Stable Liquid Pharmaceutical Formulation and Kit

Abstract

The present invention provides stable pharmaceutical formulations comprising a human antibody that specifically binds to human programmed death protein-1 (PD-1). In certain embodiments, the formulations contain, in addition to an anti-PD-1 antibody, a buffer, an amino acid, a nonionic surfactant, and a sugar. The pharmaceutical formulations of the present invention exhibit a substantial degree of antibody stability after stress and storage.

Description

1 / 130 Descriptive Report of the Invention Patent for a STABLE LIQUID PHARMACEUTICAL FORMULATION AND KIT.

[0001] The present application is being filed on March 23, 2018 as a PCT International Patent Application and claims the benefit of priority to Provisional Patent Application No. US 62 / 482,270, filed on April 6, 2017, the description of which is incorporated herein by reference in full. FIELD OF THE INVENTION

[0002] The present invention relates to the field of therapeutic antibody formulations. More specifically, the present invention relates to the field of pharmaceutical formulations comprising a human antibody that specifically binds to human programmed death protein-1 (PD-1). BACKGROUND OF THE INVENTION

[0003] Therapeutic macromolecules (e.g., antibodies) must be formulated in a way that not only makes the molecules suitable for administration to patients but also maintains their stability during storage and subsequent use. For example, therapeutic antibodies in liquid solution are prone to degradation, aggregation, or undesirable chemical modifications unless the solution is properly formulated. The stability of an antibody in a liquid formulation depends not only on the types of excipients used in the formulation but also on the amounts and proportions of the excipients relative to each other. Furthermore, considerations beyond 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 antibody concentration that can be accommodated by a given formulation, and the visual quality or appeal of the formulation. Thus, when formulating a Petition 870220073975, dated 08 / 17 / 2022, page 12 / 245 2 / 130 therapeutic antibody, great care must be taken to arrive at a formulation that remains stable, contains an adequate concentration of antibodies, and has an appropriate viscosity, in addition to other properties that allow the formulation to be conveniently administered to patients.

[0004] Antibodies against human programmed death protein-1 (PD-1) are an example of a therapeutically relevant macromolecule that requires appropriate formulation. Anti-PD-1 antibodies are clinically useful for the treatment of cancer (e.g., lung cancer, melanoma, and brain cancer) and viral infections and autoimmune diseases. Exemplary anti-PD-1 antibodies are described, inter alia, in US7101550, US7595048, US7488802, US7563869, US8008449, US8168757, US8216996, US20110008369, US20130017199, US 20130022595, and in documents WO2006121168, WO2009114455, WO2009114453, WO2009101611, EP2262837, and EP2504028. Document US20140234296 describes lyophilized formulations of an anti-PD-1 antibody.

[0005] Although anti-PD-1 antibodies are known, there is still a need in the art for innovative pharmaceutical formulations comprising anti-PD-1 antibodies that are sufficiently stable and suitable for administration to patients. BRIEF SUMMARY OF THE INVENTION

[0006] The present invention satisfies the aforementioned need by providing stable pharmaceutical formulations comprising a human antibody that specifically binds to human programmed death protein-1 (PD-1).

[0007] In one aspect, a stable, low-viscosity liquid pharmaceutical formulation is provided comprising: (i) a human antibody that specifically binds to human programmed death protein-1 (PD-1); (ii) a buffer; (iii) an organic cosolvent; (iv) a Petition 870220073975, dated 08 / 17 / 2022, page 13 / 245 3 / 130 stabilizer; and (v) a viscosity modifier.

[0008] In several embodiments, the antibody is provided at a concentration from about 5 ± 0.75 mg / mL to about 250 ± 37.5 mg / mL. In one embodiment, the antibody is provided at a concentration of 12.5 mg / mL ± 1.85 mg / mL or about 12.5 mg / mL. In one embodiment, the antibody is provided at a concentration of 25 mg / mL ± 3.75 mg / mL or about 25 mg / mL. In another embodiment, the antibody is provided at a concentration of 50 mg / mL ± 7.5 mg / mL or about 50 mg / mL. In another embodiment, the antibody is provided at a concentration of 100 mg / mL ± 15 mg / mL or about 100 mg / mL. In one embodiment, the antibody is provided at a concentration of 150 mg / mL ± 22.5 mg / mL or approximately 150 mg / mL. In another embodiment, the antibody is provided at a concentration of 175 mg / mL ± 26.25 mg / mL or approximately 175 mg / mL. In yet another embodiment, the antibody is provided at a concentration of 200 mg / mL ± 30 mg / mL or approximately 200 mg / mL.

[0009] In certain embodiments, the formulation comprises any of the anti-PD-1 antibodies described in US20150203579, incorporated herein in full. In certain embodiments, the anti-PD-1 antibody comprises (a) a heavy chain variable region (HCVR) comprising the heavy chain complementarity-determining regions 1, 2, and 3 (HCDR1-HCDR2-HCDR3), each comprising a sequence of SEQ ID NO: 3, SEQ ID NO: 4, and SEQ ID NO: 5, respectively; and (b) a light chain variable region (LCVR) comprising the light chain complementarity-determining regions 1, 2, and 3 (LCDR1-LCDR2-LCDR3), each comprising a sequence of SEQ ID NO: 6, SEQ ID NO: 7, and SEQ ID NO: 8, respectively. In one embodiment, the antibody comprises an HCVR comprising the amino acid sequence SEQ ID NO: 1 and an LCVR that Petition 870220073975, dated 08 / 17 / 2022, page 14 / 245 4 / 130 comprises the amino acid sequence of SEQ ID NO: 2. In one embodiment, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 9 and a light chain comprising the amino acid sequence of SEQ ID NO: 10. In one embodiment, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 11 and a light chain comprising the amino acid sequence of SEQ ID NO: 10. In one embodiment, the antibody comprises a heavy chain comprising the amino acid sequence selected from the group consisting of SEQ ID NOs: 9 and 11; and a light chain comprising the amino acid sequence of SEQ ID NO: 10. In one embodiment, the antibody comprises an HCVR that has 90% sequence identity with SEQ ID NO: 1. In one embodiment, the antibody comprises an LCVR that has 90% sequence identity with SEQ ID NO: 2.In one embodiment, the antibody comprises an HCVR that has 90% sequence identity with SEQ ID NO: 1 and an LCVR that has 90% sequence identity with SEQ ID NO: 2.

[0010] 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 approximately 6.0 ± 0.3.

[0011] In one embodiment, the buffer comprises histidine. In certain embodiments, the histidine buffer is at a concentration from 5 mM ± 1 mM to 50 mM ± 10 mM, preferably from 5 mM ± 1 mM to 25 mM ± 5 mM. In one embodiment, the histidine buffer is at a concentration of 10 mM ± 2 mM or about 10 mM. In one embodiment, the histidine buffer is at a concentration of 20 mM ± 4 mM or about 20 mM. In one embodiment, the histidine buffer is at a concentration of 40 nM. Petition 870220073975, dated 08 / 17 / 2022, page 15 / 245 5 / 130 ± 8 mM or approximately 40 nM. In certain embodiments, the histidine buffer comprises L-histidine and L-histidine monohydrochloride monohydrate. In one embodiment, L-histidine is at a concentration from 2 mM ± 0.4 mM to 25 mM ± 5 mM, preferably from 4 mM ± 0.8 mM to 20 mM ± 4 mM. In another embodiment, L-histidine monohydrochloride monohydrate is at a concentration from 2 mM ± 0.4 mM to 25 mM ± 5 mM, preferably from 4 mM ± 0.8 mM to 20 mM ± 4 mM. In one embodiment, the 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. In another embodiment, the buffer comprises histidine at a concentration of 10 mM ± 2 mM, wherein the histidine 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.

[0012] In certain embodiments, the organic cosolvent is a nonionic polymer containing a polyoxyethylene moiety. In one embodiment, the organic solvent is a surfactant. In some embodiments, the organic cosolvent is any one or more of polysorbate, poloxamer 188, and polyethylene glycol 3350. In one embodiment, the organic cosolvent is polysorbate 80. In one embodiment, the organic cosolvent is polysorbate 20.

[0013] In one embodiment, the organic cosolvent is at a concentration from about 0.01% ± 0.005% to about 1% ± 0.5% by weight by volume or w / v, where, for example, 0.1 g / ml = 10% and 0.01 g / ml = 1%. In certain embodiments, the organic solvent is polysorbate at a concentration from 0.05% ± 0.025% to 0.5% ± 0.25% (w / v). In one embodiment, the organic cosolvent is polysorbate 80, which is at a concentration of 0.2% ± 0.1% w / v or about 0.2%. In another embodiment, the organic cosolvent is Petition 870220073975, dated 08 / 17 / 2022, page 16 / 245 6 / 130 polysorbate 80, which is at a concentration of 0.1% ± 0.05% w / v or about 0.1% w / v. In one embodiment, the organic cosolvent is polysorbate 20, which is at a concentration of 0.2% ± 0.1% w / v or about 0.2%. In another embodiment, the organic cosolvent is polysorbate 20, which is at a concentration of 0.1% ± 0.05% w / v or about 0.1% w / v.

[0014] In certain embodiments, the stabilizer is a sugar. In one embodiment, the sugar is sucrose. In several embodiments, the stabilizer is at a concentration from 1% ± 0.2% w / v to 20% ± 4% w / v, from 5% ± 1% w / v to 15% ± 3% w / v, or from 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 approximately 10% w / v.

[0015] In one embodiment, the viscosity modifier is an amino acid. In one embodiment, the viscosity modifier is L-proline. In certain embodiments, the viscosity modifier is at a concentration from 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%.

[0016] In certain embodiments, the viscosity of the liquid pharmaceutical formulation at 25 °C is less than or equal to about 15 cPoise ± 10%. In certain embodiments, the viscosity at 25 °C is between 1.0 cPoise ± 10% and 20 cPoise ± 10%. In certain embodiments, the viscosity of the liquid pharmaceutical formulation is < 15 cPoise. In certain embodiments, the viscosity of the formulation Petition 870220073975, dated 08 / 17 / 2022, p. 17 / 245 7 / 130 liquid pharmaceutical formulation has a viscosity < 20 cPoise. In certain embodiments, the viscosity of the liquid pharmaceutical formulation is <10 cPoise. In certain embodiments, the viscosity at 25 °C is 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%.

[0017] In one aspect, a stable liquid pharmaceutical formulation of low viscosity is provided comprising: (i) from 5 ± 0.75 mg / mL to 250 ± 37.5 mg / mL of a human antibody that specifically binds to human PD-1; (ii) histidine buffer from 0 mM to 40 ± 8 mM; (iii) polysorbate 80 from 0% to 0.5% ± 0.25% (w / v); (iv) sucrose from 0% to 15% ± 3% (w / v); and (v) proline from 0% to 5% ± 1% at a pH of about 5.3 to about 6.7; wherein the anti-PD-1 antibody comprises a heavy chain variable region (HCVR) and a light chain variable region (LCVR), such that the HCVR / LCVR combination comprises heavy and light chain complementarity-determining regions (HCDR1-HCDR2-HCDR3 / LCDR1-LCDR2-LCDR3) comprising the amino acid sequences SEQ ID NOs: 3-4-5 / SEQ ID NOs: 67-8, respectively.In one embodiment, the anti-PD-1 antibody comprises a heavy chain variable region (HCVR) and a light chain variable region (LCVR) comprising an amino acid sequence of SEQ ID NO: 1 and SEQ ID NO: 2, respectively. In certain embodiments, the anti-PD1 antibody comprises an Fc region selected from the group consisting of human IgG1, IgG2, IgG3, and IgG4 isotypes. In one embodiment, the antibody comprises a human IgG4 isotype. In one embodiment, the... Petition 870220073975, dated 08 / 17 / 2022, p. 18 / 245 The antibody comprises a heavy chain comprising the amino acid sequence selected from the group consisting of SEQ ID NOs: 9 and 11; and a light chain comprising the amino acid sequence of SEQ ID NO: 10. In one embodiment, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 9 and a light chain comprising the amino acid sequence of SEQ ID NO: 10. In another embodiment, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 11 and a light chain comprising the amino acid sequence of SEQ ID NO: 10. In one embodiment, the antibody has a molecular weight of 143 kDa ± 5 kDa.

[0018] In certain embodiments, a stable, low-viscosity liquid pharmaceutical formulation is provided comprising: (i) 5 ± 0.75 mg / mL to 250 ± 37.5 mg / mL of a human antibody that specifically binds to human PD-1; (ii) histidine buffer from 0 mM to 40 ± 8 mM; (iii) polysorbate 80 from 0% to 0.5% ± 0.25% (w / v); (iv) sucrose from 0% to 15% ± 3% (w / v); and (v) proline from 0% to 5% ± 1% at a pH of about 5.3 to about 6.7; wherein the anti-PD-1 antibody comprises an HCVR and an LCVR, wherein the HCVR has 90% sequence identity with SEQ ID NO: 1 and / or the LCVR has 90% sequence identity with SEQ ID NO: 2. In one embodiment, the anti-PD-1 antibody comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 1 and an LCVR comprising the amino acid sequence of SEQ ID NO: 2.In one embodiment, the anti-PD-1 antibody comprises a heavy chain comprising the amino acid sequence selected from the group consisting of SEQ ID NOs: 9 and 11; and a light chain comprising the amino acid sequence of SEQ ID NO: 10. Petition 870220073975, dated 08 / 17 / 2022, p. 19 / 245 9 / 130

[0019] In certain embodiments, a stable, low-viscosity liquid pharmaceutical formulation is provided comprising: (i) 5 ± 0.75 mg / mL to 250 ± 37.5 mg / mL of a human antibody that specifically binds to human PD-1; (ii) histidine buffer from 0 mM to 40 ± 8 mM; (iii) polysorbate 80 from 0% to 0.5% ± 0.25% (w / v); (iv) sucrose from 0% to 15% ± 3% (w / v); and (v) proline from 0% to 5% ± 1% at a pH of about 5.3 to about 6.7; wherein the anti-PD-1 antibody comprises an HCVR and an LCVR, wherein HCVR comprises an amino acid sequence of SEQ ID NO: 1 having no more than five amino acid substitutions and wherein LCVR comprises an amino acid sequence of SEQ ID NO: 2 having no more than two amino acid substitutions.In one embodiment, the anti-PD-1 antibody comprises a HCVR comprising the amino acid sequence of SEQ ID NO: 1 and a LCVR comprising the amino acid sequence of SEQ ID NO: 2. In another embodiment, the anti-PD-1 antibody comprises a heavy chain comprising the amino acid sequence selected from the group consisting of SEQ ID NOs: 9 and 11; and a light chain comprising the amino acid sequence of SEQ ID NO: 10.

[0020] In certain embodiments, the formulation of any of the preceding aspects has an attribute selected from the group consisting of: (i) the formulation is stable to long-term storage at 25°C, 5°C, -20°C, -30°C and -80°C, as described herein; (ii) the formulation is stable to agitation stress as described herein; (iii) the formulation is of low viscosity (viscosity less than 20 cPoise, preferably less than 15 cPoise); (iv) the formulation is stable even with variations of up to ± 50% in the excipient concentrations of the formulation, as described herein; (v) the formulation is iso-osmolar under physiological conditions; (vi) the formulation Petition 870220073975, dated 08 / 17 / 2022, page 20 / 245 10 / 130 is stable and compatible with intravenous administration devices and procedures; and (vi) the formulation is stable upon long-term storage in a glass vial or pre-filled syringe.

[0021] In certain embodiments of this aspect, a stable liquid formulation is provided comprising: (i) 5 ± 0.75 mg / mL to 250 ± 37.5 mg / mL of a human antibody that binds specifically to human PD-1; (ii) 5 mM ± 1 mM to 20 ± 4 mM of histidine buffer; (iii) polysorbate 80 from 0.05% ± 0.025% to 0.3% ± 0.15% (w / v); (iv) sucrose from 1% ± 0.2% to 10% ± 2% (w / v); and (v) proline from 1% ± 0.2% to 5% ± 1% at a pH of about 6.0, wherein the antibody comprises an HCVR / LCVR comprising a pair of amino acid sequences of SEQ ID Nos: 1 / 2. In one embodiment, the stable liquid formulation of this aspect has a viscosity less than 15 cP. In one embodiment, >90% of the antibodies have a molecular weight of 143 kDa ± 1 kDa. In one embodiment, the pharmaceutical formulation has a viscosity less than 20 cP, less than 15 cP, or less than 10 cP.In one embodiment, more than 96% of the antibodies have a native conformation when stored for 12 months at 5°C. In another embodiment, at least 97% or more of the antibodies have a native conformation when stored at -80°C, -30°C, and / or -20°C for 6 months.

[0022] In one embodiment of this aspect, the stable liquid formulation comprises (i) 25 ± 3.75 mg / mL of an anti-PD-1 antibody; (ii) histidine buffer at 10 ± 2 mM; (iii) polysorbate 80 at 0.2% ± 0.1% (w / v); (iv) proline at 1.5% ± 0.3% (w / v); and (v) sucrose at 5% ± 1% (w / v) at a pH of 6.0 ± 0.3, wherein the antibody comprises an HCVR / LCVR comprising a pair of amino acid sequences of SEQ ID NOs: 1 / 2. Petition 870220073975, dated 08 / 17 / 2022, p. 21 / 245 11 / 130

[0023] In one embodiment of this aspect, the stable liquid formulation comprises (i) 25 ± 3.75 mg / mL of an anti-PD-1 antibody; (ii) L-histidine at 4.8 mM ± 0.96 mM; (iii) L-histidine monohydrochloride monohydrate at 5.2 mM ± 1.04 mM; (iv) polysorbate 80 at 0.2% ± 0.1% (w / v); (v) proline at 1.5% ± 0.3% (w / v); and (vi) sucrose at 5% ± 1% (w / v) at a pH of 6.0 ± 0.3, wherein the antibody comprises an HCVR / LCVR comprising a pair of amino acid sequences of SEQ ID NOs: 1 / 2.

[0024] In one embodiment of this aspect, the stable liquid formulation comprises (i) 50 ± 7.5 mg / mL of an anti-DP-1 antibody; (ii) 10 ± 2 mM histidine buffer; (iii) 0.2% ± 0.1% (w / v) polysorbate 80; (iv) 1.5% ± 0.3% (w / v) proline; and (v) 5% ± 1% (w / v) sucrose at a pH of 6.0 ± 0.3, wherein the antibody comprises an HCVR / LCVR comprising a pair of amino acid sequences of SEQ ID NOs: 1 / 2. In one embodiment of this particular formulation, the viscosity is less than 10 cPoise.

[0025] In one embodiment of this aspect, the stable liquid formulation comprises (i) 50 ± 7.5 mg / mL of an anti-PD-1 antibody; (ii) L-histidine at 4.8 mM ± 0.96 mM; (iii) L-histidine monohydrochloride monohydrate at 5.2 mM ± 1.04 mM; (iv) polysorbate 80 at 0.2% ± 0.1% (w / v); (v) proline at 1.5% ± 0.3% (w / v); and (vi) sucrose at 5% ± 1% (w / v) at a pH of 6.0 ± 0.3, wherein the antibody comprises an HCVR / LCVR comprising a pair of amino acid sequences of SEQ ID NOs: 1 / 2.

[0026] In one embodiment, the stable liquid formulation comprises (i) 100 ± 15 mg / mL of an anti-PD-1 antibody; (ii) 10 ± 2 mM histidine buffer; (iii) 0.2% ± 0.1% (w / v) polysorbate 80; (iv) 1.5% ± 0.3% (w / v) proline; and (v) 5% ± 1% (w / v) sucrose at a pH of 6.0 ± 0.3, wherein the antibody comprises an HCVR / LCVR comprising a pair of amino acid sequences of SEQ ID Petition 870220073975, dated 08 / 17 / 2022, p. 22 / 245 12 / 130 NOs: 1 / 2. In one embodiment of this particular formulation, the viscosity is less than 10 cPoise.

[0027] In one embodiment of this aspect, the stable liquid formulation comprises (i) 100 ± 15 mg / mL of an anti-PD-1 antibody; (ii) L-histidine at 4.8 mM ± 0.96 mM; (iii) L-histidine monohydrochloride monohydrate at 5.2 mM ± 1.04 mM; (iv) polysorbate 80 at 0.2% ± 0.1% (w / v); (v) proline at 1.5% ± 0.3% (w / v); and (vi) sucrose at 5% ± 1% (w / v) at a pH of 6.0 ± 0.3, wherein the antibody comprises an HCVR / LCVR comprising a pair of amino acid sequences of SEQ ID NOs: 1 / 2.

[0028] In one embodiment, the stable liquid formulation comprises (i) 150 ± 22.5 mg / mL of an anti-PD-1 antibody; (ii) 10 ± 2 mM histidine buffer; (iii) 0.2% ± 0.1% (w / v) polysorbate 80; (iv) 10% ± 2% (w / v) sucrose; and (v) 1.5% ± 0.3% (w / v) proline at a pH of 6.0 ± 0.3, wherein the antibody comprises an HCVR / LCVR comprising a pair of amino acid sequences of SEQ ID NOs: 1 / 2. In one embodiment of this particular formulation, the viscosity is less than 20 cPoise, preferably less than 15 cPoise.

[0029] In one embodiment of this aspect, the stable liquid formulation comprises (i) 150 ± 22.5 mg / mL of an anti-PD-1 antibody; (ii) L-histidine at 4.8 mM ± 0.96 mM; (iii) L-histidine monohydrochloride monohydrate at 5.2 mM ± 1.04 mM; (iv) polysorbate 80 at 0.2% ± 0.1% (w / v); (v) proline at 1.5% ± 0.3% (w / v); and (vi) sucrose at 5% ± 1% (w / v) at a pH of 6.0 ± 0.3, wherein the antibody comprises an HCVR / LCVR comprising a pair of amino acid sequences of SEQ ID NOs: 1 / 2.

[0030] In one embodiment of this aspect, the stable liquid formulation comprises (i) 175 ± 26.25 mg / mL of an anti-PD-1 antibody; (ii) 10 ± 2 mM histidine buffer; (iii) 0.2% ± 0.1% (w / v) polysorbate 80; (iv) 5% ± 1% (w / v) sucrose; and (v) 1.5% ± 0.3% proline. Petition 870220073975, dated 08 / 17 / 2022, p. 23 / 245 13 / 130 (w / v) at a pH of 6.0 ± 0.3, wherein the antibody comprises an HCVR / LCVR comprising a pair of amino acid sequences of SEQ ID NOs: 1 / 2. In one embodiment of this particular formulation, the viscosity is less than 20 cPoise, preferably less than 15 cPoise.

[0031] In one embodiment of this aspect, the stable liquid formulation comprises (i) 175 ± 26.25 mg / mL of an anti-PD-1 antibody; (ii) L-histidine at 4.8 mM ± 0.96 mM; (iii) L-histidine monohydrochloride monohydrate at 5.2 mM ± 1.04 mM; (iv) polysorbate 80 at 0.2% ± 0.1% (w / v); (v) proline at 1.5% ± 0.3% (w / v); and (vi) sucrose at 5% ± 1% (w / v) at a pH of 6.0 ± 0.3, wherein the antibody comprises an HCVR / LCVR comprising a pair of amino acid sequences of SEQ ID NOs: 1 / 2.

[0032] In one embodiment of this aspect, the stable liquid formulation comprises (i) 200 ± 30.00 mg / mL of an anti-PD-1 antibody; (ii) 10 ± 2 mM histidine buffer; (iii) 0.2% ± 0.1% (w / v) polysorbate 80; (iv) 5% ± 1% (w / v) sucrose; and (v) 1.5% ± 0.3% (w / v) proline at a pH of 6.0 ± 0.3, wherein the antibody comprises an HCVR / LCVR comprising a pair of amino acid sequences of SEQ ID NOs: 1 / 2. In one embodiment of this particular formulation, the viscosity is less than 20 cPoise.

