Pharmaceutical compositions of dupilumab

Abstract

Disclosed herein, in part, are pharmaceutical compositions comprising dupilumab and arginine-glutamate and methods of using the pharmaceutical compositions to treat medical disorders e.g., atopic dermatitis, eosinophilic esophagitis, prurigo nodularis, asthma, or chronic rhinosinusitis in a patient in need thereof.

Description

Attorney Docket No. TPH-001WOPHARMACEUTICAL COMPOSITIONS OF DUPILUMABCROSS-REFERENCE TO RELATED APPLICATION

[0001] This application claims the benefit of and priority to U.S. Provisional Patent Application No. 63 / 713,470, filed October 29, 2024, which is incorporated by reference in its entirety herein.BACKGROUND

[0002] Dupilumab is a recombinant human IgG4 monoclonal antibody (mAb) which inhibits interleukin 4 (IL-4) and interleukin 13 (IL-13) signaling. Dupilumab is currently approved for the treatment of atopic dermatitis in adult and pediatric patients, eosinophilic esophagitis in adult and pediatric patients, and prurigo nodularis in adult patients. Dupilumab is also currently approved as an add-on maintenance treatment of asthma in adult and pediatric patients and chronic rhinosinusitis with nasal polyposis in adult patients.

[0003] There is a need for pharmaceutical compositions of dupilumab which are sufficiently stable and suitable for administration to patients.SUMMARY

[0004] The invention provides pharmaceutical compositions comprising dupilumab and methods of using the pharmaceutical compositions to treat medical disorders e.g., atopic dermatitis, eosinophilic esophagitis, prurigo nodularis, asthma, or chronic rhinosinusitis in a patient in need thereof.

[0005] One aspect of the invention provides a pharmaceutical composition comprising dupilumab and arginine-glutamate. In some aspects, the pharmaceutical composition comprises dupilumab, arginine-glutamate, and proline. In some aspects, the pharmaceutical composition comprises dupilumab, arginine-glutamate, and sucrose. Another aspect of the invention provides a pharmaceutical composition comprising dupilumab, polysorbate, proline, and arginine-glutamate. Another aspect of the invention provides a pharmaceutical composition comprising dupilumab, polysorbate, sucrose, and arginine-glutamate. In some embodiments, the pharmaceutical composition has a pH of about 5.9. In some embodiments, the pharmaceutical composition is a liquid. In some embodiments, dupilumab has a concentration of about 100 mg / mL to about 200 mg / mL. In some embodiments, dupilumab has a concentration of about 175 mg / mL. In some embodiments, dupilumab has a concentration of about 150 mg / mL In some embodiments, arginine- glutamate has a concentration of about 50 mM. In some embodiments, proline has a concentrationAttorney Docket No. TPH-001WO of about 150 mM. In some embodiments, sucrose has a concentration of about 150 mM. In some embodiments, polysorbate has a concentration of about 0.2 % (w / v). Another aspect of the invention provides a pharmaceutical composition comprising about 175 mg / mL dupilumab, about 2 mg / mL polysorbate, about 150 mM proline, and about 50 mM arginine-glutamate. Another aspect of the invention provides a pharmaceutical composition comprising about 175 mg / mL dupilumab, about 2 mg / mL polysorbate, about 150 mM sucrose, and about 50 mM arginineglutamate. In some embodiments, the pharmaceutical composition is substantially free of histidine. In some embodiments, the pharmaceutical composition is substantially free of acetate. In some embodiments, the pharmaceutical composition has a viscosity of about 15 to 17 cP. In some embodiments, the pharmaceutical composition has a Tmi of about 66 °C to 68 °C and a Tm2 of about 72 °C to 73 °C. In some embodiments, the pharmaceutical composition has an osmolality of about 300 to 350 mOsm / kg. In some embodiments, the pharmaceutical composition has a polydispersity index (PDI) of about 0.17 to 0.19. In some embodiments, the pharmaceutical composition has a hydrodynamic radius (Rh) of about 9.5 nm.BRIEF DESCRIPTION OF THE FIGURES

[0006] FIG. 1 depicts protein concentration (left) and aggregation index (right) in the control samples (2-8 °C after 1 month) and the stressed samples (40 °C after 1 month). Horizontal lines are the averages, and shaded areas are the 95% confidence intervals.

[0007] FIG. 2 depicts the hydrodynamic radius (Rh) and polydispersity index (PDI) in the control samples (2-8 °C after 1 month) and the stressed samples (40 °C after 1 month). Results are organized based on buffer system. Horizontal lines are the averages, and shaded areas are the 95% confidence intervals within the buffer system.

[0008] FIG. 3 depicts the aggregation onset temperature (Taggr) by DLS in the control samples (2-8 °C after 1 month) and the stressed samples (40 °C after 1 month). Results are organized based on the buffer system. Horizontal lines are the averages, and shaded areas are the 95% confidence intervals within the buffer system.

[0009] FIG. 4 depicts the %Main, %HMW and %LMW by SEC in the control samples (2- 8 °C after 1 month) and the stressed samples (40 °C after 1 month). Results are organized based on buffer system. Horizontal lines are the averages, and shaded areas are the 95% confidence intervals within the buffer system.2IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WO

[0010] FIG. 5 depicts the percentage of main, acidic, and basic isoforms by icIEF in the control samples (2-8 °C after 1 month) and the stressed samples (40 °C after 1 month). Results are organized based on the buffer system. Horizontal lines are the averages, and shaded areas are the 95% confidence intervals within the buffer system.

[0011] FIG. 6 depicts pairwise correlations of %main, %acidic, and %basic charge variants for the stressed samples (40 °C pL after 1 month).

[0012] FIG. 7 depicts protein concentration of the control samples (TZ, 1 month 2-8 °C), after 1 month at 40 °C , and after three freeze-thaw cycles (FT). The solid line shows target concentration (175 mg / mL) and the dashed line shows the target + 10% concentration (192.5 mg / mL).

[0013] FIG. 8 depicts aggregation index (Al) of the control samples (TZ, 1 month 2-8 °C), after 1 month at 40 °C , and after three freeze-thaw cycles (FT).

[0014] FIG. 9 depicts hydrodynamic radius (top) and polydispersity index (bottom) of the control samples (TZ, 1 month 2-8 °C), after 1 month at 40 °C (40C), and after three freeze-thaw cycles (FT).

[0015] FIG. 10 depicts diffusion interaction parameter ko in the formulations.

[0016] FIG. 11 depicts %Monomer (top), %HMW (middle), and %LMW (bottom) in the control samples (TZ, 1 month 2-8 °C), after 1 month at 40 °C (40C), and after three freeze-thaw cycles (FT). The line shows the values measured in the reference formulation. The other line shows the values measured in the reference formulation after 1 month at 40 °C (40C).

[0017] FIG. 12 depicts %Main (top), %Acidic (middle), and %Basic (bottom) isoforms in the control samples (TZ, 1 month 2-8 °C), after 1 month at 40 °C (40C), and after three freeze-thaw cycles (FT). The solid line shows the values measured in the reference formulation. The other line shows the values measured in the reference formulation after 1 month at 40 °C (40C).

[0018] FIG. 13 depicts thermal unfolding onset (Tonset), and midpoints (Tml, Tm2, and Tm4) for the formulations for initial samples.

[0019] FIG. 14 depicts solution viscosity at 20 °C for the formulations.

[0020] FIG. 15 depicts calculated viscosity at 175 mg / mL at 20°C for the formulations.The line shows viscosity of 15.4 cP (protein in the reference formulation).

[0021] FIG. 16 depicts protein concentration for samples at time 0, 1 month at 25 °C and 40 °C, 3 months at 2-8 °C, 25 °C, and 40 °C, 6 months at 2-8 °C and 25 °C.

[0022] FIG. 17 depicts Al for the stability samples at time 0, stored at 2-8 °C for 3 and 6 months, at 25 °C for 1, 3, and 6 months, and at 40 °C for 1 and 3 months.3IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WO

[0023] FIG. 18 depicts polydispersity index (PDI) for the formulations and 200 mg / 1.14 mL for the reference product (175 mg / mL).

[0024] FIG. 19 depicts SEC-%HMW for the stability samples.

[0025] FIG. 20 depicts SEC Chromatograms for the stability samples (panels A, B, C, andD).

[0026] FIG. 21 depicts SEC-%Monomer for the stability samples.

[0027] FIG. 22 depicts SEC-%HMW for the stressed samples.

[0028] FIG. 23 depicts example icIEF Electropherograms for the stability samples (initial and 6M 25 °C ).

[0029] FIG. 24 depicts the stability trend of the %Main charge variants for the stability samples at 2-8 °C, 25 °C and 40 °C .

[0030] FIG. 25 depicts the stability trend of the %Acidic charge variants for the stability samples at 2-8 °C, 25 °C and 40 °C .

[0031] FIG. 26 depicts icIEF electropherograms for the 3-month stability samples at 40 °C.

[0032] FIG. 27 depicts the stability trend of the %Basic charge variants for the stability samples at 2-8 °C, 25 °C and 40 °C .

[0033] FIG. 28 depicts example NRCE-SDS electropherograms for the stability samples, (initial and 6M 25 °C).

[0034] FIG. 29 depicts the stability trend of the purity (%IgG by NRCE-SDS) for the stability samples at 2-8 °C, 25 °C and 40 °C .

[0035] FIG. 30 depicts example RCE-SDS electropherograms for the stability samples (initial and 6M 25 °C)

[0036] FIG. 31 depicts the DSC for the formulations: (A) example deconvolution of the thermogram showing unfolding temperature (Tonset), thermal transition for CH2 (Tml), CH3 (Tm2), and Fab (Tm4) domains; (B) thermograms for the formulations and the reference product.

[0037] FIG. 32 depicts the viscosity profile for the formulations at 20 °C .DETAILED DESCRIPTION

[0038] The invention provides pharmaceutical compositions comprising dupilumab and methods of using said pharmaceutical compositions to treat medical disorders e.g., atopic dermatitis, eosinophilic esophagitis, prurigo nodularis, asthma, or chronic rhinosinusitis in a patient4IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WO in need thereof. In particular, the invention provides pharmaceutical compositions comprising dupilumab and arginine-glutamate.Definitions

[0039] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Abbreviations used herein have the conventional meanings as known within the chemical and biological arts. The chemical structures and formulae set forth herein are constructed according to the standard rules of chemical valency known in the chemical arts.

[0040] The terms “a” and “an” as used herein mean “one or more” and include the plural unless the context is inappropriate.

[0041] In the application, where an element or component is said to be included in and / or selected from a list of recited elements or components, the element or component can be any one of the recited elements or components, or the element or component can be selected from a group consisting of two or more of the recited elements or components.

[0042] Further, elements and / or features of a composition or a method described herein can be combined in a variety of ways without departing from the spirit and scope of the present invention. For example, where reference is made to a particular compound, that compound can be used in various embodiments of compositions of the present invention and / or in methods of the present invention, unless otherwise understood from the context. Within this application, embodiments have been described and depicted in a way that enables a clear and concise application, but embodiments may be combined or separated without parting from the present teachings and invention(s).

[0043] The expression “at least one of’ includes individually each of the recited objects after the expression as well as the various combinations of two or more of the recited objects unless otherwise understood from the context. The expression “and / or” in connection with recited objects should be understood to have the same meaning unless otherwise understood from the context.

[0044] The use of the term “include,” “includes,” “including,” “have,” “has,” “having,” “contain,” “contains,” “containing,” and grammatical equivalents thereof, are open-ended and nonlimiting, unless otherwise specifically stated or understood from the context.

