High-concentration antibody composition

Abstract

Provided are a method for producing an aqueous antibody composition containing at least 200 mg / ml of an antibody and a method for producing a dried antibody composition. A high-concentration antibody composition is further provided. Further provided is the use of the provided antibody composition in medicine.

Description

Technical Field

[0001] Methods for producing an aqueous antibody composition containing an antibody at a concentration of at least 200 mg / ml and a method for producing a dried antibody composition are provided. A high-concentration antibody composition is further provided. Use of the provided antibody composition in medicine is further provided.

Background Art

[0002] Antibodies are multifunctional components of the immune system. A large number of monoclonal antibodies have been approved as pharmaceuticals in the past few decades. Monoclonal antibodies (or fragments thereof) against different antigens are produced and used in medicine, for example, for the treatment of immune diseases and cancer.

[0003] Combined with the desire for long-term stability, the limited stability of antibodies, especially highly concentrated antibodies, has been a driving force for developing methods capable of stabilizing proteins. Dehydration of protein formulations to yield dry powders and the addition of excipients are the two most common stabilization techniques for extending shelf life. Protein stability can be significantly enhanced by removing water from the formulation. This is a result of the reduced mobility of the protein and the lack of certain degradation pathways facilitated by water. Under appropriate excipients and drying conditions, the protein is incorporated into an amorphous matrix, which is maintained as long as the temperature is maintained below the glass transition temperature, thereby enhancing the long-term stability of the protein. In addition, cryoprotectants such as trehalose and sucrose stabilize the protein by excluding water from the vicinity of the protein, protecting it from degradation during drying, storage of the dry powder, and storage of the aqueous solution when rehydrated. Surfactants are often used to prevent protein adsorption at the interface in order to avoid denaturation at the air-water interface.

[0004] Antibody formulations with good long-term stability and appropriate powder properties are needed. The long-term stability of dry powders depends mainly on the water content (typically less than 5%), formulation design, protein structure, and storage conditions. Drying is often necessary to obtain a product that is stable over the long term, but the drying process can result in a combination of heat, interfacial, and mechanical stresses that can adversely affect protein stability. To mitigate the effects of these stresses on the product, optimized drying process parameters and the addition of excipients are required. The addition of excipients changes the impact of the drying process and makes this process protein- and formulation-specific.

[0005] Several techniques can be used to dry the protein and obtain a protein powder. These include drying techniques such as freeze-drying or lyophilization, spray drying, spray freeze-drying or prilling, and supercritical fluid drying. All of these techniques use different physical principles for drying, induce different stresses during drying, and result in powders with different particle or powder properties.

[0006] Gikanga et al. have disclosed a method for the production of high-concentration monoclonal antibody formulations via spray drying (Gikanga et al., PDA J Pharm Sci Technol. 2015 Jan-Feb;69(1):59-73).

[0007] Deokar et al. have provided a comparison of various techniques for obtaining high-concentration (about 200 mg / mL) antibody formulations (Deokar et al. J Pharm Sci. 2020;109(12):3579-3589).

[0008] Clenet D, Hourquet V, Woinet B, Ponceblanc H, Vangelisti M. A spray freeze dried micropellet based formulation proof-of-concept for a yellow fever vaccine candidate. Eur J Pharm Biopharm. 2019 Sep;142:334-343。

[0009] WO 2018 / 204374 A1 pamphlet discloses an aqueous antibody formulation.

[0010] US 2020 / 390705 A1 specification discloses a spray-dried antibody formulation.

[0011] US 11,351,256 B2 specification, WO 2013 / 016648 A2 pamphlet and WO 2013 / 063510 A1 pamphlet disclose methods for obtaining antibody formulations.

[0012] However, high-concentration pharmaceutical antibody formulations can have many problems, such as stability issues like the formation of visible particles, invisible particles, and antibody aggregates, and solution property issues like high turbidity. It is important that antibody formulations for parenteral use have a low aggregate level (up to a maximum of 5% high molecular weight species or HMWS based on generally accepted industry specifications), small invisible particles (compliant with USP <787> specifications, acceptance level: ≤6000 particles of 10 μm per container, ≤600 particles of 25 μm per container), low turbidity (up to a maximum of 40 NTU based on commercial product benchmarking, Kingsbury JS et al. J Pharm Sci. 2021 Sep;110(9):3176-3182. doi:10.1016 / j.xphs.2021.05.005. Epub 2021 May 15. PMID:34004217), and no visible particles. USP <787> is the standard established by "Subvisible Particulate Matter in Therapeutic Protein Injections" in USP chapter <787> of the United States Pharmacopeia Convention (USP).

[0013] Therefore, it is important to avoid the formation of antibody aggregates (which can be measured by measuring the change in the percentage of high molecular weight species). It is important that the final antibody formulation used by the end user has stability attributes that meet the specifications, and at the same time, to obtain a commercially viable product, the stability of the antibody during drying, during storage of the dried intermediate product (e.g., powder or micropellets) at the recommended storage temperature and shelf life, and during storage as a solution at the recommended storage temperature and shelf life must be ensured.

[0014] In particular, when attempting to administer a high dose of an antibody, for example, via subcutaneous administration using a prefilled syringe, a highly concentrated antibody pharmaceutical formulation, such as a formulation containing at least 200 mg / ml of an antibody, is required. If a formulation containing an antibody at 200 mg / mL or higher can be obtained, it becomes possible to increase the amount of the antibody drug substance in a single prefilled syringe, and thus it becomes possible to administer a higher dose via subcutaneous administration instead of intravenous administration, or in some cases, the number of subcutaneous injections is reduced, and both of these promote patient-centered treatment.

Summary of the Invention

Problems to be Solved by the Invention

[0015] Therefore, there is still a need for improved high-concentration pharmaceutical antibody compositions that avoid these limitations and meet typical acceptance specifications.

Means for Solving the Problems

[0016] A method for preparing an aqueous antibody composition I containing at least 200 mg / ml of an antibody is provided herein, the method comprising steps a), b) and c) a) An aqueous composition A, a1) An antibody at a concentration lower than the concentration of the antibody in antibody composition I, a2) At least one surfactant, a3) At least one cryoprotectant, a4) At least 0.4% (w / v) of arginine or a salt thereof (where the weight (w) is calculated as arginine) based on the total volume of the aqueous composition A, for example, 0.4 - 3.5% (w / v) of arginine An aqueous composition A comprising, wherein the ratio of the amount of a1) to the sum of the amounts of a3) and a4) is 11:1 to 11:8, and optionally, a5) A buffer Providing an aqueous composition A containing b) Drying the aqueous composition A provided in step a) to obtain an antibody composition B, c) Reconstituting the composition dried in step b) to obtain an aqueous antibody composition I comprises.

[0017] In one embodiment of the provided method, the salt of arginine is selected from the group consisting of arginine-Cl, arginine-aspartate, arginine-glutamate, arginine-sulfate, arginine-acetate, arginine-succinate, and mixtures thereof.

[0018] In one embodiment of the provided method, arginine is arginine-Cl.

[0019] In one embodiment of the provided method, at least one cryoprotectant is selected from the group consisting of trehalose, sucrose, sorbitol, glycerol, and mannitol.

[0020] In one embodiment of the provided method, the cryoprotectant is trehalose.

[0021] In one embodiment of the provided method, the ratio of the amount of a1) to the sum of the amounts of a3) and a4) is from 11:1 to 11:4, for example from 11:2 to 11:4.

[0022] In one embodiment of the provided method, the aqueous composition A comprises less than 200 mg / ml of antibody, for example from 10 to 190 mg / ml of antibody, for example from 20 to 170 mg / ml of antibody, for example from 50 to 150 mg / ml of antibody, for example from 80 to 120 mg / ml of antibody.

[0023] In one embodiment of the provided method, the aqueous composition A comprises 150 mg / ml of antibody.

[0024] In one embodiment of the provided method, the aqueous antibody composition I comprises more than 200 mg / ml of antibody.

[0025] In one embodiment of the provided method, the aqueous antibody composition I comprises at least 250 mg / ml of antibody, for example more than 250 mg / ml of antibody.

[0026] In one embodiment of the provided method, aqueous antibody composition I comprises at least 300 mg / ml of antibody, such as an antibody in excess of 300 mg / ml.

[0027] In one embodiment of the provided method, aqueous antibody composition I comprises from 230 mg / ml to 420 mg / ml of antibody, such as from 260 mg / ml to 400 mg / ml of antibody.

[0028] In one embodiment of the provided method, aqueous antibody composition I comprises from 270 to 350 mg / ml of antibody, such as from 280 to 320 mg / ml of antibody, such as 300 mg / ml of antibody.

[0029] In one embodiment of the provided method, the antibody is a monoclonal antibody.

[0030] In one embodiment of the provided method, at least one surfactant is a polysorbate such as polysorbate 20 or 80 or a poloxamer such as poloxamer 188.

[0031] In one embodiment of the provided method, the buffer is a histidine, citrate, acetate, phosphate, Tris, succinate or glycine buffer.

[0032] In one embodiment of the provided method, the pH of composition A is from about 5.0 to 8.0, such as from about 5.5 to 7.

[0033] In one embodiment of the provided method, antibody composition B is in the form of a powder or micro-pellets.

[0034] In one embodiment of the provided method, step b) is carried out by lyophilization or spray drying to give a lyophilized or spray dried powder.

[0035] In one embodiment of the provided method, step b) is carried out by spray-freeze drying to give spray-freeze dried micro-pellets.

[0036] In one embodiment of the provided method, the lyophilized powder is reconstituted in the reconstitution solution over a period of 1.5 to 4.0 hours.

[0037] In one embodiment of the provided method, the spray-dried powder is reconstituted in the reconstitution solution over a period of at least 4 hours.

[0038] In one embodiment of the provided method, the spray-freeze-dried micropellets are reconstituted in the reconstitution solution over a period of 1 to 2 hours.

[0039] In some embodiments, the powder or micropellets are reconstituted in water. Thus, the reconstitution solution can be water.

[0040] In one embodiment of the provided method, the aqueous antibody composition I is a clear and transparent solution.

[0041] There is further provided an aqueous antibody composition obtainable or obtained by the method for producing an aqueous antibody composition provided herein.

[0042] There is further provided an aqueous antibody composition I containing at least 200 mg / ml of an antibody.

[0043] In an embodiment of the provided aqueous antibody composition I, it contains an antibody at 270 to 350 mg / ml, such as an antibody at 280 to 320 mg / ml, such as an antibody at 300 mg / ml.