[0033] In one embodiment of this aspect, the stable liquid formulation comprises (i) 200 ± 30.00 mg / mL of an anti-DP-1 antibody; (ii) L-histidine at 4.8 mM ± 0.96 mM; (iii) L-histidine monohydrochloride monohydrate at 5.2 mM ± 1.04 mM; (iv) polysorbate 80 at 0.2% ± 0.1% (w / v); (v) proline at 1.5% ± 0.3% (w / v); and (vi) sucrose at 5% ± 1% (w / v) at a pH of 6.0 ± 0.3, wherein the antibody comprises an HCVR / LCVR comprising a pair of amino acid sequences of SEQ ID NOs: 1 / 2.

[0034] In one embodiment, after storage of the formulation a Petition 870220073975, dated 08 / 17 / 2022, page 24 / 245 14 / 130 At 45°C for 28 days, >90% of the antibody is native and >35% of the antibody is in the form of a primary charge. In one embodiment, after storing the formulation at 25°C for three months, >94% of the antibody is native and >44% of the antibody is in the form of a primary charge. In one embodiment, after storing the formulation at 5°C for 12 months, >96% of the antibody is native and >50% of the antibody is in the form of a primary charge. In one embodiment, after storing the formulation at 20°C for 12 months, >96% of the antibody is native and >40% of the antibody is in the form of a primary charge. In one embodiment, after storing the formulation at 30°C for 12 months, >96% of the antibody is native and >40% of the antibody is in the form of a primary charge. In one embodiment, after storing the formulation at -80°C for 12 months, > 96% of the antibody is native and > 40% of the antibody is in the form of a primary charge.In one embodiment, more than 96% of the antibodies have a native conformation when stored for 12 months at 5°C. In another embodiment, at least 97% or more of the antibodies have a native conformation when stored at -80°C, -30°C, and / or -20°C for 6 months.

[0035] In one aspect, the present invention provides a stable liquid formulation comprising: (i) up to 100 mg / mL of an anti-PD-1 antibody; (ii) histidine buffer from 2 mM ± 0.4 mM to 20 mM ± 4 mM; (iii) sucrose up to 20% ± 4% (w / v); and (iv) polysorbate up to 0.2% ± 0.1% w / v at a pH of 6.0 ± 0.3. In one embodiment, the stable liquid formulation comprises 25 mg / mL of anti-PD-1 antibody. In one embodiment, the stable liquid formulation comprises 50 mg / mL of anti-PD-1 antibody. In one embodiment, the stable liquid formulation comprises 75 mg / mL of anti-PD-1 antibody. In one embodiment, the stable liquid formulation comprises histidine buffer at 10 mM ± 2 mM. In one modality, the formulation Petition 870220073975, dated 08 / 17 / 2022, page 25 / 245 15 / 130 stable liquid comprises 5% sucrose. In one embodiment, the stable liquid formulation comprises 6% sucrose. In one embodiment, the stable liquid formulation comprises 9% sucrose. In one embodiment, the stable liquid formulation comprises 10% sucrose. In one embodiment, the stable liquid formulation comprises 0.1% polysorbate. In one embodiment, the polysorbate is polysorbate 80 or polysorbate 20. In one embodiment, the anti-PD-1 antibody comprises an HCVR / LCVR of SEQ ID NOs: 1 / 2.

[0036] In one aspect, a stable liquid pharmaceutical formulation of any of the preceding aspects is provided in a container. In one embodiment, the container is a polycarbonate bottle. In one embodiment, the container is a glass bottle. In one embodiment, the glass bottle is a type 1 borosilicate glass bottle with a fluorocarbon-coated butyl rubber stopper. In one embodiment, the container is a microinfuser. In one embodiment, the container is a syringe. In one embodiment, the container is a pre-filled syringe. In one embodiment, the syringe comprises a fluorocarbon-coated plunger. In certain embodiments, the syringe is a 1 ml or 2.25 ml glass syringe containing less than about 500 parts per billion of tungsten, fitted with a 27-G needle, a fluorocarbon-coated butyl rubber stopper, and a non-cytotoxic, latex-free rubber needle guard.In one embodiment, the syringe is a 1 mL glass syringe fitted with a 27 G thin-walled needle, a FLUROTEC-coated 4023 / 50 rubber stopper, and an FM 27 rubber needle protector. In another embodiment, the syringe is a 1 mL, 2 mL, 3 mL, 5 mL, or 10 mL plastic syringe fitted with a needle.

[0037] In one aspect, a kit is provided comprising a stable pharmaceutical composition of any of the foregoing aspects, a container and instructions. In one embodiment, the container Petition 870220073975, dated 08 / 17 / 2022, p. 26 / 245 16 / 130 is a glass vial. In one embodiment, the container is a pre-filled syringe. In one embodiment, the syringe is a 1 mL or 2.25 mL glass syringe fitted with a 27 G thin-walled needle, a FLUROTEC-coated 4023 / 50 rubber stopper, and an FM 27 rubber needle protector. In one embodiment, the syringe is a 1 mL, 2 mL, 3 mL, 5 mL, or 10 mL plastic syringe fitted with a needle.

[0038] In certain embodiments, the present invention provides a pre-filled syringe comprising a stable liquid pharmaceutical formulation comprising: (i) from 5 ± 0.75 mg / mL to 250 ± 37.5 mg / mL of a human antibody that specifically binds to human PD 1; (ii) histidine buffer from 5 mM ± 1 mM to 20 ± 4 mM; (iii) polysorbate 80 from 0.05% ± 0.025% to 0.3% ± 0.15% (w / v); (iv) sucrose from 1% ± 0.2% to 10% ± 2% (w / v); and (v) proline from 1% ± 0.2% to 5% ± 1% at a pH of 6.0 ± 0.3, wherein the antibody comprises an HCVR / LCVR comprising a pair of amino acid sequences of SEQ ID NOs: 1 / 2; wherein the formulation has an attribute selected from the group consisting of: (i) 98% of the antibody is in native form after storage at 5°C for 12 months; (ii) > 53% of the antibody is the mainload variant after storage at 5°C for 12 months;(iii) > 97% of the antibody is in its native form after storage at 25°C for 6 months; (iv) the formulation is stable to shake stress, where > 98% of the antibody is in its native form after 120 minutes of shake stress in the pre-filled syringe; (v) more than 90% of the antibodies have a molecular weight of 143 kDa ± 1 kDa; (vi) the pharmaceutical formulation has a viscosity less than 20 cP, less than 15 cP, or less than 10 cP; (vii) more than 96% of the antibodies have a native conformation when stored for 12 months at 5°C; and (viii) at least 97% or more of the antibodies have a; Petition 870220073975, dated 08 / 17 / 2022, p. 27 / 245 17 / 130 native conformation when stored at -80°C, -30°C and / or 20°C for 6 months.

[0039] In certain embodiments, the present invention provides a glass vial comprising a stable liquid pharmaceutical formulation comprising: (i) from 5 ± 0.75 mg / mL to 250 ± 37.5 mg / mL of a human antibody that specifically binds to human PD1; (ii) histidine buffer from 5 mM ± 1 mM to 20 ± 4 mM; (iii) polysorbate 80 from 0.05% ± 0.025% to 0.3% ± 0.15% (w / v); (iv) sucrose from 1% ± 0.2% to 10% ± 2% (w / v); and (v) proline from 1% ± 0.2% to 5% ± 1% at a pH of 6.0 ± 0.3, wherein the antibody comprises an HCVR / LCVR comprising a pair of amino acid sequences with SEQ ID Nos: 1 / 2; wherein the formulation has an attribute selected from the group consisting of: (i) the formulation is stable to storage and stress in a glass vial; (ii) the formulation is stable and compatible for use in IV administration devices;(iii) the formulation is chemically and physically stable to dilution with standard diluents known in the art (e.g., 0.9% sodium chloride or 5% dextrose); (iv) the formulation is stable in IV bags made of glass or polymeric plastics (e.g., polyvinyl chloride, phthalates, polyolefins, or polypropylene); (v) the formulation is compatible with standard infusion pumps (e.g., peristaltic pump, fluid displacement pump); (vi) > 90% of the antibodies have a molecular weight of 143 kDa ± 1 kDa; (vii) the pharmaceutical formulation has a viscosity less than 20 cP, less than 15 cP, or less than 10 cP; (viii) more than 96% of the antibodies have a native conformation when stored for 12 months at 5°C; and (ix) at least 97% or more of the antibodies have a native conformation when stored at -80°C, -30°C and / or -20°C for 6 months.;

[0040] Other modalities will become evident from a Petition 870220073975, dated 08 / 17 / 2022, p. 28 / 245 18 / 130 detailed description review below. BRIEF DESCRIPTION OF THE FIGURES

[0041] Figure 1 is a table showing the effect of pH on the stability of 150 mg / mL of mAb1 incubated at 45°C for 28 days.aThe SE-UPLC and CEX-UPLC results for 'Raw Material' are the average values ​​of the Raw Material for all formulations.

[0042] Figure 2 shows the storage stability of three formulations F1, F2, and F3, where F1 comprises 210 mg / mL of mAb1, 10 mM histidine, and 3% proline at pH 6.0; F2 comprises 210 mg / mL of mAb1, 10 mM histidine, and 3% sucrose at pH 6.0; and F3 comprises 210 mg / mL of mAb1, 10 mM histidine, and 5% sucrose at pH 6.0. Storage stability is measured by the % of high molecular weight (HMW) species generated when stored at -80°C (A), -30°C (B), and -20°C (C) for up to 9 months and analyzed by size exclusion chromatography (SEC).

[0043] Figure 3 shows the storage stability of three formulations F1, F2 and F3, wherein F1 comprises 150 mg / mL of mAb1, 10 mM histidine, 9% sucrose and 0.2% polysorbate 80 (PS80) at pH 6.0; F2 comprises 175 mg / mL of mAb1, 10 mM histidine, 3% proline and 0.2% PS80 at pH 6.0; and F3 comprises 175 mg / mL of mAb1, 10 mM histidine, 5% sucrose, 1.5% proline and 0.2% PS80 at pH 6.0. Storage stability is measured by the % of high molecular weight (HMW) species generated when stored at -80°C (A), -30°C (B) and -20°C (C) for up to 6 months and analyzed by size exclusion chromatography (SEC).

[0044] Figure 4 is a table showing the viscosity of 150 mg / mL mAb1 with the addition of excipients and viscosity modifiers. Petition 870220073975, dated 08 / 17 / 2022, page 29 / 245 19 / 130

[0045] Figure 5 is a table showing the effect of viscosity modifiers on the stability of 175 mg / mL mAb1 incubated at 45°C for 14 days.aThe pH, SEUPLC and CEX-UPLC results for 'Raw Material' are the average values ​​for the Raw Material across all formulations. DETAILED DESCRIPTION

[0046] Before the present methods are described, it should be understood that the present invention is not limited to the particular experimental methods and conditions described, since those methods and conditions may vary. It should also be understood that the terminology used herein is intended to describe only particular embodiments, and is not intended to be limiting, since the scope of the present invention will be limited only by the appended claims.

[0047] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which the present invention pertains. As used herein, the term "about," when used in reference to a specific numerical value or range of values ​​quoted, means that the value may vary from the quoted value by no more than 1%. For example, as used herein, the expression "about 100" includes 99 and 101 and all intermediate values ​​(e.g., 99.1, 99.2, 99.3, 99.4, etc.). Although any methods and materials similar or equivalent to those described herein may be used in the practice or testing of the present invention, preferred methods and materials are now described. All publications mentioned herein are incorporated by reference in full.

[0048] As used herein, the expression pharmaceutical formulation means a combination of at least one ingredient. Petition 870220073975, dated 08 / 17 / 2022, p. 30 / 245 20 / 130 active ingredient (e.g., a small molecule, macromolecule, compound, etc., which is capable of exerting a biological effect on a human or non-human animal) and at least one inactive ingredient which, when combined with the active ingredient or one or more additional inactive ingredients, is suitable for therapeutic administration to a human or non-human animal. The term formulation, as used herein, means pharmaceutical formulation unless otherwise indicated. The present invention provides pharmaceutical formulations comprising at least one therapeutic polypeptide. According to certain embodiments of the present invention, the therapeutic polypeptide is an antibody or an antigen-binding fragment thereof that specifically binds to human programmed death protein-1 (PD-1).More specifically, the present invention includes pharmaceutical formulations comprising: (i) a human antibody that binds specifically to human PD-1; (ii) a histidine buffer; (iii) an organic cosolvent that is a nonionic surfactant; (iv) a stabilizer that is a carbohydrate; and, optionally, (v) a viscosity modifier that is an amino acid. The formulations included in the present invention are described in detail below. Antibodies that bind specifically to PD-1

[0049] The pharmaceutical formulations of the present invention may comprise a human antibody or an antigen-binding fragment thereof that binds specifically to human PD-1. As used herein, the term PD-1 means human programmed death protein-1. Anti-human PD-1 antibodies are described, for example, in US8008449, US8168757, US20110008369, US20130017199, US20130022595, US20150203579, and in documents WO2006121168, WO2009114335, WO2012145493, WO2013014668, WO2009101611, WO2015112800, EP2262837 and EP2504028. Petition 870220073975, dated 08 / 17 / 2022, page 31 / 245 21 / 130

[0050] The term antibody, as used herein, generally refers to immunoglobulin molecules comprising four polypeptide chains, two heavy (H) and two light (L) chains interconnected by disulfide bonds, as well as multimers thereof (e.g., IgM); however, immunoglobulin molecules consisting only of heavy chains (i.e., lacking light chains) are also included in the definition of the term antibody. Each heavy chain comprises a variable heavy chain region (abbreviated herein as HCVR or Vh) and a constant heavy chain region. The constant heavy chain region comprises three domains, CH1, CH2, and CH3. Each light chain comprises a variable light chain region (abbreviated herein as LCVR or Vl) and a constant light chain region. The constant light chain region comprises one domain (CL1).The Vh and Vl regions can be further subdivided into regions of hypervariability designated complementarity-determining regions (CDRs), interspersed with regions that are called more conserved structural regions (FRs). Each Vh and Vl region is composed of three CDRs and four FRs positioned, from the amino terminal to the carboxyl terminal, in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.

[0051] Unless specifically indicated otherwise, the term antibody, as used herein, should be understood to encompass complete antibody molecules as well as antigen-binding fragments thereof. The term antigen-binding portion or antigen-binding fragment of an antibody (or simply antibody portion or antibody fragment), as used herein, refers to one or more fragments of an antibody that retain the ability to bind specifically to human PD-1 or an epitope thereof.

[0052] An isolated antibody, as used herein, is intended to Petition 870220073975, dated 08 / 17 / 2022, p. 32 / 245 22 / 130 refers to an antibody that is substantially free of other antibodies that have different antigenic specificities (for example, an isolated antibody that binds specifically to human PD-1 is substantially free of antibodies that bind specifically to antigens other than human PD-1).

[0053] The term specifically binds or similar means that an antibody or antigen-binding fragment thereof 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 1x10-8M or greater. Methods for determining whether two molecules bind specifically are well known in the art and include, for example, equilibrium dialysis, surface plasmon resonance, and so forth. An isolated antibody that specifically binds to human PD-1 may, however, have cross-reactivity with other antigens, such as PD-1 molecules from other species (orthologs). In the context of the present invention, multispecific (e.g., bispecific) antibodies that bind to human PD-1 as well as to one or more additional antigens are considered to specifically bind to human PD-1.Furthermore, an isolated antibody may be substantially free of other cellular material or chemicals.

[0054] Examples of human anti-PD-1 antibodies that can be included in the pharmaceutical formulations of the present invention are presented in US20150203579 and WO2015112800, and whose descriptions are incorporated by reference in full.

[0055] According to certain embodiments of the present invention, the human anti-PD-1 antibody or antigen-binding fragment thereof comprises a heavy chain complementarity-determining region (HCDR) 1 of SEQ ID NO: 3, a Petition 870220073975, dated 08 / 17 / 2022, page 33 / 245 23 / 130 HCDR2 with SEQ ID NO: 4 and an HCDR3 with SEQ ID NO: 5. In certain embodiments, the human anti-PD-1 antibody or antigen-binding fragment thereof comprises an HCVR with SEQ ID NO: 1.

[0056] According to certain embodiments of the present invention, the human anti-PD-1 antibody or antigen-binding fragment thereof comprises a light chain complementarity-determining region (LCDR) 1 of SEQ ID NO: 6, an LCDR2 of SEQ ID NO: 7 and an LCDR3 of SEQ ID NO: 8. In certain embodiments, the human anti-PD-1 antibody or antigen-binding fragment thereof comprises an LCVR of SEQ ID NO: 2.

[0057] According to certain embodiments of the present invention, the human anti-PD-1 antibody or antigen-binding fragment thereof comprises an HCVR that has 90%, 95%, 98% or 99% sequence identity with SEQ ID NO: 1.

[0058] According to certain embodiments of the present invention, the human anti-PD-1 antibody or antigen-binding fragment thereof comprises an LCVR that has 90%, 95%, 98% or 99% sequence identity with SEQ ID NO: 2.

[0059] According to certain embodiments of the present invention, the human anti-PD-1 antibody or antigen-binding fragment thereof comprises an HCVR comprising an amino acid sequence of SEQ ID NO: 1 having no more than 5 amino acid substitutions.

[0060] According to certain embodiments of the present invention, the human anti-PD-1 antibody or antigen-binding fragment thereof comprises an LCVR comprising an amino acid sequence of SEQ ID NO: 2 having no more than 2 amino acid substitutions. Petition 870220073975, dated 08 / 17 / 2022, p. 34 / 245 24 / 130

[0061] Sequence identity can be measured using any method known in the art (e.g., GAP, BESTFIT, and BLAST).

[0062] The present invention also includes formulations comprising anti-PD-1 antibodies, wherein the anti-PD-1 antibodies comprise variants of any of the HCVR, LCVR and / or CDR amino acid sequences described herein that have one or more conservative amino acid substitutions. For example, the present invention includes formulations comprising anti-PD-1 antibodies having HCVR, LCVR and / or CDR amino acid sequences with, for example, 10 or fewer, 8 or fewer, 6 or fewer, 4 or fewer, etc. amino acid substitutions relative to any of the HCVR, LCVR and / or CDR amino acid sequences described herein.

[0063] In certain embodiments, the anti-PD1 antibody comprises an Fc region selected from the group consisting of human IgG1, IgG2, IgG3 and IgG4 isotypes.

[0064] The non-limiting exemplary antibody used in the Examples herein is referred to as mAb1. This antibody is also cited in US document 20150203579 as H2M7798N or H4H7798N and is also known as REGN2810 or cemiplimab. mAb1 (H4H7798N) comprises an HCVR / LCVR amino acid sequence pair having SEQ ID NOs: 1 / 2 and HCDR1-HCDR2-HCDR3 / LCDR1-LCDR2-LCDR3 domains represented by SEQ ID NOs: 3-55 / SEQ ID NOs: 6-7-8.

[0065] According to certain embodiments of the present invention, the human anti-PD-1 antibody or antigen-binding fragment thereof comprises a heavy chain of SEQ ID NO: 9 and a light chain of SEQ ID NO: 10.

[0066] It is well known in the art that terminal cleavage of Petition 870220073975, dated 08 / 17 / 2022, p. 35 / 245 25 / 130 amino acids can occur during antibody production (see, for example, Wang et al. 2007, J. Pharma Sci 96: 1-26). Therefore, in certain embodiments, the anti-PD-1 antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 11. SEQ ID NO: 11 comprises the heavy chain amino acid sequence in which the C-terminal lysine is absent from the amino acid sequence of SEQ ID NO: 9. In certain embodiments, the formulations of the present invention contain about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, about 98% or more of the anti-PD-1 antibody in which the C-terminal lysine is absent.

[0067] The amount of antibody or antigen-binding fragment thereof contained in the pharmaceutical formulations of the present invention may vary depending on the specific properties desired of the formulations, as well as the specific circumstances and purposes for which the formulations are intended to be used.In certain embodiments, pharmaceutical formulations are liquid formulations that may contain 5 ± 0.75 mg / mL to 250 ± 37.5 mg / mL of antibody; 10 ± 1.5 mg / mL to 240 ± 36 mg / mL of antibody; 20 ± 3.0 mg / mL to 230 ± 34.5 mg / mL of antibody; 25 ± 3.75 mg / mL to 240 ± 36 mg / mL of antibody; 50 ± 7.5 mg / mL to 230 ± 34.5 mg / mL of antibody; 60 ± 9 mg / mL to 240 ± 36 mg / mL of antibody; 70 ± 10.5 mg / mL to 230 ± 34.5 mg / mL antibody; 80 ± 12 mg / mL to 220 ± 33 mg / mL antibody; 90 ± 13.5 mg / mL to 210 ± 31.5 mg / mL antibody; 100 ± 15 mg / mL to 200 ± 30 mg / mL antibody; 110 ± 16.5 mg / mL to 190 ± 28.5 mg / mL antibody; 120 ± 18 mg / mL to 180 ± 27 mg / mL antibody; 130 ± 19.5 mg / mL to 170 ± 25.5 mg / mL antibody; 140 ± 21 mg / mL to 160 ± 24 mg / mL of antibody; 150 ± 22.5 mg / mL of antibody; or 175 ± 26.25 mg / mL. For example, the formulations of the present invention may comprise about 5 mg / mL; about 10. Petition 870220073975, dated 08 / 17 / 2022, page 36 / 245 26 / 130 mg / mL; approximately 15 mg / mL; approximately 20 mg / mL; approximately 25 mg / mL; approximately 30 mg / mL; approximately 35 mg / mL; approximately 40 mg / mL; approximately 45 mg / mL; approximately 50 mg / mL; approximately 55 mg / mL; approximately 60 mg / mL; approximately 65 mg / mL; approximately 70 mg / mL; approximately 75 mg / mL; approximately 80 mg / mL; approximately 85 mg / mL; approximately 90 mg / mL; approximately 95 mg / mL; approximately 100 mg / mL; approximately 105 mg / mL; approximately 110 mg / mL; approximately 115 mg / mL; approximately 120 mg / mL; approximately 125 mg / mL; approximately 130 mg / mL; approximately 135 mg / mL; approximately 140 mg / mL; approximately 145 mg / mL; approximately 150 mg / mL; approximately 155 mg / mL; approximately 160 mg / mL; approximately 165 mg / mL; approximately 170 mg / mL; approximately 175 mg / mL; approximately 180 mg / mL; approximately 185 mg / mL; approximately 190 mg / mL; approximately 195 mg / mL; approximately 200 mg / mL; approximately 205 mg / mL; approximately 210 mg / mL; approximately 215 mg / mL; approximately 220 mg / mL; approximately 225 mg / mL; approximately 230 mg / mL; approximately 235 mg / mL; approximately 240 mg / mL; approximately 245 mg / mL;or approximately 250 mg / mL of an antibody or an antigen-binding fragment thereof that binds specifically to human PD-1. EXCIPIENTS AND pH

[0068] The pharmaceutical formulations of the present invention comprise one or more excipients. The term excipient, as used herein, means any non-therapeutic agent added to the formulation to impart a desired consistency, viscosity or stabilizing effect.

[0069] In certain embodiments, the pharmaceutical formulation of the invention comprises at least one organic cosolvent in a type and amount that stabilizes the human anti-PD-1 antibody under conditions of agitation or rough handling such as, for example, vortexing. In some embodiments, what is meant by stabilizing is the prevention of the formation of more than 3% of aggregated antibody from the total amount of antibody (on a molar basis) during the course of rough handling. In some embodiments, the Petition 870220073975, dated 08 / 17 / 2022, page 37 / 245 27 / 130 Rough manipulation is to vortex a solution containing the antibody and the organic cosolvent for about 60 minutes or about 120 minutes.