[0045] Where the use of the term “about” is before a quantitative value, the present invention also includes the specific quantitative value itself unless specifically stated otherwise. As used herein, the term “about” refers to a ±10% variation from the nominal value unless otherwise indicated or inferred from the context.5IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WO

[0046] At various places in the present specification, values are disclosed in groups or in ranges. It is specifically intended that the description include each and every individual member of such groups and ranges. For example, an integer in the range of 0 to 40 is specifically intended to individually disclose 0, 1, 2, 3, 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, and 40, and an integer in the range of 1 to 20 is specifically intended to individually disclose 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20.

[0047] The use of any and all examples, or exemplary language herein (for example, “such as” or “including,”) is intended merely to illustrate better the present invention and does not pose a limitation on the scope of the invention unless claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the present invention.

[0048] As used herein, the term “substantially free” means that either no substance is present or only minimal, trace amounts of the substance are present which do not have any substantial impact on the properties of the composition. If reference is made to no amount of a substance, it should be understood as “no detectable amount.”

[0049] As used herein, a “compound” (including a specifically named compound) refers to the compound itself and its pharmaceutically acceptable salts unless otherwise understood from the context of the description or expressly limited to one particular compound or form.

[0050] As used herein, the term “pharmaceutical composition” refers to a medicinal or pharmaceutical formulation that contains an active ingredient as well as one or more excipients and diluents to enable the active ingredient suitable for the method of administration. The pharmaceutical composition of the present disclosure includes pharmaceutically acceptable components that are compatible with dupilumab.

[0051] As used herein, the terms “pharmaceutical composition” and “formulation” are used interchangeably.

[0052] As used herein, “dupilumab” is synonymous with the active pharmaceutical ingredient in Dupixent®. Dupilumab has a chemical formula of C6512H10066N1730O2052S46 and a molecular weight of approximately 147 kilodaltons. Dupilumab has a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 1 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:5 as disclosed in U.S. Patent Number 11,059,896. The term “dupilumab” is also intended to encompass so-called biosimilar or biobetter variants of the dupilumab protein used in commercially available Dupixent®. For example, a variant of commercial Dupixent® may be acceptable to the FDA when it has essentially the same6IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WO pharmacological effects as commercially available Dupixent®, even though it may exhibit certain physical properties that may be similar if not identical to Dupixent®.

[0053] For the purposes of the present application, the term “dupilumab” also encompasses dupilumab with minor modifications in the amino acid structure (including deletions, additions, and / or substitutions of amino acids) or in the glycosylation properties, which do not significantly affect the function of the polypeptide.

[0054] The term “antibody,” as used herein, is generally intended to refer to immunoglobulin molecules comprising four polypeptide chains, two heavy (H) chains and two light (L) chains inter-connected by disulfide bonds, as well as multimers thereof (e.g., IgM). Each heavy chain comprises a heavy chain variable region (abbreviated herein as HCVR or VH) and a heavy chain constant region. The heavy chain constant region comprises three domains, CHI, CH2 and CH3. Each light chain comprises a light chain variable region (abbreviated herein as LCVR or VL) and a light chain constant region. The light chain constant region comprises one domain (CL1). The Vnand VL regions can be further subdivided into regions of hypervariability, termed complementarity determining regions (CDRs), interspersed with regions that are more conserved, termed framework regions (FR). Each Vnand Vris composed of three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.

[0055] Unless specifically indicated otherwise, the term “antibody,” as used herein, shall be understood to encompass complete antibody molecules as well as antigen-binding fragments thereof. The term “antigen-binding portion” or “antigen-binding fragment” of an antibody (or simply “antibody portion” or “antibody fragment”), as used herein, refers to one or more fragments of an antibody that retain the ability to specifically bind to a particular epitope.

[0056] An “isolated antibody,” as used herein, is intended to refer to an antibody that is substantially free of other antibodies having different antigenic specificities (e.g., an isolated antibody that specifically binds hIL-4Ra is substantially free of antibodies that specifically bind antigens other than hIL-4Ra).

[0057] As used herein, the terms “subject” and “patient” refer to organisms to be treated by the methods and / or compositions described herein. Such organisms are preferably mammals (e.g., murines, simians, equines, bovines, porcines, canines, felines, and the like), and more preferably humans.

[0058] As used herein, a “buffer” refers to an aqueous solution that is resistant to changes in pH. A buffer may include a “buffering agent” such as a weak acid and its salt, or a weak base and its salt, which assist in maintaining the stability of the pH. Examples of buffers used in7IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WO pharmaceutical formulations include bicarbonate buffers, carbonate buffers, citrate buffers, histidine buffers, phosphate buffers, tartrate buffers, tris(hydroxymethyl)aminomethane (or 2- amino-2-hydroxymethyl-propane-l,3-diol [(HOCH2)3CNH2]) buffers, and combinations thereof. Some of these buffers are suitable for pharmaceutical formulations administered subcutaneously.

[0059] As used herein, “tonicity” refers to the ionic strength or concentration of ions in a solution such as a pharmaceutical formulation. Tonicity often is measured in molarity (“M”). As used herein, an “isotonic solution,” an “isotonic formulation,” an “isotonic pharmaceutical formulation,” and a pharmaceutical formulation that is “isotonic” refers to a solution or formulation that has the same or similar concentration of ions as found in bodily fluids.

[0060] As used herein, “physiological pH” refers to a pH of about 7.4.

[0061] As used herein, “osmoticity” and “osmolality” refer to the osmotic pressure of a solution such as a pharmaceutical formulation. Osmoticity often is measured in osmolarity (“Osm / L” or “OsM”) or osmolality (“Osm / kg”), which can be used interchangeably herein. When measuring freezing point depression, the observed value is the osmolality of the solution. In contrast to tonicity, osmoticity accounts for un-ionized solutes in a solution such that when present, the osmolarity or osmolality of the solution will be higher than its tonicity. The osmolarity of a pharmaceutical composition described herein can be measured, for example, using a vapor pressure method.

[0062] As used herein, an “isosmotic solution,” an “isosmotic formulation,” an “isosmotic pharmaceutical formulation,” and a pharmaceutical formulation that is “isosmotic” refers to a solution or a formulation that has the same or similar concentration of solutes as found in bodily fluids. In some embodiments, a pharmaceutical composition that is “isosmotic” can have an osmolarity in the range of about 275 mOsM to about 350 mOsM.

[0063] As used herein, “osmolarity adjustor” and “osmotic agent” refer to a pharmaceutically acceptable compound that may be added to a pharmaceutical composition described herein in order to modulate the osmolarity of the pharmaceutical composition.

[0064] As used herein, “pharmaceutically acceptable” refers to a substance that is acceptable for use in pharmaceutical applications from a toxicological perspective and does not adversely interact with the active ingredient. Accordingly, pharmaceutically acceptable carriers are those that are compatible with the other ingredients in the formulation and are biologically acceptable. In some embodiments, supplementary active ingredients can also be incorporated into the pharmaceutical compositions.

[0065] As used herein, “pharmaceutically acceptable excipient” refers to any substance other than the active agent that is included in the compositions of the present invention. Non-8IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WO limiting examples of pharmaceutically acceptable excipients include water, NaCl, normal saline solutions, a phosphate buffered saline solution, emulsions (e.g., such as an oil / water or water / oil emulsions), lactated Ringer’s, normal sucrose, normal glucose, binders, fillers, disintegrants, lubricants, coatings, sweeteners, flavors, salt solutions (such as Ringer’s solution), alcohols, oils, gelatins, carbohydrates such as lactose, amylose or starch, fatty acid esters, hydroxymethylcellulose, polyvinyl pyrrolidone, and colors, and the like. Such preparations can be sterilized and, if desired, mixed with auxiliary agents such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, coloring, and / or aromatic substances and the like that do not deleteriously react with the compounds of the invention. For examples of excipients and carriers, see Martin, Remington’s Pharmaceutical Sciences, 15th Ed., Mack Publ. Co., Easton, PA (1975).

[0066] As used herein, the term “pharmaceutically acceptable carrier” refers to any of the standard pharmaceutical carriers, such as a phosphate buffered saline solution, water, emulsions (e.g., such as an oil / water or water / oil emulsions), and various types of wetting agents. The compositions also can include stabilizers and preservatives. For examples of carriers, stabilizers and adjuvants, see Martin, Remington's Pharmaceutical Sciences, 15th Ed., Mack Publ. Co., Easton, PA

[1975] ,

[0067] As used herein, the term “pharmaceutically acceptable salt” refers to any pharmaceutically acceptable salt (e.g., acid or base) of a compound of the present invention which, upon administration to a subject, is capable of providing a compound of this invention or an active metabolite or residue thereof. As is known to those of skill in the art, “salts” of the compounds of the present invention may be derived from inorganic or organic acids and bases. Examples of acids include, but are not limited to, hydrochloric, hydrobromic, sulfuric, nitric, perchloric, fumaric, maleic, phosphoric, glycolic, lactic, salicylic, succinic, toluene-p-sulfonic, tartaric, acetic, citric, methane sulfonic, ethanesulfonic, formic, benzoic, malonic, naphthalene-2-sulfonic, benzene sulfonic acid, and the like. Other acids, such as oxalic, while not in themselves pharmaceutically acceptable, may be employed in the preparation of salts useful as intermediates in obtaining the compounds of the invention and their pharmaceutically acceptable acid addition salts.

[0068] Examples of bases include, but are not limited to, alkali metal (e.g., sodium) hydroxides, alkaline earth metal (e.g., magnesium) hydroxides, ammonia, and compounds of formula NWE, wherein W is Ci-4 alkyl, and the like.

[0069] Examples of salts include, but are not limited to: acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptanoate,9IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WO glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, oxalate, palmoate, pectinate, persulfate, phenylpropionate, picrate, pivalate, propionate, succinate, tartrate, thiocyanate, tosylate, undecanoate, and the like. Other examples of salts include anions of the compounds of the present invention compounded with a suitable cation such as Na+, NH4. and NW4+(wherein W is a Ci-4 alkyl group), and the like.

[0070] As used herein, the term “effective amount” refers to the amount of a composition (e.g., a pharmaceutical composition of the present invention) sufficient to effect beneficial or desired results. An effective amount can be administered in one or more administrations, applications or dosages and is not intended to be limited to a particular formulation or administration route.

[0071] As used herein, the terms “treat,” “treating,” and “treatment” include any effect, e.g., lessening, reducing, modulating, ameliorating or eliminating, that results in the improvement of the condition, disease, disorder, and the like, or ameliorating a symptom thereof.

[0072] The phrase "therapeutically-effective amount" as used herein means that amount of a composition (e.g., a pharmaceutical composition of the present invention) which is effective for producing some desired therapeutic effect in a subject.

[0073] As used herein, the term “stable” refers to the ability of the antibody to maintain its therapeutic efficacy (e.g., all or the majority of its intended biological activity and / or physiochemical integrity) over extended periods of time. The stability of the antibody, and the capability of the pharmaceutical composition to maintain stability of such antibody, may be assessed over extended periods of time (e.g., for at least 1, 3, 6, 12, 18, 24, 30, 36 months or more). In general, pharmaceutical compositions described herein have been formulated such that they are capable of stabilizing, or alternatively slowing or preventing the degradation, of one or more antibody formulated therewith. In the context of a formulation a stable formulation is one in which the antibody therein essentially retains its physical and / or chemical integrity and biological activity upon storage and during processes (such as freeze / thaw, mechanical mixing and lyophilization). In some embodiments, protein stability may be measured by formation of high molecular weight (HMW) aggregates, loss of activity, generation of peptide fragments and shift of charge profdes.

[0074] Throughout the description, where compositions and kits are described as having, including, or comprising specific components, or where processes and methods are described as having, including, or comprising specific steps, it is contemplated that, additionally, there are compositions and kits of the present invention that consist essentially of, or consist of, the recited10IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WO components, and that there are processes and methods according to the present invention that consist essentially of, or consist of, the recited processing steps.

[0075] As a general matter, compositions specifying a percentage are by weight unless otherwise specified. Further, if a variable is not accompanied by a definition, then the previous definition of the variable controls.Pharmaceutical Compositions of Dupilumab

[0076] As described herein, in one aspect, the invention provides pharmaceutical compositions comprising dupilumab.

