Stable antibody formulation

Abstract

To provide stable pharmaceutical formulations comprising a human antibody that specifically binds to Ebola Virus (EBOV).SOLUTION: A pharmaceutical formulation comprises: (a) a stabilizer; (b) a buffer comprising histidine; (c) an organic cosolvent comprising polysorbate; and (d) an antibody which binds specifically to Ebola virus (EBOV) and comprises (i) a first antibody comprising a specific amino acid sequence, (ii) a second antibody comprising a specific amino acid sequence, and (iii) a third antibody comprising a specific amino acid sequence, the formulation comprising (i), (ii), (iii), (i)+(ii), (i)+(iii), (ii)+(iii), or (i)+(ii)+(iii), and having a pH of 6.0±0.3.SELECTED DRAWING: None

Description

Technical Field

[0001] Government license rights This invention was made with government support under Contract Nos. HHS0100201500013C and HHS0100201700016C awarded by the U.S. Department of Health and Human Services. The government has certain rights in this invention.

[0002] Sequence Listing A public copy of the sequence listing was electronically submitted via EFS-Web as an ASCII-formatted sequence listing simultaneously with this specification. The file name of the copy is "10668WO01_Sequence_Listing_ST25.txt", the creation date is January 22, 2021, and the size is approximately 36 KB. The sequence listing contained in this ASCII-formatted written material is part of this specification and is hereby incorporated by reference in its entirety.

[0003] Field of the Invention The present disclosure relates to the field of therapeutic antibody formulations. More specifically, the present disclosure relates to the field of pharmaceutical formulations comprising one or more human antibodies that specifically bind to the Ebola virus (EBOV).

Background Art

[0004] Background Therapeutic polymers (e.g., antibodies) must be formulated in a way that not only makes them suitable for administration to patients but also maintains their stability during storage and subsequent use. For example, a therapeutic antibody in a liquid solution is prone to decomposition, aggregation, or undesirable chemical modifications if the solution is not properly formulated. The stability of an antibody in a liquid formulation depends not only on the type of excipients used in the formulation but also on the relative amounts and ratios of the excipients to each other. Furthermore, other considerations besides stability must be taken into account when preparing a liquid antibody formulation. Examples of such additional considerations include the viscosity of the solution and the concentration of the antibody that can be accommodated by a given formulation, as well as the visual quality or appeal of the formulation. Therefore, when formulating a therapeutic antibody, great care must be taken to obtain a formulation that maintains stability, contains an appropriate concentration of the antibody, has an appropriate viscosity, and has other properties that allow for easy administration of the formulation to patients. Antibodies against the Ebola virus (EBOV) are an example of a therapeutic-related polymer that requires proper formulation. Anti-EBOV antibodies are clinically useful for the prevention and / or treatment of Ebola virus infection. Examples of anti-EBOV antibodies are described, in particular, in U.S. Patent Nos. 10,501,526 (Patent Document 1), 10,081,670 (Patent Document 2), 9,771,414 (Patent Document 3), 6,630,144 (Patent Document 4), 6,875,433 (Patent Document 5), 7,335,356 (Patent Document 6), and 8,513,391 (Patent Document 7), as well as WO2016 / 123019 (Patent Document 8), EP1539238 (Patent Document 9), EP2350270 (Patent Document 10), and EP8513391 (Patent Document 11).

[0005] Although anti-EBOV antibodies are known, there remains a need for novel pharmaceutical formulations containing anti-EBOV antibodies that are sufficiently stable and suitable for administration to patients, including those in remote environments or in environments where the therapeutic agent cannot be refrigerated.

Prior Art Documents

Patent Documents

[0006] [Patent Document 1] U.S. Patent No. 10,501,526 [Patent Document 2] U.S. Patent No. 10,081,670 [Patent Document 3] U.S. Patent No. 9,771,414 [Patent Document 4] U.S. Patent No. 6,630,144 [Patent Document 5] U.S. Patent No. 6,875,433 [Patent Document 6] U.S. Patent No. 7,335,356 [Patent Document 7] U.S. Patent No. 8,513,391 [Patent Document 8] WO2016 / 123019 [Patent Document 9] EP1539238 [Patent Document 10] EP2350270 [Patent Document 11] EP8513391 [Summary of the Invention]

[0007] Summary The present disclosure meets the aforementioned needs by providing a stable pharmaceutical formulation comprising a human antibody that specifically binds to Ebola virus (EBOV).

[0008] In one aspect, there is provided a stable liquid pharmaceutical formulation comprising (a) a stabilizer containing a sugar, (b) a buffer containing histidine, (c) an organic co-solvent containing polysorbate, and (d) at least one antibody that specifically binds to Ebola virus (EBOV).

[0009] In various embodiments, at least one antibody that specifically binds to EBOV is provided at a concentration of about 5 ± 0.75 mg / mL to about 250 ± 37.5 mg / mL. In some embodiments, 250 mg / mL is the maximum protein concentration in the formulation. In some aspects, the 250 mg / mL of protein comprises up to three antibodies. In some aspects, the maximum protein concentration in a formulation comprising three antibodies ranges from about 5 ± 0.75 mg / mL to about 250 mg / mL ± 37.5 mg / mL.

[0010] The ratio of two or three antibodies present in the formulation can be adjusted according to the measured value of the result. In some aspects, the two antibodies are present in a 1:1 ratio. In some aspects, the antibodies are present in a 1:2 ratio. In some aspects, the two antibodies are present in a ratio of about 1 to 10:1. In some aspects, the three antibodies are present in a 1:1:1 ratio. In some aspects, the three antibodies are present in a 1:2:1 ratio. In some aspects, the three antibodies are present in a 2:1:1 ratio. In some aspects, the three antibodies are present in a 1:1:2 ratio. In some aspects, the antibodies are present in a ratio of about 1 to 10:1 to 10:1 to about 10.

[0011] In some embodiments, the dosage is about 3000 mg, about 2000 mg, about 1500 mg, 1000 mg, about 800 mg, about 750 mg, about 500 mg, about 250 mg, about 200 mg, about 150 mg, about 100 mg, about 75 mg, about 50 mg, or about 25 mg. In some aspects, the dosage comprises one anti-EBOV antibody. In some aspects, the dosage comprises two anti-EBOV antibodies. In some aspects, the dosage comprises three anti-EBOV antibodies. In one embodiment, the co-formulated antibody is delivered intravenously over about 2 hours.

[0012] In one embodiment, at least one antibody is provided at a concentration of 12.5 mg / mL ± 1.85 mg / mL, or about 12.5 mg / mL. In one embodiment, at least one antibody is provided at a concentration of 25 mg / mL ± 3.75 mg / mL, or about 25 mg / mL. In another embodiment, at least one antibody is provided at a concentration of 50 mg / mL ± 7.5 mg / mL, or about 50 mg / mL. In another embodiment, at least one antibody is provided at a concentration of 100 mg / mL ± 15 mg / mL, or about 100 mg / mL. In another embodiment, at least one antibody is provided at a concentration of 125 mg / mL ± 18.75 mg / mL, or about 125 mg / mL. In one embodiment, at least one antibody is provided at a concentration of 150 mg / mL ± 22.5 mg / mL, or about 150 mg / mL. In another embodiment, at least one antibody is provided at a concentration of 175 mg / mL ± 26.25 mg / mL, or about 175 mg / mL. In another embodiment, at least one antibody is provided at a concentration of 200 mg / mL ± 30 mg / mL, or about 200 mg / mL. In another embodiment, at least one antibody is provided at a concentration of 250 mg / mL ± 37.5 mg / mL, or about 250 mg / mL.

[0013] In some embodiments, each antibody is administered at 50 mg / kg body weight. In one embodiment, three antibodies are co-formulated such that the final formulation is administered at 50 mg / kg body weight for each antibody. Thus, the final dose administered to the patient is 150 mg / kg body weight, and the three antibodies in the formulation are in a 1:1:1 ratio. In one embodiment, the co-formulated antibodies are delivered intravenously over about 2 hours.

[0014] In certain embodiments, it comprises any one or more of the anti-EBOV antibodies disclosed in US Patent Application Publication 2016 / 0215040, which is incorporated herein by reference in its entirety. In certain embodiments, the anti-EBOV antibody comprises (a) a heavy chain variable region (HCVR) comprising heavy chain complementarity determining regions 1, 2, and 3 (HCDR1-HCDR2-HCDR3) each comprising the sequences of SEQ ID NO: 4, SEQ ID NO: 6, and SEQ ID NO: 8, respectively, and (b) a light chain variable region (LCVR) comprising light chain complementarity determining regions 1, 2, and 3 (LCDR1-LCDR2-LCDR3) each comprising the sequences of SEQ ID NO: 12, SEQ ID NO: 14, and SEQ ID NO: 16, respectively. In one embodiment, the antibody comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 2 and an LCVR comprising the amino acid sequence of SEQ ID NO: 10. In some aspects, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 17 and a light chain comprising the amino acid sequence of SEQ ID NO: 18. In one embodiment, the antibody comprises an HCVR having at least 90% sequence identity to SEQ ID NO: 2. In one embodiment, the antibody comprises an LCVR having at least 90% sequence identity to SEQ ID NO: 10. In one embodiment, the antibody comprises an HCVR having at least 95% sequence identity to SEQ ID NO: 2 and an LCVR having at least 95% sequence identity to SEQ ID NO: 10.

[0015] In certain embodiments, the anti-EBOV antibody comprises (a) a heavy chain variable region (HCVR) comprising heavy chain complementarity determining regions 1, 2, and 3 (HCDR1-HCDR2-HCDR3) each comprising the sequences of SEQ ID NO: 22, SEQ ID NO: 24, and SEQ ID NO: 26, respectively, and (b) a light chain variable region (LCVR) comprising light chain complementarity determining regions 1, 2, and 3 (LCDR1-LCDR2-LCDR3) each comprising the sequences of SEQ ID NO: 30, SEQ ID NO: 32, and SEQ ID NO: 34, respectively. In one embodiment, the antibody comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 20 and an LCVR comprising the amino acid sequence of SEQ ID NO: 28. In some aspects, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 35 and a light chain comprising the amino acid sequence of SEQ ID NO: 36. In one embodiment, the antibody comprises an HCVR having 90% sequence identity to SEQ ID NO: 20. In one embodiment, the antibody comprises an LCVR having 90% sequence identity to SEQ ID NO: 28. In one embodiment, the antibody comprises an HCVR having 95% sequence identity to SEQ ID NO: 20 and an LCVR having 95% sequence identity to SEQ ID NO: 28.

[0016] In certain embodiments, the anti-EBOV antibody comprises a heavy chain variable region (HCVR) comprising heavy chain complementarity determining regions 1, 2, and 3 (HCDR1-HCDR2-HCDR3) comprising the sequences of SEQ ID NO: 40, SEQ ID NO: 42, and SEQ ID NO: 44, respectively, each determining region, and (b) a light chain variable region (LCVR) comprising light chain complementarity determining regions 1, 2, and 3 (LCDR1-LCDR2-LCDR3) comprising the sequences of SEQ ID NO: 48, SEQ ID NO: 50, and SEQ ID NO: 52, respectively. In one embodiment, the antibody comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 38 and an LCVR comprising the amino acid sequence of SEQ ID NO: 46. In some aspects, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 53 and a light chain comprising the amino acid sequence of SEQ ID NO: 54. In one embodiment, the antibody comprises an HCVR having 90% sequence identity to SEQ ID NO: 38. In one embodiment, the antibody comprises an LCVR having 90% sequence identity to SEQ ID NO: 46. In one embodiment, the antibody comprises an HCVR having 95% sequence identity to SEQ ID NO: 38 and an LCVR having 95% sequence identity to SEQ ID NO: 46.

[0017] In one embodiment, the pH of the liquid formulation is pH 6.0 ± 0.5, pH 6.0 ± 0.4, pH 6.0 ± 0.3, pH 6.0 ± 0.2, pH 6.0 ± 0.1, pH 6.0 ± 0.05, pH 6.0 ± 0.01, or pH 6.0. In one embodiment, the pH of the liquid formulation is about pH 6.0 ± 0.3.

[0018] In one embodiment, the buffer contains histidine. In certain embodiments, the histidine buffer has a concentration of 5 mM ± 1 mM to 50 mM ± 10 mM, or 5 mM ± 1 mM to 25 mM ± 5 mM. In one embodiment, the histidine buffer has a concentration of 10 mM ± 2 mM or about 10 mM. In one embodiment, the histidine buffer has a concentration of 20 mM ± 4 mM or about 20 mM. In one embodiment, the histidine buffer has a concentration of 40 nM ± 8 mM or about 40 nM. In certain embodiments, the histidine buffer contains L-histidine and L-histidine monohydrochloride monohydrate. In one embodiment, L-histidine has a concentration of 2 mM ± 0.4 mM to 25 mM ± 5 mM, preferably 4 mM ± 0.8 mM to 20 mM ± 4 mM. In one embodiment, L-histidine monohydrochloride monohydrate has a concentration of 2 mM ± 0.4 mM to 25 mM ± 5 mM, preferably 4 mM ± 0.8 mM to 20 mM ± 4 mM. In one embodiment, the buffer contains L-histidine at a concentration of 4.8 mM ± 0.96 mM and L-histidine monohydrochloride monohydrate at a concentration of 5.2 mM ± 1.04 mM. In one embodiment, the buffer contains histidine at a concentration of 10 mM ± 2 mM, in which case the histidine contains L-histidine at a concentration of 4.8 mM ± 0.96 mM and L-histidine monohydrochloride monohydrate at a concentration of 5.2 mM ± 1.04 mM.

[0019] In certain embodiments, the organic co-solvent is a non-ionic polymer containing a polyoxyethylene moiety. In one embodiment, the organic co-solvent is a surfactant. In some embodiments, the organic co-solvent is any one or more of polysorbate, poloxamer 188, and polyethylene glycol 3350. In one embodiment, the organic co-solvent is polysorbate 80. In one embodiment, the organic co-solvent is polysorbate 20.

[0020] In one embodiment, the organic co-solvent is at a concentration of about 0.01% ± 0.005% to about 1% ± 0.5% "weight per volume" or "w / v", for example, 0.1 g / ml = 10% and 0.01 g / ml = 1%. In certain embodiments, the organic co-solvent is polysorbate at a concentration of 0.05% ± 0.025% to 0.5% ± 0.25% (w / v). In one embodiment, the organic co-solvent is polysorbate 80 at a concentration of 0.2% ± 0.1% w / v, or about 0.2%. In another embodiment, the organic co-solvent is polysorbate 80 at a concentration of 0.1% ± 0.05% w / v or about 0.1% w / v. In one embodiment, the organic co-solvent is polysorbate 20 at a concentration of 0.2% ± 0.1% w / v, or about 0.2%. In another embodiment, the organic co-solvent is polysorbate 20 at a concentration of 0.1% ± 0.05% w / v, or about 0.1% w / v.

[0021] In certain embodiments, the stabilizer is sugar. In one embodiment, the sugar is sucrose. In various embodiments, the stabilizer is at a concentration of 1% ± 0.2% w / v to 20% ± 4% w / v, 5% ± 1% w / v to 15% ± 3% w / v, or 1% ± 0.2% to 10% ± 2% w / v. In one embodiment, the stabilizer is sucrose at a concentration of 5% ± 1% w / v or about 5% w / v. In another embodiment, the stabilizer is sucrose at a concentration of 9% ± 1.8% w / v or about 9% w / v. In another embodiment, the stabilizer is sucrose at a concentration of 10% ± 2% w / v or about 10% w / v.

[0022] In certain embodiments, the formulation does not require a viscosity modifier. That is, the formulation does not contain a viscosity modifier. In certain embodiments, the formulation contains a viscosity modifier. In one embodiment, the formulation contains a viscosity modifier, and the viscosity modifier is an amino acid or a salt. In one embodiment, the viscosity modifier is L-proline. In certain embodiments, the viscosity modifier has a concentration of 1% ± 0.2% to 5% ± 1% w / v. In one embodiment, the viscosity modifier is proline at a concentration of 1.5% ± 0.3% or about 1.5%. In one embodiment, the viscosity modifier is proline at a concentration of 3% ± 0.6% or about 3%.

[0023] In certain embodiments, the viscosity of the liquid pharmaceutical formulation at 25°C is about 15 cPoise ± 10% or less. In certain embodiments, the viscosity at 25°C is 1.0 cPoise ± 10% to 20 cPoise ± 10%. In certain embodiments, the viscosity of the liquid pharmaceutical formulation is ≦ 20 cPoise. In certain embodiments, the viscosity of the liquid pharmaceutical formulation is ≦ 15 cPoise. In certain embodiments, the viscosity of the liquid pharmaceutical formulation is ≦ 10 cPoise. In certain embodiments, the viscosity of the liquid pharmaceutical formulation is ≦ 5 cPoise. In certain embodiments, the viscosity of the liquid pharmaceutical formulation is ≦ 2.5 cPoise. In certain embodiments, the viscosity at 25°C is about 2 cPoise ± 10%, 5 cPoise ± 10%, 6.0 cPoise ± 10%, 7.0 cPoise ± 10%, 7.1 cPoise ± 10%, 7.2 cPoise ± 10%, 7.9 cPoise ± 10%, 8.3 cPoise ± 10%, 9.0 cPoise ± 10%, 9.6 cPoise ± 10%, 10.0 cPoise ± 10%, 10.6 cPoise ± 10%, 11.4 cPoise ± 10%, 11.6 cPoise ± 10%, 11.8 cPoise ± 10%, 12.0 cPoise ± 10%, 13.0 cPoise ± 10%, 14.0 cPoise ± 10%, 15.0 cPoise ± 10%, or 16 cPoise ± 10%. In certain embodiments, the viscosity at 25°C is about 2.2 cPoise.

[0024] In one embodiment, a stable liquid pharmaceutical formulation is provided that contains, at pH 6.0 ± 0.3, (a) 5% ± 1% to 15% ± 3% w / v sucrose, (b) 5 mM ± 1 mM to 20 mM ± 4 mM histidine buffer, (c) 0.01% ± 0.005% to 0.5% ± 0.25% w / v polysorbate, such as about 0.05% ± 0.025% to about 0.2% ± 0.1% w / v polysorbate, and (d) 50 mg / mL ± 7.5 mg / mL total anti-EBOV antibody. In another embodiment, a stable liquid pharmaceutical formulation is provided that contains, at pH 6.0 ± 0.3, (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 50 mg / mL ± 7.5 mg / mL total antibody. In another embodiment, a stable liquid pharmaceutical formulation is provided that contains, at pH 6.0 ± 0.3, (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 50 mg / mL ± 5 g / mL total antibody.

[0025] In one embodiment, a stable liquid pharmaceutical formulation is provided that contains, at pH 6.0 ± 0.3, (a) 5% ± 1% to 15% ± 3% w / v sucrose, (b) 5 mM ± 1 mM to 20 mM ± 4 mM histidine buffer, (c) 0.01% ± 0.005% to 0.5% ± 0.25% w / v polysorbate, and (d) 100 mg / mL ± 15 mg / mL total antibody. In another embodiment, a stable liquid pharmaceutical formulation is provided that contains, at pH 6.0 ± 0.3, (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 100 mg / mL ± 15 mg / mL total antibody. In another embodiment, a stable liquid pharmaceutical formulation is provided that contains, at pH 6.0 ± 0.3, (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 100 mg / mL ± 10 mg / mL total antibody.

[0026] The anti-EBOV antibody comprises at least one antibody containing three heavy chain complementarity determining regions (CDRs) (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in a heavy chain variable region / light chain variable region (HCVR / LCVR) amino acid sequence pair selected from the group consisting of (i) SEQ ID NO: 2 / 10, (ii) SEQ ID NO: 20 / 28, and (iii) SEQ ID NO: 38 / 46. In some embodiments, the formulation comprises (i), (ii), (iii), (i)+(ii), (i)+(iii), (ii)+(iii), or (i)+(ii)+(iii), and the total anti-EBOV antibody present in the formulation is 50 mg / mL ± 7.5 mg / mL. In some embodiments, the formulation comprises (i), (ii), (iii), (i)+(ii), (i)+(iii), (ii)+(iii), or (i)+(ii)+(iii), and the total anti-EBOV antibody present in the formulation is 100 mg / mL ± 15 mg / mL.

[0027] In some embodiments, at least one anti-EBOV antibody comprises a heavy chain variable region (HCVR) and a light chain variable region (LCVR) such that the HCVR / LCVR combination comprises heavy chain complementarity determining regions and light chain complementarity determining regions (HCDR1-HCDR2-HCDR3 / LCDR1-LCDR2-LCDR3), and the combination comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 4-6-8 / 12-14-16, SEQ ID NO: 22-24-26 / 30-32-34, and SEQ ID NO: 40-42-44 / 48-50-52, respectively. In one embodiment, the anti-EBOV antibody comprises an amino acid sequence pair of a heavy chain variable region (HCVR) / light chain variable region (LCVR) selected from the group consisting of SEQ ID NO: 2 / 10, 20 / 28, and 38 / 46. In certain embodiments, the anti-EBOV antibody comprises an Fc region selected from the group consisting of human IgG1, IgG2, IgG3, and IgG4 isotypes. In one embodiment, the antibody comprises the human IgG1 isotype. In one embodiment, the antibody comprises a heavy chain comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 17, 35, and 53, and a light chain comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 18, 36, and 54. In one embodiment, the antibody has a molecular weight of 145 kDa ± 15 kDa, for example, 144,804 Da, 145,905 Da, or 143,689 Da.

[0028] In certain embodiments, a stable liquid pharmaceutical formulation is provided that comprises, at pH 6.0 ± 0.3, (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 50 mg / mL ± 7.5 mg / mL total antibody that specifically binds to EBOV, where the antibody comprises (i) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 2 / 10. In one embodiment, the anti-EBOV antibody comprises a HCVR comprising the amino acid sequence of SEQ ID NO: 2 and a LCVR comprising the amino acid sequence of SEQ ID NO: 10. In some aspects, the anti-EBOV antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 17 and a light chain comprising the amino acid sequence of SEQ ID NO: 18.

[0029] In certain embodiments, a stable liquid pharmaceutical formulation is provided that comprises, at pH 6.0 ± 0.3, (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 50 mg / mL ± 7.5 mg / mL total antibody that specifically binds to EBOV, where the antibody comprises (ii) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 20 / 28. In one embodiment, the anti-EBOV antibody comprises a HCVR comprising the amino acid sequence of SEQ ID NO: 20 and a LCVR comprising the amino acid sequence of SEQ ID NO: 28. In some aspects, the anti-EBOV antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 35 and a light chain comprising the amino acid sequence of SEQ ID NO: 36.

[0030] In certain embodiments, a stable liquid pharmaceutical formulation is provided that comprises, at pH 6.0 ± 0.3, (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 50 mg / mL ± 7.5 mg / mL total antibody that specifically binds to EBOV, wherein the antibody comprises (iii) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 38 / 46. In one embodiment, the anti-EBOV antibody comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 38 and an LCVR comprising the amino acid sequence of SEQ ID NO: 46. In some aspects, the anti-EBOV antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 53 and a light chain comprising the amino acid sequence of SEQ ID NO: 54.

[0031] In certain embodiments, a stable liquid pharmaceutical formulation is provided that comprises, at pH 6.0 ± 0.3, (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 50 mg / mL ± 7.5 mg / mL total antibody that specifically binds to EBOV, wherein the first antibody comprises (i) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 2 / 10, and the second antibody comprises (ii) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 20 / 28. In one embodiment, the first anti-EBOV antibody comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 2 and an LCVR comprising the amino acid sequence of SEQ ID NO: 10, and the second anti-EBOV antibody comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 20 and an LCVR comprising the amino acid sequence of SEQ ID NO: 28.

[0032] In certain embodiments, a stable liquid pharmaceutical formulation is provided that comprises, at pH 6.0 ± 0.3, (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) a total antibody of 50 mg / mL ± 7.5 mg / mL that specifically binds to EBOV, where the first antibody comprises (i) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 2 / 10, and the second antibody comprises (iii) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 38 / 46. In one embodiment, the first anti-EBOV antibody comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 2 and an LCVR comprising the amino acid sequence of SEQ ID NO: 10, and the second anti-EBOV antibody comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 38 and an LCVR comprising the amino acid sequence of SEQ ID NO: 46.

[0033] In certain embodiments, a stable liquid pharmaceutical formulation is provided that comprises, at pH 6.0 ± 0.3, (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 50 mg / mL ± 7.5 mg / mL total antibody that specifically binds to EBOV, wherein the first antibody comprises (ii) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 20 / 28, and the second antibody comprises (iii) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 38 / 46. In one embodiment, the first anti-EBOV antibody comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 20 and an LCVR comprising the amino acid sequence of SEQ ID NO: 28, and the second anti-EBOV antibody comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 38 and an LCVR comprising the amino acid sequence of SEQ ID NO: 46.

[0034] In certain embodiments, a stable liquid pharmaceutical formulation is provided that contains, at pH 6.0 ± 0.3, (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 50 mg / mL ± 7.5 mg / mL total antibody that specifically binds to EBOV, where the first antibody comprises (i) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 2 / 10, the second antibody comprises (ii) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 20 / 28, and the third antibody comprises (iii) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 38 / 46. In one embodiment, the first anti-EBOV antibody comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 2 and an LCVR comprising the amino acid sequence of SEQ ID NO: 10, the second anti-EBOV antibody comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 20 and an LCVR comprising the amino acid sequence of SEQ ID NO: 28, and the third anti-EBOV antibody comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 38 and an LCVR comprising the amino acid sequence of SEQ ID NO: 46.

[0035] In certain embodiments, a stable liquid pharmaceutical formulation is provided that comprises, at pH 6.0 ± 0.3, (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 100 mg / mL ± 15 mg / mL total antibody that specifically binds to EBOV, wherein the antibody comprises (i) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 2 / 10. In one embodiment, the anti-EBOV antibody comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 2 and an LCVR comprising the amino acid sequence of SEQ ID NO: 10. In some aspects, the anti-EBOV antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 17 and a light chain comprising the amino acid sequence of SEQ ID NO: 18.

[0036] In certain embodiments, a stable liquid pharmaceutical formulation is provided that comprises, at pH 6.0 ± 0.3, (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 100 mg / mL ± 15 mg / mL total antibody that specifically binds to EBOV, wherein the antibody comprises (ii) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 20 / 28. In one embodiment, the anti-EBOV antibody comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 20 and an LCVR comprising the amino acid sequence of SEQ ID NO: 28. In some aspects, the anti-EBOV antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 35 and a light chain comprising the amino acid sequence of SEQ ID NO: 36.

[0037] In certain embodiments, a stable liquid pharmaceutical formulation is provided that comprises, at pH 6.0 ± 0.3, (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 100 mg / mL ± 15 mg / mL total antibody that specifically binds to EBOV, wherein the antibody comprises (iii) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 38 / 46. In one embodiment, the anti-EBOV antibody comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 38 and an LCVR comprising the amino acid sequence of SEQ ID NO: 46. In some aspects, the anti-EBOV antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 53 and a light chain comprising the amino acid sequence of SEQ ID NO: 54.

[0038] In certain embodiments, a stable liquid pharmaceutical formulation is provided that comprises, at pH 6.0 ± 0.3, (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 100 mg / mL ± 15 mg / mL total antibody that specifically binds to EBOV, wherein the first antibody comprises (i) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 2 / 10, and the second antibody comprises (ii) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 20 / 28. In one embodiment, the first anti-EBOV antibody comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 2 and an LCVR comprising the amino acid sequence of SEQ ID NO: 10, and the second anti-EBOV antibody comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 20 and an LCVR comprising the amino acid sequence of SEQ ID NO: 28.

