hydration, and water-soluble organic liquids, such as N-methylpyrrolidone (NMP), which can reduce the solvent's dielectric constant and create an excluded volume effect.

[0175] In some embodiments, surfactants in the liquid carrier (either aqueous or non-aqueous) can act as charge stabilizers. The surfactants adsorb onto the surface of the particles and control the electrostatic interactions between them. The repulsive electrostatic forces generated by the addition of surfactants to the composition are sufficient to prevent significant aggregation of the particles in certain embodiments. The surfactants can also prevent adhesion to containers or vessels. In other embodiments, polymers can be added to the liquid carrier to act as steric stabilizers.

[0176] The term "viscosity" is used to describe the property of a fluid that acts to resist shear flow. For purposes of this disclosure, viscosity can be determined at 25°C using a rheometer fitted with a cone and plate (2° / 40 mm) at a specific shear rate, such as an AR-G2 rheometer (TA Instruments, USA). In certain embodiments, viscosity is measured at a shear rate in the Newtonian region. In other embodiments, viscosity is measured at 100 s -1 or higher shear rates, e.g., 1000 s -1 , or 1000s -1 Over 10,000s -1 Over or 50,000s -1 The term "low viscosity," as used herein, describes a composition, e.g., a liquid carrier, having a viscosity of less than about 200 mPa·s.

[0177] In some embodiments, the composition is 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 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 The viscosity of the composition is 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 (1 milliPascal second). In other embodiments, the composition has a viscosity of about 0.01 mPa·s to about 10,000 mPa·s, e.g., about 0.01 mPa·s to about 1,000 mPa·s, about 0.01 mPa·s to about 100 mPa·s, about 0.01 mPa·s to about 50 mPa·s, about 0.01 mPa·s to about 25 mPa·s, about 0.01 mPa·s to about 10 mPa·s, about 0.01 mPa·s to about 5 mPa·s, or about 0.01 mPa·s to about 1 mPa·s. In certain embodiments, the viscosity of the composition can be in the range of about 0.27 mPa·s to about 200 mPa·s, e.g., about 0.27 mPa·s to about 50 mPa·s, about 1 mPa·s to about 30 mPa·s, or about 20 mPa·s to about 50 mPa·s. In still other embodiments, the viscosity of the composition ranges from about 0.27 mPa·s to about 200 mPa·s, e.g., from about 0.27 mPa·s to about 100 mPa·s, from about 0.27 mPa·s to about 50 mPa·s, from about 0.27 mPa·s to about 30 mPa·s, from about 1 mPa·s to about 20 mPa·s, or from about 1 mPa·s to about 15 mPa·s. In certain embodiments, the viscosity is measured at shear rates in the Newtonian region. In other embodiments, the viscosity is measured at shear rates of less than or greater than about 100 s-1, e.g., at or above about 1000 s-1. The composition may comprise from about 5 to about 90% particles by volume, e.g., from about 20 to about 90%, from about 40 to about 80%, from about 50 to about 60%, or from about 70 to about 90%.The composition may have a concentration of therapeutic biological agent of about 0.0001 to about 1000 mg / mL, e.g., about 100 to about 900, about 150 to about 800, or about 200 to about 700 mg / mL. Methods for controlling viscosity include temperature control and viscosity-modifying additives. Liquid mixtures may also be used to control viscosity. The units "mPa·s" and "cP" are used interchangeably herein in the broadest sense.

[0178] In other embodiments, the composition has a viscosity of less than about 200 mPa·s. In some embodiments, the composition has a viscosity of less than about 150 mPa·s. In certain embodiments, the composition has a viscosity of less than about 100 mPa·s. In still other embodiments, the composition has a viscosity of less than about 80 mPa·s. In certain other embodiments, the composition has a viscosity of less than about 50 mPa·s. In some embodiments, the composition has a viscosity of less than about 40 mPa·s. In other embodiments, the composition has a viscosity of less than about 30 mPa·s. In certain embodiments, the composition has a viscosity of less than about 20 mPa·s. In still other embodiments, the composition has a viscosity of less than about 10 mPa·s. In certain other embodiments, the composition has a viscosity of less than about 5 mPa·s. In some embodiments, the composition has a viscosity of less than about 3 mPa·s. In other embodiments, the composition has a viscosity of less than about 2.5 mPa·s.

[0179] Certain embodiments of the present disclosure described herein allow for high concentrations of therapeutic biological agents in particles and high concentrations of particles in liquid carriers, which in some embodiments may be achieved by mixing particles of various sizes.

[0180] In some embodiments, the treatment methods described herein comprise administering a steroid hormone to a subject in need thereof, comprising administering to the subject a steroid hormone from about 20 mg / mL to about 700 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, 650, 675 mg / mL to about 700 mg / mL; from about 20 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, 650, 675 mg / mL to about 700 mg / mL;0, 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 600 mg / mL, for example, 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 60 0 mg / mL; about 20 mg / mL to about 575 mg / mL, for example, 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, for example, about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 50 0, 525 mg / mL to about 550 mg / mL; about 20 mg / mL to about 525 mg / mL, for example, 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, for example, 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, for example, 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, for example, 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, for example, 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, for example, 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, for example, about 20, 30, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350 mg / mL to about 375 mg 25, 26, 27, 28, 29, 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, 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, is used. In other embodiments, the concentration of the therapeutic biological agent in the composition is from about 20 mg / mL to about 650 mg / mL. In certain embodiments, the concentration of the therapeutic biological agent in the composition is from about 100 mg / mL to about 500 mg / mL. In still other embodiments, the concentration of the therapeutic biological agent in the composition is from about 200 mg / mL to about 400 mg / mL. In preferred embodiments, the concentration of the therapeutic biological agent in the composition is from about 300 mg / mL to about 400 mg / mL. In certain preferred embodiments, the concentration of the therapeutic biological agent in the composition is from about 350 mg / mL to about 400 mg / mL. In some embodiments, the concentration of the therapeutic biological agent in the composition is from about 300 mg / mL to about 500 mg / mL.

[0181] In certain embodiments, insoluble particulate matter having a characteristic size of about 100 μm or greater that persists upon dissolution in an aqueous liquid is referred to as visible particles (VP). In preferred embodiments of the present disclosure described herein, the compositions are substantially free of visible particles (VP). Certain In preferred embodiments, the aqueous liquid is water, an aqueous buffer solution, or a physiologically suitable aqueous liquid. As used herein, the term "physiologically suitable aqueous liquid" refers to any water-containing body fluid distributed in the extracellular compartment, such as extracellular fluid, interstitial fluid, intravascular fluid (blood, plasma, and lymph), and cerebrospinal fluid.

[0182] In another embodiment, insoluble particulate matter having a characteristic size of about 1 μm to about 100 μm that persists upon dissolution in an aqueous liquid is referred to as subvisible particles (SvP). SvP are present in an amount of about 0 to 100,000,000 per mL, e.g., about 0 to about 10,000,000 per mL, about 0 to about 1,000,000 per mL, about 0 to about 500,000 per mL, about 0 to about 100,000 per mL, about 0 to about 50,000 per mL, about 0 to about 10,000 per mL, about 0 to about 6,000 per mL, about 0 to about 1,000 per mL, about 0 to about 600 per mL, about 0 to about 250 per mL, about 0 to about 100 per mL, about 0 to about 60 per mL, or about 0 to about 10 per mL. In other embodiments, the number of particles having a characteristic size of 10 μm or greater is about 0 to about 6,000 per mL, e.g., about 0 to about 1,000 per mL, about 0 to about 100 per mL, about 0 to about 10 per mL, about 0 to about 5 per mL, about 0 to about 3 per mL, or about 0 to about 1 per mL. In certain embodiments, the number of particles having a characteristic size of 25 μm or greater is about 0 to about 600 per mL, e.g., about 0 to about 100 per mL, about 0 to about 10 per mL, about 0 to about 3 per mL, about 0 to about 1 per mL, about 0 to about 0.5 per mL, or about 0 to about 0.1 per mL. Exemplary methods for measuring SvP include analysis of the therapeutic biological agent by a Coulter Counter, HIAC Royco, or microfluidic imaging system after reconstitution and dilution of the therapeutic biological agent to a standard concentration, e.g., about 100 mg / mL or about 1 mg / mL. In still other embodiments, the composition, when dissolved in an aqueous liquid, has a concentration of insoluble sub-visible particles (SvP) of about 0 to about 100,000,000 per mL of particles greater than about 10 μm. In certain embodiments, the composition, when dissolved in an aqueous liquid, has a concentration of insoluble sub-visible particles (SvP) of about 0 to about 6000 per mL of particles greater than about 10 μm. In preferred embodiments, the composition, when dissolved in an aqueous liquid, has a concentration of insoluble sub-visible particles (SvP) of about 0 to about 600 per mL of particles greater than about 25 μm.In certain preferred embodiments, the composition, when dissolved in an aqueous liquid, is substantially free of insoluble subvisible particles (SvP).

[0183] In certain embodiments, the composition, when dissolved in an aqueous liquid, has a concentration of insoluble sub-visible particles (SvP) having a characteristic size of greater than about 10 μm of from about 0 particles per mL to about 100,000,000 particles per mL. In some embodiments, the composition, when dissolved in an aqueous liquid, has a concentration of insoluble sub-visible particles (SvP) having a characteristic size of greater than about 10 μm of from about 0 particles per mL to about 6000 particles per mL. In other embodiments, the composition, when dissolved in an aqueous liquid, has a concentration of insoluble sub-visible particles (SvP) having a characteristic size of greater than about 25 μm of from about 0 particles per mL to about 600 particles per mL. In preferred embodiments, the aqueous liquid is water, an aqueous buffer solution, or a physiologically suitable aqueous liquid.