[0077] In various embodiments, the pharmaceutical composition generally comprises dupilumab and one or more pharmaceutically acceptable excipients.

[0078] In some embodiments, the pharmaceutical composition comprises a buffer. In some embodiments, the buffer comprises one, two, three, or four amino acids. In some embodiments, the amino acid is a naturally occurring amino acid. In some embodiments, the amino acid is selected from the 22 amino acids found in humans (alanine, arginine, asparagine, aspartate, cysteine, glutamine, glutamate, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine, etc.).

[0079] In some embodiments, the pharmaceutical composition comprises glutamate. In some embodiments, the pharmaceutical composition comprises arginine. In some embodiments, the pharmaceutical composition comprises proline. In some aspects, the pharmaceutical composition comprises arginine -glutamate. In some aspects, the pharmaceutical composition comprises dupilumab, glutamate, arginine, and proline, and additional excipients.

[0080] In some embodiments, the pharmaceutical composition comprises dupilumab, glutamate, arginine, a sugar-based excipient, and other excipients. In some embodiments, the sugar-based excipient is a disaccharide. In other embodiments, the sugar-based excipient is a monosaccharide. In some embodiments, the sugar-based excipient is dextrin, dextrose, fructose, fucose, galactose, glucose, glycogen, starch, maltose, lactose, mannose, ribose, sucrose, trehalose, or xylose. In some embodiments, the sugar-based excipient is sucrose. In some embodiments, the pharmaceutical composition comprises dupilumab, glutamate, arginine, sucrose, and other excipients.

[0081] In some aspects, pharmaceutical compositions described herein comprise a nonionic surfactant. In some aspects, pharmaceutical compositions described herein comprise an emulsifier. In some aspects, pharmaceutical compositions described herein comprise a nonionic surfactant, and11IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WO the nonionic surfactant is polysorbate. In some embodiments, the polysorbate is polysorbate 80 (PS80), or polysorbate 20 (PS20). In some embodiments, the polysorbate is polysorbate 80 (PS80).

[0082] In some embodiments, the pharmaceutical composition comprises dupilumab, glutamate, arginine, proline, polysorbate, and other excipients. In some embodiments, the pharmaceutical composition comprises dupilumab, glutamate, arginine, sucrose, polysorbate, and other excipients.

[0083] In various embodiments, the concentration of the one or more pharmaceutically acceptable excipients in the pharmaceutical compositions described herein can be from about 25 mM to about 500 mM, from about 30 mM to about 500 mM, from about 40 mM to about 500 mM, from about 50 mM to about 500 mM, from about 75 mM to about 500 mM, from about 100 mM to about 500 mM, from about 150 mM to about 500 mM, from about 200 mM to about 500 mM, from about 250 mM to about 500 mM, from about 300 mM to about 500 mM, from about 350 mM to about 500 mM, from about 400 mM to about 500 mM, from about 450 mM to about 500 mM, from about 25 mM to about 450 mM, from about 25 mM to about 400 mM, from about 25 mM to about 350 mM, from about 25 mM to about 300 mM, from about 25 mM to about 250 mM, from about 25 mM to about 200 mM, from about 25 mM to about 150 mM, from about 25 mM to about 100 mM, from about 25 mM to about 75 mM, from about 25 mM to about 50 mM, from about 25 mM to about 40 mM, from about 25 mM to about 30 mM, from about 30 mM to about 450 mM, from about 30 mM to about 400 mM, from about 30 mM to about 350 mM, from about 30 mM to about 300 mM, from about 30 mM to about 250 mM, from about 30 mM to about 200 mM, from about 30 mM to about 150 mM, from about 30 mM to about 100 mM, from about 30 mM to about 75 mM, from about 30 mM to about 50 mM, from about 30 mM to about 40 mM, from about 40 mM to about 450 mM, from about 40 mM to about 400 mM, from about 40 mM to about 350 mM, from about 40 mM to about 300 mM, from about 40 mM to about 250 mM, from about 40 mM to about 200 mM, from about 40 mM to about 150 mM, from about 40 mM to about 100 mM, from about 40 mM to about 75 mM, from about 40 mM to about 50 mM, from about 50 mM to about 450 mM, from about 50 mM to about 400 mM, from about 50 mM to about 350 mM, from about 50 mM to about 300 mM, from about 50 mM to about 250 mM, from about 50 mM to about 200 mM, from about 50 mM to about 150 mM, from about 50 mM to about 100 mM, from about 50 mM to about 75 mM, from about 75 mM to about 450 mM, from about 75 mM to about 400 mM, from about 75 mM to about 350 mM, from about 75 mM to about 300 mM, from about 75 mM to about 250 mM, from about 75 mM to about 200 mM, from about 75 mM to about 150 mM, from about 75 mM to about 100 mM, from about 100 mM to about 450 mM, from about 100 mM to12IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WO about 400 mM, from about 100 mM to about 350 mM, from about 100 mM to about 300 mM, from about 100 mM to about 250 mM, from about 100 mM to about 200 mM, from about 100 mM to about 150 mM, from about 150 mM to about 450 mM, from about 150 mM to about 400 mM, from about 150 mM to about 350 mM, from about 150 mM to about 300 mM, from about 150 mM to about 250 mM, from about 150 mM to about 200 mM, from about 200 mM to about 450 mM, from about 200 mM to about 400 mM, from about 200 mM to about 350 mM, from about 200 mM to about 300 mM, from about 200 mM to about 250 mM, from about 250 mM to about 450 mM, from about 250 mM to about 400 mM, from about 250 mM to about 350 mM, from about 250 mM to about 300 mM, from about 300 mM to about 450 mM, from about 300 mM to about 400 mM, from about 300 mM to about 350 mM, from about 350 mM to about 450 mM, from about 350 mM to about 400 mM, or from about 400 mM to about 450 mM. In some embodiments, the concentration of the one or more pharmaceutically acceptable excipients in the pharmaceutical composition is from about 25 mM to about 250 mM. In some embodiments, the concentration of the one or more pharmaceutically acceptable excipients in the pharmaceutical composition is from about 50 mMto about 150 mM.

[0084] In various embodiments, the concentration of arginine-glutamate in the pharmaceutical compositions described herein can be from about 25 mM to about 250 mM, from about 50 mM to about 250 mM, from about 75 mM to about 250 mM, from about 100 mM to about 250 mM, from about 125 mM to about 250 mM, from about 150 mM to about 250 mM, from about 175 mM to about 250 mM, from about 200 mM to about 250 mM, from about 225 mM to about 250 mM, from about 25 mM to about 225 mM, from about 25 mM to about 200 mM, from about 25 mM to about 175 mM, from about 25 mM to about 150 mM, from about 25 mM to about 125 mM, from about 25 mM to about 100 mM, from about 25 mM to about 75 mM, from about 25 mM to about 50 mM, from about 50 mM to about 225 mM, from about 50 mM to about 200 mM, from about 50 mM to about 175 mM, from about 50 mM to about 150 mM, from about 50 mM to about 125 mM, from about 50 mM to about 100 mM, from about 50 mM to about 75 mM, from about 75 mM to about 225 mM, from about 75 mM to about 200 mM, from about 75 mM to about 175 mM, from about 75 mM to about 150 mM, from about 75 mM to about 125 mM, from about 75 mM to about 100 mM, from about 100 mM to about 225 mM, from about 100 mM to about 200 mM, from about 100 mM to about 175 mM, from about 100 mM to about 150 mM, from about 100 mM to about 125 mM, from about 125 mM to about 225 mM, from about 125 mM to about 200 mM, from about 125 mM to about 175 mM, from about 125 mM to about 150 mM, from about 150 mM to about 225 mM, from about 150 mM to about 200 mM, from about 150 mM to about 175 mM, from about 150 mM to about 250 mM, from about 175 mM to about 225 mM, from about 17513IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WO mM to about 200 mM, or from about 200 mM to about 225 mM. In some embodiments, the concentration of arginine-glutamate in the pharmaceutical composition is from about 25 mM to about 250 mM. In some embodiments, the concentration of arginine-glutamate in the pharmaceutical composition is from about 25 mM to about 100 mM.

[0085] In some embodiments, the concentration of arginine-glutamate in the pharmaceutical compositions described herein can be about 25 mM, about 50 mM, about 75 mM, about 100 mM, about 125 mM, about 150 mM, about 175 mM, about 200 mM, about 225 mM, or about 250 mM. In some embodiments the concentration of arginine-glutamate in the pharmaceutical compositions is about 50 mM.

[0086] In various embodiments, the concentration of proline in the pharmaceutical compositions described herein can be from about 25 mM to about 250 mM, from about 50 mM to about 250 mM, from about 75 mM to about 250 mM, from about 100 mM to about 250 mM, from about 125 mM to about 250 mM, from about 150 mM to about 250 mM, from about 175 mM to about 250 mM, from about 200 mM to about 250 mM, from about 225 mM to about 250 mM, from about 25 mM to about 225 mM, from about 25 mM to about 200 mM, from about 25 mM to about 175 mM, from about 25 mM to about 150 mM, from about 25 mM to about 125 mM, from about 25 mM to about 100 mM, from about 25 mM to about 75 mM, from about 25 mM to about 50 mM, from about 50 mM to about 225 mM, from about 50 mM to about 200 mM, from about 50 mM to about 175 mM, from about 50 mM to about 150 mM, from about 50 mM to about 125 mM, from about 50 mM to about 100 mM, from about 50 mM to about 75 mM, from about 75 mM to about 225 mM, from about 75 mM to about 200 mM, from about 75 mM to about 175 mM, from about 75 mM to about 150 mM, from about 75 mM to about 125 mM, from about 75 mM to about 100 mM, from about 100 mM to about 225 mM, from about 100 mM to about 200 mM, from about 100 mM to about 175 mM, from about 100 mM to about 150 mM, from about 100 mM to about 125 mM, from about 125 mM to about 225 mM, from about 125 mM to about 200 mM, from about 125 mM to about 175 mM, from about 125 mM to about 150 mM, from about 150 mM to about 225 mM, from about 150 mM to about 200 mM, from about 150 mM to about 175 mM, from about 150 mM to about 250 mM, from about 175 mM to about 225 mM, from about 175 mM to about 200 mM, or from about 200 mM to about 225 mM. In some embodiments, the concentration of proline in the pharmaceutical composition is from about 25 mM to about 250 mM. In some embodiments, the concentration of proline in the pharmaceutical composition is from about 25 mM to about 100 mM.

[0087] In some embodiments, the concentration of proline in the pharmaceutical compositions described herein can be about 25 mM, about 50 mM, about 75 mM, about 100 mM,14IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WO about 125 mM, about 150 mM, about 175 mM, about 200 mM, about 225 mM, or about 250 mM. In some embodiments the concentration of proline in the pharmaceutical compositions is about 150 mM.