[0070] In certain embodiments, the organic cosolvent is a nonionic surfactant, such as an alkyl (poly)ethylene oxide. Specific nonionic surfactants that may be included in the formulations of the present invention include, for example, polysorbates, such as polysorbate 20, polysorbate 28, polysorbate 40, polysorbate 60, polysorbate 65, polysorbate 80, polysorbate 81, polysorbate 81 and polysorbate 85; poloxamers, such as poloxamer 181, poloxamer 188, poloxamer 407; or polyethylene glycol (PEG). Polysorbate 20 is also known as TWEEN 20, sorbitan monolaurate and polyoxyethylene sorbitan monolaurate. Poloxamer 188 is also known as Pluronic F68.

[0071] The amount of nonionic surfactant contained in the pharmaceutical formulations of the present invention may vary depending on the specific properties desired of the formulations, as well as the specific circumstances and purposes for which the formulations are to be used. In certain embodiments, the formulations may contain 0.01% ± 0.005% to 0.5% ± 0.25% of surfactant. For example, the formulations of the present invention may comprise 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%; around 0.13%; around 0.14%; about 0.15%; around 0.16%; around 0.17%; around 0.18%; about 0.19%; around 0.20%; around 0.21%; around 0.22%; about 0.23%; around 0.24%; around 0.25%; around 0.26%; about 0.27%; around 0.28%; around 0.29%; around 0.30%; about 0.35%; around 0.40%; around 0.45%; around 0.46%; about Petition 870220073975, dated 08 / 17 / 2022, page 38 / 245 28 / 130 0.47%; approximately 0.48%; approximately 0.49%; approximately 0.50%; approximately 0.55%; or approximately 0.575% of polysorbate 20 or polysorbate 80.

[0072] The pharmaceutical formulations of the present invention may also comprise one or more stabilizers in a type and amount that stabilizes the human anti-PD-1 antibody under thermal stress conditions. In some embodiments, what is meant by stabilization is the maintenance of more than about 91% of the antibody in a native conformation when the solution containing the antibody and the thermal stabilizer is maintained at about 45°C for up to about 28 days. In some embodiments, what is meant by stabilization is the maintenance of less than about 6% of the aggregated antibody when the solution containing the antibody and the thermal stabilizer is maintained at about 45°C for up to about 28 days. As used herein, native means the primary form of the antibody by size exclusion, which is generally an intact monomer of the antibody. The term native also refers to the non-aggregated and non-degraded form of the antibody.

[0073] In certain embodiments, the thermal stabilizer is a sugar, such as sucrose, the amount of which in the formulation may vary depending on the specific circumstances and the intended purposes for which the formulation is used. In certain embodiments, the formulations may contain about 1% to about 15% sugar; about 2% to about 14% sugar; about 3% to about 13% sugar; about 4% to about 12% sugar; about 5% to about 12% sugar; about 6% to about 11% sugar; about 7% to about 10% sugar; about 8% to 11% sugar; or about 9% to 11% sugar. For example, the pharmaceutical formulations of the present invention may comprise 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 approximately Petition 870220073975, dated 08 / 17 / 2022, p. 39 / 245 29 / 130 of 14% ± 2.8% sugar (e.g., sucrose).

[0074] The pharmaceutical formulations of the present invention may also comprise a buffer or buffer system that serves to maintain a stable pH and helps stabilize the human anti-PD-1 antibody. The term buffer, as used herein, denotes a pharmaceutically acceptable buffer that maintains a stable pH or resists changes in the pH of the solution. In preferred embodiments, the buffer comprises histidine. In the context of the present invention, histidine buffer or buffer comprising histidine is a buffer comprising the amino acid histidine. Examples of histidine buffers include histidine chloride, histidine acetate, histidine phosphate, and histidine sulfate. In a preferred embodiment, the histidine buffer is prepared by dissolving L-histidine and L-histidine hydrochloride (e.g., as the monohydrate) in a defined amount and ratio. In another 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 invention. In some embodiments, stabilization is meant where less than 4.5% ± 0.5% of the antibody is aggregated when the solution containing the antibody and buffer is maintained at about 45°C for up to about 28 days. In some embodiments, stabilization is meant where less than 3% ± 0.5% or less than 2.5% ± 0.5% of the antibody is aggregated when the solution containing the antibody and buffer is maintained at about 37°C for up to 28 days. In some embodiments, what is meant by stabilization is where at least 93% ± 0.5% or at least 94% ± 0.5% of the antibody is in its native conformation, as determined by size exclusion chromatography when the solution containing the antibody and buffer is maintained at about 45°C for up to about 28 days. In some embodiments, what... Petition 870220073975, dated 08 / 17 / 2022, p. 40 / 245 30 / 130 is defined as stabilization where at least 94% ± 0.5% or at least 95% ± 0.5% of the antibody is in its native conformation, as determined by size exclusion chromatography when the solution containing the antibody and buffer is maintained at approximately 37°C for up to approximately 28 days. Native conformation refers to the fraction of antibody that is not aggregated or degraded. This is usually determined by an assay that measures the relative size of the antibody entity, such as a size exclusion chromatography assay. Unaggregated and undegraded antibody elutes in a fraction equivalent to the native antibody and is usually the main elution fraction. Aggregated antibody elutes in a fraction indicating a larger size than the native antibody. Degraded antibody elutes in a fraction indicating a smaller size than the native antibody.

[0075] In some embodiments, stabilization is understood to mean that at least 35% ± 0.5% of the antibody is in its primary charge form, as determined by cation-exchange chromatography when the solution containing the antibody and buffer is maintained at about 45°C for up to about 28 days. In some embodiments, stabilization is understood to mean that at least 46% ± 0.5% or at least 39% ± 0.5% of the antibody is in its primary charge form, as determined by cation-exchange chromatography when the solution containing the antibody and buffer is maintained at about 37°C for up to about 28 days. The term "main charge" or "main charge form" refers to the fraction of antibody that elutes from an ion-exchange resin at the main peak, which is usually flanked by more basic peaks on one side and more acidic peaks on the other.

[0076] The pharmaceutical formulations of the present invention may have a pH of about 5.2 to about 6.4. For example, the formulations of Petition 870220073975, dated 08 / 17 / 2022, p. 41 / 245 31 / 130 of the present invention may have a pH of approximately 5.5; approximately 5.6; approximately 5.7; approximately 5.8; approximately 5.9; approximately 6.0; approximately 6.1; approximately 6.2; approximately 6.3; approximately 6.4; or approximately 6.5. In some embodiments, the pH is 6.0 ± 0.4; 6.0 ± 0.3; 6.0 ± 0.2; 6.0 ± 0.1; approximately 6.0; or 6.0.

[0077] In some embodiments, the buffer or buffer system comprises at least one buffer that has a buffering range that totally or partially overlaps the pH range 5.5 - 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 histidine at 10 mM ± 2 mM at a pH of 6.0 ± 0.3. In preferred 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 another 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.

[0078] The pharmaceutical formulations of the present invention may also comprise one or more excipients that serve to maintain a reduced viscosity or decrease the viscosity of formulations containing a high concentration of anti-PD-1 antibody pharmaceutical substance (for example, generally > 150 mg / mL of antibody). In certain embodiments, the viscosity modifier is a Petition 870220073975, dated 08 / 17 / 2022, page 42 / 245 32 / 130 amino acid. In one embodiment, the amino acid is proline. In one embodiment, the pharmaceutical formulation of the present invention contains proline, preferably as L-proline, at a concentration of 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, or 5%. The term proline is used interchangeably with L-proline throughout the present invention. In some embodiments, the formulation comprises proline in an amount sufficient to maintain the viscosity of the liquid formulation at less than 20 ± 3 cPoise, less than 15 ± 2.25 cPoise, or less than 11 ± 1.65 cPoise. In some embodiments, the formulation comprises proline in an amount sufficient to maintain the viscosity equal to or less than 15 ± 2.25 cPoise. In certain embodiments, the formulations may contain proline at about 1% to about 5%; proline at about 2% to about 4%; or proline at about 3%.For example, the pharmaceutical formulations of the present invention may comprise proline at 1% ± 0.2%; 1.5% ± 0.3%; 2% ± 0.4%; 2.5% ± 0.5%; 3% ± 0.6%; 3.5% ± 0.7%; 4% ± 0.8%; 4.5% ± 0.9%; or about 5% ± 1%.

[0079] During the antibody purification process, it may be desirable or necessary to exchange one buffer for another to obtain appropriate excipient concentrations, antibody concentrations, pH, etc. Buffer exchange can be performed, for example, by ultrafiltration / diafiltration (UF / DF) using, for example, a semipermeable tangential flow filtration membrane. The use of such techniques, however, has the potential to cause the Gibbs-Donnan effect [Bolton et al., 2011, Biotechnol. Prog. 27 (1): 140-152]. The accumulation of positive charge on the product side of the membrane during protein concentration is electrically counterbalanced by the preferential movement of positive ions to the opposite side of the membrane. The potential consequence of this phenomenon is that the final concentrations of certain components (e.g., Petition 870220073975, dated 08 / 17 / 2022, page 43 / 245 33 / 130 histidine, L-proline, etc.) may be lower than the intended target concentrations of these components due to electrostatic repulsion of the positively charged diafiltration buffer excipients towards the positively charged antibody protein during the UF / DF step. Thus, the present invention includes formulations in which the concentration of, for example, histidine and / or L-proline, varies between the amounts or ranges cited herein due to the Gibbs-Donnan effect.

[0080] Volume exclusion describes the behavior of highly concentrated samples in which a significant portion of the total solution volume is absorbed by the solute, especially large molecules such as proteins, excluding the solvent from this space. This then decreases the total volume of solvent available for other solutes to be dissolved, which can result in uneven partitioning across the ultrafiltration membrane. Thus, the present invention includes formulations in which the concentration of, for example, histidine and / or L-proline, can vary relative to the amounts or ranges cited herein by virtue of the volume exclusion effect.

[0081] During the manufacture of the formulations of the present invention, variations in the formulation composition may occur. These variations may include the concentration of the active ingredient, the concentration of excipients, and / or the pH of the formulation. Since changes in any of these parameters can potentially affect the stability or potency of the drug, proven acceptable range (PAR) studies were conducted to evaluate whether variations in composition, within defined limits, would affect the stability or potency of the antibody. Therefore, the present invention includes formulations comprising anti-PD-1 antibodies that are stable and retain potency with variations of up to 50% in excipient concentration. For example, antibody formulations are included herein. Petition 870220073975, dated 08 / 17 / 2022, page 44 / 245 34 / 130 anti-PD-1 wherein the stability and potency of said formulations are not affected by a variation of ± 10%, ± 20%, ± 30%, ± 30%, ± 40% or ± 50% in the concentration of antibody, sucrose, histidine buffer and / or polysorbate. Stability and viscosity of pharmaceutical formulations.

[0082] The pharmaceutical formulations of the present invention typically exhibit high levels of stability. The term stable, as used herein in reference to pharmaceutical formulations, means that the antibodies in the pharmaceutical formulations retain an acceptable degree of chemical structure or biological function after storage under defined conditions. A formulation may be stable even if the antibody contained therein does not retain 100% of its chemical structure or biological function after storage for a defined period of time. Under certain circumstances, the maintenance of approximately 90%, 95%, 96%, 97%, 98%, or 99% of the structure or function of an antibody after storage for a defined period may be considered stable.

[0083] Stability can be measured, inter alia, by determining the percentage of native antibody that remains in the formulation after storage for a defined period of time at a defined temperature. The percentage of native antibody can be determined, inter alia, by size exclusion chromatography (e.g., ultra-performance liquid chromatography with size exclusion [SE-UPLC]), wherein native means non-aggregated and non-degraded. An acceptable degree of stability, as such phrase is used herein, means that at least 90% of the native form of the antibody can be detected in the formulation after storage for a defined period of time at a given temperature. In certain embodiments, at least about 90%, 91%, 92%, 93% Petition 870220073975, dated 08 / 17 / 2022, page 45 / 245 35 / 130%, 94%, 95%, 96%, 97%, 98%, 99% or 100% of the native antibody form can be detected in the formulation after storage for a defined period at a defined temperature. The defined period after which stability is measured can be at least 14 days, at least 28 days, at least 1 month, at least 2 months, at least 3 months, at least 4 months, 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 defined temperature at which the pharmaceutical formulation can be stored when assessing stability can be any temperature from approximately -80°C to 45°C, for example, storage at approximately -80°C, approximately -30°C, approximately -20°C, approximately 0°C, approximately 4°-8°C, approximately 5°C, approximately 25°C, approximately 35°C, approximately 37°C, or approximately 45°C.For example, a pharmaceutical formulation can be considered stable if, after 6 months of storage at 5°C, more than approximately 95%, 96%, 97%, or 98% of the native antibody is detected by SE-UPLC. A pharmaceutical formulation can also be considered stable if, after 6 months of storage at 25°C, more than approximately 95%, 96%, 97%, or 98% of the native antibody is detected by SE-UPLC. A pharmaceutical formulation can also be considered stable if, after 28 days of storage at 45°C, more than approximately 89%, 90%, 91%, 92%, 93%, 94%, 95%, or 96% of the native antibody is detected by SE-UPLC. A pharmaceutical formulation can also be considered stable if, after 12 months of storage at -20°C, more than approximately 96%, 97%, or 98% of the native antibody is detected by SE-UPLC. A pharmaceutical formulation can also be considered stable if, after 12 months of storage at -30°C, more than approximately 96%, 97%, or 98% of the native antibody is detected by SE-UPLC. Petition 870220073975, dated 08 / 17 / 2022, page 46 / 245 36 / 130% or 98% of the native antibody is detected by SE-UPLC. A pharmaceutical formulation can also be considered stable if, after 12 months of storage at -80°C, more than 96%, 97%, or 98% of the native antibody is detected by SE-UPLC.

[0084] Stability can be measured, inter alia, by determining the percentage of antibody that forms in an aggregate within the formulation after storage for a defined period of time at a defined temperature, wherein stability is inversely proportional to the percentage of aggregate formed. The percentage of aggregated antibody can be determined, inter alia, by size exclusion chromatography (e.g., ultra-performance liquid chromatography with size exclusion [SE-UPLC]). An acceptable degree of stability, as this sentence is used herein, means that at most 5% of the antibody is in an aggregated form (also designated as high molecular weight form HMW) detected in the formulation after storage for the defined amount of time at a given temperature.In certain embodiments, an acceptable degree of stability means that at most about 5%, 4%, 3%, 2%, 1%, 0.5% or 0.1% of the antibody can be detected in an aggregate in the formulation after storage for a defined period of time at a defined temperature. The defined period of time after which stability is measured may 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 may be stored when stability is assessed may be any temperature from about -80°C. Petition 870220073975, dated 08 / 17 / 2022, page 47 / 245 37 / 130 up to approximately 45°C, for example, storage at approximately -80°C, approximately -30°C, approximately -20°C, approximately 0°C, approximately 4-8°C, approximately 5°C, approximately 25°C, approximately 35°C, approximately 37°C or approximately 45°C. For example, a pharmaceutical formulation may be considered stable if, after 12 months of storage at 5°C, less than 2%, 1%, 0.5% or 0.1% of the antibody is detected in aggregated form. A pharmaceutical formulation may also be considered stable if, after three months of storage at 25°C, less than approximately 4%, 3%, 2%, 1%, 0.5% or 0.1% of the antibody is detected in aggregated form. A pharmaceutical formulation can also be considered stable if, after 28 days of storage at 45°C, less than approximately 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or 0.5% of the antibody is detected in aggregated form.A pharmaceutical formulation may also be considered stable if, after three months of storage at -20°C, -30°C or -80°C, less than about 3%, 2%, 1%, 0.5% or 0.1% of the antibody is detected in an aggregated form.

[0085] Stability can be measured, inter alia, by determining the percentage of antibody that migrates in a more acidic fraction during ion exchange (acid form) than in the main antibody fraction (main charge form), wherein stability is inversely proportional to the fraction of antibody in the acid form. Although not wishing to be limited by theory, antibody deamidation may cause the antibody to become more negatively charged and therefore more acidic relative to the non-deamidated antibody (see, for example, Robinson, N., Protein Deamidation, PNAS, April 16, 2002, 99(8): 5283-5288).The percentage of acidified antibody can be determined, inter alia, by means of ion exchange chromatography (e.g., cation exchange ultra-high performance liquid chromatography [CEX-UPLC]). An acceptable degree of stability, as that phrase is used here, means that at most 45% of the antibody is acidified. Petition 870220073975, dated 08 / 17 / 2022, page 48 / 245 38 / 130 in a more acidic form detected in the formulation after storage for a defined period of time at a defined temperature. In certain embodiments, an acceptable degree of stability means that at most about 40%, 35%, 30%, 25%, 20%, 15%, 10%, 5%, 4%, 3%, 2%, 1%, 0.5% or 0.1% of the antibody can be detected in an acidic form in the formulation after storage for a defined period of time at a defined temperature. In one embodiment, an acceptable degree of stability means that less than 30%, 25%, 20%, 15%, 10%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the antibody can be detected in an acidic form in the formulation after storage for a defined period of time.The defined period 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 assessing 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 4-8°C, about 5°C, about 25°C or about 45°C.For example, a pharmaceutical formulation can be considered stable if, after three months of storage at -80°C, 30°C or -20°C, less than approximately 30%, 29%, 28%, 27%, 26%, 25%, 24%, 23%, 22%, 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5% or 0.1% of the antibody is in a more acidic form. A pharmaceutical formulation can also be considered stable if, after six months of storage at 5°C, less than 32%, 31%, 30% Petition 870220073975, dated 08 / 17 / 2022, page 49 / 245 39 / 130%, 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. A pharmaceutical formulation can also be considered stable if, after six months of storage at 25°C, less than 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.A pharmaceutical formulation can also be considered stable if, after 28 days of storage at 45°C, less than approximately 49%, 48%, 47%, 46%, 45%, 44%, 43%, 43%, 42%, 41%, 40%, 39%, 38%, 37%, 36%, %, 34%, 33%, 32%, 31%, 30%, 29%, 28%, 27%, 26%, 25%, %, 23%, 22%, 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, %, 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.

[0086] Other methods may be used to evaluate the stability of the formulations of the present invention, such as, for example, differential scanning calorimetry (DSC) to determine thermal stability, controlled stirring to determine mechanical stability, and absorbance at about 350 nm or about 405 nm to determine the turbidity of the solution. For example, a formulation of the present invention may be considered stable if, after 6 or more months of storage at about 5°C to about 25°C, the change in OD405 of the formulation is less than about 0.05 (e.g., 0.04, 0.03, 0.02, 0.01 or less) relative to the OD405 of the formulation at time zero.

[0087] Measuring the biological activity or binding affinity of the antibody to its target can also be used to assess the Petition 870220073975, dated 08 / 17 / 2022, page 50 / 245 40 / 130 Stability. For example, a formulation of the present invention may be considered stable if, after storage, for example, at 5°C, 25°C, 45°C, etc., for a defined period of time (for example, 1 to 12 months), the anti-PD-1 antibody contained in the formulation binds to PD-1 with an affinity that is at least 90%, 95% or more of the antibody binding affinity before said storage. The binding affinity may be determined, for example, by ELISA or surface plasmon resonance. Biological activity may be determined by a PD-1 activity assay such as, for example, contact of a cell expressing PD-1 with the formulation comprising the anti-PD-1 antibody. Antibody binding to a cell may be measured directly, for example, via FACS analysis. Alternatively, the downstream activity of the PD-1 system may be measured in the presence of the antibody and compared with the activity of the PD-1 system in the absence of antibody.In some forms, PD-1 may be endogenous to the cell. In other forms, PD-1 may be ectopically expressed in the cell.

[0088] Additional methods for evaluating the stability of an antibody in the formulation are demonstrated in the Examples presented below.

[0089] The liquid pharmaceutical formulations of the present invention may, in certain embodiments, exhibit low to moderate viscosity levels. Viscosity, as used herein, may be kinematic viscosity or absolute viscosity. Kinematic viscosity is a measure of the resistive flow of a fluid under the influence of gravity. When two fluids of equal volume are placed in identical capillary viscometers and allowed to flow by gravity, a viscous fluid takes longer than a less viscous fluid to flow through the capillary. For example, if one fluid takes 200 seconds to complete its flow and another fluid takes 400 seconds, the latter Petition 870220073975, dated 08 / 17 / 2022, p. 51 / 245 41 / 130 fluid is twice as viscous as the first on a kinematic viscosity scale. Absolute viscosity, sometimes called dynamic or simple viscosity, is the product of kinematic viscosity and fluid density (Absolute viscosity = kinematic viscosity x density). The dimension of kinematic viscosity is L² / T, where L is length and T is time. In general, kinematic viscosity is expressed in centistokes (cSt). The SI unit of kinematic viscosity is mm² / s, which is 1 cSt. Absolute viscosity is expressed in centipoise (cP) units. The SI unit of absolute viscosity is millipascal-second (mPa-s), where 1 cP = 1 mPa-s.

[0090] As used herein, a low viscosity level, with reference to a fluid formulation of the present invention, will exhibit an absolute viscosity of less than about 20 cPoise (cP). For example, a fluid formulation of the invention will be considered to have a low viscosity if, when measured using standard viscosity measurement techniques, 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 used herein, a moderate viscosity level, with reference to a fluid formulation of the present invention, will exhibit an absolute viscosity between about 35 cP and about 20 cP.For example, a fluid formulation of the invention will be considered to have a moderate viscosity if, when measured using standard viscosity measurement techniques, 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 15.1 cP. Petition 870220073975, dated 08 / 17 / 2022, page 52 / 245 42 / 130

[0091] As illustrated in the examples below, the present inventors have made the surprising finding that low viscosity liquid formulations comprising high concentrations of a human anti-PD-1 antibody (e.g., from about 50 mg / mL to 250 mg / mL) can be obtained by formulating the antibody with proline at about 1% to about 5% and sucrose at about 5%. Such formulations are stable to stress during handling and storage at temperatures ranging from 45°C to -80°C (shown here) and have low viscosity (having viscosity ranging from 7 to 15 cP). EXEMPLARY FORMULATIONS

[0092] According to one aspect of the present invention, the pharmaceutical formulation is a stable, low-viscosity, generally physiologically isotonic liquid formulation comprising: (i) a human antibody that specifically binds to human PD-1 (e.g., H4H7798N) at a concentration of up to 250 mg / mL ± 45 mg / mL; (ii) a histidine buffer system that provides sufficient buffering at a pH of about 6.0 ± 0.3; (iii) an organic cosolvent that protects the structural integrity of the antibody; (iv) a thermal stabilizer that is a sugar; and (v) a viscosity modifier that is an amino acid that serves to maintain manageable viscosity for injection in a volume convenient for subcutaneous administration.

[0093] According to one embodiment, the stable low-viscosity pharmaceutical formulation comprises: (i) a human IgG4 antibody that specifically binds to human PD-1 and comprises an HCDR1 with SEQ ID NO: 3, an HCDR2 with SEQ ID NO: 4, an HCDR3 with SEQ ID NO: 5, an LCDR1 with SEQ ID NO: 6, an LCDR2 with SEQ ID NO: 7 and an LCDR3 with SEQ ID NO: 8 at a concentration of up to 200 mg / mL ± 30 mg / mL; (ii) 10 mM ± 2 mM histidine buffer buffering at pH 6.0 ± 0.3; (iii) 0.2% w / v ± 0.1% polysorbate 80 Petition 870220073975, dated 08 / 17 / 2022, p. 53 / 245 43 / 130 w / v; (iv) sucrose at 5% ± 1% w / v; and (v) L-proline at 1.5% (w / v) ± 0.3%.