[0184] In some embodiments, insoluble particulate matter having a characteristic size of about 100 nm to about 1 μm that persists upon dissolution in aqueous liquids is referred to as submicron particles (SMPs), sometimes known as nanoparticles. The presence of such SMPs is believed to contribute to immunogenicity and, as such, should be avoided to minimize such effects. Quantitatively, SMPs are present in a concentration of about 0-5×10 per mL. 12 , for example, about 0 to about 0.5 × 10 per mL 12 , approximately 0 to approximately 50 × 10 per mL 9 , approximately 0 to approximately 10 × 10 per mL 9 , approximately 0 to approximately 5 × 10 per mL 9 , approximately 0 to approximately 0.5 × 10 per mL 9 , approximately 0 to approximately 50 × 10 per mL 6 , approximately 0 to approximately 1 × 10 per mL 6 , about 0 to about 500,000 per 1 mL, about 0 to about 200,000 per 1 mL, about 0 to about 100,000 per 1 mL, The SMP is present in an amount of about 0 to about 10,000 per mL, about 0 to about 5000 per mL, or about 0 to about 1000 per mL. Exemplary methods for quantitatively measuring SMP include analysis of the therapeutic biological agent by NanoSight, a microfluidic imaging system, asymmetric field-flow fractionation coupled to multi-angle laser light scattering (AF4MALS), dynamic light scattering (DLS), or FlowCam imaging after reconstitution and dilution of the therapeutic biological agent to a standard concentration, e.g., about 100 mg / mL, about 1 mg / mL, or about 1 μg / mL. Qualitatively, the SMP is in a range equivalent to the starting monomer therapeutic biological agent solution. In preferred embodiments, the composition is substantially free of submicron particles (SMP) upon dissolution in an aqueous liquid. In certain preferred embodiments, the aqueous liquid is water, an aqueous buffer, or a physiologically suitable aqueous liquid.

[0185] As disclosed herein, the term "immunogenicity" refers to the elicitation of an immune response by an injected composition of a therapeutic biological agent (antigen), while "antigenicity" refers to the reaction of a composition of a therapeutic biological agent with pre-existing antibodies. Collectively, antigenicity and immunogenicity are referred to as "immunoreactivity." In preferred embodiments, the composition has substantially similar immunogenicity compared to an aqueous composition comprising a therapeutic biological agent in monomeric form. In certain preferred embodiments, the composition is substantially non-immunogenic.

[0186] The ratio between toxic and therapeutic effect for a particular composition is its therapeutic index, LD 50 (the amount of compound that is lethal to 50% of the population) and ED 50 Therapeutic indices can be expressed as a ratio between the ED (the amount of compound effective in 50% of the population). Compositions that exhibit high therapeutic indices are preferred. Therapeutic index data obtained from cell culture assays and / or animal studies can be used in formulating a dosage range for use in humans. The dosage of such compounds preferably is within the ED range with little or no toxicity. 50The dosage may vary within this range depending on the dosage form employed and the route of administration utilized, e.g., subcutaneous injection. See Fingl et al. in THERAPEUTICS, Ch. 1, p. 1, 1975. The exact composition, route of administration and dosage can be chosen by the individual physician in view of the patient's condition and the particular method used to deliver the composition.

[0187] Oncology or anti-inflammatory drugs often have significant side effects due to off-target toxicity, such as hematotoxicity, neurotoxicity, cardiotoxicity, hepatotoxicity, nephrotoxicity, ototoxicity or cerebrotoxicity. For example, taxanes, such as docetaxel, can cause the following adverse effects: infection, neutropenia, anemia, febrile neutropenia, hypersensitivity, thrombocytopenia, myelotoxicity, myelosuppression, neuropathy, dysgeusia, dyspnea, constipation, anorexia, nail disorders, fluid retention, asthenia, pain, nausea, diarrhea, vomiting, fatigue, nonspecific neurocognitive problems, vertigo, encephalopathy, mucositis, alopecia, skin reactions, myalgia, or a combination thereof.

[0188] In some embodiments of the present disclosure described herein, the toxicity is hematotoxicity, neurotoxicity, gastrointestinal toxicity, cardiotoxicity, hepatotoxicity, nephrotoxicity, ototoxicity, or cerebrotoxicity. In other embodiments, the side effects associated with toxicity can be neutropenia, leukopenia, thrombocytopenia, myelotoxicity, myelosuppression, neuropathy, fatigue, nonspecific neurocognitive problems, vertigo, encephalopathy, anemia, dysgeusia, dyspnea, constipation, anorexia, nail disorders, fluid retention, asthenia, pain, nausea, vomiting, mucositis, hair loss, skin reactions, myalgia, hypersensitivity, anaphylaxis, or a combination thereof.

[0189] In certain embodiments, the composition has substantially similar toxicity compared to aqueous compositions comprising a monomeric therapeutic biological agent. In preferred embodiments, the composition has reduced toxicity compared to aqueous compositions comprising a monomeric form of the therapeutic biological agent. In certain preferred embodiments, the composition is substantially non-toxic.

[0190] The present disclosure as described herein relates to highly concentrated compositions comprising a plurality of particles suspended in a low-viscosity pharmaceutically acceptable liquid carrier, each particle comprising at least one therapeutic biological agent, wherein the composition, upon dissolution in water, buffer, or other physiologically suitable aqueous liquid, e.g., a biological fluid in a patient's body, has substantially similar turbidity compared to a similar aqueous composition comprising a monomeric therapeutic biological agent. The term "turbidity" refers to the cloudiness or haze of the fluid caused by individual particles that remain insoluble after dissolution in water, buffer, or other physiologically suitable aqueous liquid, e.g., a biological fluid in a patient's body, at a desired concentration. As used herein, "physiologically suitable" conditions such as those encountered in a mammal or human can be applied. One of skill in the art can determine the most appropriate set of conditions to test according to the end use of the compositions described herein. In some embodiments, upon dissolution in an aqueous liquid, the composition has substantially similar turbidity compared to an aqueous composition comprising a monomeric therapeutic biological agent. In preferred embodiments, upon dissolution in an aqueous liquid, the composition is substantially free of turbidity. In certain preferred embodiments, the aqueous liquid is water, an aqueous buffer solution, or a physiologically suitable aqueous liquid.

[0191] In certain embodiments, the present disclosure relates to a highly concentrated composition with low turbidity comprising a plurality of particles comprising a carbohydrate, a pH adjuster, a salt, a surfactant, a protein stabilizer, an emulsifier, an amino acid, and a therapeutic biological agent in a non-aqueous liquid carrier. In a preferred embodiment, the present disclosure relates to a highly concentrated composition with low turbidity comprising a plurality of particles comprising trehalose, arginine hydrochloride, sodium succinate, succinic acid, citric acid, sodium citrate, histidine, histidine hydrochloride, sodium chloride, hydroxypropyl beta-cyclodextrin, sulfobutyl ether beta-cyclodextrin, polysorbate or sorbitan monooleate, and an antibody in ethyl oleate. In certain preferred embodiments, the composition is substantially free of turbidity upon dissolution in water, an aqueous buffer, or any physiologically suitable aqueous liquid.

[0192] In certain embodiments, the particles can be suspended in a non-aqueous or aqueous liquid carrier, thereby forming a non-aqueous or aqueous pharmaceutically acceptable composition. Importantly, the process of producing a non-aqueous or aqueous composition having at least one therapeutic biological agent does not significantly alter the structure or biological activity of the biological agent as described herein. Furthermore, in certain other embodiments, the present disclosure allows for the delivery of higher doses of therapeutic biological agents while minimizing delivery volume, shortening administration time, and / or reducing pain.

[0193] In some embodiments, the liquid carrier is non-aqueous or aqueous. In other embodiments, the liquid carrier is non-aqueous. In yet other embodiments, the liquid carrier is aqueous.

[0194] In other embodiments, the non-aqueous liquid carrier is an organic solvent or an ionic liquid. In some embodiments, the organic solvent is selected from the group consisting of benzyl benzoate, coconut oil, cottonseed oil, fish oil, grapeseed oil, hazelnut oil, hydrogenated vegetable oil, olive oil, palm kernel oil, peanut oil, peppermint oil, safflower oil, sesame oil, soybean oil, sunflower oil, walnut oil, corn oil, acetone, ethyl acetate, ethyl lactate, dimethylacetamide, dimethyl isosorbide, dimethyl sulfoxide, glycofurol, diglyme, methyl tert-butyl ether, N-methylpyrrolidone, perfluorodecalin, polyethylene glycol, 2-pyrrolidone, tetrahydrofurfuryl alcohol, triglycerides, methyl methyl ether, methyl tert-butyl ether, methyl methyl pyr ... methyl methyl ether, methyl tert-butyl ether, methyl methyl pyrrolidone, perfluorodecalin, polyethylene glycol, 2-pyrrolidone, tetrahydrofurfuryl alcohol, methyl methyl ether, methyl tert-butyl ether, methyl methyl ether, methyl tert-butyl ether, methyl methyl ether, methyl tert-butyl ether, methyl tert-butyl ether, methyl tert-butyl ether, methyl tert-butyl ether, methyl tert-butyl ether, methyl tert-butyl ether, methyl tert-butyl ether, methyl tert-butyl ether, methyl tert-butyl ether, methyl tert- glycerides, e.g., Miglyol, fractionated vegetable fatty acid C8 and C10 triglycerides, propylene glycol diesters of saturated vegetable fatty acids C8 and C10, 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, polyoxyl 40 hydrogenated castor oil, polyoxyl 35 castor oil, simple alcohols, e.g., ethanol, octanol, hexanol, decanol, propanol and butanol, gamma-butyrolactone, eugenol, tyrolactone, tocopherol, octafluoropropane, (perfluorohexyl)octane, n-acetyltryptophan, ethyl laurate, methyl caprylate, methyl caprate, methyl myristate, methyl oleate, methyl linoleate, dimethyl adipate, dibutyl suberate, diethyl sebacate, ethyl macadamiate, trimethylolpropane triisostearate, isopropyl laurate, isopropyl myristate, diethyl succinate, polysorbate esters, ethanolamine, propanoic acid, citral, anisole, anethole, benzaldehyde, linalool, caprolactone, phenol, thioglycerol, dimethylacetamide, diethylene glycol monoethyl ether, propylene carbonate, solketal, isosorbide dimethyl ether, ethyl formate, ethylhexyl acetate, or combinations thereof. In yet other embodiments, the ionic liquid comprises pyridinium, pyridazinium, pyrimidinium, pyrazinium, imidazolium, pyrazolium, thiazolium, oxazolium, triazolium, ammonium, sulfonium, halide, sulfate, sulfonate, carbonate, phosphate, bicarbonate, nitrate, acetate, PF6-, BF4-, triflate, nonaflate, bis(trifyl)amide, trifluoroacetate, heptafluorobutanoate, haloaluminate, or a combination thereof. In preferred embodiments, the organic solvent is ethyl oleate, a triglyceride such as Miglyol, ethyl laurate, ethyl macadamiate, ethyl caprate, diethyl succinate, diethylene glycol monoethyl ether, propylene carbonate, or a combination thereof. In certain preferred embodiments, the organic solvent is ethyl oleate or a triglyceride.