[0039] In certain embodiments, a stable liquid pharmaceutical formulation is provided that comprises, at pH 6.0 ± 0.3, (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 100 mg / mL ± 15 mg / mL total antibody that specifically binds to EBOV, wherein the first antibody comprises (i) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 2 / 10, and the second antibody comprises (iii) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 38 / 46. In one embodiment, the first anti-EBOV antibody comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 2 and an LCVR comprising the amino acid sequence of SEQ ID NO: 10, and the second anti-EBOV antibody comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 38 and an LCVR comprising the amino acid sequence of SEQ ID NO: 46.

[0040] In certain embodiments, a stable liquid pharmaceutical formulation is provided that comprises, at pH 6.0 ± 0.3, (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 100 mg / mL ± 15 mg / mL total antibody that specifically binds to EBOV, wherein the first antibody comprises (ii) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 20 / 28, and the second antibody comprises (iii) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 38 / 46. In one embodiment, the first anti-EBOV antibody comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 20 and an LCVR comprising the amino acid sequence of SEQ ID NO: 28, and the second anti-EBOV antibody comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 38 and an LCVR comprising the amino acid sequence of SEQ ID NO: 46.

[0041] In certain embodiments, a stable liquid pharmaceutical formulation is provided that comprises, at pH 6.0 ± 0.3, (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 100 mg / mL ± 15 mg / mL total antibody that specifically binds to EBOV, wherein the first antibody comprises (i) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 2 / 10, the second antibody comprises (ii) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 20 / 28, and the third antibody comprises (iii) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 38 / 46. In one embodiment, the first anti-EBOV antibody comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 2 and an LCVR comprising the amino acid sequence of SEQ ID NO: 10, the second anti-EBOV antibody comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 20 and an LCVR comprising the amino acid sequence of SEQ ID NO: 28, and the third anti-EBOV antibody comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 38 and an LCVR comprising the amino acid sequence of SEQ ID NO: 46.

[0042] In certain embodiments, a formulation of any of the foregoing aspects has a property selected from the group consisting of: (i) as described herein, the formulation is stable to long-term storage at 60°C, 55°C, 50°C, 45°C, 40°C, 35°C, 30°C, 25°C, 5°C, -20°C, -30°C, and -80°C; (ii) as described herein, the formulation is stable to agitation stress; (iii) as described herein, the formulation is stable to heat stress; (iv) the formulation has a low viscosity (less than 20 cPoise viscosity, preferably less than 15 cPoise viscosity); (v) as described herein, the formulation is stable even if there is up to a ±50% variation in the formulation excipient concentration; (vi) the formulation is iso-osmolar to physiological conditions; (vii) the formulation is stable and compatible with intravenous delivery devices and procedures; and (viii) the formulation is stable to long-term storage in glass vials or prefilled syringes.

[0043] In certain embodiments, a formulation of any of the foregoing aspects has a property selected from the group consisting of: (i) the formulation has a viscosity of less than 10 cP; (ii) the formulation has a viscosity of less than 5 cP; (iii) the formulation has a viscosity of less than about 2.5; (iv) at least 90% of the antibody is the native species after 28 days at 45°C; (v) at least 18% of the antibody is the major charge variant of the antibody after 28 days at 45°C; (vi) at least 96% of the antibody is the native species after 3 months at 25°C; (vii) at least 30% of the antibody is the major charge variant of the antibody after 3 months at 25°C; (viii) at least 96% of the antibody is the native species after 36 months at 5°C; (ix) at least 34% of the antibody is the major charge variant of the antibody after 36 months at 5°C; (x) at least 97% of the antibody is the native species after 120 minutes of agitation; (xi) at least 35% of the antibody is the major charge variant of the antibody after 120 minutes of agitation; (xii) at least 97% of the antibody is the native species after 8 freeze-thaw cycles; and / or (xiii) at least 35% of the antibody is the major charge variant of the antibody after 8 freeze-thaw cycles.

[0044] In certain embodiments, the antibody in the formulation of any of the foregoing aspects has a property selected from the group consisting of: the antibody retains at least about 90% of its ADCC activity after storage at -80°C for 12 months as compared to the activity of the same antibody prior to storage, (ii) the antibody retains at least about 80% or at least about 90% of its ADCC activity after storage at -30°C for 12 months as compared to the activity of the same antibody prior to storage, (iii) the antibody retains at least about 90% or at least about 95% of its ADCC activity after storage at -20°C for 3 months as compared to the activity of the same antibody prior to storage, (iv) the antibody retains at least about 90% or at least about 95% of its ADCC activity after storage at 5°C for 56 days as compared to the activity of the same antibody prior to storage, (v) the antibody retains at least about 90% or at least about 95% of its ADCC activity after storage at 25°C / 60% relative humidity (RH) for 28 days as compared to the activity of the same antibody prior to storage, (vi) the antibody retains at least about 90% or at least about 95% of its ADCC activity after storage at 40°C / 75% RH for 28 days as compared to the activity of the same antibody prior to storage, (vii) the antibody retains at least about 90% or at least about 95% of its ADCC activity after 120 minutes of agitation or after 8 freeze / thaw cycles, respectively, as compared to the activity of the same antibody prior to agitation or prior to freeze / thaw.

[0045] In certain embodiments, the antibody in the formulation of any of the foregoing aspects has a property selected from the group consisting of: the antibody retains at least about 90% or at least about 95% of its pseudovirus neutralizing activity after storage at -80°C for 12 months compared to the activity of the same antibody before storage, (ii) the antibody retains at least about 90% or at least about 95% of its pseudovirus neutralizing activity after storage at -30°C for 12 months compared to the activity of the same antibody before storage, (iii) the antibody retains at least about 90% or at least about 95% of its pseudovirus neutralizing activity after storage at -20°C for 3 months compared to the activity of the same antibody before storage, (iv) the antibody retains at least 80% or at least about 85% or at least about 90% or at least about 95% of its pseudovirus neutralizing activity after storage at 5°C for 56 days compared to the activity of the same antibody before storage, (v) the antibody retains at least about 80% or at least about 85% or at least about 90% or at least about 95% of its pseudovirus neutralizing activity after storage at 25°C / 60% relative humidity (RH) for 28 days compared to the activity of the same antibody before storage, (vi) the antibody retains at least about 80% or at least about 85% or at least about 90% or at least about 95% of its pseudovirus neutralizing activity after storage at 40°C / 75% RH for 28 days compared to the activity of the same antibody before storage, (vii) the antibody retains at least about 90% or at least about 95% of its pseudovirus neutralizing activity after 120 minutes of agitation or after 8 freeze / thaw cycles, respectively, compared to the activity of the same antibody before agitation or before freeze / thaw.

[0046] In certain embodiments of the aspect, there is provided a stable liquid formulation comprising: (a) at least one antibody that specifically binds to EBOV, said antibody comprising a heavy chain variable region / light chain variable region (HCVR / LCVR) amino acid sequence pair selected from the group consisting of (i) SEQ ID NO: 2 / 10, (ii) SEQ ID NO: 20 / 28, and (iii) SEQ ID NO: 38 / 46, and having a total antibody content of 50 mg / mL ± 7.5 mg / mL; (b) 10 mM ± 2 mM histidine buffer, pH 6.0 ± 0.3; (c) 0.1% ± 0.05% w / v polysorbate 80; and (d) 10% ± 2% w / v sucrose. In one embodiment according to the aspect, at least 96% of the antibody is an undenatured species after 36 months at 5°C. In one embodiment according to the aspect, at least 97% of the antibody is an undenatured species after 120 minutes of agitation. In one embodiment according to the aspect, at least 97% of the antibody is an undenatured species after 8 freeze-thaw cycles. In some aspects, the pharmaceutical formulation consists of (a) 50 mg / mL ± 7.5 mg / mL total antibody, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate 80, and (d) 10% ± 2% w / v sucrose in water at pH 6.0 ± 0.3.

[0047] In certain embodiments of the aspect, there is provided a stable liquid formulation comprising: (a) at least two antibodies that specifically bind to EBOV, wherein the antibodies comprise a heavy chain variable region / light chain variable region (HCVR / LCVR) amino acid sequence pair selected from the group consisting of (i) SEQ ID NO: 2 / 10, (ii) SEQ ID NO: 20 / 28, and (iii) SEQ ID NO: 38 / 46, and the total antibody comprises 50 mg / mL ± 7.5 mg / mL; (b) 10 mM ± 2 mM histidine buffer, pH 6.0 ± 0.3; (c) 0.1% ± 0.05% w / v polysorbate 80; and (d) 10% ± 2% w / v sucrose. In one embodiment according to the aspect, at least 96% of the antibody is the native species after 12 months at 5°C. In one embodiment according to the aspect, at least 97% of the antibody is the native species after 120 minutes of agitation. In one embodiment according to the aspect, at least 97% of the antibody is the native species after 8 freeze-thaw cycles. In some aspects, the pharmaceutical formulation consists of (a) 50 mg / mL ± 7.5 mg / mL total antibody, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate 80, and (d) 10% ± 2% w / v sucrose in water at pH 6.0 ± 0.3.

[0048] In certain embodiments of this aspect, there is provided a stable liquid formulation comprising: (a) a combination of three antibodies that specifically bind to EBOV, wherein the three antibodies comprise (i) the heavy chain variable region / light chain variable region (HCVR / LCVR) amino acid sequence pairs of SEQ ID NO: 2 / 10, (ii) SEQ ID NO: 20 / 28, and (iii) SEQ ID NO: 38 / 46, and the total antibody content is 50 mg / mL ± 7.5 mg / mL; (b) 10 mM ± 2 mM histidine buffer, pH 6.0 ± 0.3; (c) 0.1% ± 0.05% w / v polysorbate 80; and (d) 10% ± 2% w / v sucrose. In one embodiment according to this aspect, at least 96% of the antibodies are in the native form after 12 months at 5°C. In one embodiment according to this aspect, at least 97% of the antibodies are in the native form after 120 minutes of agitation. In one embodiment according to this aspect, at least 97% of the antibodies are in the native form after 8 freeze-thaw cycles. In some aspects, the pharmaceutical formulation consists of (a) 50 mg / mL ± 7.5 mg / mL total antibody, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate 80, and (d) 10% ± 2% w / v sucrose in water at pH 6.0 ± 0.3.

[0049] In certain embodiments of this aspect, there is provided a stable liquid formulation comprising: (a) at least one antibody that specifically binds to EBOV, wherein the antibody comprises a heavy chain variable region / light chain variable region (HCVR / LCVR) amino acid sequence pair selected from the group consisting of (i) SEQ ID NO: 2 / 10, (ii) SEQ ID NO: 20 / 28, and (iii) SEQ ID NO: 38 / 46, and the total antibody comprises 100 mg / mL ± 15 mg / mL; (b) 10 mM ± 2 mM histidine buffer, pH 6.0 ± 0.3; (c) 0.1% ± 0.05% w / v polysorbate 80; and (d) 10% ± 2% w / v sucrose. In one embodiment according to this aspect, at least 96% of the antibody is the native species after 12 months at 5°C. In one embodiment according to this aspect, at least 97% of the antibody is the native species after 120 minutes of agitation. In one embodiment according to this aspect, at least 97% of the antibody is the native species after 8 freeze-thaw cycles. In some aspects, the pharmaceutical formulation consists of (a) 100 mg / mL ± 15 mg / mL total antibody, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate 80, and (d) 10% ± 2% w / v sucrose in water at pH 6.0 ± 0.3.

[0050] In certain embodiments of this aspect, there is provided a stable liquid formulation comprising: (a) at least two antibodies that specifically bind to EBOV, said antibodies comprising a heavy chain variable region / light chain variable region (HCVR / LCVR) amino acid sequence pair selected from the group consisting of (i) SEQ ID NO: 2 / 10, (ii) SEQ ID NO: 20 / 28, and (iii) SEQ ID NO: 38 / 46, and having a total antibody content of 100 mg / mL ± 15 mg / mL; (b) 10 mM ± 2 mM histidine buffer, pH 6.0 ± 0.3; (c) 0.1% ± 0.05% w / v polysorbate 80; and (d) 10% ± 2% w / v sucrose. In one embodiment according to this aspect, at least 96% of the antibody is in the native species after 12 months at 5°C. In one embodiment according to this aspect, at least 97% of the antibody is in the native species after 120 minutes of agitation. In one embodiment according to this aspect, at least 97% of the antibody is in the native species after 8 freeze-thaw cycles. In some aspects, the pharmaceutical formulation consists of (a) 100 mg / mL ± 15 mg / mL total antibody, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate 80, and (d) 10% ± 2% w / v sucrose in water at pH 6.0 ± 0.3.

[0051] In certain embodiments of this aspect, there is provided a stable liquid formulation comprising: (a) a combination of three antibodies that specifically bind to EBOV, wherein the three antibodies comprise the heavy chain variable region / light chain variable region (HCVR / LCVR) amino acid sequence pairs of (i) SEQ ID NO: 2 / 10, (ii) SEQ ID NO: 20 / 28, and (iii) SEQ ID NO: 38 / 46, and the total antibody content is 100 mg / mL ± 15 mg / mL; (b) 10 mM ± 2 mM histidine buffer, pH 6.0 ± 0.3; (c) 0.1% ± 0.05% w / v polysorbate 80; and (d) 10% ± 2% w / v sucrose. In one embodiment according to this aspect, at least 96% of the antibodies are in the native form after 12 months at 5°C. In one embodiment according to this aspect, at least 97% of the antibodies are in the native form after stirring for 120 minutes. In one embodiment according to this aspect, at least 97% of the antibodies are in the native form after 8 freeze-thaw cycles. In some aspects, the pharmaceutical formulation consists of (a) 100 mg / mL ± 15 mg / mL total antibody, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate 80, and (d) 10% ± 2% w / v sucrose in water at pH 6.0 ± 0.3.

[0052] In one embodiment, the stable liquid formulation contains a total antibody that specifically binds to EBOV at 50 mg / mL ± 7.5 mg / mL and has a viscosity of less than 3 cP at 25°C. In one embodiment, more than 90% of the antibody has a molecular weight of 146 kDa ± 5 kDa, such as 144,804 Da, 145,905 Da, or 143,689 Da. In one embodiment, the pharmaceutical formulation has a viscosity of less than 5 cP, less than 4 cP, or less than 3 cP at 25°C. In one embodiment, at least 90% of the antibody is an undenatured species after 28 days at 45°C. In one embodiment, at least 18% of the antibody is the major charge variant of the antibody after 28 days at 45°C. In one embodiment, at least 96% of the antibody is an undenatured species after 3 months at 25°C. In one embodiment, at least 30% of the antibody is the major charge variant of the antibody after 3 months at 25°C. In one embodiment, at least 96% of the antibody is an undenatured species after 12 months at 5°C. In one embodiment, at least 34% of the antibody is the major charge variant of the antibody after 12 months at 5°C. In one embodiment, at least 97% of the antibody is an undenatured species after 120 minutes of agitation. In one embodiment, at least 35% of the antibody is the major charge variant of the antibody after 120 minutes of agitation. In one embodiment, at least 97% of the antibody is an undenatured species after 8 freeze-thaw cycles. In one embodiment, at least 35% of the antibody is the major charge variant of the antibody after 8 freeze-thaw cycles.

[0053] In one embodiment, the stable liquid formulation contains a total antibody of 100 mg / mL ± 15 mg / mL and has a viscosity of less than 5 cP at 20°C. In one embodiment, more than 90% of the antibody has a molecular weight of 145 kDa ± 2 kDa, for example, 144,804 Da, 145,905 Da, or 143,689 Da. In one embodiment, the pharmaceutical formulation has a viscosity of less than 6 cP, or less than 5 cP, at 20°C. In one embodiment, at least 90% of the antibody is an undenatured species after 28 days at 45°C. In one embodiment, at least 18% of the antibody is the major charge variant of the antibody after 28 days at 45°C. In one embodiment, at least 96% of the antibody is an undenatured species after 3 months at 25°C. In one embodiment, at least 30% of the antibody is the major charge variant of the antibody after 3 months at 25°C. In one embodiment, at least 96% of the antibody is an undenatured species after 12 months at 5°C. In one embodiment, at least 34% of the antibody is the major charge variant of the antibody after 12 months at 5°C. In one embodiment, at least 97% of the antibody is an undenatured species after 120 minutes of agitation. In one embodiment, at least 35% of the antibody is the major charge variant of the antibody after 120 minutes of agitation. In one embodiment, at least 97% of the antibody is an undenatured species after 8 freeze-thaw cycles. In one embodiment, at least 35% of the antibody is the major charge variant of the antibody after 8 freeze-thaw cycles.

[0054] In certain embodiments of this aspect, there is provided a stable liquid formulation comprising: (a) a combination of three antibodies that specifically bind to EBOV, wherein the three antibodies comprise the heavy chain variable region / light chain variable region (HCVR / LCVR) amino acid sequence pairs of (i) SEQ ID NO: 2 / 10, (ii) SEQ ID NO: 20 / 28, and (iii) SEQ ID NO: 38 / 46, and the total antibody content is 50 mg / mL ± 7.5 mg / mL; (b) 10 mM ± 2 mM histidine buffer, pH 6.0 ± 0.3; (c) 0.1% ± 0.05% w / v polysorbate 80; and (d) 10% ± 2% w / v sucrose. In one embodiment of this particular formulation, the viscosity is less than 3 cPoise.

[0055] In certain embodiments of this aspect, there is provided a stable liquid formulation comprising: (a) a combination of three antibodies that specifically bind to EBOV, wherein the three antibodies comprise the heavy chain variable region / light chain variable region (HCVR / LCVR) amino acid sequence pairs of (i) SEQ ID NO: 2 / 10, (ii) SEQ ID NO: 20 / 28, and (iii) SEQ ID NO: 38 / 46, and the total antibody content is 100 mg / mL ± 15 mg / mL; (b) 10 mM ± 2 mM histidine buffer, pH 6.0 ± 0.3; (c) 0.1% ± 0.05% w / v polysorbate 80; and (d) 10% ± 2% w / v sucrose. In one embodiment of this particular formulation, the viscosity is less than 5 cPoise.

[0056] In one aspect, the present disclosure provides a stable liquid pharmaceutical formulation comprising, at pH 6.0 ± 0.3, (a) 5% ± 1% to 15% ± 3% w / v sucrose, (b) 5 mM ± 1 mM to 20 mM ± 4 mM histidine buffer, (c) 0.01% ± 0.005% to 0.5% ± 0.25% w / v polysorbate, and (d) a total anti-EBOV antibody at up to 100 mg / mL ± 15 mg / mL. In another aspect, there is provided a stable liquid pharmaceutical formulation comprising, at pH 6.0 ± 0.3, (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 50 mg / mL ± 7.5 mg / mL total anti-EBOV antibody. The EBOV antibody according to this aspect comprises a combination of three antibodies that specifically bind to EBOV, and the three antibodies comprise the heavy chain variable region / light chain variable region (HCVR / LCVR) amino acid sequence pairs of (i) SEQ ID NO: 2 / 10, (ii) SEQ ID NO: 20 / 28, and (iii) SEQ ID NO: 38 / 46.

[0057] In one aspect, there is provided a stable liquid pharmaceutical formulation comprising, at pH 6.0 ± 0.3, (a) 5% ± 1% to 15% ± 3% w / v sucrose, (b) 5 mM ± 1 mM to 20 mM ± 4 mM histidine buffer, (c) 0.01% ± 0.005% to 0.5% ± 0.25% w / v polysorbate, and (d) 100 mg / mL ± 15 mg / mL total anti-EBOV antibody. In another aspect, there is provided a stable liquid pharmaceutical formulation comprising, at pH 6.0 ± 0.3, (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 100 mg / mL ± 15 mg / mL total antibody. The EBOV antibody according to this aspect comprises a combination of three antibodies that specifically bind to EBOV, and the three antibodies comprise the heavy chain variable region / light chain variable region (HCVR / LCVR) amino acid sequence pairs of (i) SEQ ID NO: 2 / 10, (ii) SEQ ID NO: 20 / 28, and (iii) SEQ ID NO: 38 / 46.

[0058] In one embodiment, the stable liquid formulation contains 150 mg / mL of total anti-EBOV antibody. In one embodiment, the stable liquid formulation contains 100 mg / mL of total anti-EBOV antibody. In one embodiment, the stable liquid formulation contains 50 mg / mL of total anti-EBOV antibody. In one embodiment, the stable liquid formulation contains 10 mM ± 2 mM of histidine buffer. In one embodiment, the stable liquid formulation contains 10% sucrose. In one embodiment, the stable liquid formulation contains 9% sucrose. In one embodiment, the stable liquid formulation contains 8% sucrose. In one embodiment, the stable liquid formulation contains 5% sucrose. In one embodiment, the stable liquid formulation contains 0.1% polysorbate. In one embodiment, the stable liquid formulation contains 0.2% polysorbate. In one embodiment, the polysorbate is polysorbate 80 or polysorbate 20.

[0059] In one embodiment, the stable liquid formulation contains 33.3 mg / mL of an antibody comprising the heavy chain variable region / light chain variable region (HCVR / LCVR) amino acid sequence pair of SEQ ID NO: 2 / 10, 33.3 mg / mL of an antibody comprising the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 20 / 28, and 33.3 mg / mL of an antibody comprising the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 38 / 46 in an aqueous buffer solution containing 10 mM L-histidine, 10% (w / v) sucrose, and 0.1% (w / v) polysorbate 80, pH 6.0. In one embodiment, the stable liquid formulation maintains at least about 90% or at least about 95% of the ADCC potency in an ADCC bioassay after storage at 5°C for 6 months as compared to the same liquid formulation before storage at 5°C for 6 months. In one embodiment, the stable liquid formulation maintains at least about 95% of the pseudovirus neutralization activity after storage at 5°C for 6 months as compared to the same liquid formulation before storage at 5°C for 6 months. In one embodiment, the stable liquid formulation maintains at least about 80% or at least about 85% of the ADCC potency in an ADCC bioassay after storage at 25°C / 60% RH for 6 months as compared to the same liquid formulation before storage at 25°C / 60% RH for 6 months. In one embodiment, the stable liquid formulation maintains at least about 95% of the pseudovirus neutralization activity after storage at 25°C / 60% RH for 6 months as compared to the same liquid formulation before storage at 25°C / 60% RH for 6 months. In one embodiment, the stable liquid formulation maintains at least about 95% of the pseudovirus neutralization activity after storage at 40°C / 75% RH for 6 months as compared to the same liquid formulation before storage at 40°C / 75% RH for 6 months. In one embodiment, the stable liquid formulation maintains at least about 95% of the ADCC potency in an ADCC bioassay after stirring for 120 minutes as compared to the same liquid formulation before stirring for 120 minutes. In one embodiment, the stable liquid formulation maintains at least about 85% of the ADCC potency in an ADCC bioassay after 8 freeze / thaw cycles as compared to the same liquid formulation before 8 freeze / thaw cycles.In one embodiment, the stable liquid formulation maintains at least about 95% of its pseudovirus neutralizing activity after 120 minutes of agitation compared to the same liquid formulation prior to 120 minutes of agitation. In one embodiment, the stable liquid formulation maintains at least about 95% of its pseudovirus neutralizing activity after 8 freeze / thaw cycles compared to the same liquid formulation prior to 8 freeze / thaw cycles.

[0060] In one aspect, the stable liquid pharmaceutical formulation of any of the foregoing aspects is provided in a container. In one embodiment, the container is a vial. In one embodiment, the container is a polycarbonate vial. In one embodiment, the container is a glass vial. In one embodiment, the glass vial is a type 1 clear glass vial. In one embodiment, the glass vial is a type 1 borosilicate glass vial with a fluorocarbon-coated butyl rubber stopper. In one embodiment, the container is a microinjector. In one embodiment, the container is a syringe. In one embodiment, the container is a prefilled syringe. In one embodiment, the syringe contains low tungsten glass. In one embodiment, the syringe includes an autoinjector. In one embodiment, the syringe includes a fluorocarbon-coated plunger. In certain embodiments, the syringe is a 1 mL or 2.25 mL length glass syringe containing less than 500 parts per billion tungsten, with a 27G needle, a fluorocarbon-coated butyl rubber stopper, and a latex-free non-cytotoxic rubber tip cap. In one embodiment, the syringe is a 1 mL length glass syringe with a 27G thin wall needle, a FLUROTEC-coated 4023 / 50 rubber stopper, and an FM 27 rubber tip cap. In one embodiment, the syringe is a 1 mL, 2 mL, 3 mL, 5 mL, or 10 mL plastic syringe with a needle attached.

[0061] In one aspect, there is provided a kit comprising any one of the stable pharmaceutical compositions, containers, and instructions of the foregoing aspects. In one embodiment, the container is a glass vial. In one embodiment, the container is a prefilled syringe. In one embodiment, the container is an autoinjector. In one embodiment, the syringe is a 1 mL or 2.25 mL length glass syringe with a 27G thin wall needle, a FLUROTEC coated 4023 / 50 rubber stopper, and an FM 27 rubber tip cap. In one embodiment, the syringe is a 1 mL, 2 mL, 3 mL, 5 mL, or 10 mL plastic syringe with a needle attached.

[0062] In certain embodiments, the present disclosure provides a prefilled syringe containing a stable liquid pharmaceutical formulation comprising, at pH 6.0 ± 0.3, (i) at least one human antibody that binds specifically to EBOV, at a concentration of 5 ± 0.75 mg / ml to 250 ± 37.5 mg / ml, (ii) a histidine buffer at 5 mM ± 1 mM to 20 ± 4 mM, (iii) polysorbate 80 at 0.05% ± 0.025% to 0.3% ± 0.15% (w / v), and (iv) sucrose at 1% ± 0.2% to 10% ± 2% (w / v), wherein the at least one human anti-EBOV antibody is selected from the group consisting of: (i) a first antibody comprising three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 2 / 10; (ii) a second antibody comprising three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 20 / 28; and (iii) a third antibody comprising three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 38 / 46.In some embodiments, the formulation has a property selected from the group consisting of: at least 90% of the antibody is an undenatured species after 28 days at 45°C; at least 18% of the antibody is the major charge variant of the antibody after 28 days at 45°C; at least 96% of the antibody is an undenatured species after 3 months at 25°C; at least 30% of the antibody is the major charge variant of the antibody after 3 months at 25°C; at least 96% of the antibody is an undenatured species after 12 months at 5°C; at least 34% of the antibody is the major charge variant of the antibody after 12 months at 5°C; at least 97% of the antibody is an undenatured species after 120 minutes of agitation; at least 35% of the antibody is the major charge variant of the antibody after 120 minutes of agitation; at least 97% of the antibody is an undenatured species after 8 freeze-thaw cycles; at least 35% of the antibody is the major charge variant of the antibody after 8 freeze-thaw cycles; more than 90% of the antibody has a molecular weight of 145 kDa ± 2 kDa, for example, a molecular weight of 144,804 Da, 145,905 Da, or 143,689 Da; and the pharmaceutical formulation has a viscosity of less than 20 cP, less than 15 cP, less than 10 cP, less than 5 cP, or less than 3 cP.