[0094] According to one embodiment, the stable low-viscosity pharmaceutical formulation comprises: (i) a human IgG4 antibody that specifically binds to human PD-1 and comprises an HCDR1 of SEQ ID NO: 3, an HCDR2 of SEQ ID NO: 4, an HCDR3 of SEQ ID NO: 5, an LCDR1 of SEQ ID NO: 6, an LCDR2 of SEQ ID NO: 7 and an LCDR3 of SEQ ID NO: 8 at a concentration of 175 mg / mL ± 26.25 mg / mL; (ii) 10 mM ± 2 mM histidine buffer that buffers at a pH of 6.0 ± 0.3; (iii) polysorbate 80 at 0.2% w / v ± 0.1% w / v; (iv) sucrose at 5% ± 1% w / v; and (v) L-proline at 1.5% (w / v) ± 0.3%.

[0095] According to one embodiment, the stable low-viscosity pharmaceutical formulation comprises: (i) a human IgG4 antibody that specifically binds to human PD-1 and comprising an HCDR1 of SEQ ID NO: 3, an HCDR2 of SEQ ID NO: 4, an HCDR3 of SEQ ID NO: 5, an LCDR1 of SEQ ID NO: 6, an LCDR2 of SEQ ID NO: 7 and an LCDR3 of SEQ ID NO: 8 at a concentration of 150 mg / mL ± 22.5 mg / mL; (ii) 10 mM ± 2 mM histidine buffer buffering at pH 6.0 ± 0.3; (iii) 0.2% w / v ± 0.1% w / v polysorbate 80; (iv) 5% ± 1% w / v sucrose; and (v) L-proline at 1.5% (w / v) ± 0.3%.

[0096] According to one embodiment, the stable low-viscosity pharmaceutical formulation comprises: (i) a human IgG4 antibody that specifically binds to human PD-1 and comprising an HCDR1 of SEQ ID NO: 3, an HCDR2 of SEQ ID NO: 4, an HCDR3 of SEQ ID NO: 5, an LCDR1 of SEQ ID NO: 6, an LCDR2 of SEQ ID NO: 7 and an LCDR3 of SEQ ID NO: 8 at a concentration of 100 mg / mL ± 15 mg / mL; (ii) 10 mM ± 2 mM histidine buffer buffering at pH 6.0 ± 0.3; (iii) 5% w / v ± 1% w / v sucrose; (iv) 0.2% w / v ± 0.1% polysorbate 80; and L-proline at 1.5% (w / v) ± 0.3%.

[0097] According to one embodiment, the stable low-viscosity pharmaceutical formulation comprises: (i) an IgG4 Petition 870220073975, dated 08 / 17 / 2022, p. 54 / 245 44 / 130 human that specifically binds to human PD-1 and comprising an HCDR1 of SEQ ID NO: 3, an HCDR2 of SEQ ID NO: 4, an HCDR3 of SEQ ID NO: 5, an LCDR1 of SEQ ID NO: 6, an LCDR2 of SEQ ID NO: 7 and an LCDR3 of SEQ ID NO: 8 at a concentration of 50 mg / mL ± 7.5 mg / mL; (ii) 10 mM ± 2 mM histidine buffer buffering at pH 6.0 ± 0.3; (iii) 5% w / v ± 1% w / v sucrose; (iv) 0.2% w / v ± 0.1% polysorbate 80; and 1.5% (w / v) ± 0.3% L-proline.

[0098] According to one embodiment, the stable low-viscosity pharmaceutical formulation comprises: (i) a human IgG4 antibody that specifically binds to human PD-1 and comprising an HCDR1 of SEQ ID NO: 3, an HCDR2 of SEQ ID NO: 4, an HCDR3 of SEQ ID NO: 5, an LCDR1 of SEQ ID NO: 6, an LCDR2 of SEQ ID NO: 7 and an LCDR3 of SEQ ID NO: 8 at a concentration of 25 mg / mL ± 3.75 mg / mL; (ii) 10 mM ± 2 mM histidine buffer buffering at a pH of 6.0 ± 0.3; (iii) 5% w / v ± 1% w / v sucrose; (iv) 0.2% w / v ± 0.1% polysorbate 80; and L-proline at 1.5% (w / v) ± 0.3%.

[0099] Additional non-limiting examples of pharmaceutical formulations covered by the present invention are presented elsewhere herein, including the Working Examples presented below. CONTAINERS AND METHODS OF ADMINISTRATION

[0100] The pharmaceutical formulations of the present invention may be contained within any suitable container for storing drugs and other therapeutic compositions. For example, the pharmaceutical formulations may be contained in a sealed and sterilized plastic or glass container with a defined volume, such as a vial, ampoule, syringe, cartridge, or bottle. Different types of containers may be used to contain the formulations of the present invention, including, for example, transparent and opaque (e.g., amber) glass or plastic containers. Petition 870220073975, dated 08 / 17 / 2022, page 55 / 245 45 / 130 Similarly, any type of syringe can be used to contain or administer the pharmaceutical formulations of the present invention.

[0101] The pharmaceutical formulations of the present invention may be contained within ordinary tungsten syringes or low-tungsten syringes. As will be appreciated by those skilled in the art, the manufacturing process for glass syringes generally involves the use of a hot tungsten rod that functions to pierce the glass, thereby creating an orifice from which liquids can be extracted 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 may be used to reduce the amount of tungsten in the syringe. As used herein, the term ordinary tungsten means that the syringe contains more than or equal to 500 parts per billion (ppb) of tungsten. The term low-tungsten means that the syringe contains less than 500 ppb of tungsten.For example, a low-tungsten syringe, according to the present invention, 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 or less ppb of tungsten.

[0102] The rubber plungers used in syringes and the rubber stoppers used to close the openings of vials can be coated to prevent contamination of the medicinal contents of the syringe or vial or to preserve its stability. Thus, the pharmaceutical formulations of the present invention, according to certain embodiments, can be contained within a syringe comprising a coated plunger or within a vial that is sealed with a coated rubber stopper. For example, the plunger or stopper can be coated with a fluorocarbon film. Examples of coated stoppers or plungers suitable for use with vials and syringes that Petition 870220073975, dated 08 / 17 / 2022, p. 56 / 245 Examples of pharmaceutical formulations of the present invention, such as those contained in US 4,997,423; US 5,908,686; US 6,286,699; US 6,645,635; and US 7,226,554, are mentioned, the contents of which are incorporated herein by reference in their entirety. Specific exemplary rubber-coated stoppers and plungers that may be used in the context of the present invention are commercially available under the trade name FluroTec®, available from West Pharmaceutical Services, Inc. (Lionville, PA). FluroTec® is an example of a fluorocarbon coating used to minimize or prevent drug adhesion to rubber surfaces.

[0103] According to certain embodiments of the present invention, pharmaceutical formulations may be contained within a low-tungsten syringe comprising a fluorocarbon-coated plunger.

[0104] Pharmaceutical formulations can be administered to a patient via parenteral routes, such as injections (e.g., subcutaneous, intravenous, intramuscular, intraperitoneal, etc.) or percutaneous, mucosal, nasal, pulmonary, or oral routes. Numerous reusable pen or autoinjector delivery devices can be used to subcutaneously administer the pharmaceutical formulations of the present invention. Examples include, among others, AUTOPEN™ (Owen Mumford, Inc., Woodstock, United Kingdom), the DISETRONIC™ pen (Disetronic Medical Systems, Bergdorf, Switzerland), the HUMALOG MIX 75 / 25™ pen, the HUMALOG™ pen, the HUMALIN 70 / PEN 30™ pen (Eli Lilly and Co., Indianapolis, IN), NOVOPEN™ I, II and III (Novo Nordisk, Copenhagen, Denmark), NOVOPEN JUNIOR™ (Novo Nordisk, Copenhagen, Denmark), the BD™ pen (Becton Dickinson, Franklin Lakes, NJ), OPTIPEN™, OPTIPEN PRO™, OPTIPEN STARLET™ and OPTICLIK™ (Sanofi Aventis, Frankfurt, Germany). Examples of devices of Petition 870220073975, dated 08 / 17 / 2022, page 57 / 245 47 / 130 Disposable pen-type or autoinjector administration devices with applications for the subcutaneous administration of a pharmaceutical composition of the present invention include, among others, the SOLOSTAR™ pen (Sanofi-Aventis), FLEXPEN™ (Novo Nordisk) and KWIKPEN™ (Eli Lilly), the SURECLICK™ autoinjectors (Amgen, Thousand Oaks, CA), PENLET™ (Haselmeier, Stuttgart, Germany), EPIPENT™ (Dey, LP) and the HUMIRA™ pen (Abbott Labs, Abbott Park, IL).

[0105] The use of a microinfuser to administer the pharmaceutical formulations of the present invention is also considered herein. As used herein, the term microinfuser means a subcutaneous delivery device designed to slowly administer large volumes (e.g., up to about 2.5 mL or more) of a therapeutic formulation over an extended period of time (e.g., about 10, 15, 20, 25, 30 or more minutes). See, for example, US 6,629,949; US 6,659,982; and Meehan et al., J. Controlled Release 46: 107-116 (1996). Microinfusers are particularly useful for administering large doses of therapeutic proteins contained in high concentrations (e.g., about 100, 125, 150, 175, 200 or more mg / mL) or viscous solutions.

[0106] In certain embodiments, the stable liquid pharmaceutical formulation of any of the foregoing aspects is contained in a sterile glass vial and is administered as an IV infusion.

[0107] In one embodiment, the container is a 20 mL clear borosilicate glass bottle of type 1. In certain embodiments, the container is a 2 mL, 5 mL or 10 mL borosilicate glass bottle of type 1 with a chlorobutyl stopper, with a FluroTec® coating.

[0108] In one embodiment, the liquid pharmaceutical formulation of the present invention comprising about 25 mg / mL or 50 mg / mL Petition 870220073975, dated 08 / 17 / 2022, p. 58 / 245 48 / 130 mAbl is administered intravenously and may be contained in a glass vial.

[0109] In certain embodiments, the present invention provides an autoinjector comprising any of the liquid formulations described herein. In some embodiments, the present invention provides an autoinjector comprising a stable liquid formulation comprising about 50 mg / mL, about 100 mg / mL, about 150 mg / mL or about 175 mg / mL of mAb1, histidine at about 10 mM at a pH of about 6.0, sucrose at about 5%, proline at about 1.5% and polysorbate 80 at about 0.2%.

[0110] In certain embodiments, the present invention provides a pre-filled syringe comprising any of the liquid formulations described herein. In some embodiments, the present invention provides a pre-filled syringe comprising a stable liquid formulation comprising about 50 mg / mL, about 100 mg / mL, about 150 mg / mL or about 175 mg / mL of mAb1, histidine at about 10 mM at a pH of about 6.0, sucrose at about 5%, proline at about 1.5% and polysorbate 80 at about 0.2%. In certain embodiments, the syringe is a 1 mL or 2.25 mL glass syringe pre-filled with a 27-gauge thin-walled needle, a fluorocarbon-coated rubber plunger and a rubber needle guard.

[0111] In one embodiment, the liquid pharmaceutical formulation containing approximately 175 mg / mL ± 26.25 mg / mL of mAb1 is administered in a volume of approximately up to 2 mL in a pre-filled syringe. In certain embodiments, the syringe is a 1 mL or 2.25 mL glass syringe pre-filled with a 27-gauge thin-walled needle, a fluorocarbon-coated rubber plunger, and a rubber needle guard. In another embodiment, the syringe is a 1 mL WIPO long glass syringe fitted with a 27-gauge needle, a Petition 870220073975, dated 08 / 17 / 2022, page 59 / 245 49 / 130 FM27 rubber needle guard and a 4023 / 50 FluroTec® coated rubber plunger.

[0112] In one embodiment, the liquid pharmaceutical formulation containing approximately 150 mg / mL ± 22.5 mg / mL anti-DP-1 antibody is administered in a volume of up to approximately 2 mL in a pre-filled syringe. In one embodiment, the syringe is a 1 mL or 2.25 mL glass syringe pre-filled with a 27-gauge thin-walled needle, a fluorocarbon-coated rubber plunger, and a rubber needle guard. In another embodiment, the syringe is a 1 mL WIPO long glass syringe fitted with a 27-gauge needle, an FM27 rubber needle guard, and a FluroTec®-coated 4023 / 50 rubber plunger. THERAPEUTIC USES OF PHARMACEUTICAL FORMULATIONS

[0113] The pharmaceutical formulations of the present invention are useful, inter alia, for the treatment, prevention or improvement of any disease or disorder associated with PD-1 activity, including PD-1-mediated diseases or disorders. Non-limiting and illustrative diseases and disorders that can be treated or prevented by administration of the pharmaceutical formulations of the present invention include viral infections, autoimmune diseases and various cancers such as, for example, brain cancer, lung cancer, prostate cancer, colorectal cancer, head and neck cancer, skin cancer, various types of blood cancer and endometrial cancer. EXAMPLES

[0114] The following examples are presented so as to provide those skilled in the art with a complete description and disclosure of how to make and use the methods and compositions of the invention and are not intended to limit the scope of what the inventors consider their invention. Efforts have been made to ensure accuracy with respect to the numbers used (e.g., quantities, temperature, etc.), Petition 870220073975, dated 08 / 17 / 2022, page 60 / 245 50 / 130 However, some experimental errors and deviations should be considered. Unless otherwise indicated, parts are parts per mole, molecular weight is average molecular weight, temperature is in degrees Celsius, and pressure is equal to or close to atmospheric pressure. Example 1: Development of an Anti-PD-1 Antibody Formulation

[0115] The objectives of the formulation activities were to develop a formulation with the following attributes: • A liquid formulation with a concentration of anti-PD-1 antibody sufficient to administer a dose of 250 mg or more by intravenous infusion; • A near-iso-osmolar formulation that is stable after dilution with commonly used diluents, for example, 0.9% sodium chloride injection or 5% dextrose injection, for intravenous infusion; • A formulation that is compatible and stable in type 1 glass vials and standard serum stoppers as a package; • A sterile (SD) drug solution that supports long-term stability; A formulation that minimizes high molecular weight (HMW) antibody species when subjected to handling and thermal stress; A formulation that minimizes variations in the relative distribution of antibody-laden species when subjected to thermal stress; and a formulation that retains biological activity when subjected to handling and thermal stress.

[0116] During the development of the formulation, three primary protein stress conditions (representing extreme handling conditions beyond which the antibody drug would not be Petition 870220073975, dated 08 / 17 / 2022, page 61 / 245 51 / 130 submitted during handling, manufacturing, transport, storage, and labeling) were employed to develop and optimize antibody formulations and evaluate the effects of potential real-world stresses on drug stability. These stress conditions include: • Agitation (vortexing) of the protein solution at room temperature. The vortex in glass flasks exceeds the agitation that occurs during protein handling and manufacturing. • Incubation of the protein solution at an elevated temperature (37°C, 40°C or 45°C) relative to the proposed storage condition of DP (2°C-8°C). • Subjecting the protein to multiple freeze-thaw cycles. Since the protein will undergo at least one freeze-thaw cycle during DP manufacturing, multiple freeze-thaw cycles simulate and exceed the actual stress the protein is expected to experience.

[0117] There were four main objectives in the initial work of developing the formulation:

[0118] 1. Buffer and pH Selection: The choice of buffer and pH can have a large effect on protein stability; therefore, deciding on the ideal buffer and pH species is an important process. Studies are presented in these sections that demonstrate the rationale for choosing the ideal antibody buffer and pH.

[0119] 2. Selection of surfactant or organic cosolvent: An organic surfactant or cosolvent, such as polysorbate, is usually required to prevent protein precipitation or aggregation when agitated. Soluble protein may be subjected to agitation when handled, filtered, mixed, manufactured, transported, and administered. The antibody drug substance in a simple buffered solution may become visibly cloudy with excess Petition 870220073975, dated 08 / 17 / 2022, page 62 / 245 52 / 130 agitation. Therefore, it was determined that the stability of the protein during handling and agitation was important.

[0120] 3. Identification / selection of stabilizing / tonifying excipients: The addition of sugars, salts, and amino acids was examined for their ability to improve antibody stability to thermal stress and increase the shelf life of the DP. The rationale for including these thermal stabilizers, as well as studies identifying the ideal concentrations in the final formulation, are presented here.

[0121] 4. Selection of antibody concentration: The effect of antibody concentration on drug stability with the selected excipients was examined.

[0122] Initial formulation development activities were conducted using 5-50 mg / mL of anti-PD-1 antibody and involved screening organic cosolvents, thermal stabilizers, and buffers in liquid anti-PD-1 antibody formulations to identify excipients that are compatible with the protein and enhance its stability while maintaining near-physiological osmolarity and low viscosity for intravenous and subcutaneous injection. Buffering conditions were also examined to determine the optimal pH for maximum protein stability (described in Examples 4, 6, and 7).

[0123] The results of this initial formulation development work were used to develop a suitable initial formulation for Phase 1 clinical studies. The Phase 1 formulation also served as a reference for optimizing subsequent phase clinical and commercial formulations.

[0124] With the knowledge gained from the initial formulation development, the subsequent phase development activities involved optimizing the pH, surfactant concentration and Petition 870220073975, dated 08 / 17 / 2022, page 63 / 245 53 / 130 stabilizers to identify excipients that improve protein stability at low and high protein concentrations (up to 175 mg / mL of mAb1) (described in Examples 5, 8, 9 and 10).

[0125] Throughout formulation development, the formulations were evaluated for stress and storage stability. The methods used to evaluate stability in formulation development studies are described in Example 3 here. Examples 11 and 12 describe the storage and stress stability of the formulations.

[0126] Example 13 describes the stability of formulations when excipients varied within specific ranges.

[0127] The results generated from these studies were used to develop stable liquid formulations suitable for clinical use for intravenous (IV) or subcutaneous (SC) administration. Example 14 describes containers used for the formulations herein. Examples 15, 16, and 17 describe the compatibility and stability of the formulations in glass vials, pre-filled syringes, and intravenous administration devices. These formulations met the objectives defined for the formulation development: • The formulations developed are suitable for the dosages developed; • Iso-osmolar tonicity under physiological conditions; The osmolality of the 50 mg / mL formulation is approximately 318 mOsm / kg; • Sterile DP solution that supports long-term stability in the liquid state; Minimal formation of HMW antibody species occurs after prolonged storage at 2-8°C; Little or no change in the relative distribution of antibody-bearing species occurs after storage. Petition 870220073975, dated 08 / 17 / 2022, page 64 / 245 54 / 130 prolonged at 2-8°C; and minimal formation of subvisible particles was observed in DP under accelerated storage and stress conditions and after storage at 5°C for 12 months.

[0128] Other attributes of the formulations will become apparent from the description here.

[0129] Anti-PD-1 Antibodies: Anti-PD-1 antibodies are described in US20150203579, which is incorporated herein in its entirety. The exemplary antibody used in the Examples below is a fully human anti-PD-1 antibody H4H7798N (as described in US20150203579, known as REGN2810 or cemiplimab) comprising a heavy chain comprising the amino acid sequence SEQ ID NO: 9 and a light chain comprising the amino acid sequence SEQ ID NO: 10; an HCVR / LCVR amino acid sequence pair comprising SEQ ID NOs: 1 / 2; and heavy and light chain CDR sequences comprising SEQ ID NOs: 3-8; and referred to herein as mAb1. Example 2: Illustrative Formulations

[0130] In certain embodiments, mAb1 is formulated as an aqueous buffered formulation containing from 5 mg / mL ± 0.75 mg / mL to 250 mg / mL ± 45.0 mg / mL of mAb1, histidine buffer at 10 mM ± 2 mM, polysorbate at 0.2% ± 0.1% w / v, sucrose at 1% ± 0.2% to 10% ± 2% w / v, proline at 1% ± 0.02% to 5% ± 1% w / v at a pH of 6.0 ± 0.3.

[0131] Exemplary formulations include: • A stable, low-viscosity pharmaceutical formulation comprising: 25 mg / mL ± 3.75 mg / mL of mAb1, histidine buffer at 10 ± 2 mM, polysorbate 80 at 0.2% ± 0.1% w / v, sucrose at 5% ± 1% w / v, L-proline at 1.5% ± 0.3% w / v at a pH of 6.0 ± 0.3. • A stable, low-viscosity pharmaceutical formulation Petition 870220073975, dated 08 / 17 / 2022, page 65 / 245 55 / 130 comprising: 50 mg / mL ± 7.5 mg / mL mAbl, histidine buffer at 10 ± 2 mM, polysorbate 80 at 0.2% ± 0.1% w / v, sucrose at 5% ± 1% w / v, L-proline at 1.5% ± 0.3% w / v at a pH of 6.0 ± 0.3. • A stable, low-viscosity pharmaceutical formulation comprising: 150 ± 23 mg / mL of mAb1, histidine buffer at 10 ± 2 mM, polysorbate 80 at 0.2% ± 0.1% w / v, sucrose at 5% ± 1% w / v, L-proline at 1.5% ± 0.3% w / v at a pH of 6.0 ± 0.3. • A stable, low-viscosity pharmaceutical formulation comprising: 175 ± 27 mg / mL of mAb1, histidine buffer at 10 ± 2 mM, polysorbate 80 at 0.2% ± 0.1% w / v, sucrose at 5% ± 1% w / v, L-proline at 1.5% ± 0.3% w / v at a pH of 6.0 ± 0.3.

[0132] Example 3: Methods Used to Evaluate Formulation Stability

[0133] The following tests were applied to evaluate the stability of the formulation: • Color and appearance by visual inspection • pH • Turbidity measured by DO increase at 405nm or nephelometry • Particulate matter analysis performed by microflow imaging (MFI) (reported as particle count obtained as such) and light obscuration (HIAC) • Protein concentration by ultra-high performance reversed-phase liquid chromatography (RP-UPLC) • Purity by size exclusion ultra-high performance liquid chromatography (SE-UPLC) or reduced and non-reduced microchip capillary electrophoresis of sodium dodecyl sulfate (MCE-SDS) • Charge variant analysis by cation exchange chromatography-ultra-high performance liquid chromatography Petition 870220073975, dated 08 / 17 / 2022, page 66 / 245 56 / 130 (CEX-UPLC) or capillary isoelectric focusing imaging (iCIEF) • Potency via bioassay: The relative potency of each sample is determined using a bioassay and is defined as: (IC50 of the reference sample / IC50 of the sample) * 100%. The measured potency of the storage stability samples must be between 50% and 150% of the measured potency of the reference standard.

[0134] The physical stability of a formulation refers to properties such as color, appearance, pH, turbidity, and protein concentration. The presence of visible particulates in the solution can be detected by visual inspection. A solution passes visual inspection if it is clear to slightly opalescent, essentially free of visible particulates, and colorless to light yellow. In addition, turbidity, measured by OD at 405 nm, can also be used to detect particulates in solution. An increase in OD at 405 nm may indicate the presence of particulates, as may an increase in opalescence or color variation of the test articles. MFI is used to measure subvisible particulates with a size > 2 μm. The protein concentration of mAb1 is measured using an RP-UPLC assay and reported as a percentage of protein recovery relative to the raw material. In the RP-UPLC assay, mAb1 is eluted from the RP column as a single peak.Protein concentration is determined from the total peak area of ​​mAb1 by comparing it to a calibration curve generated using mAb1 standards. The percentage of recovery is calculated based on the measured protein concentration relative to the initial protein concentration.

