High concentration anti-abeta protofibril antibody formulations and methods of use thereof

Abstract

Provided herein are aqueous pharmaceutical formulations comprising high concentrations of an isolated anti-Aβ fibril antibody or fragment thereof, e.g., BAN2401, arginine, polysorbate 80, and a pharmaceutically acceptable buffer, that bind to human Aβ protofibrils.

Description

[0001] This application claims priority to U.S. Provisional Application No. 62 / 992,746, filed March 20, 2020, and U.S. Provisional Application No. 63 / 027,263, filed May 19, 2020; the entire contents of both are incorporated herein by reference.

[0002] Alzheimer's disease (AD) is a progressive, neurodegenerative disorder of unknown etiology and the most common form of dementia in older adults. In 2006, there were 26.6 million cases of AD worldwide (range: 11.4-59.4 million) (Brookmeyer, R. et al., Forecasting the global burden of Alzheimer's Disease. Alzheimer Dement. 2007; 3:186-91), and it was also reported that more than 5 million people in the United States had AD (2010 Alzheimer's disease facts and figures. Alzheimer Dement. 2010; 6:158-94). By 2050, the global prevalence of Alzheimer's disease (AD) is projected to reach 106.8 million (range: 47.2 million–221.2 million), with an estimated prevalence of 11 million to 16 million in the United States alone. (Brookmeyer, see above, and 2010 Alzheimer's disease facts and figures, see above).

[0003] The disease typically involves a general decline in cognitive function, which progresses slowly and leaves terminally ill patients bedridden. Patients with Alzheimer's disease (AD) usually survive only 3 to 10 years after symptom onset, although known extremes are 2 to 20 years. (Hebert, LE, et al., Alzheimer's disease in the US population: prevalence estimates using the 2000 census. ArchNeurol. 2003; 60:1119-1122.) Although deaths due to AD are significantly underestimated because death certificates rarely attribute death to AD, AD remains the seventh leading cause of death in the United States and the fifth leading cause of death among Americans over 65 years of age. (2010 Alzheimer's disease facts and figures, see above.)

[0004] Histologically, the disease is characterized by neuroinflammatory plaques, primarily found in the associative cortex, limbic system, and basal ganglia. The main component of these plaques is amyloid-β peptide (Aβ). Aβ exists in various conformational states: monomers, oligomers, primary fibrils, and insoluble filaments. The mechanistic relationship between Alzheimer's disease pathogenesis and Aβ production remains unclear. However, several anti-Aβ antibodies are currently undergoing clinical trials as potential therapeutic agents for Alzheimer's disease.

[0005] Anti-Aβ antibodies and other proteins can be administered to subjects via intravenous, subcutaneous, intramuscular, and other routes. The dosage and / or formulation of the antibody can present numerous challenges in developing a suitable drug formulation. For example, at high antibody concentrations, antibody stability can be problematic due to the formation or fragmentation of protein-protein aggregates. Typically, aggregation increases with increasing antibody concentration. Furthermore, high-concentration antibody formulations require high concentrations of stabilizers and other excipients to achieve long-term protein stability and shelf life. High-concentration antibody formulations are also often viscous, which can complicate the manufacture and administration of the drug formulation.

[0006] This article provides pharmaceutical formulations comprising a therapeutically effective amount of at least one isolated anti-Aβ primary fibrillation antibody or fragment thereof that binds to Aβ primary fibrillation. This article also provides pharmaceutical formulations comprising 80-300 mg / mL of an isolated anti-Aβ primary fibrillation antibody or fragment thereof that binds to Aβ primary fibrillation, wherein the antibody is BAN2401 (also known as lecanemab). The pharmaceutical formulations provided herein have been found to be advantageous. For example, despite high concentrations of the anti-Aβ primary fibrillation antibody (e.g., 100 mg / mL or 200 mg / mL), the protein-protein aggregation rate is unexpectedly low and similar to that typically observed at much lower antibody concentrations (e.g., 10 mg / mL). In some embodiments, the currently disclosed pharmaceutical formulation exhibits a lower initial aggregate rate (e.g., an initial aggregate level of about 0.3% for 100 mg / mL of anti-Aβ profibrillary antibody compared to formulations having significantly lower concentrations of anti-Aβ profibrillary antibody, compared to about 0.8% for 10 mg / mL of anti-Aβ profibrillary antibody). In some embodiments, the currently disclosed pharmaceutical formulation exhibits a lower rate of subvisible particle formation (e.g., 10.6 particles / mL for 100 mg / mL of anti-Aβ profibrillary antibody compared to 12.6 particles / mL for 10 mg / mL of anti-Aβ profibrillary antibody) compared to formulations having significantly lower concentrations of anti-Aβ profibrillary antibody. In some embodiments, the currently disclosed pharmaceutical formulation exhibits reduced aggregation rate, reduced initial aggregation level, reduced protein fragmentation rate, and / or reduced subvisible particle formation compared to formulations having significantly lower concentrations of anti-Aβ profibrillary antibody. Low aggregation rates, initial aggregation levels, protein fragmentation rates, and / or the generation of subvisible particles may allow for increased stability and / or extended product shelf life. Furthermore, the excipients in the currently disclosed pharmaceutical formulations may be present in lower amounts compared to currently marketed intravenous products. In some embodiments, the pH and osmolality of the currently disclosed pharmaceutical formulations are acceptable for intravenous administration after dilution in an intravenous solution.

[0007] In some embodiments, the at least one anti-Aβ protofibrillary antibody comprises (i) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:1, and (ii) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:2.

[0008] In some embodiments, the at least one anti-Aβ protofibrillary antibody comprises three heavy chain complementarity-determining regions (HCDR1, HCDR2, and HCDR3) containing the amino acid sequences of SEQ ID NO:5 (HCDR1), SEQ ID NO:6 (HCDR2), and SEQ ID NO:7 (HCDR3); and three light chain complementarity-determining regions (LCDR1, LCDR2, and LCDR3) containing the amino acid sequences of SEQ ID NO:8 (LCDR1), SEQ ID NO:9 (LCDR2), and SEQ ID NO:10 (LCDR3).

[0009] The amino acid assignment of each domain is usually based on the definition of the sequence of the protein of immunological interest (SEQUENCES OF PROTEINS OF IMMUNOLOGICAL INTEREST) ​​(Kabat et al., 5th edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242, 1991, hereinafter referred to as the "Kabat Report").

[0010] In some embodiments, the at least one anti-Aβ primary fibrillary antibody comprises a human constant region. In some embodiments, the human constant region of the at least one anti-Aβ primary fibrillary antibody comprises a heavy chain constant region selected from IgG1, IgG2, IgG3, IgG4, IgM, IgA, IgE, and any allelic variants thereof as disclosed in the Kabat report. Any one or more of such sequences may be used in this disclosure. In some embodiments, the heavy chain constant region is selected from IgG1 and its allelic variants. The amino acid sequence of the human IgG1 constant region is known in the art and is shown in SEQ ID NO:3.

[0011] In some embodiments, the human constant region of the at least one anti-Aβ antibody comprises a light chain constant region selected from the κ-λ chain constant region and any allelic variant thereof as discussed in the Kabat report. Any one or more of such sequences may be used in this disclosure. In some embodiments, the light chain constant region is selected from κ and its allelic variants. The amino acid sequence of the human κ chain constant region is known in the art and is shown in SEQ ID NO:4.

[0012] In some embodiments, the at least one anti-Aβ primary fibrillary antibody comprises a human heavy chain and light chain variable region framework. In some embodiments, the at least one anti-Aβ primary fibrillary antibody comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprising the amino acid sequence of SEQ ID NO:1 and the light chain variable region comprising the amino acid sequence of SEQ ID NO:2. In some embodiments, the at least one anti-Aβ primary fibrillary antibody comprises a human IgG1 heavy chain constant region and a human Igκ light chain constant region. In some embodiments, the at least one anti-Aβ primary fibrillary antibody comprises a heavy chain constant region and a light chain constant region, the heavy chain constant region comprising the amino acid sequence of SEQ ID NO:3 and the light chain constant region comprising the amino acid sequence of SEQ ID NO:4.

[0013] In some embodiments, the at least one anti-Aβ primary fibrillary antibody is BAN2401, also known as lecanemab. BAN2401 is a humanized IgG1 monoclonal form of mAb158, a murine monoclonal antibody produced to target primary fibrillary fibers and disclosed in WO2007 / 108756 and the Journal of Alzheimer's Disease 43:575-588 (2015). BAN2401 is at least one anti-Aβ primary fibrillary antibody that exhibits low affinity for Aβ monomers while binding to soluble Aβ aggregates with high selectivity. For example, BAN2401 has been reported to exhibit selectivity for soluble Aβ primary fibrillary fibers that is approximately 1000-fold higher than that for Aβ monomers and 5 to 10-fold higher than that for insoluble Aβ fibrils.

[0014] BAN2401 comprises (i) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:1, and (ii) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:2. The full-length sequences of the heavy and light chains of BAN2401 are shown in SEQ ID NO:11 and 12, and described in WO 2007 / 108756 and Journal of Alzheimer's Disease 43:575-588 (2015).

[0015] Other non-limiting examples of antibodies applicable as at least one anti-Aβ protofibrillary antibody in this disclosure include the antibodies disclosed in WO 2002 / 003911, WO 2005 / 123775, WO2007 / 108756, WO 2011 / 001366, WO 2011 / 104696 and WO2016 / 005466.

[0016] In some embodiments, the isolated anti-Aβ primary fibrillary antibody is present at a concentration of at least 80 mg / mL. In some embodiments, the isolated anti-Aβ primary fibrillary antibody is present at a concentration of at least 100 mg / mL. In some embodiments, the isolated anti-Aβ primary fibrillary antibody is present at a concentration of at least 200 mg / mL. In some embodiments, the isolated anti-Aβ primary fibrillary antibody is present at a concentration of at least 250 mg / mL. In some embodiments, the isolated antibody or fragment thereof is present at a concentration ranging from 80 mg / mL to 300 mg / mL. In some embodiments, the isolated anti-Aβ primary fibrillary antibody is present at a concentration ranging from 85 mg / mL to 275 mg / mL. In some embodiments, the isolated anti-Aβ primary fibrillary antibody is present at a concentration ranging from 90 mg / mL to 250 mg / mL. In some embodiments, the isolated anti-Aβ primary fibrillary antibody is present at a concentration ranging from 95 mg / mL to 225 mg / mL. In some embodiments, the isolated anti-Aβ protofibrillary antibody is present at a concentration ranging from 100 mg / mL to 200 mg / mL. In some embodiments, the isolated antibody or its fragment is present at concentrations of 80 mg / mL, 90 mg / mL, 100 mg / mL, 110 mg / mL, 120 mg / mL, 130 mg / mL, 140 mg / mL, 150 mg / mL, 160 mg / mL, 170 mg / mL, 180 mg / mL, 190 mg / mL, 200 mg / mL, 210 mg / mL, 220 mg / mL, 230 mg / mL, 240 mg / mL, 250 mg / mL, 260 mg / mL, 270 mg / mL, 280 mg / mL, 290 mg / mL, or 300 mg / mL. In some embodiments, the isolated antibody or its fragment is present at a concentration of 100 mg / mL. In some embodiments, the isolated antibody or its fragment is present at a concentration of 200 mg / mL. In some embodiments, the isolated antibody or fragment thereof is present at a concentration of 250 mg / mL. In some embodiments, the isolated antibody or fragment thereof is present at a concentration of 300 mg / mL. In some embodiments, the isolated antibody or fragment thereof is BAN2401. In some embodiments, the isolated antibody or fragment thereof comprises (i) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:1, and (ii) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:2.In some embodiments, the isolated antibody or fragment thereof comprises three heavy chain complementarity-determining regions (HCDR1, HCDR2, and HCDR3) containing the amino acid sequences of SEQ ID NO:5 (HCDR1), SEQ ID NO:6 (HCDR2), and SEQ ID NO:7 (HCDR3); and three light chain complementarity-determining regions (LCDR1, LCDR2, and LCDR3) containing the amino acid sequences of SEQ ID NO:8 (LCDR1), SEQ ID NO:9 (LCDR2), and SEQ ID NO:10 (LCDR3).

[0017] In some embodiments, a pharmaceutical formulation comprising a therapeutically effective amount of at least one isolated anti-Aβ primary fibrillary antibody or fragment thereof that binds to Aβ primary fibrils further comprises at least one additional component. In some embodiments, the at least one additional component in the pharmaceutical formulation is selected from buffers. In some embodiments, the buffer is a citrate buffer. In some embodiments, the buffer is a histidine buffer. In some embodiments, the at least one additional component in the pharmaceutical formulation is selected from emulsifiers. In some embodiments, the at least one additional component in the pharmaceutical formulation is selected from citric acid (or citrate monohydrate), sodium chloride, histidine (and / or histidine hydrochloride), arginine (and / or arginine hydrochloride), and polysorbate 80. In some embodiments, the at least one additional component in the pharmaceutical formulation is selected from citric acid (and / or citrate monohydrate), arginine (and / or arginine hydrochloride), and polysorbate 80. In some embodiments, the at least one additional component in the pharmaceutical formulation is selected from histidine (and / or histidine hydrochloride), arginine (and / or arginine hydrochloride), and polysorbate 80.

[0018] In some embodiments, the pharmaceutical formulation comprises arginine (and / or arginine hydrochloride). In some embodiments, the concentration of arginine (and / or arginine hydrochloride) in the pharmaceutical formulation ranges from about 100 mM to about 400 mM. In some embodiments, the concentration of arginine (and / or arginine hydrochloride) in the pharmaceutical formulation ranges from about 110 mM to about 380 mM. In some embodiments, the concentration of arginine (and / or arginine hydrochloride) in the pharmaceutical formulation ranges from about 120 mM to about 360 mM. In some embodiments, the concentration of arginine (and / or arginine hydrochloride) in the pharmaceutical formulation ranges from about 125 mM to about 350 mM. In some embodiments, the concentration of arginine (and / or arginine hydrochloride) in the pharmaceutical formulation is 125 mM. In some embodiments, the concentration of arginine (and / or arginine hydrochloride) in the pharmaceutical formulation is 200 mM. In some embodiments, the concentration of arginine (and / or arginine hydrochloride) in the pharmaceutical formulation is 350 mM.