[0063] In certain embodiments, the present disclosure provides a glass vial containing a stable liquid pharmaceutical formulation comprising, at pH 6.0 ± 0.3, (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 5 mg / mL ± 0.75 mg / mL to 250 mg / mL ± 37.5 anti-EBOV antibody, wherein at least one anti-EBOV antibody is selected from the group consisting of: (i) a first antibody comprising three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 2 / 10; (ii) a second antibody comprising three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 20 / 28; and (iii) a third antibody comprising three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 38 / 46.In some embodiments, the formulation has a property selected from the group consisting of: the formulation is stable to storage and stress in a glass vial; the formulation is stable and compatible for use in an IV delivery device; the formulation is chemically and physically stable to standard dilutions known in the art (e.g., 0.9% sodium chloride or 5% dextrose or lactated Ringer's, etc.); the formulation is stable to an IV bag made of glass or polymeric plastic (e.g., polyvinyl chloride, phthalate, polyolefin or polypropylene); the formulation is compatible with standard infusion pumps (e.g., peristaltic pump, fluid displacement pump, etc.); at least 90% of the antibody is an undenatured species after 28 days at 45°C; at least 18% of the antibody is the major charge variant of the antibody after 28 days at 45°C; at least 96% of the antibody is an undenatured species after 3 months at 25°C; at least 30% of the antibody is the major charge variant of the antibody after 3 months at 25°C; at least 96% of the antibody is an undenatured species after 12 months at 5°C; at least 34% of the antibody is the major charge variant of the antibody after 12 months at 5°C; at least 97% of the antibody is an undenatured species after 120 minutes of agitation; at least 35% of the antibody is the major charge variant of the antibody after 120 minutes of agitation; at least 97% of the antibody is an undenatured species after 8 freeze-thaw cycles; at least 35% of the antibody is the major charge variant of the antibody after 8 freeze-thaw cycles; more than 90% of the antibody has a molecular weight of 145 kDa ± 2 kDa, e.g., a molecular weight of 144,804 Da, 145,905 Da, or 143,689 Da; and the pharmaceutical formulation has a viscosity of less than 20 cP, less than 15 cP, less than 10 cP, less than 5 cP, or less than 3 cP. In some embodiments, the formulation is stable for transport at a temperature of about 2°C to about 8°C. In some embodiments, the formulation is stable when stored at about 2°C to about 8°C. Stability tests were performed to support the potential for deviation from storage conditions. In some embodiments, the formulation is stable at room temperature for at least about 6 months. In some embodiments, the formulation is physically and chemically stable to agitation and freeze / thaw cycles.In some embodiments, the antibody in the formulation maintains biological activities such as ADCC activity or pseudovirus neutralization activity after exposure to a temperature of up to 25 °C or up to 30 °C, or after agitation or freeze / thaw cycles.

[0064] Other embodiments will become apparent from consideration of the following detailed description of the invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0065]

Figure 1

[0066]

Figure 2

[0067] Detailed Description Therapeutic agents, such as biologic agents, for example, are typically formulated as individual agents in a formulation that provides stability of the therapeutic agent against long-term storage at low temperatures. Packaged biologic agents are handled with care and are often maintained in lyophilized form until just prior to use to minimize aggregation and damage to macromolecules. Lyophilized formulations are typically more stable than liquid formulations. Further, packaged biologic agents are maintained cooled during transport and storage if not frozen.

[0068] Provided herein are stable pharmaceutical formulations comprising antibodies for the treatment of Ebola virus (EBOV) infection or for prophylactic treatment against EBOV exposure. These formulations are stable liquid antibody formulations prepared to withstand rigorous transportation and storage while maintaining stability. The therapeutic agents are prepared in a manner that facilitates their use in remote locations where Ebola hemorrhagic fever outbreaks, such as in Congo or Sudan, are occurring. Thus, the formulations are in liquid form, avoiding the reconstitution step required before using the lyophilized form of the drug and avoiding the potential for contamination of the pharmaceutical. The liquid pharmaceutical formulations are stable even when transported long distances and exposed to stress, such as temperature cycling, extreme temperatures, agitation during transportation, etc., under conditions where the therapeutic agent would be exposed during transportation from the manufacturing facility to remote field clinics where Ebola exposure has occurred and / or where Ebola patients are present.

[0069] In some embodiments, the stable liquid formulation comprises a plurality of antibodies, e.g., a cocktail comprising two or three anti-EBOV antibodies. Stable antibody cocktail formulations are difficult. The protein therapeutics in IV bags are exposed to ambient temperature and visible light for short periods, and thus these mild conditions usually do not induce product degradation. However, during long-term storage, often the individual proteins undergo several degradation processes including enhanced attractive intermolecular protein-protein interactions, increased viscosity, and impairment of structural integrity. The stress conditions present during co-formulation manufacture, transportation, storage, handling, and administration to patients can have an impact and result in important product liabilities such as aggregation, deamidation, oxidation, etc. Furthermore, co-formulation conditions can result in the formation of heterogeneous aggregates. See Svitel et al., BioProcess International, 2019; Patel et al, Journal of Pharmaceutical Sciences, 107:3032-3046, 2018; and Mueller et al., Journal of Pharmacy and Pharmacology, 70:666-674, 2018.

[0070] Before describing the method, it should be understood that the present invention is not limited to the specific methods and experimental conditions described herein, as such methods and conditions may vary. Since the scope of the present disclosure is limited only by the appended claims, it should also be understood that the terms used herein are for the purpose of describing particular embodiments only and are not intended to be limiting.

[0071] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. As used herein, the term "about" when used in reference to a particular recited numerical value or range of values means that the value can vary from the recited value by up to 1% or less. For example, as used herein, the expression "about 100" includes 99 and 101, as well as all values in between (e.g., 99.1, 99.2, 99.3, 99.4, etc.). Any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, although the preferred methods and materials are described hereinafter. All publications mentioned herein are incorporated herein by reference in their entirety.

[0072] As used herein, the expression "pharmaceutical preparation" means a combination of at least one active ingredient (e.g., small molecule, macromolecule, compound, etc. that can exert a biological effect in humans or non-human animals) and at least one inactive ingredient, and the inactive ingredient is suitable for therapeutic administration to humans or non-human animals when combined with the active ingredient or one or more additional inactive ingredients. As used herein, the term "preparation" means "pharmaceutical preparation" unless otherwise indicated. The present disclosure provides a pharmaceutical preparation comprising at least one therapeutic polypeptide. According to a particular embodiment of the present disclosure, the therapeutic polypeptide is an antibody or an antigen-binding fragment thereof that specifically binds to Ebola virus (EBOV). More specifically, the present disclosure includes a stable pharmaceutical preparation comprising (i) one or more human antibodies that specifically bind to EBOV, (ii) a histidine buffer, (iii) an organic co-solvent that is a non-ionic surfactant, and (iv) a stabilizer that is a carbohydrate. In one particular embodiment, the stable pharmaceutical preparation comprises (i) three human antibodies that specifically bind to EBOV, (ii) a histidine buffer, (iii) an organic co-solvent that is a non-ionic surfactant, and (iv) a stabilizer that is a carbohydrate. Specific exemplary components and preparations included in the present disclosure are described in detail below.

[0073] Antibody that specifically binds to EBOV The pharmaceutical preparation of the present disclosure may comprise a human antibody that specifically binds to EBOV or an antigen-binding fragment thereof. As used herein, the term "EBOV" means Ebola virus. Antibodies against EBOV are described, for example, in U.S. Patent / Publication Nos. 10,501,526, 10,081,670, 9,771,414, 6,630,144, 6,875,433, 7,335,356 and 8,513,391, and WO2016 / 123019, EP1539238, EP2350270 and EP8513391.

[0074] As used herein, the term "antibody" generally refers to an immunoglobulin molecule containing four polypeptide chains, two heavy (H) chains and two light (L) chains, and multimers thereof (e.g., IgM), the chains being interconnected by disulfide bonds. However, immunoglobulin molecules consisting of only heavy chains (i.e., lacking light chains) are also included within the definition of the term "antibody". Each heavy chain includes a heavy chain variable region (referred to herein as HCVR or V H which is omitted) and a heavy chain constant region. The heavy chain constant region includes three domains, CH1, CH2, and CH3. Each light chain includes a light chain variable region (referred to herein as LCVR or V L which is omitted) and a light chain constant region. The light chain constant region includes one domain (CL1). The V H region and the V L region can be further subdivided into hypervariable regions called complementarity determining regions (CDRs) interspersed with more conserved regions called framework regions (FRs). Each V H and V L is composed of three CDRs and four FRs arranged from the amino terminus to the carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.

[0075] Unless otherwise specifically indicated, as used herein, the term "antibody" should be understood to encompass whole antibody molecules as well as antigen-binding fragments thereof. The term "antigen-binding portion" or "antigen-binding fragment" of an antibody (or simply "antibody portion" or "antibody fragment"), as used herein, refers to one or more fragments of an antibody that possess the ability to specifically bind to EBOV or its epitope.

[0076] As used herein, "isolated antibody" is intended to refer to an antibody that is substantially free of other antibodies having different antigen specificities (e.g., an isolated antibody that specifically binds to EBOV is substantially free of antibodies that specifically bind to antigens other than EBOV).

[0077] Terms such as "specifically binds" mean that an antibody or its antigen-binding fragment forms a complex with an antigen that is relatively stable under physiological conditions. Specific binding can be characterized by a dissociation constant of at least about 1×10 -8 M or greater. Methods for determining whether two molecules specifically bind are well known in the art and include, for example, equilibrium dialysis, surface plasmon resonance, and the like. In the context of the present disclosure, multispecific (e.g., bispecific) antibodies that bind to EBOV as well as one or more additional antigens are considered to "specifically bind" to EBOV. Further, an isolated antibody may be substantially free of other cellular materials and / or chemical substances.

[0078] Examples of anti-EBOV antibodies that can be included in the pharmaceutical formulations of the present disclosure are described in Patent Application Publication US 2016 / 0215040 and WO 2016 / 123019, the disclosures of which are incorporated herein by reference in their entirety. Of particular interest are the three antibodies, H1H17203P, H1H17139P, and H1H17161P, which are formulated independently or in any combination in stable pharmaceutical formulations disclosed therein.

[0079] According to certain embodiments of the present disclosure, an anti-EBOV antibody or its antigen-binding fragment comprises heavy chain complementarity determining region (HCDR) 1 of SEQ ID NO: 4, HCDR2 of SEQ ID NO: 6, and HCDR3 of SEQ ID NO: 8. In certain embodiments, the anti-EBOV antibody or its antigen-binding fragment comprises the HCVR of SEQ ID NO: 2.

[0080] According to certain embodiments of the present disclosure, an anti-EBOV antibody or its antigen-binding fragment comprises light chain complementarity determining region (LCDR) 1 of SEQ ID NO: 12, LCDR2 of SEQ ID NO: 14, and LCDR3 of SEQ ID NO: 16. In certain embodiments, the anti-EBOV antibody or its antigen-binding fragment comprises the LCVR of SEQ ID NO: 10.

[0081] According to certain embodiments of the present disclosure, the anti-EBOV antibody or antigen-binding fragment thereof comprises an HCVR having 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 2.

[0082] According to certain embodiments of the present disclosure, the anti-EBOV antibody or antigen-binding fragment thereof comprises an LCVR having 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 10.

[0083] According to certain embodiments of the present disclosure, the anti-EBOV antibody or antigen-binding fragment thereof comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 2 having five or fewer amino acid substitutions.

[0084] According to certain embodiments of the present disclosure, the anti-EBOV antibody or antigen-binding fragment thereof comprises an LCVR comprising the amino acid sequence of SEQ ID NO: 10 having two or fewer amino acid substitutions.

[0085] According to certain embodiments of the present disclosure, the anti-EBOV antibody or antigen-binding fragment thereof comprises heavy chain complementarity determining region (HCDR) 1 of SEQ ID NO: 22, HCDR2 of SEQ ID NO: 24, and HCDR3 of SEQ ID NO: 26. In certain embodiments, the anti-EBOV antibody or antigen-binding fragment thereof comprises an HCVR of SEQ ID NO: 20.

[0086] According to certain embodiments of the present disclosure, the anti-EBOV antibody or antigen-binding fragment thereof comprises light chain complementarity determining region (LCDR) 1 of SEQ ID NO: 30, LCDR2 of SEQ ID NO: 32, and LCDR3 of SEQ ID NO: 34. In certain embodiments, the anti-EBOV antibody or antigen-binding fragment thereof comprises an LCVR of SEQ ID NO: 28.

[0087] According to certain embodiments of the present disclosure, the anti-EBOV antibody or antigen-binding fragment thereof comprises an HCVR having 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 20.

[0088] According to certain embodiments of the present disclosure, an anti-EBOV antibody or an antigen-binding fragment thereof comprises an LCVR having 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 28.

[0089] According to certain embodiments of the present disclosure, an anti-EBOV antibody or an antigen-binding fragment thereof comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 20 having five or fewer amino acid substitutions.

[0090] According to certain embodiments of the present disclosure, an anti-EBOV antibody or an antigen-binding fragment thereof comprises an LCVR comprising the amino acid sequence of SEQ ID NO: 28 having two or fewer amino acid substitutions.

[0091] According to certain embodiments of the present disclosure, an anti-EBOV antibody or an antigen-binding fragment thereof comprises heavy chain complementarity determining region (HCDR) 1 of SEQ ID NO: 40, HCDR2 of SEQ ID NO: 42, and HCDR3 of SEQ ID NO: 44. In certain embodiments, the anti-EBOV antibody or an antigen-binding fragment thereof comprises an HCVR of SEQ ID NO: 38.

[0092] According to certain embodiments of the present disclosure, an anti-EBOV antibody or an antigen-binding fragment thereof comprises light chain complementarity determining region (LCDR) 1 of SEQ ID NO: 48, LCDR2 of SEQ ID NO: 50, and LCDR3 of SEQ ID NO: 52. In certain embodiments, the anti-EBOV antibody or an antigen-binding fragment thereof comprises an LCVR of SEQ ID NO: 46.

[0093] According to certain embodiments of the present disclosure, an anti-EBOV antibody or an antigen-binding fragment thereof comprises an HCVR having 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 38.

[0094] According to certain embodiments of the present disclosure, an anti-EBOV antibody or an antigen-binding fragment thereof comprises an LCVR having 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 46.

[0095] According to certain embodiments of the present disclosure, an anti-EBOV antibody or an antigen-binding fragment thereof comprises a HCVR comprising the amino acid sequence of SEQ ID NO: 38 having five or fewer amino acid substitutions.

[0096] According to certain embodiments of the present disclosure, an anti-EBOV antibody or an antigen-binding fragment thereof comprises a LCVR comprising the amino acid sequence of SEQ ID NO: 46 having two or fewer amino acid substitutions.

[0097] According to certain embodiments of the present disclosure, a stable liquid pharmaceutical formulation comprises one or more of the above-described anti-EBOV antibodies or antigen-binding fragments thereof.

[0098] Sequence identity can be measured by any method known in the art (e.g., GAP, BESTFIT, and BLAST).

[0099] The present disclosure also includes a stable liquid formulation comprising an anti-EBOV antibody, wherein the anti-EBOV antibody comprises a variant of any of the amino acid sequences of the HCVR, LCVR, and / or CDR disclosed herein having one or more amino acid substitutions, such as conservative amino acid substitutions. Illustratively, the present disclosure includes a formulation comprising an anti-EBOV antibody having an amino acid sequence of a HCVR, LCVR, and / or CDR having amino acid substitutions, such as conservative amino acid substitutions, of, for example, 10 or fewer, 8 or fewer, 6 or fewer, 4 or fewer, etc., relative to any of the amino acid sequences of the HCVR, LCVR, and / or CDR disclosed herein.

[0100] In certain embodiments, the anti-EBOV antibody comprises an Fc region selected from the group consisting of human IgG1, IgG2, IgG3, and IgG4 isotypes.

[0101] According to certain embodiments of the present disclosure, an anti-EBOV antibody or an antigen-binding fragment thereof comprises a heavy chain of SEQ ID NO: 17 and a light chain of SEQ ID NO: 18.

[0102] According to certain embodiments of the present disclosure, an anti-EBOV antibody or an antigen-binding fragment thereof comprises a heavy chain of SEQ ID NO: 35 and a light chain of SEQ ID NO: 36.

[0103] According to certain embodiments of the present disclosure, an anti-EBOV antibody or an antigen-binding fragment thereof comprises a heavy chain of SEQ ID NO: 53 and a light chain of SEQ ID NO: 54.

[0104] It is known in the art that amino acid terminal truncations can occur during antibody production (see, e.g., Wang et al 2007, J. Pharma. Sci. 96:1-26). Thus, in certain embodiments, the anti-EBOV antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 17, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 35, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 53. In some aspects, the heavy chain lacks the C-terminal lysine from the amino acid sequence of SEQ ID NO: 17. In some aspects, the heavy chain lacks the C-terminal lysine from the amino acid sequence of SEQ ID NO: 35. In some aspects, the heavy chain amino lacks the C-terminal lysine from the amino acid sequence of SEQ ID NO: 53. In certain embodiments, the formulations of the present disclosure contain about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, about 98%, or more of the anti-EBOV antibody, wherein the C-terminal lysine is absent.

[0105] The amount of antibody, combination of antibodies, or antigen-binding fragment thereof contained in the stable liquid pharmaceutical formulations of the present disclosure can vary depending on the desired specific properties of the formulation, as well as the particular circumstances and the intended purpose for which the formulation is to be used. For example, it may be necessary to transport a formulation containing one or more anti-EBOV antibodies from the manufacturing site to remote regions of the world, such as the Congo or other African regions, where the formulation may be exposed to agitation during transit and / or to elevated temperatures during transit or at the point of care.

[0106] In certain embodiments, the pharmaceutical formulation is a stable liquid formulation that may contain a total antibody of 5 ± 0.75 mg / mL to 250 ± 37.5 mg / mL; 10 ± 1.5 mg / mL to 240 ± 36 mg / mL; 20 ± 3.0 mg / mL to 230 ± 34.5 mg / mL; 25 ± 3.75 mg / mL to 240 ± 36 mg / mL; 50 ± 7.5 mg / mL to 230 ± 34.5 mg / mL; 60 ± 9 mg / mL to 240 ± 36 mg / mL; 70 ± 10.5 mg / mL to 230 ± 34.5 mg / mL; 80 ± 12 mg / mL to 220 ± 33 mg / mL; 90 ± 13.5 mg / mL to 210 ± 31.5 mg / mL; 100 ± 15 mg / mL to 200 ± 30 mg / mL; 110 ± 16.5 mg / mL to 190 ± 28.5 mg / mL; 120 ± 18 mg / mL to 180 ± 27 mg / mL; 130 ± 19.5 mg / mL to 170 ± 25.5 mg / mL; 140 ± 21 mg / mL to 160 ± 24 mg / mL; 150 ± 22.5 mg / mL; or 175 ± 26.25 mg / mL.For example, the formulations of the present disclosure can contain total antibodies or antigen-binding fragments thereof at about 5 mg / mL; about 10 mg / mL; about 15 mg / mL; about 20 mg / mL; about 25 mg / mL; about 30 mg / mL; about 35 mg / mL; about 40 mg / mL; about 45 mg / mL; about 50 mg / mL; about 55 mg / mL; about 60 mg / mL; about 65 mg / mL; about 70 mg / mL; about 75 mg / mL; about 80 mg / mL; about 85 mg / mL; about 90 mg / mL; about 95 mg / mL; about 100 mg / mL; about 105 mg / mL; about 110 mg / mL; about 115 mg / mL; about 120 mg / mL; about 125 mg / mL; about 130 mg / mL; about 135 mg / mL; about 140 mg / mL; about 145 mg / mL; about 150 mg / mL; about 155 mg / mL; about 160 mg / mL; about 165 mg / mL; about 170 mg / mL; about 175 mg / mL; about 180 mg / mL; about 185 mg / mL; about 190 mg / mL; about 195 mg / mL; about 200 mg / mL; about 205 mg / mL; about 210 mg / mL; about 215 mg / mL; about 220 mg / mL; about 225 mg / mL; about 230 mg / mL; about 235 mg / mL; about 240 mg / mL; about 245 mg / mL; or about 250 mg / mL that specifically bind to EBOV.

[0107] Excipients and pH The pharmaceutical formulations of the present disclosure contain one or more excipients. As used herein, the term "excipient" means any non-therapeutic agent added to a formulation to provide desirable consistency, viscosity, or stabilizing effects.

[0108] In certain embodiments, the pharmaceutical formulations disclosed herein include a stabilizing type and amount of at least one organic co-solvent, such as vortexing, cyclic vibration, and under harsh handling and agitation conditions such as shock, to stabilize EBOV. In some embodiments, "stabilizing" means preventing the formation of aggregated antibodies, such as preventing the formation of aggregated antibodies that exceed 0% of the total amount of antibodies, or the formation of aggregated antibodies that exceed 1%, or the formation of aggregated antibodies that exceed 3% during the process of harsh handling. In some embodiments, the co-solvent stabilizes the formulation to prevent the formation of 0% to 3% aggregated antibodies. In some embodiments, the co-solvent stabilizes the formulation to prevent the formation of aggregated antibodies that exceed 0%. In some embodiments, the harsh handling is to vortex a solution containing the antibody and the organic co-solvent for about 60 minutes or about 120 minutes.

[0109] In certain embodiments, the organic co-solvent is a non-ionic surfactant such as alkyl poly(ethylene oxide). Specific non-ionic surfactants that can be included in the formulations of the present disclosure include, for example, polysorbates such as polysorbate 20, polysorbate 28, polysorbate 40, polysorbate 60, polysorbate 65, polysorbate 80, polysorbate 81, and polysorbate 85; poloxamers such as poloxamer 181, poloxamer 188, poloxamer 407; or polyethylene glycol (PEG). Polysorbate 20 is also known as TWEEN20, sorbitan monolaurate, and polyoxyethylene sorbitan monolaurate. Poloxamer 188 is also known as PLURONIC F68.

[0110] The amount of nonionic surfactant contained in the pharmaceutical formulations of the present disclosure can vary depending on the desired specific properties of the formulation, as well as the particular circumstances and the intended purpose for which the formulation is to be used. In certain embodiments, the formulation may contain from 0.01% ± 0.005% to 0.5% ± 0.25% surfactant. For example, the formulations of the present disclosure may contain about 0.005%, about 0.01%, about 0.02%, about 0.03%, about 0.04%, about 0.05%, about 0.06%, about 0.07%, about 0.08%, about 0.09%, about 0.1%, about 0.11%, about 0.12%, about 0.13%, about 0.14%, about 0.15%, about 0.16%, about 0.17%, about 0.18%, about 0.19%, about 0.20%, about 0.21%, about 0.22%, about 0.23%, about 0.24%, about 0.25%, about 0.26%, about 0.27%, about 0.28%, about 0.29%, about 0.30%, about 0.35%, about 0.40%, about 0.45%, about 0.46%, about 0.47%, about 0.48%, about 0.49%, about 0.50%, about 0.55%, or about 0.575% of polysorbate 20 or polysorbate 80.

[0111] Furthermore, the pharmaceutical formulations of the present disclosure may contain one or more stabilizers of a type and in an amount that stabilizes the EBOV antibody under heat stress conditions. In some embodiments, “stabilize” means that when a solution containing the antibody and the heat stabilizer is maintained at about 45° C. for at least about 28 days, more than about 91% of the antibody is maintained in an unfolded conformation. In some embodiments, “stabilize” means that when a solution containing the antibody and the heat stabilizer is maintained at about 45° C. for at least about 28 days, less than about 6% of the antibody aggregates. As used herein, “unfolded” means the major form of the antibody by size exclusion and is generally the intact monomer of the antibody. The term “unfolded” also refers to the non-aggregated and non-degraded forms of the antibody.

[0112] In some embodiments, the heat stabilizer is a polyol. In some embodiments, the heat stabilizer is an amino acid. In some aspects, the heat stabilizer is a sugar such as, for example, sucrose. The amount of stabilizer contained in the formulation can vary depending on the particular circumstances in which the formulation is used and the intended purpose. In certain embodiments, the formulation may contain from about 1% to about 15% sugar, from about 2% to about 14% sugar, from about 3% to about 13% sugar, from about 4% to about 12% sugar, from about 5% to about 12% sugar, from about 6% to about 11% sugar, from about 7% to about 10% sugar, from about 8% to about 11% sugar, or from about 9% to about 11% sugar. For example, the pharmaceutical formulations of the present disclosure may contain 4% ± 0.8%, 5% ± 1%, 6% ± 1.2%, 7% ± 1.4%, 8% ± 1.6%, 9% ± 1.8%, 10% ± 2%, 11% ± 2.2%, 12% ± 2.4%, 13% ± 2.6%, or about 14% ± 2.8% sugar (e.g., sucrose).

[0113] The pharmaceutical formulations of the present disclosure may also contain a buffer or buffer system that helps maintain a stable pH and aids in the stabilization of EBOV antibodies. As used herein, the term "buffer" refers to a pharmaceutically acceptable buffer that maintains a stable pH or resists changes in the pH of a solution. In a preferred embodiment, the buffer contains histidine. In the context of the present disclosure, a "histidine buffer" or "buffer containing histidine" is a buffer that contains the amino acid histidine. Examples of histidine buffers include histidine chloride, histidine acetate, histidine phosphate, and histidine sulfate. In one embodiment, the histidine buffer is prepared by dissolving L-histidine and L-histidine hydrochloride (e.g., as the monohydrate) in defined amounts and ratios. In one embodiment, the histidine buffer is prepared by titrating L-histidine (free base, solid) with dilute hydrochloric acid. The term "histidine" is used interchangeably with "histidine buffer" throughout the present disclosure. In some embodiments, "stabilize" means that less than 10% ± 0.5% of the antibody aggregates when a solution containing the antibody and buffer is maintained at about 45°C for at least about 28 days. In some embodiments, "stabilize" means that less than 5% ± 0.5% or less than 4% ± 0.5% of the antibody aggregates when a solution containing the antibody and buffer is maintained at about 25°C for up to about 3 months. In some embodiments, "stabilize" means that less than 5% ± 0.5% or less than 4% ± 0.5% of the antibody aggregates when a solution containing the antibody and buffer is maintained at about 5°C for up to about 36 months. In some embodiments, "stabilize" means that at least 90% ± 0.5% or at least 94% ± 0.5% of the antibody is in its native conformation as determined by size exclusion chromatography when a solution containing the antibody and buffer is maintained at about 45°C for at least about 28 days.In some embodiments, "stabilizing" means that when a solution containing an antibody and a buffer is maintained at about 25°C for up to about 3 months, at least 95% ± 0.5% or at least 96% ± 0.5% of the antibody is in its native conformation as determined by size exclusion chromatography. In some embodiments, "stabilizing" means that when a solution containing an antibody and a buffer is maintained at about 5°C for up to about 12 months, at least 95% ± 0.5% or at least 96% ± 0.5% of the antibody is in its native conformation as determined by size exclusion chromatography. "Native" or "native conformation" means an antibody fraction that is not aggregated or degraded. This is generally determined by an assay that measures the relative size of the antibody entity, such as a size exclusion chromatography assay. Non-aggregated and non-degraded antibodies elute in a fraction equivalent to the native antibody and are generally the major elution fraction. Aggregated antibodies elute in a fraction that exhibits a larger size than the native antibody. Degraded antibodies elute in a fraction that exhibits a smaller size than the native antibody.