[0135] Chemical stability refers to the formation of covalently modified forms (e.g., covalent aggregates, cleavage products, or charge variant forms) and non-covalently modified forms (e.g., non-covalent aggregates) of the protein. The higher and lower molecular weight degradation products Petition 870220073975, dated 08 / 17 / 2022, page 67 / 245 57 / 130 molecular weights can be separated from native mAb1 using SE-UPLC and MCE-SDS methods. The percentage of mAb1 degraded in the SE-UPLC and MCE-SDS methods is calculated from the ratio between the area of ​​all non-native peaks and the total area of ​​all mAb1 peaks. Variant charge forms of mAb1 are decomposed using CEX-UPLC and iCIEF. In the CEX-UPLC method, peaks with retention times earlier than the main peak are labeled as acidic peaks; peaks with retention times later than the main peak are labeled as basic peaks. In the iCIEF method, peaks focused at a pI lower than that of the main peak are labeled as acidic, while those focused at a pI higher than that of the main peak are labeled as basic. Example 4: Effect of Different Buffers and pH

[0136] The effect of buffer and pH on the thermal stability of mAb1 was examined in liquid formulations by incubating 5 mg / mL of mAb1 at 45 °C for 28 days in a series of buffer systems at different pH ranges. The following pH and buffer systems were studied: acetate (pH 4.5, 5.0, 5.5), histidine (pH 5.5, 6.0, 6.5), and phosphate (pH 6.0, 6.5, 7.0). Based on the results of SEUPLC analysis, maximum protein stability was observed when mAb1 was formulated at a pH between 6.0 and 6.5 in histidine buffer (Table 1). Petition 870220073975, dated 08 / 17 / 2022, page 68 / 245 Table 1: Effect of buffer and pH on the stability of 5 mg / mL mAbl incubated at 45°C for 28 days. Formulation 5 mg / mL mAbl, 10 mM buffer Filling volume 0.4 mL Container 2 mL borosilicate glass bottle type 1 with a FluroTec® coated butyl rubber stopper pH / buffer Color and appearance Turbidity (DO increase at 405 nm) % protein recovered by RPUPLC Variation in purity by SE-UPLC 1> Variation in charge variants by CEX-UPLC a> % HMW % Native % LMW % Acid % Main % Basic pH 4.5, acetate Approved 0.00 87 5.2 -7.2 2.0 11.1 -15.4 4.3 pH 5.0, acetate Approved 0.00 82 5.0 -5.9 0.9 7.5 -10.9 3.4 pH pH 5.5, acetate Approved 0.00 90 4.7 -5.4 0.7 12.8 -13.7 0.9 pH 5.5, histidine Approved 0.00 97 5.6 -6.4 0.8 10.8 -12.4 1.6 pH 6.0, histidine Approved 0.00 86 1.9 -2.4 0.6 13.4 -12.6 -0.7 pH 6.5, histidine Approved 0.00 84 1.2 -1.8 0.7 25.4 -21.3 -4.1 pH 6.0, phosphate Approved 0.01 92 3.7 -4.3 0.6 24.8 -21.8 -3.0 pH 6.5, phosphate Approved 0.03 91 4.9 -5.8 0.9 49.6 -40.3 -9.3 pH 7.0, phosphate Approved 0.03 95 10.4 -11.6 1.2 56.5 -42.2 -14.3 58 / 130

[0137] Reported as a relative variation in purity from the raw material. The raw material (without incubation) contains > 97.2% native peak by SE-UPLC and > 49.0% main peak by CEX-UPLC in all formulations. Petition 870220073975, dated 08 / 17 / 2022, page 69 / 245 59 / 130 CEX = cation exchange; DS = medicinal substance; HMW = high molecular weight; LMW = low molecular weight; OD = optical density; PR = reversed phase; SE = size exclusion; UPLC = high-performance liquid chromatography

[0138] Based on the results of the CEX-UPLC analysis, maximum protein stability was observed when mAb1 was formulated at a pH between 5.5 and 6.0 in histidine buffer or at a pH between 5.0 and 5.5 in acetate buffer. These analyses also revealed that aggregation (i.e., formation of HMW species), fragmentation (i.e., formation of LMW species), and charge variant formation were the main degradation pathways. Histidine buffer was selected as the formulation buffer because it provided the best overall level of protein stabilization with respect to the formation of HMW and LMW species and charge variant formation. A pH of 6.0 was chosen for the formulation because the formation of HMW species and charge variants, which are the main degradation pathways, were minimized at this pH. Based on these results, 10 mM histidine buffer at pH 6.0 was chosen for the formulation of mAb1. Example 5: pH Screening in Histidine Buffers

[0139] The effect of buffer and pH on the thermal stability of mAb1 was examined in high-concentration liquid formulations. mAb1 at 150 mg / mL was incubated at 45 °C for 28 days in a series of histidine buffers with pH ranging from 5.3, 5.5, 5.8, 6.0, and 6.3 with and without thermal stabilizers. With 9% sucrose, based on SE-UPLC analysis results, maximum protein stability was observed when mAb1 was formulated at a pH between 5.8 and 6.3 in histidine buffer (Figure 1). Based on CEX-UPLC analysis results, maximum protein stability was observed when mAb1 was formulated at a pH between 5.3 and 6.0 in histidine buffer (Table 2). Petition 870220073975, dated 08 / 17 / 2022, page 70 / 245 Table 2: Effect of pH on the stability of mAbl at 150 mg / mL incubated at 45°C for 28 days Formulation mAbl at 150 mg / mL, histidine at 10 mM Filling volume 0.4 mL Container / seal 2 mL borosilicate glass bottle type 1 with FluroTec® coated butyl rubber stopper pH / Stabilizer Color and appearance Turbidity (DO increase at 405 nm) % protein recovered by RP-UPLC Purity variation by SE-UPLC 1> Variation in charge variants by CEX-UPLC a) % HMW % Native % LMW % Acid % Main % Basic pH 5.3 / none Rejected 2> N / DN / DN / DN / DN / DN / DN / DN / D pH 5.5 / none Rejected b> N / DN / DN / DN / DN / DN / DN / DN / D pH 5.8 / none Rejected 0.40 95 46.2 -46.4 0.3 6.4 -10.2 3.9 pH 6.0 / none Rejected 0.51 99 41.2 -41.5 0.3 10.6 -7.3 -3.3 pH 6.3 / none Rejected 0.65 98 35.0 -35.4 0.4 19.6 -14.3 -5.3 pH 5.3 / 9% (w / v) Sucrose Approved 0.14 95 41.9 -42.1 0.3 7.9 -11.1 3.2 pH 5.5 / 9% (w / v) Sucrose Approved 0.16 99 30.8 -31.2 0.4 9.5 -12.5 2.9 pH 5.8 / 9% (w / v) Sucrose 9% (w / v) Approved 0.13 100 22.8 -23.3 0.5 9.7 -11.8 2.1 pH 6,0 / Sucrose at 9% (w / v) Approved 0.14 99 19.4 -19.9 0.5 13.5 -14.5 1.0 pH 6.3 / Sucrose at 9% (w / v) Approved 0.15 98 16.9 -17.4 0.5 22.4 -22.1 -0.4 60 / 130

[0140] Reported as a relative variation in purity in reaction to the raw material. The raw material (without incubation) contains > 94.0% native peak by SE-UPLC and > 48.7% main peak by CEX-UPLC in all formulations. bSample gelled. No other analysis was performed. NA = Not applicable Petition 870220073975, dated 08 / 17 / 2022, page 71 / 245 61 / 130 CEX, cation exchange; HMW, high molecular weight; LMW, low molecular weight; DO, optical density; PR, reversed phase; SE, size exclusion; UPLC, ultra-high performance liquid chromatography

[0141] These analyses also revealed that aggregation (i.e., formation of HMW species) and charge variant formation were the main degradation pathways. A pH of 6.0 was chosen for the DP formulation because the formation of HMW species and charge variants, which are the main degradation pathways, were minimized at this pH. Based on these results, 10 mM histidine buffer at pH 6.0 was chosen for the DP formulation with a high concentration of mAb1. Example 6: Selection of Protectors Against Stress from Agitation

[0142] Stabilizers, such as surfactants and organic cosolvents, are often added to antibody formulations to protect the protein against agitation-induced aggregation. The effect of cosolvents and organic surfactants on the agitation stress stability and thermal stability of mAb1 at 5 mg / mL was examined in liquid formulations. The following cosolvents and surfactants were evaluated: polysorbate 20 at 0.1%, polysorbate 80 at 0.1%, and PEG3350 at 1.0%. The results of the agitation stress stability studies are summarized in Table 3. Petition 870220073975, dated 08 / 17 / 2022, page 72 / 245 Table 3: Effect of organic cosolvents and surfactants on the stability of mAbl at 5 mg / mL after agitation (120 minutes of vortexing) Formulation mAbl at 5 mg / mL, histidine at 10 mM, pH 6.0 Filling volume 0.4 mL Container 2 mL borosilicate glass bottle type 1 with a FluroTec® coated butyl rubber stopper 4432 / 50 Cosolvent / Surfactant Color and appearance Turbidity (OD increase at 405 nm) pH % protein recovered by RP-UPLC Variation in Purity by SE-UPLC 1> Variation in charge variants by CEX-UPLC at % HMW % Native % LMW % Acid % Principal % Basic Without cosolvent / surfactant Rejected 1.69 6.0 76 15.3 -23.7 -8.4 -2.3 -1.7 3.9 Sucrose at 5% (w / v) Rejected 1.75 6.0 64 9.4 -12.8 3.4 -1.7 -0.1 1.8 Polysorbate 20 at 0.1% (w / v) b> Approved 0.00 6.0 102 -0.1 -0.8 0.8 0.2 0.2 -0.3 Polysorbate 80 at 0.1% (w / v) b> Approved 0.00 6.0 100 -0.2 -0.5 0.4 0.2 -0.1 -0.1 PEG3350 at 1% (w / v) b> Rejected 0.20 6.0 93 0.3 -0.5 0.2 -0.2 -0.3 0.4 62 / 130

[0143] aReported as a relative variation in purity compared to the raw material. The raw material (without incubation) contains > 98.2% native peak by SE-UPLC and > 49.1% main peak by CEX-UPLC in all 5 formulations.bThe formulation also contains 5% sucrose. Petition 870220073975, dated 08 / 17 / 2022, page 73 / 245 63 / 130 CEX = cation exchange; HMW = high molecular weight; LMW = low molecular weight; OD = optical density; PR = reversed phase; SE = size exclusion; UPLC = high-performance liquid chromatography

[0144] mAb1 was unstable when vortexed for 120 min in the absence of an organic cosolvent or surfactant. After vortexing in the absence of a cosolvent or surfactant, the solution became cloudy, exhibited a substantial increase in turbidity, and had a 15.3% increase in aggregates, as determined by SE-UPLC, in addition to a 24% loss in protein recovery by RP-UPLC (Table 3). 1% PEG3350 did not provide sufficient stabilization of mAb1 after 120 min of vortexing. In the presence of 1% PEG3350, the solution became cloudy and exhibited an increase in turbidity (Table 3). In contrast, 0.1% polysorbate 20 and 0.1% polysorbate 80 protected mAb1 against vortex-induced instability to the same extent (Table 3).

[0145] However, the formulation containing 0.1% polysorbate 80 exhibited a decrease in the amount of aggregates compared with the formulation containing 0.1% polysorbate 20 when incubated at 45 °C (Table 4).Polysorbate 80 at 0.1% was chosen as the surfactant for the DP mAb1 formulation because it stabilized the protein under agitation stress, had less of a negative effect on the thermal stability of the protein than polysorbate 20 (as determined by SE-UPLC and CEX-UPLC analyses), and has a safe history of use in monoclonal antibody formulations.

[0146] Example 7: Selection of Heat Stress Protectors

[0147] Stabilizers, such as sucrose, are frequently added to antibody formulations to increase the thermal stability of the protein in liquid formulations. mAb1 at five (5) mg / mL in a liquid formulation exhibited enhanced stability when formulated with 5% sucrose and incubated under accelerated conditions (Table 4). Petition 870220073975, dated 08 / 17 / 2022, page 74 / 245 Table 4: Effect of organic cosolvents and surfactants on the stability of mAbl at 5 mg / mL incubated at 45°C for 29 days. Formulation mAbl at 5 mg / mL, histidine at 10 mM, pH 6.0 Filling volume 0.4 mL Container / seal 2 mL borosilicate glass bottle type 1 with FluroTec® coated butyl rubber stopper 4432 / 50 Co-solvent / surfactant Color and appearance Turbidity (increase in OD at 405 nm) pH % protein recovered by RP-UPLC Change in Purity by SE-UPLC 1> Change in charge variants by CEX-UPLC a) % HMW % Native % LMW % Acid % Principal % Basic Without co-solvent / surfactant Approved 0.00 6.1 96 2.8 -3.2 0.5 10.8 -10.7 -0.2 Sucrose at 5% (w / v) Approved 0.01 6.1 99 1.6 -2.0 0.3 12.6 -11.7 -0.9 Polysorbate 20 at 0.1% (w / v) b> Approved 0.00 6.0 92 27.6 -28.4 0.7 0.1 -24.0 23.9 Polysorbate 80 at 0.1% (w / v) Approved 0.00 6.1 96 12.8 -14.1 0.9 4.4 -20.4 15.9 PEG3350 at 1% (w / v) Approved 0.00 6.1 90 8.4 -8.0 -0.4 0.4 -25.0 24.5 64 / 130

[0148] aReported as a reactive variation in purity relative to the raw material. The raw material (without incubation) contains > 98.2% native peak by SE-UPLC and > 49.1% main peak by CEX-UPLC in all 5 formulations.bThe formulation also contains 5% sucrose. Petition 870220073975, dated 08 / 17 / 2022, page 75 / 245 65 / 130 CEX = cation exchange; HMW = high molecular weight; LMW = low molecular weight; OD = optical density; PR = reversed phase; SE = size exclusion; UPLC = high-performance liquid chromatography

[0149] After incubation at 45°C for 29 days, the relative amount of HMW species increased from 1.7% in the formulation containing 5% sucrose compared to an increase of 2.8% in the control formulation without sucrose. For this reason, sucrose was chosen as a thermal stabilizer. To make the formulation isotonic and maximize thermal stability, the sucrose concentration was increased to 10% for the mAb1 formulation. Example 8: Stabilizer Optimization

[0150] The objective of optimizing thermal stabilizers was to identify stabilizing components that could be used to develop a DP formulation that supports an antibody concentration of up to 200 mg / mL. 10% sucrose was selected in the initial formulation. It was observed that, with 10% sucrose, the viscosity of mAb1 was approximately 20 cP at 20°C and was considered too high for a robust, later-stage commercial product. Therefore, a modified mAb1 formulation was needed that exhibited both favorable stability and lower viscosity.

[0151] Sucrose was chosen as a thermal stabilizer for mAb1 during the development of the low-concentration formulation. For the development of the high-concentration formulation, different concentrations of sucrose and L-proline were evaluated for the stability and viscosity of mAb1 at concentrations of 150 and 175 mg / mL at 25°C (Table 5) and at 40°C for 1 month. The formation of HMW species decreased with increasing sucrose concentrations when the formulations were incubated at 40°C for 28 days. L-proline at 3% conferred similar stabilization to sucrose at 5% and Petition 870220073975, dated 08 / 17 / 2022, page 76 / 245 66 / 130 maximum stabilization was observed with 9% sucrose.

[0152] Although 9% sucrose provided slightly better stabilization compared to 3% L-proline, it also increased the viscosity of the formulation. At 175 mg / mL, the mAb1 formulation with 9% sucrose has a viscosity of 27 centipoise, which presents manufacturing and administration challenges. The 175 mg / mL mAb1 formulation with 3% proline has a viscosity of approximately 20 centipoise, which is manageable with the current manufacturing process. Petition 870220073975, dated 08 / 17 / 2022, page 77 / 245 Table 5: Accelerated stability of mAbl at high concentration with thermal stabilizers at 250°C for 1 month. Formulation mAbl at 210 mg / mL, histidine at 10 mM, pH 6.0 Filling volume 0.8 mL Container / seal 5 mL polycarbonate bottle with silicone-coated polypropylene screw cap Ex dpi entes Color and appearance Turbidity (DO increase at 405 nm) pH % mAbl recovered by RP-UPLC Purity by SE-UPLC Load variants by CEX-UPLC % HMW % Native % LMW % Acid % Main % Basic Raw material 1) Approved 0.00 6.0 100 2.9 96.6 0.5 25.3 47.4 27.3 Proline at 3% Approved 0.01 6.0 106 4.1 95.3 0.6 25.2 47.1 27.8 Sucrose at 3% Approved 0.00 6.0 107 4.7 94.8 0.6 25.2 46.5 28.3 Sucrose at 5% Approved 0.00 6.0 104 4.7 94.8 0.6 25.1 46.1 28.8 SDS / DP Formulation: mAbl at 150 mg / mL or 175 mg / mL, histidine at 10 mM, pH 6.0, PS 80 at 0.1% Filling Volume 0.4 mL Container / Seal 2 mL borosilicate glass bottle type 1 with FluroTec® coated butyl rubber stopper Raw Material 1) Approved 0.00 6.1 100 2.3 97.4 0.4 24.9 50.2 24.9 Sucrose at 9%, PS 80 at 0.1% Approved 0.00 6.1 101 3.7 95.7 0.7 28.4 47.7 23.9 Proline at 3%, PS 80 at 0.1% Approved 0.00 6.1 101 3.7 95.7 0.7 26.5 47.7 25.8 67 / 130

[0153] aThe pH, SE-UPLC, and CEX-UPLC results for 'Raw Material' are the average values ​​of the initial formulations. Petition 870220073975, dated 08 / 17 / 2022, page 78 / 245 68 / 130

[0154] However, based on storage stability at -20°C, -30°C and -80°C, it was found that the formulation with 3% proline was not as stable as the formulations with sucrose (Figure 2). As shown in Figure 2, formulation F3 (with 5% sucrose) was stable at -20°C, -30°C and -80°C with < 3% of HMW species.

[0155] The effect of L-proline on mAbl stabilization was examined with the 50 mg / mL formulation. After incubation at 45°C for 28 days, the formulation with L-proline showed lower levels of HMW species compared to the formulation without L-proline, showing that L-proline stabilizes the antibody at a concentration of 50 mg / mL (Table 6). Furthermore, the impact of L-proline on the antibody protein structure was examined using biophysical techniques (Fourier transform infrared spectroscopy, CD spectroscopy, fluorescence emission spectroscopy, and differential scanning calorimetry). The results showed that L-proline did not disrupt the secondary and tertiary structure of the antibody. Petition 870220073975, dated 08 / 17 / 2022, page 79 / 245 Table 6: Effect of stabilizers on the stability of mAbl at 50 mg / mL after incubation at 45°C for 28 days. Formulation mAbl at 50 mg / mL, L-histidine at 10 mM, sucrose at 5%, pH 6.0, polysorbate 80 at 0.2% Filling volume 0.6 mL Container / seal 2 mL type 1 glass bottle with FluroTec® coated chlorobutyl stopper 4432 / 50 Stabilizer (% w / v) Color and appearance Turbidity (OD increase at 405 nm) pH % protein recovered by RP-UPLC Variation in purity by SE-UPLC Variation in charge variants by CEX-UPLC % HMW % monomers % LMW % Acid % Principal % Basic Approved 0.02 6.0 96 12.1 -12.7 0.6 10.8 -11.9 1.0 L-proline a 1.5% Approved 0.02 6.1 96 11.3 -11.9 0.6 11.0 -11.8 0.8 L-proline at 3.0% Approved 0.01 6.0 96 10.9 -11.5 0.6 12.8 -12.9 0.1 69 / 130

[0156] a) Reported as a change in purity compared to the raw material. The raw material (without incubation) contains > 98.5% monomer peak by SE-UPLC and > 52.8% main peak by CEX-UPLC in all three formulations. Petition 870220073975, dated 08 / 17 / 2022, page 80 / 245 70 / 130

[0157] CEX, cation exchange; DS, medicinal substance; HMW, high molecular weight; LMW, low molecular weight; OD, optical density; PR, reversed phase; SE, size exclusion; UPLC, ultra-high performance liquid chromatography

[0158] The effect of different stabilizers on the thermal stability of high concentrations (150 and 175 mg / mL) of mAb1 was further examined in liquid formulations. The stabilizers evaluated were 9% (w / v) sucrose, 3% (w / v) L-proline, and 5% (w / v) sucrose with 1.5% (w / v) L-proline. The results of the accelerated stability study are summarized in Table 7. Petition 870220073975, dated 08 / 17 / 2022, page 81 / 245 Table 7: Effect of stabilizers on mAbl stability after incubation at 25°C and 40°C Formulation mAbl at 150 or 175 mg / mL, histidine at 10 mM, pH 6.0, polysorbate 80 at 0.1% Filling volume 0.5 mL Container / Seal 2 mL borosilicate glass bottle type 1 with FluroTec® coated butyl rubber stopper 4432 / 50 Incubation 25°C for 3 months Stabilizer (% w / v) mAbl Cone. (mg / mL) Color and appearance Turbidity (increase in DO at 405 nm) pH % of protein recovered by RP-UPLC Variation in purity by SE-UPLC 1> Variation in charge variants by CEX-UPLC % HMW % Native % LMW % Acid % Main % Basic Sucrose at 9% 150 Approved 0.02 6.2 110 2.6 -2.8 0.2 6.0 -4.2 -1.9 L-proline at 3% 175 Approved 0.00 6.2 112 2.4 -2.5 0.2 6.5 -4.1 -2.4 Sucrose at 5%, L-proline at 1.5% 175 Approved 0.00 6.2 110 2.3 -2.6 0.2 6.0 -3.7 -2.4 Incubation at 40°C for 28 days Stabilizer (% w / v) mAbl Cone.(mg / mL) Color and appearance Turbidity (increase in DO at 405 nm) pH % of protein recovered by RP-UPLC Variation in purity by SE-UPLC a> Variation in charge variants by CEX-UPLC a % HMW % Native % LMW % Acid % Main % Basic Sucrose at 9% 150 Approved 0.04 6.1 102 6.9 -7.5 0.5 9.4 -9.7 0.3 3% L-proline 175 Approved 0.03 6.2 103 11.7 -12.3 0.6 12.1 -10.7 -1.4 Sucrose at 5%, L-proline at 1.5% 175 Approved 0.03 6.1 103 8.8 -9.4 0.7 11.4 -10.2 -1.2 71 / 130

[0159] Reported as a change in purity compared to the raw material. The raw material (without incubation) contains > 97.3% native peak by SE-UPLC and > 49.6% main peak by CEX-UPLC in all three formulations. Petition 870220073975, dated 08 / 17 / 2022, page 82 / 245 72 / 130 CEX, cation exchange; HMW, high molecular weight; LMW, low molecular weight; OD, optical density; PR, reversed phase; SE, size exclusion; UPLC, ultra-high performance liquid chromatography

[0160] After incubation at 40°C for 28 days, 9% sucrose provided the best stabilization and had the highest viscosity among the high-concentration formulations. The 5% sucrose / 1.5% L-proline formulation ranked second in terms of stability after 28 days at 40°C. After incubation at 25°C for three months, the stability of mAb1 was almost the same in all formulations examined; however, the formulation with 5% sucrose / 1.5% L-proline was slightly better than the other two formulations. However, after incubation at -20°C, -30°C, and -80°C, 5% and 9% sucrose provided better stability than 3% proline (Figure 3). mAb1 at 175 mg / mL with 5% sucrose / 1.5% L-proline has a viscosity of 14 cP at 20°C. To provide a formulation that produces an isotonic solution and achieves a better balance between stability and viscosity, 5% sucrose / 1.5% L-proline was selected for the development of a post-phase PD formulation. Example 9: Selection of Viscosity Modifiers

[0161] The viscosity of protein formulations increases exponentially as the protein concentration increases. When the viscosity begins to exceed approximately 10 to 15 cP at 20°C, the formulation viscosity must be taken into account in formulation development: this is simply because viscosity correlates with ease of injection via a pre-filled syringe (PFS) or other needle-based delivery device; most importantly, maintaining a reasonably low viscosity is fundamental to the development of a delivery device, such as an autoinjector. The effect of Petition 870220073975, dated 08 / 17 / 2022, p. 83 / 245 The viscosity of formulations with 73 / 130 excipients was examined in a liquid formulation with the following potential viscosity modifiers: proline, arginine.HCl, histidine.HCl, magnesium acetate, and NaCl. Figure 4 summarizes the viscosity of mAb1 at 150 mg / mL with the viscosity modifiers. Arginine.HCl, histidine.HCl, magnesium acetate, and NaCl at 25-100 mM decrease the viscosity of mAb1 formulations at 150 mg / mL.