[0019] In some embodiments, the pharmaceutical preparation comprises histidine. In some embodiments, the concentration of histidine in the pharmaceutical preparation ranges from about 10 mM to about 100 mM. In some embodiments, the concentration of histidine in the pharmaceutical preparation ranges from 10 mM to 100 mM, 12 mM to 80 mM, 14 mM to 60 mM, 15 mM to 55 mM, 15 mM to 35 mM, or 15 mM to 25 mM. In some embodiments, the concentration of histidine is 25 mM. In some embodiments, the concentration of histidine is 50 mM.

[0020] In some embodiments, the pharmaceutical formulation comprises polysorbate 80. In some embodiments, the concentration of polysorbate 80 in the pharmaceutical formulation ranges from about 0.01% w / v to 0.1% w / v, 0.01% w / v to 0.08% w / v, 0.02% w / v to 0.08% w / v, 0.03% w / v to 0.07% w / v, or 0.04% w / v to 0.06% w / v. In some embodiments, the polysorbate 80 is present in the pharmaceutical formulation at concentrations of 0.01% w / v, 0.02% w / v, 0.03% w / v, 0.04% w / v, 0.05% w / v, 0.06% w / v, 0.07% w / v, or 0.08% w / v. In some embodiments, the polysorbate 80 is present in the pharmaceutical formulation at a concentration of 0.02% w / v. In some embodiments, the polysorbate 80 is present in the pharmaceutical preparation at a concentration of 0.05% w / v.

[0021] In some embodiments, the pharmaceutical preparation comprises citrate monohydrate. In some embodiments, the concentration of citrate monohydrate in the pharmaceutical preparation ranges from about 10 mM to 100 mM. In some embodiments, the concentration of citrate monohydrate in the pharmaceutical preparation ranges from 10 mM to 100 mM, 10 mM to 90 mM, 15 mM to 85 mM, 20 mM to 80 mM, 25 mM to 75 mM, 30 mM to 70 mM, 30 mM to 60 mM, or 30 mM to 50 mM. In some embodiments, the concentration of citrate monohydrate in the pharmaceutical preparation is 50 mM.

[0022] In some embodiments, this disclosure provides a pharmaceutical preparation having a pH range of 4.5 to 5.5. In some embodiments, the pH range of the pharmaceutical preparation is 4.0 to 6.0, 4.2 to 5.8, 4.3 to 5.7, 4.4 to 5.6, or 4.5 to 5.5. In some embodiments, the pH is 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, or 5.5. In some embodiments, the pH is 5.0.

[0023] In some embodiments, the pharmaceutical formulations disclosed herein may be in the form of solutions and / or any other suitable liquid formulations that a person skilled in the art would consider appropriate. The routes of administration for the compositions disclosed herein may be intravenous or subcutaneous. In some embodiments, the pharmaceutical formulation is formulated as a sterile, pyrogen-free liquid for intravenous administration. In some embodiments, the pharmaceutical formulation is formulated as a sterile, pyrogen-free liquid for subcutaneous administration. In some embodiments, the pharmaceutical composition is a saline solution.

[0024] In some embodiments, the pharmaceutical formulation is a liquid dosage form comprising at least one isolated anti-Aβ primary fibrillary antibody or fragment thereof (e.g., BAN2401) that binds to Aβ primary fibrillary cells, and further comprising, for example, citrate monohydrate, arginine, arginine hydrochloride, and polysorbate 80. In some embodiments, the pharmaceutical formulation comprises 100 mg / mL of at least one isolated anti-Aβ primary fibrillary antibody or fragment thereof (e.g., BAN2401) that binds to Aβ primary fibrillary cells, 50 mM of citrate monohydrate, 110 mM of arginine, 240 mM of arginine hydrochloride, and 0.05% (w / v) of polysorbate 80, and a pH of 5.0 ± 0.4.

[0025] In some embodiments, the pharmaceutical formulation is a liquid dosage form comprising at least one isolated anti-Aβ primary fibrillary antibody or fragment thereof (e.g., BAN2401) that binds to Aβ primary fibrillary cells, and further comprising, for example, histidine, histidine hydrochloride, arginine hydrochloride, and polysorbate 80. In some embodiments, the pharmaceutical formulation comprises 100 mg / mL or 200 mg / mL of at least one isolated anti-Aβ primary fibrillary antibody or fragment thereof (e.g., BAN2401) that binds to Aβ primary fibrillary cells, 25 mM of histidine and histidine hydrochloride, 200 mM of arginine hydrochloride, and 0.05% (w / v) of polysorbate 80, and a pH of 5.0 ± 0.4.

[0026] In some embodiments, the pharmaceutical formulation is a liquid dosage form comprising at least one isolated anti-Aβ primary fibrillary antibody or fragment thereof (e.g., BAN2401) that binds to Aβ primary fibrillary cells, and further comprising, for example, histidine, histidine hydrochloride, arginine hydrochloride, and polysorbate 80. In some embodiments, the pharmaceutical formulation comprises 200 mg / mL of at least one isolated anti-Aβ primary fibrillary antibody or fragment thereof (e.g., BAN2401) that binds to Aβ primary fibrillary cells, 50 mM of histidine and histidine hydrochloride, 125 mM of arginine hydrochloride, and 0.02% (w / v) of polysorbate 80, and a pH of 5.0 ± 0.4.

[0027] In some embodiments, the pharmaceutical formulation is a liquid dosage form comprising at least one isolated anti-Aβ primary fibrillary antibody or fragment thereof (e.g., BAN2401) that binds to Aβ primary fibrillary cells, and further comprising, for example, histidine, histidine hydrochloride, arginine hydrochloride, and polysorbate 80. In some embodiments, the pharmaceutical formulation comprises 200 mg / mL of at least one isolated anti-Aβ primary fibrillary antibody or fragment thereof (e.g., BAN2401) that binds to Aβ primary fibrillary cells, 50 mM of citric acid (and / or citric acid monohydrate), 125 mM of arginine (and / or arginine hydrochloride), and 0.02% (w / v) of polysorbate 80, and a pH of 5.0 ± 0.4.

[0028] BAN2401 and the methods including the use of BAN2401 are disclosed in U.S. Provisional Application No. 62 / 749,614 and PCT International Application No. PCT / US 2019 / 043067, the entire contents of which are incorporated herein by reference. Attached Figure Description

[0029] Figure 1 The percentage of aggregation in formulations of BAN2401 at concentrations of 10 mg / mL, 100 mg / mL, and 200 mg / mL was described at 5°C at time points of 0, 1, 2, and 3 months.

[0030] Figure 2 The percentage of fragmentation in formulations of BAN2401 at concentrations of 10 mg / mL, 100 mg / mL, and 200 mg / mL was described at 5°C at time points of 0, 1, 2, and 3 months.

[0031] Figure 3 The percentage of monomers in formulations of BAN2401 at concentrations of 10 mg / mL, 100 mg / mL, and 200 mg / mL was described at 0, 1, 2, and 3 months at 5°C.

[0032] Figure 4 The percentage of aggregation in formulations of BAN2401 at concentrations of 10 mg / mL, 100 mg / mL, and 200 mg / mL was described at 25°C at time points of 0, 1, 2, and 3 months.

[0033] Figure 5 The percentage of fragmentation in formulations of BAN2401 at concentrations of 10 mg / mL, 100 mg / mL, and 200 mg / mL was described at 25°C at time points of 0, 1, 2, and 3 months.

[0034] Figure 6The percentage of monomers in formulations of BAN2401 at concentrations of 10 mg / mL, 100 mg / mL, and 200 mg / mL was described at 0, 1, 2, and 3 months at 25°C.

[0035] Figure 7 The percentage of aggregation in formulations of BAN2401 at 5°C and at pH values ​​of 4.5, 5.0, 5.5, 6.0, and 6.5 at 0, 1, 2, and 3 months is described.

[0036] Figure 8 The percentage of fragmentation in BAN2401 formulations at pH 4.5, 5.0, 5.5, 6.0, and 6.5 at 5°C for 0, 1, 2, and 3 months is described.

[0037] Figure 9 The percentage of monomers in BAN2401 formulations at pH 4.5, 5.0, 5.5, 6.0, and 6.5 at 0, 1, 2, and 3 months at 5°C is described.

[0038] Figure 10 The percentage of aggregation in formulations of BAN2401 at 25°C and at pH values ​​of 4.5, 5.0, 5.5, 6.0, and 6.5 at 0, 1, 2, and 3 months is described.

[0039] Figure 11 The percentage of fragmentation in BAN2401 formulations at 25°C and at pH values ​​of 4.5, 5.0, 5.5, 6.0, and 6.5 at 0, 1, 2, and 3 months is described.

[0040] Figure 12 The percentage of monomers in BAN2401 formulations at pH 4.5, 5.0, 5.5, 6.0, and 6.5 at 25°C for 0, 1, 2, and 3 months is described.

[0041] Figure 13 The percentage of aggregation in formulations of BAN2401 at concentrations of 10 mg / mL, 100 mg / mL, and 200 mg / mL was described at 0, 1, and 2 months at 25°C.

[0042] Figure 14 The percentage of fragmentation in formulations of BAN2401 at concentrations of 10 mg / mL, 100 mg / mL, and 200 mg / mL was described at 0, 1, and 2 months at 25°C.

[0043] Figure 15The percentage of monomers in formulations of BAN2401 at concentrations of 10 mg / mL, 100 mg / mL, and 200 mg / mL was described at 0, 1, and 2 months at 25°C.

[0044] Figure 16 The percentage of monomer aggregation in BAN2401 formulations was described as a function of arginine concentration.

[0045] Figure 17 A flowchart describing the process of manufacturing a 10 mg / mL injection of BAN2401 is provided.

[0046] Figure 18 A flowchart describing the process of manufacturing a 100 mg / mL injection of BAN2401 is provided.

[0047] Figure 19 The pH values ​​of the formulation of BAN2401 at 5°C and at 0 and 12 months are described.

[0048] Figure 20 The pH values ​​of BAN2401 formulations at 25°C for 0, 1, and 3 months are described.

[0049] Figure 21 The pH values ​​of BAN2401 formulations after one month at 5°C and with stirring at 25°C are described.

[0050] Figure 22 The absorbance of the formulation of BAN2401 at 405 nm was described at 5 °C for 0, 1, 3, 6, 9, and 12 months.

[0051] Figure 23 The absorbance of the formulation of BAN2401 at 405 nm was described at 25 °C for 0, 1, and 3 months.

[0052] Figure 24 The absorbance of the BAN2401 formulation at 405 nm was described after one month at 5 °C and under stirring at 25 °C.

[0053] Figure 25 The percentage of aggregation in the formulation of BAN2401 is described at 5°C at time points of 0, 1, 3, 6, 9, and 12 months.

[0054] Figure 26 The percentage of fragmentation in the formulation of BAN2401 is described at 5°C at time points of 0, 1, 3, 6, 9, and 12 months.

[0055] Figure 27The percentage of monomers in the formulation of BAN2401 is described at 5°C for 0, 1, 3, 6, 9, and 12 months.

[0056] Figure 28 The percentage of aggregation in the formulation of BAN2401 is described at 0, 1, and 3 months at 25°C.

[0057] Figure 29 The percentage of fragmentation in the formulation of BAN2401 is described at 0, 1, and 3 months at 25°C.

[0058] Figure 30 The percentage of monomers in the formulation of BAN2401 is described at 0, 1, and 3 months at 25°C.

[0059] Figure 31 The percentage of aggregation in the formulation of BAN2401 is described at 5°C for one month and at 25°C with stirring.

[0060] Figure 32 The percentage of fragmentation in BAN2401 formulations was described at 5°C for one month and at 25°C with stirring.

[0061] Figure 33 The percentage of monomers in BAN2401 formulations is described at 5°C for one month and at 25°C with stirring. definition

[0062] The following are definitions of the terms used in this application.

[0063] Unless the context clearly indicates otherwise, as used herein, the singular terms “a / an” and “the” include plural references.

[0064] As used herein, the phrase “and / or” means “any one or both” of the elements so combined, that is, elements that are combined in some cases and not combined in others. Therefore, as a non-limiting example, when “A and / or B” is used in conjunction with open-ended terms such as “including,” in some embodiments it may refer only to A (optionally including elements other than B); in other embodiments it may refer only to B (optionally including elements other than A); in still other embodiments it may refer to both A and B (optionally including other elements); and so on.

[0065] As used herein, “at least one” means one or more elements in the list of elements, but is not required to include at least one of each element specifically listed in the list of elements, and does not exclude any combination of elements in the list of elements. This definition also allows for the optional presence of elements other than those specifically identified in the list of elements referred to by the phrase “at least one,” whether related to or unrelated to those specifically identified elements. Thus, as a non-limiting example, “at least one of A and B” (or equivalently, “at least one of A or B”, or equivalently, “at least one of A and / or B”) may, in one embodiment, mean at least one (optionally including more than one) A without B (and optionally including elements other than B); in another embodiment, mean at least one (optionally including more than one) B without A (and optionally including elements other than A); in yet another embodiment, mean at least one (optionally including more than one) A and at least one (optionally including more than one) B (and optionally including other elements); and so on.

[0066] When a number is described alone or as part of a numerical range, it should be understood that the value may vary reasonably above or below the value, as would be recognized by those skilled in the art.

[0067] As used herein, an antibody “fragment” comprises a portion of the antibody, such as containing an antigen-binding region or its variable region. Non-limiting examples of fragments include Fab fragments, Fab' fragments, F(ab')2 fragments, Fv fragments, biantibodies, linear antibodies, and single-chain antibody molecules.