[0044] In another embodiment of the provided aqueous antibody composition I, it contains an antibody at 230 mg / ml to 420 mg / ml, such as an antibody at 260 mg / ml to 400 mg / ml.

[0045] In one embodiment, the aqueous antibody composition I is a clear and transparent solution.

[0046] In one embodiment, the aqueous antibody composition I contains at least 200 mg / ml of an antibody and has an opalescence of less than 40 NTU, for example, less than 25 NTU or less than 15 NTU.

[0047] In one embodiment, the aqueous antibody composition I further contains at least one surfactant and contains at least one cryoprotectant, such as trehalose and arginine or a salt thereof.

[0048] Also provided herein is a container containing the aqueous antibody composition I provided herein.

[0049] In one embodiment of the provided container, the container is a prefilled syringe, vial, cartridge, ampoule or autoinjector. For example, the container can be an autoinjector or a prefilled syringe.

[0050] Also provided herein is a kit comprising the container provided herein and a label or instructions for administration and use of the aqueous antibody composition I.

[0051] Further provided herein is the aqueous antibody composition provided herein for use in medicine.

[0052] A method for preparing a dry antibody composition B, a) An aqueous composition A, a1) An antibody of less than 200 mg / ml, for example, an antibody of 10 - 190 mg / ml, for example, an antibody of 20 - 170 mg / ml, for example, an antibody of 50 - 150 mg / ml, for example, an antibody of 80 - 120 mg / ml, and a2) At least one surfactant, and a3) At least one cryoprotectant, and a4) At least 0.4% (w / v) of arginine or a salt thereof (wherein the weight (w) is calculated as arginine) based on the total volume of the aqueous composition A, for example, 0.4 - 3.5% (w / v) of arginine Provided is an aqueous composition A, which contains and has a ratio of the amount of a1) to the total amount of a3) and a4) of 11:1 to 11:8, and Also provided is a method of spray freeze-drying the composition provided in step a) to give a dried antibody composition B.

[0053] In one embodiment of the above method, the dried antibody composition B is in the form of micropellets.

[0054] Also provided are a dried antibody composition B obtainable or obtained by a method for producing the dried antibody composition B and a container containing the dried antibody composition B.

[0055] Also provided are a dried antibody composition B and a container containing the dried antibody composition B. The dried antibody composition B is in the form of micropellets, and the composition B a1) an antibody, and a2) at least one surfactant, and a3) at least one cryoprotectant, such as trehalose, and a4) arginine or a salt thereof contains, and the ratio of the amount of a1) to the total amount of a3) and a4) is 11:1 to 11:8, for example 11:2 to 11:8, and the weight of a4) is calculated as the free amino acid.

[0056] Also provided are a dried antibody composition B and a container containing the dried antibody composition B. The dried antibody composition B is in the form of powder, and the composition B a1) an antibody, and a2) at least one surfactant, and a3) at least one cryoprotectant, such as trehalose, and a4) arginine or a salt thereof contains, and the ratio of the amount of a1) to the total amount of a3) and a4) is 11:1 to 11:8, for example 11:2 to 11:8, and the weight of a4) is calculated as the free amino acid.

[0057] Further provided is the use of the dry antibody composition B provided herein for preparing an aqueous antibody composition I as defined herein, comprising at least 200 mg / ml of an antibody.

[0058] Further provided is the use of an aqueous composition A for preparing an aqueous antibody composition I as defined herein, comprising at least 200 mg / ml of an antibody.

[0059] Further provided is an aqueous composition A as defined in relation to the method for preparing an aqueous antibody composition I. The definition is applied according to the situation.

[0060] Therefore, an aqueous composition A, a1) less than 200 mg / ml of an antibody, for example 10 - 190 mg / ml of an antibody, for example 20 - 170 mg / ml of an antibody, for example 50 - 150 mg / ml of an antibody, for example 80 - 120 mg / ml of an antibody, and a2) at least one surfactant, and a3) at least one cryoprotectant, and a4) at least 0.4% (w / v) of arginine or its salt (where the weight (w) is calculated as arginine) based on the total volume of the aqueous composition A, for example 0.4 - 3.5% (w / v) of arginine, and is provided, wherein the ratio of the amount of a1) to the sum of the amounts of a3) and a4) is 11:1 - 11:8.

Brief Description of the Drawings

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DETAILED DESCRIPTION OF THE INVENTION

[0062] Definitions Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. Here, note that when used in this specification and the appended claims, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise. Further, as used herein, the term "at least one" will be understood to mean one or more. For example, 2, 3, 4 or more. Depending on the item, when present, the term refers to what upper limit the term may refer to, as understood by one of ordinary skill in the art.

[0063] As used herein, the term "about" means that there is an interval accuracy that can achieve the technical effect with respect to any number listed after the term. Thus, for example, when referred to herein, "about" refers to the exact numerical value or a range around the exact numerical value of ±20%, ±15%, ±10% or ±5%. In one embodiment, the term refers to the exact value.

[0064] As used herein, the term "comprising" should not be understood in a limiting sense. Rather, this term indicates that there may be more than the actual items mentioned. For example, when referring to a method comprising a particular step, the presence of additional steps is not excluded. However, the term "comprising" also encompasses embodiments in which only the items mentioned are present, i.e., has a limiting meaning in the sense of "consisting of".

[0065] The aqueous composition A provided in step a) of the method of the present invention contains an antibody.

[0066] As used herein, the term "antibody" refers to an immunoglobulin or immunoglobulin-like molecule. This includes, by way of example, IgA, IgD, IgE, IgG and IgM, combinations thereof and any vertebrate, such as mammals such as goats, rabbits and mice and similar molecules produced during an immune response in non-mammalian species, such as shark immunoglobulins, but is not limited thereto. The term includes synthetic antibodies, monoclonal antibodies, recombinantly produced antibodies, multispecific antibodies (including bispecific antibodies), human antibodies, humanized antibodies, single domain antibodies (immunoglobulin single variable domains (ISVDs), also referred to as, for example, Nanobody® molecules), which include single specificity, bispecific, multispecific single domain antibodies, chimeric antibodies, intrabodies, single chain Fv (scFv) (including, for example, single specificity, bispecific, etc.), camelized antibodies, any of the foregoing, Fab fragments, F(ab’)2 fragments, disulfide bond Fv (sdFv), anti-idiotype (anti-Id) antibodies and epitope binding fragments. In particular, an antibody includes an immunoglobulin molecule and an immunologically active portion of the immunoglobulin molecule, i.e., an antigen-binding domain or molecule containing an antigen-binding site that specifically binds to an antigen (e.g., the six complementarity determining regions (CDRs) of an antibody). An antibody can be of any class of immunoglobulin molecule (e.g., IgG, IgE, IgM, IgD, IgA and IgY), any subclass (e.g., IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2) or any sub-subclass (e.g., IgG2a and IgG2b). In some embodiments, the antibody is humanized. In certain embodiments, the antibody is an IgG antibody, such as an IgG1 or IgG4 antibody. In one embodiment, the antibody is an anti-IgG4 antibody.

[0067] In one embodiment, the antibody or fragment thereof is produced recombinantly.

[0068] The light chain contains two domains or regions, a variable domain (VL) and a constant domain (CL). The heavy chain contains four domains, a variable domain (VH) and three constant domains (CH1, CH2, and CH3, collectively referred to as CH). The variable regions of both the light (VL) and heavy (VH) chains determine the binding recognition and specificity for antigens. The constant region domains of the light chain (CL) and heavy chain (CH) confer important biological properties such as antibody chain association, secretion, transplacental mobility, complement binding, and binding to Fc receptors (FcR). The Fv fragment is the N-terminal portion of the Fab fragment of an immunoglobulin and consists of the variable portions of one light chain and one heavy chain. The specificity of an antibody lies in the structural complementarity between the antibody binding site and the antigen determinant. The antibody binding site is mainly composed of residues derived from the hypervariable or complementarity-determining regions (CDRs). Occasionally, residues from non-hypervariable regions or framework regions (FRs) affect the overall domain structure and thus the binding site. The term "complementary determining region" (abbreviated as CDR) refers to the amino acid sequence that together defines the binding affinity and specificity of the native Fv region of the native immunoglobulin binding site. The light and heavy chains of an immunoglobulin each have three CDRs, designated CDR1-L, CDR2-L, CDR3-L (in the case of light chain complementarity-determining regions) or CDRL1, CDRL2, CDRL3 and CDR1-H, CDR2-H, CDR3-H (in the case of heavy chain complementarity-determining regions) or CDRH1, CDRH2, CDRH3, respectively. Thus, a conventional antibody antigen binding site contains six CDRs, including a set of CDRs from each of the heavy and light chain variable regions.

[0069] In one embodiment, the antibody is a monoclonal antibody. As used herein, the term "monoclonal antibody" refers to an antibody produced by a single clone of B lymphocytes or an antibody produced by a cell into which the light chain gene and heavy chain gene of a single antibody have been transfected. Monoclonal antibodies are produced by methods known to those skilled in the art, such as by creating hybrid antibody-producing cells from the fusion of myeloma cells and immune spleen cells.

[0070] In one embodiment, the antigen-binding protein is a diabody. A diabody is a bivalent antibody comprising two polypeptide chains. Each polypeptide chain comprises a variable heavy chain domain and a variable light chain domain linked by a linker. In one embodiment, the two polypeptide chains of the diabody are identical. In an alternative embodiment, the two polypeptide chains have different amino acid sequences, provided that the two chains bind to the same antigen (either the same epitope or different epitopes within the same antigen).

[0071] The term "antibody" can also include a mixture of antibodies, such as a mixture of two or more antibodies.

[0072] In one embodiment, the antibody is a therapeutic antibody, i.e., an antibody used in therapy. For example, the antibody is selected from the group consisting of adalimumab, belimumab, atezolizumab, elotuzumab, infliximab, evolocumab, nivolumab, cetuximab, pembrolizumab, durvalumab, siltuximab, eculizumab, fremabnezumab-vfrm, ofatumumab, omalizumab, galcanezumab-gnlm, rituximab, panitumumab, daratumumab, palivizumab, denosumab, mepolizumab, golimumab, trastuzumab, obinutuzumab, abemaciclib, ocrelizumab, pertuzumab, vedolizumab, ramucirumab, tocilizumab, secukinumab, orlatumumab, necitumumab, guselkumab, ustekinumab, bevacizumab, alemtuzumab, certolizumab (pegol), and benralizumab.

