Pharmaceutical preparations containing anti-OX40L antibodies

Abstract

This disclosure provides aqueous pharmaceutical formulations comprising an anti-OX40L antibody or its antigen-binding fragment for treating or preventing OX40L-mediated diseases or conditions in humans. This disclosure also provides containers and kits comprising the aqueous pharmaceutical formulations, as well as the use of these aqueous pharmaceutical formulations in the treatment or prevention of OX40L-mediated diseases or conditions in humans.

Description

[Background technology]

[0001] OX40 ligand (OX40L) is a TNF family member and a 34 kDa type II transmembrane protein. The crystallized complex of human OX40 and OX40L consists of one OX40L (trimer) and three OX40 monomers. The human extracellular domain is 42% homologous to mouse OX40L.

[0002] OX40L is not constitutively expressed, but can be induced on professional APCs such as B cells, dendritic cells (DCs), and macrophages. Other cell types, such as Langerhans cells, endothelial cells, smooth muscle cells, mast cells, and natural killer (NK) cells, can also be induced to express OX40L. T cells can also express OX40L. The OX40L receptor, OX40, is activated on T cells (CD4). + and CD8 + T cells (Th2, Th1, and Th17 cells), as well as CD4 + Foxp3 + It is expressed on cells even in the absence of activation.

[0003] The interaction between OX40 and OX40L occurs during T cell-DC interaction two or three days after antigen recognition. After leaving the DC, OX40-expressing T cells can interact with OX40L-expressing cells other than DCs, and may receive OX40 signals from these cells, which can provide essential signals for memory T cell generation, enhancement of the Th2 response, and prolongation of the inflammatory response. OX40 signaling into responder T cells makes them resistant to Treg-mediated suppression.

[0004] Anti-OX40L monoclonal antibodies (anti-OX40L mAbs) may be clinically useful for the treatment or prevention of OX40L-mediated diseases or conditions. Exemplary anti-human OX40L (hOX40L) antibodies and fragments for the treatment or prevention of hOX40L-mediated diseases or conditions in humans, as well as their medical applications, are described, among others, in WO2015 / 132580 and U.S. Patent No. 9139653.

[0005] Therefore, there is a demand for anti-OX40L antibody pharmaceuticals that are suitable for administration to subjects requiring anti-OX40L antibody preparations, while simultaneously maintaining an appropriate protein structure throughout their shelf life and ensuring appropriate and accurate drug delivery to achieve the desired efficacy for the treatment or prevention of OX40L-mediated diseases or conditions at the time of administration.

[0006] Aqueous (liquid)-based formulations are typically exemplary drug formulations for therapeutic antibodies, such as monoclonal antibodies (mAbs) and mAb-based therapeutics, which can be used for intravenous (IV) or subcutaneous (SC) delivery of mAbs or mAb-based therapeutics to achieve maximum bioavailability. However, therapeutic antibodies in aqueous solutions are susceptible to degradation, aggregation, or undesirable chemical modifications unless the solution is properly formulated. The stability of antibodies in aqueous formulations depends not only on the types of excipients used in the formulation, but also on the amounts and ratios of the excipients to each other. Apart from stability, other factors such as the suitable viscosity and turbidity of the formulation, the concentration of antibody that can be contained by a particular formulation, the visual quality or appearance of the formulation, and other properties that allow the formulation to be conveniently administered to the target are factors that must be considered when formulating therapeutic antibodies. Thus, developing a well-formulated mAb, such as a well-formulated anti-OX40L mAb, is challenging.

[0007] This disclosure addresses the need for stable aqueous pharmaceutical formulations and pharmaceuticals containing high or low concentrations of anti-OX40L antibody. [Overview of the project]

[0008] This disclosure is based on the discovery of specific aqueous pharmaceutical formulations that function particularly well to stabilize anti-human OX40L (hOX40L) antibodies at a wide range of concentrations, including high concentrations of anti-human OX40L (hOX40L) antibodies. The formulations described herein provide advantageous turbidity reduction and increased stabilization of hOX40L antibodies when exposed to metal and / or stir-induced stress. The aqueous pharmaceutical formulations disclosed herein are stable when packaged in containers such as vials or injection devices.

[0009] This disclosure provides an aqueous pharmaceutical formulation comprising an anti-hOX40 ligand (OX40L) antagonist antibody or its antigen-binding fragment, which is suitable for delivery to a subject requiring it via an injectable route. This disclosure also relates to pharmaceutical unit doses, containers, and kits, including the aqueous pharmaceutical formulation, and to the use of the aqueous formulation for treating hOX40-mediated diseases or conditions in a subject requiring it.

[0010] This disclosure provides an aqueous pharmaceutical formulation comprising an anti-hOX40L antibody or its antigen-binding fragment, which is suitable for delivery to a subject requiring it via an injectable route. This disclosure also relates to pharmaceutical unit doses, containers, and kits, including the aqueous pharmaceutical formulation, and to the use of the aqueous formulation for treating hOX40L-mediated diseases or conditions in a subject requiring it.

[0011] In one embodiment, the embodiments of the present disclosure relate to an aqueous pharmaceutical formulation comprising (a) an antibody or antigen-binding fragment that specifically binds to an OX40 ligand (OX40L), (b) a stabilizer, (c) a surfactant, and (d) a buffer containing histidine or a histidine salt, wherein the pH of the aqueous pharmaceutical formulation is in the range of 5.5±0.2 to about 6.5±0.2, and the antibody or antigen-binding fragment comprises the heavy chain complementarity-determining region (HCDR) 1 of SEQ ID NO: 42, the HCDR 2 of SEQ ID NO: 44, and the HCDR 3 of SEQ ID NO: 46, the light chain complementarity-determining region (LCDR) 1 of SEQ ID NO: 56, the LCDR 2 of SEQ ID NO: 58, and the LCDR 3 of SEQ ID NO: 60, and the aqueous pharmaceutical formulation is suitable for parenteral administration to mammalian subjects.

[0012] In one embodiment, the embodiments of the present disclosure relate to an aqueous pharmaceutical formulation comprising (a) an anti-OX40 ligand (OX40L) antagonist antibody or antigen-binding fragment, (b) a stabilizer, (c) a surfactant, and (d) a buffer containing histidine or a histidine salt, wherein the pH of the aqueous pharmaceutical formulation is in the range of 5.5±0.2 to about 6.5±0.2, and the antibody or antigen-binding fragment comprises the heavy chain complementarity-determining region (HCDR) 1 of SEQ ID NO: 42, the HCDR 2 of SEQ ID NO: 44, and the HCDR 3 of SEQ ID NO: 46, the light chain complementarity-determining region (LCDR) 1 of SEQ ID NO: 56, the LCDR 2 of SEQ ID NO: 58, and the LCDR 3 of SEQ ID NO: 60, and the aqueous pharmaceutical formulation is suitable for parenteral administration to mammalian subjects.

[0013] In a particular embodiment, the antibody or its antigen-binding fragment is amriterimab or a variant thereof.

[0014] In one embodiment, embodiments of the present disclosure relate to an aqueous pharmaceutical formulation comprising (a) an anti-OX40 ligand (OX40L) antagonist antibody or antigen-binding fragment, (b) a stabilizer, (c) a chelating agent, (d) a surfactant, and (e) a buffer containing histidine or a histidine salt, wherein the pH of the aqueous pharmaceutical formulation is in the range of 5.5 ± 0.2 to about 6.5 ± 0.2, and the antibody or antigen-binding fragment comprises the heavy chain complementarity-determining region (HCDR) 1 of SEQ ID NO: 42, the HCDR 2 of SEQ ID NO: 44, and the HCDR 3 of SEQ ID NO: 46, the light chain complementarity-determining region (LCDR) 1 of SEQ ID NO: 56, the LCDR 2 of SEQ ID NO: 58, and the LCDR 3 of SEQ ID NO: 60, and the aqueous pharmaceutical formulation is suitable for parenteral administration to mammalian subjects. In certain embodiments, the antibody or antigen-binding fragment is amriterimab or a variant thereof.

[0015] In another embodiment, embodiments of the present disclosure relate to an aqueous pharmaceutical formulation comprising (a) an antibody or antigen-binding fragment that specifically binds to an OX40 ligand (OX40L), (b) a stabilizer, (c) a chelating agent, (d) a surfactant, and (e) a buffer comprising histidine or a histidine salt, wherein the pH of the aqueous pharmaceutical formulation is in the range of 5.5 ± 0.2 to about 6.5 ± 0.2, and the antibody or antigen-binding fragment comprises the heavy chain complementarity-determining region (HCDR) 1 of SEQ ID NO: 42, the HCDR 2 of SEQ ID NO: 44, and the HCDR 3 of SEQ ID NO: 46, the light chain complementarity-determining region (LCDR) 1 of SEQ ID NO: 56, the LCDR 2 of SEQ ID NO: 58, and the LCDR 3 of SEQ ID NO: 60, and the aqueous pharmaceutical formulation is suitable for parenteral administration to mammalian subjects. In certain embodiments, the antibody or antigen-binding fragment is amriterimab or a variant thereof.

[0016] In certain embodiments, the antibody or its antigen-binding fragment contained in the aqueous pharmaceutical formulation is amriterimab or a variant thereof. In certain embodiments, the antibody or its antigen-binding fragment is present at concentrations ranging from about 28 mg / mL to about 138 mg / mL, or from 31 mg / mL to 125 mg / mL. In one embodiment, the antibody or its antigen-binding fragment is present at concentrations of 28 mg / mL, 31 mg / mL, 62.5 mg / mL, 125 mg / mL, or 138 mg / mL. In one embodiment, the antibody or its antigen-binding fragment is present at concentrations of 28 mg / mL, 31 mg / mL, 125 mg / mL, or 138 mg / mL. In certain embodiments, the antibody or its antigen-binding fragment is present at concentrations ranging from 75 mg / mL to 125 mg / mL. In some embodiments, the antibody or its antigen-binding fragment is present in the aqueous pharmaceutical formulation at concentrations of 125 mg / mL to 138 mg / mL. In one embodiment, the antibody or its antigen-binding fragment is present at a concentration of 125 mg / mL.

[0017] In certain embodiments, the antibody or its antigen-binding fragment is administered to the subject as an initial dose of 500 mg, followed by one or more secondary doses of 250 mg at each q4w. In certain embodiments, the loading dose includes a 2 × 2 mL injection of 250 mg. In certain embodiments, one or more secondary doses each include a 1 × 2 mL injection of 250 mg.

[0018] In a particular embodiment, the antibody or its antigen-binding fragment is administered to the subject as an initial dose of 500 mg, followed by one or more secondary doses of 250 mg every 12 weeks (q12w). In a particular embodiment, the antibody or its antigen-binding fragment is administered to the subject as an initial dose of 250 mg, followed by one or more secondary doses of 125 mg every 12 weeks (q12w).

[0019] In certain embodiments, the antibody or its antigen-binding fragment is administered to the subject as an initial dose of 250 mg, followed by one or more secondary doses of 250 mg at each q4w. In certain embodiments, the loading dose comprises a 1 × 2 mL injection of 250 mg of the antibody or its antigen-binding fragment and a 1 × 2 mL placebo. In certain embodiments, one or more secondary doses each comprise a 1 × 2 mL injection of 250 mg.

[0020] In certain embodiments, the antibody or its antigen-binding fragment is administered to the subject as an initial dose of 15 mg, followed by one or more secondary doses of 125 mg at each q4w. In certain embodiments, the loading dose comprises a 1 × 2 mL injection of 125 mg of the antibody or its antigen-binding fragment and a 1 × 2 mL placebo. In certain embodiments, one or more secondary doses each comprise a 1 × 2 mL injection of 125 mg.

[0021] In a particular embodiment, the stabilizer in the pharmaceutical formulation is mannitol and / or sucrose. In a particular embodiment, the stabilizer in the pharmaceutical formulation is sucrose. In one embodiment, at least one stabilizer is sucrose present in an amount of 220 mM ± 33 mM. In one embodiment, at least one stabilizer is sucrose present in an amount of 220 mM.

[0022] In certain embodiments, the surfactant in the pharmaceutical formulation is selected from polysorbate 80. In one embodiment, the aqueous pharmaceutical formulation contains 0.01% (w / v) to 0.1% (w / v) of polysorbate 80. In one embodiment, the aqueous pharmaceutical formulation contains 0.02% (w / v) to 0.1% (w / v) of polysorbate 80. In one embodiment, the aqueous pharmaceutical formulation contains 0.02% (w / v) to 0.06% (w / v) of polysorbate 80. In another embodiment, the aqueous pharmaceutical formulation contains 0.04% (w / v) of polysorbate 80. In yet another embodiment, the aqueous pharmaceutical formulation contains 0.06% (w / v) of polysorbate 80.

[0023] In certain embodiments, the buffer solution contained in the pharmaceutical formulation contains 10 mM ± 1.5 mM of L-histidine or histidine hydrochloride. In certain embodiments, the buffer solution contained in the pharmaceutical formulation contains 20 mM of L-histidine or histidine hydrochloride.

[0024] In certain embodiments, the pharmaceutical formulation further comprises a chelating agent selected from the group consisting of ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), and their salts, and any combination thereof. In certain embodiments, the chelating agent contains 10 μM of EDTA or 10 μM of DTPA.

[0025] In one embodiment, the aqueous pharmaceutical formulation contains 28 ± 4.2 mg / mL to 138 mg / mL ± 20.7 mg / mL of an antibody or an antigen-binding fragment thereof, 10 mM ± 1.5 mM of L-histidine or histidine hydrochloride, 220 mM of sucrose, 0.06% (w / v) of polysorbate 80, optionally 10 μM of EDTA or 10 μM of DTPA, water, and the pH of the aqueous pharmaceutical formulation is from about 5.8 to about 6.2.

[0026] In one embodiment, the aqueous pharmaceutical formulation contains 125 mg / mL to 138 mg / mL of an antibody or an antigen-binding fragment thereof, 20 mM of L-histidine or histidine hydrochloride, 220 mM of sucrose, 0.04% (w / v) of polysorbate 80, and water, and the pH of the aqueous pharmaceutical formulation is from about 5.8 to about 6.2.

[0027] In some embodiments, the aqueous pharmaceutical formulation does not contain or is essentially free of particles. In some embodiments, the aqueous pharmaceutical formulation does not contain or is substantially free of sodium chloride. In some embodiments, the aqueous pharmaceutical formulation does not contain or is substantially free of arginine.

[0028] In some embodiments, less than 5% of the antibody was detected in aggregate form after storage at 40°C for 28 days and detected by size exclusion high-performance liquid chromatography.

[0029] In some embodiments, at least 91% of the antibodies in the aqueous pharmaceutical formulation, as measured by size exclusion chromatography, have a natural conformation after 28 days at 40°C. In some embodiments, at least 94% of the antibodies in the aqueous pharmaceutical formulation, as measured by size exclusion chromatography, have a natural conformation after 8 weeks at 25°C. In some embodiments, at least 98% of the antibodies, as measured by size exclusion chromatography, have a natural conformation after 16 weeks at 5°C or -65°C.

[0030] In some embodiments, at least 55% of the antibody in the aqueous pharmaceutical formulation is the major charge variant of the antibody after 28 days at 40°C, as measured by imaging capillary isoelectric focusing (iCIEF). In some embodiments, at least 65% of the antibody is the major charge variant of the antibody after 16 weeks at 25°C, as measured by imaging capillary isoelectric focusing (iCIEF). In some embodiments, at least 70% of the antibody is the major charge variant of the antibody after 16 weeks at 5°C, as measured by imaging capillary isoelectric focusing (iCIEF). In some embodiments, at least 70% of the antibody is the major charge variant of the antibody after 16 weeks at -65°C, as measured by imaging capillary isoelectric focusing (iCIEF).

[0031] In one particular embodiment, the aqueous pharmaceutical formulation is stable when stored at 5°C or 25°C for at least 3 months. In one embodiment, the aqueous pharmaceutical formulation is stable when stored at 40°C for at least 2 months. In one embodiment, the aqueous pharmaceutical formulation is stable when stored at 5°C for at least 1 year.

[0032] In certain embodiments, the aqueous pharmaceutical formulation is stable when stored at -65°C, 5°C, or 25°C for at least 16 weeks. In one embodiment, the aqueous pharmaceutical formulation is stable when stored at 40°C for at least 8 weeks. In one embodiment, the aqueous pharmaceutical formulation is stable when stored at 5°C for at least 1 year. In certain embodiments, the aqueous pharmaceutical formulation is stable during freezing and thawing.

[0033] In certain embodiments, the aqueous pharmaceutical formulation according to this disclosure is contained in a container. In certain embodiments, the aqueous pharmaceutical formulation is suitable for subcutaneous delivery.

[0034] In one embodiment, this application relates to a pharmaceutical unit dosage form suitable for parenteral administration to mammalian subjects, comprising an aqueous pharmaceutical formulation disclosed herein in a suitable container. In a particular embodiment, the aqueous pharmaceutical formulation is suitable for intravenous, subcutaneous, or intramuscular administration. In one embodiment, the suitable container is a pre-filled syringe. In one embodiment, the suitable container is a pre-filled pen or auto-injector.

[0035] In another embodiment, embodiments of the present disclosure relate to a sealed container containing an aqueous pharmaceutical formulation disclosed herein. In some embodiments, the sealed container is a vial, syringe, microinfuser, pen delivery device, or auto-injector. In another embodiment, the sealed container is a single or multi-chamber syringe. In yet another embodiment, the sealed container is a pre-filled syringe containing 2.25 ml of the aqueous pharmaceutical formulation.

[0036] In yet another embodiment, embodiments of the present disclosure relate to a pre-filled syringe containing an aqueous pharmaceutical formulation comprising an antibody or its antigen-binding fragment that specifically binds to an OX40 ligand (OX40L) in a concentration of approximately 28 mg / mL to approximately 138 mg / mL, 10 mM L-histidine or histidine hydrochloride, 220 mM sucrose, 0.06% (w / v) polysorbate 80, and water, wherein the pH of the aqueous pharmaceutical formulation is 6.0. In some embodiments, the aqueous pharmaceutical formulation contained in the pre-filled syringe may further contain 10 μM EDTA or 10 μM DTPA. In some embodiments, the pre-filled syringe is equipped with a safety system such as a needle guard, for example, a BD UltraSafe® or BD UltraSafe Plus® needle guard. In some embodiments, the aqueous pharmaceutical formulation contained in the pre-filled syringe contains an antibody or its antigen-binding fragment that specifically binds to an OX40 ligand (OX40L) in a concentration of 31 mg / mL to 125 mg / mL. In a particular embodiment, the aqueous pharmaceutical formulation contained in a pre-filled syringe contains amliterimab or a variant thereof in an amount of approximately 31 mg / mL to approximately 125 mg / mL. In a particular embodiment, the aqueous pharmaceutical formulation contained in a pre-filled syringe contains amliterimab or a variant thereof in an amount of 125 mg / mL ± 18.75 mg / mL, or approximately 125 mg / mL. In one embodiment, the pre-filled syringe contains approximately 125 mg of amliterimab in a 1 mL solution (125 mg / mL ± 18.75 mg / mL). In one embodiment, the pre-filled syringe contains approximately 250 mg of amliterimab in a 2 mL solution (125 mg / mL ± 18.75 mg / mL).

[0037] In yet another embodiment, embodiments of the present disclosure relate to a pre-filled pen or auto-injector containing an aqueous pharmaceutical formulation comprising an antibody or antigen-binding fragment thereof that specifically binds to an OX40 ligand (OX40L) in a concentration of about 28 mg / mL to about 138 mg / mL, 10 mM L-histidine or histidine hydrochloride, 220 mM sucrose, 0.06% (w / v) polysorbate 80, and water, wherein the pH of the aqueous pharmaceutical formulation is 6.0. In some embodiments, the aqueous pharmaceutical formulation contained in the pre-filled pen or auto-injector may further contain 10 μM EDTA or 10 μM DTPA. In some embodiments, the aqueous pharmaceutical formulation contained in the pre-filled pen or auto-injector contains an antibody or antigen-binding fragment thereof that specifically binds to an OX40 ligand (OX40L) in a concentration of 31 mg / mL to 125 mg / mL. In certain embodiments, the aqueous pharmaceutical formulation contained in a pre-filled pen or auto-injector contains approximately 31 mg / mL to approximately 125 mg / mL of amliterimab or a variant thereof. In certain embodiments, the aqueous pharmaceutical formulation contained in a pre-filled pen or auto-injector contains 125 mg / mL ± 18.75 mg / mL of amliterimab or a variant thereof. In one embodiment, the pre-filled pen or auto-injector contains approximately 125 mg of amliterimab in 1 mL of solution (125 mg / mL ± 18.75 mg / mL). In one embodiment, the pre-filled pen or auto-injector contains approximately 250 mg of amliterimab in 2 mL of solution (125 mg / mL ± 18.75 mg / mL).

[0038] In yet another embodiment, the embodiments of the present disclosure relate to a pre-filled syringe containing an aqueous pharmaceutical formulation comprising 125 mg / mL to 138 mg / mL of amriterimab or a variant thereof, 20 mM L-histidine or histidine hydrochloride, 220 mM sucrose, 0.04% (w / v) polysorbate 80, and water, wherein the pH of the aqueous pharmaceutical formulation is 6.0.

[0039] In one embodiment, the pre-filled pen or auto-injector contains a single-dose formulation. In one embodiment, the pre-filled pen or auto-injector is sleeve-operated. In one embodiment, the pre-filled pen or auto-injector comprises a fixedly positioned syringe. In one embodiment, the pre-filled pen or auto-injector provides audible feedback at the end of injection and / or after injection. In one embodiment, the pre-filled pen or auto-injector comprises an anti-rotation mechanism.

[0040] In yet another embodiment, embodiments of the present disclosure relate to a kit comprising a sealed container containing an aqueous pharmaceutical formulation disclosed herein. In one embodiment, the sealed container is a pre-filled syringe containing the aqueous pharmaceutical formulation disclosed herein. In one embodiment, the sealed container is a pre-filled pen or auto-injector containing the aqueous pharmaceutical formulation disclosed herein. In one embodiment, the pre-filled pen or auto-injector contains a single-dose formulation. In one embodiment, the pre-filled pen or auto-injector is sleeve-operated. In one embodiment, the pre-filled pen or auto-injector comprises a fixedly positioned syringe. In one embodiment, the pre-filled pen or auto-injector provides audible feedback at the end of injection and / or after injection. In one embodiment, the pre-filled pen or auto-injector comprises an anti-rotation mechanism.

[0041] In yet another embodiment, embodiments of the present disclosure relate to a kit comprising a sealed container containing any aqueous pharmaceutical formulation disclosed herein, and at least one separate injection device for delivering the aqueous pharmaceutical formulation to a mammalian subject requiring it. In one embodiment, the injection device is a syringe, a microinfuser, a pen delivery device, or an auto-injector. In one embodiment, the injection device is a single or multi-chamber syringe.

[0042] In yet another embodiment, embodiments of the present disclosure relate to the use of the aqueous pharmaceutical formulations, pharmaceutical dosage forms, sealed containers, pre-filled syringes, pre-filled pens or autoinjectors, or kits disclosed herein in the treatment of diseases or conditions in subjects selected from the group consisting of autoimmune diseases or conditions, inflammatory diseases or conditions, systemic inflammatory diseases or conditions, and transplant rejection diseases or conditions, mediated by hOX40L.

[0043] In yet another embodiment, embodiments of the present disclosure relate to a method for treating a disease or condition in a subject selected from the group consisting of autoimmune diseases or conditions, inflammatory diseases or conditions, systemic inflammatory diseases or conditions, and transplant rejection diseases or conditions, the method comprising administering an effective amount of the aqueous pharmaceutical formulation described herein to a subject in need thereof.

[0044] In a particular embodiment, a method for treating atopic dermatitis is provided, the method comprising administering an effective amount of the aqueous pharmaceutical formulation described herein to a subject in need thereof.

[0045] In a particular embodiment, a method for treating asthma is provided, which comprises administering an effective amount of the aqueous pharmaceutical formulation described herein to a subject in need thereof.

[0046] In another embodiment, an aqueous pharmaceutical formulation is provided comprising (a) an anti-OX40 ligand (OX40L) antagonist antibody or antigen-binding fragment, (b) a stabilizer, (c) a chelating agent, (d) a surfactant, and (e) a buffer comprising histidine or a histidine salt, wherein the pH of the aqueous pharmaceutical formulation is in the range of 5.5 to about 6.5, and the antibody or antigen-binding fragment comprises a VH domain comprising the HCDR1 sequence of SEQ ID NO: 36 or 42, the HCDR2 sequence of SEQ ID NO: 38 or 44, and the HCDR3 sequence of SEQ ID NO: 40 or 46, and a VL domain comprising the LCDR1 sequence of SEQ ID NO: 50 or 56, the LCDR2 sequence of SEQ ID NO: 52 or 58, and the LCDR3 sequence of SEQ ID NO: 54 or 60, wherein the aqueous pharmaceutical formulation is suitable for parenteral administration to mammalian subjects.

[0047] In another embodiment, an aqueous pharmaceutical formulation is provided comprising (a) an anti-OX40 ligand (OX40L) antagonist antibody or antigen-binding fragment, (b) a stabilizer, (c) a surfactant, and (d) a buffer comprising histidine or a histidine salt, wherein the pH of the aqueous pharmaceutical formulation is in the range of 5.5 to about 6.5, and the antibody or antigen-binding fragment comprises a VH domain comprising the HCDR1 sequence of SEQ ID NO: 36 or 42, the HCDR2 sequence of SEQ ID NO: 38 or 44, and the HCDR3 sequence of SEQ ID NO: 40 or 46, and a VL domain comprising the LCDR1 sequence of SEQ ID NO: 50 or 56, the LCDR2 sequence of SEQ ID NO: 52 or 58, and the LCDR3 sequence of SEQ ID NO: 54 or 60, and the aqueous pharmaceutical formulation is suitable for parenteral administration to mammalian subjects.

[0048] In a particular embodiment, the VL domain contains the amino acid sequence of SEQ ID NO: 48, and the VH domain contains the amino acid sequence of SEQ ID NO: 34.

[0049] In a particular embodiment, the antibody comprises a heavy chain and a light chain, the amino acid sequence of the heavy chain being the amino acid sequence of SEQ ID NO: 62, and the amino acid sequence of the light chain being the amino acid sequence of SEQ ID NO: 64.

[0050] The accompanying drawings incorporated herein and constituting part of herein illustrate exemplary embodiments of this disclosure and, together with the general description set forth above and the detailed description set forth below, help to illustrate the features of this disclosure. [Brief explanation of the drawing]