[0068] As used herein, “fragmentation” or “fragmentation” refers to the degradation of an antibody or a fragment thereof when it is present in or added to a preparation. Fragments resulting from fragmentation or fragmentation may or may not bind to antigens to which the antibody or its fragments are bound.

[0069] As used herein, "histidine buffer" may comprise histidine, histidine hydrochloride, or a combination thereof, wherein histidine hydrochloride may be histidine hydrochloride monohydrate.

[0070] As used herein, a “citrate buffer” may contain citric acid, its salts, or combinations thereof, wherein the citric acid may be citric acid monohydrate or anhydrous citric acid. Non-limiting embodiments disclosed herein:

[0071] Some embodiments disclosed herein relate to aqueous pharmaceutical formulations.

[0072] In some embodiments, an aqueous pharmaceutical preparation is provided, the aqueous pharmaceutical preparation comprising: (a) An isolated anti-Aβ primary fibrillary antibody or fragment thereof, in concentrations of 80-300 mg / ml, comprising (i) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:1, and (ii) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:2; and (b) 100 mM to 400 mM of arginine, wherein the arginine is arginine, arginine hydrochloride, or a combination thereof; the aqueous pharmaceutical preparation optionally comprises any one or more of the following: a. 0.01% w / v to 0.1% w / v of polysorbate 80; b. A pharmaceutically acceptable buffer, optionally wherein the buffer is a histidine buffer or a citrate buffer; and c. pH 4.5 to 5.5.

[0073] In some embodiments, an aqueous pharmaceutical preparation is provided, the aqueous pharmaceutical preparation comprising: (a) An isolated anti-Aβ primary fibrillary antibody or fragment thereof, in concentrations of 80-300 mg / ml, comprising (i) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:1, and (ii) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:2; and (b) A pharmaceutically acceptable buffer, wherein the buffer is a histidine buffer, optionally wherein the concentration of the histidine buffer is about 25 mM; the aqueous pharmaceutical preparation optionally comprises any one or more of the following: a. 0.01% w / v to 0.1% w / v of polysorbate 80; b. 100 mM to 400 mM of arginine, arginine hydrochloride, or a combination thereof; and c. pH 4.5 to 5.5.

[0074] Some embodiments relate to aqueous pharmaceutical formulations comprising: (a) Isolated anti-Aβ primary fibrillary antibody or fragment thereof that binds to human Aβ primary fibrillary cells at a concentration of 80 mg / mL to 300 mg / mL. (b) 100mM to 400mM arginine, (c) Polysorbate 80, from 0.01% w / v to 0.1% w / v, and (d) Pharmaceutically acceptable buffers. The drug formulation has a pH range of 4.5 to 5.5, and The antibody or its fragment comprises: (i) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:1, and (ii) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:2. And the arginine in it is arginine, arginine hydrochloride, or a combination thereof.

[0075] antibody concentration

[0076] For any aqueous pharmaceutical formulation described herein, the antibody may be present at the following concentrations.

[0077] In some embodiments of the pharmaceutical formulation, the isolated anti-Aβ primary fibrillary antibody or a fragment thereof is present at a concentration of 100 mg / mL or higher.

[0078] In some embodiments of the pharmaceutical formulation, the isolated anti-Aβ primary fibrillary antibody or a fragment thereof is present at a concentration of 200 mg / mL or higher.

[0079] In some embodiments of the pharmaceutical formulation, the isolated anti-Aβ primary fibrillary antibody or a fragment thereof is present at a concentration range of 80 mg / mL to 300 mg / mL. In some embodiments of the pharmaceutical formulation, the isolated anti-Aβ primary fibrillary antibody or a fragment thereof is present at a concentration range of 80 mg / mL to 240 mg / mL.

[0080] In some embodiments of the pharmaceutical formulation, the isolated anti-Aβ primary fibrillary antibody or a fragment thereof is present at a concentration range of 100 mg / mL to 200 mg / mL.

[0081] In some embodiments of the pharmaceutical formulation, the isolated anti-Aβ primary fibrillary antibody or fragment thereof is present at concentrations ranging from 80 mg / mL to 300 mg / mL, 85 mg / mL to 275 mg / mL, 90 mg / mL to 250 mg / mL, 95 mg / mL to 225 mg / mL, and 100 to 200 mg / mL. In some embodiments of the pharmaceutical formulation, the isolated anti-Aβ primary fibrillary antibody or fragment thereof is present at concentrations ranging from 90 mg / mL to 220 mg / mL, 100 mg / mL to 210 mg / mL, or 110 mg / mL to 200 mg / mL.

[0082] In some embodiments of the pharmaceutical formulation, the isolated anti-Aβ primary fibrillary antibody or a fragment thereof is present at concentrations of 80 mg / mL, 90 mg / mL, 100 mg / mL, 110 mg / mL, 120 mg / mL, 180 mg / mL, 190 mg / mL, 200 mg / mL, 210 mg / mL, 220 mg / mL, 230 mg / mL, 240 mg / mL, 250 mg / mL, 260 mg / mL, 270 mg / mL, 280 mg / mL, 290 mg / mL, or 300 mg / mL.

[0083] In some embodiments of the pharmaceutical formulation, the isolated anti-Aβ primary fibrillary antibody or a fragment thereof is present at a concentration of 100 mg / mL.

[0084] In some embodiments of the pharmaceutical formulation, the isolated anti-Aβ primary fibrillary antibody or a fragment thereof is present at a concentration of 200 mg / mL.

[0085] In some embodiments of the pharmaceutical formulation, the isolated anti-Aβ primary fibrillary antibody or a fragment thereof is BAN2401.

[0086] Arginine

[0087] For any aqueous pharmaceutical formulation described herein, the formulation may contain arginine as follows.

[0088] In some embodiments, the pharmaceutical formulation comprises arginine. In some embodiments, the arginine is arginine, arginine hydrochloride, or a combination thereof.

[0089] In some embodiments of the pharmaceutical formulation, the concentration of arginine ranges from about 100 mM to about 400 mM.

[0090] In some embodiments of the pharmaceutical formulation, the concentration range of arginine is 100 mM to 400 mM, 110 mM to 380 mM, 120 mM to 360 mM, 125 mM to 350 mM, 100 mM to 200 mM, 125 mM to 200 mM, or 150 mM to 200 mM. In some embodiments of the pharmaceutical formulation, the concentration range of arginine is 150 mM to 250 mM, 160 mM to 240 mM, 170 mM to 230 mM, 180 mM to 220 mM, 190 mM to 210 mM, arginine hydrochloride, or a combination thereof.

[0091] In some embodiments of this pharmaceutical formulation, the concentration of arginine is 125 mM.

[0092] In some embodiments of this pharmaceutical formulation, the concentration of arginine is 200 mM.

[0093] In some embodiments of the pharmaceutical formulation, the concentration range of arginine is 200mM to 400mM, 210mM ​​to 390mM, 220mM to 380mM, 230mM to 370mM, 240mM to 360mM, 240mM to 350mM, or 250mM to 350mM.

[0094] In some embodiments of this pharmaceutical formulation, the concentration of arginine is 350 mM.

[0095] Polysorbate 80 (PS80)

[0096] For any aqueous pharmaceutical formulation described herein, the formulation may contain polysorbate 80 as follows.

[0097] In some embodiments of the pharmaceutical formulation, the concentration range of the polysorbate 80 is about 0.01% w / v to 0.1% w / v, 0.01% w / v to 0.08% w / v, 0.02% w / v to 0.08% w / v, 0.03% w / v to 0.07% w / v, or 0.04% w / v to 0.06% w / v.

[0098] In some embodiments of the pharmaceutical formulation, the polysorbate 80 is present at concentrations of 0.01% w / v, 0.02% w / v, 0.03% w / v, 0.04% w / v, 0.05% w / v, 0.06% w / v, 0.07% w / v, or 0.08% w / v.

[0099] In some embodiments of the pharmaceutical formulation, the polysorbate 80 is present at a concentration of 0.02% w / v.

[0100] In some embodiments of the pharmaceutical formulation, the polysorbate 80 is present at a concentration of 0.05% w / v.

[0101] buffer

[0102] For any aqueous pharmaceutical formulation described herein, the formulation may contain pharmaceutically acceptable buffers as follows.

[0103] In some embodiments of this pharmaceutical formulation, the pharmaceutically acceptable buffer is a citrate buffer.

[0104] In some embodiments of the pharmaceutical formulation, the citrate buffer is present at a concentration of about 10 mM to about 100 mM.

[0105] In some embodiments of the pharmaceutical formulation, the concentration range of the citrate buffer is 10 mM to 100 mM, 10 mM to 90 mM, 15 mM to 85 mM, 20 mM to 80 mM, 25 mM to 75 mM, 30 mM to 70 mM, 30 mM to 60 mM, or 30 mM to 50 mM.

[0106] In some embodiments of this pharmaceutical formulation, the citrate buffer is present at a concentration of 50 mM.

[0107] In some embodiments of this pharmaceutical formulation, the pharmaceutically acceptable buffer is a histidine buffer.

[0108] In some embodiments of the pharmaceutical formulation, the histidine buffer is present at a concentration of about 10 mM to about 100 mM.

[0109] In some embodiments of the pharmaceutical formulation, the concentration range of the histidine buffer is 10 mM to 100 mM, 12 mM to 80 mM, 14 mM to 60 mM, or 15 mM to 55 mM, 15 mM to 35 mM, or 15 mM to 25 mM.

[0110] In some embodiments of this pharmaceutical formulation, the histidine buffer is present at a concentration of 25 mM.

[0111] In some embodiments of this pharmaceutical formulation, the histidine buffer is present at a concentration of 50 mM.

[0112] In some embodiments of the pharmaceutical formulation, the histidine buffer comprises histidine and histidine hydrochloride monohydrate. In some embodiments of the pharmaceutical formulation, the histidine buffer comprises histidine and histidine hydrochloride monohydrate, wherein the concentration of histidine is about 0.1 to 0.3 mg / mL, and the concentration of histidine hydrochloride monohydrate is about 4 to 6 mg / mL. In some embodiments of the pharmaceutical formulation, the histidine buffer comprises histidine and histidine hydrochloride monohydrate, wherein the concentration of histidine is about 0.26 mg / mL, and the concentration of histidine hydrochloride monohydrate is about 4.89 mg / mL. In some embodiments of the pharmaceutical formulation, the histidine buffer comprises histidine and histidine hydrochloride monohydrate, wherein the histidine comprises 0.2-0.3 mg / mL of histidine and 4.4 to 4.9 mg / mL of histidine hydrochloride, optionally wherein the histidine hydrochloride is a monohydrate.

[0113] pH

[0114] For any aqueous pharmaceutical formulation described herein, the formulation may have the following pH values.

[0115] In some embodiments of the pharmaceutical formulation, the pH ranges from 4.0 to 6.0, 4.2 to 5.8, 4.3 to 5.7, 4.4 to 5.6, or 4.5 to 5.5.

[0116] In some embodiments of this pharmaceutical formulation, the pH is 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, or 5.5.

[0117] In some embodiments of this pharmaceutical formulation, the pH is 4.5 to 5.5.

[0118] In some embodiments of this pharmaceutical formulation, the pH is 5.0.

[0119] In some embodiments of this pharmaceutical preparation, the pharmaceutical preparation is suitable for intravenous injection.

[0120] In some embodiments of this pharmaceutical formulation, the pharmaceutical formulation is suitable for subcutaneous injection.

[0121] In some embodiments of the pharmaceutical formulation, the pharmaceutical formulation contains methionine.

[0122] In some embodiments, an aqueous pharmaceutical preparation is disclosed, the aqueous pharmaceutical preparation comprising: (a) BAN2401 at concentrations ranging from 80 mg / mL to 240 mg / mL, (b) Arginine hydrochloride at 140 mM to 260 mM (c) Polysorbate 80, from 0.01% w / v to 0.1% w / v, and (d) 15 mM to 35 mM histidine buffer The drug formulation has a pH range of 4.5 to 5.5.

[0123] In some embodiments, an aqueous pharmaceutical preparation is disclosed, the aqueous pharmaceutical preparation comprising: (a) an isolated anti-Aβ primary fibrillation antibody or fragment thereof at concentrations ranging from 80 mg / mL to 240 mg / mL, the isolated anti-Aβ primary fibrillation antibody or fragment thereof comprising: (i) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:1, and (ii) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:2. (b) Arginine hydrochloride at 140 mM to 260 mM (c) Polysorbate 80, from 0.01% w / v to 0.1% w / v, and (d) 15 mM to 35 mM histidine buffer The drug formulation has a pH range of 4.5 to 5.5.

[0124] In some embodiments, an aqueous pharmaceutical preparation is disclosed, the aqueous pharmaceutical preparation comprising: (a) an isolated anti-Aβ primary fibrillation antibody or fragment thereof at concentrations from 80 mg / mL to 240 mg / mL, the isolated anti-Aβ primary fibrillation antibody or fragment thereof comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 11, and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 12. (b) Arginine hydrochloride at 140 mM to 260 mM (c) Polysorbate 80, from 0.01% w / v to 0.1% w / v, and (d) 15 mM to 35 mM histidine buffer The drug formulation has a pH range of 4.5 to 5.5.

[0125] In some embodiments, this document discloses an aqueous pharmaceutical preparation comprising: (a) BAN2401 at concentrations ranging from 80 mg / mL to 120 mg / mL, (b) Arginine at 240 mM to 360 mM, (c) Polysorbate 80, ranging from 0.03% w / v to 0.08% w / v, and (d) 30 mM to 70 mM citrate buffer The pharmaceutical preparation has a pH in the range of 4.5 to 5.5, and the arginine is arginine, arginine hydrochloride, or a combination thereof.