[0088] In various embodiments, the concentration of sucrose in the pharmaceutical compositions described herein can be from about 25 mM to about 250 mM, from about 50 mM to about 250 mM, from about 75 mM to about 250 mM, from about 100 mM to about 250 mM, from about 125 mM to about 250 mM, from about 150 mM to about 250 mM, from about 175 mM to about 250 mM, from about 200 mM to about 250 mM, from about 225 mM to about 250 mM, from about 25 mM to about 225 mM, from about 25 mM to about 200 mM, from about 25 mM to about 175 mM, from about 25 mM to about 150 mM, from about 25 mM to about 125 mM, from about 25 mM to about 100 mM, from about 25 mM to about 75 mM, from about 25 mM to about 50 mM, from about 50 mM to about 225 mM, from about 50 mM to about 200 mM, from about 50 mM to about 175 mM, from about 50 mM to about 150 mM, from about 50 mM to about 125 mM, from about 50 mM to about 100 mM, from about 50 mM to about 75 mM, from about 75 mM to about 225 mM, from about 75 mM to about 200 mM, from about 75 mM to about 175 mM, from about 75 mM to about 150 mM, from about 75 mM to about 125 mM, from about 75 mM to about 100 mM, from about 100 mM to about 225 mM, from about 100 mM to about 200 mM, from about 100 mM to about 175 mM, from about 100 mM to about 150 mM, from about 100 mM to about 125 mM, from about 125 mM to about 225 mM, from about 125 mM to about 200 mM, from about 125 mM to about 175 mM, from about 125 mM to about 150 mM, from about 150 mM to about 225 mM, from about 150 mM to about 200 mM, from about 150 mM to about 175 mM, from about 150 mM to about 250 mM, from about 175 mM to about 225 mM, from about 175 mM to about 200 mM, or from about 200 mM to about 225 mM. In some embodiments, the concentration of sucrose in the pharmaceutical composition is from about 25 mM to about 250 mM. In some embodiments, the concentration of sucrose in the pharmaceutical composition is from about 25 mM to about 100 mM.

[0089] In some embodiments, the concentration of sucrose in the pharmaceutical compositions described herein can be about 25 mM, about 50 mM, about 75 mM, about 100 mM, about 125 mM, about 150 mM, about 175 mM, about 200 mM, about 225 mM, or about 250 mM. In some embodiments the concentration of sucrose in the pharmaceutical compositions is about 150 mM.

[0090] In various embodiments, the concentration of dupilumab in the pharmaceutical compositions described herein can be from about 50 mg / mL to about 250 mg / mL, from about 60 mg / mL to about 250 mg / mL, from about 70 mg / mL to about 250 mg / mL, from about 80 mg / mL to15IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WO about 250 mg / mL, from about 90 mg / mL to about 250 mg / mL, from about 100 mg / mL to about 250 mg / mL, from about 120 mg / mL to about 250 mg / mL, from about 140 mg / mL to about 250 mg / mL, from about 160 mg / mL to about 250 mg / mL, from about 180 mg / mL to about 250 mg / mL, from about 200 mg / mL to about 250 mg / mL, from about 50 mg / mL to about 200 mg / mL, from about 50 mg / mL to about 180 mg / mL, from about 50 mg / mL to about 160 mg / mL, from about 50 mg / mL to about 140 mg / mL, from about 50 mg / mL to about 120 mg / mL, from about 50 mg / mL to about 100 mg / mL, from about 50 mg / mL to about 90 mg / mL, from about 50 mg / mL to about 80 mg / mL, from about 50 mg / mL to about 70 mg / mL, from about 50 mg / mL to about 60 mg / mL, from about 60 mg / mL to about 200 mg / mL, from about 60 mg / mL to about 180 mg / mL, from about 60 mg / mL to about 160 mg / mL, from about 60 mg / mL to about 140 mg / mL, from about 60 mg / mL to about 120 mg / mL, from about 60 mg / mL to about 100 mg / mL, from about 60 mg / mL to about 90 mg / mL, from about 60 mg / mL to about 80 mg / mL, from about 60 mg / mL to about 70 mg / mL, from about 70 mg / mL to about 200 mg / mL, from about 70 mg / mL to about 180 mg / mL, from about 70 mg / mL to about 160 mg / mL, from about 70 mg / mL to about 140 mg / mL, from about 70 mg / mL to about 120 mg / mL, from about 70 mg / mL to about 100 mg / mL, from about 70 mg / mL to about 90 mg / mL, from about 70 mg / mL to about 80 mg / mL, from about 80 mg / mL to about 200 mg / mL, from about 80 mg / mL to about 180 mg / mL, from about 80 mg / mL to about 160 mg / mL, from about 80 mg / mL to about 140 mg / mL, from about 80 mg / mL to about 120 mg / mL, from about 80 mg / mL to about 100 mg / mL, from about 80 mg / mL to about 90 mg / mL, from about 90 mg / mL to about 200 mg / mL, from about 90 mg / mL to about 180 mg / mL, from about 90 mg / mL to about 160 mg / mL, from about 90 mg / mL to about 140 mg / mL, from about 90 mg / mL to about 120 mg / mL, from about 90 mg / mL to about 100 mg / mL, from about 100 mg / mL to about 200 mg / mL, from about 100 mg / mL to about 180 mg / mL, from about 100 mg / mL to about 160 mg / mL, from about 100 mg / mL to about 140 mg / mL, from about 100 mg / mL to about 120 mg / mL, from about 120 mg / mL to about 200 mg / mL, from about 120 mg / mL to about 180 mg / mL, from about 120 mg / mL to about 160 mg / mL, from about 120 mg / mL to about 140 mg / mL, from about 140 mg / mL to about 200 mg / mL, from about 140 mg / mL to about 180 mg / mL, from about 140 mg / mL to about 160 mg / mL, from about 160 mg / mL to about 200 mg / mL, from about 160 mg / mL to about 180 mg / mL, or from about 180 mg / mL to about 200 mg / mL. In some embodiments, the concentration of dupilumab in the pharmaceutical composition is from about 50 mg / mL to about 250 mg / mL, from about 60 mg / mL to about 250 mg / mL, from about 70 mg / mL to about 250 mg / mL, or from about 80 mg / mL to about 250 mg / mL. In some embodiments the concentration of dupilumab in the pharmaceutical composition is from about 80 mg / mL to about 250 mg / mL.16IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WO

[0091] In various embodiments, the concentration of dupilumab in the pharmaceutical compositions described herein can be greater than about 40 mg / mL, greater than about 50 mg / mL, greater than about 60 mg / mL, greater than about 70 mg / mL, greater than about 80 mg / mL, greater than about 90 mg / mL, greater than about 100 mg / mL, greater than about 120 mg / mL, greater than about 140 mg / mL, greater than about 160 mg / mL, greater than about 175 mg / mL, greater than about 180 mg / mL, greater than about 200 mg / mL, greater than about 220 mg / mL, or greater than about 250 mg / mL. In some embodiments, the concentration of dupilumab in the pharmaceutical composition is greater than about 40 mg / mL. In some embodiments, the concentration of dupilumab in the pharmaceutical composition is greater than about 50 mg / mL. In some embodiments, the concentration of dupilumab in the pharmaceutical composition is greater than about 60 mg / mL. In some embodiments the concentration of dupilumab in the pharmaceutical composition is greater than about 70 mg / mL. In some embodiments the concentration of dupilumab in the pharmaceutical composition is greater than about 80 mg / mL.

[0092] In various embodiments the concentration of dupilumab in the pharmaceutical compositions described herein can be about 40 mg / mL, about 50 mg / mL, about 60 mg / mL, about 70 mg / mL, about 80 mg / mL, about 90 mg / mL, about 100 mg / mL, about 120 mg / mL, about 140 mg / mL, about 160 mg / mL, about 175 mg / mL, about 180 mg / mL, about 200 mg / mL, about 220 mg / mL, or about 250 mg / mL. In some embodiments, the concentration of dupilumab in the pharmaceutical composition is about 175 mg / mL.In various embodiments, the concentration of polysorbate in the pharmaceutical compositions described herein can be about 0.01 % (w / v) to about 50 % (w / v), e.g., about 0.01 % (w / v) to about 10 % (w / v), about 0.01 % (w / v) to about 5 % (w / v), e.g., about 0.01 % (w / v) to about 4 % (w / v), about 0.01 % (w / v) to about 3 % (w / v), about 0.01 % (w / v) to about 2 % (w / v), about 0.01 % (w / v) to about 1 % (w / v), about 0.01 % (w / v) to about 0.8 % (w / v), about 0.01 % (w / v) to about 0.6 % (w / v), about 0.01 % (w / v) to about 0.4 % (w / v), about 0.01 % (w / v) to about 0.2 % (w / v), about 0.02 % (w / v) to about 5 % (w / v), about 0.02 % (w / v) to about 4 % (w / v), about 0.02 % (w / v) to about 3 % (w / v), about 0.02 % (w / v) to about 2 % (w / v), about 0.02 % (w / v) to about 1 % (w / v), about 0.02 % (w / v) to about 0.8 % (w / v), about 0.02 % (w / v) to about 0.6 % (w / v), about 0.02 % (w / v) to about 0.4 % (w / v), about 0.02 % (w / v) to about 0.2 % (w / v), about 0.03 % (w / v) to about 5 % (w / v), about 0.03 % (w / v) to about 4 % (w / v), about 0.03 % (w / v) to about 3 % (w / v), about 0.03 % (w / v) to about 2 % (w / v), about 0.03 % (w / v) to about 1 % (w / v), about 0.03 % (w / v) to about 0.8 % (w / v), about 0.03 % (w / v) to about 0.6 % (w / v), about 0.03 % (w / v) to about 0.4 % (w / v), about 0.03 % (w / v) to about 0.2 % (w / v), about 0.04 % (w / v) to about 5 % (w / v), about 0.04 % (w / v) to about 4 % (w / v), about 0.04 % (w / v) to about 3 % (w / v), about 0.04 % (w / v) to about 2 % (w / v), about 0.04 % (w / v) to about 1 % (w / v),17IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WO about 0.04 % (w / v) to about 0.8 % (w / v), about 0.04 % (w / v) to about 0.6 % (w / v), about 0.04 % (w / v) to about 0.4 % (w / v), about 0.04 % (w / v) to about 0.2 % (w / v), about 0.05 % (w / v) to about 5 % (w / v), about 0.05 % (w / v) to about 4 % (w / v), about 0.05 % (w / v) to about 3 % (w / v), about 0.05 % (w / v) to about 2 % (w / v), about 0.05 % (w / v) to about 1 % (w / v), about 0.05 % (w / v) to about 0.8 % (w / v), about 0.05 % (w / v) to about 0.6 % (w / v), about 0.05 % (w / v) to about 0.4 % (w / v), about 0.05 % (w / v) to about 0.2 % (w / v), about 0.1 % (w / v) to about 5 % (w / v), or about 0.1 % (w / v) to about 1 % (w / v).

[0093] In some embodiments, the concentration of polysorbate in the pharmaceutical composition is about 0.01 % (w / v), about 0.02 % (w / v), about 0.03 % (w / v), about 0.04 % (w / v), about 0.05 % (w / v), about 0.06 % (w / v), about 0.07 % (w / v), about 0.08 % (w / v), about 0.09 % (w / v), about 0.1 % (w / v), about 0.2 % (w / v), about 0.3 % (w / v), about 0.4 % (w / v), about 0.5 % (w / v), about 0.6 % (w / v), about 0.7 % (w / v), about 0.8 % (w / v), about 0.9 % (w / v), about 1 % (w / v), about 2 % (w / v), about 3 % (w / v), about 4 % (w / v), about 5 % (w / v), about 6 % (w / v), about 7 % (w / v), about 8 % (w / v), about 9 % (w / v), about 10 % (w / v), about 15 % (w / v), about 20 % (w / v), about 25 % (w / v), about 30 % (w / v), about 35 % (w / v), about 40 % (w / v), about 45 % (w / v), or about 50 % (w / v). In some embodiments, the concentration of polysorbate in the pharmaceutical composition is about 2 % (w / v).

[0094] In some aspects, the formulation has a pH of about 2 to about 11. In other aspects, the formulation has a pH of about 4 to about 9.

[0095] In various embodiments, the pH of the pharmaceutical compositions described herein can be from about 5.5 to about 8.5, from about 6 to about 8.5, from about 6.5 to about 8.5, from about 7 to about 8.5, from about 7.5 to about 8.5, from about 8 to about 8.5, from about 5.5 to about 8, from about 5.5 to about 7.5, from about 5.5 to about 7, from about 5.5 to about 6.5, from about 5.5 to about 6, from about 6 to about 8, from about 6 to about 7.5, from about 6 to about 7, from about 6 to about 6.5, from about 6.5 to about 8, from about 6.5 to about 7.5, from about 6.5 to about 7, from about 7 to about 8, from about 7 to about 7.5, or from about 7.5 to about 8. In some embodiments, the pH of the pharmaceutical composition is from about 6.5 to about 8.5. In some embodiments, the pH of the pharmaceutical composition is from about 7 to about 8. In some embodiments, the pH of the pharmaceutical composition is from about 5.5 to about 6.5.