[0051] [Figure 1A] The graphs show the changes in HMWS of the formulations contained in OMPI syringes when stored for 12 weeks at 5°C (Figure 1A), 25°C (Figure 1B), and 40°C (Figure 1C), respectively, as evaluated by SEC-HPLC in the buffer / pH screening study of Example 1. [Figure 1B] The graphs show the changes in HMWS of the formulations contained in OMPI syringes when stored for 12 weeks at 5°C (Figure 1A), 25°C (Figure 1B), and 40°C (Figure 1C), respectively, as evaluated by SEC-HPLC in the buffer / pH screening study of Example 1. [Figure 1C] The graphs show the changes in HMWS of the formulations contained in OMPI syringes when stored for 12 weeks at 5°C (Figure 1A), 25°C (Figure 1B), and 40°C (Figure 1C), respectively, as evaluated by SEC-HPLC in the buffer / pH screening study of Example 1. [Figure 2A] The graphs show the changes in invisible particles larger than 2 μm in the exemplary formulations contained in OMPI syringes, as evaluated by a particle counter in the buffer / pH screening study of Example 1, when stored for 12 weeks at 5°C (Figure 3A), 25°C (Figure 2B), and 40°C (Figure 2C). [Figure 2B] The graphs show the changes in invisible particles larger than 2 μm in the exemplary formulations contained in OMPI syringes, as evaluated by a particle counter in the buffer / pH screening study of Example 1, when stored for 12 weeks at 5°C (Figure 3A), 25°C (Figure 2B), and 40°C (Figure 2C). [Figure 2C]The graphs show the changes in invisible particles larger than 2 μm in the exemplary formulations contained in OMPI syringes, as evaluated by a particle counter in the buffer / pH screening study of Example 1, when stored for 12 weeks at 5°C (Figure 3A), 25°C (Figure 2B), and 40°C (Figure 2C). [Figure 3A] The graphs show the changes in invisible particles larger than 10 μm in the exemplary formulations contained in OMPI syringes, as evaluated by a particle counter in the buffer / pH screening study of Example 1, when stored for 12 weeks at 5°C (Figure 3A), 25°C (Figure 3B), and 40°C (Figure 3C). [Figure 3B] The graphs show the changes in invisible particles larger than 10 μm in the exemplary formulations contained in OMPI syringes, as evaluated by a particle counter in the buffer / pH screening study of Example 1, when stored for 12 weeks at 5°C (Figure 3A), 25°C (Figure 3B), and 40°C (Figure 3C). [Figure 3C] The graphs show the changes in invisible particles larger than 10 μm in the exemplary formulations contained in OMPI syringes, as evaluated by a particle counter in the buffer / pH screening study of Example 1, when stored for 12 weeks at 5°C (Figure 3A), 25°C (Figure 3B), and 40°C (Figure 3C). [Figure 4A] The graphs show the changes in invisible particles larger than 25 μm in the exemplary formulations contained in OMPI syringes, as evaluated by a particle counter in the buffer / pH screening study of Example 1, when stored for 12 weeks at 5°C (Figure 4A), 25°C (Figure 4B), and 40°C (Figure 4C). [Figure 4B] The graphs show the changes in invisible particles larger than 25 μm in the exemplary formulations contained in OMPI syringes, as evaluated by a particle counter in the buffer / pH screening study of Example 1, when stored for 12 weeks at 5°C (Figure 4A), 25°C (Figure 4B), and 40°C (Figure 4C). [Figure 4C]The graphs show the changes in invisible particles larger than 25 μm in the exemplary formulations contained in OMPI syringes, as evaluated by a particle counter in the buffer / pH screening study of Example 1, when stored for 12 weeks at 5°C (Figure 4A), 25°C (Figure 4B), and 40°C (Figure 4C). [Figure 5A] The graphs show the turbidity results of the formulation samples contained in OMPI syringes, as evaluated by a particle counter in the buffer / pH screening study of Example 1, when stored for 12 weeks at 5°C (Figure 5A), 25°C (Figure 5B), and 40°C (Figure 5C). [Figure 5B] The graphs show the turbidity results of the formulation samples contained in OMPI syringes, as evaluated by a particle counter in the buffer / pH screening study of Example 1, when stored for 12 weeks at 5°C (Figure 5A), 25°C (Figure 5B), and 40°C (Figure 5C). [Figure 5C] The graphs show the turbidity results of the formulation samples contained in OMPI syringes, as evaluated by a particle counter in the buffer / pH screening study of Example 1, when stored for 12 weeks at 5°C (Figure 5A), 25°C (Figure 5B), and 40°C (Figure 5C). [Figure 6A] The graphs show the purity and fragmentation of formulation samples contained in OMPI syringes, measured by capillary electrophoresis, after being stored for 12 weeks at 5°C (Figure 6A), 25°C (Figure 6B), and 40°C (Figure 6C), as evaluated in the buffer / pH screening study of Example 1. [Figure 6B] The graphs show the purity and fragmentation of formulation samples contained in OMPI syringes, measured by capillary electrophoresis, after being stored for 12 weeks at 5°C (Figure 6A), 25°C (Figure 6B), and 40°C (Figure 6C), as evaluated in the buffer / pH screening study of Example 1. [Figure 6C]The graphs show the purity and fragmentation of formulation samples contained in OMPI syringes, measured by capillary electrophoresis, after being stored for 12 weeks at 5°C (Figure 6A), 25°C (Figure 6B), and 40°C (Figure 6C), as evaluated in the buffer / pH screening study of Example 1. [Figure 7A] The graph shows the quantification of PS80 for exemplary formulations stored at syringe reference numbers 1-6 and 11-16 over 12 weeks at 5°C (Figure 7A), 25°C (Figure 7B), and 40°C (Figure 7C), as evaluated in the buffer / pH screening study of Example 1. [Figure 7B] The graph shows the quantification of PS80 for exemplary formulations stored at syringe reference numbers 1-6 and 11-16 over 12 weeks at 5°C (Figure 7A), 25°C (Figure 7B), and 40°C (Figure 7C), as evaluated in the buffer / pH screening study of Example 1. [Figure 7C] The graph shows the quantification of PS80 for exemplary formulations stored at syringe reference numbers 1-6 and 11-16 over 12 weeks at 5°C (Figure 7A), 25°C (Figure 7B), and 40°C (Figure 7C), as evaluated in the buffer / pH screening study of Example 1. [Figure 8A] The graph shows the charge variants analyzed by capillary isoelectric focusing (cIEF) for exemplary formulations evaluated in the buffer / pH screening study of Example 1, stored in syringes with reference numbers 1-6 and 11-16 over 8 weeks at 5°C (Figure 8A), 25°C (Figure 8B), and 40°C (Figure 8C). [Figure 8B] The graph shows the charge variants analyzed by capillary isoelectric focusing (cIEF) for exemplary formulations evaluated in the buffer / pH screening study of Example 1, stored in syringes with reference numbers 1-6 and 11-16 over 8 weeks at 5°C (Figure 8A), 25°C (Figure 8B), and 40°C (Figure 8C). [Figure 8C]The graph shows the charge variants analyzed by capillary isoelectric focusing (cIEF) for exemplary formulations evaluated in the buffer / pH screening study of Example 1, stored in syringes with reference numbers 1-6 and 11-16 over 8 weeks at 5°C (Figure 8A), 25°C (Figure 8B), and 40°C (Figure 8C). [Figure 9] The graph shows the deamide of antibodies at HC N332 in exemplary formulations contained in syringes reference numbers 1_s~6_s and 11_s~16_s after 3 months of storage at 40°C, as evaluated in the buffer / pH screening study of Example 1. [Figure 10] The graph shows the oxidation of antibodies at HC M103 in exemplary formulations contained in syringes reference numbers 1_s~6_s and 11_s~16_s after 3 months of storage at 40°C, as evaluated in the buffer / pH screening study of Example 1. [Figure 11] The graph shows the pH measurements of the formulations contained in syringes reference numbers 1_s~6_s and 11_s~16_s, evaluated in the buffer / pH screening study of Example 1, at t0 and after 12 weeks of storage at 25°C and 40°C. [Figure 12] As described in the buffer / pH screening study of Example 1, the osmotic pressure measurements of the formulations contained in syringes reference numbers 1_s~6_s and 11_s~16_s after t0 and 12 weeks of storage at 40°C are shown in the graph. [Figure 13A] As described in the stabilizer study of Example 1, the graph shows the agglutination measurements evaluated by SEC-HPLC for formulations contained in syringe reference numbers 1_s, 7_s, 8_s, 11_s, 17_x, and 18_s over 12 weeks of storage at 5°C (Figure 13A), 25°C (Figure 13B), and 40°C (Figure 13C). [Figure 13B]As described in the stabilizer study of Example 1, the graph shows the agglutination measurements evaluated by SEC-HPLC for formulations contained in syringe reference numbers 1_s, 7_s, 8_s, 11_s, 17_x, and 18_s over 12 weeks of storage at 5°C (Figure 13A), 25°C (Figure 13B), and 40°C (Figure 13C). [Figure 13C] As described in the stabilizer study of Example 1, the graph shows the agglutination measurements evaluated by SEC-HPLC for formulations contained in syringe reference numbers 1_s, 7_s, 8_s, 11_s, 17_x, and 18_s over 12 weeks of storage at 5°C (Figure 13A), 25°C (Figure 13B), and 40°C (Figure 13C). [Figure 14A] As described in the stabilizer study of Example 1, the graph shows the changes in microscopic particles larger than 2 μm in the exemplary formulations contained in syringe reference numbers 1_s, 7_s, 8_s, 11_s, 17_x, and 18_s over 12 weeks of storage at 5°C (Figure 14A), 25°C (Figure 14B), and 40°C (Figure 14C). [Figure 14B] As described in the stabilizer study of Example 1, the graph shows the changes in microscopic particles larger than 2 μm in the exemplary formulations contained in syringe reference numbers 1_s, 7_s, 8_s, 11_s, 17_x, and 18_s over 12 weeks of storage at 5°C (Figure 14A), 25°C (Figure 14B), and 40°C (Figure 14C). [Figure 14C] As described in the stabilizer study of Example 1, the graph shows the changes in microscopic particles larger than 2 μm in the exemplary formulations contained in syringe reference numbers 1_s, 7_s, 8_s, 11_s, 17_x, and 18_s over 12 weeks of storage at 5°C (Figure 14A), 25°C (Figure 14B), and 40°C (Figure 14C). [Figure 15A]As described in the stabilizer study of Example 1, the graph shows the changes in microscopic particles larger than 10 μm in the exemplary formulations contained in syringe reference numbers 1_s, 7_s, 8_s, 11_s, 17_x, and 18_s over 12 weeks of storage at 5°C (Figure 15A), 25°C (Figure 15B), and 40°C (Figure 15C). [Figure 15B] As described in the stabilizer study of Example 1, the graph shows the changes in microscopic particles larger than 10 μm in the exemplary formulations contained in syringe reference numbers 1_s, 7_s, 8_s, 11_s, 17_x, and 18_s over 12 weeks of storage at 5°C (Figure 15A), 25°C (Figure 15B), and 40°C (Figure 15C). [Figure 15C] As described in the stabilizer study of Example 1, the graph shows the changes in microscopic particles larger than 10 μm in the exemplary formulations contained in syringe reference numbers 1_s, 7_s, 8_s, 11_s, 17_x, and 18_s over 12 weeks of storage at 5°C (Figure 15A), 25°C (Figure 15B), and 40°C (Figure 15C). [Figure 16A] As described in the stabilizer study of Example 1, the graph shows the changes in invisible particles larger than 25 μm in the exemplary formulations contained in syringe reference numbers 1_s, 7_s, 8_s, 11_s, 17_x, and 18_s over 12 weeks of storage at 5°C (Figure 16A), 25°C (Figure 16B), and 40°C (Figure 16C). [Figure 16B] As described in the stabilizer study of Example 1, the graph shows the changes in invisible particles larger than 25 μm in the exemplary formulations contained in syringe reference numbers 1_s, 7_s, 8_s, 11_s, 17_x, and 18_s over 12 weeks of storage at 5°C (Figure 16A), 25°C (Figure 16B), and 40°C (Figure 16C). [Figure 16C]As described in the stabilizer study of Example 1, the graph shows the changes in invisible particles larger than 25 μm in the exemplary formulations contained in syringe reference numbers 1_s, 7_s, 8_s, 11_s, 17_x, and 18_s over 12 weeks of storage at 5°C (Figure 16A), 25°C (Figure 16B), and 40°C (Figure 16C). [Figure 17A] As described in the stabilizer study of Example 1, the turbidity measurements of exemplary formulations contained in syringe reference numbers 1_s, 7_s, 8_s, 11_s, 17_x, and 18_s after 12 weeks of storage at T0 and 5°C (Figure 17A), 25°C (Figure 17B), and 40°C (Figure 17C) are shown graphically. [Figure 17B] As described in the stabilizer study of Example 1, the turbidity measurements of exemplary formulations contained in syringe reference numbers 1_s, 7_s, 8_s, 11_s, 17_x, and 18_s after 12 weeks of storage at T0 and 5°C (Figure 17A), 25°C (Figure 17B), and 40°C (Figure 17C) are shown graphically. [Figure 17C] As described in the stabilizer study of Example 1, the turbidity measurements of exemplary formulations contained in syringe reference numbers 1_s, 7_s, 8_s, 11_s, 17_x, and 18_s after 12 weeks of storage at T0 and 5°C (Figure 17A), 25°C (Figure 17B), and 40°C (Figure 17C) are shown graphically. [Figure 18A] As described in the stabilizer study of Example 1, the PS80 quantification of exemplary formulations contained in syringe reference numbers 1_s, 7_s, 8_s, 11_s, 17_x, and 18_s after 12 weeks of storage at 5°C (Figure 18A), 25°C (Figure 18B), and 40°C (Figure 18C) is shown graphically. [Figure 18B] As described in the stabilizer study of Example 1, the PS80 quantification of exemplary formulations contained in syringe reference numbers 1_s, 7_s, 8_s, 11_s, 17_x, and 18_s after 12 weeks of storage at 5°C (Figure 18A), 25°C (Figure 18B), and 40°C (Figure 18C) is shown graphically. [Figure 18C]As described in the stabilizer study of Example 1, the PS80 quantification of exemplary formulations contained in syringe reference numbers 1_s, 7_s, 8_s, 11_s, 17_x, and 18_s after 12 weeks of storage at 5°C (Figure 18A), 25°C (Figure 18B), and 40°C (Figure 18C) is shown graphically. [Figure 19] As described in the stabilizer study of Example 1, the deamidation of antibodies at HC N332 in exemplary formulations contained in syringes reference numbers 1_s, 7_s, 8_s, 11_s, 17_x, and 18_s after 3 months of storage at 40°C is shown graphically. [Figure 20] As described in the stabilizer study of Example 1, the oxidation of antibodies at HC M103 in exemplary formulations contained in syringe reference numbers 1_s, 7_s, 8_s, 11_s, 17_x, and 18_s after 3 months of storage at 40°C is graphically shown. [Figure 21A] As described in the syringe comparability study of Example 1, the changes in HMWS of the formulations in syringes 1_s_Omp, 9_s_BD, 10_s_BD, and 11_s_Omp over 12 weeks of storage at 5°C (Figure 21A), 25°C (Figure 21B), and 40°C (Figure 21C) are shown graphically. [Figure 21B] As described in the syringe comparability study of Example 1, the changes in HMWS of the formulations in syringes 1_s_Omp, 9_s_BD, 10_s_BD, and 11_s_Omp over 12 weeks of storage at 5°C (Figure 21A), 25°C (Figure 21B), and 40°C (Figure 21C) are shown graphically. [Figure 21C] As described in the syringe comparability study of Example 1, the changes in HMWS of the formulations in syringes 1_s_Omp, 9_s_BD, 10_s_BD, and 11_s_Omp over 12 weeks of storage at 5°C (Figure 21A), 25°C (Figure 21B), and 40°C (Figure 21C) are shown graphically. [Figure 22A]As described in the syringe comparability study of Example 1, the graphs show the concentrations of microscopic particles larger than 2 μm, as measured by HIAC, in the exemplary formulations contained in syringes 1_s_Omp, 9_s_BD, 10_s_BD, and 11_s_Omp, respectively, over 12 weeks of storage at 5°C (Figure 22A), 25°C (Figure 22B), and 40°C (Figure 22C). [Figure 22B] As described in the syringe comparability study of Example 1, the graphs show the concentrations of microscopic particles larger than 2 μm, as measured by HIAC, in the exemplary formulations contained in syringes 1_s_Omp, 9_s_BD, 10_s_BD, and 11_s_Omp, respectively, over 12 weeks of storage at 5°C (Figure 22A), 25°C (Figure 22B), and 40°C (Figure 22C). [Figure 22C] As described in the syringe comparability study of Example 1, the graphs show the concentrations of microscopic particles larger than 2 μm, as measured by HIAC, in the exemplary formulations contained in syringes 1_s_Omp, 9_s_BD, 10_s_BD, and 11_s_Omp, respectively, over 12 weeks of storage at 5°C (Figure 22A), 25°C (Figure 22B), and 40°C (Figure 22C). [Figure 23A] As described in the syringe comparability study of Example 1, the graphs show the concentrations of microscopic particles larger than 10 μm, as measured by HIAC, in the exemplary formulations contained in syringes 1_s_Omp, 9_s_BD, 10_s_BD, and 11_s_Omp, respectively, over 12 weeks of storage at 5°C (Figure 23A), 25°C (Figure 23B), and 40°C (Figure 23C). [Figure 23B] As described in the syringe comparability study of Example 1, the graphs show the concentrations of microscopic particles larger than 10 μm, as measured by HIAC, in the exemplary formulations contained in syringes 1_s_Omp, 9_s_BD, 10_s_BD, and 11_s_Omp, respectively, over 12 weeks of storage at 5°C (Figure 23A), 25°C (Figure 23B), and 40°C (Figure 23C). [Figure 23C]As described in the syringe comparability study of Example 1, the graphs show the concentrations of microscopic particles larger than 10 μm, as measured by HIAC, in the exemplary formulations contained in syringes 1_s_Omp, 9_s_BD, 10_s_BD, and 11_s_Omp, respectively, over 12 weeks of storage at 5°C (Figure 23A), 25°C (Figure 23B), and 40°C (Figure 23C). [Figure 24A] As described in the syringe comparability study of Example 1, the graphs show the concentrations of invisible particles larger than 25 μm, as measured by HIAC, in the exemplary formulations contained in syringes 1_s_Omp, 9_s_BD, 10_s_BD, and 11_s_Omp, respectively, over 12 weeks of storage at 5°C (Figure 24A), 25°C (Figure 24B), and 40°C (Figure 24C). [Figure 24B] As described in the syringe comparability study of Example 1, the graphs show the concentrations of invisible particles larger than 25 μm, as measured by HIAC, in the exemplary formulations contained in syringes 1_s_Omp, 9_s_BD, 10_s_BD, and 11_s_Omp, respectively, over 12 weeks of storage at 5°C (Figure 24A), 25°C (Figure 24B), and 40°C (Figure 24C). [Figure 24C] As described in the syringe comparability study of Example 1, the graphs show the concentrations of invisible particles larger than 25 μm, as measured by HIAC, in the exemplary formulations contained in syringes 1_s_Omp, 9_s_BD, 10_s_BD, and 11_s_Omp, respectively, over 12 weeks of storage at 5°C (Figure 24A), 25°C (Figure 24B), and 40°C (Figure 24C). [Figure 25A] As described in the syringe comparability study of Example 1, the turbidity measurements for the exemplary formulations contained in syringes 1_s_Omp, 9_s_BD, 10_s_BD, and 11_s_Omp after 12 weeks of storage at 5°C (Figure 25A), 25°C (Figure 25B), and 40°C (Figure 25C), respectively, are shown graphically. [Figure 25B]As described in the syringe comparability study of Example 1, the turbidity measurements for the exemplary formulations contained in syringes 1_s_Omp, 9_s_BD, 10_s_BD, and 11_s_Omp after 12 weeks of storage at 5°C (Figure 25A), 25°C (Figure 25B), and 40°C (Figure 25C), respectively, are shown graphically. [Figure 25C] As described in the syringe comparability study of Example 1, the turbidity measurements for the exemplary formulations contained in syringes 1_s_Omp, 9_s_BD, 10_s_BD, and 11_s_Omp after 12 weeks of storage at 5°C (Figure 25A), 25°C (Figure 25B), and 40°C (Figure 25C), respectively, are shown graphically. [Figure 26A] As described in the syringe comparability study of Example 1, the charge variants of antibodies measured by cIEF in exemplary formulations contained in syringes 1_s_Omp, 9_s_BD, 10_s_BD, and 11_s_Omp at t) over 12 weeks of storage at 5°C (Figure 26A), 25°C (Figure 26B), and 40°C (Figure 26C), respectively, are shown graphically. [Figure 26B] As described in the syringe comparability study of Example 1, the charge variants of antibodies measured by cIEF in exemplary formulations contained in syringes 1_s_Omp, 9_s_BD, 10_s_BD, and 11_s_Omp at t) over 12 weeks of storage at 5°C (Figure 26A), 25°C (Figure 26B), and 40°C (Figure 26C), respectively, are shown graphically. [Figure 26C] As described in the syringe comparability study of Example 1, the charge variants of antibodies measured by cIEF in exemplary formulations contained in syringes 1_s_Omp, 9_s_BD, 10_s_BD, and 11_s_Omp at t) over 12 weeks of storage at 5°C (Figure 26A), 25°C (Figure 26B), and 40°C (Figure 26C), respectively, are shown graphically. [Figure 27A]As described in the chelating agent justification study of Example 1, the graph shows the change in HMWS% measured by SEC-HPLC in sample formulations in syringes with reference numbers C-1 to C-6 over 12 weeks of storage at 5°C (Figure 27A), 25°C (Figure 27B), and 40°C (Figure 27C). [Figure 27B] As described in the chelating agent justification study of Example 1, the graph shows the change in HMWS% measured by SEC-HPLC in sample formulations in syringes with reference numbers C-1 to C-6 over 12 weeks of storage at 5°C (Figure 27A), 25°C (Figure 27B), and 40°C (Figure 27C). [Figure 27C] As described in the chelating agent justification study of Example 1, the graph shows the change in HMWS% measured by SEC-HPLC in sample formulations in syringes with reference numbers C-1 to C-6 over 12 weeks of storage at 5°C (Figure 27A), 25°C (Figure 27B), and 40°C (Figure 27C). [Figure 28A] As described in the chelating agent justification study of Example 1, the PS80 measurements analyzed by CAD in formulations with reference numbers C_1 to C_6 ​​over a two-month storage period at 5°C, 25°C, and 40°C are shown in the graph. [Figure 28B] As described in the chelating agent justification study of Example 1, the graphs show the changes in PS80 in formulations with reference numbers C_1 to C_6 ​​over a 3-month storage period at 5°C (Figure 29A), 25°C (Figure 29B), and 40°C (Figure 29C) (dotted lines indicate method variability). [Figure 28C] As described in the chelating agent justification study of Example 1, the graphs show the changes in PS80 in formulations with reference numbers C_1 to C_6 ​​over a 3-month storage period at 5°C (Figure 29A), 25°C (Figure 29B), and 40°C (Figure 29C) (dotted lines indicate method variability). [Figure 28D]As described in the chelating agent justification study of Example 1, the graphs show the changes in PS80 in formulations with reference numbers C_1 to C_6 ​​over a 3-month storage period at 5°C (Figure 29A), 25°C (Figure 29B), and 40°C (Figure 29C) (dotted lines indicate method variability). [Figure 29] As described in the chelating agent justification study of Example 1, the following shows the weekly percentage change in HC N332 deamidation of antibodies in formulations with reference numbers C1 to C6 over 8 weeks of storage at 40°C. [Figure 30] As described in the chelating agent justification study of Example 1, the weekly percentage change in HC M103 oxidation of antibodies in the formulations over 8 weeks of storage at 40°C is shown graphically. Charge variants were analyzed by CIEF at T0, T2wk (2 weeks), T4wk (4 weeks), and T8wk (8 weeks) for formulations with reference numbers C-1 to C_6, which were stored over 8 weeks at 5°C, 25°C, and 40°C, as described in the chelating agent justification study of Example 1. [Figure 31A] As described in the chelating agent justification study of Example 1, the charge variants measured by cIEF for antibodies in formulations with reference numbers C_1 to C_6 ​​over 8 weeks of storage at 5°C, 25°C, and 40°C are shown graphically, with acidic peaks shown in Figure 31A, basic peaks in Figure 31B, and major peaks in Figure 31C. [Figure 31B] As described in the chelating agent justification study of Example 1, the charge variants measured by cIEF for antibodies in formulations with reference numbers C_1 to C_6 ​​over 8 weeks of storage at 5°C, 25°C, and 40°C are shown graphically, with acidic peaks shown in Figure 31A, basic peaks in Figure 31B, and major peaks in Figure 31C. [Figure 31C] As described in the chelating agent justification study of Example 1, the charge variants measured by cIEF for antibodies in formulations with reference numbers C_1 to C_6 ​​over 8 weeks of storage at 5°C, 25°C, and 40°C are shown graphically, with acidic peaks shown in Figure 31A, basic peaks in Figure 31B, and major peaks in Figure 31C. [Figure 32A] This graph shows the measured changes in HMWS for formulations containing 31 mg / mL and 125 mg / mL of antibody at time points T0, T1hr, T3hr, and T6hr over the list action study period, as described in the PS80 justification study of Example 1. [Figure 32B] This graph shows the measured changes in HMWS for formulations containing 31 mg / mL and 125 mg / mL of antibody at time points T0, T1hr, T3hr, and T6hr over the list action study period, as described in the PS80 justification study of Example 1. [Figure 32C] This graph shows the measured changes in HMWS for formulations containing 31 mg / mL and 125 mg / mL of antibody at time points T0, T1hr, T3hr, and T6hr over the list action study period, as described in the PS80 justification study of Example 1. [Figure 33A] As described in the PS80 justification study of Example 1, the turbidity analysis for exemplary formulations with 31 mg / mL and 125 mg / mL of antibody at time points T0, T1hr, T3hr, and T6hr over the list action study period is shown graphically. [Figure 33B] As described in the PS80 justification study of Example 1, the turbidity analysis for exemplary formulations with 31 mg / mL and 125 mg / mL of antibody at time points T0, T1hr, T3hr, and T6hr over the list action study period is shown graphically. [Figure 33C] As described in the PS80 justification study of Example 1, the turbidity analysis for exemplary formulations with 31 mg / mL and 125 mg / mL of antibody at time points T0, T1hr, T3hr, and T6hr over the list action study period is shown graphically. [Figure 34A]As described in the PS80 justification study of Example 1, the graph shows the number of invisible particles of at least 2 μm, at least 10 μm, or at least 25 μm measured by HIAC in the exemplary formulations having antibody concentrations of 31 mg / mL and 125 mg / mL at time points T0, T1hr, T3hr, and T6hr over the list action study period. [Figure 34B] As described in the PS80 justification study of Example 1, the graph shows the number of invisible particles of at least 2 μm, at least 10 μm, or at least 25 μm measured by HIAC in the exemplary formulations having antibody concentrations of 31 mg / mL and 125 mg / mL at time points T0, T1hr, T3hr, and T6hr over the list action study period. [Figure 34C] As described in the PS80 justification study of Example 1, the graph shows the number of invisible particles of at least 2 μm, at least 10 μm, or at least 25 μm measured by HIAC in the exemplary formulations having antibody concentrations of 31 mg / mL and 125 mg / mL at time points T0, T1hr, T3hr, and T6hr over the list action study period. [Figure 34D] As described in the PS80 justification study of Example 1, the graph shows the number of invisible particles of at least 2 μm, at least 10 μm, or at least 25 μm measured by HIAC in the exemplary formulations having antibody concentrations of 31 mg / mL and 125 mg / mL at time points T0, T1hr, T3hr, and T6hr over the list action study period. [Figure 34E] As described in the PS80 justification study of Example 1, the graph shows the number of invisible particles of at least 2 μm, at least 10 μm, or at least 25 μm measured by HIAC in the exemplary formulations having antibody concentrations of 31 mg / mL and 125 mg / mL at time points T0, T1hr, T3hr, and T6hr over the list action study period. [Figure 34F] As described in the PS80 justification study of Example 1, the graph shows the number of invisible particles of at least 2 μm, at least 10 μm, or at least 25 μm measured by HIAC in the exemplary formulations having antibody concentrations of 31 mg / mL and 125 mg / mL at time points T0, T1hr, T3hr, and T6hr over the list action study period. [Figure 35A] As described in the PS80 justification study of Example 1, the graph shows the results of HMWS changes analyzed by SEC-HPLC for 31 mg / mL and 125 mg / mL formulations over the orbital shaking study period. [Figure 35B] As described in the PS80 justification study of Example 1, the graph shows the results of HMWS changes analyzed by SEC-HPLC for 31 mg / mL and 125 mg / mL formulations over the orbital shaking study period. [Figure 36A] As described in the PS80 justification study of Example 1, for exemplary formulations having 31 mg / mL and 125 mg / mL of antibody at time points T0, T1hr, T3hr, and T6hr over the orbital shaking study period, the graph shows microscopic particles of at least 2 μm, at least 10 μm, or at least 25 μm as measured by HIAC. [Figure 36B] As described in the PS80 justification study of Example 1, for exemplary formulations having 31 mg / mL and 125 mg / mL of antibody at time points T0, T1hr, T3hr, and T6hr over the orbital shaking study period, the graph shows microscopic particles of at least 2 μm, at least 10 μm, or at least 25 μm as measured by HIAC. [Figure 36C]As described in the PS80 justification study of Example 1, for exemplary formulations having 31 mg / mL and 125 mg / mL of antibody at time points T0, T1hr, T3hr, and T6hr over the orbital shaking study period, the graph shows microscopic particles of at least 2 μm, at least 10 μm, or at least 25 μm as measured by HIAC. [Figure 36D] As described in the PS80 justification study of Example 1, for exemplary formulations having 31 mg / mL and 125 mg / mL of antibody at time points T0, T1hr, T3hr, and T6hr over the orbital shaking study period, the graph shows microscopic particles of at least 2 μm, at least 10 μm, or at least 25 μm as measured by HIAC. [Figure 36E] As described in the PS80 justification study of Example 1, for exemplary formulations having 31 mg / mL and 125 mg / mL of antibody at time points T0, T1hr, T3hr, and T6hr over the orbital shaking study period, the graph shows microscopic particles of at least 2 μm, at least 10 μm, or at least 25 μm as measured by HIAC. [Figure 36F] As described in the PS80 justification study of Example 1, for exemplary formulations having 31 mg / mL and 125 mg / mL of antibody at time points T0, T1hr, T3hr, and T6hr over the orbital shaking study period, the graph shows microscopic particles of at least 2 μm, at least 10 μm, or at least 25 μm as measured by HIAC. [Figure 37A] As described in the PS80 justification study of Example 1, the turbidity for 31 mg / mL and 125 mg / mL formulations over the study period using list action and silicone oil is shown graphically. [Figure 37B]As described in the PS80 justification study of Example 1, the turbidity for 31 mg / mL and 125 mg / mL formulations over the study period using list action and silicone oil is shown graphically. [Figure 38] As described in Example 2, the changes in HMWS analyzed by SEC-HPLC for formulations F2-1, F2-2, and F2-3 over a 3-month storage period at 5°C, 25°C, and 40°C are shown in the graph. [Figure 39] As detailed in Example 2, the PS80 loss of formulations F2-1, F2-2, and F2-3 over 3 months of storage at 5°C, 25°C, and 40°C is shown graphically. [Figure 40] The graph shows the weekly percentage change in deamidation of antibodies in HC D332 in formulations F2-1, F2-2, and F2-3 after 1 month of storage at 40°C, as measured by peptide mapping in Example 2. [Figure 41] As detailed in Example 2, the graph shows the percentage change in weekly oxidation of antibodies in HC M103 in formulations F2-1, F2-2, and F2-3 after 1 month of storage at 40°C, as measured by peptide mapping. [Figure 42] As detailed in Example 2, this graph shows the charge variants measured by capillary isoelectric focusing (cIEF) for formulations F2-1, F2-2, and F2-3 over a two-month storage period at 5°C, 25°C, and 40°C. [Figure 43] This table shows the stability results of formulations F3-1, F3-2, and F3-3 in containers, analyzed over a 3-month storage period at 5°C, 25°C, and 40°C, according to the study in Example 3. [Figure 44] The results of the gliding force of 2.23 mL of BD and OMPI PFS, as in the study of Example 4, are shown in a graph. [Figure 45]The viscoelastic behavior of amriterimab at a concentration of 263 mg / ml in 25 mM histidine / histidine hydrochloride buffer (HisHCl) at pH 6.2, analyzed by shear rate ramping at 25°C from 0 to 4000 s⁻¹, as described in Example 5, is shown in the graph. [Figure 46] As described in Example 5, the concentration-dependent viscosity profiles of amriterimab in 25 mM histidine / histidine hydrochloride buffer (HisHCl) at pH 6.2 at 5°C and 25°C are shown graphically. [Figure 47] The graph shows the protein content of the formulation samples measured by UV / Vis at the initial time point T0, and after exposure to freeze-thaw stress (-65°C to 25°C) or shaking stress at ambient temperature and low temperatures. [Figure 48] The graph shows the pH values ​​of the formulation samples at the initial time point T0, and after exposure to freeze-thaw stress (-65°C to 25°C) or shaking stress at ambient temperature and low temperatures. [Figure 49] The graph shows the surfactant content of the formulation samples at the initial time point T0, and after exposure to freeze-thaw stress (-65°C to 25°C) or shaking stress at ambient temperature and low temperatures. [Figure 50] The graph shows the clarity and opacity (turbidity) of the formulation sample solution at the initial time point T0, and after exposure to freeze-thaw stress (-65°C to 25°C) or shaking stress at ambient temperature and low temperatures. [Figure 51A] Figure 51A shows the percentage of major peaks and low molecular weight species (LMWS) for all formulation samples subjected to freeze-thaw stress (-65°C to 25°C) or shaking stress at ambient temperature and low temperatures, as analyzed by size exclusion chromatography (SE-HPLC) in Example 6. The graphs show the differences in the percentages of major peaks obtained from SE-HPLC analysis, Figure 51B shows the percentage of HMW species obtained from SE-HPLC analysis, and Figure 51C shows the percentage of HMW species obtained from SE-HPLC analysis. [Figure 51B]Figure 51A shows the percentage of major peaks and low molecular weight species (LMWS) for all formulation samples subjected to freeze-thaw stress (-65°C to 25°C) or shaking stress at ambient temperature and low temperatures, as analyzed by size exclusion chromatography (SE-HPLC) in Example 6. The graphs show the differences in the percentages of major peaks obtained from SE-HPLC analysis, Figure 51B shows the percentage of HMW species obtained from SE-HPLC analysis, and Figure 51C shows the percentage of HMW species obtained from SE-HPLC analysis. [Figure 51C] Figure 51A shows the percentage of major peaks and low molecular weight species (LMWS) for all formulation samples subjected to freeze-thaw stress (-65°C to 25°C) or shaking stress at ambient temperature and low temperatures, as analyzed by size exclusion chromatography (SE-HPLC) in Example 6. The graphs show the differences in the percentages of major peaks obtained from SE-HPLC analysis, Figure 51B shows the percentage of HMW species obtained from SE-HPLC analysis, and Figure 51C shows the percentage of HMW species obtained from SE-HPLC analysis. [Figure 52A] The graphs in Example 2 show the number of invisible particles in all formulation samples analyzed by HIAC. Figure 52A includes panels (a) and (b) which graphically show the count of invisible particles having a size of at least 2 μm for all formulation samples, with panel (a) showing counts in the range of 0 to 12000 (counts / mL) and panel (b) showing counts in the range of 0 to 2000 (counts / mL). Figure 52B includes panels (a), (b), and (c), which graphically show the count of invisible particles having a size of at least 5 μm (panel a), at least 10 (panel b), and at least 25 (panel c), respectively, for all formulation samples. [Figure 52B]The graphs in Example 2 show the number of invisible particles in all formulation samples analyzed by HIAC. Figure 52A includes panels (a) and (b) which graphically show the count of invisible particles having a size of at least 2 μm for all formulation samples, with panel (a) showing counts in the range of 0 to 12000 (counts / mL) and panel (b) showing counts in the range of 0 to 2000 (counts / mL). Figure 52B includes panels (a), (b), and (c), which graphically show the count of invisible particles having a size of at least 5 μm (panel a), at least 10 (panel b), and at least 25 (panel c), respectively, for all formulation samples. [Figure 53A] The graphs show the evaluation of charged variants for all formulation samples stored for 8 or 16 weeks at -65°C, 5°C, 25°C, and 40°C, or subjected to freeze-thaw stress (-65°C to 25°C) or shaking stress at ambient temperature and low temperatures. Figure 53A shows the graph of the major protein peaks measured by imaging capillary isoelectric focusing (iCIEF). Figure 53B shows the graph of acidic variants measured by imaging capillary isoelectric focusing (iCIEF). Figure 53C shows the graph of basic variants measured by imaging capillary isoelectric focusing (iCIEF). [Figure 53B] The graphs show the evaluation of charged variants for all formulation samples stored for 8 or 16 weeks at -65°C, 5°C, 25°C, and 40°C, or subjected to freeze-thaw stress (-65°C to 25°C) or shaking stress at ambient temperature and low temperatures. Figure 53A shows the graph of the major protein peaks measured by imaging capillary isoelectric focusing (iCIEF). Figure 53B shows the graph of acidic variants measured by imaging capillary isoelectric focusing (iCIEF). Figure 53C shows the graph of basic variants measured by imaging capillary isoelectric focusing (iCIEF). [Figure 53C]The graphs show the evaluation of charged variants for all formulation samples stored for 8 or 16 weeks at -65°C, 5°C, 25°C, and 40°C, or subjected to freeze-thaw stress (-65°C to 25°C) or shaking stress at ambient temperature and low temperatures. Figure 53A shows the graph of the major protein peaks measured by imaging capillary isoelectric focusing (iCIEF). Figure 53B shows the graph of acidic variants measured by imaging capillary isoelectric focusing (iCIEF). Figure 53C shows the graph of basic variants measured by imaging capillary isoelectric focusing (iCIEF). [Modes for carrying out the invention]

[0052] Embodiments of this disclosure provide aqueous pharmaceutical formulations comprising an anti-OX40L antibody or its antigen-binding fragment for treating or preventing OX40L-mediated diseases or conditions in humans. Embodiments also provide packaged pharmaceuticals comprising a container containing the formulations disclosed herein. In some embodiments, the anti-OX40L antibody is amliterimab, an amliterimab variant, or its antigen-binding fragment.

[0053] Before describing this disclosure in detail, it should be understood that the embodiments described herein are not limited to specific methods and experimental conditions, as such methods and conditions may vary. It should also be understood that the terms used herein are intended solely to describe specific embodiments and are not intended to limit their scope. Except in the examples of operations, or where otherwise indicated, all figures representing quantities of components or reaction conditions used herein should be understood to disclose those figures, and in all cases modified by the term “approximately.”

[0054] definition The following are definitions of various terms used to describe the embodiments disclosed herein. These definitions apply to the terms as used throughout this specification and the claims, either individually or as part of a larger group, unless specifically limited in particular examples.

[0055] Unless otherwise defined, all scientific and technical terms used herein have meanings that are generally understood by those skilled in the art. Furthermore, unless otherwise required by context, singular forms shall include plural forms and plural forms shall include singular forms. In general, the terminology and techniques used herein in connection with cell and tissue culture, molecular biology, and the chemistry and hybridization of proteins and oligonucleotides or polynucleotides are well known and commonly used in the art.

[0056] As used herein and in the appended claims, the singular forms “a,” “an,” and “the” include multiple references unless otherwise explicitly indicated by their content. Thus, for example, a reference to “molecule” includes, optionally, a combination of two or more such molecules. The use of “or” herein is inclusive.

[0057] Where used herein, the terms “about” or “approximately” are to be understood by those skilled in the art and vary to some extent depending on the context in which they are used. When referring to measurable values ​​such as quantities or temporal durations, the terms “about” or “approximately” mean to include variations of ±20%, ±15%, or ±10%, including ±5%, ±1%, and ±0.1% from a given value, where such variations are appropriate for carrying out the disclosed method.

[0058] As used herein and in the claims, the terms “comprising” (and any form of “comprise,” such as “comprise” and “comprises”), “having” (and any form of “have,” such as “have” and “has”), “including” (and any form of “includes,” such as “includes” and “include”), or “containing” (and any form of “contains,” such as “contains” and “contain”) are comprehensive or open-ended and do not exclude additional, unenumerated elements or method steps.

[0059] As used herein, the term “essentially derived from” refers to elements necessary for a given embodiment. This term allows for the presence of elements that do not substantially affect the basic, novel, or functional characteristics of that embodiment.

[0060] The abbreviation "e.g." originates from the Latin "exempli gratia" and is used herein to indicate non-restrictive examples. Therefore, the abbreviation "e.g." is synonymous with the term "for example."

[0061] As used herein, the term “amino acid” includes alanine (Ala or A), arginine (Arg or R), asparagine (Asn or N), aspartic acid (Asp or D), cysteine ​​(Cys or C), glutamine (Gin or Q), glutamic acid (Glu or E), glycine (Gly or G), histidine (His or H), isoleucine (lie or I), leucine (Leu or L), lysine (Lys or K), methionine (Met or M), phenylalanine (Phe or F), proline (Pro or P), serine (Ser or S), threonine (Thr or T), tryptophan (Trp or W), tyrosine (Tyr or Y), and valine (Val or V). Unconventional amino acids are also within the scope of this disclosure and include norleucine, ornithine, norvaline, homoserine, and other amino acid residue analogs, such as those described in Ellman et al. Meth. Enzymol. 202:301-336 (1991). To generate such naturally occurring amino acid residues, the procedures of Noren et al. Science 244:182 (1989) and Ellman et al., (above) can be used. Briefly, these procedures involve chemically activating a suppressor tRNA having naturally occurring amino acid residues, followed by in vitro transcription and RNA translation. The introduction of unconventional amino acids can also be achieved using peptide chemistry known in the art. As used herein, the term “polar amino acid” includes amino acids that have a net zero charge but have non-zero partial charges in different parts of their side chains (e.g., M, F, W, S, Y, N, Q, C). These amino acids can be involved in hydrophobic and electrostatic interactions. As used herein, the term “charged amino acid” includes amino acids that may have a non-zero net charge on their side chains (e.g., R, K, H, E, D). These amino acids may be involved in hydrophobic and electrostatic interactions.

[0062] As used herein, the term “conservative amino acid substitution” refers to an amino acid substitution resulting from replacing one amino acid with another amino acid having similar structural and / or chemical properties, such as replacing leucine with isoleucine or valine, replacing aspartate with glutamate, or replacing threonine with serine. Therefore, a “conservative substitution” of a particular amino acid sequence refers to a substitution of an amino acid that is not important to polypeptide activity, or an amino acid substitution with another amino acid having similar properties (e.g., acidic, basic, positive or negative charge, polar or nonpolar), so that even if the amino acid is important, the substitution does not reduce the peptide's activity (i.e., its ability to cross the blood-brain barrier (BBB)). Tables of conservative substitutions resulting in functionally similar amino acids are well known in the art. For example, the following six groups each contain amino acids that are conserved substitutions with respect to each other: 1) alanine (A), serine (S), threonine (T); 2) aspartic acid (D), glutamic acid (E); 3) asparagine (N), glutamine (Q); 4) arginine (R), lysine (K); 5) isoleucine (I), leucine (L), methionine (M), valine (V); and 6) phenylalanine (F), tyrosine (Y), tryptophan (W). (See also Creighton, Proteins, WH Freeman and Company (1984), which incorporates the entire group by reference). In some embodiments, individual substitutions, deletions, or additions that modify, add, or delete one or a small number of amino acids may also be considered “conservative substitutions” if the change does not reduce the activity of the peptide. Insertions or deletions typically range from about one to five amino acids. The selection of a conserved amino acid may be based on the position of the amino acid to be substituted in the peptide, for example, whether the amino acid is outside the peptide and exposed to the solvent, or inside the peptide and not exposed to the solvent.

[0063] In alternative embodiments, an amino acid to substitute for an existing amino acid can be selected based on the position of the existing amino acid, i.e., its exposure to the solvent (i.e., whether the amino acid is exposed to the solvent or located on the outer surface of the peptide or polypeptide, compared to internally localized amino acids that are not exposed to the solvent). Such selection of conserved amino acid substitutions is well known in the art, as disclosed, for example, in Dordo et al., J. Mol Biol, 1999, 217, 721-739 and Taylor et al., J. Theor. Biol. 119(1986); 205-218 and S. French and B. Robson, J. Mol. Evol., 19(1983) 171. Therefore, suitable conserved amino acid substitutions can be selected for the amino acids outside the protein or peptide (i.e., amino acids exposed to the solvent), for example, the following substitutions may be used, but are not limited to: substitution of Y with F, substitution of T with S or K, substitution of P with A, substitution of E with D or Q, substitution of N with D or G, substitution of R with K, substitution of G with N or A, substitution of T with S or K, substitution of D with N or E, substitution of I with L or V, substitution of F with Y, substitution of S with T or A, substitution of R with K, substitution of G with N or A, substitution of K with R, and substitution of A with S, K, or P.