[0126] In some embodiments, this document discloses an aqueous pharmaceutical preparation comprising: (a) An isolated anti-Aβ primary fibrillation antibody or fragment thereof at concentrations of 80 mg / mL to 120 mg / mL, the isolated anti-Aβ primary fibrillation antibody or fragment thereof comprising: (i) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:1, and (ii) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:2. (b) Arginine at 240 mM to 360 mM, (c) Polysorbate 80, ranging from 0.03% w / v to 0.08% w / v, and (d) 30 mM to 70 mM citrate buffer The pharmaceutical preparation has a pH in the range of 4.5 to 5.5, and the arginine is arginine, arginine hydrochloride, or a combination thereof.

[0127] In some embodiments, this document discloses an aqueous pharmaceutical preparation comprising: (a) an isolated anti-Aβ primary fibrillation antibody or fragment thereof at a concentration of 80 mg / mL to 120 mg / mL, the isolated anti-Aβ primary fibrillation antibody or fragment thereof comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 11, and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 12. (b) Arginine at 240 mM to 360 mM, (c) Polysorbate 80, ranging from 0.03% w / v to 0.08% w / v, and (d) 30 mM to 70 mM citrate buffer The pharmaceutical preparation has a pH in the range of 4.5 to 5.5, and the arginine is arginine, arginine hydrochloride, or a combination thereof.

[0128] In some embodiments, this document discloses an aqueous pharmaceutical preparation comprising: (a) BAN2401 at 100 mg / mL (b) 200 mM arginine hydrochloride, (c) 0.05% w / v polysorbate 80, and (d) 25mM histidine buffer The drug formulation has a pH range of 4.5 to 5.5.

[0129] In some embodiments, this document discloses an aqueous pharmaceutical preparation comprising: (a) 100 mg / mL of isolated anti-Aβ primary fibrillation antibody or a fragment thereof, the isolated anti-Aβ primary fibrillation antibody or fragment thereof comprising: (i) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:1, and (ii) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:2. (b) 200 mM arginine hydrochloride, (c) 0.05% w / v polysorbate 80, and (d) 25mM histidine buffer The drug formulation has a pH range of 4.5 to 5.5.

[0130] In some embodiments, this document discloses an aqueous pharmaceutical preparation comprising: (a) 100 mg / mL of isolated anti-Aβ primary fibrillation antibody or a fragment thereof, the isolated anti-Aβ primary fibrillation antibody or fragment thereof comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO:11, and (ii) a light chain comprising the amino acid sequence of SEQ ID NO:12. (b) 200 mM arginine hydrochloride, (c) 0.05% w / v polysorbate 80, and (d) 25mM histidine buffer The drug formulation has a pH range of 4.5 to 5.5.

[0131] In some embodiments, this document discloses an aqueous pharmaceutical preparation comprising: (a) BAN2401 at 200 mg / mL (b) 200 mM arginine hydrochloride, (c) 0.05% w / v polysorbate 80, and (d) 25mM histidine buffer The drug formulation has a pH range of 4.5 to 5.5.

[0132] In some embodiments, this document discloses an aqueous pharmaceutical preparation comprising: (a) 200 mg / mL of isolated anti-Aβ primary fibrillation antibody or a fragment thereof, the isolated anti-Aβ primary fibrillation antibody or fragment thereof comprising: (i) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:1, and (ii) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:2. (b) 200 mM arginine hydrochloride, (c) 0.05% w / v polysorbate 80, and (d) 25mM histidine buffer The drug formulation has a pH range of 4.5 to 5.5.

[0133] In some embodiments, this document discloses an aqueous pharmaceutical preparation comprising: (a) 200 mg / mL of isolated anti-Aβ primary fibrillation antibody or a fragment thereof, the isolated anti-Aβ primary fibrillation antibody or fragment thereof comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO:11, and (ii) a light chain comprising the amino acid sequence of SEQ ID NO:12. (b) 200 mM arginine hydrochloride, (c) 0.05% w / v polysorbate 80, and (d) 25mM histidine buffer The drug formulation has a pH range of 4.5 to 5.5.

[0134] In some embodiments, this document discloses an aqueous pharmaceutical preparation comprising: (a) BAN2401 at 100 mg / mL (b) 350mM arginine, (c) 0.05% w / v polysorbate 80, and (d) 50 mM citrate buffer, The pharmaceutical preparation has a pH in the range of 4.5 to 5.5, and the arginine is arginine, arginine hydrochloride, or a combination thereof.

[0135] In some embodiments, this document discloses an aqueous pharmaceutical preparation comprising: (a) 100 mg / mL of isolated anti-Aβ primary fibrillation antibody or a fragment thereof, the isolated anti-Aβ primary fibrillation antibody or fragment thereof comprising: (i) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:1, and (ii) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:2. (b) 350mM arginine, (c) 0.05% w / v polysorbate 80, and (d) 50 mM citrate buffer, The pharmaceutical preparation has a pH in the range of 4.5 to 5.5, and the arginine is arginine, arginine hydrochloride, or a combination thereof.

[0136] In some embodiments, this document discloses an aqueous pharmaceutical preparation comprising: (a) 100 mg / mL of isolated anti-Aβ primary fibrillation antibody or a fragment thereof, the isolated anti-Aβ primary fibrillation antibody or fragment thereof comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO:11, and (ii) a light chain comprising the amino acid sequence of SEQ ID NO:12. (b) 350mM arginine, (c) 0.05% w / v polysorbate 80, and (d) 50 mM citrate buffer, The pharmaceutical preparation has a pH in the range of 4.5 to 5.5, and the arginine is arginine, arginine hydrochloride, or a combination thereof.

[0137] In some embodiments, this document discloses an aqueous pharmaceutical preparation comprising: (a) BAN2401 at concentrations of 150 mg / mL to 250 mg / mL, (b) 100mM to 150mM arginine hydrochloride (c) Polysorbate 80, ranging from 0.01% w / v to 0.05% w / v, and (d) 35 mM to 65 mM histidine buffer

[0138] The pharmaceutical formulation has a pH range of 4.5 to 5.5. In some embodiments, an aqueous pharmaceutical formulation is disclosed herein, comprising: (a) An isolated anti-Aβ primary fibrillation antibody or fragment thereof at concentrations of 150 mg / mL to 250 mg / mL, the isolated anti-Aβ primary fibrillation antibody or fragment thereof comprising: (i) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:1, and (ii) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:2. (b) 100mM to 150mM arginine hydrochloride (c) Polysorbate 80, ranging from 0.01% w / v to 0.05% w / v, and (d) 35 mM to 65 mM histidine buffer The drug formulation has a pH range of 4.5 to 5.5.

[0139] In some embodiments, this document discloses an aqueous pharmaceutical preparation comprising: (a) an isolated anti-Aβ primary fibrillation antibody or fragment thereof at concentrations of 150 mg / mL to 250 mg / mL, the isolated anti-Aβ primary fibrillation antibody or fragment thereof comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 11, and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 12. (b) 100mM to 150mM arginine hydrochloride (c) Polysorbate 80, ranging from 0.01% w / v to 0.05% w / v, and (d) 35 mM to 65 mM histidine buffer The drug formulation has a pH range of 4.5 to 5.5.

[0140] In some embodiments, the pH of the aqueous pharmaceutical preparation is 5.0. Methods to reduce aggregate formation

[0141] In some embodiments, this document discloses a method for reducing the formation of aggregates of isolated anti-Aβ protofibrillary antibodies or fragments thereof, the method comprising: (a) Providing a pharmaceutical preparation comprising, in a concentration of 50 mg / mL or higher of an isolated anti-Aβ primary fibrillary antibody or a fragment thereof that binds to human Aβ primary fibrillary tissue, optionally wherein the concentration is from 80 mg / mL to 300 mg / mL, optionally wherein the concentration is from 100 mg / mL to 200 mg / mL, and (b) Add arginine, arginine hydrochloride, or a combination thereof, and optionally provide a pharmaceutically acceptable buffer, wherein the buffer is optionally a histidine buffer, wherein the isolated anti-Aβ primary fibrillary antibody or fragment thereof comprises (i) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:1, and (ii) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:2.

[0142] In some embodiments of the method for reducing aggregate formation, the pH of the pharmaceutical formulation ranges from 4.5 to 5.5. In some embodiments of the method, the pH of the pharmaceutical formulation is 5.0.

[0143] In some embodiments of the method for reducing aggregate formation, the arginine, arginine hydrochloride, or a combination thereof is present at a concentration of 150 mM to 250 mM.

[0144] In some embodiments, this document discloses a method for reducing the fragmentation of isolated anti-Aβ protofibrillary antibodies or fragments thereof, the method comprising: (a) Providing a pharmaceutical preparation comprising, in a concentration of 50 mg / mL or higher of an isolated anti-Aβ primary fibrillary antibody or a fragment thereof that binds to human Aβ primary fibrillary tissue, optionally wherein the concentration is from 80 mg / mL to 300 mg / mL, optionally wherein the concentration is from 100 mg / mL to 200 mg / mL, and (b) Adding a histidine buffer and optionally further providing arginine, arginine hydrochloride, or a combination thereof, wherein the isolated anti-Aβ primary fibrillary antibody or fragment thereof comprises (i) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:1, and (ii) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:2.

[0145] In some embodiments of the method for reducing fragmentation, the pH of the pharmaceutical formulation ranges from 4.5 to 5.5. In some embodiments of the method, the pH of the pharmaceutical formulation is 5.0.

[0146] In some embodiments of the method for reducing fragmentation, the arginine, arginine hydrochloride, or a combination thereof is present at a concentration of 100 mM to 400 mM.

[0147] In some embodiments of the method for reducing fragmentation, the pharmaceutical formulation further comprises 0.01% w / v to 0.1% w / v of polysorbate 80.

[0148] In some embodiments of the method for reducing fragmentation, the pharmaceutical formulation further comprises a pharmaceutically acceptable buffer, wherein the buffer is a histidine buffer, optionally in the range of 10 mM to 100 mM, 12 mM to 80 mM, 14 mM to 60 mM, or 15 mM to 55 mM, 15 mM to 35 mM, or 15 mM to 25 mM.

[0149] In some embodiments, this document discloses a method for reducing aggregate formation and / or fragmentation of isolated anti-Aβ protofibrillary antibodies or fragments thereof, the method comprising: (a) Providing a pharmaceutical preparation comprising, in a concentration of 50 mg / mL or higher of isolated anti-Aβ primary fibrillary antibody or a fragment thereof that binds to human Aβ primary fibrillary cells, optionally wherein the concentration is from 80 mg / mL to 300 mg / mL, optionally wherein the concentration is from 100 mg / mL to 200 mg / mL. (b) Add arginine, arginine hydrochloride, or a combination thereof, and (c) Provide a pharmaceutically acceptable histidine buffer, wherein the isolated anti-Aβ primary fibrillary antibody or a fragment thereof comprises (i) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:1, and (ii) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:2.

[0150] In some embodiments, this document discloses a method for reducing aggregate formation and / or fragmentation of isolated anti-Aβ protofibrillary antibodies or fragments thereof, the method comprising: (a) Providing a pharmaceutical preparation comprising, in a concentration of 50 mg / mL or higher of isolated anti-Aβ primary fibrillary antibody or a fragment thereof that binds to human Aβ primary fibrillary cells, optionally wherein the concentration is from 80 mg / mL to 300 mg / mL, optionally wherein the concentration is from 100 mg / mL to 200 mg / mL. (b) Provide histidine buffer, and (c) Provide arginine, arginine hydrochloride, or a combination thereof, wherein the pH of the formulation is 4.5-5.5, wherein the isolated anti-Aβ primary fibrillary antibody or a fragment thereof comprises (i) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:1, and (ii) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:2.