[0195] In some embodiments, the aqueous liquid carrier is water, 0.9% saline, lactated Ringer's solution, dextrose 5%, or a buffer solution. In other embodiments, the buffer solution is acetate buffer, histidine buffer, succinate buffer, HEPES buffer, Tris buffer, carbonate buffer, citrate buffer, phosphate buffer, phosphate-buffered saline, glycine buffer, barbital buffer, cacodylate buffer, ammonium formate buffer, urea solution, or a combination thereof. In a preferred embodiment, the aqueous liquid carrier is water.

[0196] In certain embodiments, the non-aqueous or aqueous liquid carrier further comprises a carbohydrate, a pH adjuster, a salt, a chelating agent, a mineral, a polymer, a surfactant, a protein stabilizer, an emulsifier, a preservative, an amino acid, an antioxidant, a protein, an organic solvent, a paraben, a bactericide, a fungicide, a vitamin, a preservative, a nutrient medium, an analgesic, or a combination thereof. In preferred embodiments, the non-aqueous or aqueous liquid carrier further comprises a carbohydrate, a pH adjuster, a salt, a surfactant, a protein stabilizer, an emulsifier, an amino acid, or a combination thereof. In certain preferred embodiments, the aqueous liquid carrier further comprises a carbohydrate, a pH adjuster, a salt, a surfactant, a protein stabilizer, an emulsifier, an amino acid, or a combination thereof.

[0197] In other embodiments, the carbohydrate may be from the monosaccharide, disaccharide, oligosaccharide, or polysaccharide family. In some embodiments, the carbohydrate is dextran, trehalose, sucrose, agarose, mannitol, lactose, sorbitol, maltose, starch, alginate, xanthan, galactomannan, agar, agarose, or a combination thereof. In certain embodiments, the carbohydrate is dextran, trehalose, sucrose, agarose, mannitol, lactose, sorbitol, maltose, or a combination thereof. In preferred embodiments, the carbohydrate is trehalose, cyclodextrin, hydroxypropyl beta-cyclodextrin, sulfobutyl ether beta-cyclodextrin, or a combination thereof. Cyclodextrins are available in three different forms, α, β, and γ, based on the number of glucose monomers. The number of glucose monomers in α, β, and γ cyclodextrins can be 6, 7, or 8, respectively.

[0198] In some embodiments, the pH adjuster is acetate, citrate, glutamate, glycerin, or the like. The pH adjuster is selected from the group consisting of citrate, histidine, lactate, maleate, phosphate, succinate, tartrate, bicarbonate, aluminum hydroxide, phosphoric acid, hydrochloric acid, DL-lactic acid / glycolic acid, phosphorylethanolamine, tromethamine, imidazole, glycylglycine, monosodium glutamate, sodium hydroxide, potassium hydroxide, and combinations thereof. In other embodiments, the pH adjuster is citrate, histidine, phosphate, succinate, sodium hydroxide, potassium hydroxide, and combinations thereof. In certain embodiments, the pH adjuster is hydrochloric acid or citric acid.

[0199] 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 a preferred embodiment, the salt is sodium chloride.

[0200] In some embodiments, the chelating agent is edetate disodium, 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 propylene glycol, glucose star polymer, silicone polymer, polydimethylsiloxane, polyethylene glycol, carboxymethylcellulose, poly(glycolic acid), poly(lactic-co-glycolic acid), polylactic acid, polycaprolactone (PCL), polyvinylpyrrolidone (PVP), ficoll, dextran, or a combination thereof.

[0201] 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, Aerosol-OT (sodium bis(2-ethylhexyl) sulfosuccinate), sodium laureth sulfate, lecithin, sorbitan esters, or combinations thereof. In some embodiments, surfactants include, but are 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, perfluorooctane sulfonate, alkyl ether phosphates; (iii) nonionic surfactants, such as alkylphenol ethoxylates (Triton X-100), fatty alcohol ethoxylates (octyl methyl ammonium bromide), and the like. (iv) zwitterionic surfactants, such as cocamidopropyl hydroxysultaine and 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS). In certain embodiments, the surfactant is a polysorbate, docusate, lecithin, or sorbitan ester. In preferred embodiments, the surfactant is a sorbitan ester, polysorbate 20, or polysorbate 80.

[0202] In some embodiments, the protein stabilizer is acetyltryptophanate, caprylate, N-acetyltryptophan, trehalose, polyethylene glycol (PEG), poloxamer, polyvinylpyrrolidone, polyacrylic acid, poly(vinyl) polymer, polyester, polyaldehyde, tertiary polymer, polyamino acid, hydroxyethyl starch, N-methyl-2-pyrrolidone, sorbitol, sucrose, mannitol, sugars, or combinations thereof. The protein stabilizer is selected from the group consisting of acrylic acid, poly(vinyl) polymers, polyesters, polyaldehydes, tertiary polymers, polyamino acids, hydroxyethyl starch, N-methyl-2-pyrrolidone, sorbitol, sucrose, mannitol, cyclodextrin, hydroxypropyl beta-cyclodextrin, sulfobutyl ether beta-cyclodextrin, acetyltryptophanate, caprylate, N-acetyltryptophan, and combinations thereof. In preferred embodiments, the protein stabilizer is trehalose, polyethylene glycol (PEG), cyclodextrin, hydroxypropyl beta-cyclodextrin, sulfobutyl ether beta-cyclodextrin, and combinations thereof. In certain preferred embodiments, the PEG is PEG200, PEG300, PEG3350, PEG8000, PEG10000, PEG20000, and combinations thereof. Stabilizers, used synonymously with the term "stabilizer," may be salts, carbohydrates, sugars, or amino acids, preferably carbohydrates or sugars, recognized by authorities as suitable additives or excipients in pharmaceutical compositions, as described herein.

[0203] Examples of emulsifiers suitable for use in non-aqueous or aqueous liquid carriers include, but are not limited to, lipophilic agents having an HLB of less than 7, 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; glyceryl-lactoesters of fatty acids; propylene glycol esters including propylene glycol monopalmitate, propylene glycol monostearate, and propylene glycol monooleate; sorbitan monostearate, sorbitan sesquioleate. sorbitan esters including sorbitan monostearate, sorbitan sesquioleate; fatty acids including stearic acid, palmitic acid, and oleic acid and soaps thereof; and mixtures thereof such as glyceryl monooleate, glyceryl dioleate, glyceryl monostearate, glyceryl distearate, glyceryl monopalmitate, and glyceryl dipalmitate; glyceryl-lactoesters of fatty acids; propylene glycol esters including propylene glycol monopalmitate, propylene glycol monostearate, and propylene glycol monooleate; sorbitan esters including sorbitan monostearate, sorbitan sesquioleate; fatty acids including stearic acid, palmitic acid, and oleic acid and soaps thereof; or combinations thereof. In some embodiments, the emulsifier is polysorbate, sorbitan monooleate, ethanolamine, polyoxyl 35 castor oil, polyoxyl 40 hydrogenated castor oil, carbomer 1342, corn oil-mono-di-triglyceride, polyoxyethylated oleic acid glyceride, poloxamer, or a combination thereof. In a preferred embodiment, the emulsifier is polysorbate 80, sorbitan monooleate, or a combination thereof.

[0204] In other embodiments, the preservative is phenol, m-cresol, benzyl alcohol, 2-phenyloxyethanol, chlorobutanol, neomycin, benzethonium chloride, glutaraldehyde, beta-propiolactone, or a combination thereof.

[0205] In certain embodiments, the amino acid is alanine, aspartic acid, cysteine, isoleucine, glutamic acid, leucine, methionine, phenylalanine, pyrrolysine, serine, selenocysteine, threonine, tryptophan, tyrosine, valine, asparagine, arginine, histidine, glycine, glutamine, proline, or a combination thereof. In preferred embodiments, the amino acid is arginine, histidine, proline, asparagine, or a combination thereof.

[0206] In some embodiments, the antioxidant is glutathione, ascorbic acid, cysteine, N-acetyl-L-tryptophanate, tocopherol, histidine, methionine, or any of the foregoing. 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. In still other embodiments, the preservative is methyl hydroxybenzoate, thimerosal, paraben, formaldehyde, castor oil, or a combination thereof. The paraben can be parahydroxybenzoate. In some embodiments, the disinfectant is benzalkonium chloride (a cationic surfactant), hypochlorite, peroxide, alcohol, a phenolic compound such as carbolic acid, or a combination thereof.

[0207] In other embodiments, the fungicide is acibenzolar, 2-phenylphenol, allilazine, carvone, natamycin, potassium azide, or a combination thereof. In certain embodiments, the vitamin is thiamine, riboflavin, niacin, pantothenic acid, biotin, vitamin B6, vitamin B12, folic acid, niacin, ascorbic acid, calciferol, retinol, quinone, or a combination thereof. In still other embodiments, the preservative is sodium nitrate, sulfur dioxide, potassium sorbate, sodium sorbate, sodium benzoate, benzoic acid, methyl hydroxybenzoate, thimerosal, parabens, formaldehyde, castor oil, or a combination thereof.

[0208] Several nutrient media, preferably serum-free, alone or in combination, may be used in the present disclosure, including commercially available media or other media known in the art. Examples of such media (all serum-free or serum-depleted) include ADC-1, LPM (without bovine serum albumin), F10 (HAM), F12 (HAM), DCCM1, DCCM2, RPMI 1640, BGJ medium (Fitton-Jackson modified), Basal Medium Eagle (BME supplemented with Earl's salt base), Dulbecco's Modified Eagle Medium (DMEM, serum-free), Glasgow's Modified Eagle Medium (GMEM), Leibovitz's L-15 medium, McCoy's 5A medium, Medium M199 (M199E - with Earl's salt base), Medium M199 (M199H - with Hank's salt base), Minimum Essential Medium Eagle (MEM-E - with Earl'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 many others. Additionally, serum-containing nutrient media may also be used in compositions according to the present disclosure.