[0064] In alternative embodiments, suitable conserved amino acid substitutions can be selected for amino acids inside a protein or peptide, for example, suitable conserved substitutions for amino acids inside a protein or peptide (i.e., amino acids not exposed to a solvent) can be used, for example, the following conserved substitutions can be used: substitution of Y with F, substitution of T with A or S, substitution of I with L or V, substitution of W with Y, substitution of M with L, substitution of N with D, substitution of G with A, substitution of T with A or S, substitution of D with N, substitution of I with L or V, substitution of F with Y or L, substitution of S with A or T, and substitution of A with S, G, T, or V. In some embodiments, non-conserved amino acid substitutions are also included in the terminology of the variant.

[0065] As used herein, the term “administer” means, in the course of this disclosure, distributing, delivering, or applying an active compound, i.e., an antibody or its antigen-binding fragment, to a subject in a pharmaceutical formulation by any preferred route for delivering the active compound to the subject. Examples of routes of administration include, but are not limited to, subcutaneous, intravenous, e.g., intravenous injection and intravenous infusion (e.g., via central venous access, intramuscular, oral, nasal, and intrapulmonary administration).

[0066] As used herein, the term “antibody” generally refers to immunoglobulin molecules, as well as their polymers (e.g., IgM), which consist of four polypeptide chains: two heavy (H) chains and two light (L) chains interconnected by disulfide bonds. However, immunoglobulin molecules consisting only of heavy chains (i.e., lacking light chains) are also included in the definition of “antibody.” Each heavy chain contains a heavy chain variable region (hereinafter abbreviated as HCVR or VH) and a heavy chain constant region. The heavy chain constant region contains three domains: CH1, CH2, and CH3. Each light chain contains a light chain variable region (hereinafter abbreviated as LCVR or VL) and a light chain constant region. The light chain constant region contains one domain (CL1). The VH and VL regions can be further subdivided into more conserved regions called framework regions (FR) and scattered, highly variable regions called complementarity-determining regions (CDR). Each VH and VL consists of three CDRs and four FRs arranged from the amino terminus to the carboxyl terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.

[0067] As used herein, the terms “antagonistic antibody” or “antagonist antibody” are used herein by equivalent and include, for example, antibodies that can inhibit and / or neutralize the biological signaling activity of OX40 by blocking or substantially reducing the binding of OX40 to the OX40 ligand (OX40L), and thus inhibit or reduce OX40-induced signaling pathways and / or inhibit or reduce OX40-mediated cellular responses such as lymphocyte proliferation, cytokine expression, or lymphocyte survival. Such antibodies may include, for example, OX40L antagonist antibodies or OX40L antagonist antibodies or OX40 antagonist antibodies or OX40 antagonist antibodies.

[0068] Unless otherwise specifically indicated, the term “antibody” as used herein should be understood to include the complete antibody molecule as well as its antigen-binding fragments. As used herein, the terms “antigen-binding moiety” or “antibody-binding fragment” (or simply “antibody moiety” or “antibody fragment”) refer to one or more fragments of an antibody that possess the ability to specifically bind to a target antigen, such as human OX40L or its epitope.

[0069] As used herein, the term “antibody fragment” refers to a polypeptide comprising at least one immunoglobulin variable domain or immunoglobulin variable domain sequence that specifically binds to a given antigen. An antibody fragment may include an antibody or a polypeptide comprising the antigen-binding domain of an antibody. In some embodiments, an antibody fragment may include a monoclonal antibody or a polypeptide comprising the antigen-binding domain of a monoclonal antibody. For example, an antibody may include a heavy (H) chain variable region (abbreviated herein as VH) and a light (L) chain variable region (abbreviated herein as VL). In another example, an antibody may include two heavy (H) chain variable regions and two light (L) chain variable regions. The term "antibody fragment" encompasses antigen-binding fragments of antibodies (e.g., single-chain antibodies, Fab and sFab fragments, F(ab')2, Fd fragments, Fv fragments, scFv, and domain antibody (dAb) fragments (see, for example, the whole being incorporated herein by reference, Wildt et al., Eur J.Immunol., 1996;26(3):629-39)), as well as complete antibodies. Antibodies may have the structural features of IgA, IgG, IgE, IgD, IgM (and their subtypes and combinations). Antibodies may originate from any source, including mice, rabbits, pigs, rats, and primates (human and non-human primates), as well as primate-like antibodies. Antibodies also include minibodies, humanized antibodies, chimeric antibodies, and the like.

[0070] As used herein, “antibody variable domain” refers to the light and heavy chain portions of an antibody molecule, including the amino acid sequences of the complementarity-determining regions (CDRs, i.e., CDR1, CDR2, and CDR3), as well as the framework region (FR). VH refers to the variable domain of the heavy chain. VL refers to the variable domain of the light chain. According to the methods used in this disclosure, the amino acid positions assigned to the CDRs and FRs may be defined according to Kabat (Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, Md., 1987 and 1991)) or according to IMGT nomenclature.

[0071] As used herein, the term “antibody binding site” refers to a polypeptide or domain comprising one or more CDRs of an antibody that can bind to an antigen. For example, the polypeptide comprises CDR3 (e.g., HCDR3). For example, the polypeptide comprises CDR1 and 2 (e.g., HCDR1 and 2) or CDR1-3 of the variable domains (e.g., HCDR1-3) of the antibody. In one example, the antibody binding site is provided by a single variable domain (e.g., a VH domain or a VL domain). In another example, the binding site comprises a VH / VL pair, or two or more such pairs.

[0072] As used herein, “OX40L antagonist antibody” or “OX40L antagonist antibody” refers to an antibody or its antigen-binding fragment that can inhibit and / or neutralize the biological signaling activity of OX40L, for example, by blocking or substantially reducing the binding of OX40 to OX40L.

[0073] As used herein, “buffer solution” refers to a chemical that can absorb a certain amount of acid or base without being subject to strong pH fluctuations.

[0074] As used herein, the term “cell” means in vitro, ex vivo, or in vivo cells. In some embodiments, ex vivo cells may be part of a tissue sample excised from an organism such as a mammal. In some embodiments, in vitro cells may be cells in a cell culture. In some embodiments, in vivo cells are cells residing in an organism such as a mammal.

[0075] As used herein, the term “dose” refers to a specific amount or quantity of a drug to be taken or recommended to be taken at a particular time. As used herein, it is typically expressed in mg of an antibody or fragment thereof. Alternatively, it may be expressed in terms of mg / kg, taking into account the patient’s body weight. “Daily dose” refers to the total amount of medication administered to an individual in a 24-hour period only.

[0076] As used herein, the term “dosage” refers to administering a specific amount, number, and frequency of doses over a specific period of time. “Dosage” means duration. “Medication regimen” is a treatment plan for administering a drug over a period of time.

[0077] As used herein, the terms “improve,” “to improve,” or “improve,” or their grammatical variations, as used in relation to behavior, refer to the ability of an object, including human or non-human animals, to achieve a measurable increase in performance in relation to a task used to test those behaviors.

[0078] As used herein, “injection” refers to the means of administration, including, for example, intravenous (IV) and subcutaneous injections. IV injections may be referred to as infusions. It is also used herein to refer to administration examples, where the administration is by injection, for example, in the phrase “one or more induction phase injections.” Each injection involves the administration of a certain dose of antibody or fragment thereof.

[0079] As used herein, “injection device” means a device designed to perform an injection, and an injection includes the step of temporarily fluidizing the injection device to human tissue, typically subcutaneous tissue. An injection further includes administering a certain amount of aqueous or liquid drug to the tissue and detaching or removing the injection device from the tissue. In some embodiments, the injection device may be an intravenous or IV device, which is a type of injection device used when the target tissue is blood in the circulatory system, for example, blood in a vein. Common but non-limiting examples of injection devices are needles and syringes.

[0080] As used herein, “instructions” means any written, printed, or graphic indication of an item on its immediate container, such as written material displayed on a vial containing a pharmaceutically active drug, or details of the use of a product contained in a formulation or kit containing the formulation of interest. Instructions describe a method of administration or treatment intended to be carried out.

[0081] As used herein, the terms “isolated antibody” or “purified antibody” refer to an antibody having one to four of the following characteristics, depending on its origin or source of origin: (1) not associated with any naturally associated components that accompany it in its natural state; (2) not containing, or substantially not containing, other proteins of the same species; (3) expressed by cells of a different species; or (4) not present in nature. An isolated antibody is substantially free from other antibodies with different antigen specificities.

[0082] As used herein, the term “preparation” in relation to antibodies means an antibody combined with a pharmaceutically acceptable excipient comprising at least one buffer, at least one stabilizer, at least one surfactant, and at least one chelating agent, with the pH as defined. As used herein, the term “preparation” may be used interchangeably with the term “composition.”

[0083] As used herein, “monoclonal antibody” refers to an antibody obtained from a substantially homogeneous population of antibodies, i.e., the individual antibodies within the population are identical except for naturally occurring mutations that may be present in small amounts. Monoclonal antibodies are highly specific and directed to a single antigenic site. Furthermore, in contrast to polyclonal antibody preparations, which typically contain different antibodies directed to different determinants (epitopes), each monoclonal antibody is directed to a single determinant on the antigen. The modifier “monoclonal” indicates the characteristic of an antibody obtained from a substantially homogeneous population of antibodies and should not be interpreted as requiring antibody production by any particular method. For example, monoclonal antibodies used in accordance with this disclosure may be produced by the hybridoma method first described by Kohler and Milstein, 1975, Nature 256:495, or by the recombinant DNA method as described in U.S. Patent No. 4,816,567. Monoclonal antibodies may also be isolated from phage libraries produced using techniques such as those described, for example, McCafferty et al., 1990, Nature 348:552-554. As used herein, “humanized” antibody refers to a form of non-human (e.g., mouse) antibody that is a chimeric immunoglobulin, immunoglobulin chain, or fragment thereof (e.g., Fv, Fab, Fab', F(ab')2, or other antigen-binding subsequences of the antibody) containing the smallest sequence derived from non-human immunoglobulin. In certain exemplary embodiments, the humanized antibody is a human immunoglobulin (recipient antibody) in which residues from the recipient's CDR are replaced with residues from the CDR of a non-human species (donor antibody), such as mouse, rat, or rabbit, having the desired specificity, affinity, and capabilities. The humanized antibody may also contain residues not found in the recipient antibody or the transferred CDR or framework sequence, but included to further refine and optimize antibody performance.

[0084] As used herein, the terms “level” and “levels” may be used synonymously with the terms “concentration” and “concentration.”

[0085] As used herein, the terms “patient,” “subject,” “animal,” or “host” refer to mammals. A subject may be a human, but may also be a mammal requiring veterinary treatment, such as livestock (e.g., dogs, cats, etc.), farm animals (e.g., cattle, sheep, poultry, pigs, horses, etc.), and laboratory animals (e.g., rats, mice, guinea pigs, etc.).

[0086] As used herein, the terms “peptide” and “polypeptide” are interchangeable herein and refer to compounds consisting of approximately 2 to approximately 90 amino acid residues (inclusive), where the amino group of one amino acid is linked to the carboxyl group of another amino acid by a peptide bond. Peptides can be derived or removed from native proteins, for example, by enzymatic or chemical cleavage, or prepared using conventional peptide synthesis techniques (e.g., solid-phase synthesis) or molecular biology techniques (see Sambrook et al., MOLECULAR CLONING:LAB.MANUAL (Cold Spring Harbor Press, Cold Spring Harbor, NY, 1989)). The "peptides" may include any suitable L-amino acids and / or D-amino acids, e.g., common α-amino acids (e.g., alanine, glycine, valine), non-α-amino acids (e.g., P-alanine, 4-aminobutyric acid, 6-aminocaproic acid, sarcosine, statins), and unusual amino acids (e.g., citrulline, homocitrulline, homoserine, norleucine, norvaline, ornithine). The amino, carboxyl, and / or other functional groups on the peptide may be free (e.g., unmodified) or protected with suitable protecting groups. Suitable protecting groups for amino and carboxyl groups, as well as means for adding or removing protecting groups, are known in the art. See, for example, Green and Wuts, PROTECTING GROUPS IN ORGANIC SYNTHESIS (John Wiley and Sons, 1991). The functional groups of the peptide may also be derivatized (e.g., alkylated) using methods known in the art.

[0087] As used herein, the terms “pharmaceutical preparation” or “drug preparation” refer to a preparation that is in a form that enables the biological activity of an active ingredient to be effective. The terms “pharmaceutical preparation” and “drug preparation” refer to a mixture or structure in which different chemical substances, including an active drug, are combined to form an unconventional product such as a sterile product, solution, powder, emulsion, capsule, tablet, granule, topical preparation, semi-solid or sustained-release preparation, or liquid, which forms the final pharmaceutical product. Pharmaceutical preparations are prepared according to a specific procedure, i.e., “process.” The drug formed will differ depending on the route of administration. As used herein, the term “preparation” in relation to antibodies may refer to an antibody combined with a pharmaceutically acceptable excipient, for example, containing at least one buffer, at least one stabilizer, at least one surfactant, and at least one chelating agent, with the pH as defined. Alternatively, the term “preparation” in relation to antibodies may refer to an antibody combined with a pharmaceutically acceptable excipient, for example, containing at least one buffer, at least one stabilizer, and at least one surfactant, with the pH as defined.

[0088] As used herein, the term “pharmaceutical preparation” is interchangeable with the term “pharmaceutical composition,” which further refers to an activator in combination with a pharmaceutically acceptable carrier, e.g., a carrier commonly used in the pharmaceutical industry. The phrase “pharmaceutically acceptable” is used herein to mean a compound, material, composition, or preparation and / or dosage form that, within the bounds of sound medical judgment, is suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic response, or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0089] As used herein, the term “pharmaceutically acceptable excipient” (vehicle, additive) means that which can be safely administered to a subject to provide an effective dose of the active ingredient being used. As used herein, the term “excipient” or “carrier” refers to an inert substance commonly used as a diluent, vehicle, preservative, binder, or stabilizer of a drug. As used herein, the term “diluent” refers to a pharmaceutically acceptable (safe for administration to humans and non-toxic) solvent that is useful in the preparation of aqueous formulations as used herein. Example diluents include, but are not limited to, sterile water and bacteriostatic water for injection (BWFI).

[0090] As used herein, the term “pharmaceutically acceptable carrier” means a pharmaceutically acceptable material, composition, formulation, or carrier, such as a liquid or solid filler, stabilizer, dispersant, suspension, diluent, excipient, thickener, solvent, or encapsulating material, which is involved in carrying or transporting a compound useful in this disclosure into or to a subject in order to perform its intended function. Typically, such a construct is carried or transported from one organ or part of the body to another organ or part of the body. Each carrier must be “acceptable” in the sense that it is compatible with the other components of the formulation, including the compound useful in this disclosure, and is not harmful to the subject. Some examples of materials that can serve as pharmaceutically acceptable carriers include sugars, e.g., lactose, glucose, and sucrose; starches, e.g., corn starch and potato starch; cellulose and its derivatives, e.g., sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; tragacanth powder; malt; gelatin; talc; excipients, e.g., cocoa butter and suppository wax; oils, e.g., peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols, e.g., propylene glycol; polyols, e.g., glycerin, sorbitol, mannitol, and polyethylene glycol; esters, e.g., ethyl oleate and ethyl laurate; agar; buffers, e.g., magnesium hydroxide and aluminum hydroxide; surfactants, alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol; phosphate buffer solution; and other non-toxic, suitable substances used in pharmaceutical formulations.

[0091] As used herein, the term “pharmaceutically acceptable carrier” includes all coatings, antimicrobial and antifungal agents, as well as absorption retarders, etc., that are physiologically acceptable to the subject, and whose activity and compatibility with the compounds useful in this disclosure. In certain circumstances, auxiliary active compounds may also be incorporated into the pharmaceutical formulation. A “pharmaceutically acceptable carrier” may further include pharmaceutically acceptable salts of the compounds disclosed herein. Other additional components that may be included in the pharmaceutical formulation are known in the art and are described, for example, in Remington's Pharmaceutical Sciences (Genaro, Ed., Mack Publishing Co., 1985, Easton, PA), which are incorporated herein by reference.

[0092] As used herein, the term “pharmaceutically acceptable salt” refers to a derivative of the disclosed compound, the parent compound modified by converting an existing acidic or base moiety to its salt form. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines, and alkali or organic salts of acidic residues such as carboxylic acids. Examples of pharmaceutically acceptable salts in this disclosure include conventional non-toxic salts of parent compounds formed from non-toxic inorganic or organic acids. Pharmaceutically acceptable salts in this disclosure can be synthesized from parent compounds containing basic or acidic moieties by conventional chemical methods. Generally, such salts can be prepared by reacting the free acidic or basic form of these compounds with a stoichiometric amount of a suitable base or acid in water, an organic solvent, or a mixture thereof, generally using non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile. The phrase “pharmaceutically acceptable salt” is not limited to a single or 1:1 salt. For example, “pharmaceutically acceptable salts” also include bis salts, e.g., bis hydrochloride. A list of suitable salts can be found in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418, and Journal of Pharmaceutical Science, 66, 2 (1977), each of which is incorporated herein by reference in whole.

[0093] As used herein, when referring to the polypeptides of this disclosure, the terms “part,” “fragment,” “variant,” “derivative,” and “analog” include any polypeptide that retains at least some biological activity (e.g., antigen binding) as referred herein.

[0094] As used herein, the terms “prevent” or “prevention” mean, if nothing has occurred, the absence of the onset of a disability or disease, or, if the onset of a disability or disease already exists, the absence of further onset of a disability or disease. The ability to prevent some or all of the symptoms associated with a disability or disease is also considered.

[0095] As used herein, the terms “to treat,” “to treat,” “to treat,” or “to improve” refer to a therapeutic action whose purpose is to reverse, reduce, improve, prevent, delay, or halt the progression or severity of a condition associated with a disease or disorder. The term “to treat” includes reducing or mitigating at least one adverse effect or symptom of a condition, disease, or disorder. Treatment is generally “effective” if one or more symptoms or clinical markers are reduced. Alternatively, treatment is “effective” if the progression of the disease is reduced or halted. That is, “treatment” includes not only improvement of symptoms or markers but also halt or at least delay in the progression or worsening of symptoms compared to what would be expected in the absence of treatment. Beneficial or desired clinical outcomes include, but are not limited to, relief of one or more symptoms, whether detectable or undetectable; reduction of the severity of the disease; stable (i.e., non-worsening) state of the disease; delay or slowing of disease progression; improvement or mitigation of the condition; remission (partial or complete); and / or reduction in mortality. The term "treatment" of a disease also includes providing relief from the symptoms or side effects of the disease (including palliative care). For a treatment to be effective, a complete cure is not intended. A method may, in certain aspects, also include a cure.

[0096] As used herein, the term "mg / kg" refers to the dose of a substance administered to an individual in milligrams per kilogram of body weight.

[0097] As used herein, “packaging” means how the components are organized and / or constrained into units suitable for distribution and / or use. Packaging may include, for example, boxes, bags, syringes, ampoules, vials, tubes, clamshell packaging, containers for maintaining barriers and / or sterility, labeling, etc.

[0098] As used herein, the term “recombinant human antibody” is intended to include all antibodies prepared, expressed, produced, or isolated by recombinant means, such as antibodies expressed using a recombinant expression vector transfected into host cells, antibodies isolated from a recombinant combinatorial human antibody library, antibodies isolated from an animal (e.g., mouse) that is transgenic for a human immunoglobulin gene, or prepared antibodies, such recombinant human antibodies may be subjected to in vitro mutagenesis.

[0099] As used herein, “compound” or “sterile compound” involves preparing a pharmaceutical product in an environment free from bacteria, viruses, or any other potentially infectious microorganisms. Sterile compound is used in preparations administered IV, by injection, or directly into the eye.

[0100] As used herein, the terms “systemic administration,” “administered systemically,” “peripheral administration,” and “administered peripherally” mean, as used herein, the administration of a compound, drug, or other substance other than direct administration to a target tissue (e.g., the nervous system) as a result of it entering the animal system and thus undergoing metabolism and other similar processes, such as subcutaneous administration.

[0101] As used herein, the terms “sequence identity,” “identity percentage,” “homology percentage,” or, for example, “sequences that are 80% identical,” refer to the degree to which sequences are identical nucleotide-wise or amino acid-wise across a comparison window. Thus, “sequence identity percentage” can be calculated by comparing two optimally aligned sequences across a comparison window, determining the number of positions in both sequences where identical nucleic acid bases (e.g., A, T, C, G, I) or identical amino acid residues (e.g., Ala, Pro, Ser, Thr, Gly, Val, Leu, Ile, Phe, Tyr, Trp, Lys, Arg, His, Asp, Glu, Asn, Gln, Cys, and Met) occur, calculating the number of matching positions, dividing the number of matching positions by the total number of positions in the comparison window (i.e., the window size), and multiplying the result by 100 to calculate the sequence identity percentage.

[0102] The calculation of sequence similarity or sequence identity between sequences (these terms are used interchangeably herein) can be performed as follows: To determine the percentage of identity between two amino acid sequences or two nucleic acid sequences, the sequences may be aligned for optimal comparison purposes (for example, gaps may be introduced in one or both of the first and second amino acid or nucleic acid sequences for optimal alignment, and non-homologous sequences may be ignored for comparison purposes). In a particular embodiment, the length of the reference sequence aligned for comparison purposes is at least 30% of the length of the reference sequence, e.g., at least 40%, at least 50%, at least 60%, or at least 70%, at least 80%, at least 90%, or 100%. Then, amino acid residues or nucleotides are compared at the corresponding amino acid or nucleotide positions. If a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, the molecules are identical at that position.

[0103] The percentage of identity between two arrays is a function of the number of identical positions shared by the arrays, taking into account the number of gaps that need to be introduced for optimal alignment of the two arrays and the length of each gap.

[0104] The comparison of sequences and the determination of the percentage of identity between two sequences can be achieved using mathematical algorithms. In some embodiments, the percentage of identity between two amino acid sequences is determined using the algorithm of Needleman and Wunsch (1970, J. Mol. Biol. 48:444-453), which is incorporated into the GAP program in the GCG software package, using either the Blossum 62 matrix or the PAM250 matrix, as well as gap weights of 16, 14, 12, 10, 8, 6, or 4, and length weights of 1, 2, 3, 4, 5, or 6. In another preferred embodiment, the percentage of identity between two nucleotide sequences is determined using the GAP program in the GCG software package, using the NWSgapdna.CMP matrix, as well as gap weights of 40, 50, 60, 70, or 80, and length weights of 1, 2, 3, 4, 5, or 6. Another exemplary set of parameters includes a Blossum 62 scoring matrix with a 12-gap penalty, a 4-gap length penalty, and a 5-frameshift gap penalty. The percentage of identity between two amino acid or nucleotide sequences can also be determined using the PAM120 weight residue table, a 12-gap length penalty, and a 4-gap penalty, using the algorithm of E. Meyers and W. Miller (1989, Cabios, 4:11-17), which is incorporated into the ALIGN program (version 2.0).

[0105] For example, the peptide sequences described herein can be used as "query sequences" to search public databases to identify, for example, other family members or related sequences. Such searches can be performed using the NBLAST and XBLAST programs (version 2.0) of Altschul, et al. (1990, J. Mol. Biol. 215:403-10). To obtain nucleotide sequences homologous to the nucleic acid molecules of this disclosure, a BLAST nucleotide search can be performed using the NBLAST program, score=100, and word length=12. A BLAST protein search can be performed using the XBLAST program, score=50, and word length=3 to obtain amino acid sequences homologous to the protein molecules of this disclosure. To obtain gapped alignment for comparison purposes, Gapped BLAST as described in Altschul et al. (Nucleic Acids Res. 25:3389-3402, 1997) can be used. When using BLAST and Gapped BLAST programs, you can use the initial configuration parameters for each program (e.g., XBLAST and NBLAST).

[0106] Definitions of common terms in cell biology and molecular biology can be found in “The Merck Manual of Diagnosis and Therapy”, 19th Edition (published by Merck Research Laboratories), 2006 (ISBN 0-911910-19-0), Robert S. Porter et al. (eds.), The Encyclopedia of Molecular Biology (published by Blackwell Science Ltd.), 1994 (ISBN 0-632-02182-9), Benjamin Lewin, Genes X (published by Jones & Bartlett Publishing), 2009 (ISBN-10:0763766321), Kendrew et al. (eds.), Molecular Biology and Biotechnology: a Comprehensive Desk Reference (published by VCH Publishers, Inc.), 1995 (ISBN 1-56081-569-8), and Current Protocols in Protein Sciences. This can be found in 2009, Wiley Intersciences, Coligan et al., eds.

[0107] Unless otherwise specified, standard procedures refer to, for example, Sambrook et al., Molecular Cloning: A Laboratory Manual (4th ed.), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA (2012), Davis et al., Basic Methods in Molecular Biology, Elsevier Science Publishing, Inc., New York, USA (1995), or Methods in Enzymology: Guide to Molecular Cloning Techniques Vol. 152, SLBerger and ARKimmel Eds., Academic Press Inc., San Diego, USA (1987), Current Protocols in Protein Science (CPPS) (John E. Coligan, et al., ed., John Wiley and Sons, Inc.), Current Protocols in Cell Biology (CPCB) (Juan S. Bonifacino et al. ed., John Wiley and Sons, Inc.), and Culture of Animal Cells: A Manual of Basic Technique. These methods are used as described in *Animal Cell Culture Methods* (Methods in Cell Biology, Vol. 57, Jennie P. Mather and David Barnes editors, Academic Press, 1st edition, 1998), and all of these are incorporated herein by reference in their entirety.

[0108] Other terms are defined herein in the descriptions of various embodiments of this disclosure.

[0109] Where any aspect or embodiment of this disclosure is described in terms of Markush groups or other alternative groupings, this disclosure includes not only the entire list of groups as a whole, but also each member of each individual group, all possible subgroups of a principal group, and principal groups in which one or more of the members of the group are absent. This disclosure also assumes the express exclusion of one or more of the members of the groups in the claims set forth below.

[0110] Pharmaceutical preparations As used herein, the term “pharmaceutical preparation” means a combination of at least one active ingredient (e.g., a small molecule, polymer, compound, etc.) capable of exerting a biological effect in humans or non-human animals, and at least one inactive ingredient that, when combined with the active ingredient or one or more additional inactive ingredients, is suitable for therapeutic administration to humans or non-human animals. As used herein, the term “preparation” means “pharmaceutical preparation” unless otherwise specifically indicated. A “pharmaceutically acceptable” excipient (vehicle, additive) is one that can be reasonably administered to the target mammal in order to provide an effective dose of the active ingredient used.

[0111] In various embodiments, the pharmaceutical formulations according to the present disclosure are in aqueous form comprising an antibody or antigen-binding fragment thereof that specifically binds to OX40L. More specifically, the present disclosure provides a pharmaceutical formulation comprising (a) a monoclonal antibody or antigen-binding fragment thereof that specifically binds to an OX40 ligand (OX40L mAb), (b) a stabilizer, (c) a surfactant, and (d) a buffer comprising histidine or a histidine salt. The pH of the aqueous pharmaceutical formulation may be in the range of about 5.5 to about 6.3, for example, about 5.8 to about 6.2, or the pH is about 6.0.

[0112] The antibody pharmaceutical formulations provided herein can be prepared by mixing an antibody of desired purity with one or more optionally selected pharmaceutically acceptable carriers or excipients (Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980)). In exemplary embodiments, the pharmaceutical formulation is provided in the form of an aqueous solution.

[0113] Aqueous pharmaceutical formulations have been shown to stably support antibodies or their antigen-binding fragments at high concentrations of approximately 125 mg / mL. The disclosed aqueous pharmaceutical formulations are suitable for administration to mammalian subjects by parenteral administration, including subcutaneous, intravenous, intramuscular, intraperitoneal, or intradermal injection.

[0114] Specific exemplary components and formulations included in this disclosure are described in detail below.

[0115] Antibodies or fragments thereof Depending on the various embodiments, antibodies or fragments contained in the formulations disclosed herein may include, but are not limited to, synthetic antibodies, monoclonal antibodies, recombinant antibodies, multispecific antibodies (including bispecific antibodies), human antibodies, humanized antibodies, chimeric antibodies, intrabodies, single-chain Fv(scFv) (e.g., monospecific, bispecific, etc.), camelid antibodies, Fab fragments, F(ab') fragments, disulfide-linked Fv(sdFv), anti-idiotype (anti-Id) antibodies, and any of the above epitope-linked fragments that can bind to OX40L (OX40L) or OX40.

[0116] The term "specifically binds" or similar means that the antibody or its antigen-binding fragment forms a complex with an antigen such as hOX40L, which is relatively stable under physiological conditions. Specific binding is at least about 1 × 10⁻¹⁶. -6It can be characterized by a dissociation constant of M or greater. Methods for determining whether two molecules specifically bind are well known in the art and include, for example, equilibrium dialysis and surface plasmon resonance. However, isolated antibodies that specifically bind to hOX40L may have cross-reactivity to other antigens, such as OX40L molecules from other species (orthologs). In the context of the present invention, multispecific (e.g., bispecific) antibodies that bind to human OX40L, as well as one or more additional antigens, are considered to "specifically bind" to human OX40L. Furthermore, isolated antibodies may substantially contain no other cellular material or chemicals.

[0117] In particular, antibodies included in the formulations disclosed herein include immunoglobulin molecules and molecules containing immunologically active portions of immunoglobulin molecules, i.e., antigen-binding sites that specifically bind to the hOX40L antigen. The immunoglobulins provided herein may be any type (e.g., IgG, IgE, IgM, IgD, IgA, and IgY), class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), or subclass of immunoglobulin molecules. In certain embodiments, the antibody according to this disclosure is an IgG antibody, e.g., IgG1 or IgG4. In certain embodiments, the antibody according to this disclosure includes a human gamma 4 constant region. In one embodiment, the antibody is human IgG4, and its heavy chain constant region IgG4-PE includes Leu235Glu and Ser228Pro Fc mutations.

[0118] Antibody variants and derivatives include antibody fragments that retain the ability to specifically bind to an epitope. Exemplary fragments include: Fab fragments; Fab' (an antibody fragment containing a single anti-binding domain and an additional portion of the heavy chain passing through a hinge region); F(ab')2 (two Fab' molecules linked by an interchain disulfide bond within the hinge region of the heavy chain; the Fab' molecules can be directed to the same or different epitopes); bispecific Fab (a Fab molecule having two antigen-binding domains, each directed to a different epitope); single-stranded Fab chains containing a variable region, also known as scFv; disulfide-bonded Fv; or dsFv; camelid VH (a variable, antigen-binding variable single heavy chain of an antibody, where several amino acids at the VH interface are found in the naturally occurring heavy chain of camel antibodies); Examples include: specific regions); bispecific scFv (an scFv or dsFv molecule having two antigen-binding domains, each of which may be directed to a different epitope); diabody (a dimerized scFv formed when the VH domain of a first scFv is assembled with the VL domain of a second scFv, and the VL domain of the first scFv is assembled with the VH domain of the second scFv; the two antigen-binding regions of the diabody may be directed to the same or different epitopes); and tribody (a trimerized scFv formed in a similar manner to the diabody, but with three antigen-binding domains made up in a single complex; the three antigen-binding domains may be directed to the same or different epitopes). Derivatives of antibodies also include one or more CDR sequences of the antibody-binding site. If two or more CDR sequences are present, they may be linked together on the scaffold. In certain embodiments, the antibody includes a single-stranded Fv ("scFv"). scFv is an antibody fragment containing the VH and VL domains of an antibody, where these domains are present within a single polypeptide chain. Generally, scFv polypeptides further contain a polypeptide linker between the VH and VL domains, which allows the scFv to form a desired structure for antigen binding.For an overview of scFv, please refer to Pluckthun in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds. Springer-Verlag, New York, pp. 269-315 (1994).

[0119] In some embodiments, the antibodies contained in the formulations disclosed herein may be of any animal origin, including birds and mammals (e.g., humans, mice, donkeys, sheep, rabbits, goats, guinea pigs, camels, horses, or chickens). In certain embodiments, the antibodies may be human or humanized monoclonal antibodies. As used herein, “human” antibodies include antibodies having the amino acid sequence of human immunoglobulins and include antibodies isolated from human immunoglobulin libraries or from mice expressing antibodies from human genes.

[0120] In some embodiments, the antibody is a fully human antibody, such as a fully human antibody that specifically binds to the hOX40L polypeptide, hOX40L polypeptide fragment, or hOX40L epitope. Such fully human antibodies are advantageous over fully human mouse antibodies (or other fully or partially non-human species antibodies), humanized antibodies, or chimeric antibodies, as they minimize the occurrence of undesirable or unwanted side effects, such as an immune response directed towards a non-fully human antibody (e.g., an anti-hOX40L antibody derived from another species) when administered to a subject.

[0121] In some embodiments, the antibody may be monospecific, bispecific, triplicate, or more highly multispecific. A multispecific antibody may be specific to different epitopes of the hOX40L polypeptide, or it may be specific to both the hOX40L polypeptide and a heterologous epitope, such as a heterologous polypeptide or a solid support material. In an exemplary embodiment, the antibody provided herein is monospecific to a given epitope of the hOX40L polypeptide and does not specifically bind to other epitopes.

[0122] In certain embodiments, the isolated antibody provided herein specifically binds to the hOX40L epitope, and the binding of the antibody to the hOX40L epitope is competitively blocked (e.g., dose-dependently) by the antibody or fragment of the disclosure. The antibody may or may not be a whole human antibody. In exemplary embodiments, the antibody is a whole human monoclonal anti-hOX40L antibody. In particularly exemplary embodiments, the antibody is a whole human, monoclonal, antagonist anti-hOX40L antibody. Exemplary competitive blockage tests that can be used are provided in the examples herein.

[0123] In some embodiments, the antibody or fragment of the Disclosure competes with the OX40 receptor (or its fusion protein) for binding to cell surface-expressed hOX40L (e.g., in a dose-dependent manner). In other embodiments, the antibody or fragment of the Disclosure competes with the OX40 receptor (or its fusion protein) for binding to soluble hOX40L (e.g., in a dose-dependent manner). In one embodiment, the antibody or fragment partially or completely inhibits the binding of hOX40 to cell surface-expressed OX40L, such as hOX40L. In another embodiment, the antibody partially or completely inhibits the binding of hOX40 to soluble hOX40L.

[0124] In some embodiments, the antibody or antigen-binding fragment of the antibody is a fully human monoclonal antibody that specifically binds to hOX40L, for example, a fully human monoclonal antagonist antibody.

[0125] In some embodiments, an antibody or fragment binds to an hOX40L epitope, which is a three-dimensional surface feature of the hOX40L polypeptide (e.g., the trimer form of the hOX40L polypeptide). The region of the hOX40L polypeptide contributing to the epitope may be a sequence of amino acids of the polypeptide, or the epitope may be formed from two or more discontinuous regions of the polypeptide. The hOX40L epitope may exist in (a) the trimer form of hOX40L ("trimeric hOX40L epitope"), (b) the monomer form of hOX40L ("Ca monomer hOX40L epitope"), (c) both the trimer and monomer forms of hOX40L, (d) the trimer form but not the monomer form of hOX40L, or (e) the monomer form but not the trimer form of hOX40L.

[0126] For example, in some embodiments, the epitope exists or is available for binding only in its trimer (natural) form, but does not exist or is not available for binding in its monomeric (denatured) form by the anti-hOX40L antibody. In other embodiments, the hOX40L epitope is a linear feature of the hOX40L polypeptide (e.g., the trimer or monomeric form of the hOX40L polypeptide). The antibodies provided herein can specifically bind to (a) the monomeric form of hOX40L epitope, (b) the trimer form of hOX40L epitope, (c) the monomeric form of hOX40L but not the trimer form, (d) the trimer form of hOX40L but not the monomer form, or (e) both the monomeric and trimer forms of hOX40L. In exemplary embodiments, the antibodies provided herein specifically bind to the trimer epitope of hOX40L but not to the monomer epitope of hOX40L.