[0073] In addition to the antibody, the "composition" or "formulation" optionally contains additional components in amounts as specified herein. Excipients refer to inert substances commonly used as diluents, vehicles, preservatives, binders, stabilizers, etc. for drugs, including, but not limited to, amino acids (e.g., arginine), fatty acids and phospholipids, surfactants (e.g., polysorbate, nonionic surfactants, etc.), sugars (e.g., sucrose, maltose, trehalose, etc.). "Pharmaceutically acceptable excipient" means any inert substance combined with an antibody as referred to herein to prepare a preferred or convenient dosage form. A "pharmaceutically acceptable excipient" is an excipient that is non-toxic to the recipient at the dosages and concentrations used and is compatible with the other components of the formulation containing the monoclonal antibody.

[0074] a1) In addition to the antibody, Composition A further comprises a2) at least one surfactant, a3) at least one cryoprotectant, and a4) arginine or a salt thereof. Additionally, the composition may contain A5) a buffer. Typically, the composition contains excipients in amounts as specified elsewhere herein.

[0075] Surfactants are chemical compounds that interact with and stabilize biomolecules and / or common pharmaceutical excipients in formulations. Surfactants generally protect binders from air / solution interface-induced stress and solution / surface-induced stress, which otherwise could cause protein aggregation. Examples of surfactants include, but are not limited to, polysorbates, glycerin, dicarboxylic acids, oxalic acid, succinic acid, fumaric acid, phthalic acid, and combinations thereof. Those skilled in the art know that other surfactants, such as nonionic or ionic detergents, can be used as long as they are pharmaceutically acceptable, i.e., suitable for administration to a subject. In some embodiments, the surfactant is a polysorbate. Examples of polysorbates include polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 65, and polysorbate 80. In one embodiment, the polysorbate is polysorbate 80. In one embodiment, the polysorbate is polysorbate 20. Polysorbate 80, polysorbate 20, and poloxamer 188 are the most commonly used surfactants in commercially available parenteral products.

[0076] In some embodiments, the surfactant is a poloxamer, such as poloxamer 188.

[0077] Typically, the surfactant is present in Composition A in an amount of about 0.001% to about 0.2% (w / v) based on the total volume of Composition A. For example, the surfactant can be present in Composition A in amounts of about 0.005% (w / v), about 0.006% (w / v), about 0.007% (w / v), about 0.008% (w / v), about 0.009% (w / v), 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) and about 0.2% (w / v). In some embodiments, the surfactant is present in Formulation A in an amount of about 0.01% to about 0.1% (w / v), about 0.02% to about 0.08% (w / v) or about 0.03% to about 0.07% (w / v), such as about 0.05% (w / v). For example, polysorbate 80 can be present in an amount of about 0.05% (w / v).

[0078] Since the surfactant is present in Composition A, it is also present in Composition I.

[0079] A cryoprotectant is a chemical compound that protects the antibody contained in the composition from freeze damage, i.e., damage caused by ice formation or adsorption to the ice - liquid interface. Thus, the cryoprotectant provides stability to the antibody in the formulation and alleviates the stress induced by freezing. The cryoprotectant can be a saccharide such as trehalose, sucrose, glucose, mannitol, mannose and lactose or a polymer such as polyethylene glycol. Typically, the cryoprotectant is selected from the group consisting of cryoprotectants consisting of trehalose, sucrose, sorbitol, glycerol and mannitol.

[0080] In some embodiments, the cryoprotectant is trehalose.

[0081] The formulations or aqueous compositions as referred to in this specification contain the amino acid arginine or a salt thereof. Typically, the composition contains a salt of arginine. In one embodiment, the arginine salt is selected from the group consisting of arginine-Cl (also called arginine-HCl), arginine-aspartate, arginine-glutamate, arginine-sulfate, arginine-acetate, arginine-succinate, and mixtures thereof.

[0082] In one embodiment, the salt is arginine-Cl.

[0083] The aqueous composition may contain a5) a buffering agent. The buffering agent is an agent that maintains a physiologically suitable pH. In addition, the buffering agent may enhance the isotonicity and chemical stability of the formulation. For example, the buffering agent may be selected from histidine, citric acid, acetic acid, phosphoric acid, tris, succinic acid, or glycine buffer.

[0084] In some embodiments, the buffering agent is present in Composition A at a concentration of about 0.5 mM to about 50 mM, such as about 5 mM to about 15 mM. For example, it may be present at a concentration of about 15 mM.

[0085] In certain embodiments, the composition provided in step a) of the method provided herein may have a pH in the range of about 5.0 to about 8.0, such as about 5.5 to about 7.0. For example, the pH of Composition A can be about 5.5, about 5.7, about 5.9, about 6.1, about 6.1, about 6.3, about 6.5, about 6.7, about 6.9, and about 7.0. In some embodiments, the pH of Composition A can be in the range of about 5.8 to about 6.3. The pH of Composition A can be measured by any means known to those skilled in the art. The means for measuring pH is to use a pH meter. The pH of Composition A can be adjusted using any means known in the art. The chemicals for changing the pH of Composition A are hydrochloric acid (HCl) and sodium hydroxide (NaOH).

[0086] In some embodiments, the pH of Composition A is antibody-specific, i.e., selected to provide optimal stability. For example, the pH can be below or above the isoelectric point (pI) of the antibody. The isoelectric point is the pH at which a particular molecule or surface bears no net charge. In one embodiment, the pH value of Composition A is not the isoelectric point of the antibody. For example, the pH value of Composition A can be at least 0.5 above or below the isoelectric point of the antibody.

[0087] In step b) of the method of the present invention, Composition A as provided in step a) is dried to obtain Antibody Composition B.

[0088] As used herein, the terms "drying" or "dehydrating" refer to removing water from the aqueous Composition A, thereby producing Composition B. Typically, the water content (and thus the moisture content) of Composition B is less than 10%, such as less than 5%, such as less than 3%. In one embodiment, the water content is from 0.5% to 5%, such as from 0.5% to 3%. The water content or moisture content is typically the amount of water contained in Composition B, measured by Karl-Fischer moisture analysis. By carefully selecting the drying process parameters, a lower moisture content can be achieved. A lower moisture content will ensure better stability during storage of Composition B. In the research underlying the present invention, Karl Fischer was used to determine the residual water content. The oven temperature was set at 120 °C, the air flow rate was set at 80 mL / min, and the drift had to be less than 20 μg water / min to start the measurement. The stop criterion was having a relative drift around 15 μg water / min. As the anode solution in the titration cell, Hydranal coulomat AG-oven (Honeywell Fluka) reagent was used. Prior to the test sample, a 1% water standard (MilliporeSigma) was run to confirm that the apparatus was operating correctly. In one embodiment, the moisture content as referred to herein is measured by this method.

[0089] Drying can be achieved by any method considered appropriate, for example, by techniques such as freeze-drying (or lyophilization), spray-drying, and spray-freeze-drying. Typically, drying is achieved by freeze-drying, spray-drying, or spray-freeze-drying.

[0090] In one embodiment, the drying step is performed by freeze-drying (often referred to as "lyophilization").

[0091] As used herein, the term "freeze-drying" (or "lyophilization" or "cryodesiccation") refers to a method in which Composition A is cooled to a temperature at which the water in the composition freezes. The frozen water is then removed by one or two drying steps, a primary drying step (including removal of unbound water by sublimation) and optionally a subsequent secondary drying step (including removal of bound solvents by desorption).

[0092] In another embodiment, the drying step is carried out by spray-drying.

[0093] Spray-drying is a three-step process that includes removal of the aqueous solvent by evaporation following a spraying step and subsequent powder recovery. The protein feed solution is sprayed directly into the drying chamber where the droplets come into contact with a hot drying gas such as air or nitrogen. The drying time is very short and can vary from seconds to minutes depending on the scale of the spray dryer. After drying, the powder is recovered by either a cyclone or a filter. Process parameters such as feed flow rate, drying air flow rate, spray flow rate, and inlet temperature, and formulation parameters such as excipients and solid concentration affect the moisture content, particle size, particle morphology, density, and protein stability. Mechanical stress during spraying, thermal stress during drying, and interfacial stress throughout the process can adversely affect the stability of the product. An increase in the drying temperature reduces the moisture content, which helps protein stability but at the same time results in more thermal stress and potential denaturation.

[0094] In another embodiment, the drying step is carried out by spray-freeze-drying.

[0095] Spray freeze drying (often also referred to as "prilling") is a four-step process that includes a spraying step, spray freezing the droplets, thereby "confining" them in a spherical droplet shape, subsequently performing freeze drying, and then recovering the micropellets (see, for example, WO 2013 / 050156 A1 pamphlet or WO 2009 / 109550 A1 pamphlet). The physical and morphological properties of the resulting powder are mainly affected by the first two steps, while the third step relates to the drying time and the total energy required to dry the product. Process parameters such as feed flow rate, nozzle configuration, freezing rate, freezing temperature, drying air flow rate and pressure, and formulation parameters such as excipients and solid concentration affect the water content, particle size, particle morphology, density and protein stability. The mechanical stress during spraying, the stress associated with ice during freezing, the thermal stress during drying, and the interfacial stress throughout the process can adversely affect the stability of the product. This technology enables highly controlled residual moisture content, mass density and particle size. Examples of spray freeze drying are disclosed in the Examples section.

[0096] The composition B obtained in step b) can be in the form of a powder (i.e., dry powder) or micropellets. For example, the micropellets can be normal spherical micropellets or particles having a diameter of about 400 μm to about 700 μm. The diameter can be measured by a scanning electron microscope (SEM).

[0097] The form of the composition B can depend on the drying method. In one embodiment, step b) is carried out by freeze drying or spray drying to give a (freeze dried or spray dried) powder. In another embodiment, step b) is carried out by spray freeze drying to give spray freeze dried micropellets.

[0098] In one embodiment, the composition B is reconstituted to obtain an aqueous antibody composition I.

[0099] In step c) of the method of the present invention, the dry composition B is reconstituted to obtain an aqueous antibody composition I, i.e., a composition containing at least 200 mg / ml of antibody.

[0100] Step c) can be carried out immediately after the composition B is obtained. In another embodiment, the composition B is stored for a certain period of time before step c) is carried out.

[0101] The reconstitution of the composition B can be carried out in any aqueous (reconstitution) solution (e.g., an aqueous buffer) considered to be appropriate.

[0102] In one embodiment, the reconstitution solution is an aqueous solution containing at least one additive selected from the group consisting of water, glycine, arginine-Cl, guanidinium-Cl, arginine-glutamate, and magnesium glutamate.