[0096] In various embodiments, the pH of the pharmaceutical compositions described herein can be about 5.5, about 5.9, about 6, about 6.5, about 7, about 7.5, about 8, or about 8.5. In some embodiments, the pH of the pharmaceutical composition is about 5.9.18IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WO

[0097] In various embodiments, the pH of the pharmaceutical compositions described herein can be 5.5±0.1, 6±0.1, 6.5±0.1, 7±0.1, 7.5±0.1, 8±0.1, or 8.5±0.1. In certain embodiments, the pH of the pharmaceutical composition is about 5.9±0.1.

[0098] One aspect of the invention provides a pharmaceutical composition comprising dupilumab and arginine -glutamate. In some embodiments, the pharmaceutical composition further comprises proline. Another aspect of the invention provides a pharmaceutical composition comprising dupilumab, polysorbate, proline, and arginine -glutamate. Another aspect of the invention provides a pharmaceutical composition comprising dupilumab, polysorbate, sucrose, and arginine -glutamate. Another aspect of the invention provides a pharmaceutical composition comprising about 175 mg / mL dupilumab, about 0.2 % (w / v) polysorbate, about 150 mM proline, and about 50 mM arginine-glutamate. Another aspect of the invention provides a pharmaceutical composition comprising about 175 mg / mL dupilumab, about 0.2 % (w / v) polysorbate, about 150 mM sucrose, and about 50 mM arginine-glutamate. In some embodiments, the pharmaceutical composition further comprises polysorbate. In some embodiments, the pharmaceutical composition has a pH of about 5.9. In some embodiments, the pharmaceutical composition is a liquid. In some embodiments, dupilumab has a concentration of about 100 mg / mL to about 200 mg / mL. In some embodiments, dupilumab has a concentration of about 175 mg / mL. In some embodiments, arginine-glutamate has a concentration of about 50 mM. In some embodiments, proline has a concentration of about 150 mM. In some embodiments, polysorbate has a concentration of about 2 mg / mL. In some embodiments, polysorbate has a concentration of about 0.2 % (w / v). In some embodiments, the pharmaceutical composition is substantially free of histidine. In some embodiments, the pharmaceutical composition is substantially free of acetate.