[0209] In some embodiments, the analgesic agent is paracetamol, a histamine receptor antagonist, such as an H1 or H2 blocker, an NSAID, a COX-2 inhibitor, celecoxib, rofecoxib, valdecoxib, parecoxib, lumiracoxib, etoricoxib, firocoxib, acetaminophen, an opiate, dextropropoxyphene, codeine, tramadol, anileridine, pethidine, hydrocodone, morphine, oxycodone, methadone, diacetylmorphine, hydromorphone, oxymorphone, levorphanol, buprenorphine, fentanyl, sufentanil, etorphine, carfentanil, dihydromorphine, dihydrocodeine, thebaine, papaverine, diproqualone, flupirtine, a tricyclic antidepressant, acetaminophen, or lidocaine, or a combination thereof. In certain embodiments, the analgesic agent is acetaminophen or lidocaine.

[0210] In preferred embodiments according to the present disclosure as described herein, the particle compositions have improved stability of the therapeutic biological agents compared to aqueous compositions containing the therapeutic biological agents in monomeric form. A "stable" composition is one in which all therapeutic biological agents therein essentially retain their physical and / or chemical stability and / or biological activity upon storage at the intended storage temperature, e.g., 4-40°C. It is desirable for the formulation to essentially retain its physical and chemical stability and its biological activity upon storage. The storage period is generally selected based on the intended shelf life of the composition. Furthermore, the composition The composition should be stable following freezing and thawing (e.g., to -70°C), e.g., following 1, 2, or 3 cycles of freezing and thawing. Various analytical techniques for measuring protein stability are available in the art, e.g., Peptide and Stability is reviewed in Protein Drug Delivery, 247-301, Vincent Lee Ed., Marcel Dekker, Inc., New York, NY, Pub. (1991) and Jones, A. Adv. Drug Delivery Rev. 10:29-90 (1993). Stability can be measured for a selected period of time at a selected temperature. Stability can be qualitatively and / or quantitatively assessed by a variety of different methods, including assessing aggregate formation (e.g., by measuring turbidity using size exclusion chromatography and / or by visual inspection); assessing charge heterogeneity using cation exchange chromatography or capillary zone electrophoresis; amino- or carboxy-terminal sequence analysis; mass spectrometry; SDS-PAGE analysis comparing reduced and intact antibodies; peptide map (e.g., trypsin or LYS-C) analysis; assessing antibody biological activity or antigen-binding function, and the like. In some embodiments, the composition is stable for at least one month. In yet other embodiments, the composition is stable for at least two months. In certain other embodiments, the composition is stable for at least 3 months. In certain preferred embodiments, the composition is stable at 40° C. for at least 3 months.

[0211] In certain embodiments, the composition further comprises at least one pharmaceutically acceptable additive, diluent, excipient, carrier, or combination thereof.

[0212] In some embodiments, the compositions and methods disclosed herein further comprise a pharmaceutically effective amount of at least one hyaluronan degrading agent, which can be administered simultaneously with the composition, sequentially, or intermittently. Subcutaneous tissue or extracellular matrix consists of a network of fibrous proteins embedded in a viscoelastic gel of glycosaminoglycans. Hyaluronan is the predominant glycosaminoglycan in subcutaneous tissue. Hyaluronan is secreted into the interstitium by fibroblasts as a viscous polymer, which is subsequently degraded locally in the lymphatics by the action of hyaluronidase. Glycosaminoglycans are complex linear polysaccharides of the extracellular matrix, characterized, as in the case of hyaluronan, by a repeating disaccharide structure of N-substituted hexosamine and uronic acid.

[0213] The hyaluronan degrading agents of the present disclosure can enhance subcutaneous administration of compositions comprising multiple particles, for example, by increasing and / or enhancing the volume of the composition administered by syringe injection, thereby improving the absorption of the therapeutic biological agent. The use of hyaluronan degrading agents, such as hyaluronidase, can improve the subcutaneous administration of therapeutic biological agents into the systemic circulation through the reversible hydrolysis 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 larger volumes to be safely and comfortably administered into the subcutaneous tissue. Furthermore, the degradation of hyaluronan temporarily reduces the viscosity of the subcutaneous tissue, facilitating the dispersion of injected fluids and their absorption. The effects of hyaluronidase are local and reversible, with complete reconstitution of the hyaluronan tissue occurring within 24 to 48 hours. For example, Frost, GI, “Recombinant human hyaluronidase (rHuPH20): an enabling platform for subcutaneous drug and fluid. See, for example, "Expert Opinion on Drug Delivery, 2007;4:427-440," in "Intradermal Administration," by Johns Hopkins University Press, 2007;4:427-440. The increased permeability of subcutaneous tissue due to the degradation of hyaluronan correlates with the effectiveness of hyaluronidases in their ability to increase the dispersion and absorption of compositions containing multiple particles administered simultaneously, sequentially, or intermittently.

[0214] In some embodiments, the composition further comprises at least one hyaluronan degrading agent, for example, a pharmaceutically effective amount of at least one hyaluronan degrading agent. In certain embodiments, the hyaluronan degrading agent is a hyaluronidase enzyme, also referred to herein as hyaluronidase. In some embodiments, the hyaluronidase is a soluble, neutral-active hyaluronidase. In certain embodiments, the hyaluronidase is a mammalian hyaluronidase. In certain embodiments, the mammalian hyaluronidase is a human hyaluronidase. In a preferred embodiment, the human hyaluronidase is a recombinant human hyaluronidase, such as rHuPH20. Certain recombinant human hyaluronidases suitable for use in the compositions and methods disclosed herein are commercially available, such as rHuPH20 from Halozyme Therapeutics (San Diego, CA).

[0215] According to certain embodiments of the present disclosure, the hyaluronan degrading agent can be administered simultaneously with the composition, sequentially, or intermittently. As used herein, the term "simultaneous" or "substantially simultaneous" administration refers to simultaneous administration of both components administered simultaneously, such as the composition and the hyaluronan degrading agent. In some embodiments, the hyaluronan degrading agent and the composition are administered in a single composition, e.g., co-formulated. In other embodiments, the hyaluronan degrading agent and the composition are administered as separate formulations. However, one component may be administered within minutes or hours, for example, in the same medical appointment or doctor's visit. Such administration is referred to as "sequential" administration. In certain embodiments, sequential administration refers to the sequential administration of the hyaluronan degrading agent and a composition as described herein. In preferred embodiments, the hyaluronan degrading agent is administered first, followed by one or more doses of the composition. In certain other embodiments, the components may be administered intermittently, for example, daily, weekly, or monthly, as a simultaneous formulation or separate formulations. In preferred embodiments, the hyaluronan degrading agent is administered first. For example, an initial higher loading dose, followed by one or more lower doses of the hyaluronan degrading agent and / or composition may be administered. However, other dosage regimens may be useful. The progress of these treatments can be easily monitored by conventional techniques and assays known to those skilled in the art.

[0216] The term "injectability" or "syringability" refers to the relative ease with which a liquid composition or formulation can be administered to a subject using an injection device. In particular, injectability is affected in part by the viscosity of the composition or formulation, the injection or transfer flow rate, and needle characteristics, such as length and gauge. In some embodiments, injectability is determined by measuring the viscosity of the composition or formulation at various shear rates. In other embodiments, injectability is determined by measuring the breakaway and / or glide forces required to actuate a standard injection device consisting of a barrel, plunger, and needle. In certain other embodiments, the syringe barrel has an internal diameter of at least 6 mm. As described herein, the injectability of a composition comprising a plurality of particles comprising at least one therapeutic biological agent is superior to that of an aqueous composition or formulation having approximately the same concentration of aqueous monomeric therapeutic biological agent. The term "injectability" or "syringability" may also refer to the ability to inject a pharmaceutical composition or formulation through a 16- to 33-gauge needle, optionally a thin-walled or ultra-thin-walled (UTW) syringe. In certain embodiments, the syringe is equipped with a needle that is at least 13 mm in length. Injectability is determined by factors such as the pressure or force required for injection, uniformity of flow, quality of aspiration, and lack of clogging. The injectability of a composition can be assessed by comparing the injection force of a reduced-viscosity composition with that of a standard composition without an added viscosity-lowering agent. Reduced-viscosity compositions as described herein can improve injectability, with the injection force reduced by at least 10%, preferably at least 30%, more preferably at least 50%, and most preferably at least 75% when compared to a standard composition with the same concentration of aqueous monomeric therapeutic biological agent under otherwise identical conditions. Alternatively, the injectability of a composition can be improved by reducing the injection force of a standard composition with the same concentration of aqueous monomeric therapeutic biological agent, for example, about 20% when the syringe is depressed with the same force. The flow rate can be assessed by comparing the time required to inject about 0.0 mL, preferably about 1.5 mL, more preferably about 1.0 mL, and most preferably about 0.5 mL of an aqueous monomeric therapeutic biological agent composition. The term "flow rate" refers to the volume of liquid that can pass through a given cross-sectional area per unit time. Generally, the formula for flow rate is Q = A x v, where Q is the flow rate, A is the cross-sectional area at a point in the flow path, and v is the average velocity of the liquid at that point. In certain embodiments, the flow rate is constant. In preferred embodiments, the flow rate is at least about 0.1 mL / sec.

[0217] The term "injection breakaway force" refers to the force required to overcome the friction between the syringe barrel and plunger of a standard injection device before ejection of the syringe contents can occur at a steady rate, e.g., the maximum force required to break the static friction of the plunger. The force is applied at the outward-facing end of the syringe plunger shaft and directed along the axis of the syringe barrel. The syringe contents are ejected through a syringe needle of a predetermined gauge and length. In certain embodiments, the injection breakaway force is measured by a load cell positioned at the outward-facing end of the syringe plunger during actuation.

[0218] The terms "injection glide force," "glide force," or "syringe force" refer to the force required to maintain steady ejection of the contents of a standard injection device, e.g., the force required to maintain plunger movement once static friction is overcome. The force is applied at the outward-facing end of the syringe plunger shaft and is directed along the axis of the syringe barrel. The syringe contents are expelled through a syringe needle of a given gauge and length. The term "Newtonian region" or "N" refers to a range of shear rates that are linearly or nearly linearly proportional to the local strain rate at every point.