[0127] In some embodiments, the antibody specifically binds to the hOX40L epitope, and the antibody comprises derivatives of the VH domain, VH CDR, VL domain, and VL CDR described herein that specifically bind to the hOX40L antigen. Some embodiments also provide antibodies comprising derivatives of antibodies that specifically bind to the hOX40L epitope. Using standard techniques known to those skilled in the art, mutations can be introduced into the nucleotide sequence encoding the molecule, including, for example, site-directed mutagenesis and PCR-mediated mutagenesis resulting in amino acid substitutions. In certain exemplary embodiments, the derivatives include fewer than 25 amino acid substitutions, fewer than 20 amino acid substitutions, fewer than 15 amino acid substitutions, fewer than 10 amino acid substitutions, fewer than 5 amino acid substitutions, fewer than 4 amino acid substitutions, fewer than 3 amino acid substitutions, or fewer than 2 amino acid substitutions compared to the original molecule. In another embodiment, the derivative has a conservative amino acid substitution. In certain exemplary embodiments, the derivative has a conservative amino acid substitution that occurs at one or more predicted non-essential amino acid residues. Alternatively, mutations can be introduced randomly along all or part of the coding sequence, for example, by saturation mutagenesis. The resulting mutants can then be screened for biological activity to identify those that retain activity. After mutagenesis, the encoded protein can be expressed, and its activity can be determined.

[0128] In another embodiment, the antibody that specifically binds to the hOX40L epitope comprises a variable domain amino acid sequence that is at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the variable domain amino acid sequence in the sequence listing.

[0129] In certain embodiments, the antibody is a fully human anti-human antibody, such as a fully human monoclonal antibody. Fully human antibodies can be produced by any method known in the art. Exemplary methods include immunization of a transgenic animal (e.g., mouse) with the hOX40L antigen (any hOX40L polypeptide capable of inducing an immune response, and optionally conjugated to a carrier) in the absence of endogenous immunoglobulin production, which can produce a repertoire of human antibodies. See, for example, Jakobovits et al., (1993) Proc. Natl. Acad. Sci., 90:2551, Jakobovits et al., (1993) Nature, 362:255 258 (1993), and Bruggermann et al., (1993) Year in Immunol., 7:33. Other methods for producing fully human anti-hOX40L antibodies can be found in the examples provided herein.

[0130] Alternatively, fully human antibodies may be generated through in vitro screening of phage display antibody libraries. See, for example, Hoogenboom et al., J.Mol.Biol., 227:381 (1991) and Marks et al., J.Mol.Biol., 222:581 (1991), incorporated herein by reference. Various antibody-containing phage display libraries are described and can be readily prepared by those skilled in the art. The libraries may contain a diversity of human antibody sequences, such as human Fab, Fv, and scFv fragments, which can be screened against appropriate targets.

[0131] This disclosure encompasses antibodies or fragments conjugated to therapeutic portions ("immune conjugates") such as cytotoxins, chemotherapeutic agents, immunosuppressants, or radioisotopes. Examples of cytotoxic agents include any agent harmful to cells. Examples of suitable cytotoxic and chemotherapeutic agents for forming immune complexes are known in the art, see, for example, WO05 / 103081, which is incorporated herein by reference in its entirety.

[0132] In some embodiments, antibodies and fragments may include antibodies and fragments that are chemically modified, i.e., modified by covalent bonding of any kind of molecule to the antibody. For example, but not limited to, antibody derivatives include antibodies that have been chemically modified by, for example, glycosylation, acetylation, pegylation, phosphorylation, amidation, derivatization with known protecting / blocking groups, proteolytic cleavage, binding to cell ligands or other proteins. Any of the many chemical modifications may be carried out by known techniques, including, but not limited to, specific chemical cleavage, acetylation, formulation, and metabolic synthesis of tunicamycin. Additionally, antibodies may contain one or more non-classical amino acids.

[0133] In some embodiments, antibodies that specifically bind to the hOX40L antigen include framework regions (e.g., human or non-human fragments) known to those skilled in the art. The framework regions may be, for example, naturally occurring or consensus framework regions. In certain embodiments, the framework region of the antibodies of this disclosure is human (for example, see Chothia et al., 1998, J.Mol.Biol.278:457-479, which is incorporated herein by reference in its entirety). See also Kabat et al. (1991) Sequences of Proteins of Immunological Interest (USD Department of Health and Human Services, Washington, DC) 5th ed.

[0134] In some embodiments, the antibody that specifically binds to the hOX40L antigen is one or more amino acid sequences of the CDR in the sequence listing (i.e., SEQ ID NO: 4, SEQ ID NO: 10, SEQ ID NO: 36, SEQ ID NO: 42, SEQ ID NO: 68, SEQ ID NO: 74, SEQ ID NO: 96, or SEQ ID NO: 102; in particular for HCDR1, SEQ ID NO: 36 or SEQ ID NO: 42; SEQ ID NO: 6, SEQ ID NO: 12, SEQ ID NO: 38, SEQ ID NO: 44, SEQ ID NO: 70, SEQ ID NO: 76, SEQ ID NO: 98, or SEQ ID NO: 104; in particular for HCDR2, SEQ ID NO: 38 or SEQ ID NO: 44; SEQ ID NO: 8, sequence Number 14, SEQ ID NO: 40, SEQ ID NO: 46, SEQ ID NO: 72, SEQ ID NO: 78, SEQ ID NO: 100, or SEQ ID NO: 106; especially for HCDR3, SEQ ID NO: 40 or SEQ ID NO: 46; SEQ ID NO: 18, SEQ ID NO: 24, SEQ ID NO: 50, SEQ ID NO: 56, SEQ ID NO: 82, SEQ ID NO: 88, SEQ ID NO: 110, or SEQ ID NO: 116; especially for LCDR1, SEQ ID NO: 50 or SEQ ID NO: 56; SEQ ID NO: 20, SEQ ID NO: 26, SEQ ID NO: 52, SEQ ID NO: 58, SEQ ID NO: 84, SEQ ID NO: 90, SEQ ID NO: 112, or SEQ ID NO: 118; especially for LCDR2, Sequence No. 52 or SEQ ID NO. 58; and SEQ ID NO. 22, SEQ ID NO. 28, SEQ ID NO. 54, SEQ ID NO. 60, SEQ ID NO. 86, SEQ ID NO. 92, SEQ ID NO. 114, or SEQ ID NO. 120, in particular SEQ ID NO. 54 or SEQ ID NO. 60 for LCDR3), and including human framework regions having one or more amino acid substitutions in one, two, three, or more of the following residues: (a) rare framework residues that differ between the mouse antibody framework (i.e., donor antibody framework) and the human antibody framework (i.e., acceptor antibody framework); (b) Vernier zone residues where they differ between the donor antibody framework and the acceptor antibody framework; (c) interchain packing residues at the VH / VL interface that differ between the donor antibody framework and the acceptor antibody framework; (d) canonical residues that differ between the donor antibody framework and the acceptor antibody framework sequence, in particular framework regions essential to defining the canonical class of the mouse antibody CDR loop; (e) residues adjacent to the CDR; (g) residues that can interact with the antigen;(h) Residues that can interact with the CDR; and (i) Contact residues between the VH domain and the VL domain. In a particular embodiment, an antibody that specifically binds to an hOX40L antigen containing a human framework region having one or more amino acid substitutions in one, two, three, or more of the residues identified above is an antagonistic hOX40L antibody.

[0135] In some embodiments, an antibody that specifically binds to the hOX40L antigen includes the amino acid sequence of the VH domain and / or VL domain in the sequence listing (i.e., for the VH domain, SEQ ID NO: 2, SEQ ID NO: 34, SEQ ID NO: 66, or SEQ ID NO: 94, particularly SEQ ID NO: 34; for the VL domain, SEQ ID NO: 16, SEQ ID NO: 48, SEQ ID NO: 80, or SEQ ID NO: 108, particularly SEQ ID NO: 48), but has mutations (e.g., one or more amino acid substitutions) within the framework region. In a particular embodiment, an antibody that specifically binds to the hOX40L antigen includes the amino acid sequence of the VH domain and / or VL domain or its antigen-binding fragment, having one or more amino acid residue substitutions in the framework region of the VH domain and / or VL domain.

[0136] In some embodiments, the antibody that specifically binds to the hOX40L antigen includes the heavy chain complementarity-determining region (HCDR) 1 of SEQ ID NO: 42, the HCDR2 of SEQ ID NO: 44, and the HCDR3 of SEQ ID NO: 46, as well as the light chain complementarity-determining region (LCDR) 1 of SEQ ID NO: 56, the LCDR2 of SEQ ID NO: 58, and the LCDR3 of SEQ ID NO: 60.

[0137] In some embodiments, an antibody that specifically binds to the hOX40L epitope comprises the amino acid sequence of the VH domain and / or VL domain in the sequence listing (i.e., for the VH domain, SEQ ID NO: 2, SEQ ID NO: 34, SEQ ID NO: 66, or SEQ ID NO: 94, particularly SEQ ID NO: 34; for the VL domain, SEQ ID NO: 16, SEQ ID NO: 48, SEQ ID NO: 80, or SEQ ID NO: 108, particularly SEQ ID NO: 48), but has mutations (e.g., one or more amino acid substitutions) within the framework region. In a particular embodiment, an antibody that specifically binds to the hOX40L antigen comprises the amino acid sequence of the VH domain and / or VL domain of the antibody disclosed in the Examples or its antigen-binding fragment, having one or more amino acid residue substitutions in the framework region of the VH domain and / or VL domain.

[0138] In another embodiment, an antibody that specifically binds to the hOX40L epitope comprises, for the VH domain, a variable domain amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to SEQ ID NO: 2, SEQ ID NO: 34, SEQ ID NO: 66, or SEQ ID NO: 94, in particular to SEQ ID NO: 34; and for the VL domain, a variable domain amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to SEQ ID NO: 48.

[0139] In one embodiment, an antibody or fragment thereof that specifically binds to the hOX40L antigen comprises an amino acid sequence of a VH domain and / or a VL domain, wherein the VH domain comprises the amino acid sequence described in SEQ ID NO: 34, and the VL domain comprises the amino acids described in SEQ ID NO: 48.

[0140] In one embodiment, an antibody or fragment thereof that specifically binds to the hOX40L antigen is a fully human antibody comprising a heavy chain and a light chain, wherein the heavy chain comprises a variable (VH) domain having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity to the amino acid sequence of SEQ ID NO: 34, and the light chain comprises a variable (VH) domain having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity to the amino acid sequence of SEQ ID NO: 48, as disclosed in US10,669,342 (WO2015 / 132580).

[0141] In one embodiment, the antibody is human IgG4, and its heavy chain constant region IgG4-PE contains Leu235Glu and Ser228Pro Fc mutations. In another embodiment, the heavy chain constant region is IgG4-PE.

[0142] In one embodiment, the antibody comprises a heavy chain having the amino acid sequence of SEQ ID NO: 62 and a light chain having the amino acid sequence of SEQ ID NO: 64. In one embodiment, the antibody is amriterimab or a variant thereof.

[0143] In some embodiments, a fusion protein comprising an antibody provided herein that specifically binds to the hOX40L antigen and a heterologous polypeptide may be used. In some embodiments, the heterologous polypeptide to which the antibody is fused is useful for targeting the antibody against cells having cell surface expression of hOX40L.

[0144] Optionally, an antibody or fragment may bind specifically to hOX40L with an affinity (apparent affinity, Kd) in the range of <1 mM, 1000 nM to 100 nM, 100 nM to 10 nM, 10 nM to 1 nM, 1000 pM to 500 pM, 500 pM to 200 pM, <200 pM, 200 pM to 150 pM, 200 pM to 100 pM, 100 pM to 10 pM, 10 pM to 1 pM, for example, 1 mM to 1 pM (e.g., 1 mM to 100 pM, 10 nM to 100 pM, 1 nM to 10 pM, or 100 pM to 1 pM), as determined by SPR (for example, under the SPR conditions disclosed herein).

[0145] In some embodiments, the antibodies in the pharmaceutical formulations and / or formulated antibodies provided herein comprise one or more charge variants (also called species or isoforms). Charge variants are typically described as primary charge variants (or species), acidic variants (or species), and basic variants (or species). The primary charge variant refers to the most common charge variant in a given antibody batch. Acidic variants have a lower isoelectric point (pI) compared to primary charge variants, and basic variants have a higher pH compared to primary charge variants. Charge variants may result from post-translational modifications to the amino acid sequences of HC and / or LC. For example, increased deamidation, glycation, and sialylation may cause a decrease in the pI of the antibody.

[0146] The charge variants of antibodies provided herein can be separated and quantified using methods known in the art for separating polypeptides based on charge. For example, in some embodiments, a weak cation exchange column can be used in a high-performance liquid chromatography (HPLC) system with a phosphate / sodium chloride gradient buffer. In this system, after loading the antibody onto the column, the acidic variant of the antibody elutes first, followed by the main charge variant, and then the basic variant of the antibody. In another embodiment, capillary isoelectric focusing (cIEF) can be used. Capillary isoelectric focusing is a method for separating proteins by their isoelectric point (pI) value. In cIEF, the antibody sample moves to its isoelectric point on the pH gradient in the capillary, thereby degrading different charge variants along the length of the capillary. To identify and characterize cIEF variants, the charge variants can be isolated by strong cation exchange chromatography (SCX) and analyzed by cIEF. Images of the degraded charge variants within the capillaries are acquired using a charge-coupled device camera with whole-column detection and absorbance measurement at 280 nm. The charge variant distribution of the test sample is compared to a reference standard using cIEF to identify the charge variant of the antibody. In some embodiments, cIEF is used as a quality control means to confirm the identity of the antibody during commercial production, and by determining the charge variant distribution of the antibody compared to a reference standard, e.g., a test antibody having a charge variant distribution pattern within a given distribution pattern, or a charge variant distribution pattern compared to the distribution pattern of the reference standard.

[0147] The amount of antibody or antigen-binding fragment contained in the pharmaceutical formulations of this disclosure may vary depending on the desired specific characteristics of the formulation, as well as the specific circumstances and purposes in which the formulation is intended to be used. In certain embodiments, the pharmaceutical formulation is an aqueous formulation containing antibody in a concentration ranging from about 1 mg / mL to about 150 mg / mL, including all partial ranges between them, such as about 20 mg / mL to about 140 mg / mL, for example, 28 ± 4.2 mg / mL to 138 mg / mL ± 20.7 mg / mL, or 31 ± 4.65 mg / mL to 125 mg / mL ± 18.75 mg / mL. In some embodiments, the antibody or antigen-binding fragment is present at concentrations of 28 ± 4.2 mg / mL, 31 mg / mL ± 4.65 mg / mL, 125 mg / mL ± 18.75 mg / mL, or 138 mg / mL ± 20.7 mg / mL. In one embodiment, the antibody or its antigen-binding fragment is present at concentrations of 28 mg / mL, 31 mg / mL, 125 mg / mL, or 138 mg / mL.

[0148] In a particular embodiment, the pharmaceutical formulation is approximately 75 mg / mL to approximately 138 mg / mL, for example, 75 (±10%) mg / mL to 138 (±10%) mg / mL, 85 (±10%) mg / mL to 138 (±10%) mg / mL, 95 (±10%) mg / mL to 138 (±10%) mg / mL, 105 (±10%) mg / mL to 138 (±10%) mg / mL, 115 (±10%) mg / mL to 138 (±10%) mg / mL, 125 This is an aqueous preparation containing antibodies at concentrations in the following ranges: (±10%) mg / mL to 138 (±10%) mg / mL, 75 (±10%) mg / mL to 128 (±10%) mg / mL, 75 (±10%) mg / mL to 118 (±10%) mg / mL, 75 (±10%) mg / mL to 108 (±10%) mg / mL, 75 (±10%) mg / mL to 98 (±10%) mg / mL, or 75 (±10%) mg / mL to 88 (±10%) mg / mL. For example, the formulations of this disclosure may include an antibody or antigen-binding fragment thereof that specifically binds to OX40L, such as hOX40L, in concentrations of 75 (±10%) mg / mL, 85 (±10%) mg / mL, 95 (±10%) mg / mL, 105 (±10%) mg / mL, 115 (±10%) mg / mL, 125 (±10%) mg / mL, or 135 (±10%) mg / mL.

[0149] In one embodiment, the formulation of the Disclosure comprises approximately 125 mg / mL, for example, 125 (±10%) mg / mL, of an antibody or antigen-binding fragment that specifically binds to hOX40L. In one embodiment, the formulation of the Disclosure comprises approximately 125 mg / mL of amliterimab or a variant thereof. In one embodiment, the formulation of the Disclosure comprises approximately 75 mg / mL of amliterimab or a variant thereof.

[0150] Excipients and pH In various embodiments, the pharmaceutical formulations of the Disclosure further comprise one or more excipients, such as stabilizers, surfactants, and buffers containing histidine or histidine salts, which are non-therapeutic agents added to the formulation to provide the formulation containing the anti-hOX40L antibody described herein with a desired consistency, viscosity, or stabilizing effect. In some embodiments, the pharmaceutical formulations provided herein comprise an antibody and a formulated antibody comprising one or more of the following excipients: sucrose, L-histidine, and polysorbate 80 (PS80).

[0151] The amount of each excipient may also vary depending on the desired specific properties of the formulation, as well as the specific circumstances and purposes in which the formulation is intended to be used. In some circumstances, the concentration of each excipient may be selected so that the pharmaceutical formulation can be diluted for administration by infusion.

[0152] surfactant In certain embodiments, the surfactant that may be included in the formulation is a nonionic surfactant selected from polysorbate and / or polxamer. In certain embodiments, the surfactant included in the pharmaceutical formulation is polysorbate 80 (PS80).

[0153] The amount of surfactant contained in the pharmaceutical formulations of this disclosure may vary depending on the desired specific properties of the formulation, as well as the specific circumstances and purposes for which the formulation is intended. In some embodiments, the surfactant is in an amount that stabilizes the antibody under rough handling or stirring conditions, such as vortexing. In some embodiments, “stabilizing” means preventing the formation of aggregated antibodies exceeding 3% (on a molar basis) of the total amount of antibody during the process of rough handling or stirring, such as vortexing. In some embodiments, the rough handling is vortexing a solution containing the antibody and an organic cosolvent for about 60 minutes or about 120 minutes.

[0154] In some embodiments, PS80 or PS20 is present in a pharmaceutical formulation at concentrations of approximately 0.005% (w / v) to approximately 0.1% (w / v), for example, 0.005% (w / v) ± 0.0015% to 0.1% ± 0.03% (w / v). For example, PS80 or PS20 may be present at concentrations of approximately 0.0085% (w / v), approximately 0.01% (w / v), approximately 0.02% (w / v), approximately 0.03% (w / v), approximately 0.04% (w / v), approximately 0.05% (w / v), approximately 0.06% (w / v), approximately 0.07% (w / v), approximately 0.08% (w / v), approximately 0.09% (w / v), or approximately 0.1% (w / v). In certain exemplary embodiments, PS80 is present in the formulation at a concentration of 0.04%(w / v) ± 0.006%(w / v). In certain exemplary embodiments, PS80 is present in the formulation at a concentration of 0.04%(w / v). In certain exemplary embodiments, PS80 is present in the formulation at a concentration of 0.06%(w / v) ± 0.009%(w / v). In certain exemplary embodiments, PS80 is present in the formulation at a concentration of 0.06%(w / v). In some embodiments, PS80 is present in the formulation at a concentration of 0.04%(w / v).

[0155] Stabilizer In some embodiments, the stabilizer in the pharmaceutical formulation of the Disclosure comprises mannitol and / or sucrose. In some embodiments, the stabilizer in the pharmaceutical formulation of the Disclosure is mannitol and / or sucrose. In certain embodiments, the stabilizer is sucrose. In some embodiments, sucrose is present in the pharmaceutical formulation at a concentration of about 200 mM to about 250 mM, for example, 220 mM ± 33 mM to 250 mM ± 37.5 mM. In some embodiments, the pharmaceutical formulation contains 220 mM sucrose.

[0156] In some embodiments, sucrose is present at a concentration that stabilizes the antibody under certain conditions. In some embodiments, such conditions may include thermal stress.

[0157] Buffer or buffer system The pharmaceutical formulations of this disclosure may also include buffers or buffer systems that help maintain a stable pH and assist in stabilizing antibodies. As used herein, the term “buffer” refers to a pharmaceutically acceptable buffer that maintains a stable pH or resists changes in the pH of the solution. In exemplary embodiments, the buffer contains histidine. In the context of this disclosure, “histidine buffer” or “histidine-containing buffer” refers to a buffer containing the amino acid histidine. Examples of histidine buffers include histidine chloride, histidine acetate, histidine phosphate, and histidine sulfate. In exemplary embodiments, the histidine buffer is prepared by dissolving L-histidine and L-histidine hydrochloride (e.g., monohydrate) in predetermined amounts and ratios. In one embodiment, the histidine buffer is prepared by titrating L-histidine (free base, solid) with dilution hydrochloric acid. The term “histidine” is used interchangeably with “histidine buffer” throughout this disclosure.

[0158] In some embodiments, L-histidine or histidine hydrochloride is present. In some embodiments, the concentration of L-histidine or histidine hydrochloride is about 10 mM, for example, 10 mM ± 3 mM. In some embodiments, the concentration of L-histidine or histidine hydrochloride is about 20 mM, for example, 20 mM ± 3 mM.

[0159] Chelating agents In some embodiments, the pharmaceutical formulations of the present disclosure may include one or more chelating agents, such as metal chelating agents. Metal chelating agents can protect the chemical stability of proteins under metal exposure.

[0160] In certain embodiments, the pharmaceutical formulation further comprises a chelating agent selected from the group consisting of ethylenediaminetetraacetic acid (EDTA), diethylenetrimenpentaacetic acid (DTPA), and their salts, and any combination thereof. In certain embodiments, the chelating agent is selected from EDTA and / or DTPA.

[0161] In a particular embodiment, the concentration of chelating agents such as EDTA and DTPA is approximately 10 μM, for example, 10 μM ± 1.5 μM.

[0162] Additional ingredients In some embodiments, the pharmaceutical formulations of the present disclosure may include one or more additional excipients.

[0163] However, in some embodiments, the pharmaceutical formulation does not require any additional excipients, such as arginine, which help maintain the reduced viscosity or decrease the viscosity of the formulation disclosed herein.

[0164] In some embodiments, the formulation does not contain sodium chloride.

[0165] In some embodiments, the pharmaceutical formulations according to this disclosure consist of, or are essentially composed of, the antibodies, sucrose, polysorbate 80, and L-histidine or histidine hydrochloride, as disclosed herein, but each component is present in the amounts described herein.

[0166] In some embodiments, the pharmaceutical formulations according to this disclosure consist of, or are essentially composed of, the antibodies, sucrose, polysorbate 80, and L-histidine or histidine hydrochloride disclosed herein, wherein each component is present in the amounts described herein.

[0167] pH The pH of a pharmaceutical formulation is optimized to maintain antibody stability, for example, when the pharmaceutical formulation is in aqueous form. In some embodiments, the pH of the formulation is about 5.5 to about 6.3, for example, 5.5 ± 0.2 to 6.3 ± 0.2. In some embodiments, the formulation has a pH of about 5.8 to about 6.2. For example, formulations of the present disclosure may have a pH of about 5.5, about 5.6, about 5.7, about 5.8, about 5.9, about 6.0, about 6.1, about 6.2, about 6.3, about 6.4, or about 6.5. In a particular exemplary embodiment, the pH of the formulation is about 6.0 ± 0.2. In one embodiment, the pH of the formulation is about 6.0 or 6.2.

[0168] Stability and other attributes The excipients and their amounts described herein have been demonstrated to provide desired consistency, viscosity, or stabilizing effects to pharmaceutical formulations containing anti-hOX40L antibody or its antigen-binding fragments at concentrations of approximately 75 mg / mL to approximately 138 mg / mL, for example, 75 mg / mL ± 11.25 mg / mL to 125 mg / mL ± 18.75 mg / mL or up to 138 mg / mL ± 20.7 mg / mL. Aqueous pharmaceutical formulations in various embodiments are typically particle-free or substantially particle-free.

[0169] In various embodiments, the pharmaceutical formulations of this disclosure typically exhibit a high level of stability. As used herein with respect to pharmaceutical formulations, the term “stable” means that the antibodies within the pharmaceutical formulation retain an acceptable degree of chemical structure or biological function after storage under defined conditions. A formulation may be stable even if the antibodies contained therein do not retain 100% of their chemical structure or biological function after storage for a defined amount of time. Under certain circumstances, retention of about 90%, 95%, 96%, 97%, 98%, or 99% of the structure or function of an antibody after storage for a specified time may be considered “stable.”

[0170] Stability can be measured, in particular, by determining the percentage of native antibodies remaining in the formulation after storage at a specified temperature for a specified time. The percentage of native antibodies can be determined, in particular, by size exclusion chromatography (e.g., size exclusion high-performance liquid chromatography [SE-HPLC]), where native proteins mean non-aggregation and non-degradation. "Acceptable stability," where the phrase is used herein, means that at least 85% of the native form of the antibody can be detected in the formulation after storage at a given temperature for a defined amount of time.

[0171] In certain embodiments, at least about 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the native form of the antibody can be detected in the formulation by size exclusion chromatography after storage for a defined time period. The defined time period after which stability is measured can be at least 14 days, at least 28 days, at least 1 month, at least 2 months, at least 3 months, at least 4 months, and a forecast of at least 1 year at about 5°C. The defined temperature at which the pharmaceutical formulation can be stored when assessing stability may be any temperature between about -80°C and about 40°C, for example, it can be stored at about -80°C, about -65°C, about -30°C, about -20°C, about 0°C, about 4°C to 8°C, about 5°C, about 25°C, about 35°C, about 37°C, or about 40°C.

[0172] For example, a pharmaceutical formulation may also be considered stable if, after 28 days of storage at 40°C, approximately 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, or 98% of native antibodies can be detected by SE-HPLC. A pharmaceutical formulation may also be considered stable if, after 3 months of storage at -20°C, approximately 96%, 97%, or 98% of native antibodies can be detected by SE-HPLC. A pharmaceutical formulation may also be considered stable if, after 3 months of storage at -30°C, approximately 96%, 97%, or 98% of native antibodies can be detected by SE-HPLC. A pharmaceutical formulation may also be considered stable if, after 3 months of storage at -80°C, approximately 96%, 97%, or 98% of native antibodies can be detected by SE-HPLC.

[0173] Stability can be measured, in particular, by determining the percentage of antibodies that form aggregates in the formulation after storage at a specified temperature for a specified time, and stability is inversely proportional to the percentage of aggregates formed. The percentage of aggregated antibodies can be determined, in particular, by size exclusion chromatography (e.g., size exclusion high-performance liquid chromatography [SE-HPLC]). Where the term is used herein, “acceptable stability” means that, after storage at a given temperature for a specified time, up to 6% of the antibodies are in a detectable aggregated form in the formulation. In certain embodiments, acceptable stability means that, after storage at a given temperature for a specified time, up to approximately 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the antibodies can be detected in aggregates in the formulation. The specified time period until stability is measured may be at least two weeks, at least 28 days, at least one month, at least two months, at least three months, at least four months, at least five months, at least six months, at least seven months, at least eight months, at least nine months, at least ten months, at least eleven months, at least twelve months, at least eighteen months, at least twenty-four months, or longer. The temperature at which the pharmaceutical formulation may be stored when assessing stability may be any temperature between approximately -80°C and approximately 40°C, for example, approximately -80°C, approximately -65°C, approximately -30°C, approximately -20°C, approximately 0°C, approximately 4°C to 8°C, approximately 5°C, approximately 25°C, approximately 35°C, approximately 37°C, or approximately 40°C.

[0174] For example, a pharmaceutical formulation may be considered stable if, after 6 months of storage at 5°C, approximately 3%, 2%, 1%, 0.5%, or 0.1% of antibodies are detected in agglutinated form. A pharmaceutical formulation may also be considered stable if, after 6 months of storage at 25°C, approximately 4%, 3%, 2%, 1%, 0.5%, or 0.1% of proteins are detected in agglutinated form. A pharmaceutical formulation may also be considered stable if, after 28 days of storage at 40°C, approximately 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of antibodies are detected in agglutinated form. A pharmaceutical formulation may also be considered stable if, after 3 months of storage at -20°C, -30°C, approximately -65°C, or -80°C, approximately 3%, 2%, 1%, 0.5%, or 0.1% of antibodies are detected in agglutinated form.

[0175] Stability can also be measured, among other things, by determining the proportion of antibody that migrates in the more acidic fraction during ion exchange ("acidic form") rather than in the main fraction of antibody ("main charge form"), and stability is inversely proportional to the fraction of antibody in the acidic form. While we do not wish to be bound by theory, deamidation of an antibody can make the antibody more negatively charged and therefore more acidic than an undeamidated antibody (e.g., Robinson, N., Protein Deamidation, PNAS, April 16, 2002, 99(8):5283-5288). The proportion of "acidified" antibody can be determined, among other things, by ion exchange chromatography (e.g., cation exchange high-performance liquid chromatography [CEX-HPLC]) or imaging capillary isoelectric focusing (iCIEF). Where the term is used herein, "acceptable stability" means that, after storage at a defined temperature for a defined time, up to 49% of the antibody is in the more acidic form detected in the formulation. In certain embodiments, acceptable stability means that, after storage at a given temperature for a specified time, up to approximately 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the antibody can be detected in the formulation in an acidic form. The specified amount of time until stability is measured may be at least two weeks, at least 28 days, at least one month, at least two months, at least three months, at least four months, at least five months, at least six months, at least seven months, at least eight months, at least nine months, at least ten months, at least eleven months, at least twelve months, at least eighteen months, at least twenty-four months, or longer. When evaluating stability, pharmaceutical formulations can be stored at any temperature between approximately -80°C and approximately 40°C, for example, approximately -80°C, approximately -65°C, approximately -30°C, approximately -20°C, approximately 0°C, approximately 4°C to 8°C, approximately 5°C, approximately 25°C, or approximately 40°C.

[0176] For example, a pharmaceutical formulation may be considered stable after 3 months of storage at -80°C, -30°C, or -20°C if approximately 30%, 29%, 28%, 27%, 26%, 25%, 24%, 23%, 22%, 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the antibodies are in a more acidic form. Pharmaceutical formulations may also be considered stable after 6 months of storage at 5°C if approximately 32%, 31%, 30%, 29%, 28%, 27%, 26%, 25%, 24%, 23%, 22%, 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the antibodies are in a more acidic form. Pharmaceutical formulations may also be considered stable after 6 months of storage at 25°C if approximately 43%, 42%, 41%, 40%, 39%, 38%, 37%, 36%, 35%, 34%, 33%, 32%, 31%, 30%, 29%, 28%, 27%, 26%, 25%, 24%, 23%, 22%, 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the antibodies are in a more acidic form. Pharmaceutical formulations may also be considered stable after 28 days of storage at 40°C if approximately 49%, 48%, 47%, 46%, 45%, 44%, 43%, 42%, 41%, 40%, 39%, 38%, 37%, 36%, 35%, 34%, 33%, 32%, 31%, 30%, 29%, 28%, 27%, 26%, 25%, 24%, 23%, 22%, 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the antibodies are in a more acidic form.

[0177] Other methods, such as differential scanning calorimetry (DSC) to determine thermal stability, controlled stirring to determine mechanical stability, and absorbance at approximately 350 nm or approximately 405 nm to determine solution turbidity, may be used to evaluate the stability of the formulations of this disclosure. For example, the formulations of this disclosure have been stored at approximately 5°C to approximately 25°C for 6 months or more, and their OD (Oxidation Disorder) 405 The change in the formulation at point zero is the OD 405 A value less than approximately 0.05 (for example, 0.04, 0.03, 0.02, 0.01, or less) may be considered stable.

[0178] Measuring the physiological activity or binding affinity of an antibody to its target can also be used to assess stability. For example, a formulation of this disclosure may be considered stable if, after storage for a specified time (e.g., 1 to 12 months) at, for example, 5°C, 25°C, or 40°C, the anti-hOX40L antibody contained in the formulation binds to hOX40L with an affinity of at least 90%, 95%, or higher than the binding affinity of the antibody before storage. Binding affinity can be determined, for example, by ELISA or plasmon resonance. Biological activity can be determined, for example, by an hOX40L activity assay, such as contacting cells expressing hOX40L with a formulation containing the anti-hOX40L antibody. Binding of the antibody to such cells can be measured directly, for example, by FACS analysis.

[0179] Alternatively, the downstream activity system of hOX40L can be measured in the presence of an antibody and compared to the activity system of hOX40L in the absence of an antibody. In some embodiments, hOX40L may be endogenous to cells.

[0180] In one embodiment, at least 91% of the antibodies in the aqueous pharmaceutical formulation have a native conformation after 28 days at 40°C, as measured by size exclusion chromatography (SE-HPLC). In one embodiment, at least 94% of the antibodies in the aqueous pharmaceutical formulation have a native conformation after 8 weeks at 25°C, as measured by size exclusion chromatography. In one embodiment, at least 98% of the antibodies in the aqueous pharmaceutical formulation have a native conformation after 16 weeks at 5°C, as measured by size exclusion chromatography. In one embodiment, at least 98% of the antibodies in the aqueous pharmaceutical formulation have a native conformation after 16 weeks at -65°C, as measured by size exclusion chromatography. When measured by size exclusion chromatography, the aqueous pharmaceutical formulation contains a major peak species, a high molecular weight (HMW) species, and a low molecular weight (LMW) species. As used herein, “native conformation” refers to the major peak species as measured by SE-HPLC.

[0181] In one embodiment, at least 55% of the antibody in the formulation disclosed herein is the major charge variant after 28 days at 40°C, as measured by imaging capillary isoelectric focusing (iCIEF). In one embodiment, at least 65% of the antibody in the formulation disclosed herein is the major charge variant after 16 days at 25°C, as measured by imaging capillary isoelectric focusing (iCIEF). In one embodiment, at least 70% of the antibody in the formulation disclosed herein is the major charge variant after 16 days at 5°C, as measured by imaging capillary isoelectric focusing (iCIEF). In one embodiment, at least 70% of the antibody in the formulation disclosed herein is the major charge variant after 16 days at -65°C, as measured by imaging capillary isoelectric focusing (iCIEF).

[0182] In some embodiments, the aqueous pharmaceutical formulation is stable upon freezing and thawing. In some embodiments, the aqueous pharmaceutical formulation is stable when stored at -65°C, 5°C, or 25°C for at least 16 weeks. In some embodiments, the aqueous pharmaceutical formulation is stable when stored at 40°C for at least 8 weeks. In some embodiments, the aqueous pharmaceutical formulation is stable when stored at 5°C for at least 1 year. In some embodiments, the aqueous pharmaceutical formulation is contained in a suitable container to maintain the above-mentioned stability.

[0183] Pharmaceutical dosage forms A unit dosage form of a formulated antibody that specifically binds to OX40L is provided. In various embodiments, the unit dosage form comprises the aqueous pharmaceutical formulation described herein and is suitable for parenteral administration to mammalian subjects. In some embodiments, the aqueous pharmaceutical formulation is suitable for intravenous, subcutaneous, or intramuscular administration. In some embodiments, the unit dosage form may be contained in a suitable container having a specific volume of the aqueous pharmaceutical formulation disclosed. In some embodiments, the unit dosage form may be contained in a container (e.g., a vial) so that a predetermined dose of the antibody can be removed from the container for administration to a patient.

[0184] In some embodiments, the unit dosage form contained in the container may contain about 2.4 mL of the disclosed formulation. In some embodiments, the extractable volume of the unit dosage form is about 2 mL of the disclosed formulation. In some embodiments, the extractable volume of the unit dosage form is about 1 mL of the disclosed formulation. In some embodiments, the unit dosage form is contained in a suitable container, such as a glass vial with a stopper or cap. In one embodiment, the suitable container is a syringe, in which the unit dosage form of the aqueous pharmaceutical formulation disclosed herein is pre-filled in the syringe. For example, the syringe may be a pre-filled syringe, which may include a safety system, such as a needle guard, such as BD UltraSafe® or BD UltraSafe Plus® needle guard. In another example, the syringe may be a pre-filled pen or auto-injector device. In some embodiments, the unit dosage form contained in the container (such as a vial) may contain about 2.4 mL of the formulation disclosed herein, thereby the unit dosage form contains 180 ± 27 mg to 331 ± 49.7 mg of antibody. In some embodiments, the unit dosage form contained in the container contains about 300 ± 45 mg of the antibody disclosed herein. In some embodiments, the unit dosage form contained in the container (e.g., a pre-filled syringe, a pre-filled syringe with a safety system, a pre-filled pen, or an auto-injector device) may contain about 1 mL of the formulation disclosed herein, thereby the unit dosage form contains about 125 mg of antibody. In some embodiments, the unit dosage form contained in the container (e.g., a pre-filled syringe, a pre-filled syringe with a safety system, a pre-filled pen, or an auto-injector device) may contain about 2 mL of the formulation disclosed herein, thereby the unit dosage form contains about 250 mg of antibody.