[0114] In some embodiments, "stabilizing" means that when a solution containing an antibody and a buffer is maintained at about 45°C for at least about 28 days, or at least about 1 month, at least 18% ± 0.5% of the antibody is in its major charge form as determined by cation exchange chromatography. In some embodiments, "stabilizing" means that when a solution containing an antibody and a buffer is maintained at about 25°C for up to about 3 months, at least 30% ± 0.5% or at least 35% ± 0.5% of the antibody is in its major charge form as determined by cation exchange chromatography. In some embodiments, "stabilizing" means that when a solution containing an antibody and a buffer is maintained at about 5°C for up to about 12 months, at least 30% ± 0.5% or at least 34% ± 0.5% of the antibody is in its major charge form as determined by cation exchange chromatography. "Major charge" or "major charge form" means the fraction of the antibody that elutes from the ion exchange resin at the major peak, and generally is adjacent to a more "basic" peak on one side and a more "acidic" peak on the other side.

[0115] The pharmaceutical formulation of the present disclosure may have a pH of from about 5.2 to about 6.4. For example, the formulation of the present disclosure may have a pH of about 5.5, about 5.6, about 5.7, about 5.8, about 5.9, about 6.0, about 6.1, about 6.2, about 6.3, about 6.4, or about 6.5. In some embodiments, the pH is 6.0 ± 0.4, 6.0 ± 0.3, 6.0 ± 0.2, 6.0 ± 0.1, about 6.0, or 6.0.

[0116] In some embodiments, the buffer or buffer system comprises at least one buffer having a buffering range that completely or partially overlaps with the range of pH 5.5 to 7.4. In certain embodiments, the buffer comprises a histidine buffer. In certain embodiments, the histidine buffer is present at a concentration of 5 mM ± 1 mM to 15 mM ± 3 mM, 6 mM ± 1.2 mM to 14 mM ± 2.8 mM, 7 mM ± 1.4 mM to 13 mM ± 2.6 mM, 8 mM ± 1.6 mM to 12 mM ± 2.4 mM, 9 mM ± 1.8 mM to 11 mM ± 2.2 mM, 10 mM ± 2 mM, or about 10 mM. In certain embodiments, the buffer system comprises 10 mM ± 2 mM of histidine at pH 6.0 ± 0.3. In certain embodiments, the histidine buffer comprises L-histidine and L-histidine monohydrochloride monohydrate. In one embodiment, the histidine buffer comprises L-histidine at a concentration of 4.8 mM ± 0.96 mM. In one embodiment, the histidine buffer comprises L-histidine monohydrochloride monohydrate at a concentration of 5.2 mM ± 1.04 mM. In one embodiment, the histidine buffer comprises L-histidine at a concentration of 4.8 mM ± 0.96 mM and L-histidine monohydrochloride monohydrate at a concentration of 5.2 mM ± 1.04 mM.

[0117] The pharmaceutical formulations of the present disclosure may contain one or more excipients that serve to maintain the low viscosity of formulations containing a high concentration of anti-EBOV antibody drug substance (e.g., generally an antibody of 150 mg / ml or more) or to reduce the viscosity. In certain embodiments, the viscosity modifier is an amino acid such as, for example, proline or histidine.

[0118] During the antibody purification process, it may be desirable or necessary to exchange one buffer with another to achieve appropriate excipient concentrations, antibody concentrations, pH, etc. Buffer exchange can be achieved, for example, by performing ultrafiltration / diafiltration (UF / DF) using a semi-permeable tangential flow filtration membrane. However, using such techniques can cause the Gibbs-Donnan effect [Bolton et al., 2011, Biotechnol. Prog. 27(1):140-152]. During protein concentration, the positive charge increases on the product side of the membrane, which is electrically neutralized by the selective movement of positive ions to the opposite side of the membrane. A potential consequence of this phenomenon is that the final concentration of certain components (e.g., histidine) is lower than the target concentration of these components, due to the electrostatic repulsion of the positively charged diafiltration buffer excipient for the positively charged antibody protein during the UF / DF process. Accordingly, the present disclosure includes formulations in which the concentration of, for example, histidine, differs from the amounts or ranges recited herein due to the Gibbs-Donnan effect.

[0119] Steric exclusion accounts for the behavior of high-concentration samples. In high-concentration samples, most of the total volume of the solution is taken up by solutes, particularly macromolecules such as proteins, for example, and the co-solvent is excluded from this space. The total volume of co-solvent available for dissolving other solutes then decreases, resulting in a non-uniform distribution across the ultrafiltration membrane. Accordingly, the present disclosure includes formulations in which the concentration of, for example, histidine, may differ from the amounts or ranges recited herein due to the steric exclusion effect.

[0120] During the manufacture of the formulations of the present disclosure, variations may occur in the composition of the formulations. These variations can include the concentration of the active ingredient, the concentration of the excipient, and / or the pH of the formulation. A change in any of these parameters can potentially affect the stability or efficacy of the pharmaceutical, so a structural stability test of the formulation was conducted to evaluate whether variations in the composition within the specified range affect the stability or efficacy of the antibody. Accordingly, the present disclosure includes formulations containing anti-EBOV antibodies that are stable and retain efficacy with up to a 50% variation in excipient concentration. For example, described herein are anti-EBOV antibody formulations in which the stability and efficacy of the formulation are not affected by variations of ±10%, ±20%, ±30%, ±40% or ±50% in the concentration of the antibody, sucrose, histidine buffer and / or polysorbate.

[0121] Stability and Viscosity of Pharmaceutical Formulations As shown in the following examples, the inventors have made the surprising discovery that stable liquid formulations containing high concentrations of one or more anti-EBOV antibodies (e.g., about 50 mg / mL or about 100 mg / mL) can be obtained by formulating the antibody with about 0.1% polysorbate 80, about 10% sucrose, and about 10 mM histidine buffer. In some embodiments, the stable liquid formulation contains three anti-EBOV antibodies at a total antibody concentration of 50 mg / mL or 100 mg / mL, 0.1% polysorbate 80, 10% sucrose, and about 10 mM histidine buffer at a pH of about 6. Such formulations are stable against the stress of rough handling, even when containing three different antibodies, and are stable for storage in the temperature range of -80°C to 45°C, e.g., -30°C, -20°C, 5°C, 25°C (as shown herein), and have a low viscosity (having a viscosity of less than 5 cP).

[0122] The pharmaceutical formulations of the present disclosure typically exhibit a high level of stability. As used herein in connection with pharmaceutical formulations, the term "stable" means that the antibody within the pharmaceutical formulation retains an acceptable degree of chemical structure or biological function after storage under defined conditions. A formulation can be stable even if the antibody contained in the formulation does not maintain 100% of its chemical structure or biological function after storage for a defined period of time. Under certain circumstances, a formulation may be considered "stable" if about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% of the structure or function of the antibody is maintained after storage for a defined period of time.

[0123] Stability can be measured, in particular, by determining the proportion of the non-modified antibody remaining in the formulation after storage at a specified temperature for a specified time. The proportion of the non-modified antibody can be determined, in particular, by size exclusion chromatography (e.g., size exclusion ultra performance liquid chromatography [SE-UPLC]), and non-modified means not aggregated and not degraded. "Acceptable level of stability", when the term is used herein, means that at least 90% of the antibody in the non-modified form can be detected in the formulation after storage at a given temperature for a specified time. In certain embodiments, at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% of the antibody in the non-modified form can be detected in the formulation after storage at a specified temperature for a specified time. The specified time after which stability is measured can be at least 14 days, at least 28 days, at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 12 months, at least 18 months, at least 24 months, or longer. The specified temperature at which the pharmaceutical formulation can be stored when evaluating stability can be any temperature from about -80°C to about 60°C, for example, storage at about -80°C, about -30°C, about -20°C, about 0°C, about 4°C to 8°C, about 5°C, about 25°C, about 35°C, about 37°C, about 40°C, or about 45°C. For example, if after storage at 40°C / 75% relative humidity (RH) for 28 days, more than about 95%, 96%, 97%, or 98% of the non-modified antibody is detected by SE-UPLC, the pharmaceutical formulation can be considered stable. If after storage at 5°C for 12 months, more than about 95%, 96%, 97%, or 98% of the non-modified antibody is detected by SE-UPLC, the pharmaceutical formulation can be considered stable. Even if after storage at 25°C for 3 months, more than about 95%, 96%, 97%, or 98% of the non-modified antibody is detected by SE-UPLC, the pharmaceutical formulation can be considered stable. Also, the pharmaceutical formulation can be considered stable if after storage at 45°C for 28 days, more than about 89%, 90%, 91%, 92%, 93%, 94%, 95% or 96% of the non-modified antibody is detected by SE-UPLC.Even if more than about 96%, 97%, or 98% of the non-denatured antibody is detected by SE-UPLC after storage at -20°C for 12 months, the pharmaceutical preparation may be considered stable. Even if more than about 96%, 97%, or 98% of the non-denatured antibody is detected by SE-UPLC after storage at -30°C for 12 months, the pharmaceutical preparation may be considered stable. Even if more than about 96%, 97%, or 98% of the non-denatured antibody is detected by SE-UPLC after storage at -80°C for 12 months, the pharmaceutical preparation may be considered stable.

[0124] Stability can be measured, in particular, by determining the proportion of antibody formed as aggregates within the formulation after storage at a defined temperature for a defined time, and stability is inversely proportional to the proportion of aggregates formed. The proportion of aggregated antibody can be determined, in particular, by size exclusion chromatography (e.g., size exclusion ultra performance liquid chromatography [SE-UPLC]). "Acceptable stability" when used in this specification means that after storage at a given temperature for a defined time, up to 5% of the antibody in the formulation is detected as the aggregated form (also shown as the high molecular weight - HMW - form). In certain embodiments, acceptable stability means that after storage at a given temperature for a defined time, up to about 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the antibody can be detected as aggregates in the formulation. The defined time after which stability is measured can be at least 2 weeks, at least 28 days, at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 12 months, at least 18 months, at least 24 months, or more. The temperature at which the pharmaceutical formulation can be stored when evaluating stability can be any temperature from about -80°C to about 45°C, for example, storage at about -80°C, about -30°C, about -20°C, about 0°C, about 4°C to 8°C, about 5°C, about 25°C, about 35°C, about 37°C, about 40°C, or about 45°C. For example, if after storage at 5°C for 12 months, less than about 2%, 1%, 0.5%, or 0.1% of the antibody is detected as the aggregated form, the pharmaceutical formulation can be considered stable. Even if after storage at 25°C for 3 months, less than about 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the antibody is detected as the aggregated form, the pharmaceutical formulation can be considered stable. Even if after storage at 40°C / 75%RH for 28 days, less than about 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the antibody is detected as the aggregated form, the pharmaceutical formulation can be considered stable. Even if after storage at 45°C for 28 days, less than about 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or 0.5% of the antibody is detected as the aggregated form, the pharmaceutical formulation can be considered stable.Even if less than about 3%, 2%, 1%, 0.5%, or 0.1% of the antibody is detected in the aggregated form after storage at -20 °C, -30 °C, or -80 °C for 3 months, the pharmaceutical preparation can be considered stable.

[0125] Stability can be measured, in particular, by determining the proportion of antibodies (acidic forms) that migrate into more acidic fractions rather than in the major antibody fraction (major charge form) during ion exchange, and stability is inversely proportional to the fraction of acidic form antibodies. Without wishing to be bound by theory, deamidation of an antibody can make the antibody more negatively charged and thus more acidic compared to the non-deamidated antibody (see, for example, Robinson, N., Protein Deamidation, PNAS, April 16, 2002, 99(8):5283-5288). The proportion of “acidic” antibodies can be determined, in particular, by ion exchange chromatography (e.g., cation exchange ultra-high performance liquid chromatography [CEX-UPLC]). “Acceptable stability” as used herein means that after storage at a defined temperature for a defined time, up to 45% of the antibody in the formulation is detected in the more acidic form. In certain embodiments, acceptable stability means that after storage at a given temperature for a defined time, up to about 40%, 35%, 30%, 25%, 20%, 15%, 10%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the antibody in the formulation can be detected in the acidic form. In one embodiment, acceptable stability means that after storage at a given temperature for a defined time, less than 30%, 25%, 20%, 15%, 10%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the antibody in the formulation can be detected in the acidic form. The defined time after which stability is measured can be at least 2 weeks, at least 28 days, at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 12 months, at least 18 months, at least 24 months, or more. The temperature at which the pharmaceutical formulation can be stored when evaluating stability can be any temperature from about -80°C to about 45°C, for example, storage at about -80°C, about -30°C, about -20°C, about 0°C, about 4°C - 8°C, about 5°C, about 25°C, or about 45°C.For example, a pharmaceutical preparation may be considered stable if less than about 30%, 29%, 28%, 27%, 26%, 25%, 24%, 23%, 22%, 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the antibody is in a more acidic form after storage at -80°C, -30°C, or -20°C for 3 months. The pharmaceutical preparation may be considered stable if less than about 32%, 31%, 30%, 29%, 28%, 27%, 26%, 25%, 24%, 23%, 22%, 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the antibody is in a more acidic form even after storage at 5°C for 12 months. The pharmaceutical preparation may be considered stable if less than about 43%, 42%, 41%, 40%, 39%, 38%, 37%, 36%, 35%, 34%, 33%, 32%, 31%, 30%, 29%, 28%, 27%, 26%, 25%, 24%, 23%, 22%, 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the antibody is in a more acidic form even after storage at 25°C for 3 months. The pharmaceutical preparation may be considered stable if less than about 49%, 48%, 47%, 46%, 45%, 44%, 43%, 42%, 41%, 40%, 39%, 38%, 37%, 36%, 35%, 34%, 33%, 32%, 31%, 30%, 29%, 28%, 27%, 26%, 25%, 24%, 23%, 22%, 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the antibody is detected in a more acidic form even after storage at 45°C for 28 days.

[0126] For example, the stability of the formulations of the present disclosure may be evaluated using other methods such as differential scanning calorimetry (DSC) to determine thermal stability, agitation control to determine mechanical stability, and absorbance at about 350 nm or about 405 nm to determine solution turbidity. For example, the formulations of the present disclosure, after storage for 6 months or more at about 5°C to about 25°C, the OD 405 change of the formulation is less than about 0.05 (e.g., 0.04, 0.03, 0.02, 0.01 or less) from the OD 405 of the formulation at zero time, it may be considered stable.

[0127] The stability may be evaluated using the measurement of the biological activity or binding affinity of the antibody to the target. For example, the formulations of the present disclosure, after storage for a specified time (e.g., 1 to 36 months) at 5°C, 25°C, 45°C, etc., if the anti-EBOV antibody contained in the formulation is at least 90%, 95% or more of the binding affinity of the antibody before such storage, it may be considered stable. The binding affinity can be determined by, for example, ELISA or surface plasmon resonance. The biological activity can be determined by an assay of EBOV activity, such as contacting cells infected with EBOV with a formulation containing an anti-EBOV antibody. The binding of the antibody to such cells can be directly measured, for example, via FACS analysis. Alternatively, the activity of the antibody can be determined by a decrease in the amount of virus in vitro or in vivo, or by an increase in the survival of mammals infected with EBOV.

[0128] Furthermore, the stability can be evaluated by measuring the relative potency of the antibody in an antibody-dependent cell-mediated cytotoxicity assay (ADCC assay). For example, the formulations of the present disclosure, after storage for a specified time (e.g., 1 to 36 months) at -80°C, -30°C, -20°C, 5°C, 25°C, 40°C, 45°C, etc., if the anti-EBOV antibody contained in the formulation retains 90%, 95% or more of the relative ADCC potency compared to the antibody before such storage, it may be considered stable.

[0129] Similarly, the relative efficacy of an antibody in a pseudovirus neutralization assay can be measured to evaluate stability. For example, the formulations of the present disclosure are considered stable if, after storage at -80°C, -30°C, -20°C, 5°C, 25°C, 40°C, 45°C, etc. for a defined period of time (e.g., 1 to 36 months), the anti-EBOV antibody contained in the formulation retains 90%, 95% or more relative pseudovirus neutralizing activity compared to the antibody prior to such storage.

[0130] Additional methods for evaluating the stability of an antibody in a formulation are shown in the examples presented below.

[0131] The liquid pharmaceutical formulations of the present disclosure exhibit low to moderate viscosities in certain embodiments. As used herein, "viscosity" can be "kinematic viscosity" or "absolute viscosity". "Kinematic viscosity" is a measure of the resistance to flow of a fluid under the influence of gravity. When two fluids of equal volume are placed in the same capillary viscometer and allowed to flow by gravity, the more viscous fluid takes longer to flow through the capillary than the less viscous fluid. For example, if one fluid takes 200 seconds to complete its flow and another fluid takes 400 seconds, the second fluid has twice the viscosity of the first fluid on the kinematic viscosity scale. "Absolute viscosity", sometimes also called dynamic viscosity or simple viscosity, is the product of the kinematic viscosity and the fluid density (absolute viscosity = kinematic viscosity × density). The magnitude of kinematic viscosity is L 2 / T, where L is length and T is time. Generally, kinematic viscosity is expressed in centistokes (cSt). The SI unit of kinematic viscosity is mm 2 / s, which is 1 cSt. Absolute viscosity is expressed in units of centipoise (cP). The SI unit of absolute viscosity is millipascal-second (mPa-s), and 1 cP = 1 mPa-s.

[0132] As used herein, in connection with the fluid formulations of the present disclosure, a low level of viscosity exhibits an absolute viscosity of less than about 20 centipoise (cP). For example, the fluid formulations disclosed herein are considered to have a "low viscosity" when the formulation exhibits an absolute viscosity of about 20 cP, about 19 cP, about 18 cP, about 15 cP, about 12 cP, about 10 cP, about 9 cP, about 8 cP, or less, as measured using standard viscosity measurement techniques. As used herein, in connection with the fluid formulations of the present disclosure, a medium level of viscosity exhibits an absolute viscosity of from about 35 cP to about 20 cP. For example, the fluid formulations disclosed herein are considered to have a "medium viscosity" when the formulation exhibits an absolute viscosity of about 34 cP, about 33 cP, about 32 cP, about 31 cP, about 30 cP, about 29 cP, about 28 cP, about 27 cP, about 26 cP, about 25 cP, about 24 cP, about 23 cP, about 22 cP, about 21 cP, about 20 cP, about 19 cP, 18 cP, about 17 cP, about 16 cP, or about 15 cP, as measured using standard viscosity measurement techniques. The formulations provided herein can have a low viscosity, such as a viscosity of about 2 cP, for example.

[0133] Exemplary formulations According to one aspect of the present disclosure, a pharmaceutical formulation is a stable, low-viscosity, generally physiologically isotonic liquid formulation comprising (i) a human antibody that specifically binds to EBOV (e.g., H1H17203P, H1H17139P, and / or H1H17161P) at a concentration of up to 250 mg / mL ± 45 mg / mL, (ii) a histidine buffer system that provides sufficient buffering at about pH 6.0 ± 0.3, (iii) an organic co-solvent that protects the structural integrity of the antibody, and (iv) a sugar as a stabilizer.

[0134] According to one embodiment, a stable liquid pharmaceutical formulation has a pH of 6.0 ± 0.3 and contains (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 50 mg / mL ± 7.5 mg / mL total anti-EBOV antibody, wherein the antibody is a human IgG1 antibody comprising (i) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 2 / 10. In one embodiment, the anti-EBOV antibody comprises HCDR1 of SEQ ID NO: 4, HCDR2 of SEQ ID NO: 6, HCDR3 of SEQ ID NO: 8, LCDR1 of SEQ ID NO: 12, LCDR2 of SEQ ID NO: 14, and LCDR3 of SEQ ID NO: 16.

[0135] According to one embodiment, a stable liquid pharmaceutical formulation has a pH of 6.0 ± 0.3 and contains (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 50 mg / mL ± 7.5 mg / mL total anti-EBOV antibody, wherein the antibody is a human IgG1 antibody comprising (ii) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 20 / 28. In one embodiment, the anti-EBOV antibody comprises HCDR1 of SEQ ID NO: 22, HCDR2 of SEQ ID NO: 24, HCDR3 of SEQ ID NO: 26, LCDR1 of SEQ ID NO: 30, LCDR2 of SEQ ID NO: 32, and LCDR3 of SEQ ID NO: 34.

[0136] According to one embodiment, a stable liquid pharmaceutical formulation has a pH of 6.0 ± 0.3 and contains (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 50 mg / mL ± 7.5 mg / mL total anti-EBOV antibody, wherein the antibody is a human IgG1 antibody comprising (iii) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 38 / 46. In one embodiment, the anti-EBOV antibody comprises HCDR1 of SEQ ID NO: 40, HCDR2 of SEQ ID NO: 42, HCDR3 of SEQ ID NO: 44, LCDR1 of SEQ ID NO: 48, LCDR2 of SEQ ID NO: 50, and LCDR3 of SEQ ID NO: 52.

[0137] According to one embodiment, a stable liquid pharmaceutical formulation has a pH of 6.0 ± 0.3 and contains (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 50 mg / mL ± 7.5 mg / mL total anti-EBOV antibody, wherein the first antibody is a human IgG1 antibody comprising (i) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 2 / 10, and the second antibody is a human IgG1 antibody comprising (ii) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 20 / 28. In one embodiment, the first anti-EBOV antibody comprises HCDR1 of SEQ ID NO: 4, HCDR2 of SEQ ID NO: 6, HCDR3 of SEQ ID NO: 8, LCDR1 of SEQ ID NO: 12, LCDR2 of SEQ ID NO: 14, and LCDR3 of SEQ ID NO: 16, and the second anti-EBOV antibody comprises HCDR1 of SEQ ID NO: 22, HCDR2 of SEQ ID NO: 24, HCDR3 of SEQ ID NO: 26, LCDR1 of SEQ ID NO: 30, LCDR2 of SEQ ID NO: 32, and LCDR3 of SEQ ID NO: 34.

[0138] According to one embodiment, a stable liquid pharmaceutical formulation has a pH of 6.0 ± 0.3 and contains (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 50 mg / mL ± 7.5 mg / mL total anti-EBOV antibody, wherein the first antibody is a human IgG1 antibody comprising (i) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 2 / 10, and the second antibody is a human IgG1 antibody comprising (iii) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 38 / 46. In one embodiment, the first anti-EBOV antibody comprises HCDR1 of SEQ ID NO: 4, HCDR2 of SEQ ID NO: 6, HCDR3 of SEQ ID NO: 8, LCDR1 of SEQ ID NO: 12, LCDR2 of SEQ ID NO: 14, and LCDR3 of SEQ ID NO: 16, and the second anti-EBOV antibody comprises HCDR1 of SEQ ID NO: 40, HCDR2 of SEQ ID NO: 42, HCDR3 of SEQ ID NO: 44, LCDR1 of SEQ ID NO: 48, LCDR2 of SEQ ID NO: 50, and LCDR3 of SEQ ID NO: 52.

[0139] According to one embodiment, a stable liquid pharmaceutical formulation has a pH of 6.0 ± 0.3 and contains (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 50 mg / mL ± 7.5 mg / mL total anti-EBOV antibodies, wherein the first antibody is a human IgG1 antibody comprising (ii) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 20 / 28, and the second antibody is a human IgG1 antibody comprising (iii) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 38 / 46. In one embodiment, the first anti-EBOV antibody comprises HCDR1 of SEQ ID NO: 22, HCDR2 of SEQ ID NO: 24, HCDR3 of SEQ ID NO: 26, LCDR1 of SEQ ID NO: 30, LCDR2 of SEQ ID NO: 32, and LCDR3 of SEQ ID NO: 34, and the second anti-EBOV antibody comprises HCDR1 of SEQ ID NO: 40, HCDR2 of SEQ ID NO: 42, HCDR3 of SEQ ID NO: 44, LCDR1 of SEQ ID NO: 48, LCDR2 of SEQ ID NO: 50, and LCDR3 of SEQ ID NO: 52.

[0140] According to one embodiment, a stable liquid pharmaceutical formulation is provided that contains, at pH 6.0 ± 0.3, (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 50 mg / mL ± 7.5 mg / mL total anti-EBOV antibodies, where the first antibody is a human IgG1 antibody comprising (i) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 2 / 10, the second antibody is a human IgG1 antibody comprising (ii) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 20 / 28, and the third antibody is a human IgG1 antibody comprising (iii) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 38 / 46. In one embodiment, the first anti-EBOV antibody comprises HCDR1 of SEQ ID NO: 4, HCDR2 of SEQ ID NO: 6, HCDR3 of SEQ ID NO: 8, LCDR1 of SEQ ID NO: 12, LCDR2 of SEQ ID NO: 14, and LCDR3 of SEQ ID NO: 16, the second anti-EBOV antibody comprises HCDR1 of SEQ ID NO: 22, HCDR2 of SEQ ID NO: 24, HCDR3 of SEQ ID NO: 26, LCDR1 of SEQ ID NO: 30, LCDR2 of SEQ ID NO: 32, and LCDR3 of SEQ ID NO: 34, and the third anti-EBOV antibody comprises HCDR1 of SEQ ID NO: 40, HCDR2 of SEQ ID NO: 42, HCDR3 of SEQ ID NO: 44, LCDR1 of SEQ ID NO: 48, LCDR2 of SEQ ID NO: 50, and LCDR3 of SEQ ID NO: 52.

[0141] According to one embodiment, a stable liquid pharmaceutical formulation has a pH of 6.0 ± 0.3 and contains (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 100 mg / mL ± 15 mg / mL total anti-EBOV antibody, where the antibody is a human IgG1 antibody comprising (i) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 2 / 10. In one embodiment, the anti-EBOV antibody comprises HCDR1 of SEQ ID NO: 4, HCDR2 of SEQ ID NO: 6, HCDR3 of SEQ ID NO: 8, LCDR1 of SEQ ID NO: 12, LCDR2 of SEQ ID NO: 14, and LCDR3 of SEQ ID NO: 16.

[0142] According to one embodiment, a stable liquid pharmaceutical formulation has a pH of 6.0 ± 0.3 and contains (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 100 mg / mL ± 15 mg / mL total anti-EBOV antibody, where the antibody is a human IgG1 antibody comprising (ii) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 20 / 28. In one embodiment, the anti-EBOV antibody comprises HCDR1 of SEQ ID NO: 22, HCDR2 of SEQ ID NO: 24, HCDR3 of SEQ ID NO: 26, LCDR1 of SEQ ID NO: 30, LCDR2 of SEQ ID NO: 32, and LCDR3 of SEQ ID NO: 34.

[0143] According to one embodiment, a stable liquid pharmaceutical formulation has a pH of 6.0 ± 0.3 and contains (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 100 mg / mL ± 15 mg / mL total anti-EBOV antibody, wherein the antibody is a human IgG1 antibody comprising (iii) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 38 / 46. In one embodiment, the anti-EBOV antibody comprises HCDR1 of SEQ ID NO: 40, HCDR2 of SEQ ID NO: 42, HCDR3 of SEQ ID NO: 44, LCDR1 of SEQ ID NO: 48, LCDR2 of SEQ ID NO: 50, and LCDR3 of SEQ ID NO: 52.

[0144] According to one embodiment, a stable liquid pharmaceutical formulation has a pH of 6.0 ± 0.3 and contains (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 100 mg / mL ± 15 mg / mL total anti-EBOV antibody, wherein the first antibody is a human IgG1 antibody comprising (i) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 2 / 10, and the second antibody is a human IgG1 antibody comprising (ii) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 20 / 28. In one embodiment, the first anti-EBOV antibody comprises HCDR1 of SEQ ID NO: 4, HCDR2 of SEQ ID NO: 6, HCDR3 of SEQ ID NO: 8, LCDR1 of SEQ ID NO: 12, LCDR2 of SEQ ID NO: 14, and LCDR3 of SEQ ID NO: 16, and the second anti-EBOV antibody comprises HCDR1 of SEQ ID NO: 22, HCDR2 of SEQ ID NO: 24, HCDR3 of SEQ ID NO: 26, LCDR1 of SEQ ID NO: 30, LCDR2 of SEQ ID NO: 32, and LCDR3 of SEQ ID NO: 34.