[0103] An aqueous antibody composition I or composition B as referred to in this specification is a "stable" composition. A stable composition is one in which the antibody essentially retains its physical stability, identity, integrity and / or chemical stability, identity, integrity and / or biological activity during storage. Various analytical techniques for measuring protein stability are available in the art. Stability can be measured at selected temperatures and other storage conditions over a selected period. Stability can be determined by examining the aggregation or level of high molecular weight species using size exclusion chromatography. For example, an antibody "retains its physical stability" in a composition if visual inspection of color and / or transparency and characterization of the level of high molecular weight species shows no significant signs of increased aggregation level, precipitation and / or denaturation. In some embodiments, when using the compositions of the invention, when measured by size exclusion chromatography (e.g., as described in the Examples section) or any other suitable method for measuring aggregate formation, the antibody forms aggregates of 5% or less, typically 4% or less, typically 3% or less, more typically 2% or less and particularly 1% or less. For example, an antibody is considered stable in a particular composition if the antibody monomer has a purity of about 90% or more, typically about 95% or more or about 98% or more after a particular predetermined period under particular storage conditions in a particular formulation. In one embodiment, less than 5% of the antibody forms aggregates. Chemical stability can be evaluated by detecting and quantifying the chemically altered forms of the protein. Chemical changes can include, for example, size modifications (e.g., clipping) that can be evaluated using (HP)SEC, SDS-PAGE and / or matrix-assisted laser desorption ionization / time-of-flight mass spectrometry (MALDI / TOF MS). Other types of chemical changes include, for example, changes in charge that can be evaluated by ion exchange chromatography.

[0104] The term "opalescence" (often also called "turbidity") refers to an optical property of a solution caused by the scattering of light, which can be explained by different scattering events such as Rayleigh scattering or Mie scattering. The opalescence of a solution can be evaluated as described, for example, in Kingsbury JS et al.. J Pharm Sci. 2021 Sep;110(9):3176-3182. doi:10.1016 / j.xphs.2021.05.005. Epub 2021 May 15. PMID:34004217 (which is hereby incorporated by reference in its entirety). In one embodiment, Composition A and aqueous antibody composition I as referred to herein have an opalescence of less than 40 Nephelometric Turbidity Units (NTU), for example less than 25 NTU. For example, the opalescence can be less than 15 NTU. In one embodiment, the dry antibody composition A as referred to herein has an opalescence of less than 40 Nephelometric Turbidity Units (NTU), for example less than 25 NTU. For example, the opalescence can be less than 15 NTU. In one embodiment, the aqueous antibody composition I as referred to herein has an opalescence of less than 40 Nephelometric Turbidity Units (NTU), for example less than 25 NTU. For example, the opalescence can be less than 15 NTU.

[0105] The aqueous antibody composition I or Composition B as referred to can be used in therapy. Accordingly, the composition can be administered to a subject in need thereof, i.e., a subject suffering from a disease or disorder or at risk of suffering from a disease or condition.

[0106] When Composition B is used in therapy, it is necessary to reconstitute the composition to obtain the aqueous antibody composition I as described elsewhere herein.

[0107] In one embodiment, the term "disease or disorder" refers to any pathological or unhealthy condition that can be treated by administering the formulations or compositions provided herein.

[0108] The "subject" or "patient" can be a vertebrate. The term "subject" includes both humans and other animals, particularly mammals and other organisms. Thus, as used herein, a subject can be an animal such as a mouse, rat, hamster, rabbit, guinea pig, ferret, cat, dog, chicken, sheep, bovine species, horse, camel, or primate. In some embodiments, the subject is a mammal. In some embodiments, the subject is a primate. In some embodiments, the subject is a human.

[0109] The term "administer" or "administration" refers to the act of injecting or otherwise physically delivering a substance (e.g., a formulation of the invention) to a patient when the substance is present outside the body, such as by mucosal, intradermal, intravenous, subcutaneous, intramuscular delivery, and / or any other physical delivery method described herein or known in the art. In one embodiment, the composition is administered subcutaneously. In another embodiment, the composition is administered intravenously.

[0110] The compositions described herein are typically administered in a therapeutically effective amount. The term "therapeutically effective amount" is understood by those skilled in the art. In some embodiments, the term optionally refers to an amount that achieves a desired therapeutic response or desired therapeutic effect, alone or in combination with additional dosages, without or only minimally causing unacceptable or undesirable side effects.

[0111] As used herein, the term "administering" or "administration" refers to the administration of a compound or composition or combination of compounds or compositions for the following purposes: preventing, alleviating or eliminating in a subject a disease and / or disorder as referred to herein, such as obesity. Thus, the term encompasses the treatment of an existing disease or disorder as referred to herein or the prevention of a disease or disorder, i.e., both prevention. Thus, it will be appreciated that in some embodiments, the treatment as referred to herein may be prophylactic. In some embodiments, the term refers to the treatment of an existing disease or disorder as referred to herein. Thus, the subject is suffering from said disease or disorder.

[0112] Provided herein is a method for preparing an aqueous antibody composition I comprising at least 200 mg / ml of an antibody, said method comprising steps a), b) and c) a) an aqueous composition A, a1) an antibody at a concentration lower than the concentration of the antibody in the antibody composition I, a2) at least one surfactant, a3) at least one cryoprotectant, a4) at least 0.4% (w / v) arginine or a salt thereof (wherein the weight (w) is calculated as arginine), for example 0.4 - 3.5% (w / v) arginine comprising, and providing an aqueous composition A wherein the ratio of the amount of a1) to the sum of the amounts of a3) and a4) is from 11:1 to 11:8; b) drying the composition provided in step a) to obtain an antibody composition B; c) reconstituting the composition dried in step b) to give an aqueous antibody composition I and comprising.

[0113] The aqueous antibody composition I produced by the method of the present invention comprises a high concentration of an antibody, such as a monoclonal antibody, typically a therapeutic monoclonal antibody.

[0114] Typically, aqueous antibody composition I contains at least 200 mg / ml of an antibody, such as more than 200 mg / ml of an antibody.

[0115] Typically, aqueous antibody composition I also contains at least 250 mg / ml of an antibody, such as at least 300 mg / ml of an antibody.

[0116] In one embodiment, aqueous antibody composition I contains an antibody at a concentration of 230 mg / ml to 420 mg / ml, such as 260 mg / ml to 400 mg / ml.

[0117] In one embodiment, aqueous antibody composition I contains an antibody at a concentration of 270 to 350 mg / ml, such as 280 to 320 mg / ml.

[0118] In one embodiment, aqueous antibody composition I contains 300 mg / ml of an antibody.

[0119] Typically, aqueous antibody composition I is a clear and transparent solution. This can be evaluated visually (by the appearance of the composition).

[0120] The provided method for preparing aqueous antibody composition I is a) providing an aqueous composition A; b) drying the composition provided in step a) to obtain an antibody composition B; c) reconstituting the composition dried in step b) to give an aqueous antibody composition I and includes.

[0121] The aqueous composition provided in step a) is a1) an antibody at a concentration lower than the concentration of the antibody in antibody composition I; a2) at least one surfactant; a3) at least one cryoprotectant; a4) at least 0.4% (w / v) arginine or a salt thereof (where the weight (w) is calculated as arginine), such as 0.4 to 3.5% (w / v) arginine including, the ratio of the amount of a1) to the sum of the amounts of a3) and a4) is from 11:1 to 11:8, and the aqueous composition optionally a5) a buffering agent is included.

[0122] The concentration of the antibody in a1) is lower than the concentration of the antibody in the aqueous antibody composition I. For example, when the concentration of the antibody in composition I is at least 200 mg / ml, the concentration of said antibody in composition A is less than 200 mg / ml. In some embodiments, the concentration of the antibody in composition A is at least 5%, such as at least 10%, such as at least 20%, such as at least 20% lower than the concentration of the antibody in composition I. For example, the concentration of the antibody in composition A is 10 to 70%, such as 30% to 60% lower than the concentration in composition I.

[0123] In one embodiment, the aqueous composition A contains less than 200 mg / ml of the antibody.

[0124] In one embodiment, the aqueous composition A contains 10 to 190 mg / ml of the antibody.

[0125] In one embodiment, the aqueous composition A contains 20 to 170 mg / ml of the antibody.

[0126] In one embodiment, the aqueous composition A contains 50 to 150 mg / ml of the antibody.

[0127] In one embodiment, the aqueous composition A contains 80 to 120 mg / ml of the antibody.

[0128] In one embodiment, the aqueous composition A contains 100 mg / ml of the antibody.

[0129] The term "surfactant" is defined above. Further, typical concentrations of the surfactant are given above. In one embodiment, at least the surfactant is a polysorbate such as polysorbate 80. In another embodiment, the surfactant is a poloxamer such as poloxamer 188.

[0130] The term "antifreeze agent" is defined above. For example, the antifreeze agent is selected from the group of antifreeze agents consisting of trehalose, sucrose, sorbitol, glycerol and mannitol. In one embodiment, the antifreeze agent is trehalose.

[0131] The aqueous composition A may contain a buffer as described above in a5).

[0132] In one embodiment, the buffer is a histidine buffer.

[0133] In an alternative embodiment, the buffer is a citrate buffer.

[0134] In an alternative embodiment, the buffer is an acetate buffer.

[0135] In an alternative embodiment, the buffer is a phosphate buffer.

[0136] In an alternative embodiment, the buffer is a Tris buffer.

[0137] In an alternative embodiment, the buffer is a succinate buffer.

[0138] In an alternative embodiment, the buffer is a glycine buffer.

[0139] Furthermore, in one embodiment, the pH value of the composition is about 5.0 to 8.0, for example about 5.5 to 7 (however, it may depend on the antibody).

[0140] Typically, the aqueous composition A contains at least 0.4% (w / v) arginine or a salt thereof, calculated as arginine, based on the total volume of composition A. For example, the aqueous composition A contains 0.4 to 3.5% (w / v) arginine (as the free base). In one embodiment, the aqueous composition A contains 1.0 to 3.0% (w / w) arginine (as the free base), for example 1.25 to 2.8% (w / w) arginine, etc.).

[0141] When aqueous composition A contains an arginine salt, it should be understood that the amount is also calculated as the amount of free arginine base present. The free arginine base has a molecular weight of 174 g / mol. The term "amount" typically refers to weight.

[0142] In one embodiment, the ratio of the amount of a1) to the sum of the amounts of a3) and a4) is from 11:1 to 11:8. For example, this ratio can be about 11:1, about 11:2, about 11:3, about 11:4, about 11:5, about 11:6, about 11:7, about 11:8. The ratio of the amount of a1), i.e., the amount of the antibody, to the sum of the amounts of a3) and a4), i.e., the amount of the excipient, should be understood as a weight / weight (w / w) ratio.