[0219] In some embodiments, the composition is dispensed using a syringe force of about 2 N to about 80 N. In other embodiments, the composition is dispensed using a syringe force of about 2 N to about 40 N. In still other embodiments, the composition is dispensed using a syringe force of about 2 N to about 30 N. In certain other embodiments, the composition is dispensed using a syringe force of about 2 N to about 25 N. In certain embodiments, the composition may comprise a solubility limiting agent (SLU) of from about 2N to about 20N, e.g., from about 2N to about 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5, 15, 15.5, 16, 16.5, 17, 17.5, 18, 18.5, 19, 19.5, 20N; from about 2N to about 19N, e.g., from about 2N to about 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 8, 9, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5, 15, 15.5, 16, 16.5, 17, 17.5, 18, 18.5, 19, 19.5, 20N; 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5, 15, 15.5, 16, 16.5, 17, 17.5, 18, 18.5, 19N; about 2N to about 18N, for example, about 2N to about 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5, 15, 15.5, 16, 16. 5, 17, 17.5, 18N; about 2N to about 17N, for example, about 2N to about 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5, 15, 15.5, 16, 16.5, 17N; about 2N to about 16N, for example, about 2N to about 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11 1, 11.5, 12, 12.5, 13, 13.5, 14, 14.5, 15, 15.5, 16N; about 2N to about 15.5N, for example, about 2N to about 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5, 15, 15.5N; about 2N to about 15N, for example, about 2N to about 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8,8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5, 15N; about 2N to about 14.5N, for example, about 2N to about 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5N; about 2N to about 14N, for example, about 2N to about 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12. 5, 13, 13.5, 14N; about 2N to about 13.5N, for example, about 2N to about 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5N; about 2N to about 13N, for example, about 2N to about 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13N; about 2N to about 12.5N, for example, about 2N to about 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6 , 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5N; about 2N to about 12N, for example about 2N to about 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12N; about 2N to about 11.5N, for example about 2N to about 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5N; about 2N to about 11N, for example about 2N to about 2.5, 3, 3.5, 4, 4.5 , 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11N; about 2N to about 10.5N, for example, about 2N to about 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5N; about 2N to about 10N, for example, about 2N to about 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10N; about 2N to about 9.5N, for example, about 2N to about 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5N; about 2N to about 9N, for example, about 2N to about 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9N; about 2N to about 8.5N, for example, about 2N to about 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5N; about 2N to about 8N, for example, about 2N to about 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8N; about 2N to about 7.5N, for example, about 2N to about 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5N; about 2N to about 7N, for example, about 2N to about 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7N; about 2N to about 6.5N, for example, about 2N to about 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5N; about 2N to about 6N, for example, about 2N to about 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6N; about 2N to about 5.5N, for example, about 2N to about 2.5, 3, 3.5, 4, 4.5, 5, 5.5N; about 2N to about 5N, For example, the syringe force is about 2N to about 2.5, 3, 3.5, 4, 4.5, or 5 N; about 2N to about 4.5 N, for example, about 2N to about 2.5, 3, 3.5, 4, or 4.5 N; about 2N to about 4 N, for example, about 2N to about 2.5, 3, 3.5, or 4 N; or about 2N to about 3.5 N, for example, about 2N to about 2.5, 3, or 3.5 N. In a preferred embodiment, the syringe force is about 2N to about 15 N.

[0220] In other embodiments, the composition is dispensed using a syringe force of about 3 N to about 80 N. In some embodiments, the composition is dispensed using a syringe force of about 3 N to about 40 N. In still other embodiments, the composition is dispensed using a syringe force of about 3 N to about 30 N. In certain other embodiments, the composition is dispensed using a syringe force of about 3 N to about 25 N. In certain embodiments, the composition may comprise a solubility limiting agent (SLL) of about 3N to about 20N, e.g., about 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5, 15, 15.5, 16, 16.5, 17, 17.5, 18, 18.5, 19, 19.5 to about 20N; a solubility limiting agent (SLL) of about 3.5N to about 19N, e.g., about ... 4, 14.5, 15, 15.5, 16, 16.5, 17, 17.5, 18, 18.5 to about 19N; about 4N to about 15N, for example, about 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5 to about 15N; about 4N to about 14.5N, for example, about 4N to about 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5N; about 4N to about 14N, for example, about 4N to about 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14N; about 4N to about 13.5N, for example, about 4N to about 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8 , 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5N; about 4N to about 13N, for example about 4N to about 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13N; about 4N to about 12.5N, for example about 4N to about 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 1 The syringe force is about 4N to about 12N, e.g., about 4N to about 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12N; or about 4N to about 11.5N, e.g., about 4N to about 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12N. In a preferred embodiment, the syringe force is about 2N to about 25N. In another preferred embodiment, the syringe force is about 2N to about 20N. In a particularly preferred embodiment, the syringe force is about 2N to about 15N. In some embodiments, as the concentration of the therapeutic biological agent in the composition increases, the syringe force increases at a lower rate than the viscosity of the composition.

[0221] As shown in Figure 1, syringe force for a particle composition was studied using a 27-gauge ultra-thin-wall (UTW) needle with a 4.7 mm internal diameter syringe barrel. The results in Figure 1 indicate that the particle composition was dispensed using syringe forces up to approximately 11.5 N when the average sliding force was plotted against composition concentrations up to 600 mg / mL. The data was fitted using the Krieger-Dogherty model for concentrated suspensions of solid, spherical particles. 20 N (Newtons) was plotted as a reference to the typical sliding force limit. See Example 6. In other embodiments of the methods and compositions described herein, syringe force increases at a lower rate than the viscosity of the composition as the concentration of the therapeutic biological agent in the composition increases. As shown in Figure 7, the viscosity and syringe force of the particle composition (Elektroject) decrease as the protein concentration of the particle composition increases, compared to the viscosity and syringe force of the aqueous formulation, which remain proportional.

[0222] The therapeutic composition of the present embodiments is advantageously administered in the form of an injectable composition comprising a plurality of particles suspended in a low-viscosity pharmaceutically acceptable liquid carrier, each particle comprising at least one therapeutic biological agent or salt thereof. In some embodiments, the composition is administered by parenteral, subcutaneous, oral, epidermal, intradermal, intramuscular, intraarterial, intraperitoneal, or intravenous injection. In other embodiments, the composition is administered by subcutaneous injection. In certain embodiments, the composition is administered by syringe injection. In certain preferred embodiments, the composition is dispensed from a pre-filled syringe.

[0223] The therapeutic biological agents described herein can be formulated for parenteral administration, for example, by intravenous, intramuscular, subcutaneous, intravaginal, or even intraperitoneal injection. Typically, such compositions can be prepared as suspensions, and particle preparations can also be emulsified. In some embodiments, the compositions are administered by pre-filled devices, such as needleless injection devices, cartridges, or syringe injections. In other embodiments, the compositions are administered by subcutaneous syringe injections. In certain embodiments, the compositions is administered in one or more doses. In preferred embodiments, the composition is administered in a single dose. In certain other embodiments, the composition is administered in multiple doses.

[0224] Pharmaceutical compositions suitable for injectable use include sterile non-aqueous or aqueous liquid carriers; compositions containing sesame oil, peanut oil, corn oil, or aqueous propylene glycol; and sterile particles for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases, the composition must be sterile and fluid to the extent that easy syringability exists. It should also be stable under current good manufacturing practice (cGMP) and storage conditions and preserved against the contaminating action of microorganisms, such as bacteria and / or fungi.

[0225] Administering the methods described herein can include administering the composition using a 16-33 gauge needle. The 16-33 gauge needle can have a length of about 19 mm (¾ inch) or less; or preferably about 13 mm (½ inch) or less. In certain embodiments, administering the methods described herein can include administering the composition using a 27-33 gauge needle. In yet other embodiments, the 27-33 gauge needle has a length of about 13 mm (½ inch) or less. In other embodiments, the composition is dispensed from a 25-gauge to 33-gauge needle, e.g., a 26-, 27-, 28-, 29-, 30-, 31-, 32-gauge to 33-gauge needle; a 26-gauge to 33-gauge needle, e.g., a 27-gauge to 33-gauge needle, e.g., a 27-gauge to 33-gauge needle, e.g., a 28-gauge to 33-gauge needle, e.g., a 29-gauge to 33-gauge needle, e.g., a 30-gauge to 33-gauge needle, e.g., a 31-gauge to 33-gauge needle. In certain embodiments, the composition is dispensed from a 27-gauge to 30-gauge needle, e.g., a 28-, 29-gauge to 30-gauge needle. In a preferred embodiment, the composition is dispensed from a 27-gauge needle. In certain preferred embodiments, the composition is dispensed from a 30 gauge needle.

[0226] Administering any one of the methods described herein can include administering a dose of the composition to the subject in a single subcutaneous administration. In any one of the methods described herein, the method can further include co-administering to the subject at least one additional therapeutic agent, such as hyaluronidase, a cytotoxic or cytostatic agent, a nonsteroidal anti-inflammatory drug (NSAID), a pharmaceutically acceptable excipient, diluent, vehicle, carrier, or a combination thereof.

[0227] Compositions such as those described herein can be administered in vivo to human or animal subjects using a variety of known routes and techniques. For example, the compositions can be provided as injectable solutions, suspensions, or emulsions and administered via parenteral, subcutaneous, oral, epidermal, intradermal, intramuscular, intraarterial, intraperitoneal, or intravenous injection using a conventional needle and syringe. Alternatively, needle-free injection devices are suitable for administration. In some embodiments, the compositions are administered via syringe injection or needle-free injection. In other embodiments, the compositions are administered via needle-free injection. In certain other embodiments, the needle-free injection is a liquid jet injection. Needle-free injection encompasses a wide range of drug delivery systems that use Lorentz, shock wave, gas pressure, or electrophoretic forces to propel drugs through the skin, eliminating the use of hypodermic needles. The term "jet injection," as used herein, refers to a needle-free injection method in which a composition is forced through an orifice, thereby generating a high-pressure, ultra-fine liquid stream that can penetrate mammalian skin. In certain embodiments, jet injection is used for parenteral, subcutaneous, oral, epidermal, intradermal, intramuscular, intraarterial, intraperitoneal, or intravenous injection of a composition according to the present disclosure, hi a preferred embodiment, jet injection is used for subcutaneous injection.