[0145] According to one embodiment, a stable liquid pharmaceutical formulation has a pH of 6.0 ± 0.3 and contains (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 100 mg / mL ± 15 mg / mL total anti-EBOV antibody, where the first antibody is a human IgG1 antibody comprising (i) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 2 / 10, and the second antibody is a human IgG1 antibody comprising (iii) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 38 / 46. In one embodiment, the first anti-EBOV antibody comprises HCDR1 of SEQ ID NO: 4, HCDR2 of SEQ ID NO: 6, HCDR3 of SEQ ID NO: 8, LCDR1 of SEQ ID NO: 12, LCDR2 of SEQ ID NO: 14, and LCDR3 of SEQ ID NO: 16, and the second anti-EBOV antibody comprises HCDR1 of SEQ ID NO: 40, HCDR2 of SEQ ID NO: 42, HCDR3 of SEQ ID NO: 44, LCDR1 of SEQ ID NO: 48, LCDR2 of SEQ ID NO: 50, and LCDR3 of SEQ ID NO: 52.

[0146] According to one embodiment, a stable liquid pharmaceutical formulation has a pH of 6.0 ± 0.3 and contains (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 100 mg / mL ± 15 mg / mL total anti-EBOV antibody, where the first antibody is a human IgG1 antibody comprising (ii) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 20 / 28, and the second antibody is a human IgG1 antibody comprising (iii) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 38 / 46. In one embodiment, the first anti-EBOV antibody comprises HCDR1 of SEQ ID NO: 22, HCDR2 of SEQ ID NO: 24, HCDR3 of SEQ ID NO: 26, LCDR1 of SEQ ID NO: 30, LCDR2 of SEQ ID NO: 32, and LCDR3 of SEQ ID NO: 34, and the second anti-EBOV antibody comprises HCDR1 of SEQ ID NO: 40, HCDR2 of SEQ ID NO: 42, HCDR3 of SEQ ID NO: 44, LCDR1 of SEQ ID NO: 48, LCDR2 of SEQ ID NO: 50, and LCDR3 of SEQ ID NO: 52.

[0147] According to one embodiment, a stable liquid pharmaceutical formulation is provided that contains, at pH 6.0 ± 0.3, (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 100 mg / mL ± 15 mg / mL total anti-EBOV antibodies, where the first antibody is a human IgG1 antibody that includes (i) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 2 / 10, the second antibody is a human IgG1 antibody that includes (ii) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 20 / 28, and the third antibody is a human IgG1 antibody that includes (iii) three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCVR / LCVR amino acid sequence pair of SEQ ID NO: 38 / 46. In one embodiment, the first anti-EBOV antibody includes HCDR1 of SEQ ID NO: 4, HCDR2 of SEQ ID NO: 6, HCDR3 of SEQ ID NO: 8, LCDR1 of SEQ ID NO: 12, LCDR2 of SEQ ID NO: 14, and LCDR3 of SEQ ID NO: 16, the second anti-EBOV antibody includes HCDR1 of SEQ ID NO: 22, HCDR2 of SEQ ID NO: 24, HCDR3 of SEQ ID NO: 26, LCDR1 of SEQ ID NO: 30, LCDR2 of SEQ ID NO: 32, and LCDR3 of SEQ ID NO: 34, and the third anti-EBOV antibody includes HCDR1 of SEQ ID NO: 40, HCDR2 of SEQ ID NO: 42, HCDR3 of SEQ ID NO: 44, LCDR1 of SEQ ID NO: 48, LCDR2 of SEQ ID NO: 50, and LCDR3 of SEQ ID NO: 52.

[0148] Additional non-limiting examples of pharmaceutical formulations encompassed by the present disclosure are described elsewhere in this specification, including the examples presented below.

[0149] Containers and Administration Methods The pharmaceutical formulations of the present disclosure may be contained within any container suitable for the storage of pharmaceuticals and other therapeutic compositions. For example, the pharmaceutical formulations may be contained within a sealed and sterilized plastic or glass container having a predetermined volume, such as a vial, ampoule, syringe, cartridge, or bottle. Different types of vials, such as transparent or opaque (e.g., amber-colored) glass or plastic vials, may be used to contain the formulations of the present disclosure. Similarly, any type of syringe may be used to contain or administer the pharmaceutical formulations of the present disclosure.

[0150] The pharmaceutical formulations of the present disclosure may be contained within a "normal tungsten" syringe, or a "low tungsten" syringe. As will be appreciated by those skilled in the art, the process of manufacturing a glass syringe generally involves the use of a hot tungsten rod to pierce the glass, through which liquid can be drawn and expelled from the syringe. This process results in the deposition of trace amounts of tungsten on the inner surface of the syringe. Subsequent washing and other processing steps can be used to reduce the amount of tungsten in the syringe. As used herein, the term "normal tungsten" means that the syringe contains 500 parts per billion (ppb) or more of tungsten. The term "low tungsten" means that the syringe contains less than 500 ppb of tungsten. For example, according to the present disclosure, a low tungsten syringe may contain less than about 490, 480, 470, 460, 450, 440, 430, 420, 410, 390, 350, 300, 250, 200, 150, 100, 90, 80, 70, 60, 50, 40, 30, 20, 10 ppb, or less, of tungsten.

[0151] The rubber plunger used in a syringe and the rubber stopper used to close the opening of a vial may be coated to prevent contamination of the pharmaceutical contents of the syringe or vial, or may be coated to maintain their stability. Thus, according to certain embodiments, the pharmaceutical formulations of the present disclosure may be contained within a syringe containing a coated plunger or within a vial sealed with a coated rubber stopper. For example, the plunger or stopper may be coated with a fluorocarbon film. Examples of coated stoppers or plungers suitable for use in vials and syringes containing the pharmaceutical formulations of the present disclosure are referred to, for example, in U.S. Pat. Nos. 4,997,423, 5,908,686, 6,286,699, 6,645,635, and 7,226,554, the contents of which are hereby incorporated by reference in their entirety. Certain exemplary coated rubber stoppers and plungers that may be used in the context of the present disclosure are commercially available under the trade name "FluroTec®" from West Pharmaceutical Services, Inc. (Exton, PA). FluroTec® is an example of a fluorocarbon coating used to minimize or prevent the adhesion of pharmaceuticals to rubber surfaces.

[0152] According to certain embodiments of the present disclosure, the pharmaceutical formulation may be contained within a low tungsten syringe containing a fluorocarbon-coated plunger.

[0153] The pharmaceutical preparation can be administered to a patient via parenteral routes such as injection (e.g., subcutaneous, intravenous, intramuscular, intraperitoneal, etc.), or via transdermal, mucosal, nasal, pulmonary, or oral administration. The pharmaceutical preparation of the present disclosure can be delivered subcutaneously using a number of reusable pen-type delivery devices or autoinjector devices. Examples include, but are not limited to, AUTOPEN™ (Owen Mumford, Inc., Woodstock, UK), DISETRONIC™ pen (Disetronic Medical Systems, Burgdorf, Switzerland), HUMALOG MIX 75 / 25™ pen, HUMALOG™ pen, HUMALIN 70 / 30™ pen (Eli Lilly and Co., Indianapolis, Indiana), NOVOPEN™ I, II, and III (Novo Nordisk, Copenhagen, Denmark), NOVOPEN JUNIOR™ (Novo Nordisk, Copenhagen, Denmark), BD™ pen (Becton Dickinson, Franklin Lakes, New Jersey), OPTIPEN™, OPTIPEN PRO™, OPTIPEN STARLET™, and OPTICLIK™ (sanofi-aventis, Frankfurt, Germany). Examples of disposable pen delivery devices or autoinjector devices having use in subcutaneous delivery of the pharmaceutical composition of the present disclosure include, but are not limited to, SOLOSTAR™ pen (sanofi-aventis), FLEXPEN™ (Novo Nordisk), and KWIKPEN™ (Eli Lilly), SURECLICK™ Autoinjector (Amgen, Thousand Oaks, California), PENLET™ (Haselmeier, Stuttgart, Germany), EPIPEN (Dey, L.P.) and HUMIRA™ Pen (Abbott Labs, Abbott Park, Illinois).

[0154] The use of a microinjector for delivering the pharmaceutical formulations of the present disclosure is also contemplated herein. As used herein, the term "microinjector" means a subcutaneous delivery device designed to slowly administer a large volume (e.g., up to about 2.5 mL or more) of a therapeutic formulation over an extended period (e.g., about 10, 15, 20, 25, 30 minutes or more). See, e.g., U.S. 6,629,949, US 6,659,982, and Meehan et al., J. Controlled Release 46:107-116 (1996). Microinjectors are particularly useful for delivering multi-dose therapeutic proteins contained within highly concentrated solutions (e.g., about 100, 125, 150, 175, 200 mg / mL or more) or viscous solutions.

[0155] In certain embodiments, a stable liquid pharmaceutical formulation of any of the foregoing aspects is contained in a sterile glass vial and administered as an IV infusion. Exemplary dosages include 30,000 mg, 25,000 mg, 20,000 mg, 15,000 mg, 13,500 mg, 12,500 mg, 10,000 mg, 7,500 mg, 5000 mg, 2500 mg, 1450 mg, 1000 mg, 725 mg, 600 mg, 500 mg, 250 mg, 200 mg, 150 mg, 100 mg, 75 mg, 50 mg, or 25 mg.

[0156] In one embodiment, the container is a 20 mL type 1 borosilicate glass vial. In certain embodiments, the container is a 2 mL, 5 mL, or 10 mL type 1 borosilicate glass vial comprising a chlorobutyl stopper with a FluroTec® coating.

[0157] In one embodiment, the liquid pharmaceutical formulation of the present disclosure containing anti-EBOV antibodies at about 25 mg / mL, 50 mg / mL, 100 mg / mL, or 150 mg / mL is administered intravenously and may be contained in a glass vial. In some embodiments, the present disclosure provides a glass vial containing a stable liquid formulation containing, at a pH of about 6.0, 50 mg / mL, 100 mg / mL, or 150 mg / mL total anti-EBOV antibodies, 10 mM histidine, 10% sucrose, and 0.1% polysorbate 80.

[0158] In some embodiments, each antibody is administered at 50 mg / kg body weight. In one embodiment, two antibodies are co-formulated such that the final formulation is administered at 50 mg / kg body weight for each antibody. Thus, the final dose administered to the patient is 100 mg / kg body weight, and the two antibodies in the formulation are in a 1:1 ratio. In one embodiment, the co-formulated antibodies are delivered intravenously over about 2 hours.

[0159] In some embodiments, three antibodies are co-formulated such that the final formulation is administered at 50 mg / kg body weight for each antibody. Thus, the final dose administered to the patient is 150 mg / kg body weight, and the three antibodies in the formulation are in a 1:1:1 ratio. In one embodiment, the co-formulated antibodies are delivered intravenously over about 2 hours.

[0160] In some aspects, a patient weighing about 90 kg receiving a dose of 150 mg / kg receives an administration of about 13,500 mg. In some aspects, a patient weighing about 45 kg receiving 150 mg / kg receives an administration of about 6,750 mg. In some aspects, a patient may receive up to 30,000 mg of the administration.

[0161] In certain embodiments, the three antibodies are prepared in glass vials. In certain embodiments, each vial may contain 725 mg of total antibody, i.e., the three antibodies in a 1:1:1 ratio, in a volume of 14.5 mL, and the final antibody concentration is 50 mg / mL. This may be administered intravenously to a patient over 2 hours.

[0162] In certain embodiments, the three antibodies are prepared in glass vials. In certain embodiments, each vial may contain 1450 mg of total antibody, i.e., the three antibodies in a 1:1:1 ratio, in a volume of 14.5 mL, and the final antibody concentration is 100 mg / mL. This may be administered intravenously to a patient over 2 hours.

[0163] In certain embodiments, the present disclosure provides an autoinjector comprising any of the liquid formulations described herein. In some embodiments, the present disclosure provides an autoinjector containing a stable liquid formulation having a pH of about 6.0 and containing a total anti-EBOV antibody of about 50 mg / mL, about 100 mg / mL, about 150 mg / mL, or about 175 mg / mL, about 10 mM histidine, about 10% sucrose, and about 0.1% polysorbate 80.

[0164] In certain embodiments, the present disclosure provides an autoinjector comprising any of the liquid formulations described herein. In some embodiments, the present disclosure provides an autoinjector containing a stable liquid formulation having a pH of about 6.0 and containing a total anti-EBOV antibody of 50 mg / mL or 100 mg / mL, 10 mM histidine, 10% sucrose, and 0.1% polysorbate 80.

[0165] In certain embodiments, the present disclosure provides a prefilled syringe containing any of the liquid formulations described herein. In some embodiments, the present disclosure provides a prefilled syringe containing a stable liquid formulation having an anti-EBOV antibody at a pH of about 6.0, at about 50 mg / mL, about 100 mg / mL, about 150 mg / mL, or about 175 mg / mL, about 10 mM histidine, about 10% sucrose, and about 0.1% polysorbate 80. In certain embodiments, the syringe is a 1 mL or 2.25 mL long glass syringe filled with a 27-gauge thin-walled needle, a fluorocarbon-coated rubber plunger, and a rubber needle shield.

[0166] In one embodiment, a liquid pharmaceutical formulation containing a total anti-EBOV antibody of about 100 mg / mL ± 15 mg / mL is administered in a prefilled syringe at a volume of up to approximately 2 mL. In certain embodiments, the syringe is a 1 mL or 2.25 mL long glass syringe filled with a 27-gauge thin-walled needle, a fluorocarbon-coated rubber plunger, and a rubber needle shield. In one embodiment, the syringe is an OMPI 1 mL long glass syringe filled with a 27-gauge needle, an FM27 rubber needle shield, and a 4023 / 50 rubber plunger coated with FLUROTEC®.

[0167] In one embodiment, a liquid pharmaceutical formulation containing an anti-EBOV antibody of about 50 mg / mL ± 7.5 mg / mL is administered in a prefilled syringe at a volume of up to approximately 2 mL. In one embodiment, the syringe is a 1 mL or 2.25 mL long glass syringe filled with a 27-gauge thin-walled needle, a fluorocarbon-coated rubber plunger, and a rubber needle shield. In one embodiment, the syringe is an OMPI 1 mL long glass syringe filled with a 27-gauge needle, an FM27 rubber needle shield, and a 4023 / 50 rubber plunger coated with FLUROTEC®.

[0168] Compatibility In one embodiment, a stable formulation is a liquid solution containing 50 mg / mL of total anti-EBOV antibody (e.g., 16.7 mg / mL each of H1H17203P, H1H17139P, and / or H1H17161P, or approximately 25 mg / mL each of any two of H1H17203P, H1H17139P, and H1H17161P) for IV administration. In some embodiments, the IV admixture solution has compatibility (including stability during use) with diluents and materials used in administration systems (IV bags, sets, and filters) including 0.9% sodium chloride, 5% dextrose or lactated Ringer's solution, PVC (polyvinyl chloride), and polyethylene.

[0169] Exemplary dosages include 10 mg / kg, 30 mg / kg, 50 mg / kg, and 150 mg / kg. The stability during use of the anti-EBOV formulation during IV delivery supports administration of clinical dosages. In some examples, a 0.9% sodium chloride PVC IV bag containing the diluted antibody can first be held at 5 °C for 24 hours and then incubated at 25 °C for at least 8 hours. After these incubations, each of the drip sets can be connected to the IV bag, the diluted DP can be prepared, and held at room temperature for 1 hour. The diluted DP solution can then be pump infused through each drip set set at 25 mL / hr and 500 mL / hr. These drip sets contain basic materials (PVC with DEHP, PVC with TOTM, and polyethylene) including drip sets used for IV delivery of antibody combinations in clinical use. The drip set can include a 0.2 μm polyethersulfone in-line filter.

[0170] In some embodiments, a 50 mg / mL anti-EBOV formulation comprising at least one anti-EBOV antibody diluted with 0.9% sodium chloride injection to either a concentration of 2.2 mg / mL or 23.7 mg / mL is physically and chemically stable when verified under these conditions within the proposed dosage range and administration conditions. The anti-EBOV formulation, in some embodiments, shows no significant change in quality characteristics after dilution in an IV bag, storage in an IV bag at 2 - 8 °C for 24 hours and at 25 °C for at least 8 hours, holding in a drip set for 1 hour, or delivery at a pump rate of 25 mL / hr to 500 mL / hr.

[0171] In some embodiments, additional flexibility is provided by using various diluents for the dosage preparation of co-formulated antibodies and to support potential temperature excursions (higher than 25 °C) that may occur during IV administration. When the anti-EBOV antibody was diluted at 40 °C with 0.9% sodium chloride, 5% dextrose, or lactated Ringer's, the formulation continued to show stability. IV bags containing the diluted antibody can first be held at 5 °C for 24 hours and then incubated at 40 °C for at least 6 hours. After incubation, each of the drip sets can be connected to the IV bag, the diluted antibody can be prepared, and held at room temperature for 1 hour. The diluted antibody solution can then be pump-injected through each drip set.

[0172] In some embodiments, a 50 mg / mL anti-EBOV antibody formulation comprising at least one anti-EBOV antibody diluted with 0.9% sodium chloride injection, 5% dextrose, or lactated Ringer's to either a concentration of 1.6 mg / mL or 27.9 mg / mL can be physically and chemically stable under the conditions tested within the proposed dosage range and administration conditions. The anti-EBOV formulation, in some embodiments, shows no significant change in quality characteristics after dilution in an IV bag, storage in an IV bag at 2 - 8 °C for 24 hours and at 40 °C for at least 6 hours, and being held in a drip set for 1 hour.

[0173] The IV compatibility characteristics (including stability during use) of the stable liquid pharmaceutical formulation support the following conclusions regarding dose preparation and IV administration (50 mg / mL). · IV bags made of PVC of 0.9% sodium chloride injection solution, 5% dextrose injection solution, and Ringer's lactate are compatible with IV administration of formulations containing at least one anti-EBOV antibody. · Anti-EBOV antibody formulations can be diluted in PVC IV bags containing IV diluents of 0.9% sodium chloride, 5% dextrose, or Ringer's lactate for IV administration to antibody concentrations as low as 2.2 mg / mL and as high as 23.7 mg / mL. · Diluted co-formulated antibodies prepared with 0.9% sodium chloride injection solution can be stored at room temperature up to 25°C for at least about 8 hours from preparation to the end of infusion, or at 2 - 8°C for at least about 24 hours from preparation to the end of infusion. · Diluted co-formulated antibodies prepared with 5% dextrose or Ringer's lactate can be stored at room temperature up to 25°C for at least about 4 hours from preparation to the end of infusion, or at 2 - 8°C for at least about 24 hours from preparation to the end of infusion. · Diluted co-formulated antibodies (2.2 mg / mL - 23.7 mg / mL) prepared with 0.9% sodium chloride, 5% dextrose, or Ringer's lactate solution are stable at 40°C for 6 hours. · Diluted co-formulated antibodies can be administered using an infusion set composed of any of PVC lined with DEHP-containing PVC, TOTM-containing PVC, or polyethylene. · Diluted co-formulated antibodies are compatible with the use of an in-line 0.2 μm polyethersulfone filter.

[0174] Therapeutic use of the pharmaceutical formulation The pharmaceutical formulations of the present disclosure are particularly useful for the treatment, prevention, or amelioration of EBOV or any symptoms associated with EBOV.

Examples

[0175] The following examples are presented to provide a complete disclosure and description to those skilled in the art of how to make and use the methods and compositions disclosed herein and are not intended to limit the scope of what the inventors regard as their invention. Efforts have been made to ensure accuracy with respect to the numbers used (e.g., amounts, temperatures, etc.), but some experimental errors and deviations should be accounted for. Unless otherwise indicated, parts are parts by mole, molecular weight is average molecular weight, temperature is in degrees Celsius, and pressure is at or near atmospheric pressure.

[0176] Example 1: Development of an Anti-EBOV Antibody Formulation The goal of the formulation activities was to develop a formulation having the following characteristics: · A liquid formulation that is stable even after being exposed to stresses such as temperature cycling, extreme temperatures, agitation during transportation, etc., where the therapeutic agent is exposed during transportation from the manufacturing facility to a remote field clinic where Ebola exposure occurs and / or where there are Ebola patients. · A liquid formulation having a concentration of anti-EBOV antibody sufficient to deliver doses of 25 mg to 30,000 mg, such as about 7500 mg, about 5000 mg, about 3000 mg, about 2000 mg, about 1500 mg, 1000 mg, about 800 mg, about 750 mg, about 500 mg, about 250 mg, about 200 mg, about 150 mg, about 100 mg, about 75 mg, about 50 mg, or about 25 mg by intravenous infusion. · A formulation with approximately equal osmotic pressure that is stable when diluted with diluents commonly used for intravenous infusion, such as 0.9% sodium chloride injection solution or 5% dextrose injection solution or lactated Ringer's injection solution. · A stable formulation that is compatible with type 1 clear glass vials as packaging and standard serum stoppers. And · A sterile formulation (DP) solution that supports long-term stability. o A formulation that minimizes the high molecular weight (HMW) species of the antibody when exposed to handling stress and heat stress. o A formulation that minimizes changes in the relative distribution of the charged species of the antibody when exposed to heat stress. And A formulation that maintains biological activity when subjected to rough transportation and heat stress in extreme environments.

[0177] Throughout formulation development, three important protein stress conditions (meaning extreme handling conditions to which an antibody formulation can be exposed during handling, manufacturing, transportation, storage, and labeling) were adopted to develop and qualify antibody formulations to assess the impact of potential real-world stresses on the stability of formulations used in remote regions of the world. These stress conditions include the following: · Stirring (vortexing) of the protein solution at room temperature. Vortexing in glass vials exceeds the stirring that occurs during protein handling and manufacturing. · Incubating the protein solution at elevated temperatures (37 °C, 40 °C, or 45 °C) compared to the proposed DP storage conditions (2 °C to 8 °C). · Exposing the protein to multiple freeze-thaw cycles. Since the protein undergoes at least one freeze-thaw cycle during DP manufacturing, multiple freeze-thaw cycles simulate and exceed the actual stress that the protein is expected to experience.

[0178] There were several major objectives in the initial formulation development work. 1. Selection of buffer and pH for each of the three anti-EBOV antibodies: Since the selection of buffer and pH can greatly affect protein stability, determining the optimal buffer type and pH is an important process. In these sections, experiments are presented that show the basis for the selection of the optimal buffer and pH for each antibody. 2. Selection of Surfactants or Organic Cosolvents for Each of the Three Anti-EBOV Antibodies: Surfactants such as polysorbate, or polysorbate, for example, are typically required to prevent precipitation or aggregation of proteins when stirred. Soluble proteins can be subject to stirring when handled, filtered, mixed, manufactured, transported, and administered. Antibody drug substances in simple buffer solutions can visibly become turbid with excessive stirring. Therefore, it was determined that it is important to stabilize each protein against handling and stirring. 3. Identification / Selection of Stabilizing / Isotonicity Excipients: The addition of sugars, salts, and amino acids was verified for their ability to improve the stability of each of the three antibodies against heat stress and to extend the shelf life of the drug product (DP). Experiments are presented herein to establish the rationale for the inclusion of these heat stabilizers and to identify the optimal concentrations in the final formulation. 4. Selection of Antibody Concentration: The effect of antibody concentration on the stability of formulations containing the selected excipients was verified. 5. Co-formulation of the Three Anti-EBOV Antibodies: The three anti-EBOV antibodies were co-formulated in a liquid formulation at two concentrations, selecting and combining buffers and pH, selecting surfactants and organic cosolvents, and identifying and selecting additional excipients and stabilizers to be tested.

[0179] Initial formulation development activities were conducted using each anti-EBOV antibody formulated individually at 100 mg / mL, screening for organic cosolvents, heat stabilizers, and buffers in the liquid formulations of each anti-EBOV antibody, identifying excipients that are compatible with the protein and maintain near-physiological osmotic pressure and low viscosity for intravenous and subcutaneous injection while enhancing its stability. Buffer conditions were also verified to determine the optimal pH for maximum protein stability (described in Example 6 herein).

[0180] Using the results of this initial formulation development effort, an initial formulation suitable for clinical trials was developed.

[0181] Knowledge obtained from the development of the initial formulation led to the identification of co-formulation of three antibodies at two concentrations, pH, surfactant concentration, and stabilizers for the later-stage formulation development activities, and excipients that enhance protein stability at both low and high protein concentrations and when exposed to stresses such as, for example, high temperature and agitation were identified (described in Examples 4-9).

[0182] Throughout formulation development, the formulations were evaluated for stress and storage stability. The methods used to evaluate stability in the formulation development studies are described in Example 3 of this specification. Examples 4-9 describe the storage and stress stability of the formulations.

[0183] Using the results obtained from these tests, a stable liquid formulation suitable for clinical use for intravenous (IV) administration was developed. Such formulations showed stability when exposed to heat or agitation stress.

[0184] Other properties of the formulations will be apparent from the description herein.

[0185] Anti-EBOV antibody: The anti-EBOV antibodies are described in US 2016 / 0215040, which is incorporated herein by reference in its entirety. Exemplary antibodies used in the following examples are the fully human anti-EBOV antibodies H1H17203P (REGN3470; having the HCVR amino acid sequence of SEQ ID NO: 2 and the LCVR amino acid sequence of SEQ ID NO: 10), H1H17139P (REGN3471; having the HCVR amino acid sequence of SEQ ID NO: 20 and the LCVR amino acid sequence of SEQ ID NO: 28), and H1H17161P (REGN3479; having the HCVR amino acid sequence of SEQ ID NO: 38 and the LCVR amino acid sequence of SEQ ID NO: 46), which include the sequences described in detail above.

[0186] Example 2: Exemplary Formulations In certain embodiments, the anti-EBOV antibody is formulated individually or co-formulated as an aqueous buffer formulation comprising, at pH 6.0 ± 0.3, (a) 5% ± 1% to 15% ± 3% w / v sucrose, (b) 5 mM ± 1 mM to 20 mM ± 4 mM histidine buffer, (c) 0.01% ± 0.005% to 0.5% ± 0.25% w / v polysorbate, and (d) 50 mg / mL ± 7.5 mg / mL total anti-EBOV antibody. In certain embodiments, the anti-EBOV antibody is formulated individually or co-formulated as an aqueous buffer formulation comprising, at pH 6.0 ± 0.3, (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 50 mg / mL ± 7.5 mg / mL total antibody. When co-formulated, the anti-EBOV antibodies are present in a 1:1:1 ratio.

[0187] In certain embodiments, the anti-EBOV antibody is formulated individually or co-formulated as an aqueous buffer formulation comprising, at pH 6.0 ± 0.3, (a) 5% ± 1% to 15% ± 3% w / v sucrose, (b) 5 mM ± 1 mM to 20 mM ± 4 mM histidine buffer, (c) 0.01% ± 0.005% to 0.5% ± 0.25% w / v polysorbate, and (d) 100 mg / mL ± 15 mg / mL total antibody. In certain embodiments, the anti-EBOV antibody is formulated individually or co-formulated as an aqueous buffer formulation comprising, at pH 6.0 ± 0.3, (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 100 mg / mL ± 15 mg / mL total antibody. When co-formulated, the anti-EBOV antibodies are present in a 1:1:1 ratio.