[0143] In one embodiment, the ratio of the amount of a1) to the sum of the amounts of a3) and a4) is from 11:1 to 11:4.

[0144] In one embodiment, the ratio of the amount of a1) to the sum of the amounts of a3) and a4) is from 11:2 to 11:4.

[0145] In one embodiment, the ratio of the amount of a1) to the sum of the amounts of a3) and a4) is from 11:2 to 11:3.

[0146] In one embodiment, the ratio of the amount of a1) to the sum of the amounts of a3) and a4) is from 11:1 to 11:4.

[0147] In some embodiments, a4) is a salt of arginine. In one embodiment, it has been shown that the stability of the antibody during the drying process is enhanced by the presence of at least 0.4% arginine in composition A (see Examples).

[0148] In one embodiment, the arginine salt is arginine-Cl (arginine-HCl, see also the Examples section).

[0149] In an alternative embodiment, the arginine salt is arginine-aspartate.

[0150] In an alternative embodiment, the arginine salt is arginine-glutamate.

[0151] In an alternative embodiment, the arginine salt is arginine-sulfate.

[0152] In an alternative embodiment, the arginine salt is arginine-acetate.

[0153] In an alternative embodiment, the arginine salt is arginine-succinate.

[0154] In step b) of the method of the present invention, the aqueous composition A is dried to obtain the antibody composition B. After drying, the water content of the dried composition, i.e., composition B, is less than 10%, for example less than 5%, for example less than 3%.

[0155] In one embodiment, the composition A is dried by lyophilization. Lyophilization typically results in a powder. Thus, the composition B is provided in the form of a powder.

[0156] In an alternative embodiment, the composition A is dried by spray drying. Spray drying typically results in a powder. Thus, the composition B is provided in the form of a powder.

[0157] In an alternative embodiment, the composition A is dried by spray freeze-drying. Spray freeze-drying typically results in a powder. Thus, the composition B is provided in the form of micro pellets.

[0158] In step c) of the method of the present invention, the dried composition B is reconstituted to give an aqueous antibody composition I, i.e., a composition containing at least 200 mg / ml of antibody.

[0159] Step c) can be carried out immediately after Composition B is obtained. However, since Composition B has been shown in the research underlying the present invention to have long-term stability, it is also contemplated that Composition B be stored for a certain period (see Examples). For example, Composition B can be stored for at least 1 month, for example at least 3 months, for example at least 6 months. In one embodiment, the present composition is stored for 1 to 3 months. In an alternative embodiment, Composition B is stored for 3 to 12 months, for example 3 to 6 months. Composition B can be stored at a temperature of 2°C to 25°C, for example 5°C to 25°C. Typically, Composition B is stored refrigerated at a temperature of 2°C to 8°C or stored at a temperature of 17°C to 23°C under ambient conditions.

[0160] The reconstitution of Composition B can be carried out in any aqueous (reconstitution) solution (e.g., an aqueous buffer) that is considered appropriate. In the studies described in the Examples section, Composition B was reconstituted without problems in the following aqueous reconstitution solutions: · 1.5% or 200 mM glycine, · 3% or 150 mM arginine-Cl, · 6.4% or 200 mM arginine-glutamate, · 5.8% or 150 mM magnesium glutamate, · 3% or 150 mM arginine-Cl and 1.5% or 200 mM glycine, and · 3% or 150 mM arginine-Cl and 5.8% or 150 mM magnesium glutamate.

[0161] In one embodiment, the reconstitution solution is water, i.e., sterile water. Thus, the powder or micropellets are reconstituted in water.

[0162] To obtain Composition B, the volume of the aqueous reconstitution buffer that achieves the desired antibody concentration in the antibody in Composition I, i.e., the volume to obtain Composition I having the desired concentration of the antibody such as a concentration of 300 mg / ml, is used.

[0163] It should be understood that the reconstitution of Composition B in aqueous reconstitution requires a specific reconstitution time. In one embodiment, the reconstitution is carried out for a period sufficient to dissolve the antibody in the aqueous solution. Advantageously, it has been shown that the reconstitution time for the spray freeze-dried composition as referred to herein is shorter than the reconstitution time for the spray-dried or freeze-dried Composition B.

[0164] In one embodiment of the provided method, the dry antibody Composition B is obtained by spray drying and is thus a spray-dried powder. Typically, the spray-dried powder is reconstituted in the reconstitution solution for a period of at least 4 hours.

[0165] In one embodiment of the provided method, the dry antibody Composition B is obtained by freeze drying and is thus a freeze-dried powder. Typically, the freeze-dried powder is reconstituted in the reconstitution solution for a period of 1.5 to 4.0 hours.

[0166] In one embodiment of the provided method, the dry antibody Composition B is obtained by spray freeze drying and is thus in the form of spray freeze-dried micropellets. Typically, the spray freeze-dried micropellets are reconstituted in the reconstitution solution in a period of 1 to 2 hours.

[0167] The short reconstitution time enables the preparation of the final Composition I immediately before administering the composition. For example, Composition I can be prepared by a healthcare worker, such as a hospital healthcare worker. The short reconstitution time also enables the preparation of a high-concentration aqueous solution for direct filling into a vial or prefilled syringe. For example, Composition I can be prepared from the reconstituted Composition B, filled into a vial or prefilled syringe at the manufacturing plant, and marketed as an immediate-use liquid product.

[0168] However, the present invention is not limited to the preparation of Composition I immediately before its administration. Since Composition I is stable, it can be prepared well in advance, for example, by the manufacturer of the dry composition. Thereafter, Composition I can be stored for a long period of time. For example, Composition B can be stored for at least 1 month, for example at least 3 months, for example at least 6 months under refrigerated or ambient storage conditions. In one embodiment, the present composition is stored for 1 to 3 months. In an alternative embodiment, Composition I is stored for 3 to 12 months, for example 3 to 6 months. Typically, Composition B is stored refrigerated at a temperature of 2°C to 8°C.

[0169] The definitions and explanations given above in this specification apply hereinafter with the necessary modifications.

[0170] There is further provided a container containing the aqueous antibody Composition I provided herein.

[0171] In one embodiment, the container is a prefilled syringe.

[0172] In an alternative embodiment, the container is a vial.

[0173] In an alternative embodiment, the container is an autoinjector.

[0174] The container can contain any volume of aqueous antibody Composition I considered appropriate. In one embodiment, the container contains about 1 to 4 ml of Composition I, for example 1 to 2 ml of Composition I. For example, the container can contain about 1 ml, 2 ml, 3 ml or 4 ml of Composition I. The volume can depend on the type of container. For example, a vial can contain a volume of up to 50 ml, and a prefilled syringe can contain a volume of up to 2 ml.

[0175] There is also provided a kit comprising the container provided herein and a label or instructions for the administration and use of optionally the aqueous antibody Composition I or the dry antibody Composition B.

[0176] When the container contains the dry antibody composition B, the kit may further contain a reconstitution buffer. Typically, the kit contains a volume of reconstitution buffer sufficient to obtain Composition I.

[0177] Furthermore, provided is the use of the aqueous antibody composition I or the dry antibody composition B provided herein for use in medicine.

[0178] A method for preparing a dry antibody composition B, a) An aqueous composition A, a1) An antibody at a concentration less than 200 mg / ml, such as 10 - 190 mg / ml, such as 20 - 170 mg / ml, such as 50 - 150 mg / ml, such as 80 - 120 mg / ml, etc., based on the total volume of Composition A, as described in relation to the method for preparing Composition I, and a2) At least a surfactant, and a3) At least one cryoprotectant, and a4) At least 0.4% (w / v) arginine or its salt (where the weight (w) is calculated as arginine), such as 0.4 - 3.5% (w / v) arginine, and to provide an aqueous composition A, wherein the ratio of the amount of a1) to the sum of the amounts of a3) and a4) is 11:1 - 11:8, and b) Further provided is a method of spray freeze - drying the composition provided in step a) to give a dry antibody composition B.

[0179] In one embodiment, the dry antibody composition is in the form of micropellets.

[0180] Further provided is a dry antibody composition B obtainable or obtained by the method for preparing a dry antibody composition B.

[0181] Further provided is a dry antibody composition B in the form of micropellets, and the composition B comprises a1) An antibody (as defined herein), and a2) At least one surfactant (as defined herein), and a3) at least one cryoprotectant (as defined herein), such as trehalose, and a4) arginine or a salt thereof (as defined herein) and wherein the ratio of the amount of a1) to the total amount of a3) and a4) is from 11:1 to 11:8, such as from 11:2 to 11:8, and the weight of a4) is calculated as free amino acid.

[0182] In some embodiments, the ratio of the amount of a1) to the total amount of a3) and a4) is from 11:1 to 11:4, such as from 11:2 to 11:4.

[0183] There is further provided a dry antibody composition B in the form of a powder, the composition B comprising a1) an antibody (as defined herein), and a2) at least one surfactant (as defined herein), and a3) at least one cryoprotectant (as defined herein), such as trehalose, and a4) arginine or a salt thereof (as defined herein) and wherein the ratio of the amount of a1) to the total amount of a3) and a4) is from 11:1 to 11:8, such as from 11:2 to 11:8, and the weight of a4) is calculated as free amino acid.

[0184] In some embodiments, the ratio of the amount of a1) to the total amount of a3) and a4) is from 11:1 to 11:4, such as from 11:2 to 11:4.

[0185] There is further provided a container comprising the dry antibody composition B provided herein. For example, the container can be a vial, a prefilled syringe, a cartridge, an ampoule or an autoinjector. In some embodiments, the container is a prefilled syringe or an autoinjector.

[0186] There is also provided the use of the dry antibody composition B provided herein for preparing an aqueous antibody composition I comprising at least 200 mg / ml of an antibody, such as more than 200 mg / ml of an antibody.

[0187] There is further provided an aqueous antibody composition I as described herein. This composition can be used in medicine.

[0188] In one embodiment, the aqueous antibody composition I comprises an antibody at 270 - 350 mg / ml, such as an antibody at 280 - 320 mg / ml.

[0189] In a further embodiment, the aqueous antibody composition I comprises an antibody at at least 250 mg / ml, such as an antibody at at least 300 mg / ml.

[0190] In another embodiment of the provided aqueous antibody composition I, it comprises an antibody at 230 mg / ml - 420 mg / ml, such as an antibody at 260 mg / ml - 400 mg / ml.