[0099] In some embodiments, the pharmaceutical composition has a viscosity of about 15 to 17 cP. In some embodiments, the pharmaceutical composition has a Tmi of about 66 °C to 68 °C and a Tm2 of about 72 °C to 73 °C. In some embodiments, the pharmaceutical composition has an osmolality of about 300 to 350 mOsm / kg. In some embodiments, the pharmaceutical composition has a polydispersity index (PDI) of about 0. 17 to 0.19. In some embodiments, the pharmaceutical composition has a hydrodynamic radius (Rh) of about 9.5 nm.Methods of Treatment[000100] In one aspect, the pharmaceutical compositions described herein may be used for the treatment or prevention of a variety of diseases and disorders such as, but not limited to, atopic dermatitis, asthma, eosinophilic esophagitis, prurigo nodularis, and chronic rhinosinusitis in a patient in need thereof.19IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WO[000101] In various embodiments, the method comprises administering to the patient a pharmaceutical composition described herein, wherein the pharmaceutical composition generally comprises dupilumab and arginine -glutamate. In some embodiments, the pharmaceutical composition further comprises proline. In some embodiments, the pharmaceutical composition further comprises polysorbate. In some embodiments, the pharmaceutical composition has a pH of about 5.9. In some embodiments, the pharmaceutical composition is a liquid. In some embodiments, dupilumab has a concentration of about 100 mg / mL to about 200 mg / mL. In some embodiments, dupilumab has a concentration of about 175 mg / mL. In some embodiments, arginine-glutamate has a concentration of about 50 mM. In some embodiments, proline has a concentration of about 150 mM. In some embodiments, polysorbate has a concentration of about 2 mg / mL. In some embodiments, the pharmaceutical composition is substantially free of histidine. In some embodiments, the pharmaceutical composition is substantially free of acetate.[000102] In some embodiments, the method comprises administering to the patient a pharmaceutical composition comprising dupilumab and arginine-glutamate. In some embodiments, the pharmaceutical composition further comprises sucrose. In some embodiments, the pharmaceutical composition further comprises polysorbate. In some embodiments, the pharmaceutical composition has a pH of about 5.9. In some embodiments, the pharmaceutical composition is a liquid. In some embodiments, dupilumab has a concentration of about 100 mg / mL to about 200 mg / mL. In some embodiments, dupilumab has a concentration of about 175 mg / mL. In some embodiments, arginine-glutamate has a concentration of about 50 mM. In some embodiments, proline has a concentration of about 150 mM. In some embodiments, polysorbate has a concentration of about 2 mg / mL. In some embodiments, the pharmaceutical composition is substantially free of histidine. In some embodiments, the pharmaceutical composition is substantially free of acetate.[000103] In some embodiments, the method comprises administering to the patient a pharmaceutical composition comprising dupilumab, polysorbate, proline, and arginine-glutamate. In some embodiments, the pharmaceutical composition has a pH of about 5.9. In some embodiments, the pharmaceutical composition is a liquid. In some embodiments, dupilumab has a concentration of about 100 mg / mL to about 200 mg / mL. In some embodiments, dupilumab has a concentration of about 175 mg / mL. In some embodiments, arginine-glutamate has a concentration of about 50 mM. In some embodiments, proline has a concentration of about 150 mM. In some embodiments, polysorbate has a concentration of about 2 mg / mL. In some embodiments, the pharmaceutical composition is substantially free of histidine. In some embodiments, the pharmaceutical composition is substantially free of acetate.20IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WO[000104] In some embodiments, the method comprises administering to the patient a pharmaceutical composition comprising about 175 mg / mL dupilumab, about 2 mg / mL polysorbate, about 150 mM proline, and about 50 mM arginine-glutamate. In some embodiments, the pharmaceutical composition has a pH of about 5.9. In some embodiments, the pharmaceutical composition is a liquid. In some embodiments, the pharmaceutical composition is substantially free of histidine. In some embodiments, the pharmaceutical composition is substantially free of acetate. [000105] In some embodiments, the method comprises administering to the patient a pharmaceutical composition comprising about 175 mg / mL dupilumab, about 2 mg / mL polysorbate, about 150 mM sucrose, and about 50 mM arginine-glutamate. In some embodiments, the pharmaceutical composition has a pH of about 5.9. In some embodiments, the pharmaceutical composition is a liquid. In some embodiments, the pharmaceutical composition is substantially free of histidine. In some embodiments, the pharmaceutical composition is substantially free of acetate. [000106] In some embodiments, the pharmaceutical compositions can be administered parenterally, including by infusion, injection or implantation, which includes subcutaneous administration as appropriate. For example, the pharmaceutical compositions can be administered by, for example, subcutaneous injection or delivery, or intravenous injection or delivery. In some embodiments, the pharmaceutical composition is administered to the patient intravenously. In some embodiments, the pharmaceutical composition is administered to the patient by subcutaneous injection or subcutaneous infusion. In some embodiments, the pharmaceutical composition is administered to the patient by subcutaneous infusion using an on-body, subcutaneous delivery system. In some embodiments, the pharmaceutical composition is administered to the patient by subcutaneous infusion using a wearable subcutaneous delivery system. In some embodiments, the pharmaceutical composition is administered to the patient by subcutaneous infusion using a pump device. In some embodiments, the pump device is a micropump device or a patch device. In some embodiments, the pump device is a patch device.[000107] When administered for the treatment or prevention of a disease or disorder disclosed herein, it may be understood that an effective dosage of a pharmaceutical composition described herein can vary depending upon many factors such as the mode of administration and severity of the condition being treated, as well as the various physical factors related to the individual being treated. In therapeutic applications, a pharmaceutical composition described herein may be provided to a patient already suffering from said disease or disorder in an amount sufficient to cure or at least partially ameliorate the symptoms of the disease or disorder and its complications. The dosage to be used in the treatment of a specific individual typically must be subjectively determined by an attending physician. The variables involved include the specific condition and21IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WO state as well as the size, age and response pattern of the patient. In some embodiments, the amount sufficient to cure or at least partially ameliorate the symptoms of the disease or disorder and its complications is an effective amount. In some embodiments, the amount sufficient to cure or at least partially ameliorate the symptoms of the disease or disorder and its complications is a therapeutically effective amount.EXAMPLES[000108] The invention now being generally described, will be more readily understood by reference to the following examples, which are merely for the purposes of illustration of certain aspects and embodiments of the present invention, and are not intended to limit the invention.AbbreviationsA280 Absorbance at 280 nm A350 Absorbance at 350 nm DLS Dynamic Light Scattering DOE Design of Experiment DP Drag Product DS Drag Substance GMP Good Manufacturing Practices HMW High Molecular Weight icIEF Imaged Capillary Isoelectric Focusing IP Intellectual Property LMW Low Molecular WeightMWCO Molecular Weight Cut Off NT Not Tested NR Non-reducing PDI Polydispersity Index PK Pharmacokinetics Rh Hydrodynamic radius SD Standard Deviation SE-UPLC Size Exclusion Ultra Performance LiquidChromatographyTaggr Aggregation Onset Temperature uv Ultraviolet FT Freeze-thawPS80 Polysorbate 80 TZ Time Zero RH Relative Humidity RP Reference Product UF / DF Ultrafiltration / Diafiltration22IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WOExample 1: pH and buffer concentration investigations[000109] Initial experiments were conducted to investigate different dupilumab formulations. Solutions of dupilumab at approximately 12 mg / mL were tested in six buffer systems of varying pH and buffer concentration. The buffer systems tested were histidineacetate, glutamate, aspartate, citrate, phosphate, and succinate. Nine formulations for each buffer system tested, as shown in Table 1. In addition, 3 buffer-free formulations (pH 5.4, 5.9, and 6.4) were also prepared.Table 1: pH and buffer concentrations for each buffer system tested* Prepared and tested in duplicate to serve as a center point.Preparation of the Formulations[000110] A stock solution with a nominal concentration of 200 mM was prepared for the citrate, aspartate, glutamate, and succinate buffer systems. For histidine-acetate buffer, a stock with nominal concentrations of 200 mM histidine and 125 mM acetate was prepared. For phosphate buffer, a stock solution of 200 mM monobasic sodium phosphate and 200 mM dibasic sodium phosphate were prepared.[000111] For citrate, aspartate, glutamate, and succinate buffer systems, a working solution was prepared by diluting each 200-mM stock solution with water to a nominal concentration of 160 mM and adjusting the pH to the target pH of 5.4, 5.9, and 6.4. For histidine-acetate buffer, the concentrations for the working solution are 160 mM histidine and 100 mM acetate and the pH adjusted to the target pHs. The concentration of the working solution for23IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WO phosphate was 200 mM and was prepared by mixing 200 mM sodium phosphate dibasic and 200 mM sodium phosphate monobasic stock solution at different ratios to achieve the target pHs. The buffer stock and working solutions for each buffer system are summarized in Table 2.Table 2The DS stock solution (85.3 mg / mL in deionized water) and the respective buffer working solution were used in the preparation of the final formulations as shown in Table 3 and Table 4. Further, the three “Buffer-free” formulation were prepared by combining 1.055 mb of the DS stock solution with 4.945 mL of water and adjusting the pH to the target pH of 5.4, 5.9, and 6.4 using 10 mM HC1 or 10 mM NaOH stock solutions.Table 3: Preparation of formulations in histidine-acetate, citrate, aspartate, glutamate, and succinate buffer systems24IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WOTable 4: Preparation of formulations in phosphate buffer system[000112] The final formulations were filtered using Millipore 0.2 pm Steriflip filtration units, manually filled in 2.5 mL aliquots into 2R vials, stoppered with 13 mm serum stopper and sealed with an aluminum flip-off seal by crimping. Finished vials were stored at 2-8 °C prior to testing.Stability Testing[000113] Initial experiments were conducted to investigate the physicochemical stability of the dupilumab formulations. The formulations of dupilumab were stored for 1 month at either 2-8 °C in a refrigerator (control samples) or 40 °C in stability chamber (stressed samples). After 1 month, vials were inspected for visible particulate matter, and aliquoted for analysis.Analysis and Results[000114] Protein concentration Protein concentrations of the stability samples were determined by UV / vis spectroscopy. The UV-Vis absorption spectrum for each sample in the range of 230 nm to 750 nm with 1 nm steps was acquired (Unchained Labs Stunner). Protein concentrations were calculated using an extinction coefficient of 1.41 L g'1cm'1. The protein concentration for control samples were between 12.1 mg / mL and 12.6 mg / mL. Concentrations ranging from 12.2 mg / mL to 13.7 mg / mL were observed for stressed samples, after one month.25IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WO[000115] Aggregation index The aggregation index (Al) was used as a preliminary measure to assess levels of soluble and insoluble aggregates. UV-Vis data (above) was used to calculate aggregation index (Al) for each sample as follows:Al = 100The Al varied from 0.2 to 2.5 for control samples, and from 0.1 to 1.7 for stressed samples. With a few exceptions the Al had increased under 40 °C conditions.[000116] Results of protein concentration and aggregation index measurements are summarized in Figure 1.[000117] pH The pH for each sample (a 400-pL aliquot) was measured using a micro pH electrode. The pH of majority formulations were within the 0.1-0.2 pH units of the initial pH.[000118] Dynamic Light Scattering DLS thermal studies were performed to determine average hydrodynamic radius (Rh), aggregation temperature (Taggr), and polydispersity index (PDI). Studies were performed using a DynaPro Plate Reader II equipped with an 830 nm laser source (Wyatt Technology, Santa Barbara, CA) and Dynamics (Version 7.9. 1.4) data acquisition and processing software. A temperature-dependent Rh measurement was performed in a temperature range of 45 - 80 °C which was used for determining the aggregation temperature (Taggr). Each sample was analyzed in duplicate. The average hydrodynamic radius (Rh) was between 5.5 nm and 6.8 nm for control samples and between 5.6 nm and 7.3 nm for stressed samples after 1 month. The polydispersity index (PDI) was between 0.01 and 0.23 for control samples and between 0.01 and 0.19 for stressed samples after 1 month, showing predominantly single-size species in solution with and without stress. Results of hydrodynamic radius and polydispersity index measurements are summarized in Figure 2.[000119] The thermal stability of dupilumab in all formulations was assessed by the aggregation onset temperature (Taggr). The data are summarized in Figure 3 for control and stressed samples. Higher Taggrusually suggests higher stability of the molecule in formulation.26IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WO[000120] Size exclusion chromatography The size variants of dupilumab (%Monomer,%HMW, and %LMW) present in control and stressed samples were assessed using size exclusion chromatography (SEC). SEC was performed on a Waters ACQUITY IT- Class UPLC system (Waters Corp., Milford, MA). Samples were injected without dilution. A summary of chromatographic conditions is outlined in Table 5. Three peak areas were quantified as a monomer (Main), aggregates (HMW- total of dimer and higher order aggregates), and fragments (LMW).Table 5: SE-UPLC Conditions[000121] Results are summarized in Figure 4. For control samples, the %Main was between 98.4% and 99.6%, with the average above 99.2% in all buffer systems. The %HMW was between 0.36% to 1.42% and the %LMW was between 0.06% and 0.18%. For stressed samples after one month, the amounts of aggregates (%HMW) and fragments (%LMW) increased in all samples. In general, more aggregates were observed in phosphate, succinate, and buffer-free systems than in histidine-acetate, citrate, aspartate, and glutamate buffers. The %LMW, however, was lower in all buffer systems or buffer-free solutions as compared to histidine-acetate buffer.[000122] icIEF Charge variants of dupilumab (%Main peak, %Basic isoforms, and %Acidic isoforms) were assessed by Imaged capillary isoelectric focusing (icIEF) for control and for stressed samples. Analysis was performed on a Maurice system (ProteinSimple / Bio- Techne, Minneapolis, MN). A master mix was prepared according to Table 6. Each sample was diluted by mixing 180 pL of the master mix with 8 pL of sample and 12 pL of water to achieve a final (nominal) sample concentration of 0.5 mg / mL. The diluted samples were27IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WO centrifuged for 3 min at 1000 g. The top 160 pL of each sample was transferred into a Maurice 96-well plate. The sample analysis was performed using Compass software and separation conditions of 1 min at 1500 V, 2-18 min at 3000 V. The sample platform temperature was 10 °C. Data was exported to Empower 2 CDS for peak integration using a predefined processing method with auto integration.Table 6: Master Mix composition[000123] Results are summarized in Figure 5. For stressed samples after one month the main charge variant has decreased, whereas acidic and basic charge variants have increased. The %main for stressed samples was similar in aspartate, citrate, glutamate, and histidineacetate buffers, and somewhat lower in phosphate, succinate, and buffer-free systems. The %basic for stressed samples was similar in all buffer and buffer-free systems, whereas %acidic was lower in aspartate buffer as compared to all other formulations.[000124] Pairwise correlations of %main, %acidic, and %basic charge variants is shown in Figure 6. Strong correlation of %main and %acidic suggests that at 40 °C the main protein isoform is primarily isomerizing into acidic species. Correlation of %main and %basic was not significant.[000125] Optimized pH Analytical responses as a function of measured pH were assessed for hydrodynamic radius (Rh) and polydispersity index (PDI) by DLS, aggregation index (Al) by UV vis spectrophotometry, %HMW and %LMW by SEC, %main, %acidic, and %basic, and Taggr by DLS. Optimal pH ranges for these attributes are summarized in Table 7. More compact structures with lower Rh and PDI are observed at pH below 5.8 or above 6.2, however lower aggregation indexes are observed in the pH range from 5.5 to 6.3. The minimum of %HMW is observed at pH 5.8±0.2, and the minimum of %LMW at pH 6.2±0.2, suggesting that pH around 6.0 is optimal to reduce aggregation and fragmentation. The increase in charge variants after 1 month at 40 °C was lower in samples with pH from28IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WO5.6 to 6.2, with the optimal pH around 6.0, similar to the optimal pH for aggregation and fragmentation. The optimal pH for Taggr is around 6.0, similar to aggregation / fragmentation and to charge isomerization. Based on SEC, icIEF, and DLS data, the optimal formulation pH is 6.0 + / - 0.2.Table 7[000126] Based on the analytical results (Rh by DLS, PDI by DLS, Al, %HMW by SEC, %LMW by SEC, %main by icIEF, %acidic by icIEF, %basic by icIEF and Taggr ) of the buffer systems (histidine-acetate, glutamate, aspartate, citrate, phosphate, succinate and buffer free) after 1 month storage at 40 °C , phosphate, and succinate were eliminated as possible buffers for use in dupilumab formulations. Citrate was also eliminated because it is known to cause painful sensations after subcutaneous injections.Example 2: Viscosity reducer and stabilizer investigationsAdditional experiments were conducted to investigate dupilumab formulations with two classes of excipients: ionic excipients as viscosity reducers (arginine hydrochloride, arginine-glutamate, arginine aspartate, and lysine hydrochloride) and non-ionic excipients as protein stabilizers (sucrose, trehalose, sorbitol, glycine, and proline). Twenty-one formulations were tested (as summarized in Table 8), where each formulation was prepared so that dupilumab had a target concentration of 175 mg / mL, the ionic excipient had a target concentration of 50 mM, the stabilizer29IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WO had a target concentration of 150 mM, and polysorbate 80 had a target concentration of 2 mg / mL.All formulations had a target pH of 5.9.Table 8: Ionic excipients and stabilizers formulations[000127] Each formulation was fdtered using Millipore 0.22 pm Steriflip fdtration units, manually filled in 3.5 m aliquots into 5-mL glass vials, stoppered with 13 mm serum stopper and sealed with an aluminum flip-off seal by crimping. Finished vials were stored at 2-8 °C until setting up normal stability and forced degradation studies.Stability Studies[000128] For stability studies, vials were stored for 1 month at 2-8 °C in a refrigerator(control samples, also referred to as TZ in the figures) or at 40 °C in a stability chamber (stressed) for 1 month.[000129] The freeze-thaw stress (FT) samples were frozen at -20 °C for ~22 h and thawed at 2-8 °C for ~23 h. Three total freeze-thaw cycles were performed for each sample. Samples30IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WO were then stored at 2-8 °C in a refrigerator for 1 month. After one month, all samples (control (also referred to as TZ in the figures), stressed, and FT) were pulled, inspected for visible particulate matter and aliquoted for analysis.Analysis and Results[000130] Visual Inspection All samples were free from visible particles when analyzed against white and black background using the visual inspection booth.[000131] Protein concentration and Al The protein concentration and the aggregation index (Al) of the formulations was determined by UV / vis spectroscopy by the same method as reported in Example 1. The protein concentration for all samples was within the target+ 10%, except 193.8 mg / mL for ArgAspSuc forthe control sample (TZ)s and 198.3 mg / mL for LysHClSuc after FT stress. These results show that protein concentration was consistent, with no significant change after 1 month at 40 °C or after three freeze-thaw cycles. The Al for all control samples was between 0.024 and 0.089, after three freeze-thaw cycles the Al was between 0.032 and 0.073, and after 1 month at 40 °C the Al was between 0.147 and 0.202. No significant change in Al was observed after the freeze-thaw stress, however, after 40 °C stress the Al significantly increased in all formulations. The increase of Al after 40 °C storage can be attributed to the formation of soluble and / or insoluble aggregates. Results of protein concentration measurements are summarized in Figure 7. Results of protein concentration and aggregation index measurements are summarized in Figure 8.[000132] pH The pH of each sample was measured by the same method as reported in Example 1.