[0185] In some embodiments, the unit dosage forms of the formulated antibody are not lyophilized.

[0186] In some embodiments, the antibody, pharmaceutical formulation, and / or formulated antibody are contained in a glass vial fitted with an elastomer closure.

[0187] Packaged medicines and kits Packaged products In one embodiment, a pharmaceutical product is provided comprising the pharmaceutical product of the present disclosure in a container suitable for the storage and / or administration of the pharmaceutical product described herein. For example, the pharmaceutical product may be contained in a sealed and sterile plastic or glass container having a predetermined volume, such as a vial, ampoule, syringe, cartridge, or bottle. Different types of vials, including clear and opaque (e.g., amber) glass or plastic vials, may be used to contain the pharmaceutical product of the present disclosure. Similarly, any type of syringe may be used to contain or administer the pharmaceutical product of the present disclosure.

[0188] Specifically, the pharmaceutical formulations of this disclosure may be contained in “normal tungsten” or “low tungsten” syringes. As will be understood by those skilled in the art, the process of making glass syringes generally involves the use of a hot tungsten rod that functions to puncture the glass, thereby creating a hole from which liquid can be drawn out and discharged from the syringe. This process results in the deposition of trace amounts of tungsten on the inner surface of the syringe. Subsequent washing and other processing steps can be used to reduce the amount of tungsten in the syringe. As used herein, the term “normal tungsten” means that the syringe contains 500 ppb (parts per billion) or more of tungsten. The term “low tungsten” means that the syringe contains less than 500 ppb of tungsten. For example, the low-tungsten syringes according to this disclosure may contain tungsten in amounts of approximately 490, 480, 470, 460, 450, 440, 430, 420, 410, 390, 350, 300, 250, 200, 150, 100, 90, 80, 70, 60, 50, 40, 30, 20, or less than 10 ppb.

[0189] Rubber plungers used in syringes and rubber stoppers used to close the openings of vials may be coated to prevent contamination of the pharmaceutical contents of the syringe or vial, or to maintain their stability. Thus, the pharmaceutical formulations of this disclosure may, in certain embodiments, be contained within a syringe containing a coated plunger or within a vial sealed with a coated rubber stopper. For example, the plunger or stopper may be coated with a fluorocarbon film. Examples of coated stoppers or plungers suitable for use with vials and syringes containing the pharmaceutical formulations of this disclosure are described, for example, in U.S. Patents 4,997,423, 5,908,686, 6,286,699, 6,645,635, and 7,226,554, the contents of which are incorporated herein by reference in their entirety. Certain exemplary coated rubber stoppers and plungers that may be used in the context of this disclosure are commercially available under the trade name "FLUROTEC®" from West Pharmaceutical Services, Inc. (Lionville, PA). FLUROTEC® is an example of a fluorocarbon coating used to minimize or prevent pharmaceuticals from adhering to rubber surfaces.

[0190] According to certain embodiments of the present disclosure, the pharmaceutical formulation may be contained within a low-tungsten syringe comprising a fluorocarbon-coated plunger. The pharmaceutical formulation may be administered to a patient by parenteral routes such as injection (e.g., subcutaneous, intravenous, intramuscular, intraperitoneal, etc.), or by transdermal, mucosal, nasal, pulmonary, or oral administration. The pharmaceutical formulations of the present disclosure can be delivered subcutaneously using a variety of reusable pen or automated injector delivery devices. Examples include AUTOPEN (trademark) (Owen Mumford, Inc., Woodstock, UK), DISETRONIC (trademark) pen (Disetronic Medical Systems, Bergdorf, Switzerland), HUMALOG MIX75 / 25 (trademark) pen, HUMALOG (trademark) pen, HUMALIN 70 / 30 (trademark) pen (Eli Lilly and Co., Indianapolis, IN), NOVOPEN (trademark) I, II, and III (Novo Nordisk, Copenhagen, Denmark), NOVOPEN JUNIOR (trademark) (Novo Nordisk, Copenhagen, Denmark), BD (trademark) pen (Becton Dickinson, Franklin Lakes, NJ), OPTIPEN (trademark), OPTIPEN PRO (trademark), OPTIPEN Examples include, but are not limited to, STARLET (trademark) and OPTICLIK (trademark) (Sanofi-Aventis, Frankfurt, Germany).Examples of disposable pen or auto-injector delivery devices applicable to the subcutaneous delivery of the pharmaceutical formulations of this disclosure include SOLOSTAR® pen (Sanofi-Aventis), FLEXPEN® (Novo Nordisk), and KWIKPEN® (Eli Lilly), SURECLICK® auto-injector (Amgen, Thousand Oaks, CA), PENLET® (Haselmeier, Stuttgart, Germany), EPIPEN® (Dey, LP), HUMIRA® pen (Abbott Labs, Abbott Park, IL), MOLLY® auto-injector (SHL Medical AG, Zug, Switzerland), YPSOMATE auto-injector (Ypsomed AG, Burgdorf, Switzerland), BD PHYSIOJECT® auto-injector or (Becton, Dickinson and Examples include, but are not limited to, the REFLEX auto-injector (Sanofi-Aventis), as well as the REFLEX auto-injector (Sanofi-Aventis).

[0191] The use of microinfusers for delivering the pharmaceutical formulations of this disclosure is also contemplated herein. As used herein, the term “microinfuser” means a subcutaneous delivery device designed to slowly administer a large volume (e.g., up to approximately 2.5 mL or more) of a therapeutic formulation over an extended period (e.g., approximately 10, 15, 20, 25, 30 minutes or more). See, for example, U.S. Patent No. 6,629,949, U.S. Patent No. 6,659,982, and Meehan et al., J. Controlled Release 46:107-1 16 (1996). Microinfusers are particularly useful for delivering large volumes of therapeutic proteins contained in high-concentration (e.g., approximately 100, 125, 150, 175, 200 mg / mL or more) solutions or viscous solutions.

[0192] More examples of suitable containers and packaged drugs, including aqueous formulations disclosed herein, are listed below. • Glass vials containing an aqueous formulation comprising an anti-OX40L antibody or its antigen-binding fragment. A drug delivery device containing an aqueous formulation comprising an anti-OX40L antibody or its antigen-binding fragment. • A pre-filled syringe containing an aqueous formulation comprising an anti-OX40L antibody or its antigen-binding fragment. The pre-filled syringe may have a safety system, such as a passive needle guard (e.g., BD UltraSafe® or BD UltraSafe Plus® needle guard). The pre-filled syringe may contain an aqueous formulation comprising an anti-OX40L antibody or its antigen-binding fragment in a volume of up to 1 mL or up to 2 mL. In one embodiment, the pre-filled syringe contains about 125 mg of amliterimab in a 1 mL solution (125 mg / mL ± 18.75 mg / mL). In one embodiment, the pre-filled syringe contains about 250 mg of amliterimab in a 2 mL solution (125 mg / mL ± 18.75 mg / mL). • Microinfuser containing an aqueous formulation containing an anti-OX40L antibody or its antigen-binding fragment. A pen delivery device containing an aqueous formulation comprising an anti-OX40L antibody or its antigen-binding fragment. The pen delivery device may be a reusable pen delivery device. The pen delivery device may be a disposable pen delivery device. The pen delivery device may contain an aqueous formulation comprising an anti-OX40L antibody or its antigen-binding fragment in a volume of up to 1 mL or up to 2 mL. In one embodiment, the pen delivery device contains 125 mg of amliterimab in 1 mL of solution (125 mg / mL ± 18.75 mg / mL). In one embodiment, the pen delivery device contains 250 mg of amliterimab in 2 mL of solution (125 mg / mL ± 18.75 mg / mL). • An automated syringe delivery device containing an aqueous formulation comprising an anti-OX40L antibody or its antigen-binding fragment. The automated syringe delivery device may contain an aqueous formulation comprising an anti-OX40L antibody or its antigen-binding fragment in a volume of up to 1 mL or up to 2 mL. In one embodiment, the automated syringe delivery device contains 125 mg of amliterimab in 1 mL of solution (125 mg / mL ± 18.75 mg / mL). In one embodiment, the automated syringe delivery device contains 250 mg of amliterimab in 2 mL of solution (125 mg / mL ± 18.75 mg / mL).

[0193] Exemplary drug delivery devices may include needle-based injection systems, such as those described in Table 1 of Section 5.2 of ISO 11608-1:2014(E). As described in ISO 11608-1:2014(E), needle-based injection systems can be broadly distinguished into multi-dose container systems and single-dose (with partial or complete emptiness) container systems. Containers may be replaceable or integrated, non-replaceable containers.

[0194] As further described in ISO 11608-1:2014(E), a multi-dose container system may include a needle-based injection device with replaceable containers. In such a system, each container holds a multi-dose, the size of which may be fixed or variable (pre-set by the user). Another multi-dose container system may include a needle-based injection device with an integrated, non-replaceable container. In such a system, each container holds a multi-dose, the size of which may be fixed or variable (pre-set by the user).

[0195] As further described in ISO 11608-1:2014(E), a single-dose container system may include a needle-based injection device with replaceable containers. In one example of such a system, each container holds a single dose, thereby discharging the entire deliverable volume (complete discharge). In a further example, each container holds a single dose, thereby discharging a portion of the deliverable volume (partial discharge). Also, as described in ISO 11608-1:2014(E), a single-dose container system may include a needle-based injection device with an integrated, non-replaceable container. In one example of such a system, each container holds a single dose, thereby discharging the entire deliverable volume (complete discharge). In a further example, each container holds a single dose, thereby discharging a portion of the deliverable volume (partial discharge).

[0196] An example of a sleeve-operated autoinjector with manual needle insertion is described in International Publication No. 2015 / 004052. An exemplary audible dose completion feedback mechanism is described in International Publication Nos. 2016 / 193346 and 2016 / 193348. An exemplary needle safety mechanism after use of an autoinjector is described in International Publication No. 2016 / 193352. An exemplary needle sheath removal mechanism for a syringe autoinjector is described in International Publication No. 2016 / 193353. An exemplary support mechanism for supporting the axial position of a syringe is described in International Publication No. 2016 / 193355.

[0197] In some embodiments, sealed containers containing aqueous pharmaceutical formulations disclosed herein are further provided. In some embodiments, the sealed container may be a vial, syringe, microinfuser, pen delivery device, or auto-injector containing the aqueous formulation disclosed herein in a volume of up to 1 mL, up to 2 mL, or up to 2.25 mL. In some embodiments, the glass injection vial is sealed with a rubber stopper or cap. In one embodiment, the container is a single or multi-chamber syringe.

[0198] In one embodiment, a pre-filled syringe is provided containing an aqueous pharmaceutical formulation at a pH of approximately 5.8 to approximately 6.2, comprising 125 mg / mL ± 18.75 mg / mL to 138 mg / mL ± 20.7 mg / mL of antibody or its antigen-binding fragment, 20 ± 3 mM L-histidine or histidine hydrochloride, 220 mM ± 33 mM sucrose, and 0.04% (w / v) ± 0.006% (w / v) polysorbate 80. The aqueous pharmaceutical formulation may have a volume of approximately 1 mL ± 0.15 mL to 2.4 mL ± 0.36 mL (e.g., 2 mL). In one embodiment, the syringe is a 1 mL long glass syringe filled with a 27 gauge thin-walled needle, a fluorocarbon-coated rubber plunger, and a rubber needle shield. In one embodiment, the syringe is an OMPI 1 mL glass syringe fitted with a 27 gauge needle, an FM27 rubber needle shield, and a 4023 / 50 rubber plunger coated with FLUROTEC®.

[0199] In one embodiment, a pre-filled syringe is provided containing an aqueous pharmaceutical formulation comprising approximately 62.5 mg / mL of an antibody or its antigen-binding fragment (e.g., amliterimab) at a pH of approximately 5.8 to approximately 6.2, preferably approximately 6, 10 mM ± 1.5 mM or approximately 10 mM L-histidine or histidine hydrochloride, 220 mM ± 33 mM or approximately 220 mM sucrose, approximately 10 μM EDTA, approximately 0.06% (w / v) polysorbate 80, and water. The pre-filled syringe may contain approximately 125 mg of the antibody or its antigen-binding fragment (e.g., amliterimab) in a 2 mL solution (62.5 mg / mL). Therefore, it is understood that the present invention provides a pre-filled syringe containing a 2 mL volume aqueous pharmaceutical formulation comprising about 62.5 mg / mL of an antibody or its antigen-binding fragment (e.g., amriterimab), about 10 mM L-histidine or histidine hydrochloride, about 220 mM sucrose, about 10 μM EDTA, about 0.06% (w / v) polysorbate 80, and water, at a pH of about 5.8 to about 6.2, preferably about 6.

[0200] In one embodiment, a pre-filled syringe is provided containing an aqueous pharmaceutical formulation comprising 125 mg / mL ± 18.75 mg / mL or about 125 mg / mL of antibody or its antigen-binding fragment (e.g., amliterimab), 10 mM ± 1.5 mM or about 10 mM of L-histidine or histidine hydrochloride, 220 mM ± 33 or about 220 mM of sucrose, about 10 μM of EDTA, about 0.06% (w / v) of polysorbate 80, and water, at a pH of about 5.8 to about 6.2, preferably about 6. The pre-filled syringe may contain about 250 mg of antibody or its antigen-binding fragment (e.g., amliterimab) in 2 mL of solution (125 mg / mL). Therefore, it is understood that the present invention provides a pre-filled syringe containing a 2 mL volume aqueous pharmaceutical formulation comprising about 125 mg / mL of an antibody or its antigen-binding fragment (e.g., amriterimab), about 10 mM L-histidine or histidine hydrochloride, about 220 mM sucrose, about 10 μM EDTA, about 0.06% (w / v) polysorbate 80, and water, at a pH of about 5.8 to about 6.2, preferably about 6.

[0201] In one embodiment, a pre-filled syringe is provided containing an aqueous pharmaceutical formulation comprising 125 mg / mL ± 18.75 mg / mL or about 125 mg / mL of antibody or its antigen-binding fragment (e.g., amriterimab), 10 mM ± 1.5 mM or about 10 mM of L-histidine or histidine hydrochloride, 220 mM ± 33 mM or about 220 mM of sucrose, about 0.06% (w / v) of polysorbate 80, about 10 μM of EDTA, and water, and including a safety system such as a passive needle guard (e.g., BD UltraSafe® or BD UltraSafe Plus® needle guard), wherein the pH of the aqueous pharmaceutical formulation is about 5.8 to about 6.2. The volume of the aqueous pharmaceutical formulation may be about 1 mL ± 0.15 mL to 2.4 mL ± 0.36 mL. In one embodiment, the syringe is a 1 mL glass syringe filled with a 27 gauge thin-walled needle, a fluorocarbon-coated rubber plunger, and a passive needle guard. In one embodiment, the syringe is an OMPI 1 mL glass syringe fitted with a 27 gauge needle, an FM27 rubber needle shield, and a FLUROTEC®-coated 4023 / 50 rubber plunger. In one embodiment, the pre-filled syringe contains 125 mg or approximately 125 mg of amliterimab in a 1 mL solution (125 mg / mL ± 18.75 mg / mL). Therefore, it is understood that the present invention provides a pre-filled syringe, including a safety system, containing a 1 mL volume aqueous pharmaceutical formulation containing about 125 mg / mL of an antibody or its antigen-binding fragment (e.g., amliterimab), about 10 mM L-histidine or histidine hydrochloride, about 220 mM sucrose, about 10 μM EDTA, about 0.06% (w / v) polysorbate 80, and water, at a pH of about 5.8 to about 6.2. In another embodiment, the pre-filled syringe contains 250 mg or about 250 mg of amliterimab in a 2 mL solution (125 mg / mL ± 18.75 mg / mL).Therefore, it is understood that the present invention provides a pre-filled syringe, including a safety system, containing a 2 mL volume aqueous pharmaceutical formulation containing about 125 mg / mL of an antibody or its antigen-binding fragment (e.g., amriterimab), about 10 mM L-histidine or histidine hydrochloride, about 220 mM sucrose, about 10 μM EDTA, about 0.06% (w / v) polysorbate 80, and water, at a pH of about 5.8 to about 6.2.

[0202] In one embodiment, a pre-filled pen or auto-injector device is provided containing an aqueous pharmaceutical formulation comprising 125 mg / mL ± 18.75 mg / mL or about 125 mg of an antibody or its antigen-binding fragment (e.g., amliterimab), 10 mM ± 1.5 mM or about 10 mM of L-histidine or histidine hydrochloride, 220 mM ± 33 mM or about 220 mM of sucrose, about 0.06% (w / v) of polysorbate 80, about 10 μM of EDTA, and water, wherein the pH of the aqueous pharmaceutical formulation is about 5.8 to about 6.2. The volume of the aqueous pharmaceutical formulation may be about 1 mL ± 0.15 ml to 2.4 ml ± 0.36 ml. In one embodiment, the pre-filled pen device contains 125 mg or about 125 mg of amliterimab in a 1 mL solution (125 mg / mL ± 18.75 mg / mL). Therefore, it is understood that the present invention provides a pre-filled pen or auto-injector device containing a 1 mL volume aqueous pharmaceutical formulation comprising, at a pH of about 5.8 to about 6.2, about 125 mg / mL of an antibody or its antigen-binding fragment (e.g., amliterimab), about 10 mM L-histidine or histidine hydrochloride, about 220 mM sucrose, about 10 μM EDTA, about 0.06% (w / v) polysorbate 80, and water. In another embodiment, the pre-filled pen device contains 250 mg or about 250 mg of amliterimab in a 2 mL solution (125 mg / mL ± 18.75 mg / mL). Therefore, it is understood that the present invention provides a pre-filled pen or auto-injector device containing a 2 mL volume aqueous pharmaceutical formulation comprising, at a pH of about 5.8 to about 6.2, about 125 mg / mL of an antibody or its antigen-binding fragment (e.g., amriterimab), about 10 mM L-histidine or histidine hydrochloride, about 220 mM sucrose, about 10 μM EDTA, about 0.06% (w / v) polysorbate 80, and water.

[0203] In some embodiments, pharmaceuticals comprising aqueous pharmaceutical formulations contained in a container, such as a pre-filled syringe or glass vial, have the stability attributes described above.

[0204] kit The present disclosure also provides a kit comprising the pharmaceutical formulations disclosed herein.

[0205] In some embodiments, the kit comprises a container such as the glass vial, drug delivery device, pre-filled syringe, microinfuser, pen delivery device, or autoinjector described above, containing the aqueous pharmaceutical formulation disclosed herein.

[0206] In some embodiments, the kit comprises a sealed container containing the aqueous pharmaceutical formulation described herein and at least one separate injection device for delivering the aqueous pharmaceutical formulation to a mammalian subject that needs it. In some embodiments, the injection device is a syringe, microinfuser, pen delivery device, or autoinjector. In some embodiments, the injection device is a single or multi-chamber syringe.

[0207] In certain exemplary embodiments, the kit of the present disclosure further comprises a control antibody that does not react with the hOX40L antigen. In certain embodiments, the kit of the present disclosure may contain substantially isolated hOX40L antigen as a control.

[0208] In some embodiments, the kit disclosed herein may further comprise a label or instructions containing a marketing approval number (e.g., FDA or EMA approval number).

[0209] In another specific embodiment, the kit of the present disclosure comprises means for detecting the binding of a modified antibody to hOX40L.

[0210] In one example, a kit is provided comprising an aqueous formulation, packaging, and instructions containing an anti-OX40L antibody, or an antigen-binding fragment thereof, for use in the treatment of atopic dermatitis.

[0211] In one embodiment, the kit contains the antibody of the Disclosure, for example, a purified antibody, in one or more containers and an antigen (for example, an antibody may be conjugated to a detectable substrate such as a fluorescent compound, an enzyme substrate, a radioactive compound, or a luminescent compound, or a second antibody that recognizes a first antibody may be conjugated to a detectable substrate). In a particular embodiment, the kit may contain a recombinantly produced or chemically synthesized hOX40L antigen. The hOX40L antigen provided in the kit may also be conjugated to a solid support. In a more specific embodiment, the detection means of the kit includes a solid support to which the hOX40L antigen is conjugated. Such a kit may also contain a non-adherent reporter-labeled anti-human antibody. In this embodiment, the binding of the antibody to the hOX40L antigen can be detected by the binding of the reporter-labeled antibody.

[0212] Preparation process Embodiments of this disclosure provide methods for preparing pharmaceutical formulations and unit dosage forms described herein.

[0213] In some embodiments, an antibody that specifically binds to OX40 ligand (OX40L) is expressed in a suitable cell culture. The antibody in the cell culture is typically subjected to at least one of a chromatography step and at least one ultrafiltration step to produce a purified antibody solution. The purified antibody solution may be adjusted to have a specific desired concentration of antibody, to add specific excipients, and to adjust the pH.

[0214] In one embodiment, a pharmaceutical formulation containing an anti-OX40L antibody formulated with the disclosed formulation may be prepared by 1) buffer exchange to achieve a target buffer concentration and pH, and subsequently 2) to achieve an upward concentration exceeding the target concentration.

[0215] In some embodiments, the antibody concentration is adjusted to a desired concentration in the range of about 28 mg / mL to about 138 mg / mL, and EDTA, sucrose, L-histidine or histidine hydrochloride, and polysorbate 80 are added, and the pH is adjusted to about 5.5 to about 6.5. In one embodiment, the antibody concentration is adjusted to about 28 mg / mL. In one embodiment, the antibody concentration is adjusted to about 31 mg / mL to about 125 mg / mL. In one embodiment, the antibody concentration is adjusted to about 31 mg / mL.

[0216] In some embodiments, the antibody concentration is adjusted to a range of approximately 75 mg / mL to approximately 138 mg / mL, sucrose, L-histidine or histidine hydrochloride, and polysorbate 80 are added, and the pH is adjusted to approximately 5.5 to approximately 6.5. In one embodiment, the antibody concentration is adjusted to 75 ± 11.25 mg / mL. In another embodiment, the antibody concentration is adjusted to 138 ± 20.7 mg / mL. In yet another embodiment, the antibody concentration is adjusted to 125 ± 18.75 mg / mL.

[0217] In some embodiments, the pH of the formulation is adjusted to about 5.5 to about 6.5, for example, 5.5 ± 0.2 to 6.5 ± 0.2. In some embodiments, the pH is adjusted to about 6, for example, 6.0 ± 0.2. In some embodiments, the pH is adjusted to 6.0. In some embodiments, the pH is adjusted to 6.2.

[0218] In some embodiments, the concentration of sucrose is about 220 mM, for example, 220 mM ± 33 mM; the concentration of EDTA is about 10 μM, for example, 10 μM ± 1.5 μM; the concentration of L-histidine or histidine hydrochloride is about 10 mM, for example, 10 mM ± 1.5 mM; and the concentration of polysorbate 80 is about 0.04% v / v, for example, 0.04% (w / v) ± 0.006% (w / v) or about 0.06% v / v, for example, 0.06% (w / v) ± 0.009% (w / v).

[0219] In some embodiments, the concentration of sucrose is about 220 mM, for example, 220 mM ± 33 mM; the concentration of L-histidine or histidine hydrochloride is about 20 mM, for example, 20 mM ± 3 mM; and the concentration of polysorbate 80 is about 0.04% v / v, for example, 0.04% (w / v) ± 0.006% (w / v).

[0220] The pharmaceutical formulations disclosed herein may be prepared from a bulk purified amliterimab active pharmaceutical ingredient (BPDS) in a solution containing a low concentration of amliterimab, such as about 10.1 g / L, in a buffer such as a buffer containing 25 mM L-histidine / L-histidine hydrochloride at pH 6.2. The bulk purified amliterimab in solution can be stored at 65°C or below from the date of manufacture until the materials are divided, compounded, and transferred to other suitable storage conditions.

[0221] In some embodiments, the formulated antibody is not lyophilized, and the pharmaceutical formulation is not in a lyophilized form.

[0222] In some embodiments, the formulated antibody is lyophilized, and the pharmaceutical formulation is in lyophilized form.

[0223] Protein concentration, pH value, and density are determined during BPDS processing and can be used for necessary calculations of batch formulations by using standard dilution procedures. The formulations can be compounded by spiking them into the target formulation of the bulk drug along with the excipient stock solution. The protein concentration, osmotic pressure, and pH of the final compounded solution can then be determined and verified.

[0224] In some embodiments, the prepared formulation solution may be filtered using a 0.22 μm polyvinylidene fluoride (PVDF) membrane filter.

[0225] In some embodiments, a formulation solution containing an antibody prepared according to the process described herein can be used as a primary packaging material and filled into a suitable container to produce a packaged pharmaceutical. In certain embodiments, the packaging material may be filled into a vial, such as a glass vial, observing aseptic techniques. In some embodiments, the container is a 6R / 20 mm Type I glass vial, the formulated solution is filled with a target fill volume of 2.4 mL, and then the vial is stoppered with a 20 mm bromobutyl rubber stopper (injection and lyophilization stopper) and sealed with a 20 mm aluminum flip-off seal.

[0226] In some other embodiments, the container is a syringe. In some embodiments, the syringe is a low tungsten glass syringe. In one embodiment, the syringe is a 1 mL long glass syringe from NUOVA OMPI with a 27G thin wall needle, a 4023 / 50 rubber stopper coated with FLUROTEC, and a tip cap made of FM27 rubber.

[0227] Use of Pharmaceutical Formulations The pharmaceutical formulations and packaged products of the present disclosure are suitable for administration to mammalian (human or non-human) subjects by parenteral routes such as subcutaneous, intravenous, intramuscular, intraperitoneal, or intradermal injection for treating, preventing, or ameliorating any disease or disorder related to OX40L activity, including diseases or disorders mediated by OX40L. Exemplary non-limiting diseases and disorders that can be treated or prevented by administration of the pharmaceutical formulations of the present disclosure include autoimmune disorders, inflammatory diseases or conditions, transplant rejection, and the like. In some embodiments, the pharmaceutical formulations and packaged products of the present disclosure are suitable for treating diseases such as asthma or atopic dermatitis.

[0228] Exemplary Embodiments [Table 1] [[ID=2I]]

[0229] In addition to their favorable characteristics in terms of stability and quality, the formulations FDS1.1 and FDS1.2 described above are safe and well-tolerated in humans. [Table 2]

[0230] The description of embodiments of this disclosure is not intended to be exhaustive or to limit the disclosure to the exact form disclosed. Specific embodiments and examples of the disclosure are described herein for illustrative purposes, but various equivalent modifications are possible within the scope of the disclosure, as will be recognized by those skilled in the art. For example, while method steps or functions are presented in a given order, alternative embodiments may perform the functions in a different order, or the functions may be performed substantially simultaneously. The teachings of the disclosure provided herein may be applied to other procedures or methods where appropriate. Further embodiments may be provided by combining the various embodiments described herein. Aspects of the disclosure may be modified, as needed, to adopt formulations, functions, and concepts of the above-mentioned references and uses in order to provide further embodiments of the disclosure. Furthermore, several modifications may be made in the protein structure without affecting the type or amount of biological or chemical action, taking into consideration biological functional equivalence. These and other modifications may be made to the detailed description disclosure. All such modifications are intended to be included within the scope of the appended claims.

[0231] Any particular element of any of the embodiments described above may be combined with or substituted for an element of another embodiment. Furthermore, while advantages associated with certain embodiments of this disclosure are described in the context of those embodiments, other embodiments may also demonstrate such advantages, and not all embodiments are required to demonstrate advantages that fall within the scope of this technology. Any part of this disclosure may be read in combination with any other part of this disclosure unless otherwise evident from the context. [Examples]

[0232] The following embodiments further illustrate aspects of the present disclosure. However, they do not in any way limit the teachings of the present disclosure described herein. It should be understood that these embodiments are provided for illustrative purposes only. From the above description and these embodiments, those skilled in the art can identify the essential features of embodiments of the present disclosure. Without departing from the spirit and scope thereof, those skilled in the art can make various changes and modifications to the present disclosure to suit various uses and conditions. All publications, including patents and non-patent literature, referenced herein are expressly incorporated herein by reference.

[0233] Abbreviation: ArgHCl: Arginine hydrochloride BDP: Bulk Pharmaceuticals BPDS: Bulk purified active pharmaceutical ingredient CE-SDS: Capillary electrophoresis of sodium dodecyl sulfate CAD: Charged Particle Detector cIEF: Capillary isoelectric focusing electrophoresis CR: Comparable to the standard DLS: Dynamic Light Scattering DP: Drug product DS: API DSC: Differential Scanning Calorimetry DTPA: Diethylenetriaminepentaacetic acid EDTA: Ethylenediaminetetraacetic acid ELISA: Enzyme-linked immunosorbent assay Eur.Ph.: European Pharmacopoeia FCC: Codex Alimentarius Commission on Food Chemistry FDS: Active pharmaceutical ingredient for formulation FIH: First in Human F / T: Freeze / Thaw GMP: Good Manufacturing Practice HDPE: High-density polyethylene HisHCl: Histidine hydrochloride iCIEF Imaging Capillary Isoelectric Focusing Electrophoresis iLAB LES Lab Execution System HIAC: High-precision Liquid Particle Counter HMWS: High Molecular Weight Species LC-MS: Liquid Chromatography-Mass Spectrometry LIMS Laboratory Information Management System LMWS: Low Molecular Weight Species <LoQ: Below the Quantification Limit of the Method NaCl: Sodium Chloride N / A: Not Applicable NBE: No Blowback NF: National Formulary NT = Not Tested NTU: Nephelometric Turbidity Unit OD: Optical Density PFVP: Practically Free of Visible Particles pI: Isoelectric Point PS80: Polysorbate 80 PVDF: Polyvinylidene Fluoride RP-HPLC: Reverse Phase High Performance Liquid Chromatography SC: Subcutaneous SEC: Size Exclusion Chromatography SE-HPLC: Size Exclusion High Performance Liquid Chromatography SL: Standard Line SOP: Standard Operating Procedure Tbd: Not Yet Available TM: Test Method TPP: Technical Pilot Plant UF / DF: Ultrafiltration / Diafiltration USP: United States Pharmacopeia PFS: Prefilled Syringe

[0234] In the following examples, amritelimab (also known as SAR445229 and KY1005) was used exemplarily as an antibody that specifically binds to hOX40L. Amritelimab is a fully human IgG4 kappa monoclonal antibody having an "IgG4-PE" constant region with Leu235Glu and Ser228Pro Fc mutations. Amritelimab comprises a VH domain containing the amino acid sequence described in SEQ ID NO: 34 and a VL domain containing the amino acid sequence described in SEQ ID NO: 48.

[0235] Example 1 - Development of an optimized and stable anti-hOX40L antibody formulation the purpose Amritelimab (also known as SAR445229 or KY1005) is a human anti-OX40L subclass G4PE kappa mAb that binds to OX40L to block its interaction with OX40, a major regulator of the immune system. Amritelimab is designed to rebalance the immune system by blocking the irregular activation and proliferation of pro-inflammatory effector T cells and promoting the expansion of anti-inflammatory regulatory T cells, without broad suppression of the immune system. Amritelimab is currently in clinical trials as a treatment for atopic dermatitis.

[0236] It is desirable that the new, stable formulation be appropriately usable at any dose, for example, approximately 62.5 mg to 550 mg, such as a 62.5 mg dose, or a 125 mg dose, a 250 mg dose, or a 500 mg dose. It is also desirable that it be presented as a drug with a shelf life of 24 months.

[0237] In this example, a series of comprehensive studies were conducted to develop an optimized anti-hOX40L formulation. These studies evaluated the stability, viability, and trends of the antibody formulation. The effects of buffer concentrations and pH, excipient types, polysorbate 80 (PS80) concentration, and metal chelating agents were all critically evaluated for the optimized development of aqueous amriterimab pharmaceuticals (DP) and bulk formulations (FDS).

[0238] Table 1-1 shows the preliminary quality target product profiles for the formulations and products to be developed. [Table 3]

[0239] material and method drug substance Amriterimab formulation drug substance (FDS) was prepared. The FDS concentration was generally greater than 138 mg / mL. For these studies in this example, the FDS was formulated to the target protein and excipient concentrations and filtered at 0.22 μm under laminar flow before sterile packing.

[0240] Excipients All excipients used during the formulation development research were United States Pharmacopeia (USP) and European Pharmacopoeia (Eur.Ph.) grade raw materials.

[0241] Inspection of particles visible to the naked eye Particles visible to the naked eye were analyzed by one or two analysts under the supervision of the visual inspection unit (Seidenader Maschinenbau GmbH). The DP containers were cleaned with lens paper to remove dust and fingerprints from the exterior before inspection.

[0242] PH measurement The pH of buffer and formulated mAb solutions was measured using Thermo-Scientific pH probes and meters. Results were considered comparable if the difference between repeated measurements was within 0.1 pH units.

[0243] Osmotic pressure measurement Osmotic pressure measurements were performed on 20 μL samples (n=2-3) using an osmotic pressure analyzer (Advanced Instruments, OsmoPRO). To ensure measurement accuracy, osmotic pressure standardization was performed before and after sample analysis.

[0244] Protein concentration measurement Total protein concentration was determined by measuring UV absorbance at 280 nm on a microfluidic chip of the Unchained Labs Big Lunatic system. Measurements were performed in duplicate on 2–5 μL samples.

[0245] Turbidity analysis Sample turbidity was quantified by measuring the optical density (OD) from 340 nm to 360 nm using a BioTek Synergy neo2 multimode reader. 200 μL of each sample was packed into a UV-Vis transparent 96-well plate. OD was determined as the average of the absorbance values ​​at 340 nm, 345 nm, 350 nm, 355 nm, and 360 nm.

[0246] SEC-HPLC for HMWS Agglutination analysis was performed by size exclusion chromatography (SEC). Samples were run on a 1260 series HPLC (Agilent, Santa Clara, CA) equipped with a TSK-GEL G3000SWXL (Tosoh Bioscience, Tokyo, Japan) analytical column and a matching guard column. The mobile phase used was 40 mM phosphate and 150 mM sodium chloride (pH 7.2) at a flow rate of 0.75 mL / min for 25 minutes. The protein concentration at injection was 5 mg / mL. Two injections were performed for each sample. Detection was performed by UV absorbance at 280 nm, and the relative percentage of each eluted species was determined by integrating the chromatographic peaks.

[0247] HIAC for particles invisible to the naked eye Particles invisible to the naked eye were measured by light shielding on a Beckman-Coulter High Precision Particle Counter (HIAC) Model 9703+. The system was flushed with MilliQ water until the particle count at 10 μm was less than 1 particle / mL. To ensure accurate particle concentration, standards at 2 μm, 10 μm, and 25 μm were measured, followed by a large wash to remove any background. Samples were measured in four separate 0.2 mL injections using the 0.8 mL method. The first sample measurement was ignored, and the next three were averaged.

[0248] Quantification of PS80 using HPLC / CAD PS80 was analyzed by HPLC-CAD. Samples were passed through a 1260 series HPLC (Agilent, Santa Clara, CA) equipped with a Water Oasis MAX column and a charged aerosol detector. The mobile phases used were 2% formic acid in water and 2% formic acid in isopropyl alcohol, at a flow rate of 1 mL / min for 8 minutes. Three injections were performed for each sample. Detection was performed by CAD, and the chromatographic peaks were integrated and the area under the curve compared to the calibration curve was determined to obtain the PS80 concentration.