[0151] This article also provides the following examples: [Example 01] An aqueous pharmaceutical preparation comprising: a. Isolated anti-Aβ primary fibrillation antibodies or fragments thereof that bind to human Aβ primary fibrillation fibers, at concentrations ranging from 80 mg / mL to 300 mg / mL. b. 100mM to 400mM of arginine, c. Polysorbate 80, from 0.01% w / v to 0.1% w / v, and d. Pharmaceutically acceptable buffers, The drug formulation has a pH range of 4.5 to 5.5. The isolated anti-Aβ primary protoplasmic antibody or its fragment comprises: (i) a heavy chain variable domain containing the amino acid sequence of SEQ ID NO:1, and (ii) a light chain variable domain containing the amino acid sequence of SEQ ID NO:2. The arginine in question is arginine, arginine hydrochloride, or a combination thereof. [Example 02] The pharmaceutical formulation according to claim 1, wherein the isolated anti-Aβ primary fibrillary antibody or a fragment thereof is present at a concentration of 100 mg / mL to 200 mg / mL. [Example 03] The pharmaceutical preparation according to claim 1, wherein the isolated anti-Aβ primary fibrillary antibody or a fragment thereof is present at a concentration of 100 mg / mL. [Example 04] The pharmaceutical formulation according to claim 1, wherein the isolated anti-Aβ primary fibrillary antibody or a fragment thereof is present at a concentration of 200 mg / mL. [Example 05] The pharmaceutical formulation according to any one of claims 1-4, wherein the pharmaceutical formulation further comprises methionine. [Example 06] A pharmaceutical formulation according to any one of claims 1-5, wherein the pharmaceutically acceptable buffer is a citrate buffer or a histidine buffer. [Example 07] A pharmaceutical formulation according to any one of claims 1-6, wherein the pharmaceutical formulation comprises 10 to 100 mM of citrate buffer or 10 to 100 mM of histidine buffer. [Example 08] A pharmaceutical preparation according to any one of claims 1-7, wherein the pharmaceutical preparation comprises 125 to 350 mM of arginine. [Example 09] The pharmaceutical preparation according to any one of claims 1-8, wherein the pharmaceutical preparation contains 200 mM of arginine, wherein the arginine is arginine hydrochloride. [Example 10] A pharmaceutical preparation according to any one of claims 1-9, the pharmaceutical preparation comprising 200 mM arginine and 25 mM histidine buffer, wherein the arginine is arginine hydrochloride. [Example 11] An aqueous pharmaceutical preparation comprising: a. An isolated anti-Aβ primary fibrillation antibody or fragment thereof at concentrations ranging from 80 mg / mL to 240 mg / mL, the isolated anti-Aβ primary fibrillation antibody or fragment thereof comprising: (i) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:1, and (ii) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:2. b. Arginine hydrochloride ranging from 140 mM to 260 mM c. Polysorbate 80, ranging from 0.02% w / v to 0.08% w / v, and d. 15mM to 35mM histidine buffer, The drug formulation has a pH range of 4.5 to 5.5. [Example 12] An aqueous pharmaceutical preparation comprising: a. An isolated anti-Aβ primary fibrillation antibody or fragment thereof at concentrations from 80 mg / mL to 120 mg / mL, the isolated anti-Aβ primary fibrillation antibody or fragment thereof comprising: (i) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:1, and (ii) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:2. b. Arginine from 240mM to 360mM c. Polysorbate 80, ranging from 0.02% w / v to 0.08% w / v, and d. 30mM to 50mM citrate buffer, The drug formulation has a pH range of 4.5 to 5.5. And the arginine in it is arginine, arginine hydrochloride, or a combination thereof. [Example 13] A method for reducing the formation of aggregates of isolated anti-Aβ primary proficiency antibodies or fragments thereof, the method comprising: An aqueous pharmaceutical formulation comprising an isolated anti-Aβ primary fibrillation antibody or a fragment thereof, the isolated anti-Aβ primary fibrillation antibody or fragment thereof comprising: (i) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:1, and (ii) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:2, wherein the isolated anti-Aβ primary fibrillation antibody or fragment thereof is present at a concentration of 80 mg / ml to 300 mg / ml, and Arginine, arginine hydrochloride, or a combination thereof are added to the aqueous pharmaceutical preparation, wherein the pH range of the pharmaceutical preparation is from 4.5 to 5.5. [Example 14] The method according to claim 13, wherein the arginine, arginine hydrochloride, or combination thereof is present at a concentration of 150 to 250 mM. [Example 15] The method according to claim 14, wherein the arginine, arginine hydrochloride, or combination thereof is present at a concentration of 200 mM. [Example 16] The method according to any one of claims 13 to 15, wherein the pharmaceutical preparation further comprises a pharmaceutically acceptable buffer. [Example 17] The method according to claim 16, wherein the pharmaceutically acceptable buffer is a histidine buffer or a citrate buffer. [Example 18] A method for reducing the fragmentation of isolated anti-Aβ primary fibrillary antibodies or fragments thereof, the method comprising: An aqueous pharmaceutical formulation comprising an isolated anti-Aβ primary fibrillation antibody or a fragment thereof, the isolated anti-Aβ primary fibrillation antibody or fragment thereof comprising: (i) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:1, and (ii) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:2, wherein the isolated anti-Aβ primary fibrillation antibody or fragment thereof is present at a concentration of 80 mg / ml to 300 mg / ml, and Histidine buffer is added to the aqueous pharmaceutical composition, wherein the pH range of the pharmaceutical formulation is from 4.5 to 5.5. [Example 19] The method according to claim 18, wherein the method comprises 15 to 35 mM of histidine buffer. [Example 20] The method according to any one of claims 13-19, wherein the pharmaceutical formulation further comprises 0.01% w / v to 0.1% w / v of polysorbate 80. [Example 21] An aqueous pharmaceutical preparation comprising: a. An isolated anti-Aβ primary fibrillary antibody or fragment thereof at concentrations from 80 mg / mL to 240 mg / mL, the isolated anti-Aβ primary fibrillary antibody or fragment thereof comprising: (i) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:1, and (ii) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:2; b. Arginine, arginine hydrochloride, or a combination thereof, in the range of 140 mM to 260 mM; c. Polysorbate 80, ranging from 0.01% w / v to 0.1% w / v; and d. 15 mM to 35 mM histidine buffer or 30 mM to 50 mM citrate buffer. The drug formulation has a pH range of 4.5 to 5.5. [Example 22] The pharmaceutical preparation according to Example 21 contains 100 mg / mL of the antibody. [Example 23] The pharmaceutical preparation according to Example 21 contains 200 mg / mL of the antibody. [Example 24] The pharmaceutical preparation according to Example 21 contains the antibody at concentrations of 90 mg / mL to 220 mg / mL, 100 mg / mL to 210 mg / mL, or 110 mg / mL to 200 mg / mL. [Example 25] A pharmaceutical preparation according to any one of Examples 21-24, the pharmaceutical preparation comprising a histidine buffer, wherein the histidine buffer comprises 15 mM to 30 mM, 15 mM to 25 mM, or 20 mM to 30 mM of histidine. [Example 26] A pharmaceutical formulation according to any one of Examples 21-25, the pharmaceutical formulation comprising a histidine buffer, wherein the histidine buffer comprises 0.2-0.3 mg / mL of histidine and 4.4 to 4.9 mg / mL of histidine hydrochloride, optionally the histidine hydrochloride being a monohydrate. [Example 27] A pharmaceutical formulation according to any one of Examples 21-26, the pharmaceutical formulation comprising 150 mM to 250 mM, 160 mM to 240 mM, 170 mM to 230 mM, 180 mM to 220 mM, 190 mM to 210 mM of arginine, arginine hydrochloride, or a combination thereof. [Example 28] A pharmaceutical formulation according to any one of Examples 21-27, the pharmaceutical formulation comprising 0.02% w / v to 0.05% or 0.04% w / v to 0.04% w / v of polysorbate 80. [Example 29] A pharmaceutical preparation according to any one of Examples 21-28, the pharmaceutical preparation comprising a citrate buffer. [Example 30] A pharmaceutical preparation according to any one of Examples 21-29, wherein the pharmaceutical preparation comprises a histidine buffer. [Example 31] The pharmaceutical formulation according to any one of Examples 21-30 further comprises methionine. [Example 32] An aqueous pharmaceutical preparation comprising: a. An antibody of 100 mg / ml to 200 mg / ml, comprising: (i) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:1, and (ii) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:2; b. 200 mM arginine hydrochloride; c. 0.05% w / v polysorbate 80; and d. 25mM histidine buffer, The drug formulation has a pH range of 4.5 to 5.5. [Example 33] The pharmaceutical preparation according to Example 32 contains 100 mg / mL of the antibody. [Example 34] The pharmaceutical preparation according to Example 32 contains 200 mg / mL of the antibody. [Example 35] A method for reducing antibody aggregate formation and / or fragmentation, the method comprising: a. Provides an aqueous pharmaceutical formulation comprising an isolated anti-Aβ primary fibrillation antibody or a fragment thereof, the isolated anti-Aβ primary fibrillation antibody or fragment thereof comprising: (i) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:1, and (ii) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:2, wherein the isolated anti-Aβ primary fibrillation antibody or fragment thereof is present at a concentration of 80 mg / ml to 300 mg / ml, and b. Add 1) arginine, arginine hydrochloride, or a combination thereof, or 2) histidine buffer to the aqueous pharmaceutical composition. [Example 36] The method according to Example 35, wherein the pH range of the pharmaceutical preparation is from 4.5 to 5.5. [Example 37] The method according to Example 35, wherein the arginine, arginine hydrochloride, or combination thereof is present at a concentration of 150-250 mM. [Example 38] The method according to Example 37, wherein the arginine, arginine hydrochloride, or a combination thereof is present at a concentration of 200 mM. [Example 39] The method according to Example 38, wherein the method comprises 15 to 25 mM of histidine buffer. [Example 40] The method according to any one of Examples 35-39, wherein the pharmaceutical formulation further comprises 0.01% w / v to 0.1% w / v of polysorbate 80. [Example 41] The method according to any one of Examples 35-40, wherein the pharmaceutical preparation further comprises a pharmaceutically acceptable buffer. [Example 42] The method according to Example 41, wherein the pharmaceutically acceptable buffer is a histidine buffer or a citrate buffer. Example Example 1: Protein Concentration Study

[0152] Samples with three protein concentrations (10, 100, and 200 mg / mL) were prepared and tested in concentration studies. The 10 mg / mL material was pure BAN2401 drug substance (PDS), while the 100 and 200 mg / mL materials were produced from BAN2401 PDS using an Amicon Ultra-15 rotary filter. All samples were prepared in the same formulation buffer (25 mM sodium citrate, 125 mM sodium chloride, pH 5.7), but each sample had a different PS80 level. The percentages of PS80 in the 10, 100, and 200 mg / mL samples were 0.02%, 0.16%, and 0.32%, respectively. All samples were filtered through a 0.2 gm filter and aliquoted into sterile polypropylene (PP) tubes for stability testing. (Note: The PS80 removal process was not available for concentration studies.)

[0153] Sample stability was assessed over 3 months using non-denaturing HPLC-SEC, pH, and DLS at two temperatures (5°C and 25°C). Additional characterization assays, such as PS80 assay, were performed on samples at T=0. Table 1 shows detailed information for the tested samples. Table 1. Testing Schedule (Concentration Studies)

[0154] pH was measured using a pH meter with a microprobe. Calibration was performed using pH 4.0 and 7.0 standard solutions prior to measurement.

[0155] The aggregation and physical degradation of concentrated BAN2401 were evaluated by non-denaturing HPLC-SEC of stable samples stored at 5°C and 25°C.

[0156] HPLC-SEC (non-denaturing) analysis was performed using a TSK G3000 SWXL column (mobile phase: 0.2 M sodium phosphate, pH 7.0, flow rate: 1.0 mL / min). For protein concentrations of 10, 100, or 200 mg / mL, sample injection volumes were 15, 1.5, or 0.8 pg, respectively. This ensured that approximately 150 pg of protein was injected onto the column throughout the study. Results showing the relative peak areas (%) of monomers, aggregates, and fragments are reported in [link to report]. Figure 1-6 middle.

[0157] At both temperatures (5°C and 25°C), both the percentage and rate of aggregation increased with increasing protein concentration. For the 100 and 200 mg / mL samples, the initial percentage of aggregation was almost twice that of the 10 mg / mL sample. This indicates that the concentration process alone leads to protein aggregation. Additional aggregation occurred during storage at 5°C and 25°C. For 100 mg / mL, the average aggregation rate was 0.17% per month at 5°C for 3 months, and 0.20% per month for 200 mg / mL. For comparison, the 10 mg / mL sample showed an aggregation rate of only 0.07% per month under the same conditions. In conclusion, BAN2401 at concentrations of 100 mg / mL or higher appears to be physically unstable in formulations.

[0158] Protein concentration was assessed by measuring absorbance at 280 and 320 μm using a Beckman DU-800 spectrophotometer with UV light. Samples were diluted 500-fold and prepared in duplicate. Protein concentration was calculated using the following formula with an extinction coefficient ε of 1.32: Concentration = (A280 - A320) / εx Dilution factor

[0159] Protein concentrations were measured using a UV-Vis spectrophotometer. The results are listed in Table 2. Overall, the concentrations remained constant throughout the three months at both temperatures. Table 2. Protein Concentration Results (Concentration Study) Example 2: pH Screening Study

[0160] The stability of 200 mg / mL BAN2401 was evaluated at five different pH values. To prepare samples, PS80-free BAN2401 PDS was concentrated and percolated into a buffer (pH 4.5) containing 50 mM sodium citrate and 100 mM sodium chloride. Final concentration adjustments were made via dilution to achieve a concentration of 200 mg / mL protein and 0.02% PS80. The resulting material was aliquoted into five equal aliquots; four of the aliquots were titrated with 10 N sodium hydroxide to produce samples at different pH values ​​(i.e., pH 5.0, 5.5, 6.0, and 6.5). The resulting samples were filtered through a 0.2 μm filter and then aliquoted into sterile polypropylene (PP) tubes for stability assessment.

[0161] Sample stability was assessed over a 3-month period at two temperatures (5°C and 25°C) using non-denaturing HPLC-SEC, pH, and DLS. Additionally, a stirring study was conducted at the 1-month timepoint, in which a subset of the 5°C samples was stirred at 250 RPM for 3 days in a 25°C incubator. Table 3 shows detailed information for the tested samples. Table 3. Testing Schedule (pH Study)

[0162] SEC-HPLC analysis was performed using a TSK G3000 SWXL column (mobile phase: 0.2 M sodium phosphate, pH 7.0, flow rate: 1.0 mL / min). Sample injection volumes were 15, 1.5, or 0.8 μL for protein concentrations of 10, 100, or 200 mg / mL, respectively. This ensured that approximately 150 μg of protein was injected onto the column throughout the study. The results for the relative peak areas (%) of monomers, aggregates, and fragments are reported.

[0163] The aggregation and physical degradation of concentrated BAN2401 were evaluated by non-denaturing HPLC-SEC of stable samples stored at 5°C and 25°C. Results are shown in... Figure 7-12 At lower pH values, BAN2401 appeared to be more stable at both temperatures (5°C and 25°C). Formulations at pH 4.5 and 5.0 showed lower initial aggregate percentages and exhibited slower aggregate growth, with a rate of approximately 0.07% per month at 5°C. In addition to aggregation, protein fragmentation was also analyzed. The results indicated that BAN2401 fragmentation was more pronounced in BAN2401 formulations at lower pH values. Based on these considerations, the BAN2401 formulation at pH 5.0 was considered the most stable (i.e., exhibiting less aggregation and / or physical degradation of BAN2401 compared to other formulations at different pH values). Example 3: Excipient Screening Study

[0164] Twelve excipients were screened in this study. To prepare samples containing 200 mg / mL BAN2401, BAN2401 with PS80 removed was concentrated via a TFF step followed by a percolation step with a base buffer (50 mM citrate, 0.02% PS80, pH 6.0). The concentrated material was divided into twenty-four fractions. Each fraction was infused with a stock solution containing the specific excipient. For most excipients, two concentrations were examined, except for sodium chloride and ascorbic acid. For sodium chloride, samples were prepared at three concentration levels; for ascorbic acid, samples were prepared at one concentration level. Table 4 shows a list of the excipients used and their concentrations. Table 4. List of Excipients and Corresponding Concentrations

[0165] All samples, including the control (i.e., samples in a base buffer without excipients), were filtered to 0.2 gm and then aseptically filled into BD syringes using a manual stopper. Control samples were also filled into glass vials. The vials and BD syringes were stored at 5°C and 25°C, respectively. Sample stability was assessed for 2 months using non-denaturing HPLC-SEC and pH.