[0191] In one embodiment, the aqueous antibody composition I comprises an antibody at about 300 mg / ml.

[0192] Typically, the aqueous antibody composition I has an opalescence of less than 40 Nephelometric Turbidity Units (NTU), such as an opalescence of less than 25 NTU. For example, the opalescence can be less than 15 NTU.

[0193] In one embodiment, the aqueous antibody composition I further comprises at least one surfactant, at least one cryoprotectant, such as trehalose and arginine or its salt.

[0194] In some embodiments, the aqueous antibody composition I is obtained or can be obtained by a method for preparing the aqueous antibody composition I as provided herein.

[0195] Furthermore, there is provided an aqueous composition A as defined in relation to a method for preparing an aqueous antibody composition I comprising an antibody at at least 200 mg / ml, said composition A a1) An antibody of less than 200 mg / ml, such as an antibody of 10 - 190 mg / ml, such as an antibody of 20 - 170 mg / ml, such as an antibody of 50 - 150 mg / ml, such as an antibody of 80 - 120 mg / ml, and a2) At least one surfactant, and a3) At least one cryoprotectant, and a4) Arginine or a salt thereof at least 0.4% (w / v) based on the total volume of the aqueous composition A (where the weight (w) is calculated as arginine), for example, arginine of 0.4 - 3.5% (w / v) and comprising, the ratio of the amount of a1) to the sum of the amounts of a3) and a4) being 11:1 to 11:8.

[0196] The definitions and explanations made for composition A in relation to the method of preparing an aqueous antibody composition apply with the necessary modifications.

[0197] Embodiments The following provides an overview of the embodiments provided. The definitions and explanations given above in this specification apply below with the necessary modifications. 1. A method of preparing an aqueous antibody composition I comprising an antibody of at least 200 mg / ml, comprising a) An aqueous composition A, comprising a1) An antibody at a concentration lower than the concentration of the antibody in the antibody composition I, a2) At least one surfactant, a3) At least one cryoprotectant, a4) Arginine or a salt thereof at least 0.4% (w / v) (where the weight (w) is calculated as arginine), for example, arginine of 0.4 - 3.5% (w / v) comprising, and providing an aqueous composition A in which the ratio of the amount of a1) to the sum of the amounts of a3) and a4) is 11:1 to 11:8; b) Drying the composition provided in step a) to obtain an antibody composition B; and c) Reconstituting the composition dried in step b) to give an aqueous antibody composition I and a method comprising. 2. The method according to embodiment 1, wherein the salt of arginine is arginine-Cl, arginine-aspartate, arginine-glutamate, arginine-sulfate, arginine-acetate, arginine-succinate. 3. The method according to embodiment 2, wherein the salt of arginine is arginine-Cl. 4. The method according to any one of embodiments 1 to 3, wherein at least one cryoprotectant is selected from the group of cryoprotectants consisting of trehalose, sucrose, sorbitol, glycerol and mannitol. 5. The method according to embodiment 4, wherein the cryoprotectant is trehalose. 6. The ratio of the amount of a1) to the sum of the amounts of a3) and a4) is from 11:1 to 11:4, for example from 11:2 to 11:4. The method according to any one of embodiments 1 to 5. 7. The method according to any one of embodiments 1 to 6, wherein the aqueous composition A contains an antibody at less than 200 mg / ml, for example an antibody at 10 to 190 mg / ml, for example an antibody at 20 to 170 mg / ml, for example an antibody at 50 to 150 mg / ml, for example an antibody at 80 to 120 mg / ml. 8. The method according to any one of embodiments 1 to 7, wherein the composition I contains an antibody at more than 200 mg / ml, for example an antibody at 230 mg / ml to 420 mg / ml, for example an antibody at 260 mg / ml to 400 mg / ml. 9. The method according to embodiment 8, wherein the composition I contains an antibody at 270 to 350 mg / ml, for example an antibody at 280 to 320 mg / ml, for example an antibody at 300 mg / ml. 10. The method according to any one of embodiments 1 to 9, wherein the antibody is a monoclonal antibody. 11. The method according to any one of embodiments 1 to 10, wherein at least the surfactant is a polysorbate such as polysorbate 20 or 80 or a poloxamer such as poloxamer 188. 12. The method according to any one of embodiments 1 to 11, wherein the composition A further contains a5) a buffer, for example a histidine, citric acid, acetic acid, phosphoric acid, tris, succinic acid or glycine buffer. 13. The pH value of Composition A is from about 5.0 to 8.0, such as from about 5.5 to 7, the method according to any one of Embodiments 1 to 12. 14. Antibody Composition B is in the form of a powder or micro pellets, the method according to any one of Embodiments 1 to 13. 15. Step b) is carried out by lyophilization or spray drying to give a lyophilized or spray dried powder, the method according to any one of Embodiments 1 to 14. 16. The lyophilized powder is reconstituted in a reconstitution solution over a period of 1.5 to 4.0 hours, or the spray dried powder is reconstituted in a reconstitution solution over a period of at least 4 hours, the method according to Embodiment 15. 17. Step b) is carried out by spray freeze drying to give spray freeze dried micro pellets, the method according to any one of Embodiments 1 to 14. 18. The spray freeze dried micro pellets are reconstituted in a reconstitution solution in a period of 1 to 2 hours, the method according to Embodiment 17. 19. Aqueous Antibody Composition I is a clear and transparent solution, the method according to any one of Embodiments 1 to 18. 20. Aqueous Antibody Composition I obtainable or obtained by the method according to any one of Embodiments 1 to 19. 21. A container containing the aqueous antibody composition I according to Embodiment 20. 22. A prefilled syringe, vial, cartridge, ampoule or autoinjector, the container according to Embodiment 21. 23. A kit comprising the container according to Embodiment 21 or 22 and a label or instructions for administration and use of the aqueous antibody composition I. 24. The aqueous antibody composition I according to Embodiment 21 for use in medicine. 25. A method for preparing a dry antibody composition B, a) An aqueous composition A, a1) An antibody of less than 200 mg / ml, such as an antibody of 10 to 190 mg / ml, such as an antibody of 20 to 170 mg / ml, such as an antibody of 50 to 150 mg / ml, such as an antibody of 80 to 120 mg / ml, and a2) At least a surfactant, a3) at least one cryoprotectant, and a4) at least 0.4% (w / v) arginine or a salt thereof (where the weight (w) is calculated as arginine), for example 0.4 - 3.5% (w / v) arginine, and which contains, and the ratio of the amount of a1) to the total amount of a3) and a4) is 11:1 - 11:8, provides an aqueous composition A, and b) a method of spray freeze-drying the composition provided in step a) to give a dried antibody composition B. 26. The method according to embodiment 25, wherein the dried antibody composition is in the form of micropellets. 27. A dried antibody composition B obtainable or obtainable by the method according to embodiment 25 or 26. 28. A container containing the dried antibody composition B of embodiment 27. 29. Use of the dried antibody composition according to embodiment 26 or 27 for preparing an aqueous antibody composition I containing at least 200 mg / ml of an antibody. 30. An aqueous antibody composition I containing at least 200 mg / ml of an antibody. 31. The aqueous antibody composition I according to embodiment 30, containing 270 - 350 mg / ml of an antibody, for example 280 - 320 mg / ml of an antibody, for example 300 mg / ml of an antibody. 32. The aqueous antibody composition I according to embodiment 30, containing 230 mg / ml - 420 mg / ml of an antibody, for example 260 mg / ml - 400 mg / ml of an antibody. 33. The aqueous antibody composition I according to any one of embodiments 30 - 32, which is a clear and transparent solution. 34. The aqueous antibody composition I according to any one of embodiments 30 - 33, having an opalescence of less than 40 NTU (nephelometric turbidity unit (NTU)), for example less than 25 NTU or less than 15 NTU. 35. The aqueous antibody composition I according to any one of embodiments 30 - 34, containing at least one surfactant, at least one cryoprotectant, and arginine or a salt thereof. 36. The aqueous antibody composition I according to embodiment 35, wherein at least one cryoprotectant is trehalose. 37. The aqueous antibody composition I according to embodiment 35 or 36, wherein at least one surfactant is a polysorbate such as polysorbate 20 or 80 or a poloxamer such as poloxamer 188. 38. The aqueous antibody composition I according to any one of embodiments 30 to 37, which contains histidine. 39. The aqueous antibody composition I according to any one of embodiments 30 to 38, which further contains glycine and / or magnesium glutamate. 40. A dry antibody composition B in the form of micropellets, a1) an antibody (as defined herein), a2) at least one surfactant (as defined herein), a3) at least one cryoprotectant (as defined herein), for example trehalose, a4) arginine or a salt thereof (as defined herein) and the ratio of the amount of a1) to the sum of the amounts of a3) and a4) is 11:1 to 11:8, for example 11:2 to 11:8, and the weight of a4) is calculated as the free amino acid, the dry antibody composition B. 41. A dry antibody composition B in the form of a powder, a1) an antibody (as defined herein), a2) at least one surfactant (as defined herein), a3) at least one cryoprotectant (as defined herein), for example trehalose, a4) arginine or a salt thereof (as defined herein) and the ratio of the amount of a1) to the sum of the amounts of a3) and a4) is 11:1 to 11:8, for example 11:2 to 11:8, and the weight of a4) is calculated as the free amino acid, the dry antibody composition B. 42. Use of the dry antibody composition according to embodiment 40 or 41 for preparing an aqueous antibody composition I containing at least 200 mg / ml of an antibody. 43. An aqueous composition A, a1) An antibody less than 200 mg / ml, such as an antibody of 10 - 190 mg / ml, such as an antibody of 20 - 170 mg / ml, such as an antibody of 50 - 150 mg / ml, such as an antibody of 80 - 120 mg / ml, and a2) at least one surfactant, and a3) at least one cryoprotectant, and a4) at least 0.4% (w / v) of arginine or a salt thereof (wherein the weight (w) is calculated as arginine) based on the total volume of the aqueous composition A, such as 0.4 - 3.5% (w / v) of arginine, and An aqueous composition A, which contains, and the ratio of the amount of a1) to the sum of the amounts of a3) and a4) is 11:1 - 11:8. 44. The salt of arginine is arginine - Cl, arginine - aspartate, arginine - glutamate, arginine - sulfate, arginine - acetate, arginine - succinate, and the aqueous composition A according to embodiment 43. 45. The salt of arginine is arginine - Cl, and the aqueous composition A according to embodiment 44. 46. The at least one cryoprotectant is selected from the group of cryoprotectants consisting of trehalose, sucrose, sorbitol, glycerol, and mannitol, and the aqueous composition A according to any one of embodiments 43 - 45. 47. The cryoprotectant is trehalose, and the aqueous composition A according to embodiment 46. 48. The ratio of the amount of a1) to the sum of the amounts of a3) and a4) is 11:1 - 11:4, such as 11:2 - 11:4, and the aqueous composition A according to any one of embodiments 43 - 47. 49. The aqueous composition A contains an antibody less than 200 mg / ml, such as an antibody of 10 - 190 mg / ml, such as an antibody of 20 - 170 mg / ml, such as an antibody of 50 - 150 mg / ml, such as an antibody of 80 - 120 mg / ml, and the aqueous composition A according to any one of embodiments 43 - 48. 50. The antibody is a monoclonal antibody, and the aqueous composition A according to any one of embodiments 43 - 49. 51. The aqueous composition A according to any one of Embodiments 43 to 50, wherein the surfactant is at least a polysorbate such as polysorbate 20 or 80 or a poloxamer such as poloxamer 188. 52.a5) The aqueous composition A according to any one of Embodiments 43 to 51, further comprising a buffer, for example, a histidine, citric acid, acetic acid, phosphoric acid, Tris, succinic acid or glycine buffer. 53. The aqueous composition A according to any one of Embodiments 43 to 52, wherein the pH value is about 5.0 to 8.0, for example, about 5.5 to 7. 54. Use of the aqueous composition A as defined in any one of Claims 43 to 53 for preparing an aqueous antibody composition I containing at least 200 mg / ml of an antibody.