[000133] Dynamic Light Scattering DLS thermal studies were performed to determine average hydrodynamic radius (Rh) and polydispersity index (PDI) of each sample as reported in Example 1. The Rh for control samples was from 9.8 nm to 15.9 nm. After three freeze-thaw cycles the Rh was between 9.8 nm and 15.7 nm, with no significant change as compared to control samples for all formulations. After 1 month at 40 °C the Rh was between 10.2 nm and 16.9 nm, an increase in all formulations as compared to control samples. The PDI ranges were from 0.158 to 0.40 for control samples, from 0.163 to 0.335 after three freeze-thaw cycles, and from 0.189 to 0.386 after 1-month storage at 40 °C. In general, after 40 °C stress the PDI increased as compared to control samples. A PDI31IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WO significantly higher than 0.2 indicates size heterogeneity. The results of the DLS experiments are reported in Figure 9.[000134] The diffusion interaction parameter (ko) for each formulation was determined by measuring the diffusion coefficient (D) as a function of protein concentration (c) using Stunner (Unchained Labs, Pleasanton, CA). Each sample was diluted with matching formulation buffers to reach protein concentrations in the range of 1 to 25 mg / mL. Diffusion interaction parameters were obtained by the equation:Oc=DQ(1 + kDx c) where Dcis the mutual diffusion coefficient, Do is the diffusion coefficient near zero concentration, ko is diffusion interaction parameter, and c is protein concentration. A linear plot of c vs Dcgives a slope and intercept of koDo and Do, from which ko can be calculated using / cD=slope / intercept. Diffusion coefficients were measured for each formulation in the range from 5 mg / mL to 25 mg / mL. The calculated ko values were all negative, ranging from -4.4 to -10.1 mL / g, which is indicative of protein-protein attraction in all formulations. This behavior is typical for antibodies of IgG4 subclass. The values for ko are summarized in Figure 10. Based on the results from these experiments, lysine was excluded as possible excipients in further formulation development.[000135] Size exclusion chromatography The size variants of dupilumab (%Monomer, %HMW, and %LMW) present in the formulations were assessed using size exclusion chromatography (SEC). SEC was performed on a Waters ACQUITY H-Class UPLC system (Waters Corp., Milford, MA). The formulations were diluted using their respective buffers to a nominal concentration of 20 mg / mL. A summary of chromatographic conditions is outlined in Table 5, above, except the injection volume was 1.5 pL. The results of the SEC measurements are reported in Figure 11. The %Monomer was between 97.5% and 98.0% for control samples, between 97.5% and 98.0% for FT samples, and between 95.5% and 96.8% for 40 °C stressed samples. No significant change was detected for %Monomer after three freeze-thaw cycles in all formulations. The %Monomer decreased by l.l%-2.2% after 1-month storage at 40 °C with glycine-containing formulations showing the largest decrease in %Monomer. The %HMW was between 1.8% and 2.3% for control samples, between 1.9% and 2.3% for FT samples, and between 3.0% and 4.2% for 40 °C stressed samples. No significant change was detected for %HMW after three freeze-thaw cycles in all formulations. The %HMW increased by 1.0%-2.0% after 1-month storage at 4032IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WO°C. The %LMW was 0.2% for control samples, 0.2% for FT samples, and 0.3% for 40 °C stressed samples. No significant change was detected for %LMW after three freeze-thaw cycles, and only minor increase by 0. 1% after 1-month storage at 40 °C was detected in all formulations. The increase in %HMW was observed in all formulations after 1 month at 40 °C, however, more aggregation was detected in glycine -containing formulations. Based on the results of the experiment, glycine was excluded as a possible excipient in further formulation development.[000136] icIEF Charge variants of dupilumab (%Main peak, %Basic isoforms, and %Acidic isoforms) were assessed by imaged capillary isoelectric focusing (icIEF) for the formulations by the same method as reported in Example 1. Each formulation was diluted by mixing 180 pL of the master mix (see Table 6 above) with 20 pL of 5-mg / mL sample (diluted by water) to achieve a final (nominal) sample concentration of 0.5 mg / mL. The %Main isoform was between 67.6% and 81.4% for control samples, between 73.3% and 85.3% for FT samples, and between 46.9% and 59.8% for 40 °C stressed samples. No change larger than 5% was observed for %Main after three freeze-thaw cycles as compared to control samples for the majority of formulations with an exception of ArgAspPro, ArgGluGly, ArgGluSor, and ArgGluSuc formulations, for which the %Main increased by more than 5%. After 1 month at 40 °C the %Main decreased from 16.8% to 28.4% depending on formulation. The % Acidic isoform was between 15.8% and 23.5% for control samples, between 13.1% and 24.7% for FT samples, and between 32.6% and 42.4% for 40 °C stressed samples. No change larger than 5% was observed for %Acidic after three freeze-thaw cycles as compared to control samples for the majority of formulations with an exception of ArgAspPro, ArgAspSor, and ArgGluSor, for which the %Acidic decreased by more than 5%. After 1 month at 40 °C the %Acidic increased from 13.7% to 21.3% depending on formulation. The 18.3% increase %Acidic was detected in the reference formulation ArgHCl+HisAc. The % Basic isoform was between 2.7% and 9.3% for control samples, between 1.5% and 4.4% for FT samples, and between 2.8% and 11.4% for 40 °C stressed samples. No change larger than 2% was observed for %Basic after three freeze-thaw cycles as compared to control samples for the majority of formulations, with an exception of ArgAspSuc and ArgGluSorb, for which the %Basic decreased by more than 2%. After 1 month at 40 °C the %Basic increased from nearly 0% to 7.7% depending on formulation. Charge isomerization of the protein takes place in all formulations after 1 month at 40° C,33IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WO with main isoform transitioning into acidic and basic isoforms. The results of the icIEF measurements are reported in Figure 12.[000137] Differential Scanning Calorimetry The protein unfolding heat capacity in each formulation as a function of temperature was assessed using differential scanning calorimetry (DSC). The temperatures of unfolding onset (Tonset), and domain melting temperatures (Tmi, Tm2, and Tm-i) were obtained. The DSC measurements were performed on a MicroCai VP-Capillary DSC with autosampler (Malvern Panalytical, Westborough, MA). MicroCai VP-Capillary DSC software 4.0 was used for data collection. MicroCai VP- Capillary DSC Automated Analysis software was used for data analysis. All samples were diluted to a nominal protein concentration of 1 mg / mL using the respective formulation buffer. The results are reported in Figure 13.[000138] Viscosity This viscosity (cP) for each formulation was measured on a VROC initium viscometer (RheoSense, San Ramon, CA) at 20 °C. Solution viscosity as a function of protein concentration was evaluated in 50 mM arginine hydrochloride at pH 5.9. Each sample was measured in 10 replicates. The viscosity values reported are the averages of 9 replicates excluding the first replicate. The viscosities for the formulations are shown in Figure 14. The solution viscosities at 20 °C vary from 14 cP to 25 cP. In addition to formulation composition, the actual protein concentration variability can introduce changes into solution viscosity, particularly due to very high protein concentration. In order to compare the formulations, the viscosity at 175 mg / mL was calculated using the following linear approximation ni75=tactual — 0.4(Cactual -175) where nns and Cactuai are measured viscosities and protein concentrations. The calculated viscosities of the formulations at 175 mg / mL (p 175) is shown in Figure 15. The solution viscosities at 175 mg / mL do not exceed a set threshold amount, except formulations ArgAspGly, ArgAspTre, ArgHClTre, LysHClSor, LysHClSuc, and LysHClTre. Based on the viscosity experiments, these formulations could potentially be more challenging and may be excluded as possible excipients in further formulation development.34IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WOResults: Statistical analysis[000139] A full-factorial design with two categorical factors of four levels (ionic excipients / viscosity reducers, including ArgHCl, ArgGlu, ArgAsp, and LysHCl) and five levels (stabilizers, including sucrose, trehalose, sorbitol, glycine, and proline) was implemented. The analytical data collected for the 21 formulations (20 formulations from the full-factorial design with the reference formulation) were analyzed using JMP.[000140] Two excipients, ionic excipient lysine hydrochloride and stabilizer lysine had the largest number of “worst” responses compared to the other excipients. For lysine hydrochloride, these critical responses were higher solution viscosity and more negative ko value, suggesting higher risk of potential aggregation. For glycine, aggregation attributes (%Monomer 40 °C, %HMW 40 °C) and fragmentation were the most critical attributes. As a result, lysine hydrochloride and glycine were excluded from further formulation development.[000141] Excipients arginine hydrochloride, arginine-glutamate, arginine aspartate, trehalose, sorbitol, and proline were found to be compatible with the dupilumab drug substance based on the results from the JMP analysis. However, due to other issues with the excipients (weak buffers, Tg’, solubility concerns, etc.), arginine aspartate, sorbitol, and trehalose were excluded from further formulation development. Proline was shown to have superior viscosity reducing properties. Arginine-glutamate was found to be superior in comparison to arginine hydrochloride based on Rh by DLS and higher thermal stability by DSC.[000142] Based on the analytical and statistical results, arginine hydrochloride, arginine- glutamate, sucrose, and proline were selected for further formulation development.Example 3. Formulation Development: Comparative Stability Study of Stable Dupilumab Pharmaceutical Formulations[000143] Following the investigations summarized in Examples 1 and 2, three formulations and a reference formulation were prepared at 175 mg / mL nominal concentration as summarized in Table 9. Samples were filled into 2.25 mb pre-filled syringes and subjected to stress conditions including freeze-thaw cycles, mechanical shaking, and photo-stress. Stability studies at 2-8 °C, 25 °C, and 40 °C for up to 6 months were also conducted. Formulations were assessed for chemical, colloidal, conformational, and interfacial stability relative to the reference formulation as a control. Solution viscosity was also studied for assessing manufacturability.35IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WOTable 9: Formulations Tested3Alias refers to the abbreviated formulation ID used during sample analysis.Stability Schedule and Testing[000144] The stability samples for the formulations and the reference formulation were set down at 2-8 °C, 25 °C, and 40 °C and samples were pulled after 0, 1, 3, and 6 months according to Table 10.Table 10Freeze-thaw[000145] The formulations were subjected to 5 freeze-thaw cycles in which each cycle consisted of freezing at -20 °C overnight and thawing at 2-8 °C overnight. All FT stressed samples were stored at 2-8 °C before testing.Photo stress[000146] The photo stress for the formulations was performed in a photostability chamber (Environmental Specialties, Raleigh NC). The samples (in PFS) received 0.2 times of ICH recommended light exposure (0.24 million lux hours white light and 40 watt hours / square meter near UV light). A separate set of the formulation samples wrapped in aluminum foil was placed under the same photo-stress condition serving as negative controls. All photo-stressed samples were stored at 2-8 °C before testing.36IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WOShaking stress[000147] The formulations were wrapped in aluminum foil and shaken on a VWR DVX-2500 Multi-Tube Vortexer at a speed setting of 500 rpm for 48 hours at room temperature. A separate set of the formulation samples wrapped in aluminum foil was placed at room temperature serving as negative controls. After the shaking stress, the samples were stored at 2-8 °C prior to the testing.Analysis and Results[000148] Visual Inspection All formulations were essentially free from visible particles when visually inspected against white and black background on a visual inspection booth.[000149] pH The pH of each formulation was measured by the method described in Example 1. The pH values were in the range of 5.9-6. 1. No significant shift in pH was observed among the stability and stress samples for all formulations.[000150] Protein concentration Protein concentration was measured by the method disclosed in Example 1. The protein concentration for all formulations were in the range of 169 - 181 mg / mL. The results are reported in Figure 16. There were no significant changes in protein concentration among the stability samples and the stressed samples.[000151] Aggregation Index Aggregation Index (Al) was measured by the method disclosed in Example 1. The results are reported in Figure 17. There was no significant increase in Al for the formulations, except samples stored at 40°C, for which Al increase was observed for all samples.[000152] Dynamic Light Scattering DLS was measured by the method disclosed in Example 1. Polydispersity index (PDI) and hydrodynamic radius (Rh) for the formulations, as well as the reference formulation, were measured at 175 mg / mL protein concentration by DLS without sample dilution. A value over 0.2-0.3 is generally considered as more heterogeneous in size. The PDI values for ArgGlu Proline formulation samples are in general lower than other formulations, see Figure 18, indicating more size homogeneity in the ArgGlu Proline formulation.37IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WO[000153] Size exclusion chromatography The size exclusion chromatography experiments were conducted by the method described in Example 2. No significant change in %HMW were observed in any of the lead formulations stored at 2-8 °C for 6 months, see Figure 19 and Figure 20B. A slight increase of %HMW2 was observed for ArgGlu formulations stored at 25 °C for 6 months, see Figure 20C. The overall change in %HMW for all formulations at 25 °C, however, follows similar trend, see Figure 19, increased from 2.3-2.5% at time 0 to 2.9-3.2% at 6 months. A higher-order aggregate species (HMW3) is observed for all formulations at 40 °C, see Figure 20D, which is similar to the aggregation behavior of the reference formulation under the same condition. There were no significant differences on the aggregation kinetics for lead formulations at 2-8 °C, 25 °C, and 40 °C, see Figures 19 and Figure 21. A slight upward trend is observed for all formulations when stressed at 40 °C for 3 months, consistent with increased aggregation levels as detected by SEC, see Figure 19. No significant changes in aggregation was observed after freeze-thaw and shaking stress, see Figure 22. The overall %HMW increased significantly for photo stressed lead formulation samples as shown in Figure 22, at 3.9-4.6% as compared to 2.9% in the reference formulation, which indicates lower photo stability for all three lead formulations. The SEC data suggests that the lead formulations are more sensitive to light stress than the reference formulation. There are no significant differences between the lead formulations and the reference formulation in the real time, accelerated, and stressed stability, as well as freeze-thaw and mechanical shaking stress.[000154] icIEF The icIEF experiments were conducted by the methods described in Example 1. Typical electropherograms for the formulations are shown in Figure 23, together with the electropherograms of the reference formulation as a comparison. The overall charge variants profile for the formulations are similar to that of the reference formulation. There is no significant difference between the lead formulations and the reference formulation after 6 months at 25 °C. The stability trend for %Main and %Acidic charge variants for all lead formulations at 2-8 °C, 25 °C, and 40 °C are similar to that of the reference formulation, see Figure 24 and Figure 25. When the No Buffer formulation was stored at 40 °C for 3 months, an additional acidic peak with a pl of -7.27, see Figure 26 is observed, which is not in the reference formulation. Further, the level of the basic charge variant at pl - 7.47 is increased to 9.7% in the No Buffer formulation, as compared to 6.5% for the reference formulation, see Figure 26 and 27.[000155] No significant changes in the charge heterogeneity were observed in ArgGlu- Sucrose and ArgGlu-Proline formulations, which suggests these two formulations help to maintain similar level of chemical stability. No Buffer formulation offers less protection against chemical38IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WO degradation, as evidenced by the elevated chemical degradation level when stressed for 3 months at 40 °C.[000156] CE-SDS The CE-SDS analysis was performed on a Maurice system (ProteinSimple / Bio-Techne, Minneapolis, MN). The formulation samples were diluted to a nominal concentration of 10 mg / mL. For non-reducing (NR) CE-SDS analysis, 5 pL of the diluted 10-mg / mL sample was mixed with 45 pL of IX CE-SDS Plus sample buffer, 2 pL of 25X internal standard, and 2.5 pL of 250 mM lodoacetamide. For reducing (R) CE-SDS analysis, 5 pL of the diluted 10-mg / mL sample was mixed with 45 pL of IX CE-SDS Plus sample buffer, 2 pL of 25X internal standard, and 2.5 pL of 2-mercaptoethanol. The samples (NR and R) were loaded onto a 96-well PCR plate, heated at 70°C for 10 min, then cooled on ice for 5 min, and centrifuged at 3700 rpm for 10 min. The samples were analyzed using a Turbo CE-SDS cartridge with instrument parameters as listed in Table 11.Table 11.The example electropherograms for the formulations at time 0 and 6 months at 25 °C were shown in Figure 28. There is no significant difference observed for the formulations at Time 0 and 6 months at 25 °C. The stability trend of the protein purity (%IgG) for lead formulations and the reference formulation at 2-8 °C, 25 °C, and 40 °C are shown in Figure 29. The stability trend for the lead formulations at all three temperatures are comparable to the reference formulation. The purity of the protein (%IgG) in lead formulations are in the range of 91.3-94.0%, as compared to 95.1% in the reference formulation Figure 29. The purity dropped to the range of 94.2-94.7% for the formulations stored at 25 °C for 6 months. The overall NRCE-SDS data indicate there is no significant difference in the size heterogeneity between the lead formulations and the reference formulation.[000157] The reduced CE-SDS electropherograms for the formulations showed similar size variant profile as the reference formulation at time 0 and after 6 months at 25 °C, see Figure 30. There are no discernible differences in the purity (%HC+%LC) between lead formulations and the reference formulation at 2-8 °C for up to 6 months. A slight decrease of the purity is observed at 25 °C, from 98.8-99.5% at time 0 to 98.0-98.5% at 6 months across all formulations, with the same stability trends. Similar stability trends are also observed at 40 °C. There is no change between39IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WO lead formulations and the reference formulation in the stressed samples. There were no significant differences in size heterogeneity in the stability and stressed samples for the lead formulations and the reference formulation as analyzed by RCE-SDS.[000158] Differential Scanning Calorimetry DSC measurements were obtained by the methods described in Example 2. Replacing histidine-acetate buffer system with ArgGlu buffer system resulted in an increase of Tm, see Figure 31, which indicates higher conformational stability of the formulations ArgGlu-Sucrose and ArgGlu Proline relative to the reference formulation.[000159] Viscosity Viscosity of the samples were measured by the methods described inExample 2. The protein concentration dependent viscosity profile for the formulations were measured and used to evaluate the manufacturability of each lead formulation Figure 32. The viscosity for all formulations at 175 mg / mL is approximately 15-17 cP, and approximately 10 cP at 150 mg / mL. The viscosity for the two lead formulations, No Buffer and ArgGlu-Sucrose, are estimated to be 40 and 49 cP at 210 mg / mL, respectively, which are not optimal for large-scale UF / DF operation. With an expected viscosity of 31 cP at 210 mg / mL, which is lower than that of the reference formulation (34 cP), ArgGlu-Proline is advantageous in terms of manufacturability and injectability.[000160] Osmolality The osmolality of the formulations were measured on an OsmoPro Osmometer (Advanced Instruments, Norwood, MA). The osmolality for all lead formulations at 175 mg / mL are within the range of 311-346 mOsm / kg.[000161] Based on the analytical results, ArgGlu-Proline formulation was selected as the formulation for dupilumab due to its lower viscosity, comparable chemical, colloidal, and interfacial stability, and a slightly improved conformational stability as compared to the reference formulation.[000162] The formulation nominated is 50 mM arginine-glutamate, 150 mM proline, 0.2% polysorbate 80, with a pH of 5.9. This formulation is suitable for dupilumab at both 150 mg / mL and 175 mg / mL.40IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WOEQUIVALENTS[000163] The disclosure may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. The foregoing embodiments are therefore to be considered in all respects illustrative rather than limiting the disclosure described herein. Scope of the disclosure is thus indicated by the appended claims rather than by the foregoing description, and all changes that come within the meaning and range of equivalency of the claims are intended to be embraced therein.41IPTS / 200164313.2 103069-456772