[0249] Capillary gel electrophoresis Fragmentation analysis was performed by non-reducing capillary gel electrophoresis (NR-CE). Samples were run on a LabChip GXII instrument (PerkinElmer, Waltham, MA) using the supplier's Protein Clear HR chip, a pre-developed method, and a reagent kit. The electrophoretic peaks were aggregated to determine the percentages of monomers and low molecular weight species (LMWS). The monomer percentage was reported as purity percentage, and the LMWS percentage was reported as fragmentation percentage.

[0250] PTM quantification by LC-MS Protein sample solutions and reference standards were diluted to a concentration of 0.46 mg / mL using 6 M guanidine HCl and 250 mM Tris pH 8.5 (reducing / alkylation buffer; RAB). The initial concentrations were entered, and the dilution was calculated using Hamilton software with the following formula: Sample volume (μL) = 60 μL / Sample concentration (mg / mL) RAB volume (μL) = 130 μL - sample volume (μL)

[0251] Trypsin / Lys-C digestion of the samples was performed using a digestion buffer solution of 25 mM Tris, 20 mM methionine, and pH 7.1. For each sample, 25 μL of the digest was injected onto a C18 column for LC-MS analysis using a trifluoroacetate buffer system. The samples were analyzed using Q Exactive Full MS.

[0252] The generated LC-MS data was processed using Chromeleon 7.2.10 software for relative quantification of modifications, including Met / Trp oxidation, deamidation, Asp isomerization, and HC C-terminal modifications, as well as for glycan and charge analysis.

[0253] cIEF of charge variants Protein charge heterogeneity was measured by capillary isoelectric focusing (cIEF) using a Protein Simple Maurice instrument in fluorescence detection mode with a 10-second exposure. Samples and standard solutions were first diluted to 5 mg / mL in water, and then to 0.5 mg / mL in water. Samples were mixed with MasterMix using the onboard mixer before analysis. Results were considered comparable if the difference was ≤10%.

[0254] Buffer / Backend Screening This study was conducted to evaluate the effects of buffer identity and pH on the physical and chemical stability of amriterimab aqueous formulations at low and high concentrations during PFS under refrigerated, accelerated, and stress storage conditions. The study also monitored the stability of high-concentration aqueous formulations in vials stored at cryogenic storage temperatures. A histidine buffer system with a pH of 5.5–6.3 was selected for this study.

[0255] Sample conditions To encompass amliterimab DP concentrations of 31–125 mg / mL, amliterimab concentrations of 28 mg / mL and 138 mg / mL were selected, including a 10% variability in concentration. Preparations containing 10 mM or 20 mM histidine buffer at a pH in the range of 5.5–6.3, along with 220 mM sucrose, 0.04% PS80, and optionally 10 μM EDTA, were prepared and stored in OMPI syringes and analyzed as detailed below. [Table 4]

[0256] Visual examination All formulations containing 28 mg / mL in syringe reference numbers 1_s to 6_s, and all formulations containing 138 mg / mL of amliterimab in syringe reference numbers 11_s to 16_s, were observed to remain visually clear and colorless after 12 weeks of storage in OMPI syringes at 5°C, 25°C, and 40°C.

[0257] Aggregation by SEC-HPLC Aggregation of formulations contained in syringes reference numbers 1_s~6_s and 11_s~16_s was evaluated by SEC-HPLC. Figures 1A, 1B, and 1C graphically show the change in HMWS of formulations contained in OMPI syringes after storage at 5°C, 25°C, and 40°C for 12 weeks, respectively. As shown in Figures 1A, 1B, and 1C, all of the exemplary formulations studied showed an increase in HMWS% in all formulations stored at 40°C, but did not demonstrate a substantial increase in HMWS% after 12 weeks of storage at 5°C and 25°C. In general, samples with higher protein concentrations tended to aggregate more than samples with lower protein concentrations. While there are some slight differences in the tendency for HMWS formation between different formulations, overall, samples from all formulations exhibit less than 1% HMWS after 12 weeks at 5°C, less than 2% HMWS after 12 weeks at 25°C, and less than 5% HMWS after 12 weeks at 40°C.

[0258] HIAC: Particles invisible to the naked eye Invisible particles in the exemplary formulations contained in syringe reference numbers 1_s~6_s and 11_s~16_s were evaluated by HIAC. Figures 2A, 2B, and 2C graphically show the changes in invisible particles larger than 2 μm in the exemplary formulations contained in OMPI syringes when stored for 12 weeks at 5°C (Figure 3A), 25°C (Figure 2B), and 40°C (Figure 2C). Figures 3A, 3B, and 3C graphically show the changes in invisible particles larger than 10 μm in the exemplary formulations when stored for 12 weeks at 5°C (Figure 3A), 25°C (Figure 3B), and 40°C (Figure 3C). Figures 4A, 4B, and 4C graphically show the changes in invisible particles larger than 25 μm in an example formulation contained in an OMPI syringe when stored for 12 weeks at 5°C (Figure 4A), 25°C (Figure 4B), and 40°C (Figure 4C).

[0259] As shown in Figures 2-4, at all particle sizes, the formation of particles invisible to the naked eye was relatively more pronounced at a stress temperature of 40°C compared to 5°C and 25°C. At 40°C, all formulations showed an overall increase in particles over 12 weeks, and samples with low protein concentrations showed the generation of relatively larger particles than samples with high protein concentrations.

[0260] Turbidity analysis Plate-based UV measurements were performed to track the turbidity and opacity of all formulation samples in syringe reference numbers 1_s~6_s and 11_s~16_s during storage. The turbidity results measured for all formulation samples after 12 weeks of storage at 5°C, 25°C, and 40°C are shown graphically in Figures 5A, 5B, and 5C. Figure 5A shows the turbidity results for formulation samples stored at 5°C for 12 weeks, Figure 5B shows the turbidity results for formulation samples stored at 25°C for 12 weeks, and Figure 5C shows the turbidity results for formulation samples stored at 40°C for 12 weeks.

[0261] It was found that protein concentration and storage temperature affect turbidity. Similar trends were observed for both low-protein and high-protein samples in the formulation.

[0262] Surprisingly, formulations containing 20 mM histidine tended to have higher turbidity at the start, and this tendency was particularly pronounced under stress conditions at 40°C compared to formulations containing 10 mM histidine.

[0263] Purification and fragmentation by capillary electrophoresis Purity was measured by capillary electrophoresis, and for most samples, the monomer content in the formulation samples was monitored over 12 weeks of storage at 5°C, 25°C, and 40°C. The results are shown in Figures 6A, 6B, and 6C, with Figure 6A showing the results at 5°C, Figure 6B showing the results at 25°C, and Figure 6C showing the results at 40°C. As can be seen from Figures 6A, 6B, and 6C, the majority of the samples maintained a purity of over 90% over 12 weeks at both 5°C and 25°C. At 40°C, fragmentation was common, with a slightly larger increase in lower-concentration samples. The increase in LMWS% was found to be comparable between lower-concentration and higher-concentration samples.

[0264] Quantification of PS80 The quantification of PS80 in exemplary formulations stored in syringes with reference numbers 1-6 and 11-16 over 12 weeks at 5°C, 25°C, and 40°C was analyzed. The results are shown graphically in Figures 7A, 7B, and 7C. Figure 7A shows the quantification of PS80 in exemplary formulations stored in syringes over 12 weeks at 5°C, Figure 7B shows the quantification of PS80 in formulations over 12 weeks at 25°C, and Figure 7C shows the quantification of PS80 in formulations over 12 weeks at 40°C. In this study, the methodological variability of PS80 analysis by CAD was approximately 25% (±100 ppm PS80). No trend in the change of PS80 over time was observed at 5°C, 25°C, and 40°C, and the percentage change was within the range of analytical variability. At both low and high protein concentrations, 10 mM histidine pH 6.3 (syringe reference numbers 4_s and 14_s, respectively) showed an increasing percentage change at 4 and 8 weeks, while PS80 concentrations remained within the range of variability at 12 weeks. At all temperatures, there was no significant decrease in PS80 after 12 weeks in these formulations.

[0265] Charge variant Charge variants were analyzed by capillary isoelectric focusing (cIEF) for exemplary formulations stored at syringe reference numbers 1-6 and 11-16 over 8 weeks at 5°C, 25°C, and 40°C. The results are shown graphically in Figures 8A, 8B, and 8C, with Figure 8A showing the charge variants of formulations over 8 weeks of storage at 5°C, Figure 8B showing the charge variants of formulations over 8 weeks of storage at 25°C, and Figure 8C showing the charge variants of formulations over 8 weeks of storage at 40°C. The results indicate that all formulations had comparable variants at each time point. At 40°C, most formulations tested showed an increase in acidic species and a decrease in the major peak over 8 weeks, while basic species remained constant.

[0266] Peptide mapping The modification of antibodies in the formulations was measured by peptide mapping after storage at 40°C for 3 months. The weekly percentage change in deamidation at HC N332 and oxidation at HC M103 of the antibodies was calculated. The results are shown graphically in Figures 9 and 10, with Figure 9 showing deamidation at HC N332 and Figure 10 showing oxidation at HC M103. No significant differences in the weekly change in deamidation were observed across formulations or protein concentrations. In formulations with 25 mg / mL of protein, oxidation at HC M103 in the antibody showed a slightly higher weekly oxidation percentage compared to formulations with a higher protein concentration of 138 mg / mL.

[0267] pH The pH of the formulations was measured at t0 and after 12 weeks of storage at 25°C and 40°C. As shown in Figure 11, no significant changes in pH were observed for any formulation at any given time point, even under accelerated or stress storage conditions (25°C and 40°C) at 28 mg / mL or 138 mg / mL.

[0268] Osmotic pressure The osmotic concentration of the formulations was measured at t0 and over 12 weeks of storage at 40°C. As shown in Figure 12, no significant changes in osmotic concentration were observed for any formulation up to 12 weeks under stress conditions at 40°C, 28 mg / mL, or 138 mg / mL.

[0269] Effect of buffer on freeze stability The effect of buffer / pH on the freeze stability of mAbs after storage was evaluated for solutions in 6R Schott vials, reference numbers 11_v to 16_v, which contained 138 mg / mL of mAbs in buffer with 10 mM or 20 mM histidine, with pH varying from 5.5 to 6.3. Freeze stability from t0 after storage at -30°C and -80°C for up to 12 weeks was analyzed for HMWS%, turbidity, pH, concentration, and microscopic particles (≥2 μm, ≥10 μm, ≥25 μm).

[0270] The results indicate that freeze stability data was comparable for samples between different formulations stored at -30°C and -80°C. No significant changes in aggregation, turbidity, pH, concentration, or particle formation were observed for up to 12 weeks.

[0271] Stabilizer research Stabilizer studies were conducted to evaluate the effects of different stabilizers or extenders on the physical and chemical stability of amliterimab aqueous formulations at low and high concentrations during PFS under refrigerated, accelerated, and stress storage conditions. This study also monitored the stability of high-concentration aqueous formulations in vials stored at freezing temperatures.

[0272] Sample conditions Aqueous formulations containing 28 mg / mL or 138 mg / mL of amliterimab, with a 10% variation in concentration, were prepared according to the components and concentrations listed in Tables 1-3. Such prepared aqueous formulations were filled into OMPI syringes. [Table 5]

[0273] Visual examination All formulations containing 28 mg / mL in syringe reference numbers 1_s, 7_s, and 8_s, as well as all formulations containing 138 mg / mL of amriterimab in syringe reference numbers 11_s, 17_x, and 18_s, were observed to remain visually clear and colorless after 12 weeks at 5°C, 25°C, and 40°C.

[0274] Aggregation by SEC-HPLC Aggregation of the formulations in syringe reference numbers 1_s, 7_s, 8_s, 11_s, 17_x, and 18_s was evaluated by SEC-HPLC. Figures 13A, 13B, and 13C graphically show the changes in HMWS of the formulations contained in these syringes over 12 weeks of storage at 5°C (Figure 13A), 25°C (Figure 13B), and 40°C (Figure 13C). The results show a general increase in HMWS% over 12 weeks of storage at 40°C, particularly for high-concentration samples, but there were no significant differences between formulations.

[0275] HIAC: Particles invisible to the naked eye Invisible particles in these exemplary formulations with syringe reference numbers s, 7_s, 8_s, 11_s, 17_x, and 18_s were evaluated by HIAC. Figures 14A, 14B, and 14C graphically show the changes in invisible particles larger than 2 μm in the exemplary formulations over 12 weeks of storage at 5°C (Figure 14A), 25°C (Figure 14B), and 40°C (Figure 14C). Figures 15A, 15B, and 15C graphically show the changes in invisible particles larger than 10 μm in the exemplary formulations over 12 weeks of storage at 5°C, 25°C, and 40°C, respectively. Figures 16A, 16B, and 16C graphically show the changes in invisible particles larger than 25 μm in the exemplary formulations over 12 weeks of storage at 5°C, 25°C, and 40°C, respectively.

[0276] At all particle sizes, a relatively significant increase in the formation of particles invisible to the naked eye was observed at a stress temperature of 40°C compared to 5°C and 25°C. At 40°C, all formulations showed an overall increase in particles over 12 weeks, and samples with lower protein concentrations showed the generation of relatively larger particles than samples with higher protein concentrations.

[0277] Turbidity analysis Turbidity analysis was performed on sample formulations stored in syringe reference numbers s, 7_s, 8_s, 11_s, 17_x, and 18_s at t0, and after storage for 2 weeks, 4 weeks, 8 weeks, and / or 12 weeks at 5°C, 25°C, and 40°C. The results are shown in Figures 17A, 17B, and 17C, which represent the results for storage at 5°C, 25°C, and 40°C, respectively.

[0278] The results indicate that turbidity values ​​for all stabilizer formulations were dependent on protein concentration and storage temperature. Regarding formulation trends, high-protein concentration samples containing 120 mM sucrose and 50 mM arginine hydrochloride exhibited the highest turbidity among the tested samples. All low-protein concentration formulations were comparable across all temperatures.

[0279] Quantification of PS80 The PS80 quantification of formulations stored in syringes at 5°C, 25°C, and 40°C was analyzed at T0 and 12 weeks later. In this study, the variability of the CAD-based PS80 analysis method was approximately 25% (±100 ppm PS80). The results are shown graphically in Figures 18A, 18B, and 18C. No trend in the change of PS80 over time was observed at 5°C, 25°C, and 40°C, and the percentage change was within the range of analytical variability. At all temperatures, there was no significant decrease in PS80 in these formulations after 12 weeks.

[0280] Peptide mapping Modifications were measured by peptide mapping after storage at 40°C for 3 months. Percentage changes per week were calculated for antibody deamidation at HC N332 and oxidation at HC M103. The results are shown graphically in Figures 19 and 20, where Figure 19 shows deamidation at HC N332 and Figure 20 shows antibody oxidation at HC M103. The results indicated no significant differences in the weekly change in deamidation across formulation or protein concentrations.

[0281] Effect of stabilizers on freeze stability The effect of stabilizers on freeze stability was evaluated for solutions in 6R Schott vials with vial reference numbers 11_v, 17_v, and 18_v, which contain 138 mg / mL of mAb. Freeze stability from t0 after storage at -30°C and -80°C for up to 12 weeks was analyzed for HMWS%, turbidity, pH, concentration, and microscopic particles (≥2 μm, ≥10 μm, ≥25 μm).

[0282] The results showed that the freeze stability data was comparable for stabilizer samples stored at -30°C and -80°C. No significant changes in aggregation, turbidity, pH, concentration, or particle formation were observed for up to 12 weeks.

[0283] Syringe comparability study Studies were conducted to evaluate the effects of OMPI EZ-filled syringes and BD Neopak syringes on the physical and chemical stability of amriterimab aqueous formulations at low and high concentrations under refrigerated, accelerated, and stress storage conditions.

[0284] Sample conditions Pre-filled OMPI or BD syringes containing aqueous formulations of amliterimab at concentrations of 28 mg / mL or 138 mg / mL with a 10% variability in concentration were prepared according to the components and component concentrations listed in Tables 1-4. [Table 6]

[0285] Visual examination In both OMPI and BD syringes, all formulations containing 28 mg / mL and all formulations containing 138 mg / mL of amriterimab remained visually clear and colorless after 12 weeks at 5°C, 25°C, and 40°C. There were no visual differences between the OMPI and BD syringes.

[0286] Aggregation by SEC-HPLC Aggregation of formulations in both OMPI and BD syringes was evaluated by SEC-HPLC. Figures 21A, 21B, and 21C graphically show the changes in HMWS of formulations in syringes 1_s_Omp, 9_s_BD, 10_s_BD, and 11_s_Omp over 12 weeks of storage at 5°C (Figure 21A), 25°C (Figure 21B), and 40°C (Figure 21C). The results show that the HMWS levels of each formulation in OMPI or BD syringe remained almost unchanged for 12 weeks of storage at 5°C and 25°C, and increased slightly over 12 weeks of storage at 40°C. There were no significant differences in HMWS between samples stored in OMPI and BD syringes.

[0287] HIAC: Particles invisible to the naked eye Invisible particles in these exemplary formulations in both OMPI and BD syringes were evaluated by HIAC. Figures 22A, 22B, and 22C graphically show the concentrations of invisible particles 2 μm or larger in OMPI and BD syringes over 12 weeks of storage at 5°C, 25°C, and 40°C, respectively. Figures 23A, 23B, and 23C graphically show the concentrations of invisible particles 10 μm or larger in exemplary formulations over 12 weeks of storage at 5°C, 25°C, and 40°C, respectively. Figures 24A, 24B, and 24C graphically show the concentrations of invisible particles 25 μm or larger in exemplary formulations over 12 weeks of storage at 5°C, 25°C, and 40°C, respectively.

[0288] The results show that, at 5°C and 25°C, the number of particles invisible to the naked eye increased in OMPI compared to BD syringes for both protein concentrations, across all particle sizes. Excess levels were observed only under stress conditions at 40°C in low-concentration samples in OMPI syringes.

[0289] Turbidity analysis Turbidity analysis was performed on the sample formulations at t0 and after 12 weeks of storage in syringes at 5°C, 25°C, and 40°C. The results are shown graphically in Figures 25A, 25B, and 25C, respectively. As shown in Figures 25A, 25B, and 25C, the high-protein concentration samples showed higher OD values ​​compared to the low-protein concentration samples, and the levels were comparable for formulations in both syringe types.

[0290] Quantification of PS80 The quantification of PS80 in formulations stored in syringes at 5°C, 25°C, and 40°C for 12 weeks was analyzed. In this study, the variability in PS80 analysis by CAD was approximately 25% (±100 ppm PS80). No trend in the change of PS80 over time was observed at 5°C, 25°C, and 40°C, and the percentage change was within the range of analytical variability. After 12 weeks at all temperatures, there was no significant decrease in PS80 in these formulations.

[0291] Charge variant Charge variants of the formulations were analyzed in both OMPI and BD syringes stored for 8 weeks at 5°C, 25°C, and 40°C. The results are shown graphically in Figures 26A, 26B, and 26C, respectively. The results show that at 5°C and 25°C (Figures 26A and 26B), the formulations in both syringes had comparable variants at each time point. At 40°C (Figure 26C), both OMPI and BD syringes at both protein concentrations showed an increase in acidic species and a decrease in the major peak over 8 weeks, while basic species remained relatively constant.

[0292] Peptide mapping Modifications were measured by peptide mapping after storage at 40°C for 3 months. Percent weekly change was calculated for antibody deamidation in exemplary formulations with HC N332 and antibody oxidation in exemplary formulations with HC M103. No significant differences in weekly change in deamidation were observed across formulations or protein concentrations (all were approximately 3% modification / week). No significant differences were observed across formulations with the same antibody concentration in OMPI or BD syringes; however, a higher percentage of oxidation at HC M103 was observed in formulations with lower antibody concentrations (i.e., 28 mg / mL) compared to formulations with 138 mg / mL of antibody. HC M103 oxidation % per week was approximately 0.35–0.43 for formulations with 28 mg / mL of antibody and approximately 0.23–0.26 for formulations with 138 mg / mL of antibody.

[0293] Chelating agent justification research This study was conducted to investigate the effects of transition metals, which may leach into DS during manufacturing, on the chemical and physical stability of the protein. In the current manufacturing process for amriterimab, the risk of transition metal contamination in DS and DP is low. However, if contamination is present, these transition metals can lead to chemical instability and aggregation in liquid solutions. Metal chelating agents can protect the chemical stability of proteins under metal exposure. EDTA and DTPA were evaluated for their ability to protect proteins during a series of worst-case experiments.

[0294] research design The study design is listed in Table 1-5. Aqueous formulations containing a base formulation with or without a chelating agent, comprising 125 mg / mL amriterimab, 10 mM histidine, 220 mM sucrose, and 0.04% PS80, were prepared according to the components and component concentrations listed in Table 1-5. Samples of the formulations were analyzed for visual appearance, aggregation, invisible particles, turbidity, PS80 quantification, peptide mapping, and the effect of stabilizers on freeze stability. 2R Schott vials were used in the study. [Table 7]

[0295] Visual examination No significant visual differences in turbidity or color were observed when comparing the formulations over time at all three temperatures.

[0296] Aggregation by SEC-HPLC Formulation aggregation was evaluated by analyzing the changes in sample formulations in syringes with reference numbers C-1 to C_6 ​​over 12 weeks of storage at 5°C, 25°C, and 40°C using SEC-HPLC. Figures 27A, 27B, and 27C graphically show the changes in HMWS% at 5°C (Figure 27A), 25°C (Figure 27B), and 40°C (Figure 27C).

[0297] At 5°C and 25°C (Figures 27A and 27B), no significant difference in HMWS% was found among formulations C-1 to C-6 for up to 12 weeks, although a slight increase in HMWS% over time was observed in all formulations at 25°C. As can be seen in Figure 27C, there was an overall increase in HMWS over 12 weeks of storage at 40°C, with the greatest increase in HMWS observed in formulations without chelating agents and containing ions at T12wk. The 10 μM DTPA formulation showed slightly lower HMWS at all time points compared to the 10 μM EDTA formulation. In the presence of chelating agents (EDTA or DTPA), ions did not significantly affect aggregation. Figure 27C further shows a comparison of the changes in HMWS in sample formulations over 2 months of storage at 40°C. As shown in Figure 27C, during storage at 40°C, a decrease of approximately 1–2% in HMWS% was observed when chelating agents were present in the formulations.

[0298] Quantification of PS80 The quantification of PS80 in formulations with reference numbers C-1 to C_6 ​​was analyzed. In this study, the variability of the CAD-based PS80 analysis method was approximately 25% (±100 ppm PS80). The results are shown in Figures 28A, 28B, 28C, and 28D. Figure 28A shows the PS80 (ppm) in the formulations over 2 months of storage at 5°C, 25°C, and 40°C, while Figures 28B, 28C, and 28D show the change in PS80 in formulations with reference numbers C_1 to C_6 ​​over 3 months of storage at 5°C (Figure 28B), 25°C (Figure 28C), and 40°C (Figure 28D). (Dotted lines indicate method variability).

[0299] As can be seen in Figure 28A, no significant decrease in PS80 was observed in these formulations after storage for 2 months at all temperatures. As can be seen in Figures 28B and 28D, no trend in the change of PS80 over time was observed at 5°C and 40°C, and the percentage change was within the range of analytical variability. At 25°C (Figure 28C), a moderate increasing trend over time was observed, but the change was at a level within the range of methodological variability. The highest PS80 loss was observed in formulations without a chelating agent, and the lowest decrease in PS80 was observed in formulations with 10 μM DTPA containing ions. After T12 wk at all temperatures, no significant decrease in PS80 was observed in these formulations.

[0300] Peptide mapping M103 oxidation and HC N332 deamidation were monitored by peptide mapping for all formulations stored in syringes at 40°C for up to 8 weeks. Percentage changes per week were calculated for deamidation at HC N332 (shown in Figure 29) and oxidation at HC M103 (shown in Figure 30). The results showed no significant differences across these formulations, regardless of the presence or absence of chelating agents. HC N332 deamidation measurements for antibodies in all exemplary formulations ranged from approximately 2.5 to 3.5, while HC M103 oxidation was approximately 0.25 to 0.5 in all exemplary formulations. The effect of chelating agents on deamination and oxidation was minimal.

[0301] Charge variant Charge variants were analyzed by CIEF at T0, T2wk (2 weeks), T4wk (4 weeks), and T8wk (8 weeks) for formulations with reference numbers C-1 to C_6, stored at 5°C, 25°C, and 40°C for 8 weeks. The results are shown in Figures 31A, 31B, and 31C, with acidic peaks shown in Figure 31A, basic peaks in Figure 31B, and the major peak in Figure 31C. The results indicate that all formulations had comparable variants at each time point tested. At 40°C, most formulations tested showed an increase in acidic species and a decrease in the major peak over 8 weeks, while basic species remained constant. The effect of chelating agents on charge variants was observed to be minimal.

[0302] Overall, based on this study, it is recommended to include 10 μM EDTA or DTPA in the formulation to provide mitigation against potential impurity leakage.

[0303] Justification of Polysorbate 80 This study was conducted to determine the optimal PS80 concentration required to stabilize the amriterimab aqueous formulation under agitation-induced stress.

[0304] As listed in Table 1-6, formulations F1-1 to F1-12 with different PS80 content were prepared. Various attributes of these formulations were analyzed at room temperature. This study consisted of three arms: List action: Samples were placed on a shaker for 1 hour, 3 hours, and 6 hours. Orbital shaking: Samples were placed on an orbital shaker at a speed of 300 rpm for 1 hour, 6 hours, and 3 days. List action + silicone oil: Samples with added silicone oil were placed on a list action shaker for 1 hour, 3 hours, and 6 hours. Schott 6R vials were used for this study. [Table 8]

[0305] List Action Research SEC-HPLC for HMWS All formulations subjected to the listing action were analyzed by SEC-HPLC for changes in HMWS at time points T0, T1hr, T3hr, and T6hr. Figures 32A, 32B, and 32C graphically show the measured changes in HMWS for formulations containing 31 mg / mL and 125 mg / mL of antibody over the listing action study period.

[0306] The results generally showed that HMW% increased with decreasing PS80% during the listing action. However, no substantial increase in HMWS during agitation was observed for any 31 mg / mL formulation with any PS80 concentration. HMWS levels were slightly higher for 31 mg / mL formulations with 0% and 0.02% PS80 compared to 31 mg / mL formulations with PS80 0.04%–0.14%.

[0307] A significant increase in HMWS was observed during agitation of a 125 mg / mL formulation with a 0.005% PS80 concentration, but this was not observed in the remaining 125 mg / mL formulations. Minimal change in HMW occurred when PS80 was present at a concentration of at least 0.02%. Therefore, a minimum PS80 concentration of 0.02% is considered necessary to protect against aggregation.

[0308] Turbidity analysis Turbidity analysis was performed on exemplary formulations containing 31 mg / mL and 125 mg / mL of antibodies at T0, T1hr, T3hr, and T6hr durations in the list action study. The results are shown in Figures 33A, 33B, and 33C.

[0309] As can be seen from Figures 33A, 33B, and 33C, in the absence of PS80, turbidity significantly increased for the 31 mg / mL formulation. No significant increase in turbidity was observed for the 31 mg / mL formulation with PS80 concentrations ranging from 0.02% to 0.14%. No significant increase in turbidity was observed for the 125 mg / mL formulation at any PS80 concentration. Based on turbidity data from the list-action study, a minimum PS80 concentration of 0.02% is required to minimize the increase in turbidity when exposed to agitation.

[0310] HIAC for particles invisible to the naked eye Invisible particles of at least 2 μm, at least 10 μm, or at least 25 μm in the exemplary formulations were measured by HIAC. The results are shown in Figures 34A to 34F, which graphically show the HIAC data for the 31 mg / mL and 125 mg / mL formulations during the List Action Study period.

[0311] As shown in Figures 34A to 34F, for the 31 mg / mL formulation, when the formulation was subjected to list-action shaking, there was a clear increase in particles larger than 2 μm over time in the absence of PS80. This was also observed in the 125 mg / mL formulation containing 0.005% PS80, even with particles larger than 2 μm.

[0312] Orbital shaking research SEC-HPLC for HMWS All formulations subjected to orbital shaking were analyzed by SEC-HPLC for changes in HMWS at T0, T1hr, T3hr, and T6hr. Figures 35A and 35B are graphs showing the results of HMWS changes for the 31 mg / mL and 125 mg / mL formulations over the orbital shaking study period.

[0313] As can be seen in Figures 35A and 35B, when subjected to orbital shaking, there was no significant increase in HMWS over time for any PS80 concentration, including the 31 mg / mL formulation. A significant increase in HMWS from T0 was observed for the 125 mg / mL formulation at a PS80 concentration of 0.005%, but the remaining 125 mg / mL formulations maintained a relatively constant HMWS% during shaking.

[0314] Turbidity analysis Turbidity analysis was performed on exemplary formulations subjected to orbital shaking at T0, T1hr, T3hr, and T6hr. The results show no significant increase in turbidity for the 31 mg / mL formulation with PS80 concentrations ranging from 0.04% to 0.14%. For the 125 mg / mL formulation with all PS80 concentrations, a significant increase in turbidity was observed at all three time points compared to T0, which may be due to instrumentation issues.

[0315] HIAC for particles invisible to the naked eye Invisible particles of at least 2 μm, at least 10 μm, or at least 25 μm in exemplary formulations subjected to orbital shaking were measured by HIAC. Figures 36A, 36B, 36C, 36D, 36E, and 36F are graphs showing HIAC data for 31 mg / mL and 125 mg / mL formulations during the orbital shaking period.

[0316] The results indicate that for the 125 mg / mL formulation with 0.005% PS80, relatively high particle counts were observed for particles larger than 2 μm, but not for particles of other sizes. The 31 mg / mL formulation with 0.04% PS80 showed slightly higher levels for particles larger than 2 μm and larger, and larger than 10 μm, compared to other formulations with the same protein concentration.

[0317] List Action + Silicone Oil Turbidity analysis was performed on exemplary formulations subjected to restorative stress + silicone oil at time points T0, T1hr, T3hr, and T6hr. The results are shown in Figures 37A and 37B, which are graphs showing turbidity data for 31 mg / mL and 125 mg / mL formulations over the study period.

[0318] During listing and silicone oil testing, no significant increase in turbidity was observed for 31 mg / mL formulations with various PS80 concentrations. A dramatic increase in turbidity was observed for a 125 mg / mL formulation with a PS80 concentration of 0.005%. No significant increase in turbidity was observed for 125 mg / mL formulations with PS80 concentrations ranging from 0.025% to 0.105%.

[0319] conclusion The objective of this formulation development activity was to identify the preferred concentration of PS80, as well as the type and concentration of chelating agent to be used. Additionally, pH / buffer screening was conducted to demonstrate DP stability around the target pH, assess the feasibility and responsibility associated with switching to PFS, and demonstrate good DP robustness against physicochemical stress.

[0320] PS80 concentration was evaluated by subjecting formulations containing different PS80 concentrations to mechanical stress. 0.06% (w / v) PS80 was identified as the surfactant concentration to mitigate the potential impact of PS80 loss on quality attributes.

[0321] Metal chelating agents were added to the formulation to provide protection from the degradation of potential metal-induced proteins and excipients. 10 μM EDTA and 10 μM DTPA provided comparable levels of protection. As a result, EDTA was ultimately selected.

[0322] The stability of amriterimab was evaluated in histidine buffer. DP stability studies in vials and PFS were conducted under refrigerated, ambient, accelerated, and frozen conditions between pH 5.5 and 6.3, demonstrating a favorable product profile. A 10 mM histidine buffer with a target pH of 6.0 was selected as the preferred buffer.

[0323] Sucrose is a well-proven excipient used in protein formulations as a stabilizer / freeze protector. 220 mM sucrose is approximately 7.5%, which represents a suitable ratio for protecting both DP and FDS concentrations between 31 and 125 mg / mL.

[0324] Furthermore, the stability of amriterimab was compared in both BD and OMPI PFS, demonstrating that they were similar except for the formation of fewer microscopic particles in BD PFS. DP stability studies in vials and PFS were conducted under refrigerated, ambient, accelerated, and frozen conditions between pH 5.5 and 6.3, demonstrating a favorable product profile.

[0325] The results of these stability studies comprehensively demonstrate the robustness of the targeted formulation for stabilizing aqueous amriterimab in frozen FDS and PFS under various storage and other significant stress conditions. Therefore, the final targeted amriterimab aqueous formulation derived from the results of this study, with pH 6.0+0.2, containing 10 mM histidine, 220 mM sucrose, 10 μM EDTA, and 0.06% PS80, is optimal for stabilizing DP at 31–125 mg / mL and FDS at 125 mg / mL.

[0326] Example drug This study led to the development of an exemplary amritelimab drug (DP). Amritelimab DP consists of amritelimab, L-histidine, L-histidine hydrochloride monohydrate, sucrose, EDTA, and ultra-purified polysorbate 80. The excipients in the amritelimab formulation are known to be well-tolerated after parenteral administration and to be water-soluble. The DP formulation and exemplary containers are summarized in Tables 1-7 below. [Table 9]

[0327] Example 2 - Product Quality of the Formulation Formulations F2-1, F2-2, and F2-3, as well as containers containing these formulations, were prepared as listed in Table 2-1. Formulation F2-1 (also referred herein as “Cycle 1” formulation) is distinguished from formulations F2-2 (also referred herein as “Cycle 2 ESB0.A_F1'”) and F2-3 (also referred herein as “Cycle 2” ESB0.B_F2'”) developed from the Cycle 2 study described in Example 1. These formulations were aseptically filled into 6 ml infusion vials (6R), sealed with FLUROTEC-coated stoppers and crimped with caps, or aseptically filled into BD Neopak using a 2.25 mL syringe closed with a West 1-3 mL Novapure 4023 / 50 grape plunger stopper. [Table 10]

[0328] Formulations F2-1, F2-2, and F2-3 were stored in containers at 5°C, 25°C, and 40°C for a maximum period of 3 months. The HMWS, PS80 degradation kinetics, deamidation, oxidation, and charge variants of the formulations were evaluated at T0, 1 month (1M), 2 months (2M), and 3 months (3M) according to the method detailed in Example 1. The results obtained are shown in Figures 38 to 42.

[0329] Figure 38 is a graph showing the change in HMWS analyzed by SEC-HPLC for formulations F2-1, F2-2, and F2-3 over 3 months of storage at 5°C, 25°C, and 40°C. As can be seen in Figure 38, the HMWS in formulations F2-1, F2-2, and F2-3 stored at specific temperatures such as 5°C, 25°C, or 40°C were comparable, with the highest HMWS% for all formulations after storage at 40°C for 2 months.

[0330] Figure 39 is a graph showing the PS80 loss of formulations F2-1, F2-2, and F2-3 over 3 months of storage at 5°C, 25°C, and 40°C, analyzed by CAD according to the method described in Example 1. The dotted line in Figure 39 indicates method variability. The results show that PS80 is stable in all formulations. The changes in PS80 over 3 months of storage at 5°C, 25°C, and 40°C were within the range of analytical variability (below the dotted line). The changes in PS80 in the three formulations stored at specific temperatures were similar, but the highest loss of PS80 was observed in F2-1 (Cycle 1) after 3 months of storage at 25°C.

[0331] Figure 40 is a graph showing the weekly percentage change in HC D332 deamidation of antibodies for formulations F2-1, F2-2, and F2-3 over a one-month storage period at 40°C, as measured by peptide mapping. As can be seen from Figure 40, there were no significant differences among the percentage changes in D332 deamidation for all three formulations measured.

[0332] Figure 41 is a graph showing the weekly percentage change in HC M103 oxidation of antibodies F2-1, F2-2, and F2-3, as measured by peptide mapping, over a one-month storage period at 40°C. As can be seen from Figure 41, there were no significant differences in the percentage change in M103 oxidation across all three formulations measured.

[0333] Figure 42 is a graph showing charge variants measured by capillary isoelectric focusing (cIEF) for formulations F2-1, F2-2, and F2-3 over two months of storage at 5°C, 25°C, and 40°C. As can be seen in Figure 43, the percentage of the major peak remained stable in all three formulations stored at 5°C and 25°C for up to three months. A slight decrease in the major peak began when the formulations were stored at 40°C for two weeks, and the percentage of the major peak remained decreasing over longer storage times at 40°C, with the lowest amount of major peak in the formulations stored at 40°C for two months. There were no significant differences in the major peaks among the formulations stored over time at the same temperature.