[0198] All of the reference documents cited throughout this specification are incorporated herein in their entirety with respect to the disclosure specifically mentioned above.

[0199] Hereinafter, the present invention will be further described with reference to the following examples, which are intended to illustrate but not limit the scope of the present invention.

Examples

[0200] Materials and Methods Physical Stability by Size Exclusion Chromatography (SEC) Physical stability was evaluated by periodically measuring the amount of aggregates formed in monoclonal antibody formulations. Samples were measured by eluting about 100 μg of total monoclonal antibody from a TSK-Gel G3000SWXL analytical column (Tosoh Co.) using 40 mM phosphoric acid and 150 mM sodium chloride at pH 7.2 (detection wavelength: 280 nm). Aggregates were defined as high molecular weight species (HMWS) that eluted earlier than non-aggregated mAb monomers; HMWS% was calculated by dividing the amount of aggregates formed by the total of total aggregates + non-aggregated monoclonal antibody molecules.

[0201] SoloVPE for Protein Concentration The total protein concentration was determined by measuring absorbance at 280 nm using a Lunatic instrument (Unchained Labs). Samples for the powder arm were measured without dilution, and samples for the liquid arm were measured after 10 - 15-fold dilution in the formulation buffer. Two measurements were made for each 3 μL sample, and the average value was reported. The molar extinction coefficient, which was antibody-specific, was used for concentration calculation. Results were considered equivalent if the difference was 10% or less.

[0202] Opalescence Samples were analyzed according to the protocol established by Kingsbury JS et al. J Pharm Sci. 2021 Sep;110(9):3176 - 3182. doi:10.1016 / j.xphs.2021.05.005. Epub 2021 May 15. PMID:34004217. Briefly, samples were analyzed by μ - nephelometry using the "detuned" static light scattering channel of a Wyatt DynaPro NanoStar light scattering instrument. Samples were loaded into NanoStar disposable cuvettes with a 10 μl volume. The instrument was set to acquire data with the laser output reduced to 1%. The static scattering detector voltage was recorded and calibrated against a series of polymer bead turbidity standards of 1, 5, 10, 20, 40, and 80 NTU.

[0203] Spray freeze drying Spray freeze drying was performed using process parameters similar to those described by Clenet et al. (Clenet D, Hourquet V, Woinet B, Ponceblanc H, Vangelisti M. A spray freeze dried micropellet based formulation proof - of - concept for a yellow fever vaccine candidate. Eur J Pharm Biopharm. 2019 Sep;142:334 - 343). High - level process parameters used for spray freeze drying:

[0204]

Table 1

[0205] Spray drying High-level process parameters used in spray drying:

[0206]

Table 2

[0207] Example 1: Feasibility of achieving a high-concentration formulation For this feasibility assessment, eight monoclonal antibodies (designated "mAbA", "mAbB", "mAbC", "mAbD", "mAbE", "mAbF", "mAbG" and "mAbH") were used. mAbs A, B and G were of the IgG4 subtype, while the others belonged to the IgG1 subtype. All antibodies were formulated in a starting aqueous composition A consisting of 100 mg / ml antibody, 10 mM histidine, 2% trehalose, 0.05% polysorbate 80 (the ratio of the amount of antibody to the amount of trehalose was 10:2 w / w). The pH of each composition was selected based on the isoelectric point and formulation screening studies. Each composition A was lyophilized to obtain an intermediate dried antibody powder (composition B). This intermediate dried antibody powder was reconstituted using an aqueous solution containing 1.75% or 100 mM arginine, resulting in an aqueous antibody composition (composition I). The reconstitution volume was selected to obtain aqueous antibody compositions I with antibody concentrations of 100, 150, 200, 250, 300, 350 and 400 mg / ml. When measured by SoloVPE, the target protein concentration was achieved for all eight antibodies. For all concentrations and antibodies tested, generally, clear and transparent solutions were obtained (Figure 1). Clear and transparent solutions were obtained for all eight antibodies up to 300 mg / ml. Except for mAb C, clear and transparent solutions were obtained for seven antibodies up to a maximum of 350 mg / ml. For mAb D, mAb G and mAb H, clear and transparent solutions were obtained up to a maximum of 400 mg / ml. An increase in antibody concentration typically involves an increase in HMWS due to molecular crowding phenomena. However, in this example, the increase in antibody concentration from 100 mg / ml to 350 mg / ml was generally accompanied by a slight increase in HMWS (less than 1%) and a slight decrease in solution opalescence (Figure 2). Changes in HMWS below 5% and less than 1% of the HMWS level during concentration increase are both robust indicators of the physical stability of antibody compositions I having an antibody concentration of at least 200 mg / ml. The opalescence of all antibody compositions I having an antibody concentration of at least 200 mg / ml was lower than 20 NTU, which is another robust indicator of optimal solution properties.Antibody composition I containing 200 - 350 mg / ml and including 200 mg / ml and 350 mg / ml provides optimal properties such as a clear and transparent solution, low HMWS, and opalescence.

[0208] Example 2: Optimal composition for aqueous composition A Four monoclonal antibodies (mAb A, B, D, and E) and eleven compositions were evaluated in this example. An overview is provided in Table 1.

[0209] [Table 3]

[0210] All antibodies were formulated in starting aqueous composition A consisting of 100 mg / ml antibody, 10 mM histidine, 0.05% polysorbate 80, 0.5 - 4% trehalose (molecular weight 343), 0.45 - 3.5% arginine (molecular weight 174.2 g / mol). Trehalose was used as a cryoprotectant. This corresponds to a ratio of 11:1 - 11:8% of the amount of antibody to the total amount of the cryoprotectant trehalose and arginine free base, covering different combinations of 0.5 - 4% and 0.45 - 3.5% trehalose and arginine respectively. The pH of these formulations was antibody - specific and was selected during formulation screening to provide optimal stability. The antibody in starting aqueous composition A was lyophilized to obtain an intermediate dried antibody powder (composition B). The results are shown in Table 2.

[0211] [Table 4]

[0212] As shown in Table 2, for all antibodies and compositions, the increase in HMWS in HMWS during lyophilization is only slight (less than 0.7% in all cases), and generally all compositions of antibody / trehalose + arginine from 11:1 to 11:8 w / w are shown to protect the antibody from degradation during lyophilization. An increase in HMWS of 0.7% is generally a robust indicator of the stability of Composition A during lyophilization. From the data, it was also shown that higher levels of trehalose and arginine, such as compositions of 11:2 to 11:8, are generally favorable for antibody stability. The inventors also observed that the starting aqueous Composition A (black bars in Figure 3) containing at least 0.45% arginine (MW 174.2 g / mol) generally had a smaller increase in HMWS during lyophilization compared to the composition of trehalose only (gray bars in Figure 3). Evaluation of the short-term storage stability of the intermediate lyophilized antibody (Composition B) at 25 °C showed that HMWS increased slightly (generally <1% for mAbA, D, E and generally <2% for mAbB) at the time of storage at 25 °C for 1 month compared to the initial time point, suggesting that generally all compositions provide protection during storage of the intermediate lyophilized antibody (Table 3).

[0213]

Table 5

[0214] It was also shown that higher levels of trehalose and arginine, such as the composition of 11:2 to 11:8, were generally favorable for antibody stability during storage at 25 °C from the data. From the evaluation of the accelerated stability of the intermediate lyophilized antibody (Composition B) at 40 °C, the compositions containing at least 0.45% arginine (MW 174.2 g / mol) (black bars in Figure 4) generally showed a smaller increase in HMWS compared to the composition of trehalose only (gray bars in Figure 4). Subsequently, the intermediate dried antibody powder (Composition B) was reconstituted with water for injection (WFI). The reconstitution volume was selected to obtain an aqueous antibody composition I with an antibody concentration of 300 mg / ml (Composition I). For all antibodies and compositions shown in Figure 5, clear and transparent aqueous solutions with a target protein concentration of 300 mg / ml were obtained. When the HMWS in the 300 mg / ml antibody composition I was compared with the HMWS in the 100 mg / ml starting aqueous antibody composition A (both having the same weight ratio of antibody to trehalose and arginine), generally a slight increase in HMWS was shown as a result of a three-fold increase in antibody concentration. The change in HMWS between antibody composition I and antibody composition A was less than 0.5% for mAbA, D, and E. mAb B showed a high increase in HMWS (less than 4%), but this was generally alleviated at 11:2 to 11:8 w / w (Figure 6), and the change in HMWS was generally less than 2%. The evaluation of the accelerated stability of the 300 mg / ml antibody composition I showed a slight increase in HMWS (<2%) at 40 °C for 2 weeks compared to the initial time point, suggesting that generally all compositions provide protection for the antibody in Composition I (Figure 10). 40 °C corresponds to the standard conditions used for the accelerated testing of biological formulations and is not the intended storage temperature for Composition I. The <2% increase in HMWS at 40 °C for 2 weeks is a small change and is generally considered to indicate high physical stability.