[0228] In some embodiments, the composition is maintained for less than about 5 hours, e.g., less than 4 hours, after administration to the subject. Dissolves in less than 3 hours, less than 2 hours, or less than 1 hour. In other embodiments, the composition dissolves in less than about 60 minutes after administration to a subject, e.g., less than 59 minutes, less than 58 minutes, less than 57 minutes, less than 56 minutes, less than 55 minutes, less than 54 minutes, less than 53 minutes, less than 52 minutes, less than 51 minutes, less than 50 minutes, less than 49 minutes, less than 10 minutes, less than 9 minutes, less than 8 minutes, less than 7 minutes, less than 6 minutes, less than 5 minutes, less than 4 minutes, less than 3 minutes, less than 2 minutes, or less than 1 minute. In still other embodiments, the composition dissolves in less than about 20 minutes after administration to a subject. In certain embodiments, the composition dissolves in less than about 10 minutes after administration to a subject. In certain preferred embodiments, the composition dissolves in less than about 60 seconds, e.g., less than 59 seconds, less than 58 seconds, less than 57 seconds, less than 56 seconds, less than 55 seconds, less than 54 seconds, less than 53 seconds, less than 52 seconds, less than 51 seconds, less than 50 seconds, less than 49 seconds, ...less than 10 seconds, less than 9 seconds, less than 8 seconds, less than 7 seconds, less than 6 seconds, less than 5 seconds, less than 4 seconds, less than 3 seconds, less than 2 seconds, less than 1 second after administration to a subject.

[0229] In preferred embodiments according to the present disclosure described herein, when the composition is administered, complete dissolution occurs within a few seconds, which can reduce the immunological risk caused by particles remaining in the subcutaneous space. For example, compared to an equivalent dose of aqueous monomeric mAb, administration of particles containing mAb has been shown to produce similar pharmacokinetic (PK) profiles (AUC, Cmax, and Tmax) and efficacy (tumor growth reduction), as described herein. In certain preferred embodiments, the composition dissolves immediately after administration to a subject.

[0230] The method described herein demonstrates the subcutaneous delivery of highly concentrated Ab (300-400 mg / mL protein) compositions without loss of bioactivity. This was achieved for a variety of Abs by using ethyl oleate (EO) as the liquid carrier. Ethyl oleate is a fatty acid ester with a low viscosity of approximately 6 mPa·s at 25°C, allowing for rapid storage or administration. It does not chemically interact with the particles during dissolution in vivo. In some embodiments, ethyl oleate prevents aggregation and allows for easy resuspension of the particles prior to injection.

[0231] Dense (1.32 g / cm) with controllable size distribution (polydispersity index <0.2) 3 ) The production of round particles can be achieved using various Abs described herein. Focused ion beam scanning electron microscopy (FIB-SEM) can be used to determine whether particles contain void spaces, which is crucial for achieving high protein loading in the composition, and X-ray photoelectron spectroscopy (XPS) can be used to control the particle radial distribution. In certain embodiments, particle sizes can be approximately 5 to approximately 10 μm, which is preferable for achieving a low-viscosity composition but small enough to prevent syringe clogging with a 27-gauge needle. Adding particles to an ethyl oleate (EO) liquid carrier at approximately 400 mg / mL (protein concentration 380 mg / mL, stabilizing excipient 20 mg / mL) can form a therapeutic composition with a viscosity of approximately 20 mPa·s (correlated with a syringe force of approximately 4 N), which is useful for storage in prefilled syringes.

[0232] As described herein, characterization of the structural stability of particles in the compositions was achieved using size exclusion chromatography (SEC), differential scanning fluorescence (DSF), circular dichroism (CD), cation exchange chromatography (CIEX), and subvisible particle (SvP) analysis. Additionally, preservation of bioactivity was analyzed using flow cytometry and antibody-dependent cellular cytotoxicity (ADCC) assays.

[0233] Also described herein, SEC data confirmed that no aggregate formation was observed upon processing compared to the reformulated particle composition, which contained 1.8% aggregates, compared to the labeled formulation (aqueous mAb as the FDA-approved formulation), which contained 1.9% aggregates. After 30 days of storage at 40°C, DSF showed a thermal shift of less than 1°C / hr across the sample, and CD failed to detect any difference in secondary structure (beta-sheet percentage). CIEX was used to measure the solubility of the labeled formulation as required by regulation (ICH Q6B). Charged variants of such proteins were analyzed to ensure that chemical modification did not occur during the methods described herein and upon storage. In certain embodiments, the compositions may be less prone to chemical modification upon storage than FDA-labeled formulations. This is because the proteins are more stable in the solid state as particles.

[0234] After 30 days of storage at 40°C, flow cytometry assays demonstrated preservation of bioactivity. No discernible differences were apparent between the FDA-labeled formulations and their compositions as described herein. ADCC assays were also performed to assess the binding and effector function of each Ab for its target by exposure to Ab-incubated target cell lines. As shown in Figures 2A and 2B, bioactivity significantly decreased after storage for the FDA-labeled formulations, while no decrease in activity was observed for the particles in the compositions.

[0235] Prior to the initiation of in vivo studies, the particles used for the compositions were obtained aseptically. Bacterial endotoxin levels were approximately three orders of magnitude lower than accepted injection standards (0.05 EU / mg to 0.25 EU / mg). Furthermore, microbial growth assays showed no observable growth.

[0236] The compositions described herein are comparable to aqueous FDA-labeled formulations in terms of rat pharmacokinetics (PK), SC clearance (mice), and efficacy (mouse xenografts). PK profiles are shown in Figure 3 for the aqueous mAb formulation (SC injection) and the mAb composition (SC) in Sprague-Dawley rats (n=5 for each condition) and were comparable and within the well-accepted bioequivalence limits of 80-125%. Because undissolved particles can potentially cause an immunogenic response, the in vivo dissolution behavior of the particles has been shown to ensure that the compositions can clear from the injection site at a similar rate compared to standard aqueous formulations. See Example 7.

[0237] By conjugating a fluorescent dye to rituximab, it was possible to visualize particle dissolution in vivo in the upper dorsal region of SKH1 (hairless mice), as shown in Figure 4. See Example 9. Similar clearance by fluorescent signal was observed for both the aqueous formulation and the particle composition. After 14 days, the animals were sacrificed, and five major organs (liver, spleen, kidney, lung, and heart) were collected, homogenized, and fluorescence quantified. Negligible fluorescence was observed, indicating that rituximab was completely metabolized and that particles were not transported to the animal's major organs. Efficacy studies relied on measuring DOHH-2 xenograft tumor size in Fox Chase SCID mice treated with the aqueous formulation and particle composition. As expected, Figure 5 shows that tumor growth was prevented with the particle composition and was comparable to the aqueous monomeric mAb formulation. See Example 8. Furthermore, there was no substantial toxicity or other pathology at any of the injection sites or in any major organs of the injected mice.

[0238] Further in vivo studies were extended to hematology, serum chemistry, and PK in minipigs. Hematology and serum chemistry measurements were shown to fall within normal ranges with no significant changes in serum chemistry (day 3) or hematology (day 7) markers for either group. Pilot PK profiles (n=2 for each condition) show similar bioavailability in Yucatan minipigs, whose subcutaneous space closely resembles that of humans, as shown in Figure 6. See Example 10.

[0239] kit In various embodiments, the method comprises administering to the subject a therapeutic biological agent comprising ... Kits are contemplated. For example, the kit may include one or more additional containers, each holding one or more of various materials (e.g., reagents and / or equipment, optionally in concentrated form) desirable from a commercial and user standpoint for use of the compositions described herein. Non-limiting examples of such materials include, but are not limited to, buffers, diluents, filters, needles, syringes, cartridges; labels for carriers, packages, containers, vials, and / or tubes listing the contents and / or instructions for use, and package inserts with instructions for use. In some embodiments, the present embodiments contemplate kits for administering the therapeutics of the embodiments. The kit may include one or more sealed, pre-filled syringes or cartridges containing any of the pharmaceutical compositions of the present disclosure. The kit may include, for example, at least one antibody and reagents for preparing, compounding, and / or administering components of the embodiments or for performing one or more steps of the disclosed treatment methods. In other embodiments, the kit may also include suitable containers that do not react with the components of the kit, such as Eppendorf tubes, syringes, bottles, or tubes. The containers may be made from a material that can be sterilized, such as plastic or glass. In a preferred embodiment, the composition is dispensed from a pre-filled syringe.

[0240] The kit may further include an instruction sheet outlining the procedural steps of the methods described herein, following substantially the same procedures as described herein or known to those of skill in the art. The instructional information may be in a computer-readable medium containing computer-readable instructions that, when executed using a computer, result in a representation of an actual or virtual procedure for delivering a pharmaceutically effective amount of a therapeutic biological agent.

[0241] A label is optionally on or associated with a container. For example, a label is on a container when letters, numbers, or other symbols forming the label are affixed, stamped, or etched into the container itself; a label is associated with a container when present in a receptacle or carrier that also holds the container, e.g., as a package insert. Furthermore, a label can be used to indicate that the contents are to be used for a specific therapeutic application. Furthermore, a label can provide instructions for using the contents, for example, in the methods described herein. In certain embodiments, the pharmaceutical composition is present in a pack or dispenser device containing one or more unit dosage forms containing a therapeutic biological agent provided herein. For example, the pack contains metal or plastic foil, e.g., a blister pack. Alternatively, the pack or dispenser device can be accompanied by instructions for administration. Alternatively, the pack or dispenser can have notice associated with the container in a form prescribed by a government agency regulating the manufacture, use, or sale of pharmaceuticals, the notice reflecting approval by the agency of the drug form for human or veterinary administration. Such notice, for example, is the labeling approved by the U.S. Food and Drug Administration for prescription drugs or an approved product insert. In some embodiments, compositions containing a therapeutic biological agent provided herein formulated in a compatible pharmaceutical liquid carrier are prepared, placed in an appropriate container, and labeled for treatment of an indicated condition.

[0242] In some cases, the liquid compositions provided herein are formulated in a pre-filled injection device, such as a syringe or needleless injection. In some embodiments, the liquid compositions are formulated in a volume of about 0.1 mL, 0.2 mL, 0.3 mL, 0.4 mL, 0.5 mL, 0.6 mL, 0.7 mL, 0.8 mL, 0.9 mL, 1.0 mL, 1.1 mL, 1.2 mL, 1.3 mL, 1.4 mL, 1.5 mL, 1.6 mL, 1.7 mL, 1.8 mL, 1.9 mL, 2.0 mL, 2.1 mL, 2.2 mL, 2.3 mL, 2.4 mL, or about 2.5 mL. In other embodiments, the volume of the administered composition is about 2.5 mL. In certain embodiments, the volume of the administered composition is about 2.0 mL, preferably about 1.5 mL, more preferably about 1.0 mL, and most preferably about 0.5 mL. In a preferred embodiment, the volume of the composition dispensed from the pre-filled syringe is from about 1.5 mL to about 2.0 mL.