[0334] In summary, there were no significant differences in product quality, including physical or chemical properties or characteristics, between formulations F6-1, F6-2, and F6-3.

[0335] Examples 3 - The 28 mg / mL and 138 mg / mL formulations are stable. Formulations F3-1, F3-2, and F3-3 were prepared according to Table 3-1. Formulation F3-1 (also referred herein as “Cycle 1” formulation) was developed from an alternative study (Cycle 1 study) to develop formulations for Phase I and Phase II clinical trials. Formulations F3-2 (also referred herein as “Cycle 2 ESB0.B_1”) and F3-3 (also referred herein as “Cycle 2 ESB0.B_2”) were developed from the study described in Example 1. These formulations were aseptically filled into 6 mL injection vials (6R), sealed with FLUROTEC®-coated stoppers and crimped with caps, or aseptically filled into BD Neopak using a 2.25 mL syringe closed with a West 1-3 mL NOVAPURE® 4023 / 50 grape plunger stopper. [Table 11]

[0336] Formulations F3-1, F3-2, and F3-3 were stored in containers at 5°C, 25°C, and 40°C for a maximum period of 3 months. General product attributes, including visible particles and concentration, PS80 degradation kinetics, physical stability measured by HMWS and HIAC, purity by CE-SDA(NR), and chemical stability measured by charge variants by cIEF, deamidation, and oxidation, were evaluated over the 3-month storage period at 5°C, 25°C, and 40°C. The results are shown in Table 3-2 in Figure 43.

[0337] The results indicate that formulations F3-2 and F3-3 were stable over storage at 5°C, 25°C, and 40°C for at least two months. The results demonstrate that formulations containing specific concentrations of excipients developed in the study described in Example 1 were suitable for use with antibodies over a wide range of concentrations, from low to high. The formulations developed in Example 1 are suitable for producing pharmaceuticals with low to high concentrations of anti-hOX40L antibody for clinical or commercial purposes.

[0338] Example 4 - The 125 mg / mL PFS contained in BD and OMPI is stable. In this example, amriterimab was formulated in a solution containing 10 mM His, 220 mM sucrose, 0.04% PS80, and 10 μM EDTA in water at pH 6.0. The formulations were filled into 2.25 mL BD syringes and OMPI syringes, respectively. The products were stored at 40°C for at least one month. After storage at 40°C for one month, the gliding force of the product was evaluated. The results are shown in Figure 44. The gliding force results for 2.25 mL BD and OMPI PFS indicated that the product was stable over storage at 5°C for at least one year.

[0339] Example 5: Rheological characterization and viscosity screening for the development of anti-hOX40L antibody formulations. For rheology characterization, the DS of amritermab was concentrated upwards by centrifugation in 25 mM His / HisHCl buffer at pH 6.2 up to the maximum possible concentration (about 263 mg / ml). The concentration values of the product obtained at the start of the study were used for further dilution calculations required for the study.

[0340] Viscosity measurements were performed using an Anton Paar Rheometer (Graz, Austria). The viscoelastic behavior was studied at 25 °C with a shear rate ramp from 0 to 4000 s -1 in 25 mM histidine / histidine hydrochloride buffer (HisHCl) at pH 6.2 with a high concentration of 263 mg / ml of the antibody as shown in Figure 45. Amritermab showed a thinning of the shear force with increasing shear rate and, based on the thinning profile of the shear force, 2000 s -1 was selected as the shear force for all future studies.

[0341] Serial dilutions to five lower concentrations were performed using an Agilent Cary 60 spectrophotometer (Agilent Technologies, Santa Clara, CA, USA) with a variable pathlength extender (Repligen CTech™ SoloVPE®, Waltham, MA, USA) and verified by UV absorbance measurement (A280). The viscosity at each concentration in 25 mM histidine / histidine hydrochloride buffer (HisHCl) at pH 6.2 was measured at 25 °C and 5 °C at a shear rate of 2000 s -1 The concentration-dependent viscosity profile of amritermab obtained showed an exponential increase in viscosity at a concentration of about 150 mg / ml at 25 °C, but an exponential increase was observed at 5 °C after a concentration of about 100 mg / ml (Figure 46). The concentration and viscosity measurements obtained are described in more detail in Table 1a-1 below.

Table 12

[0342] Viscosity data at 25°C suggests that processing and increasing concentration can be achieved up to 150 mg / ml at room temperature under standard processing conditions. The target concentration for Part B was set at 180 mg / ml.

[0343] For viscosity reduction studies, amriterimab DS was subjected to buffer exchange in 20 mM His / HisHCl buffer at pH 6.0. After buffer exchange, the concentration was increased to approximately 230 mg / ml, followed by pH adjustment. Protein concentration and pH were monitored during the process. Stock solutions with different excipients were prepared to achieve the target formulation.

[0344] As shown in Table 1a-2, 12 formulation conditions with different excipients and pH levels were prepared for viscosity evaluation. Before viscosity measurement, all formulations were stored at 5°C for at least 1 day to assess for signs of visible phase transition. The viscosity of the different formulations was measured at 25°C for 2000 seconds. -1 The measurement was taken using the shear rate. The measurement was performed in duplicate. [Table 13]

[0345] The rheological property evaluations and viscosity reduction studies that were conducted are shown below. • Amriterimab treatment and elevation concentration in 20 mM HisHCl were successfully performed at different pH values. • The upward concentration process was faster at pH 5.0 and pH 5.5. A slower increase in concentration was observed in 20 mM HisHCl at pH 6.5. • At pH 5.0, pH 5.5, and pH 6.0, the target concentration of 180 mg / ml was successfully achieved by centrifugation in 20 mM HisHCl. The maximum concentration after compounding was 125 mg / mL for the formulation in 20 mM HisHCl at pH 6.5. The formulation containing 150 mM arginine-HCl showed a significant decrease in viscosity compared to the histidine-only formulation (approximately -60% depending on the pH). Furthermore, formulations containing 150 mM arginine-HCl were more effective in reducing viscosity compared to formulations containing 50 mM arginine-HCl. Preparations containing 50 mM or 150 mM sodium chloride showed a similar decrease in viscosity compared to histidine-only preparations (approximately -50% compared to histidine-only preparations at pH 6.0). The formulation containing 150 mM arginine-HCl showed slightly better viscosity reduction compared to 150 mM sodium chloride. The lowest viscosity measured for formulations containing 20 mM HisHCl and 150 mM arginine-HCl. • Sucrose increased viscosity compared to the histidine-only formulation (approximately +20% compared to the histidine-only formulation).

[0346] Based on the results obtained, it has been shown that arginine hydrochloride and sodium chloride effectively reduced the viscosity of amriterimab. Depending on pH, temperature, and formulation, a viscosity of approximately 15–25 mPas is expected for formulations without viscosity reducers at a target concentration of 125 mg / ml. In the presence of arginine hydrochloride or sodium chloride, a reduction to below 15 mPas may be expected. A target formulation concentration of 125 mg / mL at pH 6.0 is suggested for sc formulation development.

[0347] Example 6 - Development of an anti-OX40L antibody preparation The objective of this study was to screen pH, buffer, excipient, and surfactant concentrations under specific stress conditions to develop formulations containing a highly concentrated drug (DP) in aqueous form, i.e., liquid form, for subcutaneous (SC) administration of amriterimab.

[0348] Based on the research data from Example 5, different aqueous antibody formulations of anti-OX40L at a concentration of 125 mg / mL were prepared and investigated. An additional formulation at a relatively lower concentration of 75 mg / mL without viscosity-reducing excipients was also investigated. The aqueous formulations are listed in Table 2a-1. [Table 14]

[0349] The formulations listed in Table 2a-1 were prepared by 1) buffer exchange to achieve the target buffer concentration and pH, and then 2) increased concentration exceeding the target concentration was achieved.

[0350] Protein concentration, pH value, and density were determined during BPDS processing and used for the necessary calculations for each batch formulation using standard dilution procedures. The formulations were compounded by spiking them with the target formulation of the bulk drug along with the excipient stock solution. The protein concentration, osmotic pressure, and pH of the final compounded solution were determined and verified. All bulk drug (BDP) formulation solutions were filtered using a 0.22 μm polyvinylidene fluoride (PVDF) membrane filter.

[0351] Each formulation was manually filled into a 6R / 20mm glass type I vial, which was sealed with a 20mm bromobutyl rubber stopper and a 20mm aluminum flip-off seal, while observing sterile techniques.

[0352] Next, the prepared formulations were subjected to stability studies by placing them under thermal stress (up to 8 weeks for all formulations, and up to 16 weeks for F2a-1 and F2a-4, at -65°C or below, +5°C±3°C, and +25°C±2°C) and mechanical stress conditions (freezing / thawing and stirring). Each formulation sample was tested according to the test methods listed in Tables 2a-2. [Table 15]

[0353] The pH, protein concentration, and osmotic pressure values ​​obtained for the aqueous formulations (F2a-1, F2a-2, F2a-3, and F2a-4) after compounding and filtration were within the target range, as shown in Table 2a-3. [Table 16]

[0354] Data from stability studies at the initial time point T0 and after exposure to thermal stress, as well as data after freeze-thaw stress (-65°C to 25°C) or shaking stress at ambient temperature and low temperatures, are graphically shown in Figures 47 to 53C. Figure 47 is a graph showing the protein content of the formulation sample measured by UV / Vis. Figure 48 is a graph showing the pH value of the formulation sample. Figure 49 is a graph showing the surfactant content of the formulation sample. Figure 50 is a graph showing the clarity and opacity (turbidity) of the formulation sample solution. Figures 51A, 51B, and 51C are graphs showing the proportions of major peaks, HMW species, and LMW species obtained from SE-HPLC analysis of the formulation sample. Figure 52A graphically shows the count of invisible particles with a size of at least 2 μm for all formulation samples, with panel (a) showing counts in the range of 0 to 12000 (counts / mL) and panel (b) showing counts in the range of 0 to 2000 (counts / mL). Figure 52B graphically shows the count of invisible particles with a size of at least 5 μm (panel a), at least 10 (panel b), and at least 25 (panel (c)) for all formulation samples. Figures 53A, 53B, and 53C graphically show the major charge variant (major peak), acidic variant, and basic variant of the protein measured by icIEF.

[0355] Freeze-thaw and shaking studies As can be seen in Figures 47 to 50, all formulations subjected to freeze-thaw stress (-65°C to 25°C) or shaking stress at ambient temperature and low temperatures showed no significant changes in any of the analytical methods compared to the initial unstressed samples. This indicates that the formulations effectively stabilized the amriterimab molecule against both freeze-thaw and shaking stress.

[0356] Size exclusion chromatography (SE-HPLC) did not reveal significant differences in aggregation and fragmentation. Overall, the number of visible and invisible particles was low under shaking and freeze-thaw stress. All aqueous formulations were substantially free of visible particles under shaking and freeze-thaw stress. The data showed that both surfactants, polysorbate 20 and 80, protected the formulations from shaking, freeze, and thaw stress at the tested concentration levels (0.04% w / v at 125 mg / mL and 0.02% at 75 mg / mL).

[0357] Short-term stability study 1) Aggregation by SE-HPLC The short-term stability of formulation samples under different thermal stress conditions was analyzed for up to 8 weeks. Antibody purity was analyzed by size exclusion chromatography (SE-HPLC) to detect differences in the proportion of high molecular weight species (HMWS), major peaks, and low molecular weight species (LMWS) across all formulation samples.

[0358] The data showed that amriterimab molecules exhibited low to moderate aggregation tendencies at 5°C and 25°C, and more pronounced at 40°C. After 8 weeks of storage at 5°C, all formulations were comparable in area loss of up to 0.2% in major peak purity compared to T0 analyzed by SE-HPLC. After 8 weeks of storage at 25°C, an increase of less than 0.3% in area of ​​high molecular weight species (HMWS) was detected in all formulations. After 8 weeks at 40°C, an increase of 1.9% to 3.4% in HMWS was detected in all formulations. Formulations F2a-2 (containing NaCl) and F2a-3 (containing ArgHCl) showed similar degrees of HMWS formation, 3.4% and 3.3%, respectively, which were higher compared to formulations F2a-1 and F2a-4 at 125 mg / mL and 75 mg / mL, respectively. In particular, a larger increase in HMWS was observed, which was more pronounced compared to F2a-1 and F2a-4.

[0359] Further analysis of formulations F2a-1 and F2a-4 after 16 weeks of storage showed no significant changes at 5°C and 25°C. At 40°C, increases in the percentage of HMWS area of ​​6.3% and 5.2% compared to T0 were detected for formulations F2a-1 and F2a-4 at 125 mg / mL and 75 mg / mL, respectively. The change in major peak purity was minimally high in formulation F2a-1 at 8.9%, compared to an 8.2% area increase in F2a-4.

[0360] As can be seen in Figure 51C, after 8 weeks of storage at 40°C, all formulations showed a comparable increase in low molecular weight species (LMWS), with an area increase of 1.6% to 2.0% compared to T0. Slightly lower LMWS formation was detected for F2a-1 and F2a-4 compared to F2a-2 and F2a-3, which contained NaCl and ArgHCl as additional excipients, respectively. After 8 weeks of storage at 40°C, shoulders associated with LMWS could be separated for all formulations. Low fragmentation was observed at 2–8°C and 25°C. After 16 weeks of storage at 40°C, formulations F2a-1 and F2a-4 showed comparable increases in the percentage of LMWS area, of 2.6% and 2.9%, respectively.

[0361] A comparison of F2a-1 and F2a-4 at different protein concentrations after 16 weeks of storage revealed comparable aggregation and fragmentation behavior at 125 mg / mL and 75 mg / mL, respectively. Slightly higher HMWS formation was detected at higher protein concentrations, while LMWS formed independently of protein content. The change was that the formation of the primary peak purity, HMWS, and LMWS showed a non-linear trend dependent on protein concentration. The results suggest that aggregation and fragmentation rates did not change significantly when protein concentrations exceeded 75 mg / mL.

[0362] 2) Particles invisible to the naked eye As can be seen in Figures 52A and 52B, the overall initial invisible particle count was low to moderate for all formulations. In an 8-week stability study, formulations F2a-2 and F2a-3, containing NaCl and ArgHCl as additional excipients, respectively, showed an increase in invisible particle count, which was most pronounced at 40°C. After 16 weeks of storage, no significant change in invisible particle count was detected for formulations F2a-1 and F2a-4.

[0363] 3) Opaque light (turbidity) of the solution As can be seen in Figure 50, the opacity values ​​for formulations F2a-1 and F2a-4, with protein concentrations of 125 and 75 mg / ml, respectively, were approximately 15 NTU. On the other hand, the addition of NaCl (F2a-2) and ArgHCl (F2a-3) increased the turbidity to 30 NTU and 23 NTU, respectively. After storage at 40°C for 8 weeks, all formulations showed an increase in turbidity. No change in turbidity was observed for formulations stored at 2–8°C and 25°C.

[0364] 4) Charge variant Figures 53A, 53B, and 53C show the initial levels of charge variants as measured by iCIEF chromatograms. Moderate and major charge variants (major peaks) primarily become acidic variants at 25°C and 40°C after 8 and 16 weeks. All formulations showed comparable loss of major peak purity area (31.1%–34.4%) and formation of acidic variant area (31.4%–34.1%) after 8 weeks compared to T0. The data showed that the addition of NaCl or ArgHCl, or the reduction of protein concentration from 125 mg / mL to 75 mg / mL, did not affect the rate of acidic variant formation. After storage at 40°C for 16 weeks, formulations F2a-1 and F2a-4 showed changes of 57.4% and 58.6% in major peak purity, respectively, while the increase in acidic variants was 51.4% and 53.0%, respectively. iCIEF chromatograms revealed the transformation of major charge variants and basic variants into acidic variants.

[0365] 5) Purification and fragmentation by capillary electrophoresis Monomer purity was evaluated by tip-based CE-SDS and reverse-phase HPLC (RP-HPLC) under non-reducing conditions. The results from tip-based CE-SDS primarily showed loss of monomer peak purity at 40°C, as well as loss of total light and heavy chain purity (LC+HC). Heavy chain cleavage was mainly observed. However, overall fragmentation was moderate for formulations stored at other temperatures or subjected to shaking and freeze-thaw stress. Similar results were observed for reverse-phase HPLC (RP-HPLC) analysis, suggesting the presence of heavy chain cleavage primarily when the formulation was stored at 40°C, and were consistent across all formulations.

[0366] 6) Quantification of PS80 and / or PS20 Quantification of PS80 and PS20 showed a greater decrease in polysorbate (PS) 20 and 80 than in stability tests. PS20-containing formulations (F2a-2, F2a-3) showed comparable loss of surfactant content after 8 weeks at 25°C and 40°C. On the other hand, the loss of PS80 in F2a-1 and F2a-4 was lower at 25°C compared to PS20.

[0367] This study demonstrates that all aqueous formulations prepared and analyzed were stable against freeze-thaw and shaking stress (Figures 47–53C). Overall, amriterimab showed better stability in formulations without NaCl and ArgHCl after storage at 40°C for 8 weeks.

[0368] Formulations F2a-1 and F2a-4, which do not contain NaCl and ArgHCl, were further analyzed after 16 weeks of storage. After 16 weeks at 2–8°C (recommended storage temperature), no significant changes were observed in the physicochemical properties or major peak purity of formulation (F2a-1). The aqueous formulation (F2a-1) showed sufficient stability over 16 weeks at 25°C and exhibits sufficient stability under ambient conditions for handling during manufacturing. Temperature stress at 40°C induces increased physicochemical changes and chemical decomposition.

[0369] Based on the results from the study, the desired aqueous / liquid formulations, consistent with non-GMP and GMP pharmaceuticals, are recommended to contain 125 mg / mL amriterimab, 20 mM histidine / histidine HCl, 220 mM sucrose, and 0.04% (w / v) polysorbate 80, with a pH of approximately 6.0, and water.

[0370] Example 7 - Exemplary formulation and packaged product Exemplary aqueous pharmaceutical formulations F3a-1 and F3a-2 were prepared according to the formulations listed in Table 3a-1 below. [Table 17]

[0371] A container containing 1 mL of aqueous formulation F3a-1 or F3a-2 was developed. The container is a 2 mL Type 1 clear glass injection vial, in which aqueous formulation F3a-1 or F3a-2 was aseptically filled into such a vial with a nominal filling volume of 1 mL. The vial was then sealed with a FLUROTEC® coated stopper, and the stopper was crimped onto the vial neck with a cap to ensure the integrity of the container closure. For example, the stopper may be a 13 mm FLUROTEC® injection stopper manufactured by West Company, and the cap may be a 13 mm white FLIP-OFF® overcap manufactured by West design TruEdge. Formulations F3a-1 and F3a-2 in these glass vials are suitable for sc administration to subjects.

[0372] A pharmaceutical product containing 1 ml of aqueous formulation or solution of formulation F3a-1 or F3a-2 in each glass vial is suitable for use as a pharmaceutical unit dosage form. [Table 18-1] [Table 18-2] Table 18-3 Table 18-4 Table 18-5 Table 18-6 Table 18-7 Table 18-8 Table 18-9 Table 18-10 Table 18-11 Table 18-12 Table 18-13 Table 18-14 Table 18-15 Table 18-16 Table 18-17 Table 18-18 Table 18-19 [Table 18-20] [Table 18-21] [Table 18-22] [Table 18-23] [Table 18-24] [Table 18-25] [Table 18-26] [Table 18-27] [Table 18-28] [Table 18-29] [Table 18-30] [Table 18-31] [Table 18-32] IMGT indicates that the CDR is determined using the IMGT nomenclature. KABAT indicates that CDRs are determined using Kabat nomenclature. The numbering in the sequence correlation table takes precedence over any inconsistencies in numbering elsewhere in this specification.

[0373] This disclosure includes embodiments numbered as follows:

[0374] 1. (a) Anti-OX40 ligand (OX40L) antagonist antibody or its antigen-binding fragment, (b) Stabilizers and, (c) Chelating agents and, (d) Surfactants and (e) an aqueous pharmaceutical preparation comprising a buffer containing histidine or a histidine salt, The pH of the aqueous pharmaceutical preparation is in the range of 5.5 to approximately 6.5. The antibody or its antigen-binding fragment The heavy chain complementarity determination region (HCDR)1 of sequence number 42, HCDR2 of sequence number 44, and HCDR3 of sequence number 46, It includes the light chain complementarity determination region (LCDR) 1 of SEQ ID NO: 56, LCDR2 of SEQ ID NO: 58, and LCDR3 of SEQ ID NO: 60, The aqueous pharmaceutical formulation is suitable for parenteral administration to mammals.

[0375] 2. (a) Anti-OX40 ligand (OX40L) antagonist antibody or its antigen-binding fragment, (b) Stabilizers and, (c) Chelating agents and, (d) Surfactants and (e) an aqueous pharmaceutical preparation comprising a buffer containing histidine or a histidine salt, The pH of the aqueous pharmaceutical preparation is in the range of 5.5 to approximately 6.5. The antibody or its antigen-binding fragment Heavy chain complementarity determination region (HCDR)1 of SEQ ID NO: 36, HCDR2 of SEQ ID NO: 38, and HCDR3 of SEQ ID NO: 40, It includes the light chain complementarity determination region (LCDR) 1 of SEQ ID NO: 50, LCDR2 of SEQ ID NO: 52, and LCDR3 of SEQ ID NO: 54, The aqueous pharmaceutical formulation is suitable for parenteral administration to mammals.

[0376] 3. (a) Anti-OX40 ligand (OX40L) antagonist antibody or its antigen-binding fragment, (b) Stabilizers and, (c) Chelating agents and, (d) Surfactants and (e) an aqueous pharmaceutical preparation comprising a buffer containing histidine or a histidine salt, The pH of the aqueous pharmaceutical preparation is in the range of 5.5 to approximately 6.5. The antibody or its antigen-binding fragment comprises a VH domain having at least 80% identity with the amino acid sequence of SEQ ID NO: 34 and a VL domain having at least 80% identity with the amino acid sequence of SEQ ID NO: 48. The aqueous pharmaceutical formulation is suitable for parenteral administration to mammals.

[0377] 4. The aqueous pharmaceutical formulation according to Embodiment 1 or 2, wherein the antibody or antigen-binding fragment comprises a VH domain having at least 80% identity with the amino acid sequence of SEQ ID NO: 34 and / or a VL domain having at least 80% identity with the amino acid sequence of SEQ ID NO: 48.

[0378] 5. The aqueous pharmaceutical formulation according to any one of Embodiments 1 to 4, wherein the antibody or antigen-binding fragment comprises the VH domain of SEQ ID NO: 34 and / or the VL domain of SEQ ID NO: 48.

[0379] 6. An aqueous pharmaceutical preparation according to any one of Embodiments 1 to 5, wherein the antibody or antigen-binding fragment comprises a heavy chain and / or a light chain, the amino acid sequence of the heavy chain consists of the amino acid sequence of SEQ ID NO: 62, and / or the amino acid sequence of the light chain consists of the amino acid sequence of SEQ ID NO: 64.

[0380] 7. The aqueous pharmaceutical preparation according to any one of Embodiments 1 to 6, wherein the antibody or its antigen-binding fragment is amriterimab or a variant thereof.

[0381] 8. An aqueous pharmaceutical preparation according to any one of Embodiments 1 to 7, wherein the antibody or its antigen-binding fragment is present at a concentration in the range of 28 to 138 mg / mL or 31 to 125 mg / mL.

[0382] 9. An aqueous pharmaceutical preparation according to any one of Embodiments 1 to 8, wherein the antibody or its antigen-binding fragment is present at a concentration of 28 mg / mL, 31 mg / mL, 62.5 mg / mL, 125 mg / mL, or 138 mg / mL.

[0383] 10. An aqueous pharmaceutical preparation according to any one of Embodiments 1 to 9, wherein the antibody or its antigen-binding fragment is present at a concentration of about 62.5 mg / mL, preferably 62.5 mg / mL.

[0384] 11. An aqueous pharmaceutical preparation according to any one of Embodiments 1 to 9, wherein the antibody or its antigen-binding fragment is present at a concentration of about 125 mg / mL, preferably 125 mg / mL.

[0385] 12. The aqueous pharmaceutical preparation according to any one of Embodiments 1 to 11, wherein the stabilizer is sucrose.

[0386] 13. An aqueous pharmaceutical preparation according to any one of Embodiments 1 to 12, wherein at least one stabilizer is sucrose present in an amount of 220 mM.

[0387] 14. The aqueous pharmaceutical formulation according to any one of Embodiments 1 to 13, wherein the surfactant is polysorbate 80.

[0388] An aqueous pharmaceutical preparation according to any one of Embodiments 1 to 14, comprising 15.0.02% (w / v) to 0.1% (w / v) of polysorbate 80.

[0389] An aqueous pharmaceutical formulation according to any one of Embodiments 1 to 15, comprising 16.0.02% (w / v) to 0.06% polysorbate 80.

[0390] An aqueous pharmaceutical formulation according to any one of Embodiments 1 to 16, comprising 17.0.04% (w / v) or 0.06% polysorbate 80.

[0391] 18. The aqueous pharmaceutical preparation according to any one of Embodiments 1 to 17, wherein the buffer solution contains 10 mM L-histidine or histidine hydrochloride.

[0392] 19. An aqueous pharmaceutical formulation according to any one of Embodiments 1 to 18, further comprising a chelating agent selected from the group consisting of ethylenediaminetetraacetic acid (EDTA), diethylenetrimenpentaacetic acid (DTPA) and salts thereof, and any combination thereof.

[0393] 20. The aqueous pharmaceutical formulation according to Embodiment 19, wherein the chelating agent comprises one or both of 10 μM EDTA and 10 μM DTPA.

[0394] 21. The aqueous pharmaceutical formulation according to Embodiment 19, wherein the chelating agent contains 10 μM of EDTA.

[0395] The antibody or its antigen-binding fragment in a concentration of 22.28 mg / mL to 138 mg / mL, 10 mM ± 1.5 mM L-histidine or histidine hydrochloride, Sucrose of 220 mM ± 33 mM, 0.06% (w / v) polysorbate 80, Water and, The aqueous pharmaceutical preparation according to any one of Embodiments 1 to 7, wherein the pH of the aqueous pharmaceutical preparation is approximately 5.8 to approximately 6.2.

[0396] 23. The aqueous pharmaceutical formulation according to Embodiment 22, wherein the antibody or its antigen-binding fragment is present at a concentration of approximately 62.5 mg / mL, preferably 62.5 mg / mL.

[0397] 24. The aqueous pharmaceutical formulation according to Embodiment 22, wherein the antibody or its antigen-binding fragment is present at a concentration of about 125 mg / mL, preferably 125 mg / mL.

[0398] An aqueous pharmaceutical formulation according to any one of embodiments 22 to 24, further comprising 25.10 μM of EDTA or 10 μM of DTPA.

[0399] 26. The aqueous pharmaceutical formulation according to any one of Embodiments 22 to 25, wherein the aqueous pharmaceutical formulation does not contain a chelating agent or is essentially free of a chelating agent.

[0400] 27. An aqueous pharmaceutical formulation according to any one of Embodiments 1 to 26, which is free of or essentially free of particles.

[0401] 28. An aqueous pharmaceutical formulation according to any one of Embodiments 1 to 27, which does not contain or substantially contains sodium chloride.

[0402] 29. An aqueous pharmaceutical preparation according to any one of Embodiments 1 to 28, which does not contain or substantially contains arginine.

[0403] 30. An aqueous pharmaceutical formulation according to any one of Embodiments 1 to 29, wherein less than 5% of the antibody is detected in an aggregated form after storage at 40°C for 28 days and detected by size exclusion high-performance liquid chromatography.

[0404] An aqueous pharmaceutical preparation according to any one of Embodiments 1 to 30, which is stable when stored at 31.5°C or 25°C for at least 3 months.

[0405] An aqueous pharmaceutical preparation according to any one of Embodiments 1 to 31, which is stable when stored at 32.40°C for at least two months.

[0406] 33. An aqueous pharmaceutical preparation according to any one of Embodiments 1 to 32, which is stable for at least one year when stored at a temperature of approximately 5°C.

[0407] 34. An aqueous pharmaceutical preparation contained in a container, according to any one of Embodiments 1 to 33.

[0408] 35. An aqueous pharmaceutical formulation according to any one of Embodiments 1 to 34, which is suitable for subcutaneous delivery.

[0409] 36. A pharmaceutical unit dosage form suitable for parenteral administration to mammals, comprising an aqueous pharmaceutical preparation described in any one of Embodiments 1 to 35 in a suitable container.

[0410] 37. The pharmaceutical unit dosage form according to Embodiment 36, wherein the aqueous pharmaceutical preparation is suitable for intravenous, subcutaneous, or intramuscular administration.

[0411] 38. The pharmaceutical unit dosage form according to Embodiment 36 or 37, wherein the preferred container is a pre-filled syringe.

[0412] 39. A sealed container containing an aqueous pharmaceutical formulation according to any one of Embodiments 1 to 35.

[0413] 40. A sealed container according to Embodiment 39, which is a vial, syringe, microinfuser, pen delivery device, or auto-injector.

[0414] 41. A sealed container according to Embodiment 39 or 40, which is a single or multi-chamber syringe.

[0415] A sealed container according to Embodiment 41, which is a pre-filled syringe containing 42.2.25 mL of the aqueous pharmaceutical preparation.

[0416] 43. An anti-OX40 ligand (OX40L) antagonist antibody or its antigen-binding fragment in a concentration of approximately 28 mg / mL to approximately 138 mg / mL, Approximately 10 mM L-histidine or histidine hydrochloride, Approximately 220 mM sucrose, Approximately 0.06% (w / v) of polysorbate 80, A pre-filled syringe containing water and an aqueous pharmaceutical preparation, A pre-filled syringe containing the aqueous pharmaceutical preparation, with a pH of approximately 6.0.

[0417] 44. An anti-OX40 ligand (OX40L) antagonist antibody or its antigen-binding fragment in a concentration of approximately 28 mg / mL to approximately 138 mg / mL, Approximately 10 mM L-histidine or histidine hydrochloride, Approximately 220 mM sucrose, Approximately 0.06% (w / v) of polysorbate 80, A pre-filled pen or auto-injector containing water and an aqueous pharmaceutical preparation, A pre-filled pen or auto-injector in which the pH of the aqueous pharmaceutical preparation is approximately 6.0.

[0418] A pre-filled syringe according to Embodiment 43, or a pre-filled pen or auto-injector according to Embodiment 44, further comprising 45.10 μM EDTA or 10 μM DTPA.

[0419] A pre-filled syringe according to Embodiment 43, or a pre-filled pen or auto-injector according to Embodiment 44, further comprising 46.10 μM EDTA.

[0420] 47. A pre-filled syringe according to any one of Embodiments 43, 45, or 46, or a pre-filled pen or auto-injector according to any one of Embodiments 44 to 46, wherein the antibody or its antigen-binding fragment is present at a concentration of approximately 62.5 mg / mL, preferably 62.5 mg / mL.

[0421] 48. A pre-filled syringe according to any one of Embodiments 43, 45, or 46, or a pre-filled pen or auto-injector according to any one of Embodiments 44 to 46, wherein the antibody or its antigen-binding fragment is present at a concentration of about 125 mg / mL, preferably 125 mg / mL.

[0422] 49. The anti-OX40 ligand (OX40L) antagonist antibody or its antigen-binding fragment (a) Heavy chain complementarity determination region (HCDR) 1 of SEQ ID NO: 42, HCDR2 of SEQ ID NO: 44, and HCDR3 of SEQ ID NO: 46, and Light chain complementarity determination region (LCDR) 1 of SEQ ID NO: 56, LCDR2 of SEQ ID NO: 58, and LCDR3 of SEQ ID NO: 60, or (b) Heavy chain complementarity determination region (HCDR) 1 of SEQ ID NO: 36, HCDR2 of SEQ ID NO: 38, and HCDR3 of SEQ ID NO: 40, and Light chain complementarity determination region (LCDR) 1 of SEQ ID NO: 50, LCDR2 of SEQ ID NO: 52, and LCDR3 of SEQ ID NO: 54, or (c) A VH domain having at least 80% identity with the amino acid sequence of SEQ ID NO: 34, and a VL domain having at least 80% identity with the amino acid sequence of SEQ ID NO: 48, or (d) A VH domain containing a VL domain that has at least 80% identity to the amino acid sequence of SEQ ID NO: 34 and / or has at least 80% identity to the amino acid sequence of SEQ ID NO: 48, or (e) A pre-filled syringe according to any one of Embodiments 43 or 45-48, comprising a heavy chain amino acid sequence consisting of the amino acid sequence of Sequence ID No. 62 and / or a light chain amino acid sequence consisting of the amino acid sequence of Sequence ID No. 64, or a pre-filled pen or auto-injector according to any one of Embodiments 44-48.

[0423] 50. A pre-filled syringe according to Embodiment 43 or any one of Embodiments 45-49, or a pre-filled pen or auto-injector according to any one of Embodiments 44-49, containing amliterimab or a variant thereof in a concentration of approximately 31 mg / mL to approximately 125 mg / mL.

[0424] A pre-filled syringe according to any one of Embodiments 43 or 45-50, or a pre-filled pen or auto-injector according to any one of Embodiments 44-50, containing 51.62.5 mg / mL of amliterimab or a variant thereof.

[0425] A pre-filled syringe according to any one of Embodiments 43 or 45-50, or a pre-filled pen or auto-injector according to any one of Embodiments 44-50, containing 52.125 mg / mL of amriterimab or a variant thereof.

[0426] 53. A kit comprising a sealed container according to any one of embodiments 39 to 42, a pre-filled syringe according to any one of embodiments 43 or 45 to 52, or a pre-filled pen or auto-injector according to any one of embodiments 44 to 52.

[0427] 54. A sealed container containing the aqueous pharmaceutical formulation described in any one of Embodiments 1 to 35, A kit comprising the aqueous pharmaceutical formulation and at least one separate injection device for delivering it to a mammalian subject in need.

[0428] 55. The kit according to Embodiment 54, wherein the injection device is a syringe, a microinfuser, a pen delivery device, or an auto-injector.

[0429] 56. The kit according to embodiment 55, wherein the injection device is a single or multi-chamber syringe.

[0430] 57. For use in the treatment of a disease or condition in a subject selected from the group consisting of autoimmune diseases or conditions, inflammatory diseases or conditions, systemic inflammatory diseases or conditions, and transplant rejection diseases or conditions, mediated by hOX40L, an aqueous pharmaceutical formulation according to any one of Embodiments 1 to 35, a pharmaceutical unit dosage form according to any one of Embodiments 36 to 38, a sealed container according to any one of Embodiments 39 to 42, a pre-filled syringe according to any one of Embodiments 43 or 45 to 52, a pre-filled pen or auto-injector according to any one of Embodiments 44 to 52, or a kit according to any one of Embodiments 53 to 56.

[0431] 58. An aqueous pharmaceutical formulation according to any one of Embodiments 1 to 35, a pharmaceutical unit dosage form according to any one of Embodiments 36 to 38, a sealed container according to any one of Embodiments 39 to 42, a pre-filled syringe according to any one of Embodiments 43 or 45 to 52, a pre-filled pen or auto-injector according to any one of Embodiments 44 to 52, or a kit according to any one of Embodiments 53 to 56, for use in the treatment of atopic dermatitis.

[0432] 59. An aqueous pharmaceutical formulation according to any one of Embodiments 1 to 35, a pharmaceutical unit dosage form according to any one of Embodiments 36 to 38, a sealed container according to any one of Embodiments 39 to 42, a pre-filled syringe according to any one of Embodiments 43 or 45 to 52, a pre-filled pen or auto-injector according to any one of Embodiments 44 to 52, or a kit according to any one of Embodiments 53 to 56, for use in the treatment of asthma.