[0162] The impact of viscosity modifiers on the stability of the mAb1 formulation was also examined. Formulations of mAb1 at 150 mg / mL with viscosity modifiers, such as arginine.HCl, histidine.HCl, magnesium acetate, and NaCl, were prepared and incubated at 45 °C for 28 days. The results are shown in Figure 5. It was observed that maximum protein stability was observed when mAb1 was formulated without the viscosity modifiers; all these viscosity-modifying salts negatively affect the stability of mAb1. Therefore, salts were not included in the final formulation.

[0163] L-proline, as a stabilizer, minimized the viscosity of the solution for antibody concentrations equal to or greater than 50 mg / mL. The results of accelerated stability studies for high-concentration antibodies with varying amounts of L-proline, with and without sucrose, are summarized in Table 7. After incubation at 25°C for three months, the formulation with 5% sucrose / 1.5% L-proline provided slightly better stability compared to the other two formulations with respect to the formation of HMW species. After incubation at 40°C for 28 days, the formulation containing 9% sucrose provided better stabilization, and the formulation with 5% sucrose / 1.5% L-proline came in second. The formulation with 9% sucrose has a viscosity of 20 cP at 175 mg / mL of antibody, while the viscosity of Petition 870220073975, dated 08 / 17 / 2022, page 84 / 245 A formulation at 175 mg / mL with 5% sucrose / 1.5% L-proline has a viscosity of 14 cP at 20°C. The addition of L-proline to the formulation was important to decrease viscosity at high protein concentrations, as well as to stabilize the antibody.

[0164] In summary, 5% sucrose / 1.5% L-proline was selected for both 50 mg / mL antibody formulations and high-concentration antibodies. This combination of excipients achieved an isotonic formulation with acceptable stability and viscosity at all antibody concentrations tested (up to 175 mg / mL). Example 10: Optimization of Polysorbate (PS) Concentration

[0165] During formulation development, the formation of higher molecular weight species and an increase in turbidity were observed when the mAb1 formulation was agitated without surfactant. The protein was stabilized by agitation through the addition of polysorbate 80 (PS 80). During the development of the high-concentration liquid mAb1 formulation, instability upon agitation was observed as an increase in high molecular weight species. A study was conducted to determine the minimum amount of polysorbate 80 required to protect mAb1 up to 175 mg / mL against agitation-induced instability. The formulations in this study contained 5% sucrose and 1.5% L-proline so that the effect of polysorbate 80 could be studied with a formulation composition more representative of the final formulation. The nominal concentrations of polysorbate 80 included in the study were 0%, 0.02%, 0.04%, 0.06%, 0.08%, 0.1%, 0.15%, and 0.2% (w / v).In the absence of polysorbate 80, the solution became cloudy and exhibited a significant increase in turbidity after vortex agitation. A concentration-dependent reduction in the % HMW was observed after 120 minutes of agitation. A concentration of 0.15-0.2% polysorbate 80 was found to be sufficient to stabilize mAb1 at 150 mg / mL. Petition 870220073975, dated 08 / 17 / 2022, page 85 / 245 75 / 130 and 175 mg / mL against agitation-induced aggregation (Table 8). The addition of 0.2% (w / v) polysorbate 80 (nominal value) completely prevented the formation of HMW species after agitation for 120 minutes. Petition 870220073975, dated 08 / 17 / 2022, page 86 / 245 Table 8: Stability of mAbl at 150 mg / mL and 175 mg / mL with PS 80 after 120 minutes of agitation. Filling volume 0.4 mL Container / Seal 2 mL borosilicate glass bottle type 1 with a FluroTec® coated butyl rubber stopper Formulation [PS 80] Color and appearance Turbidity (OD increase at 405 nm) pH % mAbl recovered by RP-UPLC % HMW increase by SE-UPLC mAbl at 150 mg / mL, Histidine at 10 mM, pH 6.0 sucrose at 9% 0.00 Rejected 0.38 6.0 Not applicable 0.02 Approved 0.00 6.0 97 4.7 0.04 Approved 0.00 6.0 100 0.3 0.06 Approved 0.00 6.0 102 0.3 0.08 Approved 0.04 6.0 99 0.0 0.10 Approved 0.01 6.0 101 0.1 0.15 Approved 0.00 6.0 98 0.1 0.20 Approved 0.00 6.0 106 0.1 mAbl at 175 mg / mL, Histidine at 10 mM, pH 6.0, Proline at 3% 0.00 Rejected 0.38 6.0 Not applicable 0.02 Approved 0.06 6.1 100 7.0 0.04 Approved 0.00 6.0 97 2.4 0.06 Approved 0.01 6.1 700 2.3 0.08 Approved 0.03 6.1 98 0.9 0.10 Approved 0.00 6.1 102 0.4 0.15 Approved 0.00 6.1 102 0.1 76 / 130 Petition 870220073975, dated 08 / 17 / 2022, page 87 / 245 Filling volume 0.4 mL Container / Seal 2 mL borosilicate glass bottle type 1 with a FluroTec® coated butyl rubber stopper Formulation [PS 80] Color and appearance Turbidity (OD increase at 405 nm) pH % mAbl recovered by RP-UPLC % increase in HMW by SE-UPLC 0.20 Approved 0.00 6.1 101 0.1 mAbl at 175 mg / mL, Histidine at 10 mM, pH 6.0, sucrose at 5% Proline at 1.5% 0.00 Rejected 0.36 6.1 Not applicable 0.02 Approved 0.01 6.1 97 3.5 0.04 Approved 0.01 6.1 98 2.0 0.06 Approved 0.01 6.1 100 1.3 0.08 Approved 0.01 6.1 99 1.0 0.10 Approved 0.01 6.0 102 0.3 0.15 Approved 0.00 6.1 101 0.2 0.20 Approved 0.00 6.1 98 0.0

[0166] Table 9 details the effect of polysorbate 80 concentration on the stability of mAbl at 175 mg / mL after agitation (120 minutes of vortexing). 77 / 130 Petition 870220073975, dated 08 / 17 / 2022, page 88 / 245 Table 9: Effect of polysorbate 80 concentration on the stability of mAbl at 175 mg / mL after agitation (120 minutes of vortexing) Formulation mAbl at 175 mg / mL, histidine at 10 mM, pH 6.0, sucrose at 5% (w / v), L-proline at 1.5% (w / v) Filling volume 0.4 mL Container / seal 2 mL borosilicate glass bottle type 1 with a FluroTec® PS 80 Cone coated butyl rubber stopper. Nominal (% w / v) Color and appearance Turbidity (DO increase at 405 nm) pH % of protein recovered by RPUPLC Variation in Purity by SE-UPLC 1> Variation in charge variants by CEX-UPLC a) % HMW % Native % LMW % Acid % Principal % Basic 0.00 % Rejected 0.36 6.0 N / DN / DN / DN / DN / DN / DN / D 0.02 % Approved 0.01 6.0 97 3.5 -3.5 0.0 -0.6 0.8 -0.1 0.04 % Approved 0.01 6.0 98 2.0 -2.0 0.0 0.1 -0.2 -0.2 0.06 % Approved 0.01 6.0 100 1.3 -1.3 0.0 -0.2 0.3 -0.1 0.08 % Approved 0.01 6.0 99 1.0 -1.0 0.0 -0.2 0.0 0.0 0.10 % Approved 0.02 6.0 102 0.3 -0.3 0.0 0.2 0.0 0.0 0.15 % Approved 0.00 6.1 101 0.2 -0.2 0.0 0.2 -0.3 0.1 0.20 % Approved 0.00 6.1 98 0.0 0.0 0.0 0.0 -0.1 0.2 78 / 130

[0167] Reported as a relative variation in purity compared to the raw material. The raw material (without incubation) contains > 97.4% native peak by SE-UPLC and > 48.2% main peak by CEX-UPLC in all formulations. Petition 870220073975, dated 08 / 17 / 2022, page 89 / 245 79 / 130 CEX, cation exchange; HMW, high molecular weight; LMW, low molecular weight; NA, not available; OD, optical density; PR, reversed phase; SE, size exclusion; UPLC, high-performance liquid chromatography

[0168] The ability of 0.2% (w / v) polysorbate 80 to protect mAb1 against agitation-induced instability was confirmed by another study with the final formulation at 50 mg / mL (Table 10). Petition 870220073975, dated 08 / 17 / 2022, page 90 / 245 Table 10: Effect of PS80 concentration on mAbl stability at 50 mg / mL after agitation (120 minutes of vortexing) Formulation mAbl at 50 mg / mL, L-histidine at 10 mM, pH 6.0, sucrose at 5% (w / v), L-proline at 1.5% (w / v) Filling volume 0.6 mL Container / seal 2 mL type 1 glass bottle with a FluorTec® Polysorbate 80 coated chlorobutyl stopper. (% w / v) Color and appearance Turbidity (increase in OD at 405 nm) pH % of protein recovered by RP-UPLC Variation in Purity by SE-UPLC to Variation in charge variants by CEX-UPLC to % HMW % monomers % LMW % Acid % Principal % Basic 0.0 % Approved 0.01 6.1 99 8.0 -8.0 0.0 1.7 -1.8 -0.3 0.2 % Approved 0.00 6.1 99 0.0 0.1 -0.1 -0.2 0.1 -0.2

[0169] a Reported as a relative variation in purity in relation to the raw material. The raw material (without 80 / 130 incubation) contains > 98.5% monomer peak by SE-UPLC and > 52.8% main peak by CEX-UPLC in all five formulations. Petition 870220073975, dated 08 / 17 / 2022, page 91 / 245 81 / 130 CEX, cation exchange; DS, medicinal substance; HMW, high molecular weight; LMW, low molecular weight; NA, not available; OD, optical density; PR, reversed phase; SE, size exclusion; UPLC, ultra-high performance liquid chromatography

[0170] Based on these results, 0.2% (w / v) polysorbate 80 was selected as the surfactant, as it provided sufficient stabilization to prevent the formation of HMW species under agitation stress.

[0171] Example 11: Storage and Stress Stability of Exemplary Formulations

[0172] The storage stability of mAb1a formulations at 50 mg / mL and 175 mg / mL in glass vials is shown in Table 11 and Table 12, and the accelerated stability and stress data for the two formulations are shown in Table 13 and Table 14, respectively. Stability research studies demonstrated that the mAb1a formulations at 50 mg / mL and 175 mg / mL in glass vials are stable for at least 24 months when stored at 2°C to 8°C. Furthermore, the mAb1a formulation at 50 mg / mL also exhibited excellent stability under accelerated and stress conditions. The formulation is stable when stored at 25°C for at least 3 months and at 40°C for at least 7 days, demonstrating the compatibility of the 50 mg / mL formulation with the main sealing components of the container.No appreciable changes were observed in color or appearance, turbidity, particulate matter, pH, protein concentration, purity measured by SE-UPLC or CEX-UPLC and iCIEF, and potency was maintained under these conditions. Petition 870220073975, dated 08 / 17 / 2022, page 92 / 245 Table 11: Stability test of the formulation at 50 mg / mL stored at 2-8°C Formulation: mAb1 at 50 mg / mL, L-histidine at 10 mM, sucrose at 5% (w / v), L-proline at 1.5% (w / v) and polysorbate 80 at 0.2% (w / v), pH 6.0. Filling volume: 1.2 mL. Container / Seal: 3 mL type 1 glass vials with a 13 mm FluroTec® coated chlorobutyl 4432 / 50 stopper. Assay: Storage duration at 2-8°C (months): 0 1 3 6 9 12 18 24 36. Color and appearance: Approved. Turbidity (DO increase at 405 nm): 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 Turbidity (NTU by nephelometry) 7.41 6.01 6.15 6.61 6.03 6.48 6.29 5.83 pH 6.0 6.0 6.0 6.1 6.0 6.0 6.0 6.0 Subvisible particulate matter analysis by HIAC (N / mL) > 10 pm 2 NR 21 8 NR 11 15 21 > 25 pm 0 NR 2 0 0 NR 1 1 1 Subvisible particulate matter analysis by IMF (N / mL) 2 to 10 pm 248 NR 887 1875 NR 607 125 4 776 > 10 pm 15 NR 26 44 NR 15 19 8 > 25 pm 4 NR 3 17 NR 1 3 1 % of protein recovered by RP-UPLC 100 100 98 103 101 105 98 98 Purity by MCE-SDS Not reduced;% peak 99.2 NR NR 98.7 NR 98.9 99.2 99.2; 82 / 130 Petition 870220073975, dated 08 / 17 / 2022, page 93 / 245 Main Reduced; % heavy + light chain 100 NR NR 1 00 NR 99.6 99.8 99.9 Purity by SE-UPLC % HMW 0.5 0.3 0.4 0.4 0.4 0.5 0.5 0.5 % monomers 99.2 99.1 99.2 99.2 99.1 99.2 99.2 99.1 % LMW 0.4 0.5 0.4 0.4 0.5 0.3 0.4 0.4 Charge variant analysis by CEXUPLC % Acid 18.9 19.0 18.7 19.1 19.4 19.1 20.7 20.5 % Main 53.9 53.5 54.5 53.7 53.6 55.8 53.8 53.4 % Basic 27.3 27.6 26.8 27.2 27.1 25.1 25.5 26.0 Analysis of charge variants by iCIEF % Acid 31.9 NR NR 32.3 NR 33.9 33.1 34.0 % Principal 54.7 NR NR 54.2 NR 54.0 54.0 53.9 % Basic 13.5 NR NR 13.5 NR 12.1 12.9 12.1 % relative potency (bioassay) 126 NR NR 127 NR 125 110 104 83 / 130 CEX, cation exchange; DS, medicinal substance; HMW, high molecular weight; iCIEF, capillary focusing isoelectric imaging; LMW, low molecular weight; IMF, microflow; monomer, intact antibody; NR, not required; OD, optical density; PR, reversed phase; SE, size exclusion; UPLC, high-performance liquid chromatography Petition 870220073975, dated 08 / 17 / 2022, page 94 / 245 Table 12: Stability test of the formulation at 175 mg / mL stored at 2-8°C Formulation: mAb1 at 175 mg / mL, L-histidine at 10 mM, sucrose at 5% (w / v), L-proline at 1.5% (w / v) and polysorbate 80 at 0.2% (w / v), pH 6.0. Filling volume: 1.2 mL. Container / Seal: 3 mL type 1 glass vials with a 13 mm FluroTec® coated chlorobutyl 4432 / 50 stopper. Assay: Storage duration at 5°C (months): 0 1 3 6 9 12 18 24 36. Color and appearance: Approved. Turbidity (DO increase at 405 nm): 0.00 0.01 0.00 0.00 0.00 0.00 0.00 0.00 Turbidity (NTU by nephelometry) 6.72 6.95 6.57 6.54 6.62 7.01 6.67 6.87 pH 6.0 6.0 6.0 6.0 6.0 6.0 6.0 6.1 Subvisible particulate matter analysis by HIAC (N / mL) > 10 pm 10 NR 28, 131 NR 30 35 55 > 25 pm 0 NR 11 56 NR 1 2 3 Subvisible particulate matter analysis by IMF (N / mL) 2 to 10 pm 144 NR 441 244 NR 265 319 117 > 10 pm 22 NR 36, 22 NR 29 28 8 > 25 pm 1 NR 11 7 NR 3 10 1 % of protein recovered by RP-UPLC 100 95 97 98 97 99 104 101 Purity by MCE-SDS Not reduced;% 97.9 NR 98.1 98.0 NR 97.9 98.0 97.9; 84 / 130 Petition 870220073975, dated 08 / 17 / 2022, page 95 / 245 Main peak reduced; % Heavy + Light Chain 100 NR 99.6 100 NR 99.8 99.8 99.8 Purity by SE-UPLC % HMW 0.6 0.6 0.7 0.7 0.8 0.9 1.0 1.0 % Monomer 99.1 98.9 99.1 98.7 98.7 98.7 98.6 98.6 % LMW 0.4 0.5 0.3 0.5 0.5 0.5 0.4 0.4 Charge Variant Analysis by CEXUPLC % Acid 18.7 18.3 18.1 18.7 19.1 19.0 19.0 20.3 % Principal 53.6 55.0 54.6 53.3 53.6 55.5 54.3 53.8 % Basic 27.8 26.7 27.4 28.1 27.3 25.5 26.8 25.9 Analysis of charge variants by iCIEF % Acid 31.8 NR 32.4 31.9 NR 33.9 32.3 32.0 % Principal 54.6 NR 54.6 54.9 NR 54.8 54.6 54.3 % Basic 13.6 NR 13.1 13.2 NR 11.3 13.1 13.7 % relative potency (bioassay) 102 NR NR 118 NR 110 91 107 85 / 130 CEX, cation exchange; DS, medicinal substance; HMW, high molecular weight; iCIEF, capillary focusing isoelectric imaging; LMW, low molecular weight; IMF, microflow; monomer, intact antibody; NR, not required; OD, optical density; PR, reversed phase; SE, size exclusion; UPLC, high-performance liquid chromatography Petition 870220073975, dated 08 / 17 / 2022, page 96 / 245 Table 13: Stability test of the 50 mg / mL formulation stored under accelerated and stress conditions. Formulation mAb1 at 50 mg / mL, L-histidine at 10 mM, sucrose at 5% (w / v), L-proline at 1.5% (w / v), polysorbate 80 at 0.2% (w / v), pH 6.0 Filling volume 1.2 mL Container / Seal 3 mL Type 1 glass vials with a 13 mm FluroTec® coated chlorobutyl 4432 / 50 stopper Storage at 25°C / 60% RH (months) Storage at 40°C (days) Assay 0 1 3 6 7 14 28 Color and appearance Approved Approved Approved Approved Approved Approved Approved Turbidity (DO increase at 405 nm) 0.00 0.00 0.00 0.00 0.00 0.00 0.00 Turbidity (NTU by Nephelometry) 7.41 6.05 6.54 6.90 6.10 6.60 6.93 pH 6.0 6.0 6.0 6.1 6.0 6.0 6.0 Subvisible particulate matter analysis by HIAC (# / mL) > 10 pm 2 NR 17 21 NR NR 18 > 25 pm 00 NR 00 1 NR NR 1 Subvisible particulate matter analysis by MFI (# / mL) 2-10 pm 248 NR 1618 1531 NR NR 1543 > 10 pm 15 NR 29 47 NR NR 41 > 25 pm 4 NR 2 15 NR NR 15 Percentage of protein recovered by RP-UPLC: 100, 100, 100, 103, 103, 102, 98. Purity by MCE-SDS: Not reduced.% Main Peak 99.2 NR 99.1 98.8 NR NR 98.9 Reduced; % 100 NR 99.5 100 NR NR 99.5; 86 / 130 Petition 870220073975, dated 08 / 17 / 2022, page 97 / 245 Heavy + Light Chain Purity by SE-UPLC % HMW 0.5 0.4 0.6 0.7 0.7 0.9 1.4 % Monomers 99.2 99.0 99.1 98.8 98.9 98.5 97.9 % LMW 0.4 0.5 0.3 0.5 0.5 0.6 0.7 Charge Variant Analysis by CEX-UPLC % Acid 18.9 19.3 21.8 27.0 19.9 22.5 27.2 % Principal 53.9 53.4 52.1 48.3 52.3 50.1 46.8 % Basic 27.3 27.4 26.1 24.8 27.8 27.4 26.0 Analysis of charge variants by iCIEF % Acid 31.9 NR 38.1 NR 43.7 NR NR 45.4 % Principal 54.7 NR 48.6 NR NR 44.3 NR NR 42.1 % Basic 13.5 NR 13.3 NR 12.0 NR NR 12.5 % relative potency by bioassay 126 NR NR 120 NR NR 99 CEX, cation exchange; DS, medicinal substance; HMW, high molecular weight; iCIEF, capillary focusing isoelectric imaging; LMW, low molecular weight; IMF, microflow; monomer, intact antibody; NR, not required; OD, optical density; PR, reversed phase; SE, size exclusion; UPLC, high-performance liquid chromatography Petition 870220073975, dated 08 / 17 / 2022, page 98 / 245 87 / 130 Table 14: Stability test of the formulation at 175 mg / mL stored under accelerated and stress conditions. Formulation mAbl at 175 mg / mL, L-histidine at 10 mM, sucrose at 5% (w / v), L-proline at 1.5% (w / v), polysorbate 80 at 0.2% (w / v), pH 6.0 Filling volume 1.2 mL Container / Seal 3 mL Type 1 glass vials with a 13 mm FluroTec® coated chlorobutyl 4432 / 50 stopper Storage at 25°C / 60% RH (months) Storage at 40°C (days) Assay 0 1 3 6 7 14 28 Color and appearance Approved Approved Approved Approved Approved Approved Approved Turbidity (DO increase at 405 nm) 0.00 0.00 0.00 0.00 0.00 0.00 0.01 Turbidity (NTU by Nephelometry) 6.72 6.77 6.82 6.99 6.90 7.30 7.56 pH 6.0 6.0 6.0 6.1 6.0 5.9 6.0 Subvisible Particulate Analysis by HIAC (# / mL) > 10 pm 10 NR 28 58 NR NR 97 > 25 pm 00 NR 13 16 NR NR 44 Subvisible Particulate Analysis by MFI (# / mL) 2-10 pm 144 NR 623 277 NR NR 1131 > 10 pm 22 NR 25 8 NR NR 531 > 25 pm 1 NR 3 2 NR NR 3 % Protein recovered by RP-UPLC 100 95 97 99 98 99 95 Purity by MCE-SDS Not reduced;% peak main 97.9 NR NR 97.4 NR NR 97.5 Reduced; % chain 100 NR NR 99.7 NR NR 99.4; 88 / 130 Petition 870220073975, dated 08 / 17 / 2022, page 99 / 245 Heavy + Light Purity by SE-UPLC % HMW 0.6 0.9 1.2 1.5 1.7 2.6 3.9 % Monomers 99.1 98.6 98.5 98.0 97.7 96.8 95.5 % LMW 0.4 0.5 0.3 0.6 0.6 0.7 0.6 Charge Variant Analysis by CEX-UPLC % Acid 18.7 18.9 21.3 26.2 19.7 22.0 23.9 % Principal 53.6 54.4 52.3 48.3 51.7 50.1 49.4 % Basic 27.8 26.7 26.4 25.5 28.6 27.9 26.7 Analysis of charge variants by iCIEF % Acid 31.8 NR 37.0 NR 45.0 NR NR 47.3 % Principal 54.6 NR 50.3 NR NR 43.3 NR NR 39.2 % Basic 13.6 NR 12.7 NR NR 11.7 NR NR 13.5 % relative potency by bioassay 102 NR NR 123 NR NR 86 CEX, cation exchange; DS, medicinal substance; HMW, high molecular weight; iCIEF, 89 / 130 capillary focusing isoelectric imaging, LMW, low molecular weight; IMF, microflow; Monomer, intact antibody; NR, not required; OD, optical density; PR, reversed phase; SE, size exclusion; UPLC, high-performance liquid chromatography Petition 870220073975, dated 08 / 17 / 2022, pp. 100 / 245 90 / 130

[0173] Example 12: Stability of mAbl formulations comprising histidine buffer, sucrose and polysorbate