[0166] Samples for the excipient screening study were prepared under suboptimal buffer conditions (50 mM citrate, 0.02% PS80, pH 6.0) to expand the positive role of excipients in preventing aggregate formation. The stability characteristics (aggregation) of each formulation at 25°C for 2 months were evaluated by non-denaturing HPLC-SEC.

[0167] like Figure 13-15 As shown, among these formulations, the formulation containing 160 mM arginine (F14) produced the lowest aggregation rate. For comparison, the 160 mM arginine formulation showed only 1.4% aggregates after 2 months at 25°C, while the control (i.e., the formulation in a base buffer without excipients—F0) showed 2.4% aggregates at the same time under the same conditions.

[0168] Furthermore, the formulation containing arginine exhibited the lowest initial aggregation percentage, indicating that arginine inhibits aggregate formation during the protein concentration step. The arginine formulation (F14) showed a slightly higher degree of fragmentation compared to the control (F0). However, this difference is likely within the scope of assay variability.

[0169] The second best excipient in this study was the formulation with 400 mM proline (F10), although it was not as effective as the arginine formulation (F14) in controlling aggregation.

[0170] Compared to the control samples, the formulations containing sodium chloride (F16-F18) had no effect on stability. The same observation was made for the formulations containing PS80 (F19 and F20): no stability effect was observed by changing the PS80 content. The formulation containing ascorbic acid (F15) showed a significant effect of increasing the percentage of aggregates and fragments from the start of the study.

[0171] The pH of two arginine formulations (F13 and F14) was measured to verify that the presence of arginine in the formulations did not cause pH changes over time. As shown in Table 4, no pH changes were observed. The osmolality of the two formulations was also measured. These samples were stored at -20°C before being thawed simultaneously for measurement. Table 5. pH and osmolality of arginine formulations (F13 and F14) Example 4: Arginine Concentration Study

[0172] The relationship between arginine concentration and aggregates in the formulation is provided by Design 7.0 Conduct the research. Table 6 below shows the factors selected for the research, as well as the level of exploration for each factor. Table 6. Factors studied in Example 4

[0173] Through D-optimal design in Design Sample randomization was initiated in section 7.0. The resulting sample set contained 25 data points, including 4 replicates, as shown in Table 7. Each formulation was prepared and evaluated. Table 7. Formulations prepared for Example 4

[0174] Through Design 7.0 The relationship between arginine concentration and aggregate levels in a 100 mg / mL BAN2401 formulation (pH 5.0) containing 0.02% polysorbate 80 was evaluated. The estimated results are shown in... Figure 16 middle. Example 5: PS80 Research

[0175] To assess changes in drug product quality under stress conditions, agitation and freeze-thaw studies were conducted using formulation buffers (pH 5.0, 350 mM arginine, 50 mM citric acid). In this experiment, formulations with varying polysorbate 80 concentrations were investigated to confirm the effect of polysorbate 80 on subvisible particle formation under such stress conditions. Table 8 shows the formulations studied and the applied stress conditions. Table 8. Studies on formulations and freeze-thaw stresses investigated by stirring

[0176] Samples were prepared in the same manner as described in Example 4. 10% PS80 solution was added to achieve the target PS80 concentration in the formulation, and the final protein concentration was adjusted by dilution with formulation buffer. The formulation was passed through a 0.2 gm filter and filled into 2 mL vials at a filling volume of 1.3 mL. The samples were placed horizontally (via the vials sideways) on a track-mounted shaker and then placed in a freezer or on a lab bench. The samples were shaken at 250 rpm for 3 days. Other samples were frozen for 2 hours at -20°C, then removed and allowed to thaw at room temperature for 2 hours. This freeze-thaw cycle was repeated three times. The visual appearance, aggregate and fragment levels of the samples from the agitation and freeze-thaw studies were evaluated by SEC-HPLC, and sub-visible particles were evaluated by microflow imaging (MFI).

[0177] The following summarizes the results of agitation and freeze-thaw studies on BAN2401 formulations containing different levels of PS80. SEC-HPLC analysis

[0178] SEC-HPLC analysis was performed using a TSK G3000 SWXL column (mobile phase: 0.2 M sodium phosphate, pH 7.0, flow rate: 1.0 mL / min). For protein concentrations of 50 or 100 mg / mL, sample injection volumes were 5.0 μL or 2.5 μL, respectively. This ensured that approximately 250 μg of protein was injected onto the column throughout the study. The results for the relative peak areas (%) of monomers, aggregates, and fragments are reported.

[0179] Regarding aggregate levels in formulations with varying polysorbate 80 concentrations and stored at 25°C for 3 months at pH 5, there was no significant difference in aggregate levels when the PS80 concentration increased from 0% to 0.06% and the formulation pH was 5.0. It can be concluded that PS80 does not affect aggregate formation (i.e., dimers and trimers as measured by SEC-HPLC) during storage of BAN2401 formulations at 25°C.

[0180] Regarding the effect of polysorbate 80 concentration on fragment levels in formulations at pH 5, polysorbate 80 concentration did not affect fragment levels in BAN2401.

[0181] Regarding the appearance of the preparations after 3 days of agitation at 250 rpm at ambient temperature, the sample without PS80 was cloudy with precipitated protein, while the other samples (0.02%, 0.05%, and 0.1% polysorbate 80) were clearly clear, particle-free solutions. All samples agitated at 250 rpm at 5°C for 3 days were clear and free of precipitate. All samples subjected to freeze / thaw cycles were also clear and free of precipitate.

[0182] Table 9 shows the aggregate and fragment levels in BAN2401 formulations subjected to agitation and freeze-thaw cycles. The stress condition causing the greatest instability was agitation under ambient conditions. Under these conditions, the PS80-free sample exhibited the most high molecular weight formation and the greatest monomer loss. This effect diminished with increasing PS80 concentration. Table 9. Results at the aggregate and fragment levels from the agitation and freeze-thaw studies. (a) High molecular weight substances; (b) Low molecular weight substances; and (c) The sample is turbid. Subvisible particle analysis

[0183] Sub-visible particle analysis was performed using microfluidic imaging (DPA4100 flow imaging microscope and BP-4100-FC-400-UN flow cell). The total sample volume was 0.9 mL, and results are reported for the 2.25–100 μm, 5–100 μm, 10–100 μm, and 25–100 μm ranges.

[0184] An increase in subvisible particles (2 to 10 pm) was observed in PS80-free samples subjected to vibration and freeze-thaw cycles. The formation of subvisible particles was increasingly suppressed with increasing PS80 concentration in the formulation. This effect was observed across all size ranges studied. No significant changes in subvisible particles were observed in formulations containing 0.05% and 0.1% polysorbate 80. Therefore, a PS80 concentration of 0.05% was chosen as the optimal concentration for the formulation.

[0185] Data indicate that, as measured by SEC-HPLC, PS80 has no effect on the formation of BAN2401 aggregates (dimers and trimers) and fragments. Therefore, the PS80 concentration was selected based on the results of agitation and freeze-thaw studies. A PS80 concentration of 0.05% was chosen to prevent possible precipitation during transport and to minimize the formation of subvisible particles. Example 6: Preparation of intravenous (IV) preparations of BAN2401 A.BAN2401 10mg / mL preparation

[0186] BAN2401 10 mg / mL preparations (“10 mg / mL injections”) for intravenous injection were manufactured using standard cGMP aseptic methods for preparing sterile aqueous preparations. BAN2401 10 mg / mL injections were produced from the corresponding BAN2401 active pharmaceutical ingredient formulations (without the addition of any excipients or diluents) listed below. Table 10 shows exemplary IV preparations containing 10 mg / mL BAN2401. Table 10. Comparative 10 mg / mL IV formulations containing BAN2401. BAN2401 10mg / mL Sodium citrate / citric acid buffer 25mM Sodium chloride 125mM Polysorbate 80 0.02% (w / v) Water for Injection QS pH 5.7

[0187] like Figure 17 As shown, the filtered BAN2401 active pharmaceutical ingredient solution was aseptically filled into vials (in... Figure 17 (Referring to "Formulation A" in Chinese). The combined active pharmaceutical ingredient (API) undergoes a bioburden reduction filtration step through a 0.2-pm filter. Final aseptic filtration is performed through two 0.2-pm filters in series, with filter integrity tests conducted before and after filtration. The sterile bulk drug product is aseptically filled into vials. During filling, the filling accuracy is confirmed by measuring the vial filling weight. The filled vials are plugged and then sealed with an aluminum top seal. After crimping the cap, the product is stored at 5°C ± 3°C.

[0188] The vials were analyzed according to USP 788 using Method 1 (light obscuration method). The results are shown in Tables 11 and 12. Table 11. Particle size distribution of 10 mg / mL injection vials (number of particles / vial) Table 12. Particle size distribution of 10 mg / mL injection vials (particles / mL) Example 7: Preparation of intravenous (IV) and subcutaneous (SC) formulations of BAN2401 A.BAN2401 100mg / mL preparation

[0189] BAN2401 100 mg / mL formulation (“100 mg / mL injection”) is manufactured using standard cGMP aseptic methods for preparing sterile aqueous formulations. BAN2401 100 mg / mL is produced from the corresponding BAN2401 active pharmaceutical ingredient (without any excipients or diluents) (Table 13). Table 13. Exemplary 100 mg / mL IV preparations containing BAN2401.

[0190] like Figure 18 As shown, the filtered BAN2401 active pharmaceutical ingredient (100 mg / mL injection) solution was aseptically filled into vials (in... Figure 18 (Referring to "Formulation B" in Chinese). The active pharmaceutical ingredient (API) undergoes a bioburden reduction filtration step through a 0.2-pm filter. Final aseptic filtration is performed through two 0.2-pm filters in series, with filter integrity tests conducted before and after filtration. The sterile bulk drug product is aseptically filled into vials. Filling accuracy is confirmed by measuring the weight of the vials during filling. The filled vials are plugged and then sealed with an aluminum top seal. After crimping the cap, the product is stored at 5°C ± 3°C.

[0191] The vials were analyzed according to USP 788 using Method 1 (light obscuration method). The results are shown in Tables 14 and 15. Table 14. Particle size distribution of 100 mg / mL injection vials (number of particles / vial) 25μm 0 0 1 0 10μm 53 8 9 9 5μm 631 72 51 60 2μm 1954 321 249 267 Table 15. Particle size distribution of 100 mg / mL injection vials (particles / mL) 25μm 0 0 0.2 0 10μm 10.6 1.6 1.8 1.8 5μm 126.2 14.4 10.2 12 2μm 390.8 64.2 49.8 53.4

[0192] To manufacture another BAN2401 100 mg / mL injection. As shown in Table 16, the following materials may be used in a second exemplary formulation containing 100 mg / mL BAN2401. Table 16. Another exemplary 100 mg / mL IV preparation containing BAN2401. B.BAN2401 200mg / mL preparation

[0193] As shown in Table 17, the following materials can be used in exemplary SC formulations containing 200 mg / mL BAN2401. The stability of BAN2401 in these formulations (FSC1, FSC2, and FSC3) was evaluated in conjunction with material stability assessments of the following three container seals: i) Becton-Dickinson Hypak Biotech glass prefilled syringe (PFS) and stopper ii) West Crystal Zenith plastic PFS and stoppers iii) West 5mL glass vial and stopper Table 17. Exemplary 200 mg / mL SC formulations containing BAN2401. 1 Total concentration of histidine

[0194] BAN2401, with a target protein concentration of 200 mg / mL, was prepared via TFF as described below. Except for FSC1a and FSC1b, a separate TFF operation was performed on each formulation buffer to prepare the BAN2401 material. For both formulations, one TFF operation was performed, and the resulting concentrated material was divided into two half-lots. A small amount of sterile filter material from each final formulation buffer was not filled at zero temperature but was frozen at -20°C for injection testing into appropriate container seals. (a) Preparation of BAN2401

[0195] The protein concentration / percolation process via TFF can be divided into three stages: 1. Concentrate the material to 100-150 mg / mL 2. Percolation of the buffer in the formulation (5X) 3. Concentrate to >200mg / mL

[0196] The setting is 0.02m. 2 The concentration / percolation steps were performed using a Pall Centramate LV system. BAN2401 material (removed from GMP batch manufacturing prior to the addition of PS80) was loaded into the TFF system and concentrated 10–15 times (Stage 1). The material was then percolated against a formulation buffer at up to 5 times the percolation volume (Stage 2), with percolation monitored by pH and conductivity checks of the percolate. Following percolation, the material was further concentrated to a target protein concentration of 210–250 mg / mL (Stage 3). The percolate was collected and sampled to determine the protein concentration.

[0197] During the preparation of this formulation, the target protein concentration of 210 to 250 mg / mL was not achieved due to the high pressure in the TFF system. Therefore, the target protein concentration was achieved by using a Millipore 1-gal centrifuge and filtration unit (30,000 MWCO). To perform this concentration step, the filtration unit was equilibrated using BAN2401 formulation buffer, and then the BAN2401 material was centrifuged at 3600 RPM (approximately 3000 x g) for 30 minutes at 20°C until the expected protein concentration in the residue reached greater than 200 mg / mL. The residue was recovered from the filtration unit and combined. After thorough mixing, the combined residue was sampled for protein concentration measurement.