[0243] In other cases, the composition further comprises a pharmaceutically effective amount of at least one hyaluronan degrading agent, such as a hyaluronidase. In some embodiments, the administered composition, e.g., a composition formulated with at least one hyaluronan degrading agent, has a volume of less than about 2.0 L, e.g., less than about 1.8 L, 1.5 L, 1.2 L, 1.0 L, 0.8 L, 0.5 L, 0.3 L, 0.1 L, or less than about 800 mL, 700 mL, 600 mL, 500 mL, 400 mL, 300 mL, 200 mL, 100 mL, 90 mL, 80 mL, 70 mL, 60 mL, 50 mL, 40 mL, 30 mL, 20 mL, 10 mL, 9.0 mL, 8.0 mL, 7.0 mL, 6.0 mL, 5.0 mL, 4.0 mL, 3.0 mL, or less than about 2.5 mL. In other embodiments, the administered composition is less than about 20.0 mL. In certain embodiments, the composition administered is less than about 10.0 mL. In certain other embodiments, the composition administered is less than about 5.0 mL.

[0244] In some embodiments, such kits comprise a pre-filled injection device of the present disclosure, such as a syringe or needle-free injector, in a blister pack. The blister pack may itself be sterile inside. In other embodiments, a pre-filled injection device according to the present disclosure, such as a syringe or needle-free injector, may be placed in such a blister pack before undergoing sterilization, such as terminal sterilization.

[0245] The present disclosure having generally been described will be more readily understood by reference to the following examples, which are included solely for purposes of illustration, and not limitation, of certain aspects and embodiments of the present disclosure.

[0246] Example Abbreviation Å Angstrom aa amino acid BSA Bovine serum albumin ℃ Celsius temperature cm centimeters d day DCM dichloromethane DIPEA Diisopropylethylamine DMA N,N-dimethylaniline DMF Dimethylformamide DMSO dimethyl sulfoxide DTE Dithioerythritol DTT Dithiothreitol EDT 1,2-ethanedithiol EDTA Ethylenediaminetetraacetic acid eq. equivalent weight Et Ethyl g grams h time HPLC High Performance Liquid Chromatography Hz Hertz IV (intravenous) kJ kilojoule LC-MS Liquid Chromatography Mass Spectrometry m Meta mAb monoclonal antibody MALDI-MS matrix-assisted laser desorption / ionization mass spectrometry Me methyl MHz Megahertz min μg microgram μL microliter μm micrometer μM micromolar mg milligram mL milliliter mm millimeters mM millimolar mol mole nm nanometer NMP N-methylpyrrolidone p Para PBS Phosphate Buffered Saline PEG polyethylene glycol PEGA Polyethylene glycol polyacrylamide ppm parts per million ps picoseconds RP-HPLC Reversed-phase high-performance liquid chromatography rpm Revolutions per minute s seconds SC subcutaneous sec second SEM Scanning Electron Microscopy t Third grade tert third grade UHMW Ultra High Molecular Weight Polyethylene ug micrometer UTW Ultra Thin Wall UV ultraviolet light V-Bolt vol% Volume percent wt% weight percent

[0247] immunotherapy Those skilled in the art will understand that immunotherapy can be used in conjunction with the methods of the embodiments described herein. In the context of treating a disease or condition, such as cancer, an inflammatory disease or condition, an immune disorder, a renal disease, a skin disease or condition, or a human immunodeficiency virus (HIV) infection, in a subject in need thereof, immunotherapeutics generally rely on the use of immune effector cells and therapeutic biological agents or immune effectors to target and treat the disease or condition, such as cancer, an inflammatory disease or condition, an immune disorder, a renal disease, or a skin disease or condition, or a human immunodeficiency virus (HIV) infection. Rituximab (RITUXAN®) is one such example. The immune effector can be, for example, an antibody specific for some marker on the surface of the target cell. The antibody can serve as a therapeutic effector or recruit other cells to actually affect the cell. Various effector cells include, but are not limited to, cytotoxic T cells and NK cells.

[0248] In some embodiments, the methods described herein feature methods of treating a patient by administering to the patient a composition for subcutaneous administration, e.g., a composition comprising a plurality of particles comprising at least one therapeutic biological agent in a composition as described herein. In certain embodiments, the patient has a disease or condition. In preferred embodiments, the composition is administered as a single dose by subcutaneous syringe injection. In other embodiments, the patient has cancer, an inflammatory disease or condition, an immune disease, a renal disease, or a skin disease or condition, or human immunodeficiency virus (HIV) infection.

[0249] In certain embodiments of immunotherapy, tumor cells should bear several markers that are amenable to targeting, i.e., that are not present on the majority of other cells. In the context of this embodiment, there are many tumor markers that may be suitable for targeting. Common tumor markers include, but are not limited to, CD20, carcinoembryonic antigen, tyrosinase (p97), gp68, TAG-72, HMFG, sialyl Lewis antigen, MucA, MucB, PLAP, laminin receptor, erb B, or p155. An alternative aspect of immunotherapy is to combine anti-cancer effects with immunostimulatory effects. There are also immunostimulatory molecules, including, but not limited to, cytokines such as IL-2, IL-4, IL-12, GM-CSF, gamma-IFN, chemokines such as MIP-1, MCP-1, and IL-8, and growth factors such as FLT3 ligand. Examples of immunotherapies currently under investigation or in use include immune adjuvants such as Mycobacterium bovis, Plasmodium falciparum, dinitrochlorobenzene, and aromatic compounds (U.S. Pat. Nos. 5,801,005 and 5,739,169; Hui and Hashimoto, 1998; Christodoulides et al., 1998); cytokine therapy such as interferon α, β, and γ, IL-1, GM-CSF, and TNF (Bukowski et al., 1998; Davidson et al., 1998; Hellstrand et al., 1998); gene therapy such as TNF, IL-1, IL-2, and p53 (Qin et al., 1998; Austin-Ward and and monoclonal antibodies, such as anti-CD20, anti-ganglioside GM2, and anti-p185 (Hollander, 2012; Hanibuchi et al., 1998; U.S. Patent No. 5,824,311). It is contemplated that one or more anti-cancer therapies may be used in conjunction with the antibody therapies described herein.

[0250] material The Roche rituximab biosimilar was purchased from a distributor that provided the antibody in an aqueous composition consistent with the FDA label, specified as 10 mg / mL rituximab, 9 mg / mL sodium chloride, 7.35 mg / mL sodium citrate dihydrate, and 0.7 mg / mL polysorbate 80. The application-specific "feed solution" composition used to process the particles was generated by modifying the FDA-labeled formulation by desalting, followed by concentration and addition of the desired excipients. Human IgG was obtained from Equitech-Bio Inc., Kerrville, TX.

[0251] Cell Culture: DOHH2 human B-cell lymphoma cells (DSMZ) were maintained in Roswell Park Memorial Institute (RPMI) medium containing 10% fetal bovine serum (FBS) and 1% penicillin G / streptomycin.

[0252] Animals: Female wild-type albino Sprague-Dawley rats (Charles R The animals (Liver Lab) were housed in the animal facility at Tufts University Comparative Medicine Services (Tufts CMS). All described animal use was in accordance with the National Research Council's Guide for the care and use of laboratory animals and was performed according to detailed written protocols approved by the Institutional Animal Care and Use Committee (IACUC).

[0253] CR female CB.17 SCID mice (Charles River Labs) were housed in the Charles River Labs animal facility. All animal procedures were officially reviewed and approved by the Charles River Labs Institutional Animal Care and Use Committee (IACUC).

[0254] Male (castrated) Yucatan pigs (S&S Farms, Constantine, MI) were housed in the animal facility at CBSET, Inc. All animal procedures were officially reviewed and approved by the Institutional Animal Care and Use Committee (IACUC) at CBSET, Inc.

[0255] Female wild-type Balb / c mice (Charles River Labs) were housed in the animal facility at Tufts University Comparative Medicine Services (Tufts CMS). All animal procedures were officially reviewed and approved by the Institutional Animal Care and Use Committee (IACUC) at Tufts CMS.

[0256] As is known in the art, therapeutically effective amounts for use in humans can also be determined from animal models. For example, a dose for humans can be formulated to achieve a concentration found to be effective in animals. The dosage in humans can be adjusted by monitoring the effectiveness of the therapeutic biological agent and adjusting the dosage upward or downward, as described above. Adjusting the dosage to achieve maximum efficacy in humans based on the above and other methods is well within the capabilities of one skilled in the art. Dosages can be determined based on the IC as determined in cell culture in animal models. 50 The drug can be formulated to achieve a concentration range including (the concentration of drug that achieves half-maximal inhibition of symptoms). Such information can be used to more accurately determine useful doses in humans. Levels in blood or tissue samples can be measured, for example, by high performance liquid chromatography (HPLC). Generally, the dose equivalent of a drug is about 1 ng / kg to about 100 mg / kg for a typical subject.

[0257] Tumor xenograft model: DOHH2 human B-cell lymphoma cells were harvested at 70% confluency and administered subcutaneously (SC) to the flanks of female CB.17 SCID mice. 1 x 10 DOHH-2 tumor cells in 0% Matrigel were injected at a cell injection volume of 0.1 mL / mouse. Tumors were allowed to grow to between 80 and 120 mm before treatment began. 3 When the average size of went.

[0258] method FlowCam: Particle size measurements were measured using FlowCam; a dynamic image analysis instrument. Samples were diluted to approximately 1 mg / mL in isopropanol and passed through a thin channel. Images of the particles were recorded and analyzed by size and shape (number weighted).

[0259] ImageJ Measurement: Particle diameter was measured using ImageJ analysis on SEM images. Analysis was performed on 600x images. The ImageJ particle analysis tool was run on the images to identify objects with a circularity of >0.8 and a size >0.5um, along with the outline of each object. These contours were visually inspected for fit. Misidentified particles were manually rejected, and missing particles were manually included and measured using the ImageJ diameter tool.

[0260] Accelerated Storage Protocol: All samples were transferred to Wheaton EZ Extraction round-bottom glass vials (2 mL or 4 mL volume depending on the sample) for aging. The glass vials were sealed with parafilm, placed in an oven at 40°C, and visually inspected daily over the aging period to ensure integrity.