Claims

Attorney Docket No. TPH-001WOCLAIMSWHAT IS CLAIMED:

1. A pharmaceutical composition comprising: dupilumab; and arginine-glutamate .

2. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition further comprises proline.

3. The pharmaceutical composition of claim 1 or 2, wherein the pharmaceutical composition further comprises polysorbate.

4. The pharmaceutical composition of any one of claims 1 to 3, wherein the pharmaceutical composition has a pH of about 5.9.

5. The pharmaceutical composition of any one of claims 1 to 4, wherein the pharmaceutical composition is a liquid.

6. The pharmaceutical composition of any one of claims 1 to 5, wherein dupilumab has a concentration of about 100 mg / mL to about 200 mg / mL.

7. The pharmaceutical composition of any one of claims 1 to 6, wherein dupilumab has a concentration of about 175 mg / mL.

8. The pharmaceutical composition of any one of claims 1 to 7, wherein arginine-glutamate has a concentration of about 50 mM.

9. The pharmaceutical composition of any one of claims 1 to 8, wherein proline has a concentration of about 150 mM.

10. The pharmaceutical composition of any one of claims 1 to 9, wherein polysorbate has a concentration of about 2 mg / mL.

11. The pharmaceutical composition of any one of claims 1 to 9, wherein polysorbate is present in an amount of about 0.2% (w / v).

12. The pharmaceutical composition of any one of claims 1 to 11, wherein the pharmaceutical composition is substantially free of histidine.

13. The pharmaceutical composition of any one of claims 1 to 12, wherein the pharmaceutical composition is substantially free of acetate.

14. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition further comprises sucrose.

15. The pharmaceutical composition of claim 14, wherein the pharmaceutical composition further comprises polysorbate.42IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WO16. The pharmaceutical composition of any one of claims 14 to 15, wherein the pharmaceutical composition has a pH of about 5.9.

17. The pharmaceutical composition of any one of claims 14 to 16, wherein the pharmaceutical composition is a liquid.

18. The pharmaceutical composition of any one of claims 14 to 17, wherein dupilumab has a concentration of about 100 mg / mL to about 200 mg / mL.

19. The pharmaceutical composition of any one of claims 14 to 18, wherein dupilumab has a concentration of about 175 mg / mL.

20. The pharmaceutical composition of any one of claims 14 to 19, wherein arginineglutamate has a concentration of about 50 mM.

21. The pharmaceutical composition of any one of claims 14 to 20, wherein proline has a concentration of about 150 mM.

22. The pharmaceutical composition of any one of claims 14 to 21, wherein polysorbate has a concentration of about 0.2% (w / v).

23. The pharmaceutical composition of any one of claims 14 to 22, wherein the pharmaceutical composition is substantially free of histidine.

24. The pharmaceutical composition of any one of claims 14 to 23, wherein the pharmaceutical composition is substantially free of acetate.

25. A pharmaceutical composition comprising: dupilumab; polysorbate; proline; and arginine-glutamate .

26. The pharmaceutical composition of claim 25, wherein the pharmaceutical composition has a pH of about 5.9.

27. The pharmaceutical composition of claim 25 or 26, wherein the pharmaceutical composition is a liquid.

28. The pharmaceutical composition of any one of claims 25 to 27, wherein dupilumab has a concentration of about 100 mg / mL to about 200 mg / mL.

29. The pharmaceutical composition of any one of claims 25 to 28, wherein dupilumab has a concentration of about 175 mg / mL.

30. The pharmaceutical composition of any one of claims 25 to 29, wherein arginine-glutamate has a concentration of about 50 mM.43IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-001WO31. The pharmaceutical composition of any one of claims 25 to 30, wherein proline has a concentration of about 150 mM.

32. The pharmaceutical composition of any one of claims 25 to 31, wherein polysorbate has a concentration of about 0.2% (w / v).

33. The pharmaceutical composition of any one of claims 25 to 32, wherein the pharmaceutical composition is substantially free of histidine.

34. The pharmaceutical composition of any one of claims 25 to 33, wherein the pharmaceutical composition is substantially free of acetate.

35. A pharmaceutical composition comprising: about 175 mg / mL dupilumab; about 2 mg / mL polysorbate; about 150 mM proline; and about 50 mM arginine-glutamate.

36. The pharmaceutical composition of claim 35, wherein the pharmaceutical composition has a pH of about 5.9.

37. The pharmaceutical composition of claim 35 or 36, wherein the pharmaceutical composition is a liquid.

38. The pharmaceutical composition of any one of claims 35 to 37, wherein the pharmaceutical composition is substantially free of histidine.

39. The pharmaceutical composition of any one of claims 35 to 38, wherein the pharmaceutical composition is substantially free of acetate.

40. A pharmaceutical composition comprising: about 175 mg / mL dupilumab; about 2 mg / mL polysorbate; about 150 mM sucrose; and about 50 mM arginine-glutamate.

41. The pharmaceutical composition of any one of claims 1 to 40, wherein the pharmaceutical composition has a viscosity of about 15 to 17 cP.

42. The pharmaceutical composition of any one of claims 1 to 41, wherein the pharmaceutical composition has a Tmi of about 66 °C to 68 °C and a Tm2 of about 72 °C to 73 °C.

43. The pharmaceutical composition of any one of claims 1 to 42, wherein the pharmaceutical composition has an osmolality of about 300 to 350 mOsm / kg.

44. The pharmaceutical composition of any one of claims 1 to 43, wherein the pharmaceutical composition has a polydispersity index (PDI) of about 0. 17 to 0.19.44IPTS / 200164313.2 103069-456772Attorney Docket No. TPH-OOIWO45. The pharmaceutical composition of any one of claims 1 to 44, wherein the pharmaceutical composition has a hydrodynamic radius (Rh) of about 9.5 nm.45IPTS / 200164313.2 103069-456772

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