[0433] A compound for the manufacture of a pharmacopoeia for the treatment of a disease or condition in a subject selected from the group consisting of autoimmune diseases or conditions, inflammatory diseases or conditions, systemic inflammatory diseases or conditions, and transplant rejection diseases or conditions, mediated by 60.hOX40L, comprising an aqueous pharmaceutical formulation according to any one of Embodiments 1 to 35, a pharmaceutical unit dosage form according to any one of Embodiments 36 to 38, a sealed container according to any one of Embodiments 39 to 42, a pre-filled syringe according to any one of Embodiments 43 or 45 to 52, a pre-filled pen or auto-injector according to any one of Embodiments 44 to 52, or a kit according to any one of Embodiments 53 to 56.

[0434] 61. In the manufacture of a pharmaceutical product for the treatment of atopic dermatitis, an aqueous pharmaceutical product according to any one of Embodiments 1 to 35, a pharmaceutical unit dosage form according to any one of Embodiments 36 to 38, a sealed container according to any one of Embodiments 39 to 42, a pre-filled syringe according to any one of Embodiments 43 or 45 to 52, a pre-filled pen or auto-injector according to any one of Embodiments 44 to 52, or a kit according to any one of Embodiments 53 to 56.

[0435] 62. An aqueous pharmaceutical formulation according to any one of Embodiments 1 to 35, a pharmaceutical unit dosage form according to any one of Embodiments 36 to 38, a sealed container according to any one of Embodiments 39 to 42, a pre-filled syringe according to any one of Embodiments 43 or 45 to 52, a pre-filled pen or auto-injector according to any one of Embodiments 44 to 52, or a kit according to any one of Embodiments 53 to 56.

[0436] 63. A method for treating a disease or condition in a subject selected from the group consisting of autoimmune diseases or conditions, inflammatory diseases or conditions, systemic inflammatory diseases or conditions, and transplant rejection diseases or conditions (e.g., autoimmune diseases or conditions, inflammatory diseases or conditions, systemic inflammatory diseases or conditions, and transplant rejection diseases or conditions mediated by hOX40L), comprising administering to the subject an effective amount of an aqueous pharmaceutical preparation described in any one of Embodiments 1 to 35.

[0437] 64. A method for treating atopic dermatitis, comprising administering to a subject an effective amount of an aqueous pharmaceutical preparation described in any one of Embodiments 1 to 35.

[0438] 65. A method for treating asthma, comprising administering to a subject an effective amount of an aqueous pharmaceutical preparation described in any one of Embodiments 1 to 35.

[0439] 66. (a) Anti-OX40 ligand (OX40L) antagonist antibody or antigen-binding fragment, (b) Stabilizers and, (c) Chelating agents and, (d) Surfactants and (e) an aqueous pharmaceutical preparation comprising a buffer containing histidine or a histidine salt, The pH of the aqueous pharmaceutical preparation is in the range of 5.5 to approximately 6.5. The antibody or its antigen-binding fragment A VH domain containing the HCDR1 sequence of sequence number 36 or 42, the HCDR2 sequence of sequence number 38 or 44, and the HCDR3 sequence of sequence number 40 or 46, A VL domain comprising the LCDR1 sequence of sequence number 50 or 56, the LCDR2 sequence of sequence number 52 or 58, and the LCDR3 sequence of sequence number 54 or 60, The aqueous pharmaceutical formulation is suitable for parenteral administration to mammals.

[0440] 67. The method according to Embodiment 66, wherein the VL domain comprises the amino acid sequence of SEQ ID NO: 48 and the VH domain comprises the amino acid sequence of SEQ ID NO: 34.

[0441] 68. The method according to Embodiment 66 or 67, wherein the antibody comprises a heavy chain and a light chain, the amino acid sequence of the heavy chain is the amino acid sequence of SEQ ID NO: 62, and the amino acid sequence of the light chain is the amino acid sequence of SEQ ID NO: 64.

[0442] 69. (a) Approximately 62.5 mg (e.g., 62.15 mg) of an anti-OX40 ligand (OX40L) antagonist antibody or its antigen-binding fragment, (b) Approximately 1.49 mg of L-histidine and (c) Approximately 2.18 mg of L-histidine hydrochloride (e.g., L-histidine hydrochloride monohydrate) (d) Approximately 150.62 mg of sucrose, (e) Approximately 0.0074 mg of EDTA (e.g., EDTA disodium dihydrate) and (f) Approximately 1.2 mg of polysorbate 80 (e.g., ultra-purified polysorbate 80) and (g) A pharmaceutical dosage form suitable for parenteral administration to mammalian subjects, optionally containing water, The antibody or its antigen-binding fragment A VH domain containing the HCDR1 sequence of sequence number 36 or 42, the HCDR2 sequence of sequence number 38 or 44, and the HCDR3 sequence of sequence number 40 or 46, A pharmaceutical dosage form comprising a VL domain containing the LCDR1 sequence of SEQ ID NO: 50 or 56, the LCDR2 sequence of SEQ ID NO: 52 or 58, and the LCDR3 sequence of SEQ ID NO: 54 or 60.

[0443] 70.(a) Approximately 125 mg of an anti-OX40 ligand (OX40L) antagonist antibody or its antigen-binding fragment, (b) Approximately 1.49 mg of L-histidine and (c) Approximately 2.18 mg of L-histidine hydrochloride (e.g., L-histidine hydrochloride monohydrate) (d) Approximately 150.62 mg of sucrose, (e) Approximately 0.0074 mg of EDTA (e.g., EDTA disodium dihydrate) and (f) Approximately 1.2 mg of ultra-purified polysorbate 80, (g) A pharmaceutical dosage form suitable for parenteral administration to mammalian subjects, optionally containing water, The antibody or its antigen-binding fragment A VH domain containing the HCDR1 sequence of sequence number 36 or 42, the HCDR2 sequence of sequence number 38 or 44, and the HCDR3 sequence of sequence number 40 or 46, A pharmaceutical dosage form comprising a VL domain containing the LCDR1 sequence of SEQ ID NO: 50 or 56, the LCDR2 sequence of SEQ ID NO: 52 or 58, and the LCDR3 sequence of SEQ ID NO: 54 or 60.

[0444] 71. (a) Approximately 250 mg of anti-OX40 ligand (OX40L) antagonist antibody or its antigen-binding fragment, (b) Approximately 1.49 mg of L-histidine and (c) Approximately 2.18 mg of L-histidine hydrochloride (e.g., L-histidine hydrochloride monohydrate) (d) Approximately 150.62 mg of sucrose, (e) Approximately 0.0074 mg of EDTA (e.g., EDTA disodium dihydrate) and (f) Approximately 1.2 mg of polysorbate 80 (e.g., ultra-purified polysorbate 80) and (g) A pharmaceutical dosage form suitable for parenteral administration to mammalian subjects, optionally containing water, The antibody or its antigen-binding fragment A VH domain containing the HCDR1 sequence of sequence number 36 or 42, the HCDR2 sequence of sequence number 38 or 44, and the HCDR3 sequence of sequence number 40 or 46, A pharmaceutical dosage form comprising a VL domain containing the LCDR1 sequence of SEQ ID NO: 50 or 56, the LCDR2 sequence of SEQ ID NO: 52 or 58, and the LCDR3 sequence of SEQ ID NO: 54 or 60.

[0445] 72. The pharmaceutical unit dosage form according to any one of Embodiments 69 to 71, wherein the anti-OX40 ligand (OX40L) antagonist antibody or its antigen-binding fragment is amriterimab or a variant thereof.

[0446] 73. A pharmaceutical unit dosage form according to any one of embodiments 69 to 71, wherein the unit dosage form is contained in a suitable container.

[0447] 74. The pharmaceutical unit dosage form according to Embodiment 73, wherein the preferred container is a vial, a syringe (e.g., a pre-filled syringe), a microinfuser, a pen delivery device (e.g., a pre-filled pen delivery device), or an auto-injector.

[0448] 75. The pharmaceutical unit dosage form according to embodiment 73 or 74, wherein the container is sealed.

[0449] This disclosure includes the following numbered aspects:

[0450] 1. (a) Anti-OX40 ligand (OX40L) antagonist antibody or its antigen-binding fragment, (b) Stabilizers and, (c) Chelating agents and, (d) Surfactants and (e) A pharmaceutical preparation comprising a buffer containing histidine or a histidine salt, The pH of the aforementioned pharmaceutical preparation is in the range of 5.5 to approximately 6.5. The antibody or its antigen-binding fragment The heavy chain complementarity determination region (HCDR)1 of sequence number 42, HCDR2 of sequence number 44, and HCDR3 of sequence number 46, It includes the light chain complementarity determination region (LCDR) 1 of SEQ ID NO: 56, LCDR2 of SEQ ID NO: 58, and LCDR3 of SEQ ID NO: 60, A pharmaceutical formulation which is suitable for parenteral administration to mammals.

[0451] 2. (a) Anti-OX40 ligand (OX40L) antagonist antibody or its antigen-binding fragment, (b) Stabilizers and, (c) Chelating agents and, (d) Surfactants and (e) A pharmaceutical preparation comprising a buffer containing histidine or a histidine salt, The pH of the aforementioned pharmaceutical preparation is in the range of 5.5 to approximately 6.5. The antibody or its antigen-binding fragment Heavy chain complementarity determination region (HCDR)1 of SEQ ID NO: 36, HCDR2 of SEQ ID NO: 38, and HCDR3 of SEQ ID NO: 40, It includes the light chain complementarity determination region (LCDR) 1 of SEQ ID NO: 50, LCDR2 of SEQ ID NO: 52, and LCDR3 of SEQ ID NO: 54, A pharmaceutical formulation which is suitable for parenteral administration to mammals.

[0452] 3. (a) Anti-OX40 ligand (OX40L) antagonist antibody or its antigen-binding fragment, (b) Stabilizers and, (c) Chelating agents and, (d) Surfactants and (e) A pharmaceutical preparation comprising a buffer containing histidine or a histidine salt, The pH of the aforementioned pharmaceutical preparation is in the range of 5.5 to approximately 6.5. The antibody or its antigen-binding fragment comprises a VH domain having at least 80% identity with the amino acid sequence of SEQ ID NO: 34 and a VL domain having at least 80% identity with the amino acid sequence of SEQ ID NO: 48. A pharmaceutical formulation which is suitable for parenteral administration to mammals.

[0453] 4. The pharmaceutical preparation according to embodiment 1 or 2, wherein the antibody or antigen-binding fragment comprises a VH domain having at least 80% identity with the amino acid sequence of SEQ ID NO: 34 and / or a VL domain having at least 80% identity with the amino acid sequence of SEQ ID NO: 48.

[0454] 5. A pharmaceutical preparation according to any one of embodiments 1 to 4, wherein the antibody or antigen-binding fragment comprises the VH domain of SEQ ID NO: 34 and / or the VL domain of SEQ ID NO: 48.

[0455] 6. A pharmaceutical preparation according to any one of embodiments 1 to 5, wherein the antibody or antigen-binding fragment comprises a heavy chain and / or a light chain, the amino acid sequence of the heavy chain consists of the amino acid sequence of SEQ ID NO: 62, and / or the amino acid sequence of the light chain consists of the amino acid sequence of SEQ ID NO: 64.

[0456] 7. A pharmaceutical preparation according to any one of embodiments 1 to 6, wherein the antibody or its antigen-binding fragment is amriterimab or a variant thereof.

[0457] 8. A pharmaceutical preparation according to any one of embodiments 1 to 7, wherein the antibody or its antigen-binding fragment is present at a concentration in the range of 28 to 138 mg / mL or 31 to 125 mg / mL.

[0458] 9. A pharmaceutical preparation according to any one of embodiments 1 to 8, wherein the antibody or its antigen-binding fragment is present at a concentration of 28 mg / mL, 31 mg / mL, 62.5 mg / mL, 125 mg / mL, or 138 mg / mL.

[0459] 10. A pharmaceutical preparation according to any one of embodiments 1 to 9, wherein the antibody or its antigen-binding fragment is present at a concentration of approximately 62.5 mg / mL, preferably 62.5 mg / mL.

[0460] 11. A pharmaceutical preparation according to any one of embodiments 1 to 9, wherein the antibody or its antigen-binding fragment is present at a concentration of about 125 mg / mL, preferably 125 mg / mL.

[0461] 12. A pharmaceutical preparation according to any one of embodiments 1 to 11, wherein the stabilizer is sucrose.

[0462] 13. A pharmaceutical preparation according to any one of embodiments 1 to 12, wherein at least one stabilizer is sucrose present in an amount of 220 mM.

[0463] 14. A pharmaceutical preparation according to any one of embodiments 1 to 13, wherein the surfactant is polysorbate 80.

[0464] A pharmaceutical preparation according to any one of embodiments 1 to 14, comprising 15.0.02% (w / v) to 0.1% (w / v) of polysorbate 80.

[0465] A pharmaceutical preparation according to any one of embodiments 1 to 15, comprising 16.0.02% (w / v) to 0.06% polysorbate 80.

[0466] A pharmaceutical preparation according to any one of embodiments 1 to 16, comprising 17.0.04% (w / v) or 0.06% polysorbate 80.

[0467] 18. A pharmaceutical preparation according to any one of embodiments 1 to 17, wherein the buffer solution contains 10 mM L-histidine or histidine hydrochloride.

[0468] 19. A pharmaceutical preparation according to any one of embodiments 1 to 18, further comprising a chelating agent selected from the group consisting of ethylenediaminetetraacetic acid (EDTA), diethylenetrimenpentaacetic acid (DTPA), salts thereof, and any combination thereof.

[0469] 20. The pharmaceutical formulation according to embodiment 19, wherein the chelating agent comprises one or both of 10 μM EDTA and 10 μM DTPA.

[0470] 21. The pharmaceutical preparation according to embodiment 19, wherein the chelating agent contains 10 μM of EDTA.

[0471] The antibody or its antigen-binding fragment in a concentration of 22.28 mg / mL to 138 mg / mL, 10 mM ± 1.5 mM L-histidine or histidine hydrochloride, Sucrose of 220 mM ± 33 mM, 0.06% (w / v) polysorbate 80, Water and, The pharmaceutical preparation according to any one of embodiments 1 to 7, wherein the pH of the aqueous pharmaceutical preparation is approximately 5.8 to approximately 6.2.

[0472] 23. The pharmaceutical preparation according to embodiment 22, wherein the antibody or its antigen-binding fragment is present at a concentration of approximately 62.5 mg / mL, preferably 62.5 mg / mL.

[0473] 24. The pharmaceutical preparation according to embodiment 22, wherein the antibody or its antigen-binding fragment is present at a concentration of about 125 mg / mL, preferably 125 mg / mL.

[0474] A pharmaceutical preparation according to any one of embodiments 22 to 24, further comprising 25.10 μM of EDTA or 10 μM of DTPA.

[0475] 26. The pharmaceutical preparation according to any one of embodiments 22 to 25, wherein the pharmaceutical preparation does not contain a chelating agent or is essentially free of a chelating agent.

[0476] 27. A pharmaceutical preparation according to any one of embodiments 1 to 26, which does not contain or is essentially free of particles.

[0477] 28. A pharmaceutical preparation according to any one of embodiments 1 to 27, which does not contain or substantially contains sodium chloride.

[0478] 29. A pharmaceutical preparation according to any one of embodiments 1 to 28, which does not contain or substantially contains arginine.

[0479] 30. A pharmaceutical preparation according to any one of embodiments 1 to 29, wherein less than 5% of the antibody is detected in an aggregated form after storage at 40°C for 28 days and detected by size exclusion high-performance liquid chromatography.

[0480] A pharmaceutical preparation according to any one of embodiments 1 to 30, which is stable when stored at 31.5°C or 25°C for at least 3 months.

[0481] A pharmaceutical preparation according to any one of embodiments 1 to 31, which is stable when stored at 32.40°C for at least two months.

[0482] 33. A pharmaceutical preparation according to any one of embodiments 1 to 32, which is stable for at least one year when stored at a temperature of approximately 5°C.

[0483] 34. A pharmaceutical preparation contained in a container, as described in any one of embodiments 1 to 33.

[0484] 35. A pharmaceutical preparation according to any one of embodiments 1 to 34, which is suitable for subcutaneous delivery.

[0485] 36. A pharmaceutical unit dosage form suitable for parenteral administration to mammals, comprising a pharmaceutical preparation described in any one of embodiments 1 to 35 in a suitable container.

[0486] 37. The pharmaceutical formulation according to embodiment 36, wherein the pharmaceutical formulation is suitable for intravenous, subcutaneous, or intramuscular administration.

[0487] 38. The pharmaceutical unit dosage form according to embodiment 36 or 37, wherein the preferred container is a pre-filled syringe.

[0488] 39. A sealed container containing a pharmaceutical preparation as described in any one of embodiments 1 to 35.

[0489] 40. A sealed container according to embodiment 39, which is a vial, syringe, microinfuser, pen delivery device, or auto-injector.

[0490] 41. A sealed container according to embodiment 39 or 40, which is a single or multi-chamber syringe.

[0491] A sealed container according to embodiment 41, which is a pre-filled syringe containing 42.2.25 mL of the pharmaceutical preparation.

[0492] 43. An anti-OX40 ligand (OX40L) antagonist antibody or its antigen-binding fragment in a concentration of approximately 28 mg / mL to approximately 138 mg / mL, Approximately 10 mM L-histidine or histidine hydrochloride, Approximately 220 mM sucrose, Approximately 0.06% (w / v) of polysorbate 80, A pre-filled syringe containing water and a pharmaceutical preparation, A pre-filled syringe containing the aforementioned pharmaceutical preparation, with a pH of approximately 6.0.

[0493] 44. An anti-OX40 ligand (OX40L) antagonist antibody or its antigen-binding fragment in a concentration of approximately 28 mg / mL to approximately 138 mg / mL, Approximately 10 mM L-histidine or histidine hydrochloride, Approximately 220 mM sucrose, Approximately 0.06% (w / v) of polysorbate 80, A pre-filled pen or auto-injector containing water and a pharmaceutical preparation, A pre-filled pen or auto-injector in which the pH of the aforementioned pharmaceutical preparation is approximately 6.0.

[0494] A pre-filled syringe according to embodiment 43, or a pre-filled pen or auto-injector according to embodiment 44, further comprising 45.10 μM EDTA or 10 μM DTPA.

[0495] A pre-filled syringe according to embodiment 43, or a pre-filled pen or auto-injector according to embodiment 44, further comprising 46.10 μM EDTA.

[0496] 47. A pre-filled syringe according to any one of embodiments 43, 45, or 46, or a pre-filled pen or auto-injector according to any one of embodiments 44 to 46, wherein the antibody or its antigen-binding fragment is present at a concentration of approximately 62.5 mg / mL, preferably 62.5 mg / mL.

[0497] 48. A pre-filled syringe according to any one of embodiments 43, 45, or 46, or a pre-filled pen or auto-injector according to any one of embodiments 44 to 46, wherein the antibody or its antigen-binding fragment is present at a concentration of about 125 mg / mL, preferably 125 mg / mL.

[0498] 49. The anti-OX40 ligand (OX40L) antagonist antibody or its antigen-binding fragment (a) Heavy chain complementarity determination region (HCDR) 1 of SEQ ID NO: 42, HCDR2 of SEQ ID NO: 44, and HCDR3 of SEQ ID NO: 46, and Light chain complementarity determination region (LCDR) 1 of SEQ ID NO: 56, LCDR2 of SEQ ID NO: 58, and LCDR3 of SEQ ID NO: 60, or (b) Heavy chain complementarity determination region (HCDR) 1 of SEQ ID NO: 36, HCDR2 of SEQ ID NO: 38, and HCDR3 of SEQ ID NO: 40, and Light chain complementarity determination region (LCDR) 1 of SEQ ID NO: 50, LCDR2 of SEQ ID NO: 52, and LCDR3 of SEQ ID NO: 54, or (c) A VH domain having at least 80% identity with the amino acid sequence of SEQ ID NO: 34, and a VL domain having at least 80% identity with the amino acid sequence of SEQ ID NO: 48, or (d) A VH domain containing a VL domain that has at least 80% identity to the amino acid sequence of SEQ ID NO: 34 and / or has at least 80% identity to the amino acid sequence of SEQ ID NO: 48, or ...

Claims

1. (a) an anti-OX40 ligand (OX40L) antagonist antibody or its antigen-binding fragment, (b) Stabilizers and, (c) Chelating agent, (d) Surfactants and (e) an aqueous pharmaceutical preparation comprising a buffer containing histidine or a histidine salt, The pH of the aqueous pharmaceutical preparation is in the range of 5.5 to approximately 6.

5. The antibody or its antigen-binding fragment The heavy chain complementarity determination region (HCDR) 1 of SEQ ID NO: 42, HCDR 2 of SEQ ID NO: 44, and HCDR 3 of SEQ ID NO: 46, An aqueous pharmaceutical formulation comprising the light chain complementarity determining region (LCDR) 1 of SEQ ID NO: 56, the LCDR 2 of SEQ ID NO: 58, and the LCDR 3 of SEQ ID NO:

60.

2. (a) an anti-OX40 ligand (OX40L) antagonist antibody or its antigen-binding fragment, (b) Stabilizers, (c) Chelating agent, (d) Surfactants and (e) an aqueous pharmaceutical preparation comprising a buffer containing histidine or a histidine salt, The pH of the aqueous pharmaceutical preparation is in the range of 5.5 to approximately 6.

5. The antibody or its antigen-binding fragment The heavy chain complementarity determination region (HCDR) 1 of SEQ ID NO: 36, HCDR 2 of SEQ ID NO: 38, and HCDR 3 of SEQ ID NO: 40, An aqueous pharmaceutical formulation comprising the light chain complementarity determining region (LCDR) 1 of SEQ ID NO: 50, the LCDR 2 of SEQ ID NO: 52, and the LCDR 3 of SEQ ID NO:

54.

3. (a) an anti-OX40 ligand (OX40L) antagonist antibody or its antigen-binding fragment, (b) Stabilizers, (c) Chelating agent, (d) Surfactants and (e) an aqueous pharmaceutical preparation comprising a buffer containing histidine or a histidine salt, The pH of the aqueous pharmaceutical preparation is in the range of 5.5 to approximately 6.

5. An aqueous pharmaceutical preparation wherein the antibody or its antigen-binding fragment comprises a VH domain having at least 80% identity with the amino acid sequence of SEQ ID NO: 34 and a VL domain having at least 80% identity with the amino acid sequence of SEQ ID NO:

48.

4. The aqueous pharmaceutical formulation according to claim 1 or 2, wherein the antibody or antigen-binding fragment comprises a VH domain having at least 80% identity with the amino acid sequence of SEQ ID NO: 34 and / or a VL domain having at least 80% identity with the amino acid sequence of SEQ ID NO:

48.

5. The aqueous pharmaceutical formulation according to any one of claims 1 to 4, wherein the antibody or antigen-binding fragment comprises the VH domain of SEQ ID NO: 34 and / or the VL domain of SEQ ID NO:

48.

6. The aqueous pharmaceutical preparation according to any one of claims 1 to 5, wherein the antibody or antigen-binding fragment comprises a heavy chain and / or a light chain, the amino acid sequence of the heavy chain consists of the amino acid sequence of SEQ ID NO: 62, and / or the amino acid sequence of the light chain consists of the amino acid sequence of SEQ ID NO:

64.

7. The aqueous pharmaceutical preparation according to any one of claims 1 to 6, wherein the antibody or its antigen-binding fragment is amriterimab or a variant thereof.

8. The aqueous pharmaceutical formulation according to any one of claims 1 to 7, wherein the aqueous pharmaceutical formulation is suitable for parenteral administration to mammals.

9. The aqueous pharmaceutical preparation according to any one of claims 1 to 8, wherein the antibody or its antigen-binding fragment is present at a concentration of 62.5 mg / mL.

10. The aqueous pharmaceutical preparation according to any one of claims 1 to 8, wherein the antibody or its antigen-binding fragment is present at a concentration of 125 mg / mL.

11. The aqueous pharmaceutical preparation according to any one of claims 1 to 10, wherein the stabilizer is sucrose.

12. The aqueous pharmaceutical preparation according to any one of claims 1 to 11, wherein at least one of the stabilizers is sucrose present in an amount of 220 mM.

13. The aqueous pharmaceutical preparation according to any one of claims 1 to 12, wherein the surfactant is polysorbate 80.

14. An aqueous pharmaceutical preparation according to any one of claims 1 to 13, comprising 0.02% (w / v) to 0.1% (w / v) of polysorbate 80.

15. An aqueous pharmaceutical preparation according to any one of claims 1 to 14, comprising 0.02% (w / v) to 0.06% polysorbate 80.

16. An aqueous pharmaceutical formulation according to any one of claims 1 to 15, comprising 0.04% (w / v) or 0.06% polysorbate 80.

17. The aqueous pharmaceutical preparation according to any one of claims 1 to 16, wherein the buffer solution contains 10 mM L-histidine or histidine hydrochloride.

18. An aqueous pharmaceutical preparation according to any one of claims 1 to 17, further comprising a chelating agent selected from the group consisting of ethylenediaminetetraacetic acid (EDTA), diethylenetrimenpentaacetic acid (DTPA), salts thereof, and any combination thereof.

19. The aqueous pharmaceutical formulation according to claim 18, wherein the chelating agent comprises one or both of 10 μM EDTA and 10 μM DTPA.

20. The antibody or its antigen-binding fragment in a concentration of 28 mg / mL to 138 mg / mL, 10 mM ± 1.5 mM L-histidine or histidine hydrochloride, Sucrose of 220 mM ± 33 mM, 0.06% (w / v) polysorbate 80, Water and, The aqueous pharmaceutical preparation according to claim 1, wherein the pH of the aqueous pharmaceutical preparation is approximately 5.8 to approximately 6.

2.

21. The aqueous pharmaceutical formulation according to claim 20, further comprising 10 μM EDTA or 10 μM DTPA.

22. The aqueous pharmaceutical formulation according to claim 20, further comprising 10 μM EDTA.

23. An aqueous pharmaceutical preparation according to any one of claims 1 to 22, contained within a container.

24. An aqueous pharmaceutical preparation according to any one of claims 1 to 23, which is suitable for subcutaneous delivery.

25. A pharmaceutical dosage form suitable for parenteral administration to mammals, comprising an aqueous pharmaceutical preparation according to any one of claims 1 to 24 in a suitable container.

26. The pharmaceutical unit dosage form according to claim 25, wherein the aqueous pharmaceutical preparation is suitable for intravenous, subcutaneous, or intramuscular administration.

27. The pharmaceutical unit dosage form according to claim 25 or 26, wherein the preferred container is a pre-filled syringe.

28. A sealed container containing an aqueous pharmaceutical preparation according to any one of claims 1 to 24.

29. The sealed container according to claim 28, which is a vial, syringe, microinfuser, pen delivery device, or auto-injector.

30. A sealed container according to claim 28 or 29, which is a single or multi-chamber syringe.

31. An anti-OX40 ligand (OX40L) antagonist antibody or its antigen-binding fragment in a concentration of approximately 28 mg / mL to approximately 138 mg / mL, Approximately 10 mM L-histidine or histidine hydrochloride, Approximately 220 mM sucrose, Approximately 0.06% (w / v) of polysorbate 80, Approximately 10 μM of EDTA, A pre-filled syringe containing water and an aqueous pharmaceutical preparation, A pre-filled syringe containing the aqueous pharmaceutical solution, with a pH of approximately 6.

0.

32. An anti-OX40 ligand (OX40L) antagonist antibody or its antigen-binding fragment in a concentration of approximately 28 mg / mL to approximately 138 mg / mL, Approximately 10 mM L-histidine or histidine hydrochloride, Approximately 220 mM sucrose, Approximately 0.06% (w / v) of polysorbate 80, Approximately 10 μM of EDTA, A pre-filled pen containing water and an aqueous pharmaceutical preparation, A pre-filled pen containing the aforementioned aqueous pharmaceutical solution, with a pH of approximately 6.

0.

33. An anti-OX40 ligand (OX40L) antagonist antibody or its antigen-binding fragment in a concentration of approximately 28 mg / mL to approximately 138 mg / mL, Approximately 10 mM L-histidine or histidine hydrochloride, Approximately 220 mM sucrose, Approximately 0.06% (w / v) of polysorbate 80, Approximately 10 μM of EDTA, An automatic syringe containing water and an aqueous pharmaceutical preparation, An automatic syringe in which the pH of the aqueous medicine is approximately 6.

0.

34. A pre-filled syringe according to claim 31, a pre-filled pen according to claim 32, or an auto-injector according to claim 33, wherein the antibody or its antigen-binding fragment is present at a concentration of 62.5 mg / mL or 125 mg / mL.

35. The anti-OX40 ligand (OX40L) antagonist antibody or its antigen-binding fragment (a) Heavy chain complementarity determination region (HCDR) 1 of SEQ ID NO: 42, HCDR 2 of SEQ ID NO: 44, and HCDR 3 of SEQ ID NO: 46, and Light chain complementarity determination region (LCDR) 1 of SEQ ID NO: 56, LCDR 2 of SEQ ID NO: 58, and LCDR 3 of SEQ ID NO: 60, or (b) Heavy chain complementarity determination region (HCDR) 1 of SEQ ID NO: 36, HCDR 2 of SEQ ID NO: 38, and HCDR 3 of SEQ ID NO: 40, and Light chain complementarity determination region (LCDR) 1 of SEQ ID NO: 50, LCDR 2 of SEQ ID NO: 52, and LCDR 3 of SEQ ID NO: 54, or (c) A VH domain having at least 80% identity with the amino acid sequence of SEQ ID NO: 34, and a VL domain having at least 80% identity with the amino acid sequence of SEQ ID NO: 48, or (d) A VH domain comprising a VL domain having at least 80% identity with the amino acid sequence of SEQ ID NO: 34 and / or having at least 80% identity with the amino acid sequence of SEQ ID NO: 48, or (e) A pre-filled syringe according to claim 31 or 34, or a pre-filled pen according to claim 32 or 34, or an auto-injector according to claim 33 or 34, comprising a heavy chain amino acid sequence comprising the amino acid sequence of SEQ ID NO: 62 and / or a light chain amino acid sequence comprising the amino acid sequence of SEQ ID NO:

64.

36. A pre-filled syringe according to any one of claims 31, 34, or 35, or a pre-filled pen according to any one of claims 32, 34, or 35, or an auto-injector according to any one of claims 33 to 35, comprising amliterimab or a variant thereof in an amount of approximately 31 mg / mL to approximately 125 mg / mL, preferably approximately 62.5 mg / mL or approximately 125 mg / mL.

37. A kit comprising a sealed container according to any one of claims 28 to 30, a pre-filled syringe according to any one of claims 31 or 34 to 36, or a pre-filled pen according to any one of claims 32 or 34 to 36, or an auto-injector according to any one of claims 33 to 36.

38. A sealed container comprising an aqueous pharmaceutical preparation according to any one of claims 1 to 24, A kit comprising the aqueous pharmaceutical formulation and at least one separate injection device for delivering it to a mammalian subject in need.

39. The kit according to claim 38, wherein the injection device is a syringe, a microinfuser, a pen delivery device, or an auto-injector.

40. The kit according to claim 39, wherein the injection device is a single or multi-chamber syringe.

41. An aqueous pharmaceutical formulation according to any one of claims 1 to 24, a pharmaceutical unit dosage form according to any one of claims 25 to 27, a sealed container according to any one of claims 28 to 30, a pre-filled syringe according to claim 31 or any one of claims 34 to 36, a pre-filled pen according to claim 32 or any one of claims 34 to 36, an auto-injector according to any one of claims 33 to 36, or a kit according to any one of claims 37 to 40.

42. An aqueous pharmaceutical formulation according to any one of claims 1 to 24, a pharmaceutical unit dosage form according to any one of claims 25 to 27, a sealed container according to any one of claims 28 to 30, a pre-filled syringe according to any one of claims 31 or 34 to 36, a pre-filled pen according to any one of claims 32 or 34 to 36, an auto-injector according to any one of claims 33 to 36, or a kit according to any one of claims 37 to 40, for use in the treatment of atopic dermatitis.

43. An aqueous pharmaceutical formulation according to any one of claims 1 to 24, a pharmaceutical unit dosage form according to any one of claims 25 to 27, a sealed container according to any one of claims 28 to 30, a pre-filled syringe according to any one of claims 31 or 34 to 36, a pre-filled pen according to any one of claims 32 or 34 to 36, an auto-injector according to any one of claims 33 to 36, or a kit according to any one of claims 37 to 40, for use in the treatment of asthma.

44. Use of an aqueous pharmaceutical formulation according to any one of claims 1 to 24, a pharmaceutical unit dosage form according to any one of claims 25 to 27, a sealed container according to any one of claims 28 to 30, a pre-filled syringe according to any one of claims 31 or 34 to 36, a pre-filled pen according to any one of claims 32 or 34 to 36, an auto-injector according to any one of claims 33 to 36, or a kit according to any one of claims 37 to 40, in the manufacture of a pharmaceutical for treating a disease or condition in a subject selected from the group consisting of autoimmune diseases or conditions, inflammatory diseases or conditions, systemic inflammatory diseases or conditions, and transplant rejection diseases or conditions, mediated by hOX40L.

45. Use of an aqueous pharmaceutical formulation according to any one of claims 1 to 24, a pharmaceutical unit dosage form according to any one of claims 25 to 27, a sealed container according to any one of claims 28 to 30, a pre-filled syringe according to any one of claims 31 or 34 to 36, a pre-filled pen according to any one of claims 32 or 34 to 36, an auto-injector according to any one of claims 33 to 36, or a kit according to any one of claims 37 to 40, in the manufacture of a pharmaceutical for the treatment of atopic dermatitis.

46. Use of an aqueous pharmaceutical formulation according to any one of claims 1 to 24, a pharmaceutical unit dosage form according to any one of claims 25 to 27, a sealed container according to any one of claims 28 to 30, a pre-filled syringe according to any one of claims 31 or 34 to 36, a pre-filled pen according to any one of claims 32 or 34 to 36, an auto-injector according to any one of claims 33 to 36, or a kit according to any one of claims 37 to 40, in the manufacture of a pharmaceutical for the treatment of asthma.

47. A method for treating a disease or condition in a subject selected from the group consisting of autoimmune diseases or conditions, inflammatory diseases or conditions, systemic inflammatory diseases or conditions, and transplant rejection diseases or conditions (e.g., autoimmune diseases or conditions, inflammatory diseases or conditions, systemic inflammatory diseases or conditions, and transplant rejection diseases or conditions mediated by hOX40L), comprising administering to the subject an effective amount of the aqueous pharmaceutical preparation described in any one of claims 1 to 24.

48. A method for treating atopic dermatitis, comprising administering to a subject an effective amount of an aqueous pharmaceutical preparation described in any one of claims 1 to 24.

49. A method for treating asthma, comprising administering to a subject an effective amount of an aqueous pharmaceutical preparation described in any one of claims 1 to 24.

50. (a) an anti-OX40 ligand (OX40L) antagonist antibody or antigen-binding fragment, (b) Stabilizers, (c) Chelating agent, (d) Surfactants and (e) an aqueous pharmaceutical preparation comprising a buffer containing histidine or a histidine salt, The pH of the aqueous pharmaceutical preparation is in the range of 5.5 to approximately 6.

5. The antibody or its antigen-binding fragment A VH domain comprising the HCDR1 sequence of sequence number 36 or 42, the HCDR2 sequence of sequence number 38 or 44, and the HCDR3 sequence of sequence number 40 or 46, A VL domain comprising the LCDR1 sequence of sequence number 50 or 56, the LCDR2 sequence of sequence number 52 or 58, and the LCDR3 sequence of sequence number 54 or 60, The aqueous pharmaceutical formulation is suitable for parenteral administration to mammals.

51. The aqueous pharmaceutical formulation according to claim 50, wherein the VL domain comprises the amino acid sequence of SEQ ID NO: 48, and the VH domain comprises the amino acid sequence of SEQ ID NO:

34.

52. The aqueous pharmaceutical preparation according to claim 50, wherein the antibody comprises a heavy chain and a light chain, the amino acid sequence of the heavy chain consists of the amino acid sequence of SEQ ID NO: 62, and the amino acid sequence of the light chain consists of the amino acid sequence of SEQ ID NO: 64.

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