[0188] Exemplary formulations include the following: · A stable liquid pharmaceutical formulation comprising, at pH 6.0 ± 0.3, (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 50 mg / mL ± 7.5 mg / mL H1H17203P anti-EBOV antibody. · A stable liquid pharmaceutical formulation containing, at pH 6.0 ± 0.3, (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 50 mg / mL ± 7.5 mg / mL of H1H17139P anti-EBOV antibody. · A stable liquid pharmaceutical formulation containing, at pH 6.0 ± 0.3, (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 50 mg / mL ± 7.5 mg / mL of H1H17161P anti-EBOV antibody. · A stable liquid pharmaceutical formulation containing, at pH 6.0 ± 0.3, (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 50 mg / mL ± 7.5 mg / mL of the total anti-EBOV antibody of H1H17203P and H1H17139P. · A stable liquid pharmaceutical formulation containing, at pH 6.0 ± 0.3, (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 50 mg / mL ± 7.5 mg / mL of the total anti-EBOV antibody of H1H17203P and H1H17161P. · A stable liquid pharmaceutical formulation containing, at pH 6.0 ± 0.3, (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 50 mg / mL ± 7.5 mg / mL of the total anti-EBOV antibody of H1H17139P and H1H17161P. · A stable liquid pharmaceutical formulation containing, at pH 6.0 ± 0.3, (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 50 mg / mL ± 7.5 mg / mL of the total anti-EBOV antibody of H1H17203P, H1H17139P and H1H17161P. · A stable liquid pharmaceutical preparation containing (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 100 mg / mL ± 10 mg / mL of H1H17203P anti-EBOV antibody at pH 6.0 ± 0.3. · A stable liquid pharmaceutical preparation containing (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 100 mg / mL ± 15 mg / mL of H1H17139P anti-EBOV antibody at pH 6.0 ± 0.3. · A stable liquid pharmaceutical preparation containing (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 100 mg / mL ± 15 mg / mL of H1H17161P anti-EBOV antibody at pH 6.0 ± 0.3. · A stable liquid pharmaceutical preparation containing (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 100 mg / mL ± 15 mg / mL of the total anti-EBOV antibody of H1H17203P and H1H17139P at pH 6.0 ± 0.3. · A stable liquid pharmaceutical preparation containing (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 100 mg / mL ± 15 mg / mL of the total anti-EBOV antibody of H1H17203P and H1H17161P at pH 6.0 ± 0.3. · A stable liquid pharmaceutical preparation containing (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) 100 mg / mL ± 15 mg / mL of the total anti-EBOV antibody of H1H17139P and H1H17161P at pH 6.0 ± 0.3. A stable liquid pharmaceutical formulation containing, at pH 6.0 ± 0.3, (a) 10% ± 2% w / v sucrose, (b) 10 mM ± 2 mM histidine buffer, (c) 0.1% ± 0.05% w / v polysorbate, and (d) a total anti-EBOV antibody of 100 mg / mL ± 15 mg / mL of H1H17203P, H1H17139P, and H1H17161P.

[0189] Example 3: Methods used for evaluating formulation stability The following assays were applied to evaluate the stability of the formulation. · Color and appearance by visual inspection · pH · Increase in OD at 405 nm or turbidity measured by nephelometry · Particle matter analysis performed by microflow imaging (MFI) (reported as the number of particles obtained as is) and light obscuration method (HIAC) · Protein concentration by reverse phase - ultra high performance liquid chromatography (RP-UPLC) · Purity by size exclusion - ultra high performance liquid chromatography (SE-UPLC) or purity by reduced and non-reduced microchip capillary electrophoresis - sodium dodecyl sulfate (MCE-SDS) PAGE · Charge variant analysis by cation exchange chromatography - ultra high performance liquid chromatography (CEX-UPLC) or charge variant analysis by imaging capillary isoelectric focusing (iCIEF) · Potency by bioassay: The relative potency of each sample was determined using a bioassay and is defined as follows: (IC 50 Reference sample / IC 50 Sample) × 100%. The measured potency of the storage stability samples must be within 50% - 150% of the measured potency of the reference standard.

[0190] The physical stability of the formulation refers to characteristics such as color, appearance, pH, turbidity, and protein concentration. The presence of visible particles in the solution can be detected by visual inspection. The appearance passes the visual inspection if it is transparent to slightly milky white, essentially free of visible particles, and colorless to light yellow. Furthermore, turbidity measured by OD at 405 nm can also be used to detect particles in the solution. An increase in OD at 405 nm may indicate the presence of particles, an increase in opalescence, or discoloration of the test substance. MFI is used to measure subvisible particles with a size of 2 μm or larger. The anti-EBOV antibody protein concentration is measured by RP-UPLC assay and reported as the protein recovery rate compared to the starting material. In the RP-UPLC assay, the anti-EBOV antibody elutes from the RP column as a single peak. The protein concentration is determined from the total peak area of the antibody by comparing it to a calibration curve generated using an antibody standard. The recovery rate is calculated based on the measured protein concentration compared to the starting protein concentration.

[0191] Chemical stability refers to the formation of covalent modification forms of the protein (e.g., covalent aggregates, cleavage products, or charge variant forms) and non-covalent modification forms (e.g., non-covalent aggregates). High-molecular-weight and low-molecular-weight degradation products can be separated from the native antibody by SE-UPLC and MCE-SDS methods. The percentage of degraded anti-EBOV antibody in the SE-UPLC and MCE-SDS methods is calculated from the ratio of the area of all non-native peaks to the total area of all anti-EBOV antibody peaks. Charge variant forms of the anti-EBOV antibody are separated using CEX-UPLC and iCIEF. In the CEX-UPLC method, peaks with a retention time earlier than the main peak are labeled as "acidic" peaks, and peaks with a retention time later than the main peak are labeled as "basic" peaks. In the iCIEF method, peaks focused at a lower pI than the main peak are labeled as "acidic" peaks, and peaks focused at a higher pI than the main peak are labeled as "basic" peaks.

[0192] Example 4: Stability Test of Aqueous Formulation of Anti-EBOV Antibody All tests described in this section of this example refer to stability investigation tests conducted on H1H17203P formulation (DP: drug product), H1H17139P DP, and H1H17161P DP. Each DP was formulated and filled separately to provide stability data under real-time conditions, accelerated conditions, and stress stability conditions. The lots of drug substance (DS) used in these tests are representative of the DS manufactured for clinical use.

[0193] Formulation Development The H1H17203P antibody, H1H17139P antibody, and H1H17161P antibody are formulated for delivery by intravenous (IV) injection for the purpose of human first-in-human studies. The stability of the individually formulated and filled H1H17203P, H1H17139P, and H1H17161P formulations was evaluated in the clinical formulation. The results of analytical characterization indicated that this formulation provided appropriate stability for the H1H17203P antibody, H1H17139P antibody, and H1H17161P antibody under all validated conditions. The formulations of the H1H17203P antibody, H1H17139P antibody, and H1H17161P antibody contain 10 mM histidine, pH 6.0, 0.1% (w / v) polysorbate 80, and 10% (w / v) sucrose.

[0194] Stability Investigation All tests of FDS (formulated drug substance) and DP (formulation) outlined in this section refer to stability investigation tests conducted on the individually formulated and filled H1H17203P antibody, H1H17139P antibody, and H1H17161P antibody manufactured for development use. Formal stability tests to verify the H1H17203P antibody, H1H17139P antibody, and H1H17161P antibody manufactured for clinical use are considered below.

[0195] Stability tests were initiated to determine the storage stability, accelerated stability, and stress stability of individually formulated and filled investigational lots of 50 mg / mL H1H17203P DP, 50 mg / mL H1H17139P DP, and 50 mg / mL H1H17161P DP. The DS lots used in these tests are representative of the DS manufactured for clinical use. The DP was incubated under several elevated temperature conditions compared to storage temperature conditions. These accelerated conditions were selected to simulate conditions under which the DP would not be exposed beyond those during manufacture and handling, as well as to elucidate the degradation pathways of H1H17203P DP, H1H17139P DP, and H1H17161P DP. An overview of the storage conditions, accelerated conditions, and stress stability conditions for H1H17203P DP, H1H17139P DP, and H1H17161P DP is presented in Table 1, and the analytical plan is presented in Table 2.

[0196] (Table 1) Stability investigation tests for H1H17203P DP, H1H17139P DP, and H1H17161P DP TIFF2025090691000001.tif62167 a Frozen at -80 °C and thawed at room temperature.

[0197] (Table 2) Analytical plan for stability investigation tests for H1H17203P DP, H1H17139P DP, and H1H17161P DP TIFF2025090691000002.tif94167

[0198] Stability investigation for 50 mg / mL H1H17203P DP Storage stability investigation for 50 mg / mL H1H17203P DP Presents the stability investigation data for 50 mg / mL H1H17203P DP over 3 months. 50 mg / mL H1H17203P DP was physically and chemically stable when stored at 5 °C for 3 months (Table 3). No perceptible changes in physical or chemical stability were detected in any of the monitored characteristics.

[0199] Accelerated stability investigation for 50 mg / mL H1H17203P DP The analysis results of 50 mg / mL H1H17203P DP after incubation under accelerated conditions are presented in Table 6. After incubating H1H17203P DP at 45 °C for 28 days, increases of 2.1% and 1.6% were observed in the relative amounts of HMW and LMW species (SE-UPLC), and increases of 15.1% and 13.5% were observed in the proportions of acidic charge variant species as measured by CEX-UPLC and iCIEF, respectively. After incubating H1H17203P DP at 25 °C for 3 months, an increase of 0.3% was observed in the amounts of both HMW and LMW species (SE-UPLC), and increases of 2.9% and 3.0% were observed in the proportions of acidic charge variant species as measured by CEX-UPLC and iCIEF, respectively. From the results of the accelerated stability test, it was shown that the increase in the relative amounts of HMW and LMW species, as well as the increase in the formation of acidic charge variants, are the main degradation pathways of 50 mg / mL H1H17203P DP.

[0200] Stress stability investigation for 50 mg / mL H1H17203P DP The results of the stress stability investigation test are shown in Table 9. H1H17203P DP was physically and chemically stable when stirred for 120 minutes (vortex at atmospheric temperature). No perceivable changes in physical or chemical stability were detected in any of the monitored characteristics. H1H17203P DP was physically and chemically stable when exposed to 8 freeze / thaw cycles (freezing at -80°C and thawing at room temperature). No perceivable changes in physical or chemical stability were detected in any of the monitored characteristics.

[0201] Stability investigation of 50 mg / mL H1H17139P DP Storage stability investigation of 50 mg / mL H1H17139P DP Stability investigation data for 50 mg / mL H1H17139P DP over 3 months are shown. 50 mg / mL H1H17139P DP was physically and chemically stable when stored at 5°C for 3 months (Table 4). No perceivable changes in physical or chemical stability were detected in any of the monitored characteristics. 50 mg / mL H1H17139P DP maintained its potency over the 3-month evaluation period as determined by bioassay analysis.

[0202] Accelerated stability investigation of 50 mg / mL H1H17139P DP The analysis results of 50 mg / mL H1H17139P DP after incubation under accelerated conditions are presented in Table 7. After incubating H1H17139P DP at 45°C for 28 days, increases of 0.8% and 1.8% were observed in the relative amounts of HMW species and LMW species (SE-UPLC), respectively, and increases of 13.8% and 12.9% were observed in the proportions of acidic charge variant species as measured by CEX-UPLC and iCIEF, respectively. After incubating H1H17139P DP at 25°C for 3 months, increases of 0.4% and 0.5% were observed in the amounts of HMW species and LMW species (SE-UPLC), respectively, and increases of 2.6% and 3.1% were observed in the proportions of acidic charge variant species as measured by CEX-UPLC and iCIEF, respectively. H1H17139P maintained its potency as determined by bioassay analysis after incubation under each accelerated condition. From the results of the accelerated stability test, it was shown that the increases in the relative amounts of HMW species and LMW species, as well as the increase in the formation of acidic charge variants, were the main degradation pathways of 50 mg / mL H1H17139P DP.

[0203] Stress stability investigation for 50 mg / mL H1H17139P DP The results of the stress stability investigation test are shown in Table 10. H1H17139P DP was physically and chemically stable when stirred for 120 minutes (vortex at ambient temperature). No perceivable changes in physical stability or chemical stability were detected in any of the monitored characteristics. H1H17139P DP was physically and chemically stable when exposed to 8 freeze / thaw cycles (freezing at -80°C and thawing at room temperature). No perceivable changes in physical stability or chemical stability were detected in any of the monitored characteristics.

[0204] Stability investigation for 50 mg / mL H1H17161P DP Storage stability investigation for 50 mg / mL H1H17161P DP Shows the stability investigation data for 50 mg / mL H1H17161P DP over 3 months. 50 mg / mL H1H17161P DP was physically and chemically stable when stored at 5°C for 3 months (Table 5). No perceptible changes in physical or chemical stability were detected in any of the monitored characteristics. 50 mg / mL H1H17161P DP maintained its potency over the 3-month evaluation period as determined by bioassay analysis.

[0205] Accelerated stability investigation for 50 mg / mL H1H17161P DP The analysis results of 50 mg / mL H1H17161P DP after incubation under accelerated conditions are presented in Table 8. After incubating H1H17161P DP at 45°C for 28 days, increases of 1.0% and 1.8% were observed in the relative amounts of HMW and LMW species (SE-UPLC), and increases of 15.6% and 13.2% were observed in the proportions of acidic charge variant species as measured by CEX-UPLC and iCIEF, respectively. After incubating H1H17161P DP at 25°C for 3 months, increases of 0.7% and 0.5% were observed in the amounts of HMW and LMW species (SE-UPLC), and increases of 4.2% and 3.4% were observed in the proportions of acidic charge variant species as measured by CEX-UPLC and iCIEF, respectively. H1H17161P maintained its potency as determined by bioassay analysis after incubation under each accelerated condition. From the results of the accelerated stability test, it was shown that the increase in the relative amounts of HMW and LMW species, as well as the increase in the formation of acidic charge variants, were the main degradation pathways of 50 mg / mL H1H17161P DP.

[0206] Stress stability investigation for 50 mg / mL H1H17161P DP The results of the stress stability investigation test are shown in Table 11. H1H17161P DP was physically and chemically stable when stirred for 120 minutes (vortex at ambient temperature). No perceivable changes in physical or chemical stability were detected in any of the monitored characteristics. H1H17161P DP was physically and chemically stable when exposed to 8 freeze / thaw cycles (freezing at -80°C and thawing at room temperature). No perceivable changes in physical or chemical stability were detected in any of the monitored characteristics.

[0207] Conclusions of the stability investigation for H1H17203P DP, H1H17139P DP, and H1H17161P DP From the test results of the storage stability, accelerated stability, and stress stability of the DP, it is shown that the formulations of H1H17203P DP, H1H17139P DP, and H1H17161P DP can withstand limited exposure to room temperature without compromising physical or chemical stability. Furthermore, from the formulation tests of H1H17203P, H1H17139P, and H1H17161P, it is shown that H1H17203P DP, H1H17139P DP, and H1H17161P DP are stable when stored at 5°C for at least 3 months. H1H17203P DP, H1H17139P DP, and H1H17161P DP should be stored at 2°C - 8°C, and temperature exposure above 8°C should be limited.

[0208] (Table 3) Stability investigation of 50 mg / mL H1H17203P formulation stored at 5°C TIFF2025090691000003.tif178158

[0209] (Table 4) Stability investigation of 50 mg / mL H1H17139P formulation stored at 5°C TIFF2025090691000004.tif173159

[0210] (Table 5) Stability investigation of 50 mg / mL H1H17161P formulation stored at 5°C TIFF2025090691000005.tif181160

[0211] (Table 6) Stability investigation of 50 mg / mL H1H17203P formulation - Influence of accelerated conditions TIFF2025090691000006.tif165162

[0212] (Table 7) Stability investigation of 50 mg / mL H1H17139P formulation - Influence of accelerated conditions TIFF2025090691000007.tif162162

[0213] (Table 8) Stability investigation of 50 mg / mL H1H17161P formulation - Influence of accelerated conditions TIFF2025090691000008.tif172162

[0214] (Table 9) Stability investigation of 50 mg / mL H1H17203P formulation - Influence of stress conditions TIFF2025090691000009.tif160162

[0215] (Table 10) Stability investigation of 50 mg / mL H1H17139P formulation - Influence of stress conditions TIFF2025090691000010.tif159161

[0216] (Table 11) Stability investigation of 50 mg / mL H1H17161P formulation - Influence of stress conditions TIFF2025090691000011.tif164161

[0217] Conclusion The H1H17203P antibody, H1H17139P antibody, and H1H17161P antibody are manufactured as a liquid DP for IV administration. The H1H17203P DP contains 50 mg / mL H1H17203P antibody formulated in a solution containing 10 mM histidine, pH 6.0, 0.1% (w / v) polysorbate 80, and 10% (w / v) sucrose. The H1H17139P DP contains 50 mg / mL H1H17139P antibody formulated in a solution containing 10 mM histidine, pH 6.0, 0.1% (w / v) polysorbate 80, and 10% (w / v) sucrose. The H1H17161P DP contains 50 mg / mL H1H17161P antibody formulated in a solution containing 10 mM histidine, pH 6.0, 0.1% (w / v) polysorbate 80, and 10% (w / v) sucrose.

[0218] Based on the test results of this example, 50 mg / mL H1H17203P DP, 50 mg / mL H1H17139P DP, and 50 mg / mL H1H17161P DP are stable when stored at 2 - 8 °C for at least 12 months. Furthermore, the major degradation pathways identified under accelerated conditions were the formation of HMW and LMW species, as well as acidic charge variants.

[0219] Example 5: Stability Testing of an Aqueous Formulation Containing a Combination of 50 mg / mL Anti - EBOV Antibodies Three anti-EBOV antibodies, H1H17203P, H1H17139P, and H1H17161P, were formulated using 50 mg / mL total protein (16.7 mg / mL H1H17203P, 16.7 mg / mL H1H17139P, and 16.7 mg / mL H1H17161P), 10 mM histidine, pH 6.0, 0.1% (w / v) polysorbate 80, and 10% (w / v) sucrose. The three anti-EBOV monoclonal antibodies were formulated and mixed in one glass vial. The methods used to evaluate stability were developed to provide potential information regarding the antibodies of each component. However, many of the analytical methods cannot provide information regarding each individual antibody. When the results cannot be provided individually for each antibody, the analytical methods provide stability data regarding the entire formulation.

[0220] The physical stability of the formulation refers to characteristics such as color, appearance, pH, turbidity, and protein concentration. The presence of visible microparticles in the solution can be detected by visual inspection. The appearance passes the visual inspection if it is clear to slightly milky white, essentially free of visible particles, and colorless to pale yellow. Turbidity measured by an increase in OD at 405 nm can also be used to detect particles in the solution. An increase in OD at 405 nm may indicate the presence of microparticles, an increase in opalescence, or discoloration of the test substance. MFI is used to measure subvisible particles with a size of 2 μm or greater. The total protein concentration is measured by RP-UPLC assay and reported as the protein recovery rate compared to the starting material.

[0221] In the RP-UPLC assay, after elution from the reverse-phase column, the peaks of H1H17203P, H1H17139P, and H1H17161P cannot be separated from each other (Figure 1). The total protein concentration (H1H17203P, H1H17139P, and H1H17161P) is determined by comparing the peak areas with a calibration curve generated using the H1H17203P standard. Since the extinction coefficients of H1H17203P, H1H17139P, and H1H17161P are 1.50, 1.57, and 1.36, respectively, the extinction coefficient of the co-formulated H1H17203P, H1H17139P, and H1H17161P (1:1:1) would be approximately 1.48 (the average of the three extinction coefficients). Therefore, the H1H17203P standard (extinction coefficient 1.50) was selected to generate a standard curve to determine the total protein concentration in the co-formulated preparation. The recovery rate is calculated based on the measured total protein concentration compared to the starting concentration.

[0222] Chemical stability refers to the formation of covalent modification forms of proteins (e.g., covalent aggregates, cleavage products, or charge variant forms) and non-covalent modification forms (e.g., non-covalent aggregates). High molecular weight and low molecular weight degradation products can be separated from the native molecular weight products using the SE-UPLC method and the MCE-SDS method. The three formulation antibody preparations are characterized by SE-UPLC for total purity (native H1H17203P, H1H17139P, and H1H17161P) (i.e., the molecular weight purity of H1H17203P, H1H17139P, and H1H17161P is not determined individually). The reason is that the native species of H1H17203P, H1H17139P, and H1H17161P cannot be separated from each other (Figure 2). Similarly, the three formulation antibody preparations are characterized for total high molecular weight (HMW) species (H1H17203P HMW, H1H17139P HMW, and H1H17161P HMW) and total low molecular weight (LMW) species (H1H17203P LMW, H1H17139P LMW, and H1H17161P LMW). The reason is that the HMW or LMW species of H1H17203P, H1H17139P, and H1H17161P cannot be separated from each other (Figure 2). The ratio of total HMW species or total LMW species in the three formulation preparations is determined using the SE-UPLC method and calculated from the ratio of the area of the total HMW species or total LMW species to the total area of all peaks of H1H17203P, H1H17139P, and H1H17161P. The total purity determined by non-reducing MCE-SDS is calculated from the ratio of the intensity of the major bands of H1H17203P, H1H17139P, and H1H17161P to the total intensity of all bands. The total purity determined by reducing MCE-SDS is calculated from the ratio of the sum of the intensities of the heavy chain and light chain bands of H1H17203P, H1H17139P, and H1H17161P to the total intensity of all bands.

[0223] The iCIEF method did not have sufficient resolution to separate all charge variant forms of all three antibodies. Therefore, this analytical method was not used to evaluate the changes in the charge variant profile of the 3-formulation preparation sample. The charge variant forms of co-formulated H1H17203P, H1H17139P, and H1H17161P could be separated using the CEX-UPLC method. For H1H17203P, H1H17139P, and H1H17161P, peaks with a retention time earlier than the main peak were labeled as "acidic" peaks, and peaks with a retention time later than the main peak were labeled as "basic" peaks. The ratios of the acidic, main, and basic peaks were calculated by comparing the individual peak areas with the total peak area of each antibody.

[0224] The formulation (DP) used in the storage stability, accelerated stability, and stress stability tests was prepared by filling 0.4 mL of FDS into a 2 mL type 1 glass vial. The DP was incubated under several high temperature conditions compared to the storage temperature conditions. These accelerated conditions were selected to simulate the conditions to which the DP could be exposed during manufacturing and handling, and to elucidate the degradation pathways of co-formulated H1H17203P, H1H17139P, and H1H17161P. An overview of the storage conditions, accelerated conditions, and stress stability conditions for the co-formulated DP is presented in Table 12, and the analysis plan is presented in Table 13.

[0225] (Table 12) Stability investigation tests for the combined DP of H1H17203P, H1H17139P, and H1H17161P TIFF2025090691000012.tif64161 a Frozen at -30°C and thawed at room temperature.

[0226] (Table 13) Analysis plan for the stability investigation tests for the combined DP of H1H17203P, H1H17139P, and H1H17161P TIFF2025090691000013.tif105161

[0227] Storage stability investigation of co-formulated H1H17203P, H1H17139P, and H1H17161P DP (1:1:1, total protein 50 mg / mL) Storage stability investigation data for 3 months of co-formulated H1H17203P, H1H17139P, and H1H17161P DP are shown. The co-formulated H1H17203P, H1H17139P, and H1H17161P DP was physically stable when stored at 5°C for 3 months. When the co-formulated H1H17203P, H1H17139P, and H1H17161P DP (1:1:1, total protein 50 mg / mL) was stored at 5°C for 3 months, a 0.2% increase was observed in total HMW species by SE-UPLC, and after storage at 5°C for 18 months, a 0.6 increase in total HMW was observed by SE-UPLC. No perceptible changes were detected in the physical and chemical stabilities of the co-formulated H1H17203P, H1H17139P, and H1H17161P DP (1:1:1, total protein 50 mg / mL) in any of the other monitored characteristics. H1H17203P, H1H17139P, and H1H17161P maintained their potency over the 3-month evaluation period as determined by bioassay analysis. See Table 14.

[0228] Accelerated stability investigation of co-formulated H1H17203P, H1H17139P, and H1H17161P DP (1:1:1, total protein 50 mg / mL) The analysis results of the co-formulated H1H17203P, H1H17139P, and H1H17161P DPs after incubation under accelerated conditions are presented in 4. After incubation at 45°C for 28 days, a 1.3% increase (SE-UPLC) was observed in the relative amount of total HMW species. After incubation at 45°C for 28 days, increases of 14.7%, 14.4%, and 22.9% were observed in the acidic charge variant species (CEX-UPLC) of H1H17203P, H1H17139P, and H1H17161P, respectively. After incubation at 25°C for 3 months, increases of 2.6%, 2.5%, and 5.6% were observed in the acidic charge variant species (CEX-UPLC) of H1H17203P, H1H17139P, and H1H17161P, respectively. H1H17203P, H1H17139P, and H1H17161P maintained their potency when determined by bioassay analysis after incubation under accelerated conditions. From the results of the accelerated stability test, increases in the relative amounts of total HMW species, total LMW species, and acidic charge variants for H1H17203P, H1H17139P, and H1H17161P were shown to be the main degradation pathways of the co-formulated H1H17203P, H1H17139P, and H1H17161P DPs (1:1:1, 50 mg / mL total protein). See Table 15.

[0229] Stress stability investigation of co-formulated H1H17203P, H1H17139P, and H1H17161P DPs (1:1:1, 50 mg / mL total protein) The results of the stress stability tests of the co-formulated H1H17203P, H1H17139P, and H1H17161P DPs are presented in Table 16. The co-formulated H1H17203P, H1H17139P, and H1H17161P DPs were physically and chemically stable when stirred for 120 minutes (vortexed at ambient temperature) or when subjected to 8 freeze / thaw cycles (frozen at -30°C and thawed at room temperature). No perceivable changes in physical or chemical stability were detected in any of the monitored properties. H1H17203P, H1H17139P, and H1H17161P maintained their potency when stirred for 120 minutes or when exposed to 8 freeze / thaw cycles. See Table 16.

[0230] Conclusion of the Stability Investigation of the Co-Formulated H1H17203-3471-3479 Formulation From the results of the storage stability, accelerated stability, and stress stability tests, it is suggested that the co-formulated H1H17203P, H1H17139P, and H1H17161P DPs are stable during manufacturing (filling / finishing and labeling operations). The combined DP formulation of H1H17203P, H1H17139P, and H1H17161P can withstand short-term exposure to room temperature without compromising physical and chemical stability. The combined DP of H1H17203P, H1H17139P, and H1H17161P will be stored at 2°C to 8°C, and exposure to temperatures above 8°C will be limited.

[0231] The long-term storage of co-formulated H1H17203P, H1H17139P, and H1H17161P DPs at 2 - 8°C was determined over 36 months (Table 14). After storage at 5°C for 36 months, an increase of 0.7% was observed for each of the total HMW species and total LMW species. For H1H17203P, H1H17139P, and H1H17161P respectively, changes in acidic species of 1.1%, 1.2%, and 7.3% were observed. Over the 36-month period of the study, no significant change in potency was observed, suggesting that the anti-EBOV formulation provides acceptable long-term storage stability under refrigerated conditions.