[0215] Example 3: Optimal Reconstitution Solution for Intermediate Dried Antibody Powder Six monoclonal antibodies (designated mAb A, B, D, E, G, and H) were used for this evaluation. All antibodies were formulated in starting aqueous composition A consisting of 100 mg / ml antibody, 10 mM histidine, 2% trehalose, 0.05% polysorbate 80 (formulations F12 - F15), corresponding to approximately 11:2 w / w, or consisting of 100 mg / ml antibody, 10 mM histidine, 3% trehalose, 0.05% polysorbate 80 (formulations 16 - 18), corresponding to approximately 11:3 w / w. The antibodies in starting aqueous composition A were lyophilized to obtain an intermediate dried antibody powder (composition B). Subsequently, the dried antibody powder was reconstituted using 1.5% or 200 mM glycine (F12), 3% or 150 mM arginine-Cl (F13), 6.4% or 200 mM arginine-glutamate (F14), 5.8% or 150 mM magnesium glutamate (F15), 3% or 150 mM arginine-Cl (F16), 3% or 150 mM arginine-Cl and 1.5% or 200 mM glycine (F17) or 3% or 150 mM arginine-Cl and 5.8% or 150 mM magnesium glutamate (F17). The reconstitution volume was selected to obtain an aqueous antibody composition I with an antibody concentration of 300 mg / ml. For all antibodies and compositions, clear and transparent aqueous solutions with a target antibody concentration of 300 mg / ml were obtained (Figure 8). Evaluation of the accelerated stability of the 300 mg / ml antibody composition I showed a slight increase in HMWS (<2%) after 2 weeks at 40 °C compared to the initial time point, suggesting that generally all compositions protect the antibody in composition I (Figure 9). 40 °C corresponds to standard conditions used for accelerated testing of biological formulations and is not the intended storage temperature for composition I. An increase in HMWS of <2% at 40 °C over 2 weeks is a small change and is generally considered to indicate high physical stability. These changes in HMWS are similar to the observations in Figure 4 containing trehalose and arginine-Cl. This may generally suggest that reconstitution solutions containing glycine, arginine-Cl, guanidinium-Cl, arginine-glutamate, and magnesium glutamate are effective in providing protection to the antibody in composition I.

[0216] Example 4: Evaluation of spray drying and spray freeze drying for producing intermediate dried antibody powder For this evaluation, two monoclonal antibodies (designated mAbB and I) and two compositions for each antibody (F20 and F21 for mAb I, and F22 and F23 for mAb B) were used. The starting aqueous composition A of mAb B consisted of (F22) 100 mg / ml of antibody, 0.06% polysorbate 80, 1.75% arginine (equivalent to approximately 11:2 w / w of antibody:trehalose + arginine) and (F23) 100 mg / ml of antibody, 0.06% polysorbate 80, 2% trehalose, 0.9% arginine (equivalent to approximately 11:3 w / w of antibody:trehalose + arginine). The starting aqueous composition A of mAb I consisted of (F20) 100 mg / ml of antibody, 10 mM histidine, 0.06% polysorbate 80, 3% trehalose, 0.9% arginine (equivalent to approximately 11:4 w / w) and (F21) 100 mg / ml of antibody, 10 mM histidine, 0.06% polysorbate 80, 2% trehalose, 0.45% arginine (equivalent to approximately 11:2.5 w / w). Both antibodies in each starting composition A were spray-dried and spray-freeze-dried to obtain intermediate dry antibody powders (composition B). As shown in Figure 10, spray-drying resulted in a loose dry powder and spray-freeze-drying resulted in micro-pellets. The intermediate dry antibody powders (composition B) from both drying techniques were reconstituted with water for injection. The reconstitution volume was selected to obtain an aqueous antibody composition I with an antibody concentration of 300 mg / ml. After reconstitution of the spray-freeze-dried micro-pellets, a clear and transparent aqueous solution with a target antibody concentration of 300 mg / ml was obtained for both the antibody (mAb and I) and the compositions (F22 and F23 for mAb and F20 and F21 for mAb I) (Figure 11). The opalescence of the resulting solutions was <5 NTU for both compositions of mAb B, indicating generally acceptable solution properties. The reconstitution time for reconstituting the micro-pellets was generally 1 - 2 hours. After reconstitution of the spray-dried powder, an antibody concentration of 300 mg / ml was achieved, while the reconstitution time generally exceeded 4 hours and the resulting solutions were generally highly turbid (>20 NTU) and generally lacked clarity and transparency.From the evaluation of the short-term and long-term storage stabilities of the intermediate spray-dried powder and the spray-freeze-dried micropellets (Composition B), generally, after short-term storage at 25°C or long-term storage at 5°C, a significant increase in HMWS was not shown (Figure 12). From the evaluation of the short-term storage and accelerated stability of Composition I at 300 mg / ml obtained as a result of the reconstitution of the spray-dried powder and the spray-freeze-dried micropellets, generally, after short-term storage at 5°C or 25°C for 3 months or at 40°C for 1 month, a significant increase in HMWS was not shown (Figure 13).

Claims

1. A method for preparing an aqueous antibody composition I containing at least 200 mg / ml of antibody, wherein a) an aqueous composition A, a1) An antibody at a concentration lower than the concentration of the antibody in the antibody composition I, a2) At least one surfactant, a3) At least one type of antifreeze, a4) At least 0.4% (w / v) of arginine or its salt (where weight (w) is calculated as arginine), for example 0.4–3.5% (w / v) of arginine A step of providing an aqueous composition A, which includes a1) and the ratio of the amount of a3) and the sum of the amounts of a4) is 11:1 to 11:

8. b) A step of drying the composition provided in step a) to obtain antibody composition B, c) A step of reconstituting the composition dried in step b) to give the aqueous antibody composition I. A method that includes this.

2. The method according to claim 1, wherein the salt of arginine is arginine-Cl, arginine-aspartate, arginine-glutamate, arginine-sulfate, arginine-acetate, or arginine-succinate.

3. The method according to claim 1, wherein the at least one antifreeze agent is selected from the group consisting of trehalose, sucrose, sorbitol, glycerol, and mannitol.

4. The method according to claim 1, wherein the ratio of the amount of a1) to the sum of the amounts of a3) and a4) is 11:1 to 11:4, for example, 11:2 to 11:

4.

5. The method according to claim 1, wherein the aqueous composition A contains an antibody in an amount of less than 200 mg / ml, for example, an antibody in an amount of 10 to 190 mg / ml, for example, an antibody in an amount of 20 to 170 mg / ml, for example, an antibody in an amount of 50 to 150 mg / ml, for example, an antibody in an amount of 80 to 120 mg / ml.

6. Composition I contains an antibody in a concentration of 230 mg / ml to 420 mg / ml, for example, 260 mg / ml to Contains 400 mg / ml of antibody, or The method according to claim 1, wherein composition I comprises an antibody in a concentration of 270 to 350 mg / ml, for example, an antibody in a concentration of 280 to 320 mg / ml, for example, an antibody in a concentration of 300 mg / ml.

7. a) The antibody is a monoclonal antibody, b) The surfactant is at least a polysorbate such as polysorbate 20 or 80, or a poloxamer such as poloxamer 188. c) The pH of composition A is about 5.0 to 8.0, for example, about 5.5 to 7, and / or d) Composition A further comprises a5) a buffering agent, for example, histidine, citric acid, acetic acid, phosphoric acid, tris, succinic acid, or glycine buffering agent, according to claim 1.

8. The method according to claim 1, wherein the antibody composition B is in the form of a powder or micropellets.

9. The method according to claim 1, wherein step b) is carried out by freeze-drying or spray-drying to give a freeze-dried or spray-dried powder, and optionally the freeze-dried powder is reconstituted in a reconstitution solution for a period of 1.5 to 4 hours, or the spray-dried powder is reconstituted in a reconstitution solution for a period of at least 4 hours.

10. The method according to claim 1, wherein step b) is carried out by spray freeze-drying to give spray freeze-dried micropellets.

11. The method according to claim 10, wherein in step c), the spray freeze-dried micropellets are reconstituted in a reconstitution solution over a period of 1 to 2 hours.

12. Aqueous antibody composition I obtained or obtainable by the method described in any one of claims 1 to 11.

13. For example, a container containing the aqueous antibody composition I according to claim 12, which is a pre-filled syringe, vial, cartridge, ampoule, or automatic injection device.

14. A method for preparing dried antibody composition B, a) Aqueous composition A, a1) Antibodies less than 200 mg / ml, for example, antibodies between 10 and 190 mg / ml, for example, antibodies between 20 and 170 mg / ml, for example, antibodies between 50 and 150 mg / ml, for example, antibodies between 80 and 120 mg / ml, a2) At least one surfactant, a3) At least one type of antifreeze agent, a4) At least 0.4% (w / v) arginine or its salt (where weight (w) is calculated as arginine), for example 0.4–3.5% (w / v) arginine and The present invention provides an aqueous composition A which includes, and the ratio of the amount of a1) to the sum of the amounts of a3) and a4) is 11:1 to 11:8, and b) A method comprising spray freeze-drying the composition provided in step a) to obtain the dried antibody composition B.

15. The method according to claim 14, wherein the dried antibody composition is in the form of micropellets.

16. The arginine salt is arginine-Cl, arginine-aspartate, arginine-glutamate, arginine-sulfate, arginine-acetate, or arginine-succinate, for example, the arginine salt is arginine-Cl, as described in claim 14. The method.

17. The method according to claim 14, wherein the at least one antifreeze agent is selected from the group of antifreeze agents consisting of trehalose, sucrose, sorbitol, glycerol, and mannitol, and for example, the antifreeze agent is trehalose.

18. The method according to claim 14, wherein the ratio of the amount of a1) to the sum of the amounts of a3) and a4) is 11:1 to 11:4, for example, 11:2 to 11:

4.

19. The method according to claim 14, wherein the aqueous composition A comprises 50 to 150 mg / ml of antibody, for example, 80 to 120 mg / ml of antibody.

20. The method according to claim 14, wherein composition A further comprises a5) a buffering agent, such as histidine, citric acid, acetic acid, phosphoric acid, tris, succinic acid, or glycine buffering agent.

21. The method according to claim 14, wherein the antibody is a monoclonal antibody.

22. A dried antibody composition B obtained or obtainable by the method described in any one of claims 14 to 21.

23. The use of the dried antibody composition B according to claim 14 or the aqueous antibody composition A according to claim 1 for preparing an aqueous antibody composition I containing at least 200 mg / ml of antibody.

24. The aqueous composition A according to claim 1.

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