[0174] Tables 15-24 summarize the storage stability of exemplary mAb1 formulations comprising 10 mM histidine buffer at pH 6.0, sucrose, and polysorbate. Table 15: Stability study of the mAb1 formulation stored at -80°C Formulation mAbl at 72.2 mg / mL, histidine at 10 mM, pH 6.0, sucrose at 5% (w / v) Filling volume 1.0 mL Container / Seal 5 mL polycarbonate bottle with silicone-coated polypropylene screw cap Shelf life at -80°C (months) Assay 0 1 3 6 9 12 Color and appearance Approved Approved Approved Approved Approved Approved Turbidity (DO increase at 405 nm) 0.00 0.00 0.00 0.00 0.00 0.00 pH 6.2 6.3 6.2 6.1 6.2 6.1 % protein recovered by RP-UPLC 100 94 95 98 95 94 Purity by MCE-SDS Not reduced; % Main Peak 99.5 NR NR 99.5 NR 99.2 Reduced;% Heavy + Light Chain 100 NR NR 100 NR 99.8 Purity by SE-UPLC % HMW 0.7 0.7 0.6 0.6 0.7 0.6 % Native 98.7 98.8 98.9 98.7 98.9 98.8 % LMW 0.6 0.6 0.5 0.6 0.4 0.6 Charge Variant Analysis by CEX-UPLC % Acid 22.2 22.2 22.6 23.2 24.1 23.5 % Principal 49.7 49.8 48.9 46.8 45.2 44.1 % Basic 28.1 28.0 28.5 30.0 30.7 32.4 Charge Variant Analysis Charge by iCIEF % Acid 38.9 NR NR 39.1 NR 37.5 % Principal 56.5 NR NR 56.2 NR 57.2 % Basic 4.6 NR NR 4.7 NR 5.3 % Relative potency (bioassay) 95 NR NR 81 NR 120; CEX = cation exchange; HMW = high molecular weight; iCIEF = Capillary focusing isoelectric imaging; LMW = low molecular weight; MCE-SDS = Microchip dodecyl capillary electrophoresis Petition 870220073975, dated 08 / 17 / 2022, page 101 / 245 91 / 130 sodium sulfate; IMF = microflow imaging; NR = Not required; OD = optical density; RH = relative humidity; PR = phase reversal; SE = size exclusion; UPLC = high-performance liquid chromatography Table 16: Stability study of the mAb1 formulation stored at -30°C Formulation mAb1 at 72.2 mg / mL, histidine at 10 mM, pH 6.0, sucrose at 5% (w / v) Filling volume 1.0 mL Container / Seal 5 mL polycarbonate bottle with silicone-coated polypropylene screw cap Storage duration at -30°C (months) Assay 0 1 3 6 9 12 Color and appearance Approved Approved Approved Approved Approved Approved Turbidity (DO increase at 405 nm) 0.00 0.00 0.00 0.00 0.00 0.01 pH 6.2 6.3 6.2 6.1 6.2 6.1 % protein recovered by RP-UPLC 100 93 95 100 96 96 Purity by MCE-SDS Not reduced; % Main Peak 99.5 NR NR 99.1 NR 99.2 Reduced;% Heavy + Light Chain 100 NR NR 100 NR 99.8 Purity by SE-UPLC % HMW 0.7 0.7 0.6 0.7 0.7 0.6 % Native 98.7 98.8 99.0 98.7 98.9 98.8 % LMW 0.6 0.5 0.4 0.6 0.4 0.6 Charge Variant Analysis by CEXUPLC % Acid 22.2 22.5 22.5 23.4 24.2 23.4 % Principal 49.7 49.6 48.9 46.6 45.6 44.1 % Basic 28.1 28.0 28.6 30.0 30.2 32.5 Charge Variant Analysis by iCIEF % Acid 38.9 NR 38.2 NR 37.8 % Main 56.5 NR NR 56.3 NR 56.6 % Basic 4.6 NR NR 5.5 NR 5.7; Table 17: Stability study of the mAb1 formulation stored at -20°C Petition 870220073975, dated 08 / 17 / 2022, page 102 / 245 92 / 130 Formulation mAbl at 72.2 mg / mL, histidine at 10 mM, pH 6.0, sucrose at 5% (w / v) Filling volume 1.0 mL Container / Seal 5 mL polycarbonate bottle with silicone-coated polypropylene screw cap Storage duration at -20°C (months) Assay 0 1 3 6 9 12 Color and appearance Approved Approved Approved Approved Approved Approved Turbidity (DO increase at 405 nm) 0.00 0.00 0.01 0.01 0.00 0.01 pH 6.2 6.3 6.2 6.1 6.1 6.2 % protein recovered by RP-UPLC 100 95 97 101 98 98 Purity by MCE-SDS Not reduced; % Main Peak 99.5 NR NR 99.5 NR 99.3 Reduced;% Heavy + Light Chain 100 NR NR 100 NR 99.9 Purity by SE-UPLC % HMW 0.7 0.7 0.7 0.7 0.8 0.7 % Native 98.7 98.8 98.9 98.6 98.8 98.8 % LMW 0.6 0.5 0.5 0.7 0.4 0.6 Charge Variant Analysis by CEX-UPLC % Acid 22.2 22.4 22.3 22.4 24.6 23.4 % Principal 49.7 49.7 49.2 47.6 45.5 44.2 % Basic 28.1 27.9 28.5 30.0 30.0 32.5 Charge Variant Analysis Charge by iCIEF % Acid 38.9 NR NR 38.6 NR 38.6 % Principal 56.5 NR NR 56.8 NR 56.2 % Basic 4.6 NR NR 4.6 NR 5.3 % Relative potency (bioassay) 95 NR NR 96 NR 111; Petition 870220073975, dated 08 / 17 / 2022, page 103 / 245 Table 18: Stability study of the mAbl formulation - Effect under accelerated conditions Formulation mAbl at 72.2 mg / mL, histidine at 10 mM, pH 6.0, sucrose at 5% (w / v) Filling volume 1.0 mL Container / Seal 5 mL polycarbonate bottle with silicone-coated polypropylene screw cap Storage at 5°C (days) Storage at 25°C / 60% RH (days) Storage at 40°C / 75% RH (days) Assay T = 0 28 56 14 28 14 28 Color and appearance Approved Approved Approved Approved Approved Approved Approved Approved Turbidity (DO increase at 405 nm) 0.00 0.01 0.01 0.02 0.01 0.03 0.05 pH 6.2 6.2 6.1 6.2 6.2 6.2 6.1 % of protein recovered by RP-UPLC 100 96 100 97 100 105 113 Purity by MCE-SDS Not reduced; % Main peak 99.5 NR 99.1 NR 99.5 NR 99.1 Reduced; % Heavy + light chain 100 NR 100 NR 99.2 NR 98.7 Purity by SE-UPLC % HMW 0.7 0.9 1.1 1.4 1.7 3.1, 4.6% Native 98.7 98.5 98.4 98.0 97.7 96.1 94.6% LMW 0.6 0.6 0.6 0.7 0.7 0.8 0.9 Charge variant analysis by CEX-UPLC % Acid 22.2 22.2 22.3 22.0 22.6 25.6 30.8% Principal 49.7 49.8 48.9 49.6 49.1 46.0 41.7% Basic 28.1 28.1 28.8 28.4 28.3 28.3 27.5 Charge variant analysis by iCIEF % Acid 38.9 NR 39.0 NR 41.1 NR 58.2% Main 56.5 NR 56.8 NR 53.8 NR 36.3 % Basic 4.6 NR 4.2 NR 5.1 NR 5.5 % relative power (bioassay) 95 NR 95 NR 84 NR 87. 93 / 130 Petition 870220073975, dated 08 / 17 / 2022, page 104 / 245 Table 19: Stability study of the mAbl formulation stored at -80°C Formulation mAbl at 25 mg / mL, histidine at 10 mM, pH 6.0, sucrose at 10% (w / v), polysorbate 80 at 0.1% Filling volume 1.0 mL Container / Seal 5 mL polycarbonate bottle with silicone-coated polypropylene screw cap Storage duration at -80°C (months) Assay 0 1 3 6 9 12 Color and appearance Approved Approved Approved Approved Approved Approved Turbidity (DO increase at 405 nm) 0.00 0.00 0.00 0.00 0.01 0.01 pH 6.1 6.1 6.1 6.0 6.0 6.0 % protein recovered by RP-UPLC 100 100 97 95 100 99 Purity by MCE-SDS Not reduced; % Main peak 99.5 NR NR 99.4 NR 99.5 Reduced;% Heavy + Light Chain 100 NR NR 100 NR 100 Purity by SE-UPLC % HMW 0.8 0.8 0.8 0.8 0.8 0.8 % Native 98.6 98.7 98.6 98.6 98.8 98.7 % LMW 0.6 0.5 0.6 0.6 0.5 0.5 Charge Variant Analysis by CEX-UPLC % Acid 22.8 23.0 23.3 23.5 23.9 25.4 % Principal 47.3 47.3 46.2 45.3 44.2 44.2 % Basic 30.0 29.8 30.6 31.2 31.9 30.4 Charge Variant Analysis by iCIEF % Acid 40.1 NR NR 38.1 NR 41.3 % Principal 56.1 NR NR 57.4 NR 54.1 % Basic 3.8 NR NR 4.6 NR 4.6 % relative potency (bioassay) 112 NR NR 130 NR 107; 94 / 130 Petition 870220073975, dated 08 / 17 / 2022, page 105 / 245 Table 20: Stability study of the mAbl formulation stored at -30°C Formulation mAbl at 25 mg / mL, histidine at 10 mM, pH 6.0, sucrose at 10% (w / v), polysorbate 80 at 0.1% Filling volume 1.0 mL Container / Seal 5 mL polycarbonate bottle with silicone-coated polypropylene screw cap Storage duration at -30°C (months) Assay 0 1 3 6 9 12 Color and appearance Approved Approved Approved Approved Approved Approved Turbidity (DO increase at 405 nm) 0.00 0.00 0.00 0.00 0.01 0.01 pH 6.1 6.1 6.1 6.0 6.1 6.0 % protein recovered by RP-UPLC 100 100 102 97 100 102 Purity by MCE-SDS Not reduced; % Main peak 99.5 NR NR 99.1 NR 99.5 Reduced;% Heavy + Light Chain 100 NR NR 100 NR 100 Purity by SE-UPLC % HMW 0.8 0.8 0.8 0.8 0.8 0.8 % Native 98.6 98.7 98.6 98.7 98.8 98.7 % LMW 0.6 0.5 0.6 0.6 0.4 0.6 Charge Variant Analysis by CEX-UPLC % Acid 22.8 23.1 22.9 23.5 23.7 25.1 % Principal 47.3 47.1 46.5 45.3 44.4 44.4 % Basic 30.0 29.8 30.6 31.2 31.9 30.5 Charge Variant Analysis by iCIEF % Acid 40.1 NR 38.0 NR 41.3 % Main 56.1 NR NR 57.3 NR 53.3 % Basic 3.8 NR NR 4.7 NR 5.4; 95 / 130 Petition 870220073975, dated 08 / 17 / 2022, page 106 / 245 Table 21: Stability study of the mAbl formulation stored at -20°C Formulation mAbl at 25 mg / mL, histidine at 10 mM, pH 6.0, sucrose at 10% (w / v), polysorbate 80 at 0.1% Filling volume 1.0 mL Container / Seal 5 mL polycarbonate bottle with silicone-coated polypropylene screw cap Storage duration at -20°C (months) Assay 0 1 3 6 9 12 Color and appearance Approved Approved Approved Approved Approved Approved Turbidity (DO increase at 405 nm) 0.00 0.00 0.00 0.00 0.01 0.00 pH 6.1 6.1 6.1 6.0 6.1 6.0 % protein recovered by RP-UPLC 100 99 102 97 106 102 Purity by MCE-SDS Not reduced; % Main peak 99.5 NR NR 99.4 NR 99.4 Reduced;% Heavy + Light Chain 100 NR NR 100 NR 99.0 Purity by SE-UPLC % HMW 0.8 0.8 0.8 0.8 0.7 0.8 % Native 98.6 98.7 98.6 98.7 98.8 98.7 % LMW 0.6 0.5 0.6 0.6 0.4 0.6 Charge Variant Analysis by CEX-UPLC % Acid 22.8 22.9 23.5 23.6 24.3 25.2 % Principal 47.3 47.3 46.0 45.2 43.8 43.9 % Basic 30.0 29.8 30.6 31.2 31.9 30.8 Charge Variant Analysis Charge by iCIEF % Acid 40.1 NR NR 38.4 NR 41.6 % Principal 56.1 NR NR 57.9 NR 53.3 % Basic 3.8 NR NR 3.7 NR 5.1 % Relative potency (bioassay) 112 NR NR 120 NR 131; 96 / 130 Petition 870220073975, dated 08 / 17 / 2022, page 107 / 245 Table 22: Stability study of the mAbl formulation - Effect under accelerated conditions Formulation mAbl at 25 mg / mL, histidine at 10 mM, pH 6.0, sucrose at 10% (w / v), polysorbate 80 at 0.1% Filling volume 1.0 mL Container / Seal 5 mL polycarbonate bottle with silicone-coated polypropylene screw cap No storage Storage at 5°C (days) Storage at 25°C / 60% RH (days) Storage at 40°C / 75% RH (days) Assay T = 0 28 56 14 28 14 28 Color and appearance Approved Approved Approved Approved Approved Approved Approved Turbidity (DO increase at 405 nm) 0.00 0.00 0.00 0.00 0.00 0.01 0.03 pH 6.1 6.1 6.0 6.1 6.1 6.0 6.0 % of protein recovered by RP-UPLC 100 101 102 106 107 113 114 Purity by MCESDS Not reduced; % Main peak 99.5 NR 99.1 NR 99.5 NR 99.3 Reduced;% Heavy + Light Chain 100 NR 100 NR 100 NR 99.1 Purity by SE-UPLC % HMW 0.8 0.7 0.7 0.9 0.9 3.4 5.0 % Native 98.6 98.8 98.7 98.6 98.6 95.7 93.7 % LMW 0.6 0.5 0.6 0.5 0.6 0.9 1.3 Charge Variant Analysis by CEXUPLC % Acid 22.8 22.8 23.3 22.6 22.8 29.6 32.5 % Principal 47.3 47.3 46.1 47.2 47.0 39.5 36.4 % Basic 30.0 29.9 30.6 30.2 30.2 30.9 31.1 Analysis of charge variants by iCIEF % Acid 40.1 NR 39.8 NR 40.2 NR 53.7 % Principal 56.1 NR 57.0 NR 55.9 NR 42.2 % Basic 3.8 NR 3.2 NR 3.9 NR 4.0 % relative potency (bioassay) 112 NR 103 NR 91 NR 79; 97 / 130 Petition 870220073975, dated 08 / 17 / 2022, page 108 / 245 Table 23: Stability study of the mAbl formulation stored at 5 °C Formulation mAbl at 25 mg / mL, histidine at 10 mM, sucrose at 10% (w / v), polysorbate 80 at 0.1% (w / v), pH 6.0 Filling volume 1.0 mL Container / Seal 2 mL borosilicate glass bottles type 1 with 13 mm West S2-451 4432 / 50 GRY B2-40 FluroTec® coated stoppers Storage duration at 5°C (months) Assay 0 1 3 6 9 12 Color and appearance Approved Approved Approved Approved Approved Approved Turbidity (DO increase at 405 nm) 0.00 0.00 0.00 0.01 0.01 0.00 pH 6.2 6.1 6.1 6.1 6.0 6.1 Particulate analysis by MFI (particles / mL) 2 to 10 pm 823 NR NR 2169 NR 29331 > 10 pm 15 NR NR 10 NR 56 > 25 pm 00 NR NR 3 NR 5 % protein recovered by RP-UPLC 100 100 98 101 102 90 Purity by MCE-SDS Not reduced; % main peak 99.4 NR NR 99.5 NR 99.2 Reduced;% Heavy + Light Chain 100 NR NR 100 NR 100 Purity by SE-UPLC % HMW 0.7 0.7 0.7 0.8 0.9 0.9 % Native 98.7 98.8 98.7 98.6 98.6 98.5 % LMW 0.6 0.5 0.6 0.6 0.5 0.6 Charge Variant Analysis by CEX-UPLC % Acid 23.0 22.8 23.5 22.4 24.1 22.4 % Principal 48.5 48.6 46.2 47.2 44.4 45.5 % Basic 28.6 28.6 30.4 30.4 31.5 32.1 Charge Variant Analysis by iCIEF % Acid 39.8 NR NR 39.2 NR 40.1 % Principal 56.7 NR NR 56.5 NR 55.7 % Basic 3.4 NR NR 4.3 NR 4.1 % Relative potency (bioassay) 111 NR NR 132 NR 124; 98 / 130 Petition 870220073975, dated 08 / 17 / 2022, page 109 / 245 Table 24: Stability study of mAbl formulation stored under accelerated conditions Formulation mAbl at 25 mg / mL, histidine at 10 mM, sucrose at 10% (w / v), polysorbate 80 at 0.1% (w / v), pH 6.0 Filling volume 1.0 mL Container / Seal 2 mL borosilicate glass bottles type 1 with 13 mm West S2-451 4432 / 50 GRY B2-40 stoppers coated with FluroTec® Storage at 25°C / 60% RH (months) Storage at 45°C (days) Assay 0 0.5 1 3 7 14 28 Color and appearance Approved Approved Approved Approved Approved Approved Approved Turbidity (DO increase at 405 nm) 0.00 0.01 0.01 0.00 0.01 0.00 0.02 pH 6.2 6.1 6.1 6.1 6.2 6.1 6.1 Particulate analysis by MFI (particles / mL) 2 to 10 pm 823 NR NR 1056 NR NR 521 > 10 pm 15 NR NR 23 NR NR 83 > 25 pm 0 0 NR NR 3 NR NR 4 % protein recovered by RP-UPLC 100 99 100 99 99 100 99 Purity by MCESDS Not reduced; % main peak 99.4 NR NR 99.4 NR NR 99.2 Reduced;% Heavy + Light Chain 100 NR NR 100 NR NR 98.6 Purity by SE-UPLC % HMW 0.7 0.8 0.9 1.1 1.6 3.2 8.5 % Native 98.7 98.6 98.5 98.1 97.6 95.9 89.7 % LMW 0.6 0.6 0.6 0.8 0.8 1.0 1.8 Charge Variant Analysis by CEX-UPLC % Acid 23.0 22.6 23.1 25.8 24.7 27.3 34.0 % Principal 48.5 48.7 48.1 44.5 46.2 44.1 35.9 % Basic 28.6 28.8 28.9 29.8 29.1 28.6 30.2 Analysis of charge variants by CIEF % Acid 39.8 NR NR 47.1 NR NR 66.9 % Principal 56.7 NR NR 49.5 NR NR 30.2 % Basic 3.4 NR NR 3.5 NR NR 3.0 % relative potency by bioassay 111 NR NR 94 NR NR 63; 99 / 130

[0175] Tables 25-27 summarize the stress stability of exemplary formulations. Table 25: Stability research of the mAb1 formulation - Effect of stress conditions Petition 870220073975, dated 08 / 17 / 2022, page 110 / 245 Formulation mAbl at 72.2 mg / mL, histidine at 10 mM, pH 6.0, sucrose at 5% (w / v) Filling volume 1.0 mL Container / Seal 5 mL polycarbonate bottle with silicone-coated polypropylene screw cap Agitation (min) Freeze / Thaw (cycles) Assay T = 0 10 15 4 8 Color and appearance Approved Approved Approved Approved Approved Turbidity (DO increase at 405 nm) 0.00 0.00 0.09 0.01 0.00 pH 6.2 6.2 6.2 6.3 6.3 % protein recovered by RP-UPLC 100 100 99 95 95 Purity by MCESDS Not reduced; % Main Peak 99.5 NR 99.2 NR 99.4 Reduced;% Heavy + Light Chain 100 NR 100 NR 100 Purity by SE-UPLC % HMW 0.7 0.8 4.6 0.6 0.7 % Native 98.7 98.7 94.9 98.9 98.8 % LMW 0.6 0.5 0.5 0.5 0.5 Charge Variant Analysis by CEX-UPLC % Acid 22.2 22.2 21.9 22.0 22.2 % Principal 49.7 49.6 49.9 49.7 49.6 % Basic 28.1 28.2 28.2 28.3 28.2 Charge Variant Analysis by CIEF % Acid 38.9 NR 40.8 NR 39.0 % Main 56.5 NR 54.7 NR 56.5 % Basic 4.6 NR 4.6 NR 4.5 % relative potency (bioassay) 95 NR 87 NR 89; 100 / 130 Table 26: Stability study of mAbl formulation - Effect of stress conditions Petition 870220073975, dated 08 / 17 / 2022, page 111 / 245 Formulation mAbl at 25 mg / mL, histidine at 10 mM, pH 6.0, sucrose at 10% (w / v), polysorbate 80 at 0.1% (w / v) Filling volume 1.0 mL Container / Seal 5 mL polycarbonate bottle with silicone-coated polypropylene screw cap Stress-free Shaking (minutes) Freezing / Thawing (cycles) Assay T = 0 60 120 4 8 Color and appearance Approved Approved Approved Approved Approved Turbidity (DO increase at 405 nm) 0.00 0.00 0.00 0.00 0.00 pH 6.1 6.1 6.1 6.1 6.1 % protein recovered by RP-UPLC 100 101 100 99 100 Purity by MCE-SDS Not reduced; % Main peak 99.5 NR 99.5 NR 99.6 Reduced;% Heavy + Light Chain 100 NR 100 NR 100 Purity by SE-UPLC % HMW 0.8 0.8 0.7 0.8 0.8 % Native 98.6 98.7 98.7 98.7 98.6 % LMW 0.6 0.6 0.6 0.5 0.6 Charge Variant Analysis by CEX-UPLC % Acid 22.8 22.8 22.8 22.9 22.7 % Principal 47.3 47.2 47.3 47.1 47.2 % Basic 30.0 30.0 29.2 30.1 30.1 Charge Variant Analysis by iCIEF % Acid 40.1 NR 39.6 NR 40.0 % Principal 56.1 NR 56.9 NR 56.3 % Basic 3.8 NR 3.5 NR 3.6 % relative potency (bioassay) 112 NR 106 NR 137; 101 / 130 Petition 870220073975, dated 08 / 17 / 2022, page 112 / 245 Table 27: Stability study of mAbl formulation - Effect of stress conditions Formulation mAbl at 25 mg / mL, histidine at 10 mM, sucrose at 10% (w / v), polysorbate 80 at 0.1% (w / v), pH 6.0 Filling volume 1.0 mL Container / Seal 2 mL borosilicate glass bottles type 1 with 13 mm West S2-451 4432 / 50 GRY B2-40 FluroTec® coated stoppers Agitation (min) Freeze / Thaw (cycles) Assay 0 60 120 4 8 Color and appearance Approved Approved Approved Approved Approved Turbidity (DO increase at 405 nm) 0.00 0.00 0.00 0.00 0.00 pH 6.2 6.2 6.2 6.2 6.2 Particulate analysis by MFI (particles / mL) 2 to 10 pm 823 NR 1170 NR 1404 > 10 pm 15 NR 38, NR 67 > 25 pm 00 NR 00 NR 2 % protein recovered by RP-UPLC 100 100 101 101 101 Purity by MCE-SDS Not reduced; % main peak 99.4 NR 99.4 NR 99.3 Reduced;% Heavy + Light Chain 100 NR 100 NR 100 Purity by SE-UPLC % HMW 0.7 0.6 0.7 0.7 0.7 % Native 98.7 98.8 98.7 98.7 98.6 % LMW 0.6 0.6 0.6 0.6 0.7 Charge Variant Analysis by CEX-UPLC % Acid 23.0 22.9 22.8 22.8 22.7 % Principal 48.5 48.3 48.4 48.4 48.5 % Basic 28.6 28.8 28.8 28.8 28.8 Charge Variant Analysis by iCIEF % Acid 39.8 NR 39.6 NR 39.6 % Principal 56.7 NR 56.9 NR 56.9 % Basic 3.4 NR 3.4 NR 3.6 % relative potency per bioassay 111 NR 90 NR 129; 102 / 130 Petition 870220073975, dated 08 / 17 / 2022, page 113 / 245 Table 28: Accelerated and stress stability of mAbl formulations at 50 mg / mL and 25 mg / mL Formulation mAbl at 50 mg / mL, histidine at 10 mM, sucrose at 10% (w / v), polysorbate 80 at 0.1% (w / v), pH 6.0 mAbl at 25 mg / mL, histidine at 10 mM, sucrose at 10% (w / v), polysorbate 80 at 0.1% (w / v), pH 6.0 Filling volume 1.0 mL 1.0 mL Container / Seal 2 mL borosilicate glass bottles type 1 with 13 mm West S2-451 4432 / 50 GRY B2-40 FluroTec® coated stoppers 2 mL borosilicate glass bottles type 1 with 13 mm West S2-451 4432 / 50 GRY B2-40 coated stoppers FluroTec® Assay T = 0 25°C / 60% RH 1 month 45°C 28 days T = 0 25°C / 60% RH 1 month 45°C 28 days Color and appearance Approved Approved Approved Approved Approved Approved Turbidity (DO increase at 405 nm) 0.00 0.01 0.02 0.00 0.01 0.02 pH 6.0 6.0 6.1 6.2 6.1 6.1 % protein recovered by RP-UPLC 100 103 113 100 100 99 Purity by SE-UPLC % HMW 1.9 4.0 10.8 0.7 0.9 8.5 % monomers 97.5 95.2 88.0 98.7 98.5 89.7 % LMW 0.6 0.8 1.2 0.6 0.6 1,8. Analysis of charge variants by CEXUPLC % Acid 24.6 29.0 37.1 23.0 23.1 34.0 % Principal 49.9 42.5 33.2 48.5 48.1 35.9 % Basic 25.6 28.5 29.7 28.6 28.9 30.2 103 / 130 Petition 870220073975, dated 08 / 17 / 2022, pp. 114 / 245 104 / 130 Example 13: Proven acceptable range (PAR) study

[0176] During the manufacture of the mAb1 drug (DP), variations in the composition of the DP may occur. These variations may include the concentration of the active ingredient, the concentration of excipients, and / or the pH of the formulation. Since changes in any of these parameters can potentially affect the stability or potency of the drug, proven acceptable range (PAR) studies were conducted to evaluate whether variations in the composition of the DP, within defined limits, would affect the stability or potency of the mAb1 DP.