[0232] (Table 14) Stability investigation of a combined DP of H1H17203P, H1H17139P, and H1H17161P (1:1:1, 50 mg / mL total protein) stored at 5°C TIFF2025090691000014.tif224170NR, not required.

[0233] The results from the accelerated tests are presented in Table 15 below. After incubation at 25°C for 3 months, an increase of 0.6% was observed in the relative amounts of total HMW species and total LMW species. For H1H17203P, H1H17139P, and H1H17161P respectively, increases in acidic charge variants of 2.6%, 2.5%, and 5.6% were observed. The co-formulated H1H17203P, H1H17139P, and H1H17161P DPs maintained their potency after incubation under accelerated conditions. After incubation at 45°C for 28 days, an increase of 1.3% was observed in the relative amount of total HMW species, while the increase in total LMW species was 1.9%.

[0234] From the results of the stress stability tests and accelerated stability tests on the co-formulated H1H17203P, H1H17139P, and H1H17161P DPs, it was shown that the increases in the relative amounts of total HMW species, total LMW species, and acidic charge variants were limited for the co-formulated H1H17203P, H1H17139P, and H1H17161P DPs, similar to the results in the individual formulations.

[0235] (Table 15) Stability investigation of the combination DP (1:1:1, 50 mg / mL total protein) of H1H17203P, H1H17139P, and H1H17161P - Influence of accelerated conditions TIFF2025090691000015.tif218167NR: Not required.

[0236] From the results of the stress stability tests, it was shown that the co-formulated DP of H1H17203P, H1H17139P, and H1H17161P was physically and chemically stable when stirred for 120 minutes (vortexed at room temperature) or when subjected to 8 freeze / thaw cycles (Table 16). No perceivable changes in physical or chemical stability were detected in any of the monitored characteristics. The potency of the co-formulated DP of H1H17203P, H1H17139P, and H1H17161P was maintained when stirred or when exposed to freeze / thaw cycles.

[0237] (Table 16) Stability investigation of the combination DP (1:1:1, 50 mg / mL total protein) of H1H17203P, H1H17139P, and H1H17161P - Influence of stress conditions TIFF2025090691000016.tif230170NR, Not required

[0238] Conclusion The co-formulated DP of H1H17203P, H1H17139P, and H1H17161P is manufactured as a liquid DP for IV administration. The DP of H1H17203P, H1H17139P, and H1H17161P may contain 16.7 mg / mL H1H17203P, 16.7 mg / mL H1H17139P, and 16.7 mg / mL H1H17161P formulated in a solution containing 10 mM histidine, pH 6.0, 0.1% (w / v) polysorbate 80, and 10% (w / v) sucrose. Based on the test results herein:

[0239] The DPs of co-formulated H1H17203P, H1H17139P, and H1H17161P are stable when stored at 2 - 8°C for at least 3 months.

[0240] The major degradation pathways identified under accelerated conditions were the formation of HMW and LMW species, as well as acidic charge variants.

[0241] Example 6: Influence of Different Buffers and pH The influence of buffers and pH on the thermal stability of three anti-EBOV antibodies was verified in liquid formulations by incubating 100 mg / mL total antibody at 45°C for 28 days in a series of buffer systems over various pH ranges. The following pH and buffer systems were tested: citrate (pH 5.2, 6.0, 6.8) and histidine (pH 5.2, 6.0, 6.8). Based on the results of SE-UPLC analysis, maximum protein stability was observed when formulating the antibody at pH 6.0 in histidine buffer (Tables 17 - 20 and 22 - 25). The presence of subvisible particles was determined using micro fluid imaging (MFI) in three dosage form antibody formulations (Table 21).

[0242] (Table 17) Influence of Buffer and pH on the Stability of 100 mg / mL H1H17203P Incubated at 45°C for 28 Days - Results of Visual, OD, pH, SE-UPLC, and RP-UPLC TIFF2025090691000017.tif17516210 mM citrate, pH 5.2, 28-day sample was not run on the UPLC instrument due to visual failure

[0243] (Table 18) Influence of Buffer and pH on the Stability of 100 mg / mL H1H17139P Incubated at 45°C for 28 Days - Results of Visual, OD, pH, SE-UPLC, and RP-UPLC TIFF2025090691000018.tif175162

[0244] (Table 19) Effects of buffer and pH on the stability of 100 mg / mL H1H17161P incubated at 45°C for 28 days - Results of visual, OD, pH, SE-UPLC, and RP-UPLC TIFF2025090691000019.tif175162

[0245] (Table 20) Effects of buffer and pH on the stability of the combination of 100 mg / mL H1H17203P, H1H17139P, and H1H17161P incubated at 45°C for 28 days - Results of visual, OD, pH, SE-UPLC, and RP-UPLC TIFF2025090691000020.tif180163

[0246] (Table 21) Results of MFI for the combined formulation of H1H17203P, H1H17139P, and H1H17161P TIFF2025090691000021.tif131140

[0247] Based on the results of CEX-UPLC analysis, maximum protein stability was observed when the antibody was formulated in histidine buffer at pH 5.2 - 6.8 or in acetate buffer at pH 5.2 - 6.8. These analyses also revealed that aggregation (i.e., formation of HMW species), fragmentation (i.e., formation of LMW species), and formation of charge variants were the major degradation pathways. Histidine buffer was selected as the formulation buffer because it provided the overall best level of protein stabilization with respect to the formation of HMW and LMW species and charge variants. pH 6.0 was selected for the formulation because the formation of HMW species and charge variants, which are the major degradation pathways, was minimized at this pH. Based on these results, 10 mM histidine buffer at pH 6.0 was selected for the individual and combined formulations against EBOV. See Tables 22 - 25.

[0248] (Table 22) Effects of buffer and pH on the stability of 100 mg / mL H1H17203P incubated at 45 °C for 28 days - Results of CEX-UPLC TIFF2025090691000022.tif172128 The citrate, pH 5.2, 28-day sample was not run on the UPLC instrument due to visual failure.

[0249] (Table 23) Effects of buffer and pH on the stability of 100 mg / mL H1H17139P incubated at 45 °C for 28 days - Results of CEX-UPLC TIFF2025090691000023.tif172128

[0250] (Table 24) Effects of buffer and pH on the stability of 100 mg / mL H1H17161P incubated at 45 °C for 28 days - Results of CEX-UPLC TIFF2025090691000024.tif172128

[0251] (Table 25) Effects of buffer and pH on the stability of the combination of 100 mg / mL H1H17203P, H1H17139P, and H1H17161P incubated at 45 °C for 28 days - Results of CEX-UPLC TIFF2025090691000025.tif176169

[0252] Example 7: Selection of stabilizers under stress conditions From Example 6, pH 6.0 and 10 mM L-histidine were identified as the optimal buffer systems for the individually formulated antibody and the three-formulation co-formulation at 100 mg / mL. Additional excipients such as heat stabilizers, surfactants, and antioxidants were evaluated in this example for their effects on forced degradation and stability. Stabilizers such as sucrose, surfactants, proline, or methionine are often added to antibody formulations to enhance the thermal stability of proteins in liquid formulations. Separately or in combination, anti-EBOV antibodies formulated in liquid formulations showed improved stability under accelerated conditions and equivalent stability results under vortex stress when formulated with 5% w / v sucrose and 0.1% w / v polysorbate 80 or 0.1% polysorbate 20. (Tables 27 - 32). Table 26 provides the excipient variables of Tables 16 - 21 and each formulation. The basic formulation contains 33.3 mg / mL anti-EBOV antibody, 5% w / v sucrose, and 10 mM histidine.

[0253] (Table 26) Excipients and formulation components tested TIFF2025090691000026.tif78167

[0254] (Table 27) Effect of excipients on the stability of 33 mg / mL H1H17203P antibody incubated at 40°C for up to 2 months - Visual, OD, pH, SE-UPLC, RP-UPLC concentration, CEX-UPLC TIFF2025090691000027.tif223166TIFF2025090691000028.tif213166

[0255] (Table 28) Effect of excipients on the stability of 33 mg / mL H1H17139P antibody incubated at 40°C for up to 2 months - Visual, OD, pH, SE-UPLC, RP-UPLC concentration, CEX-UPLC TIFF2025090691000029.tif223164TIFF2025090691000030.tif213164

[0256] (Table 29) Effect of excipients on the stability of 33 mg / mL H1H17161P antibody incubated at 40°C for up to 2 months - Visual, OD, pH, SE-UPLC, RP-UPLC concentration, CEX-UPLC TIFF2025090691000031.tif224164TIFF2025090691000032.tif213164

[0257] (Table 30) Effect of excipients on the stability of 100 mg / mL H1H17203P, H1H17139P, and H1H17161P antibodies (1:1:1 ratio) incubated at 40°C for up to 2 months - Visual, OD, pH, SE-UPLC, RP-UPLC concentration TIFF2025090691000033.tif213163TIFF2025090691000034.tif224163TIFF2025090691000035.tif168163

[0258] (Table 31) Effect of excipients on the stability of 100 mg / mL H1H17203P, H1H17139P, and H1H17161P antibodies (1:1:1 ratio) incubated at 40°C for up to 2 months - CEX-UPLC TIFF2025090691000036.tif218166TIFF2025090691000037.tif224166TIFF2025090691000038.tif86166

[0259] (Table 32) Effect of excipients on the stability of 100 mg / mL H1H17203P, H1H17139P, and H1H17161P antibodies (1:1:1 ratio) incubated at 40°C for up to 2 months - MFI results TIFF2025090691000039.tif217138

[0260] Example 8: Selection of organic co-solvents for agitation stress For example, stabilizers such as surfactants and organic co-solvents are often added to antibody formulations to protect proteins from agitation-induced aggregation. The effects of organic co-solvents and surfactants on the agitation stress stability and thermal stability of individual anti-EBOV antibodies at 33.3 mg / mL and co-formulated cocktails of three antibodies at 100 mg / mL (in a 1:1:1 ratio) were verified in liquid formulations. From the previous examples, polysorbate 80 was identified as an important excipient component. This example evaluates polysorbate 80 at different % w / g under agitation stress induced by orbital shaking at 250 rpm for 48 hours. Refer to Tables 33 to 40. The basic formulation contains 33.3 mg / mL of a single antibody (or 100 mg / mL of total antibody in co-formulation), 10 mM histidine, pH 6.0, 5% w / v sucrose, along with 0.04% to 0.2% polysorbate 80 as shown. Formulations without surfactants have lower stability under agitation stress compared to formulations containing PS80 at 0.04% w / v or higher. Furthermore, among formulations containing polysorbate 80 at 0.04% w / v or higher, there was no significant difference in the results of MFI and SE-UPLC.

[0261] (Table 33) Effect of polysorbate 80 on agitation stress in formulations containing H1H17203P anti-EBOV - Visual, OD, pH, HMW%, native%, LMW%, and PS80 %w / v concentration TIFF2025090691000040.tif69165

[0262] (Table 34) Effect of polysorbate 80 on agitation stress in formulations containing H1H17139P anti-EBOV - Visual, OD, pH, HMW%, native%, LMW%, and PS80 %w / v concentration TIFF2025090691000041.tif69161

[0263] (Table 35) Effect of polysorbate 80 on agitation stress in formulations containing H1H17161P anti-EBOV - Visual, OD, pH, HMW%, native%, LMW%, and PS80 %w / v concentration TIFF2025090691000042.tif69161

[0264] (Table 36) Effect of polysorbate 80 on agitation stress in formulations containing H1H17203P, H1H17139P, and H1H17161P anti-EBOV - Visual, OD, pH, HMW%, Unmodified%, LMW%, and PS80 %w / v concentration TIFF2025090691000043.tif64161

[0265] (Table 37) Effect of polysorbate 80 on agitation stress in formulations containing H1H17203P anti-EBOV - MFI results TIFF2025090691000044.tif90128

[0266] (Table 38) Effect of polysorbate 80 on agitation stress in formulations containing H1H17139P anti-EBOV - MFI results TIFF2025090691000045.tif90128

[0267] (Table 39) Effect of polysorbate 80 on agitation stress in formulations containing H1H17161P anti-EBOV - MFI results TIFF2025090691000046.tif90128

[0268] (Table 40) Effect of polysorbate 80 on agitation stress in formulations containing H1H17203P, H1H17139P, and H1H17161P anti-EBOV - MFI results TIFF2025090691000047.tif102128

[0269] The anti-EBOV antibody was unstable when stirred by orbital shaking at 250 rpm in the absence of an organic co-solvent or surfactant. After stirring by orbital shaking at 250 rpm in the absence of a co-solvent or surfactant, the solution became turbid, the turbidity increased significantly, aggregates increased as determined by SE-UPLC, and there was a decrease in protein recovery by RP-UPLC. In contrast, 0.1% polysorbate 80 protected the antibody from agitation-induced instability.

[0270] Example 9: Selection of Stabilizer Concentration The objective of this example was to identify the levels of stabilizer components that could be used in the development of a formulation recipe to support an individual anti-EBOV antibody at 33.3 mg / mL and a three-dosage form antibody co-formulation cocktail (1:1:1 ratio) at 100 mg / mL. 10% sucrose was selected for the initial formulation and tested at the following various amounts to evaluate any changes in antibody stability: 5%, 10%, 15%, and 20%. The basic formulation contained 100 mg / mL antibody, 10 mM histidine, pH 6.0, and 0.1% polysorbate 80.

[0271] During the development of the low-concentration formulation, sucrose was selected as a heat stabilizer for the anti-EBOV antibody. For the development of the high-concentration formulation, various concentrations of sucrose were evaluated for the stability of the antibody at a concentration of 100 mg / mL at 25 °C for 0 - 6 months and at 40 °C for 0 - 3 months. When the formulation was incubated at 40 °C for 28 days, the formation of HMW species decreased with increasing sucrose concentration.

[0272] This test showed that the protein stability was equivalent in formulations containing 10% w / v sucrose compared to formulations containing 15% or 20% sucrose, and thus additional sucrose above 10% w / v was not necessary. See Tables 41 - 48.

[0273] (Table 41) Accelerated Stability of High-Concentration (100 mg / mL) H1H17203P Anti-EBOV Antibody with Heat Stabilizer - Visual, OD, and pH TIFF2025090691000048.tif197115

[0274] (Table 42) Accelerated stability of high-concentration (100 mg / mL) H1H17203P anti-EBOV antibody using heat stabilizers - Results of SE-UPLC, RP-UPLC, and CEX-UPLC TIFF2025090691000049.tif216166

[0275] (Table 43) Accelerated stability of high-concentration (100 mg / mL) H1H17139P anti-EBOV antibody using heat stabilizers - Visual, OD, and pH TIFF2025090691000050.tif196117

[0276] (Table 44) Accelerated stability of high-concentration (100 mg / mL) H1H17139P anti-EBOV antibody using heat stabilizers - Results of SE-UPLC, RP-UPLC, and CEX-UPLC TIFF2025090691000051.tif216166

[0277] (Table 45) Accelerated stability of high-concentration (100 mg / mL) H1H17161P anti-EBOV antibody using heat stabilizers - Visual, OD and pH TIFF2025090691000052.tif188128

[0278] (Table 46) Accelerated stability of high-concentration (100 mg / mL) H1H17161P anti-EBOV antibody using heat stabilizers - Results of SE-UPLC, RP-UPLC, and CEX-UPLC TIFF2025090691000053.tif198165

[0279] (Table 47) Accelerated stability of high-concentration H1H17203P, H1H17139P, and H1H17161P anti-EBOV antibodies using heat stabilizers - Visual, OD, pH, SE-UPLC, and RP-UPLC TIFF2025090691000054.tif245167

[0280] (Table 48) Accelerated stability of anti-EBOV antibodies of H1H17203P, H1H17139P, and H1H17161P at high concentration (100 mg / mL) using heat stabilizer - Results of CEX-UPLC TIFF2025090691000055.tif250166

[0281] Example 10: Stability investigation test (12 months) of formulated drug substances (FDS: Formulated Drug Substance) for individually formulated H1H17203P C2P1 FDS, H1H17139P C2P1 FDS, and H1H17161P C2P1 FDS The research FDS and clinical FDS are manufactured using the same process, and the methods used to evaluate the stability of research materials are the same as those used to evaluate the stability of materials intended for clinical trials. Therefore, the results obtained from the stability investigation test can be used to evaluate the stability of materials intended for clinical trials.

[0282] The long-term storage stability test of the present invention evaluated the stability of H1H17203P C2P1 FDS, H1H17139P C2P1 FDS, and H1H17161P C2P1 FDS through 12 months of storage at -80°C and -30°C. When stored at -80°C and -30°C, H1H17203P C2P1 FDS, H1H17139P C2P1 FDS, and H1H17161P C2P1 FDS are stable for at least 12 months.

[0283] As presented in Table 49, the ongoing long-term stability test continues for the duration of the stability protocol. Samples were analyzed according to the analytical plan outlined in Table 50. Present the analytical plan, preliminary acceptance criteria, and sampling plan in Table 50.

[0284] (Table 49) Summary of formulated drug substance stability tests for H1H17203P, H1H17139P, and H1H17161P TIFF2025090691000056.tif156167RH, Relative Humidity

[0285] (Table 50) Analytical Plan and Preliminary Pass / Fail Criteria for the Stability Investigation Test of Formulated Drug Substances H1H17203P, H1H17139P, and H1H17161P TIFF2025090691000057.tif216166280 nm A 280 Absorbance; CEX, Cation Exchange Chromatography; SE-UPLC, Size Exclusion Ultra-High Performance Liquid Chromatography; HMW, High Molecular Weight; HC, Heavy Chain; LC, Light Chain; LMW, Low Molecular Weight; MCE, Microchip Capillary Electrophoresis; NGHC, Non-Glycosylated Heavy Chain

[0286] Results of the Stability Test of Formulated Drug Substances Results of the Long-Term Storage Stability Investigation Test of H1H17203P FDS In the stability investigation test, C2P1 FDS was found to be physically and chemically stable when stored at -80 °C and -30 °C (Tables 51 and 52, respectively). No perceivable changes in physical or chemical stability were detected in any of the monitored characteristics. The stability investigation test to verify C2P1 FDS under long-term storage conditions will continue for 84 months.

[0287] Results of the Accelerated Stability Test of H1H17203P FDS From the stability investigation tests, C2P1 FDS was shown to be physically and chemically stable when stored at -20 °C for 3 months and at 5 °C for 56 days (Tables 53 and 54). No perceivable changes in physical or chemical stability were detected in any of the monitored properties. For H1H17203P, a slight increase of 0.3% in HMW species was observed by SE-UPLC after storage at 25 °C / 60% RH (Table 54) for 28 days. No significant changes were observed in the other monitored properties after incubation at 25 °C / 60% RH for 28 days. Test investigations to verify C2P1 FDS were completed under accelerated storage conditions of -20 °C, 5 °C, and 25 °C / 60% RH.

[0288] Results of the stress stability test of H1H17203P FDS The results of the stability investigation tests from the analysis of C2P1 FDS under stress conditions of 40 °C / 75% RH are presented in Table 55. An increase of 1.2% in HMW species and an increase of 0.4% in LMW species were observed by SE-UPLC. An increase of 11.1% in Region 1 was observed by CEX-UPLC. No significant changes were observed in the other monitored properties. C2P1 FDS maintained its potency after incubation at 40 °C / 75% RH for 28 days. Test investigations to verify C2P1 FDS were completed under stress storage conditions of 40 °C / 75% RH.

[0289] The results of the stability investigation from the analysis of C2P1 FDS after agitation and freeze / thaw conditions are presented in Table 56. C2P1 FDS was physically and chemically stable when agitated for 120 minutes (vortexed at ambient temperature) or when subjected to 8 freeze / thaw cycles (-30 °C for freezing and room temperature for thawing). No perceivable changes in the physical or chemical stability of H1H17203P were observed in any of the monitored properties. Test investigations to verify C2P1 FDS were completed under agitation and freeze / thaw conditions.

[0290] Results of the long-term storage stability investigation test of H1H17139P FDS In the stability investigation test, C2P1 FDS was found to be physically and chemically stable when stored at -80 °C and -30 °C (Tables 57 and 58). No perceivable changes in physical or chemical stability were detected in any of the monitored characteristics. The stability investigation test to verify C2P1 FDS under long-term storage conditions continues for 84 months.

[0291] Results of the accelerated stability test of H1H17139P FDS From the stability investigation test, C2P1 FDS was shown to be physically and chemically stable when stored at -20 °C for 3 months and at 5 °C for 56 days (Tables 59 and 60). No perceivable changes in physical or chemical stability were detected in any of the monitored characteristics. After storage at 25 °C / 60% RH (Table 60) for 28 days, a slight increase of 0.3% was observed in the HMW species by SE-UPLC. After incubation at 25 °C / 60% RH for 28 days, no significant changes were observed in the other monitored characteristics. The test investigation to verify C2P1 FDS under accelerated storage conditions of -20 °C, 5 °C, and 25 °C / 60% RH was completed.

[0292] Results of the stress stability test of H1H17139P FDS The results of the stability investigation test from the analysis of C2P1 FDS under stress conditions of 40 °C / 75% RH are presented in Table 61. An increase of 0.9% was observed in the HMW species and an increase of 0.4% was observed in the LMW species by SE-UPLC. An increase of 11.1% in Region 1 and a decrease of 12.9% in Region 2 were observed by CEX-UPLC. No significant changes were observed in the other monitored characteristics. C2P1 FDS maintained its potency after incubation at 40 °C / 75% RH for 28 days. The test investigation to verify C2P1 FDS under stress storage conditions of 40 °C / 75% RH was completed.

[0293] After the stirring and freezing / thawing conditions, the results of the stability investigation from the analysis of C2P1 FDS are presented in Table 62. C2P1 FDS was physically and chemically stable when stirred for 120 minutes (vortexed at ambient temperature) or when subjected to 8 freeze / thaw cycles (-30 °C for freezing and room temperature for thawing). No perceivable changes in the physical or chemical stability of H1H17139P were observed in any of the monitored characteristics. A test investigation to verify C2P1 FDS under stirring and freezing / thawing conditions was completed.

[0294] Results of the long-term storage stability investigation test of H1H17161P FDS In the stability investigation test, C2P1 FDS was found to be physically and chemically stable when stored at -80 °C and -30 °C (Tables 63 and 64). No perceivable changes in the physical or chemical stability were detected in any of the monitored characteristics. The stability investigation test to verify C2P1 FDS under long-term storage conditions continues for 84 months.

[0295] Results of the accelerated stability test of H1H17161P FDS From the stability investigation test, C2P1 FDS was shown to be physically and chemically stable when stored at -20 °C for 3 months and at 5 °C for 56 days (Tables 65 and 66). No perceivable changes in the physical or chemical stability were detected in any of the monitored characteristics. After storage at 25 °C / 60% RH (Table 66) for 28 days, a slight increase of 0.4% in the HMW species was observed by SE-UPLC. No significant changes were observed in the other monitored characteristics after incubation at 25 °C / 60% RH for 28 days. A test investigation to verify C2P1 FDS under accelerated storage conditions of -20 °C, 5 °C, and 25 °C / 60% RH was completed.

[0296] Results of the stress stability test of H1H17161P FDS The results of the stability investigation test from the analysis of C2P1 FDS under stress conditions of 40°C / 75%RH are presented in Table 67. An increase of 1.0% was observed for the HMW species and an increase of 0.4% was observed for the LMW species by SE-UPLC. An increase of 14.4% was observed in Region 1 by CEX-UPLC. No significant changes were observed in other monitored characteristics. C2P1 FDS maintained its potency after incubation at 40°C / 75%RH for 28 days. The test investigation to verify C2P1 FDS under stress storage conditions of 40°C / 75%RH was completed.

[0297] The results of the stability investigation from the analysis of C2P1 FDS after stirring and freezing / thawing conditions are presented in Table 68. C2P1 FDS was physically and chemically stable when stirred for 120 minutes (vortexed at ambient temperature) or when subjected to 8 freeze / thaw cycles (frozen at -30°C and thawed at room temperature). No perceivable changes in the physical or chemical stability of H1H17161P were observed in any of the monitored characteristics. The test investigation to verify C2P1 FDS under stirring and freezing / thawing conditions was completed.

[0298] Conclusion on Stability H1H17203P C2P1 FDS, H1H17139P C2P1 FDS, and H1H17161P C2P1 FDS can each withstand short-term exposure to refrigeration, room temperature, and agitation stress without compromising the physical or chemical stability of the protein and can be frozen and thawed. These results indicate that H1H17203P C2P1 FDS, H1H17139P C2P1 FDS, and H1H17161P C2P1 FDS are stable during the manufacture of the combination formulation (DP) of the three dosage forms. Exposure of H1H17203P C2P1 FDS, H1H17139P C2P1 FDS, and H1H17161P C2P1 FDS to temperatures above room temperature should be avoided.

[0299] Recommended Storage Conditions The recommended long-term storage temperature for H1H17203P C2P1 FDS, H1H17139P C2P1 FDS, and H1H17161P C2P1 FDS is -30°C.