[0198] After protein concentration, the sample was removed from the pool and diluted 500-fold with an appropriate formulation buffer. The absorbance of the diluted sample at 280 nm and 320 nm was measured relative to the blank buffer. The final protein concentration was adjusted by dilution with an appropriate formulation buffer. Finally, 10% PS80 solution was added to the BAN2401 to bring the final solution to 0.02% PS80, and the protein solution was thoroughly mixed by inverted rotation.

[0199] The final BAN2401 preparation material was filtered using a 0.2 μm syringe filter and then filled into vials or PFS. This step was performed aseptically in a biosafety cabinet. The resulting vials or PFS were placed in a -20°C freezer. Vials were stored upside down, and PFS were stored horizontally to simulate worst-case conditions. C. Stability tests on an exemplary 200 mg / mL SC formulation containing BAN2401.

[0200] Use pH-specific ( Figure 19-21 ), absorbance at 405nm ( Figure 22-24 (to detect an increase in yellowing), size exclusion chromatography (SEC) ( Figure 25-33 The stability of the samples was evaluated at 5°C (3M, 6M, 9M, 12M) and 25°C (1M, 3M), and then at 5°C thereafter. Exemplary formulations FSC1-FSC4 are shown in Table 17; note that in... Figure 19-33In this context, "F1a" refers to FSC1a, "F1b" to FSC1b, "F2" to FSC2, and "F3" to FSC3. Additional characterization measurements were performed on samples at T=0, including protein concentration, differential scanning calorimetry (DSC), PS80, and osmolality. Sub-visible particle testing was conducted on samples at T=0 and T=6 months. A 3-day agitation study was performed on samples stored at 5°C for 1 month. Samples used for agitation studies were transferred to 25°C after 1 month of storage at 5°C and agitated at 250 rpm for 3 days.

[0201] Test results showed that 200 mg / mL BAN2401 in all FSC1a-FSC3 formulations exhibited similar aggregate and fragment levels after 12 months of storage at 2°C–8°C. The stability of 200 mg / mL BAN2401 in the three FSC1a-FSC3 formulations was evaluated. Overall, BAN2401 showed similar stability in each tested formulation, and after 12 months of storage at 5°C, it retained more than 98.2% of the monomers regardless of the container seal. Furthermore, after 12 months of storage at 5°C, the pH remained stable, and there was no significant increase in yellowing as detected by A405. (a) Protein concentration (sample at T=0)

[0202] Protein concentrations were measured using a UV-Vis spectrophotometer. The results are listed in Table 18. Table 18. Protein Concentration Results FSC1a 206 FSC1b 204 FSC2 201 FSC3 189

[0203] Protein concentration was assessed by measuring absorbance at 280 and 320 nm using a Beckman DU-800 spectrophotometer with a 1 cm quartz cuvette. Samples were diluted 500-fold and prepared in triplicate. Protein concentration was calculated using the following formula with an extinction coefficient ε of 1.32: Concentration = (A280 - A320) / εx Dilution factor (b) Polysorbate 80 (T=0 sample)

[0204] The PS80 content of the samples was measured by oleic acid quantification. The results are shown in Table 19. Table 19. Results of Polysorbate 80 FSC1a 0.016 FSC1b 0.022 FSC2 0.019 FSC3 0.019

[0205] The determination was performed by quantitative analysis of oleic acid (a hydrolysis product of PS80). Using alkaline hydrolysis, PS80 released oleic acid in a 1:1 molar ratio. Oleic acid was then separated from other PS80 hydrolysis products and the matrix using reversed-phase HPLC. Underrivatized oleic acid was monitored using absorbance at 195 nm. [J. Chromatography B, 878(2010)1865-1870]. Experimentally, the sample was mixed with sodium hydroxide to release oleic acid, followed by extraction with acetonitrile. The extract was diluted with potassium phosphate solution, and 100 μL of the sample was injected into a Waters Symmetry C18 column. Separation was performed using an isocratic eluent containing 80% organic phase A (acetonitrile) and 20% aqueous phase B (20 mM potassium dihydrogen phosphate, pH 2.8). The concentration of PS80 in the sample was calculated from the peak area using a standard curve. (c) Osmotic molar concentration (sample at T=0)

[0206] The osmolality of the samples was determined using a freezing point osmoremeter. The results are shown in Table 20. Table 20. Results of Osmotic Molar Concentration

[0207] Osmotic molality was measured using a Precision Systems Osmette III freezing-point osmoremeter. The sample was diluted 3-fold with WFI, and then 10 μL of the diluted sample was taken and loaded onto the instrument using the osmoremeter pipette. The resulting osmotic molality measurement was calibrated for further sample dilution. (d) Subvisible particles

[0208] Subvisible particle analysis was performed using a Fluid Technologies FlowCarn instrument. The results at T=0 are listed in Table 21. When the material was stored at 5°C, the total particle concentration and the number of particles larger than 10 μm appeared to increase over time. The magnitude of the increase is listed in the following order: FSC1a <FSC1b<FSC2<FSC3。 Table 21. Results of subvisible particles

[0209] Sub-visible particle analysis was performed using a FlowCam from Fluid Imaging Technologies, equipped with a 20x objective lens and a 50µm flow cell. Before running the BAN2401 sample, the flow cell was flushed with dI H2O, and the dI H2O was measured to ensure the flow cell was clean. The flow cell was considered usable if the total number of counted particles per 0.2 mL of dI H2O was ≤2. The sample was equilibrated to room temperature and then diluted 20-fold with deionized water. Repeated diluted samples were analyzed using a sample volume of 0.2 mL and a sample flow rate of 0.02 mL / min. The auto-imaging rate was 14, the efficiency was 19.6%, and the run time was 10 min. The resulting particle concentration was corrected for sample dilution. (e)pH

[0210] Throughout the stability testing process, the pH value of each formulation was monitored. For example... Figure 19-21 As shown, no significant changes were observed in the samples under any of the storage conditions tested. (f)HPLC-SEC

[0211] High-performance liquid chromatography-size exclusion chromatography (HPLC-SEC) was performed using a TSK 03000SWXL column (mobile phase: 0.2 M sodium phosphate, pH 7.0, flow rate: 1.0 mL / min). For a protein concentration of 200 mg / mL, the sample injection volume was 0.8 μL. This ensured that approximately 150 μg of protein was injected onto the column throughout the study. The results for the relative peak areas (%) of monomers, aggregates, and fragments are reported, as follows: Figure 25-33 As shown.

[0212] The physical degradation of BAN2401 was evaluated by HPLC-SEC of stability samples stored at 5°C and 25°C. After 12 months of storage at 5°C, the percentage of aggregates in the three formulations was similar, approximately 1.1%. The percentage of fragments generated in all tested formulations ranged from 0.4% to 0.5%. For all tested formulations and container seals, the monomer content ranged from 98.4% to 98.6% after 12 months of storage at 5°C. At the current degradation rate, after up to 24 months of storage at 5°C, BAN2401 in all tested formulations would likely contain more than 97% monomers.

[0213] After storage at 25°C for up to 3 months, the fragment content after 3 months ranged from 0.4% to 0.8%. Except for FSC3, all formulations had a monomer content slightly higher than 98%, while FSC3 had a monomer content slightly lower than 98% after storage at 25°C for 3 months. Example 8: Selection of protein, arginine, and polysorbate 80 concentrations A. Sample Preparation

[0214] Each candidate formulation (F1-F12) was prepared as follows. The drug substance (DS) processing intermediate was concentrated and equilibrated using the corresponding formulation buffer (Table 22) via centrifugation and filtration. After concentration and equilibration, PS80 dissolved in the formulation buffer was added to achieve the desired concentration, and the protein concentration was adjusted to the final concentration. Each candidate formulation was filled into vials with a filling volume of 0.5 mL. The candidate formulations (F1-F12) are shown in Table 22. Formulation F0 was evaluated as a control. Table 22 Formulations (F0) and Candidate Formulations F1-F12 a: F2 and F10 are the same formulations, prepared for different studies. b: When selecting the concentration of polysorbate 80, remove the drug solution from the drug product and fill it with 0.5 mL to match the filling volume with the container sealing system. B. Stability Scheme

[0215] The stability protocols are shown in Tables 23, 24, and 25. Once removed from storage, the vials were stored at 5°C until testing. Storage conditions were as follows: • Long-term conditions: Store at 5℃±3℃, upright. Acceleration conditions: Store at 25℃±2℃, 60%RH±5%RH, upright. • Stress conditions (freeze-thaw): Freeze at -30°C and thaw at room temperature, then place upright. • Stress conditions (stirring): 5℃, horizontal placement, vibration at 250 rpm on a reciprocating shaker, vibration amplitude of 50 mm. Table 23 Stability Schemes (Long-Term and Accelerated Conditions) N / A: Not applicable. Table 24 Stability Schemes (Stress Conditions: F0-F7) N / A: Not applicable. Table 25 Stability Schemes (Stress Conditions: F0, F8-F12) N / A: Not applicable.

[0216] Because the viscosity of solutions is known to increase exponentially in high-concentration protein solutions, the protein concentration of each candidate formulation (F1-F5) was adjusted to 200 ± 10 mg / mL using a formulation buffer containing 0.05 (w / v) % PS80 for analysis. Other formulations were not diluted prior to measurement. C. Results and Discussion (a) Selection of arginine concentration

[0217] To select the target arginine concentration in the formulation, physical properties were evaluated, and freeze-thaw, long-term, and accelerated stability studies were conducted on candidate formulations with different arginine concentrations (150 to 350 mmol / L, F1 to F5) compared to formulation F0 (see Table 22). The results of the physical properties and freeze-thaw studies are shown in Table 26. The results of the long-term and accelerated stability studies are shown in Tables 27 and 28. (b) Physical properties

[0218] As shown in Table 26, the osmolality values ​​increased with increasing arginine concentration. Compared to F4 and F5, F1, F2, and F3 showed lower osmolality values. Therefore, the arginine concentration was limited to 250 mmol / L or lower. The viscosity values ​​of F1–F5 were within a narrow range (7.3–8.1 cP) and were not correlated with arginine concentrations in the range of 150–350 mmol / L. The results for PS80, as well as the appearance, pH, and concentration of the protein, were almost entirely satisfactory. (c) Freeze-thaw stability study

[0219] As shown in Table 26, no significant changes were observed in any of the test items after three cycles of freeze-thaw testing. (d) Long-term stability study

[0220] As shown in Table 27, after three months of storage, no significant changes were observed in any of the test items except for size exclusion HPLC (SEC). In candidate formulations F1-F5, the amounts of aggregates and fragments measured by SEC all showed a slight increase. The rate of increase was similar to that of formulation F0. (e) Accelerated stability study

[0221] As shown in Table 28, no significant changes were observed in any of the tests except for SEC and ion-exclusion HPLC (IEX). Although the amount of aggregates measured by SEC increased in all candidates (F1-F5, BAN2401 200 mg / mL), and as expected, at a faster rate than in formulation F0, all candidates were considered feasible given the results under long-term conditions and the stability of F0 over its long shelf life. The higher arginine concentration resulted in slightly slower aggregate formation, consistent with the results described in Example 4. The amount of fragments measured by SEC also increased in all candidates, but at a rate similar to that in formulation F0. As for IEX, the amount of acidic peaks increased in all candidates, but at a rate similar to that in F0. (f) Arginine concentration

[0222] As shown in Table 26, F4 and F5 (arginine concentrations of 300 and 350 mmol / L) were deemed infeasible in terms of osmolality. Among the feasible candidates F1, F2, and F3 (arginine concentrations of 150, 200, and 250 mmol / L, respectively), F1 was closest to isotonicity. As shown in Table 28, formulations with higher arginine concentrations exhibited lower aggregate formation rates in accelerated stability studies, but this difference was not significant within the evaluated arginine concentration range. Considering isotonicity and aggregate formation rate, 200 mmol / L was selected as the target arginine concentration based on formulation F2. (g) Selection of protein concentration

[0223] To select the target protein concentration in the formulation, physical properties were evaluated, and freeze-thaw, long-term, and accelerated stability studies were conducted on candidate formulations and formulation F0 with different protein concentrations (200 to 300 mg / mL, F2, F6, F7). The results of the physical properties and freeze-thaw studies are shown in Table 29. The results of the long-term and accelerated stability studies are shown in Tables 27 and 28. (h) Physical properties

[0224] As shown in Table 29, F2 and F6 exhibit lower osmotic molar concentration values ​​compared to F7. The viscosity values ​​for F2, F6, and F7 are 7.8, 21, and 48 cP, respectively. High viscosities exceeding 20 cP can lead to difficulties in DS fabrication by increasing pump back pressure and reducing transmembrane flux during ultrafiltration and diafiltration steps. Furthermore, higher concentrations of solution can cause changes in packing weight by clogging the packing needle, further complicating the DP packing process. Since the desired viscosity is reported to be no more than 20 cP... 3 Therefore, F2 is feasible. The results for appearance, pH, and the concentrations of PS80 and protein almost met the targets. (i) Freeze-thaw study

[0225] As shown in Table 29, no significant changes were observed in any of the test items after three cycles of freeze-thaw testing. (j) Long-term stability study

[0226] As shown in Table 27, after three months of storage, no significant changes were observed except for size exclusion HPLC (SEC). In candidate formulations F2, F6, and F7, the amount of aggregates measured by SEC increased slightly, but the rate of increase was similar to that in F0. (k) Accelerated stability study

[0227] As shown in Table 28, no significant changes were observed except by SEC and ion-exclusion HPLC (IEX). At the three-month time point, the amount of aggregates measured by SEC increased at a similar rate (0.7%–0.8%) in all candidates (F2, F6, and F7). Therefore, all candidates were considered feasible. The amount of fragments measured by SEC also increased in all candidates, but at a rate similar to that in F0. As for IEX, the amount of acidic peaks obtained by IEX increased in all candidates, but at a rate similar to that in F0. (l) Protein concentration