[0261] Viscosity measurements: Unless otherwise noted, suspension viscosity was measured at 25 °C using an AR-G2 rheometer (TA Instruments) and a 25 mm plate. Measurements were performed at 1000 s (experimental limit due to edge effects), which is below the shear rate experienced in a 27-gauge needle but within the Newtonian region for the suspension. Each measurement was repeated three times (with approximately 60 s between repeats) to assess the short-term physical stability of the suspension. Calibration standards were recorded before each measurement to verify instrument settings. Viscosity can also be calculated from the syringe force by applying the Hagen-Poiseuille equation using the suspension flow rate through the needle.

[0262] Injectability measurements: Unless otherwise stated, syringe forces were measured during 0.1 mL / s ejection of 1 mL of suspension (400 mg / mL particles) using a custom-built force sensor device and a 1 mL Norm-ject model syringe with a 27-gauge ultra-thin-walled needle (TSK).

[0263] Karl Fischer: Tests for moisture content were performed using Karl Fischer analysis: approximately 100 mg of particles were heated to 105° C. in an oven and the released water was determined coulometrically.

[0264] Skeletal density: Skeletal density was measured by gas pycnometry. The gas can be nitrogen or an equivalent gas, and the particle mass can be 0.0413 g.

[0265] Particle dissolution: Phosphate-buffered saline (PBS) was added to the dried particle sample to produce a final concentration of 10 mg / mL (particle mass / mL of solution). The sample was placed on a VWR angle rocker with a speed setting of "35" and an angle setting of "15." At 1, 10, 20, 30, 40, 50, 60, 90, and 120 minutes, 10 μL aliquots were removed from the sample vial and the absorbance at 280 nm was measured and recorded. For all samples, Ab, e.g., mAb, concentration was plotted against time.

[0266] Salt content: Salt content was recorded by measuring sodium content using inductively coupled plasma optical emission spectroscopy (ICP-OES). A calibration curve was prepared using a sodium standard (ICP TraceCERT, 1000 mg / L). Quality control was completed using a standard solution diluted at 100 ppm sodium. A sample of particles (approximately 15 mg) dissolved in 2% by volume nitric acid (10 mL) was then analyzed, resulting in an intensity below the instrument's detection limit of approximately 0.5 ppm for sodium. This indicated a sodium content of less than 0.034% by weight and a total salt content of less than 0.1% by weight (assuming equal removal of sodium citrate and sodium chloride).

[0267] Size exclusion chromatography (SEC) measurements: 20 μL injections of sample (1 mg / mL) were run on an Agilent AdvanceBio SEC (300 mm × 2.7 μm, 300 Å column) in SEC buffer (25 mM phosphate, 250 mM NaCl, pH 6.8) at a flow rate of 1 mL / min for 15 min. Peak analysis was performed by automatic integration using the following parameters: gradient sensitivity = 0.5, peak width = 0, height reject = 0, area reject = 0, shoulder off, area percent reject = 0, standard tangent. Vent skim mode, advanced baseline correction, front peak skim height ratio 0, tail peak skim height ratio 0, peak valley ratio 0 and skim valley ratio 0.

[0268] Differential scanning fluorimetry (DSF) measurements: Protein melting temperatures before and after formulation and at various time points during storage at 40°C were assessed using a QuantStudio6Flex instrument. Five microliters (5 μL) of sample (1 mg / mL) prepared after dialysis was added in quadruplicate to a 96-well thermal cycler plate. To each well, 12.5 μL of ultrapure deionized water and 2.5 μL of SYPRO® Orange dye (8×) were added. After a 5-minute incubation, the samples were run from 25°C to 99°C at a ramp rate of 0.05°C / s. Melting temperatures were calculated using Protein Thermal Shift software (Thermo Fisher, version 1.3) using a Boltzmann fit.

[0269] Circular dichroism (CD) measurements: The degree of preservation of the secondary structure (alpha helix and beta sheet) of the protein before and after formulation and at various times during storage at 40°C was measured using Jasco Evaluation was performed using a J-815 instrument. Four hundred microliters (400 μL) of sample (0.5 mg / mL) prepared after dialysis was added to a quartz cuvette (1 mm path length). The sample was scanned over the 190-260 nm range. Diluent buffer was used as a blank subtraction for each sample. The following instrument settings were used: Metering mode: CD, HT Measurement range: 260~190nm Data pitch: 0.5nm Sensitivity: Standard DIT:4 seconds Bandwidth: 1.00nm Start Mode: Immediate Scanning speed: 100 nm / min Shutter control: Automatic Baseline correction: None CD detector: PMT PMT voltage: Automatic

[0270] Cation Exchange Chromatography (CIEX) Measurements: Charge variant analysis was performed on each sample under accelerated storage conditions on days 0, 7, and 30 using an Agilent BioMAb NP5, 4.6 x 250 mm, PEEK ion exchange column. Samples were prepared at a concentration of 1 mg / mL after overnight dialysis in water. Buffer A was prepared with 30 mM phosphate, pH 6.3, and 0 mM NaCl. Buffer B was prepared with Buffer A, 30 mM phosphate, pH 6.3, and 175 mM NaCl. Samples were run in a gradient that started with 100% Buffer A and ramped to 100% Buffer B over a 20-minute period, then returned to 100% Buffer A and 0% Buffer B over the next minute. The system was re-equilibrated in 100% Buffer A for 10 minutes before the next sample was injected. Integration was performed in manual skim peak mode, and Agilent data was reflected in the following protocol: https: / / www.agilent.com / cs / library / applications / 5991-5557EN.pdf.

[0271] Flow cytometry: One million Raji cells (100 μL per well) were plated per well in a 96-well "V-bottom" plate, and 10 μL of mAb, label, particles, or suspension was added to the well for a starting concentration of 200 μL. The dilution factor for all antibodies was 3x. The plates were incubated at 4°C for 30 minutes. The plates were centrifuged at 2000 rpm for 5 minutes and washed three times with PBS. 100 μL of PE-conjugated goat anti-human IgG was added as a secondary antibody at a 1:200 dilution. The plates were centrifuged at 2000 rpm for 5 minutes and washed three times with PBS. The plates were then centrifuged at 2000 rpm for 5 minutes and washed three times with PBS. The plates were then incubated at 4°C for 30 minutes. The plates were centrifuged at 2000 rpm for 5 minutes and washed three times with PBS. 100 μL of PE-conjugated goat anti-human IgG was added as a secondary antibody at a 1:200 dilution. The plates were then centrifuged at 2000 rpm for 5 minutes and washed three times with PBS. The plates were then incubated at 4°C for 30 minutes. The plates were centrifuged at 2000 rpm for 5 minutes and washed three times with PBS. The plates were then incubated at 4°C for 30 minutes. The plates were centrifuged at 2000 rpm for 5 minutes and washed three times with PBS. The plates were then incubated at 4°C for 30 minutes. The plates were then ... Cells were resuspended in 200 μL of cold PBS for analysis on a Technologies Attune NXT flow cytometer.

[0272] Scanning Electron Microscopy: Electron micrographs were collected on selected samples on a Hitachi TM3030Plus or TM1000 tabletop microscope. Samples were immobilized on conductive tape and examined in a low-vacuum anti-static environment, eliminating the need for sample preparation.

[0273] Image Analysis: Selected microscopy images were selected for further analysis based on (i) minimal particle overlap, (ii) sufficient contrast between particles and background, and (iii) a resolution that achieved particle occupancy of at least 10 pixels. This allowed particles to be easily identified and reduced resolution-based errors. A binarization threshold was applied to separate particles from the background, and a watershed segmentation algorithm was applied to ensure that individual particles were measured separately. The ImageJ tool "Analyze Particles" was then applied to the binary images with the following parameters: circularity between 0.5 and 1.0; size between 5 and infinity square microns; edge exclusion; and hole filling. The outlines of identified particles were superimposed on the original image. Misidentified particles, such as single particles whose outlines did not match a particle or clusters identified as particles, were then discarded. Missing particles were measured by manually tracing the particle outline and using ImageJ's measurement tools.

[0274] Density Analysis: The skeletal density of particles from selected samples was determined by testing approximately 0.1 g of powder with an AccuPyc II 1340 gas displacement pycnometer system.

[0275] Moisture content analysis: Residual moisture in particles from selected samples was determined by placing approximately 0.1 g of powder in an oven with a Karl Fischer titrator and heating the sample.

[0276] ELISA assay: An ELISA assay was used on selected samples to detect human antibodies in a denaturing-sensitive format. Human IgG was first plated in PBS for 1 hour, followed by three 4-minute washes with wash buffer (PBS + 0.05% Tween 20), followed by blocking with 2% BSA (Sigma) in wash buffer for 45 minutes, followed by incubation with dilute (20 μg / mL) Protein A-HRP (Abcam) for 45 minutes, followed by three 3-minute washes with wash buffer, followed by incubation with TMB (Abcam) for 10 minutes, and finally quenching the reaction with stop solution (Abcam). Colorimetric readings were performed on a Thermo Multiskan Spectrum.

[0277] Subvisible Particle (SvP) Analysis: Subvisible particles (SvP) were analyzed using a Fluid Imaging Technologies FlowCam PV-100 system. Samples for analysis were reconstituted to the desired concentration with filtered water (Milli-Q) in sterile centrifuge tubes. Three sets of samples were then examined. These included (i) a sample of the diluent used for reconstitution, (ii) a sample of an aliquot of the feed solution, i.e., the first liquid, used for the particle formation process, and (iii) the reconstituted material.

[0278] Accelerated Storage: Unless otherwise stated, storage is performed on selected samples by incubating them. Accelerated conditions were performed by maintaining the samples in a rotor or oven at elevated temperature (40°C) for a defined period of time. Samples were kept in 2 mL or 4 mL Wheaton glass vials and sealed with paraffin film.

[0279] Helium ion microscopy (HIM): Ion micrographs were collected on selected samples using an HIM instrument. The source energy, working distance, and aperture size were typically 29 keV, 9 mm, and 10 microns, respectively. For selected samples, a focused gallium ion beam was used to section particles for analysis of their internal structure. Tilt samples were ablated with a source current, dwell time, and section interval of 300 pA, 0.5-1 μs, and 2-5 nm, respectively.

[0280] X-ray photoelectron spectroscopy (XPS): A small amount of powder was deposited on hydrocarbon tape attached to a piece of silicon wafer and gently pressed to form a compact, uniform bed. Excess loose powder was removed by gently tapping the edge of the wafer piece. Specimens were prepared immediately prior to analysis. XPS measurements were performed on a Kratos Axis Ultra spectrometer using monochromated AlKα X-rays (1486.6 eV). For ea...

Claims

[Claim 1] The invention described in the specification.

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