[0300] (Table 51) Stability investigation of 100 mg / mL H1H17203P C2P1 formulated drug substance stored at -80°C TIFF2025090691000058.tif142160CEX, cation exchange; FDS, formulated drug substance; HDPE, high-density polyethylene; HMW, high molecular weight; LEFVP, liquid essentially free of visible particles; LMW, low molecular weight; MCE, microchip capillary electrophoresis; NGHC, non-glycosylated heavy chain; NR, not required by protocol; RP, reverse phase; SE, size exclusion; UPLC, ultra-high performance liquid chromatography; ADCC: antibody-dependent cell-mediated cytotoxicity activity;

[0301] (Table 52) Stability investigation of 100 mg / mL H1H17203P C2P1 formulated drug substance stored at -30°C TIFF2025090691000059.tif142160CEX, cation exchange; FDS, formulated drug substance; FDG, formulation development group; HDPE, high-density polyethylene; HMW, high molecular weight; LEFVP, liquid essentially free of visible particles; LMW, low molecular weight; MCE, microchip capillary electrophoresis; NGHC, non-glycosylated heavy chain; NR, not required by protocol; OD, optical density; RP, reverse phase; SE, size exclusion; UPLC, ultra-high performance liquid chromatography; ADCC: antibody-dependent cell-mediated cytotoxicity activity;

[0302] (Table 53) Stability investigation of 100 mg / mL H1H17203P C2P1 formulated drug substance - Influence of accelerated conditions at -20°C TIFF2025090691000060.tif142160CEX, Cation Exchange; FDS, Formulated Drug Substance; FDG, Formulation Development Group; HDPE, High-Density Polyethylene; HMW, High Molecular Weight; LEFVP, Liquid Essentially Free of Visible Particles; LMW, Low Molecular Weight; MCE, Microchip Capillary Electrophoresis; NGHC, Non-Glycosylated Heavy Chain; NR, Not Required by Protocol; OD, Optical Density; RP, Reverse Phase; SE, Size Exclusion; UPLC, Ultra Performance Liquid Chromatography; ADCC: Antibody-Dependent Cellular Cytotoxicity Activity;

[0303] (Table 54) Stability Investigation of 100 mg / mL H1H17203P C2P1 Formulated Drug Substance - Influence of Accelerated Conditions TIFF2025090691000061.tif160160CEX, Cation Exchange; FDS, Formulated Drug Substance; FDG, Formulation Development Group; HDPE, High-Density Polyethylene; HMW, High Molecular Weight; LEFVP, Liquid Essentially Free of Visible Particles; LMW, Low Molecular Weight; MCE, Microchip Capillary Electrophoresis; NGHC, Non-Glycosylated Heavy Chain; NR, Not Required by Protocol; OD, Optical Density; RP, Reverse Phase; SE, Size Exclusion; UPLC, Ultra Performance Liquid Chromatography; ADCC: Antibody-Dependent Cellular Cytotoxicity Activity;

[0304] (Table 55) Stability Investigation of 100 mg / mL H1H17203P C2P1 Formulated Drug Substance - Influence of Stress Conditions TIFF2025090691000062.tif140158CEX, Cation Exchange; FDS, Formulated Drug Substance; FDG, Formulation Development Group; HDPE, High-Density Polyethylene; HMW, High Molecular Weight; LEFVP, Liquid Essentially Free of Visible Particles; LMW, Low Molecular Weight; MCE, Microchip Capillary Electrophoresis; NGHC, Non-Glycosylated Heavy Chain; NR, Not Required by Protocol; OD, Optical Density; RP, Reverse Phase; SE, Size Exclusion; UPLC, Ultra Performance Liquid Chromatography; ADCC: Antibody-Dependent Cellular Cytotoxicity Activity;

[0305] (Table 56) Stability Investigation of 100 mg / mL H1H17203P C2P1 Formulated Drug Substance - Influence of Stirring and Freeze-Thaw TIFF2025090691000063.tif145158CEX, Cation Exchange; FDS, Formulated Drug Substance; FDG, Formulation Development Group; HDPE, High-Density Polyethylene; HMW, High Molecular Weight; LEFVP, Liquid Essentially Free of Visible Particles; LMW, Low Molecular Weight; MCE, Microchip Capillary Electrophoresis; NGHC, Non-Glycosylated Heavy Chain; NR, Not Required by Protocol; OD, Optical Density; RP, Reverse Phase; SE, Size Exclusion; UPLC, Ultra-High Performance Liquid Chromatography; ADCC: Antibody-Dependent Cell-Mediated Cytotoxicity Activity;

[0306] (Table 57) Stability Investigation of 100 mg / mL H1H17139P C2P1 Formulated Drug Substance Stored at -80°C TIFF2025090691000064.tif135158CEX, Cation Exchange; FDS, Formulated Drug Substance; FDG, Formulation Development Group; HDPE, High-Density Polyethylene; HMW, High Molecular Weight; LEFVP, Liquid Essentially Free of Visible Particles; LMW, Low Molecular Weight; MCE, Microchip Capillary Electrophoresis; NGHC, Non-Glycosylated Heavy Chain; NR, Not Required by Protocol; OD, Optical Density; RP, Reverse Phase; SE, Size Exclusion; UPLC, Ultra-High Performance Liquid Chromatography; ADCC: Antibody-Dependent Cell-Mediated Cytotoxicity Activity;

[0307] (Table 58) Stability Investigation of 100 mg / mL H1H17139P C2P1 Formulated Drug Substance Stored at -30°C TIFF2025090691000065.tif127158CEX, Cation Exchange; FDS, Formulated Drug Substance; FDG, Formulation Development Group; HDPE, High-Density Polyethylene; HMW, High Molecular Weight; LEFVP, Liquid Essentially Free of Visible Particles; LMW, Low Molecular Weight; MCE, Microchip Capillary Electrophoresis; NGHC, Non-Glycosylated Heavy Chain; NR, Not Required by Protocol; OD, Optical Density; RP, Reverse Phase; SE, Size Exclusion; UPLC, Ultra-High Performance Liquid Chromatography; ADCC: Antibody-Dependent Cell-Mediated Cytotoxicity Activity;

[0308] (Table 59) Stability Investigation of 100 mg / mL H1H17139P C2P1 Formulated Drug Substance - Influence of Accelerated Conditions at -20°C TIFF2025090691000066.tif137158CEX, Cation Exchange; FDS, Formulated Drug Substance; FDG, Formulation Development Group; HDPE, High-Density Polyethylene; HMW, High Molecular Weight; LEFVP, Liquid Essentially Free of Visible Particles; LMW, Low Molecular Weight; MCE, Microchip Capillary Electrophoresis; NGHC, Non-Glycosylated Heavy Chain; NR, Not Required by Protocol; OD, Optical Density; RP, Reverse Phase; SE, Size Exclusion; UPLC, Ultra Performance Liquid Chromatography; ADCC: Antibody-Dependent Cellular Cytotoxicity Activity;

[0309] (Table 60) Stability Investigation of 100 mg / mL H1H17139P C2P1 Formulated Drug Substance - Influence of Accelerated Conditions TIFF2025090691000067.tif135158CEX, Cation Exchange; FDS, Formulated Drug Substance; FDG, Formulation Development Group; HDPE, High-Density Polyethylene; HMW, High Molecular Weight; LEFVP, Liquid Essentially Free of Visible Particles; LMW, Low Molecular Weight; MCE, Microchip Capillary Electrophoresis; NGHC, Non-Glycosylated Heavy Chain; NR, Not Required by Protocol; OD, Optical Density; RP, Reverse Phase; SE, Size Exclusion; UPLC, Ultra Performance Liquid Chromatography; ADCC: Antibody-Dependent Cellular Cytotoxicity Activity;

[0310] (Table 61) Stability Investigation of 100 mg / mL H1H17139P C2P1 Formulated Drug Substance - Influence of Stress Conditions TIFF2025090691000068.tif131158CEX, Cation Exchange; FDS, Formulated Drug Substance; FDG, Formulation Development Group; HDPE, High-Density Polyethylene; HMW, High Molecular Weight; LEFVP, Liquid Essentially Free of Visible Particles; LMW, Low Molecular Weight; MCE, Microchip Capillary Electrophoresis; NGHC, Non-Glycosylated Heavy Chain; NR, Not Required by Protocol; OD, Optical Density; RP, Reverse Phase; SE, Size Exclusion; UPLC, Ultra Performance Liquid Chromatography; ADCC: Antibody-Dependent Cellular Cytotoxicity Activity;

[0311] (Table 62) Stability Investigation of 100 mg / mL H1H17139P C2P1 Formulated Drug Substance - Influence of Stirring and Freeze-Thaw TIFF2025090691000069.tif135158CEX, Cation Exchange; FDS, Formulated Drug Substance; FDG, Formulation Development Group; HDPE, High-Density Polyethylene; HMW, High Molecular Weight; LEFVP, Liquid Essentially Free of Visible Particles; LMW, Low Molecular Weight; MCE, Microchip Capillary Electrophoresis; NGHC, Non-Glycosylated Heavy Chain; NR, Not Required by Protocol; OD, Optical Density; RP, Reverse Phase; SE, Size Exclusion; UPLC, Ultra-High Performance Liquid Chromatography; ADCC: Antibody-Dependent Cellular Cytotoxicity Activity;

[0312] (Table 63) Stability Investigation of 100 mg / mL H1H17161P C2P1 Formulated Drug Substance Stored at -80°C TIFF2025090691000070.tif145158CEX, Cation Exchange; FDS, Formulated Drug Substance; FDG, Formulation Development Group; HDPE, High-Density Polyethylene; HMW, High Molecular Weight; LEFVP, Liquid Essentially Free of Visible Particles; LMW, Low Molecular Weight; MCE, Microchip Capillary Electrophoresis; NGHC, Non-Glycosylated Heavy Chain; NR, Not Required by Protocol; OD, Optical Density; RP, Reverse Phase; SE, Size Exclusion; UPLC, Ultra-High Performance Liquid Chromatography

[0313] (Table 64) Stability Investigation of 100 mg / mL H1H17161P C2P1 Formulated Drug Substance Stored at -30°C TIFF2025090691000071.tif140158CEX, Cation Exchange; FDS, Formulated Drug Substance; FDG, Formulation Development Group; HDPE, High-Density Polyethylene; HMW, High Molecular Weight; LEFVP, Liquid Essentially Free of Visible Particles; LMW, Low Molecular Weight; MCE, Microchip Capillary Electrophoresis; NGHC, Non-Glycosylated Heavy Chain; NR, Not Required by Protocol; OD, Optical Density; RP, Reverse Phase; SE, Size Exclusion; UPLC, Ultra-High Performance Liquid Chromatography

[0314] (Table 65) Stability Investigation of 100 mg / mL H1H17161P C2P1 Formulated Drug Substance - Influence of Accelerated Conditions at -20°C TIFF2025090691000072.tif132158CEX, Cation Exchange; FDS, Formulated Drug Substance; FDG, Formulation Development Group; HDPE, High-Density Polyethylene; HMW, High Molecular Weight; LEFVP, Liquid Essentially Free of Visible Particles; LMW, Low Molecular Weight; MCE, Microchip Capillary Electrophoresis; NGHC, Non-Glycosylated Heavy Chain; NR, Not Required by Protocol; OD, Optical Density; RP, Reverse Phase; SE, Size Exclusion; UPLC, Ultra-High Performance Liquid Chromatography

[0315] (Table 66) Stability Investigation of 100 mg / mL H1H17161P C2P1 Formulated Drug Substance - Influence of Accelerated Conditions TIFF2025090691000073.tif149158CEX, Cation Exchange; FDS, Formulated Drug Substance; FDG, Formulation Development Group; HDPE, High-Density Polyethylene; HMW, High Molecular Weight; LEFVP, Liquid Essentially Free of Visible Particles; LMW, Low Molecular Weight; MCE, Microchip Capillary Electrophoresis; NGHC, Non-Glycosylated Heavy Chain; NR, Not Required by Protocol; OD, Optical Density; RP, Reverse Phase; SE, Size Exclusion; UPLC, Ultra-High Performance Liquid Chromatography

[0316] (Table 67) Stability Investigation of 100 mg / mL H1H17161P C2P1 Formulated Drug Substance - Influence of Stress Conditions TIFF2025090691000074.tif136158CEX, Cation Exchange; FDS, Formulated Drug Substance; FDG, Formulation Development Group; HDPE, High-Density Polyethylene; HMW, High Molecular Weight; LEFVP, Liquid Essentially Free of Visible Particles; LMW, Low Molecular Weight; MCE, Microchip Capillary Electrophoresis; NGHC, Non-Glycosylated Heavy Chain; NR, Not Required by Protocol; OD, Optical Density; RP, Reverse Phase; SE, Size Exclusion; UPLC, Ultra-High Performance Liquid Chromatography

[0317] (Table 68) Stability Investigation of 100 mg / mL H1H17161P C2P1 Formulated Drug Substance - Influence of Stirring and Freeze-Thaw TIFF2025090691000075.tif140158CEX, Cation Exchange; FDS, Formulated Drug Substance; FDG, Formulation Development Group; HDPE, High-Density Polyethylene; HMW, High Molecular Weight; LEFVP, Liquid Essentially Free of Visible Particles; LMW, Low Molecular Weight; MCE, Microchip Capillary Electrophoresis; NGHC, Non-Glycosylated Heavy Chain; NR, Not Required by Protocol; RP, Reverse Phase; SE, Size Exclusion; UPLC, Ultra-High Performance Liquid Chromatography

[0318] Example 11: Formulated Drug Substance Stability Investigation Test for Co-Formulated H1H17203P, H1H17139P, and H1H17161P C2P1 DP Stability investigation tests were conducted to evaluate the long-term storage conditions, accelerated conditions, and stress conditions for the manufactured combination C2P1 DP for research and development purposes. Six-month stability investigation data are available at accelerated conditions of 25°C / 60% RH and long-term storage conditions of 5°C for 6 months. Tests to verify 3 months of stress conditions at 40°C / 75% RH, agitation, and freeze-thaw have been completed.

[0319] Table 69 describes all the stability tests to verify the combination C2P1 DP. The analytical plan, preliminary pass / fail criteria, and sampling plan for the stability samples are presented in Table 70. As presented in Table 69, the stability tests continue over the entire duration of the stability protocol. Samples were analyzed according to the analytical plan outlined in Table 70.

[0320] (Table 69) Summary of Stability Tests for Combination Formulations TIFF2025090691000076.tif53156DP, Formulation; USP, United States Pharmacopeia; EP, European Pharmacopeia; RH, Relative Humidity

[0321] (Table 70) Analytical Plan for Formulation Stability Investigation Tests TIFF2025090691000077.tif229168A 280, Absorbance at 280 nm; CEX, Cation Exchange Chromatography; EU, Endotoxin Unit; HC, Heavy Chain; HMW, High Molecular Weight; LC, Light Chain; LMW, Low Molecular Weight; MCE, Microchip Capillary Electrophoresis; MFI, Micro-Flow Imaging (trademark); N / A, Not Applicable; NGHC, Non-Glycosylated Heavy Chain; Ph. Eur., European Pharmacopoeia; RH, Relative Humidity; SE-UPLC, Size Exclusion Ultra High Performance Liquid Chromatography; USP, United States Pharmacopoeia

[0322] Results of long-term stability test In the long-term stability investigation test, the combined DP was physically and chemically stable when stored at 5 °C throughout the evaluation period (Table 71). After 6 months at 5 °C, no perceptible changes in physical or chemical stability were detected in any of the monitored characteristics. The stability investigation test to verify the DP under long-term storage conditions is continued over 84 months.

[0323] Results of accelerated stability test The results of the stability investigation test from the analysis of the DP under the accelerated conditions of 25 °C / 60% RH are presented in Table 72. Over the 6-month evaluation period, an increase of 0.9% in HMW species and an increase of 0.3% in LMW species were observed by SE-UPLC. Decreases of 2.0%, 3.1%, and 5.1% were observed by CEX-UPLC at the major peaks corresponding to H1H17203P, H1H17139P, and H1H17161P, respectively. The DP maintained its potency after incubation at 25 °C / 60% RH for 6 months. The level of subvisible particles determined by HIAC was within the acceptance criteria range. No significant changes were observed in the other monitored characteristics. The test investigation to verify the DP under the accelerated storage conditions of 25 °C / 60% RH was completed.

[0324] Results of stress stability test The results of the stability investigation test from the analysis of DP under the stress conditions of 40°C / 75% RH are presented in Table 72. Over the 3-month evaluation period, an increase of 3.0% in HMW and 1.3% in LMW was observed by SE-UPLC. A decrease in purity of 3.8% or 7.8%, and an increase in LMW species of 1.8% or 6.4% were observed by reduced MCE or non-reduced MCE. Decreases of 9.3%, 11.5%, and 11.4% were observed by CEX-UPLC at the major peaks corresponding to H1H17203P, H1H17139P, and H1H17161P, respectively. The DP maintained its potency when determined by the pseudovirus neutralization assay. The DP maintained its potency after incubation at 40°C / 75% RH for 28 days, but showed a decrease in potency after 3 months of incubation when determined by the ADCC assay. The level of subvisible particles determined by HIAC was within the acceptance criteria range. No significant changes were observed in the other monitored characteristics. The test investigation to verify the DP under the stress storage conditions of 40°C / 75% RH was completed.

[0325] The results of the stability investigation from the analysis of DP after agitation and freeze / thaw conditions are presented in Table 73. The DP was physically and chemically stable when agitated for 120 minutes (vortexed at ambient temperature) or when subjected to 4 freeze / thaw cycles (-30°C freeze and room temperature thaw). No significant changes were observed in any of the monitored characteristics in the physical or chemical stability of the DP. The test investigation to verify the DP under agitation and freeze / thaw conditions was completed.

[0326] Conclusion on Stability The recommended storage for the combination DP is 2°C to 8°C. Data from the DP stability tests indicate that the formulation remains stable when stored at 2°C to 8°C.

[0327] (Table 71) Stability investigation of the 100 mg / mL co-formulated preparation of H1H17203P - H1H17139P - H1H17161P stored at 5°C TIFF2025090691000078.tif 193165 HMW, high molecular weight; LEFVP, liquid essentially free of visible particles; LMW, low molecular weight; MCE, microchip capillary electrophoresis; MFI, Micro-Flow Imaging (trademark); NGHC, non-glycosylated heavy chain; SE, size exclusion; UPLC, ultra-high performance liquid chromatography; CEX, cation exchange chromatography; RP, reverse phase; ADCC, antibody-dependent cell cytotoxicity activity; NR, not required

[0328] (Table 72) Stability investigation of 100 mg / mL H1H17203P-H1H17139P-H1H17161P co-formulated preparation stored at 25°C / 60% RH and 40°C / 75% RH TIFF2025090691000079.tif 210165 HMW, high molecular weight; LEFVP, liquid essentially free of visible particles; LMW, low molecular weight; MCE, microchip capillary electrophoresis; MFI, Micro-Flow Imaging (trademark); NGHC, non-glycosylated heavy chain; SE, size exclusion; UPLC, ultra-high performance liquid chromatography; CEX, cation exchange chromatography; ADCC, antibody-dependent cell cytotoxicity activity; NR, not required

[0329] (Table 73) Stability investigation of H1H17203P-H1H17139P-H1H17161P co-formulated preparation - effects of agitation and freeze-thaw TIFF2025090691000080.tif 215167 HMW, high molecular weight; LEFVP, liquid essentially free of visible particles; LMW, low molecular weight; MCE, microchip capillary electrophoresis; MFI, Micro-Flow Imaging (trademark); NGHC, non-glycosylated heavy chain; SE, size exclusion; UPLC, ultra-high performance liquid chromatography; CEX, cation exchange chromatography; ADCC, antibody-dependent cell cytotoxicity activity; NR, not required

[0330] Antibody sequence Table 74 describes the heavy chain variable region and light chain variable region of the selected anti-Ebola virus antibodies, as well as the amino acid sequence identifiers of the CDRs. Table 75 provides the sequence identifiers of the amino acid sequences of the full-length heavy chain and full-length light chain.

[0331] (Table 74) Amino acid sequence identifiers TIFF2025090691000081.tif27168

[0332] (Table 75) Sequence identifiers of the full-length heavy chain sequence and full-length light chain sequence TIFF2025090691000082.tif32128

[0333] The present disclosure is not limited to the specific embodiments described herein. Indeed, various modifications of the invention will become apparent to those skilled in the art from the foregoing description and the accompanying drawings in addition to those described herein. Such modifications are intended to be within the scope of the appended claims.

Claims

Claim 1. A stable liquid pharmaceutical preparation comprising: (a) sucrose at a concentration of 10% ± 2% w / v; (b) a buffer containing histidine at a concentration of less than 10 mM ± 2 mM; (c) an organic co-solvent containing polysorbate at a concentration of less than 0.2% ± 0.1% w / v; and (d) a first antibody comprising three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCV R / LCV R amino acid sequence pair of SEQ ID NO: 2 / 10; a second antibody comprising three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCV R / LCV R amino acid sequence pair of SEQ ID NO: 20 / 28; and a third antibody comprising three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained in the HCV R / LCV R amino acid sequence pair of SEQ ID NO: 38 / 46; wherein the concentration of each of the first, second, and third antibodies in the preparation is from 10 mg / mL ± 0.75 mg / mL to 40 mg / mL ± 7.5 mg / mL; the preparation has a pH of 6.0 ± 0.3; the pharmaceutical preparation. Claim 2. The pharmaceutical preparation according to claim 1, wherein the concentration of each of the first, second, and third antibodies is 16.67 mg / mL ± 1.25 mg / mL. Claim 3. The pharmaceutical preparation according to claim 1, wherein the concentration of each of the first, second, and third antibodies is 33.33 mg / mL ± 2.5 mg / mL. Claim 4. The pharmaceutical preparation according to any one of claims 1 to 3, further comprising L-histidine monohydrochloride monohydrate at a concentration of 25 mM ± 5 mM or less. Claim 5. The pharmaceutical preparation according to any one of claims 1 to 4, wherein the first antibody comprises HCDR1 of SEQ ID NO: 4, HCDR2 of SEQ ID NO: 6, HCDR3 of SEQ ID NO: 8, LCDR1 of SEQ ID NO: 12, LCDR2 of SEQ ID NO: 14, and LCDR3 of SEQ ID NO:

16. Claim 6. The pharmaceutical preparation according to claim 5, wherein the first antibody comprises HCV R of SEQ ID NO: 2 and LCV R of SEQ ID NO:

10. Claim 7. The pharmaceutical preparation according to claim 5, wherein the first antibody comprises the heavy chain of SEQ ID NO: 17 and the light chain of SEQ ID NO:

18.

8. The pharmaceutical preparation according to any one of claims 1 to 7, wherein the second antibody comprises HCDR1 of SEQ ID NO: 22, HCDR2 of SEQ ID NO: 24, HCDR3 of SEQ ID NO: 26, LCDR1 of SEQ ID NO: 30, LCDR2 of SEQ ID NO: 32, and LCDR3 of SEQ ID NO:

34.

9. The pharmaceutical preparation according to claim 8, wherein the second antibody comprises HCVR of SEQ ID NO: 20 and LCVR of SEQ ID NO:

28.

10. The pharmaceutical preparation according to claim 8, wherein the second antibody comprises a heavy chain of SEQ ID NO: 35 and a light chain of SEQ ID NO:

36.

11. The pharmaceutical preparation according to any one of claims 1 to 10, wherein the third antibody comprises HCDR1 of SEQ ID NO: 40, HCDR2 of SEQ ID NO: 42, HCDR3 of SEQ ID NO: 44, LCDR1 of SEQ ID NO: 48, LCDR2 of SEQ ID NO: 50, and LCDR3 of SEQ ID NO:

52.

12. The pharmaceutical preparation according to claim 11, wherein the third antibody comprises HCVR of SEQ ID NO: 38 and LCVR of SEQ ID NO:

46.

13. The pharmaceutical preparation according to claim 11, wherein the third antibody comprises a heavy chain of SEQ ID NO: 53 and a light chain of SEQ ID NO:

54.

14. The pharmaceutical preparation according to any one of claims 1 to 13, contained in a container.

15. The pharmaceutical preparation according to claim 14, wherein the container is a vial, and the total volume of the pharmaceutical preparation contained in each vial is 14.5 mL ± 0.5 mL.

16. The pharmaceutical preparation according to claim 15, wherein the pharmaceutical preparation contained in each vial comprises L-histidine at 0.74 mg / mL ± 0.05 mg / mL, L-histidine monohydrochloride monohydrate at 1.09 mg / mL ± 0.05 mg / mL, polysorbate 80 at 1 mg / mL ± 0.05 mg / mL, sucrose at 100 mg / mL ± 5 mg / mL, and water.

17. The pharmaceutical preparation according to any one of claims 1 to 16, formulated for intravenous administration.

18. The pharmaceutical preparation according to claim 17, which is diluted in a PVC intravenous (IV) bag containing 0.9% sodium chloride, 5% dextrose, or lactated Ringer's solution before intravenous administration.

19. The pharmaceutical preparation according to any one of claims 1 to 18, for administration to a subject at a dose of 150 mg / kg of the subject's body weight ± 15 mg / kg of the subject's body weight, comprising the first antibody, the second antibody, and the third antibody in a weight ratio of 1:1:

1.

20. The pharmaceutical formulation according to claim 19, wherein the final concentration of each of the diluted first antibody, second antibody, and third antibody is from about 2 mg / mL to about 30 mg / mL.

21. The pharmaceutical formulation according to claim 19 or claim 20, wherein the duration of intravenous administration of the diluted formulation is from about 2 hours to about 4 hours.

22. (a) A first antibody containing 16.67 mg / mL of HCDR1 of SEQ ID NO: 4, HCDR2 of SEQ ID NO: 6, HCDR3 of SEQ ID NO: 8, LCDR1 of SEQ ID NO: 12, LCDR2 of SEQ ID NO: 14, and LCDR3 of SEQ ID NO: 16, (b) A second antibody containing 16.67 mg / mL of HCDR1 of SEQ ID NO: 22, HCDR2 of SEQ ID NO: 24, HCDR3 of SEQ ID NO: 26, LCDR1 of SEQ ID NO: 30, LCDR2 of SEQ ID NO: 32, and LCDR3 of SEQ ID NO: 34, and (c) A third antibody containing 16.67 mg / mL of HCDR1 of SEQ ID NO: 40, HCDR2 of SEQ ID NO: 42, HCDR3 of SEQ ID NO: 44, LCDR1 of SEQ ID NO: 48, LCDR2 of SEQ ID NO: 50, and LCDR3 of SEQ ID NO: 52 contained in a volume of 14.5 mL, a stable liquid pharmaceutical formulation.

23. The first antibody contains HCVR of SEQ ID NO: 2 and LCVR of SEQ ID NO: 10, the second antibody contains HCVR of SEQ ID NO: 20 and LCVR of SEQ ID NO: 28, and the third antibody contains HCVR of SEQ ID NO: 38 and LCVR of SEQ ID NO: 46, the stable liquid pharmaceutical formulation according to claim 22.

24. The first antibody contains the heavy chain of SEQ ID NO: 17 and the light chain of SEQ ID NO: 18, the second antibody contains the heavy chain of SEQ ID NO: 35 and the light chain of SEQ ID NO: 36, and the third antibody contains the heavy chain of SEQ ID NO: 53 and the light chain of SEQ ID NO: 54, the stable liquid pharmaceutical formulation according to claim 22.

25. (d) 0.74 mg / mL of L-histidine, (e) 1.09 mg / mL of L-histidine monohydrochloride monohydrate, (f) 1 mg / mL of polysorbate 80, (g) 100 mg / mL of sucrose, and (h) water further contained, and the formulation has a pH of 6.0, the stable liquid pharmaceutical formulation according to any one of claims 22 to 24. **Claim 26** (a) A first antibody containing 33.33 mg / mL of HCDR1 of SEQ ID NO: 4, HCDR2 of SEQ ID NO: 6, HCDR3 of SEQ ID NO: 8, LCDR1 of SEQ ID NO: 12, LCDR2 of SEQ ID NO: 14, and LCDR3 of SEQ ID NO: 16, (b) A second antibody containing 33.33 mg / mL of HCDR1 of SEQ ID NO: 22, HCDR2 of SEQ ID NO: 24, HCDR3 of SEQ ID NO: 26, LCDR1 of SEQ ID NO: 30, LCDR2 of SEQ ID NO: 32, and LCDR3 of SEQ ID NO: 34, and (c) A third antibody containing 33.33 mg / mL of HCDR1 of SEQ ID NO: 40, HCDR2 of SEQ ID NO: 42, HCDR3 of SEQ ID NO: 44, LCDR1 of SEQ ID NO: 48, LCDR2 of SEQ ID NO: 50, and LCDR3 of SEQ ID NO: 52 in a volume of 14.5 mL, a stable liquid pharmaceutical preparation. **Claim 27** The first antibody contains HCV R of SEQ ID NO: 2 and LCVR of SEQ ID NO: 10, the second antibody contains HCV R of SEQ ID NO: 20 and LCVR of SEQ ID NO: 28, and the third antibody contains HCV R of SEQ ID NO: 38 and LCVR of SEQ ID NO: 46, The stable liquid pharmaceutical preparation according to Claim 26. **Claim 28** The first antibody contains a heavy chain of SEQ ID NO: 17 and a light chain of SEQ ID NO: 18, the second antibody contains a heavy chain of SEQ ID NO: 35 and a light chain of SEQ ID NO: 36, and the third antibody contains a heavy chain of SEQ ID NO: 53 and a light chain of SEQ ID NO: 54, The stable liquid pharmaceutical preparation according to Claim 26. **Claim 29** (d) 0.74 mg / mL of L-histidine, (e) 1.09 mg / mL of L-histidine monohydrochloride monohydrate, (f) 1 mg / mL of polysorbate 80, (g) 100 mg / mL of sucrose, and (h) water further included, and the preparation has a pH of 6.0, The stable liquid pharmaceutical preparation according to any one of Claims 26 to 28.

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