[0228] Considering the results of osmolar concentration and viscosity shown in Table 29 and the overall stability results shown in Tables 27 and 28, a protein concentration of 200 mg / mL was selected. (m) Selection of PS80 concentration

[0229] To select the target PS80 concentration in the formulation, freeze-thaw and agitation studies were conducted on candidate formulations and formulation F0 with different PS80 concentrations [0 to 0.10 (w / v)%, F8-F12]. The results of the freeze-thaw and agitation studies are shown in Tables 30 and 31, respectively. (n) Freeze-thaw research

[0230] As shown in Table 30, no significant changes were observed in any of the test items after three cycles of freeze-thaw testing. In F8 (without PS80), the particle count obtained by MFI was relatively high. (o) Stirring Research

[0231] As shown in Table 31, formulations (F9 to F12) containing 0.02 (w / v)% to 0.10 (w / v)% PS80 were stable after stirring. After stirring, changes in appearance, protein concentration, and aggregate amount were observed by SEC for F8 (without PS80). The appearance changed from a clear pale yellow liquid to a milky white liquid. The protein concentration decreased slightly from 206 mg / mL to 195 mg / mL. The amount of aggregates increased from 1.1% to 3.8%. Therefore, F8 (without PS80) was considered unfeasible. (p)PS80 concentration

[0232] Based on the results shown in Tables 30 (Freeze-Thaw Study) and 31 (Agitation Study), formulations containing 0.02 (w / v)% to 0.10 (w / v)% PS80 were stable after three freeze-thaw cycles and agitation for up to three days. Therefore, a PS80 concentration of 0.05 (w / v)% was selected as the target. (q) Conclusion

[0233] In summary, the following formulations of BAN2401 were selected for further research. Table 32 Preparations used for subsequent studies Example 9: Stability study of 200 mg / mL BAN2401 formulations at different pH values

[0234] Stability studies were conducted to test the stability of the 200 mg / mL BAN2401 formulation at different pH values. Additionally, the effect of methionine addition on the stability of the pharmaceutical product (DP) was investigated. A. Sample Preparation

[0235] Each candidate formulation was prepared as follows (Table 33). The formulation was equilibrated with the appropriate formulation buffer via centrifugation filtration. For F20 preparation only, a formulation buffer with a pH of 3.5 was used until the pH of the filtrate reached 4.0, as protons are repelled by concentrated charged proteins near the semipermeable membrane; this is known as the Donnan effect. After concentration and equilibration, PS80 dissolved in the formulation buffer was added to reach the predetermined concentration, and then the protein concentration was adjusted to the final concentration. Each candidate formulation was filled into vials with a filling volume of 0.5 mL. Formulation F0 was evaluated as a control (see Table 33). Table 33 Composition of Active Pharmaceutical Ingredients and Candidate SC Formulations Arginine (Arginine), Met (Methionine) a: To adjust the pH, arginine and arginine hydrochloride are combined for F0; hydrochloride is added for F13 to F22. b: F21 is the same formulation as F15. B. Storage Plan

[0236] The storage plan is shown in Tables 34, 35, and 36. Once removed from storage, the vials should be stored at 5°C until the test begins. Storage conditions are as follows: • Long-term conditions: Store at 5℃±3℃, upright. Acceleration conditions: Store at 25℃±2℃, 60%RH±5%RH, upright. • Stress conditions (freeze-thaw): Freeze at -30°C and thaw at room temperature, then place upright. 1000 lx: 25℃±2℃, 60%RH±5%RH, with an exposure of 1000 lx, laid flat. Darkness: 25℃±2℃, 60%RH±5%RH, covered with aluminum foil without exposure, laid flat.

[0237] To confirm aggregate levels after three months or longer storage, size exclusion HPLC of samples stored for nine months was additionally evaluated for long-term stability studies, and size exclusion HPLC of samples stored for three months was evaluated for accelerated stability studies. The latter samples were tested after being stored in a refrigerator for six months. C. Results and Discussion

[0238] Physical properties were evaluated, and long-term and accelerated studies were conducted on candidate formulations, including those with pH variations (pH 4.0 to pH 6.0) and methionine addition. Freeze-thaw and photostable stability studies were also performed to assess the efficacy of methionine addition. D. pH changes

[0239] As shown in Tables 37, 38A and 38B, 39, and 40, no significant differences were observed among the candidate formulations (F13, F15, F17, and F20 through F22) for any test item, except for size exclusion HPLC (SEC). Formulation F20 (i.e., the lower pH formulation at pH 4.0) showed a lower amount of aggregates as measured by SEC at the initial time point. Furthermore, a slightly lower aggregate formation rate and a higher fragment formation rate were observed in the accelerated stability studies. Results for formulations with pH values ​​from 4.5 to 5.5 were confirmed to be similar. E. Addition of methionine

[0240] As shown in Tables 41A and 41B and Tables 42 through 46, no significant differences were observed between formulations containing or without methionine (F18 and F15). In extended long-term stability studies, accelerated stability studies, and photostability studies, the formulation containing methionine (F18) showed a slightly lower aggregate formation rate. F18 also showed slightly lower oxidation of the methionine residues (e.g., at position 259 in the heavy chain) after exposure to 1000 lux for up to seven days. Freeze-thaw studies showed no differences. The role of 10 mmol / L methionine as a stabilizer is limited. F. Conclusion

[0241] In BAN2401 formulations with a pH range of 4.5 to 5.5, the quality properties were similar. Formulation at pH 4.0 showed a lower aggregate formation rate, while fragment formation was faster during storage. Based on quality assessments, including long-term, accelerated, freeze-thaw, and light stability studies, we found no significant differences between formulations containing or without methionine at a concentration of 10 mmol / L. Although a slightly slower aggregate formation rate and less oxidative inhibition of amino acid residues were observed, the differences were not significant. In conclusion, formulations F15 and F21, composed of 200 mg / mL BAN2401, 25 mmol / L L-histidine / histidine hydrochloride, 200 mmol / L L-arginine, and 0.05 (w / v)% polysorbate 80 at pH 5.0 ± 0.5, were identified as promising drug substance candidates for further development. Example 10: Study on the effect of arginine concentration on the stability of BAN2401 formulation

[0242] The effect of arginine concentrations (0, 50, 100, and 200 mmol / L) on the stability of 200 mg / mL BAN2401 was evaluated. Samples at each arginine concentration level were stored under accelerated conditions (25 °C / 60% RH) and tested at 0, 1, and 2 months. A. Sample Preparation

[0243] Samples (F1-F4) were prepared by buffer exchange of concentrated BAN2401 active pharmaceutical ingredient processing intermediates using ultrafiltration. After aseptic filtration, each 0.4 mL sample was filled into a vial. Sample information is shown in Table 47. Table 47 Sample Information B. Result

[0244] Tables 48 to 51 show the results of the stability studies for samples F1 to F4. There were no differences in pH and protein concentration among the samples, and the arginine concentration levels were consistent with the target levels for the initial samples.

[0245] The aggregate percentage of samples containing arginine (F2 to F4) was lower than that of samples without arginine (F1). Table 48 shows the test results for sample F1 (for arginine 0 mmol / L). Table 49 shows the test results for sample F2 (for arginine 50 mmol / L). Table 50 shows the test results for sample F3 (for arginine at 100 mmol / L). Table 51 shows the test results for sample F4 (for arginine at 200 mmol / L). C. Discussion and Conclusion

[0246] The results of this study indicate that arginine concentrations of 50 to 200 mmol / L can prevent the increase of protein aggregates in BAN2401 200 mg / mL formulations. Variations in arginine concentration between batches may lead to variations in the percentage of aggregates between batches. Furthermore, in stability studies, arginine concentration may influence the trend of increasing aggregate percentages.

[0247] By comparing the fragmentation percentage in sample F1 with that in example 2 Figure 11 The percentage of fragmentation shown indicates that formulations containing histidine buffer appear to exhibit a reducing effect on BAN2401 fragmentation, with samples containing 200 mg / mL BAN2401, 50 mM sodium citrate, and 100 mM sodium chloride. sequence list Table 52. Amino acid sequences of the variable region of mAb Table 53. Amino acid sequence of the mAb constant region Table 54. Amino acid sequence of the mAb CDR Heavy chain (SEQ ID NO:11): evqlvesggglvqpggslrlscsasgftfssfgmhwvrqapgkglewvayissgsstiyygdtvkgrftisrdnaknslflqmsslraedtavyycareggyyygrsyytmdywgqgttvtvssastkgpsvfplapsskstsggtaalgclvkdyfpepvtvswnsgaltsgvhtfpavlqssglyslssvvtvpssslgtqtyicnvnhkpsntkvdkrvepkscdkthtcppcpapellggpsvflfppkpkdtlmisrtpevtcvvvdvshedpevkfnwyvdgvevhnaktkpreeqynstyrvvsvltvlhqdwlngkeykckvsnkalpapiektiskakgqprepqvytlppsreemtknqvsltclvkgfypsdiavewesngqpennykttppvldsdgsfflyskltvdksrwqqgnvfscsvmhealhnhytqkslslspgk Light chain (SEQ ID NO:12): dvvmtqsplslpvtpgapasiscrssqsivhsngntylewylqkpgqspklliykvsnrfsgvpdrfsgsgsgtdftlrisrveaedvgiyycfqgshvpptfgpgtkleikrtvaapsvfifppsdeqlksgtasvvcllnnfypreakvqwkvdnalqsgnsqesvteqdskdstyslsstltlskadyekhkvyacevthqglsspvtksfnrgec

Claims

1. An aqueous pharmaceutical preparation comprising: (a) Isolated anti-Aβ primary fibrillary antibodies that bind to human Aβ primary fibrils at concentrations ranging from 80 mg / mL to 240 mg / mL. (b) 100 mM to less than 300 mM of arginine, arginine hydrochloride, or a combination thereof. (c) Polysorbate 80 at 0.01% w / v to 0.1% w / v, and (d) 10 mM to 100 mM citrate buffer or 10 mM to 100 mM histidine buffer. The drug formulation has a pH range of 4.5 to 5.

5. The isolated anti-Aβ protofibrillary antibody is lecanemab.

2. The pharmaceutical formulation according to claim 1, wherein the isolated anti-Aβ primary fibrillary antibody is present at a concentration of 100 mg / mL to 200 mg / mL.

3. The pharmaceutical formulation according to claim 1, wherein the isolated anti-Aβ primary fibrillary antibody is present at a concentration of 100 mg / mL.

4. The pharmaceutical formulation according to claim 1, wherein the isolated anti-Aβ primary fibrillary antibody is present at a concentration of 200 mg / mL.

5. The pharmaceutical formulation according to any one of claims 1-4, wherein the pharmaceutical formulation further comprises methionine.

6. The pharmaceutical preparation according to any one of claims 1-4, wherein the pharmaceutical preparation comprises 125 mM to less than 300 mM of arginine, arginine hydrochloride, or a combination thereof.

7. The pharmaceutical preparation according to any one of claims 1-4, wherein the pharmaceutical preparation comprises 200 mM arginine hydrochloride.

8. The pharmaceutical preparation according to any one of claims 1-4, wherein the pharmaceutical preparation comprises 200 mM arginine hydrochloride and 25 mM histidine buffer.

9. The pharmaceutical formulation according to any one of claims 1-4, wherein the histidine buffer is a combination of histidine and histidine hydrochloride.

10. The pharmaceutical formulation according to any one of claims 1-4, wherein the histidine buffer is a combination of histidine and histidine hydrochloride monohydrate.

11. An aqueous pharmaceutical preparation comprising: (a) Isolated anti-Aβ primary fibrillary antibodies at concentrations ranging from 80 mg / mL to 240 mg / mL, (b) Arginine hydrochloride ranging from 140 mM to 260 mM (c) Polysorbate 80, from 0.02% w / v to 0.08% w / v, and (d) 15 mM to 35 mM histidine buffer The drug formulation has a pH range of 4.5 to 5.5, and The isolated anti-Aβ protofibrillary antibody is lecanemab.

12. An aqueous pharmaceutical preparation comprising: (a) Isolated anti-Aβ primary fibrillary antibodies at concentrations ranging from 80 mg / mL to 120 mg / mL, (b) 240 mM to less than 300 mM of arginine, arginine hydrochloride, or a combination thereof. (c) Polysorbate 80, from 0.02% w / v to 0.08% w / v, and (d) 30 mM to 50 mM citrate buffer The drug formulation has a pH range of 4.5 to 5.5, and The isolated anti-Aβ protofibrillary antibody is lecanemab.

13. The aqueous pharmaceutical preparation according to claim 1, wherein the aqueous pharmaceutical preparation comprises: (a) 100 mg / mL or 200 mg / mL of the isolated anti-Aβ primary fibrillary antibody, (b) 190 mM to 200 mM arginine hydrochloride, (c) Polysorbate 80 at 0.04% w / v to 0.06% w / v, and (d) 15 mM to 35 mM histidine buffer.

14. The aqueous pharmaceutical preparation according to claim 1, wherein the aqueous pharmaceutical preparation comprises: (a) 100 mg / mL of the isolated anti-Aβ primary fibrillary antibody, (b) 200 mM arginine hydrochloride, (c) 0.05% w / v polysorbate 80, and (d) 25 mM histidine buffer.

15. The aqueous pharmaceutical preparation according to claim 1, wherein the aqueous pharmaceutical preparation comprises: (a) 200 mg / mL of the isolated anti-Aβ primary fibrillary antibody, (b) 200 mM arginine hydrochloride, (c) 0.05% w / v polysorbate 80, and (d) 25 mM histidine buffer.

16. The aqueous pharmaceutical preparation according to any one of claims 13-15, wherein the histidine buffer is a combination of histidine and histidine hydrochloride monohydrate.

17. The aqueous pharmaceutical preparation according to any one of claims 13-15, wherein the pH of the pharmaceutical preparation is 5.0.

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