[0177] Two design-of-experiment (DOE) studies were used to evaluate the effect of each formulation parameter, as well as the interactions, on formulation stability: • A pre-PAR fractional factorial design study with accelerated and stress stability assessment to identify critical formulation parameters that may affect the stability of the mAb1 DP; • A comprehensive PAR study using factorial design, including critical formulation parameters identified in the PréPAR study, with long-term stability validity to demonstrate acceptable ranges for the formulation parameters. Conception of the pre-PAR Study

[0178] To evaluate critical and / or interaction parameters of formulations in DP composition that may be important for product quality, a fractional factorial DOE was applied to examine the accelerated and stress stability of the formulations by varying all formulation parameters, including protein concentration (± 10%), buffer and stabilizer concentrations (± 20%), surfactant concentration (± 50%), and pH (± 0.3 units). The ranges of formulation parameters tested were defined as equal to or greater than the specification acceptance criteria and Petition 870220073975, dated 08 / 17 / 2022, pp. 115 / 245 105 / 130 manufacturing experience. The study was designed with statistical software using a fractional factorial IV experiment with 2Λ(6-2) resolutions. Along with four target formulations as central points, the study included 20 runs, as shown in Table 29. Table 29: Formulations tested in the pre-PAR study Formulation [mAbl] mg / mL [histidine] mM % sucrose (w / v) % L-proline (w / v) % polysorbate 80 (w / v) pH 1 45 12 4 1.2 0.3 6.3 2 45 12 4 1.8 0.3 5.7 3 45 8 6 1.8 0.3 6.3 4 50 10 5 1.5 0.2 6.0 5 45 8 4 1.8 0.1 6.3 6 55 8 6 1.2 0.1 6.3 7 55 8 4 1.2 0.3 6.3 8 50 10 5 1.5 0.2 6.0 9 55 12 6 1.8 0.3 6.3 10 45 12 6 1.2 0.1 6.3 11 55 8 4 1.8 0.3 5.7 12 55 8 6 1.8 0.1 5.7 13 45 12 6 1.8 0.1 5.7 14 55 12 6 1.2 0.3 5.7 15 50 10 5 1.5 0.2 6.0 16 45 8 6 1.2 0.3 5.7 17 55 12 4 1.8 0.1 6.3 18 45 8 4 1.2 0.1 5.7 19 50 10 5 1.5 0.2 6.0 20 55 12 4 1.2 0.1 5.7

[0179] All 20 formulations under accelerated conditions and stress conditions (25°C, 37°C, freeze / thaw [F / T] and agitation) were characterized and evaluated for physicochemical properties and stability, including visual inspection, pH, turbidity, osmolarity, conductivity, purity, protein concentration and recovery, charge variant analysis and subvisible particulate analysis. Results of the Pre-PAR Study Petition 870220073975, dated 08 / 17 / 2022, pp. 116 / 245 106 / 130

[0180] All 20 formulations showed no alteration after agitation or F / T stress. The results of incubation at 25°C and 37°C were analyzed by a regression model (JMP fit model with standard quadratic personality and emphasis on leverage effect). Statistical analysis of the main variables and interaction of all experimental formulations in relation to critical quality attributes revealed that pH, protein concentration, and sucrose concentration were important for product quality. The two product quality attributes that had an impact were HMW species and acid load variants. It was shown that other formulation parameters, including histidine, proline, or polysorbate 80 concentrations, within the tested ranges, did not have a statistically significant impact on product quality.Under accelerated conditions, there were no secondary or major interactions that impacted the stability of the formulation. The results of the pre-PAR study indicated that pH, mAb1 concentration, and sucrose concentration were critical for the stability of the mAb1 formulation and were considered the critical formulation parameters for the 50 mg / mL mAb1 formulation.

[0181] Although pH, ​​mAb1 concentration, and sucrose concentration were identified as critical formulation parameters, the impact of these three factors on quality attributes was minimal. Based on statistical analysis, variation in the pH range of 5.7-6.3, 45-55 mg / mL mAb1, and / or 4-6% sucrose likely had a <15% impact on the formation of HMW species and acid load variants.

[0182] To confirm the impact on long-term storage stability under the recommended storage conditions for DP, the three critical formulation parameters a Petition 870220073975, dated 08 / 17 / 2022, page 117 / 245 107 / 130 following: pH, mAbl concentration and sucrose concentration, were evaluated in a PAR study with long-term storage stability. PAR Study Design

[0183] A full factorial design DOE was applied to examine the long-term shelf-life stability of formulations with varying pH (± 0.3 units), protein concentration (± 10%), and sucrose concentration (± 20%), resulting in eight experimental runs (Table 30); a reference formulation (formulation 3, the target formulation in Table 30) was included as the central point formulation. Table 30: Formulations tested in PAR studies Formulation [mAbl] mg / mL [histidine] mM % sucrose (w / v) % L-proline (w / v) % polysorbate 80 (w / v) pH 1 55 10 6 1.5 0.2 5.7 2 45 10 6 1.5 0.2 5.7 3 50 10 5 1.5 0.2 6.0 4 55 10 6 1.5 0.2 6.3 5 55 10 4 1.5 0.2 6.3 6 55 10 4 1.5 0.2 5.7 7 45 10 4 1.5 0.2 6.3 8 45 10 4 1.5 0.2 5.7 9 45 10 6 1.5 0.2 6.3

[0184] The full factorial design study allows estimation of all terms of the main effect, as well as the interaction terms. The ranges of formulation parameters tested, defined as equal to or greater than the specification acceptance criteria and manufacturing experience, remained the same as in the pre-PAR study.

[0185] The stability of the experimental formulations was compared with the stability of a reference formulation at pH 6.0 containing all formulation components at their nominal concentrations (F3). The PAR studies used a batch of mAb1 DS manufactured using a representative commercial manufacturing process. The formulations Petition 870220073975, dated 08 / 17 / 2022, pp. 118 / 245 108 / 130 of DP were filled into 10 mL Schott type 1 borosilicate glass bottles, fitted with 20 mm West S2-451 4432 / 50 GRY B2-40 stoppers coated with FluroTec® (DP commercial representation) and evaluated for long-term storage stability at 2-8 °C. The formulations were studied according to the analysis plan in Table 31. Table 31: Analysis plan for the PAR study Assay | Samples Analyzed | Quality Attributes | Appearance | All | Color, visible particulates | Turbidity (DO increase at 405 nm) | All | Color, particulates, clarity | Turbidity by Nephelometry | t = 0, 6, 12, 18, 24 and 36 months at 5°C | Clarity (Solution precipitation, Particulates, Opalescence) | pH | All | pH | Total protein content by RP-UPLC | t = 0 | Protein concentration | Osmolality | t = 0 | Solute concentration | Conductivity | t = 0 | Conducting property | Purity by SE-UPLC | All | Molecular weight variants: % HMW, % monomer, % LWM | Charge variant analysis by CIEF | t = 0, 6, 12, 18, 24 and 36 months at 5°C | Charge isoforms: % acidic, % principal, % basic | Particulate material (Light Obscuring) t = 0, 6, 12, 18, 24 and 36 months at 5°C Subvisible particulates.Acceptance criteria established at USP <788> (< 6000 particles / container for particles > 10 µm and < 600 particles / container for particles > 25 μm) Particulate matter (Microflow Imaging, MFI) t = 0, 6, 12, 18, 24 and 36 months at 5°C Subvisible particulates (For information only) Bioassay t = 0, 6, 12, 18, 24 and 36 months at 5°C Potency Acceptance criteria: 50-150% of the reference standard. Results of the PAR Study Effect on the Formation of HMW Species Petition 870220073975, dated 08 / 17 / 2022, p. 119 / 245 109 / 130

[0186] There was no significant increase in % HMW, as measured by SE-UPLC up to 12 months at 2-8 °C for all 9 formulations. All values ​​were well below the maximum specification and no significant increase in % HMW was observed over time.

[0187] It was observed that the formation of HMW species for the mAb1 DP at 50 mg / mL at 2-8 °C was extremely slow. For up to 12 months, the maximum variation in the relative amount of % HMW in the 9 formulations was ~0.2%. Since the variation in % HMW was minimal and the monomer concentration could be considered constant, the aggregation of monomers to HMW species could be simplified as a zero-magnitude reaction. Therefore, a simplified linear model was used to analyze the stability data of % HMW. By linearly fitting the % HMW over time, the rate of formation of HMW species was derived for each formulation.

[0188] The rate was analyzed in relation to the main factors, as well as all interaction terms, using a regression model (JMP fit model with standard least squares personality and emphasis on leverage effect). The resulting regression model was statistically significant, with R2 of 0.74. mAb1 concentration, pH, and time were statistically significant, however, the effect on the % formation of HMW species was statistically insignificant, contributing only 0.1%.

[0189] Therefore, these factors, pH 5.7 to 6.3, mAb1 concentration of 45 to 55 mg / mL and sucrose concentration of 4-6%, had no practical relevance for the stability of % HMW between 2-8°C.

[0190] The % of HMW in all 9 formulations at the time point of up to 12 months was well below the defined acceptance criteria. Petition 870220073975, dated 08 / 17 / 2022, pp. 120 / 245 110 / 130 of 4%, and therefore within the release and specifications at the end of the shelf life. Furthermore, the linear model predicted that after 24 months of storage, the shelf life at 2°C - 8°C, the % of HMW, which ranged from 0.6% to 0.8%, would also be well below the specification limit.

[0191] Based on long-term storage stability data, variations in critical formulation parameters within the studied ranges were found to have no significant impact on the stability of the mAb1 formulation. The 50 mg / mL mAb1 formulation was robust with respect to the formation of HMW species within the tested formulation composition range. Effect on the Formation of Acid Charge Variants

[0192] There was no significant increase in the % of acid load variants measured up to 12 months at 2-8 °C for all 9 formulations. All values ​​were below the maximum specification and no significant increase in the % of acid load variants was observed over time.

[0193] The mAb1 formulation was considered to be robust with regard to the formation of acid charge variants within the composition range of the formulation tested. Effect on General Quality Attributes

[0194] The effect of pH, mAb1 and sucrose concentration, as well as storage time, on other general DP quality attributes, including appearance, pH, turbidity, subvisible particulates, protein recovery, % monomer and % LMW by SEC, % principal and % base charge variants by iCIEF, and bioactivity, was studied. All values ​​were within specifications and no significant variation over time or difference between PAR formulations was observed: • No visible precipitate or particulate matter was detected by Petition 870220073975, dated 08 / 17 / 2022, pp. 121 / 245 111 / 130 means of visual inspection or turbidity measurements (DO at 405 nm and nephelometry); • No statistically significant changes were observed in protein recovery (RP-UPLC); • The pH of the formulations was stable; • No significant increases in subvisible particulate matter were observed, and no significant differences in subvisible particulate matter counts were observed between the PAR study formulations. For subvisible particulate matter measured by HIAC, all values ​​were below the acceptable limits established by the USP. <788> , and no significant variation in subvisible particulates between the formulations was observed. Furthermore, subvisible particles were also measured by IMF. No significant variation in subvisible particles was observed between the formulations. • The bioassay results were within the specification limit for all formulations during storage.

[0195] The results demonstrate that variations in critical formulation parameters (pH, mAb1 concentration, and sucrose concentration) within the studied ranges do not have a significant impact on the stability of the mAb1 formulation. The 50 mg / mL mAb1 formulation is robust with respect to overall quality attributes within the tested formulation composition range. Effect of Freezing and Thawing on the Stability of PAR Formulations

[0196] The physical and chemical stability of the mAb1 50 mg / mL formulation, examined following two freeze-thaw cycles, was not affected by the variation in critical formulation parameters, i.e., a change of ± 0.3 pH units in Petition 870220073975, dated 08 / 17 / 2022, pp. 122 / 245 112 / 130 relative to the reference mAb1, a variation of ± 10% in mAb1 concentration and / or a variation of ± 20% in sucrose.

[0197] The following effects were observed: • No precipitate was detected by visual inspection or turbidity measurements (DO at 405 nm); • No protein loss was observed (RP-UPLC); • The pH of the formulations remained constant; • No significant differences were observed in subvisible particulate counts, determined by light obscuration (HIAC) or microflow imaging (MFI), between the study formulations. There was a slight increase in subvisible particles after 2 F / T cycles, which can be removed by filtration through a 0.22 μm filter before filling the DP. • No appreciable changes in purity, as determined by SE-UPLC, were observed in any of the formulations after two freeze-thaw cycles; • No appreciable variation was observed in the distribution of load variants, as determined by iCIEF, in all formulations after two freeze-thaw cycles. • The bioassay results demonstrated that mAb1 activity was maintained in all formulations subjected to 2 freeze-thaw cycles. Conclusions

[0198] Pre-PAR and PAR studies based on Design of Experiments (DOE) were used to evaluate the effect of formulation parameters as well as interactions on formulation stability. The pre-PAR study under accelerated stability and stress identified pH, mAb1 concentration, and sucrose concentration as critical formulation parameters. A factorial PAR study Petition 870220073975, dated 08 / 17 / 2022, pp. 123 / 245 The 113 / 130 complete long-term stability study demonstrated that variation in critical formulation parameters, within the study ranges, does not affect the quality of the mAb1 DP.

[0199] Specifically, the stability and potency of DP mAb1 at 50 mg / mL stored at 5°C for 12 months were not affected by a ± 10% change in protein concentration, a ± 20% change in sucrose, L-proline and / or histidine concentration and / or a ± 50% change in polysorbate 80 concentration and / or a ± 0.3 unit change in pH.

[0200] The robustness of the mAb1 formulation was demonstrated by the PAR study. In general, the results of the pre-PAR and PAR studies supported that the variability in the compositions of the mAb1 formulation within the studied ranges would not adversely affect the stability of DP mAb1 under the recommended storage conditions (2 to 8°C).

[0201] Samples of FDS mAb1 at 50 mg / mL were stable after two freeze-thaw cycles (-30°C freeze-thaw and room temperature thaw). The stability of FDS mAb1 to freeze / thaw stress was not affected by a ±10% change in mAb1 concentration, a ±20% change in sucrose, and / or a ±0.3 unit change in pH relative to the control FDS mAb1 (50 mg / mL). The results of these freeze / thaw studies provide support that FDS mAb1 at 50 mg / mL can be frozen and thawed during the manufacture of DP mAb1 without adversely affecting FDS stability. Example 14: Containers

[0202] The mAb1 formulations were developed in glass vials (for distribution via intravenous infusion). The container for the mAb1 drug intended for further clinical development and Petition 870220073975, dated 08 / 17 / 2022, pp. 124 / 245 114 / 130 The product is also marketed as a pre-filled syringe, which is presented as a standalone syringe for self-injection or incorporated into an auto-injector device for self-administration. Example 15: Stability of mAb1 formulation in glass vials

[0203] Tables 32-35 summarize the stability of exemplary mAb1 formulations in 10 mL glass vials. Petition 870220073975, dated 08 / 17 / 2022, pages 125 / 245 Table 32: Stability study of the mAbl formulation stored at 2-8°C Formulation mAbl at 50 mg / mL, L-histidine at 10 mM, sucrose at 5% (w / v), L-proline at 1.5% (w / v) and polysorbate 80 at 0.2% (w / v), pH 6.0 Filling volume 5.5 mL Container / Seal 10 mL type 1 glass vials with a 20 mm chlorobutyl stopper 4432 / 50 coated with FluroTec® Assay Storage duration at 5°C (months) 0 1 3 6 9 12 18 24 Color and appearance Approved Approved Approved Approved Approved Approved Approved Turbidity (DO increase at 405 nm) 0.00 0.00 0.00 0.00 0.00 0.00 0.00 pH 6.0 6.0 6.0 6.0 6.0 6.0 6.0 Subvisible particulate analysis by IMF (N / mL) 2 to 10 pm 175 NR 62 772 NR 1730 NR > 10 pm 8 NR 3 144 NR 18 NR > 25 pm 0 NR 1 19 NR 4 NR % protein recovered by SE-UPLC 100 102 103 101 105 103 104 Purity by MCE-SDS Not reduced; % main peak 99.1 NR NR 99.1 NR 99.2 NR Reduced;% Heavy + Light Chain 100 NR NR 100 NR 100 NR Purity by SE-UPLC % HMW 0.3 0.3 0.3 0.4 0.4 0.4 0.4 % Monomers 99.1 99.3 99.3 99.3 99.2 99.2 99.2 % LMW 0.6 0.4 0.4 0.4 0.4 0.4 0.4 Charge Variant Analysis by CEX-UPLC % Acid 18.9 18.9 18.9 19.8 20.5 19.8 18.9 % Principal 53.5 53.3 53.4 54.7 54.3 54.0 52.9 % Basic 27.5 27.8 27.7 25.6 25.2 26.2 28.2 Load Variant Analysis by iCIEF % Acid 33.8 NR 34.3 34.2 NR 34.9 NR % Principal 54.8 NR 54.2 54.3 NR 54.3 NR % Basic 11.4 NR 11.4 11.4 NR 10.8 NR % Relative Potency (Bioassay) 92 NR NR 122 NR 112 NR; 115 / 130 Petition 870220073975, dated 08 / 17 / 2022, pp. 126 / 245 116 / 130 CEX, cation exchange; DS, medicinal substance; HMW, high molecular weight; iCIEF, capillary focusing isoelectric imaging; LMW, low molecular weight; IMF, microflow; NR, not mandatory; OD, optical density; PR, reversed phase; SE, size exclusion; UPLC, high-performance liquid chromatography Petition 870220073975, dated 08 / 17 / 2022, pp. 127 / 245 Table 33: Stability study of the mAbl formulation under accelerated and stress conditions. Formulation mAbl at 50 mg / mL, L-histidine at 10 mM, sucrose at 5% (w / v), L-proline at 1.5% (w / v) and polysorbate 80 at 0.2% (w / v), pH 6.0 Filling volume 5.5 mL Container / Seal 10 mL type 1 glass bottles with a 20 mm chlorobutyl stopper 4432 / 50 coated with FluroTec® Storage 25°C / 60% RH (months) Storage 40°C (days) Assay 0 1 3 6 7 14 28 Color and appearance Approved Approved Approved Approved Approved Approved Approved Turbidity (DO increase at 405 nm) 0.00 0.00 0.00 0.00 0.00 0.00 0.00 pH 6.0 6.0 6.0 6.0 6.0 6.0 6.0 Subvisible particulate analysis by MFI (# / mL) 2-10 pm 175 NR 326 2083 NR NR 288 > 10 pm 8 NR 4 109 NR NR 14 > 25 pm 0 NR 3 3 NR NR 1 % protein recovered by SE-UPLC 100 102 104 101 100 101 102 Purity by MCE-SDS Not reduced; % main peak 99.1 NR NR 98.8 NR NR 98.6 Reduced;% Heavy + Light Chain 100 NR NR 99.7 NR NR 99.7 Purity by SE-UPLC % HMW 0.3 0.4 0.5 0.6 0.5 0.8 1.2 % Monomers 99.1 99.1 99.1 99.0 98.7 98.5 98.1 % LMW 0.6 0.5 0.4 0.4 0.7 0.7 0.7 Charge Variant Analysis by CEX-UPLC % Acid 18.9 19.4 22.1 28.3 20.0 22.0 27.1 % Principal 53.5 52.7 51.2 48.6 51.9 50.2 46.7 % Basic 27.5 27.9 26.8 23.2 27.1 27.7 26.2 Analysis of charge variants per CIEF % Acid 33.8 NR NR 41.9 NR NR 46.4 % Principal 54.8 NR NR 46.5 NR NR 40.9 % Basic 11.4 NR NR 11.6 NR NR 12.7 % relative potency per bioassay 92 NR NR 91 NR NR 83; 117 / 130 CEX, cation exchange; DS, medicinal substance; HMW, high molecular weight; iCIEF, Petition 870220073975, dated 08 / 17 / 2022, pp. 128 / 245: isoelectric focusing imaging in capillaries; LMW, low molecular weight; IMF, microflow; NR, not mandatory; OD, optical density; PR, reversed phase; SE, size exclusion; UPLC, high-performance liquid chromatography. Table 34: Stability study of the mAbl formulation stored at 2-8°C Formulation mAbl at 50 mg / mL, L-histidine at 10 mM, sucrose at 5% (w / v), L-proline at 1.5% (w / v) and polysorbate 80 at 0.2% (w / v), pH 6.0 Filling volume 7.44 mL Container / Seal 10 mL type 1 glass bottles with a 20 mm chlorobutyl stopper 4432 / 50 coated with FluroTec® Test Duration of storage at 2-8°C (months 0 1 3 6 9 12 18 24 Color and appearance Approved Approved Approved Approved Approved Turbidity (DO increase at 405 nm) 0.00 0.00 0.00 0.00 0.00 pH 6.0 6.0 6.0 6.0 6.0 Subvisible particulate analysis by HIAC (N / mL) > 10 pm 3 NR 2 4 NR s 25 pm 0 NR 0 1 NR Subvisible particulate analysis by IMF (N / mL) 2 to 10 pm 183 NR 207 1044 NR > 10 pm 3 NR 7 19 NR s 25 pm 1 NR 3 5 NR % protein recovered by RP-UPLC 100 99 102 102 94 Purity by MCE-SDS Not reduced; % main peak 99.2 NR 99.4 99.2 NR Reduced; % heavy + light chain 100 NR 100 100 NR Purity by SE-UPLC % HMW 0.7 0.6 0.5 0.5 0.6% monomers 98.8 98.8 98.9 99.0 98.9% LMW 0.6 0.6 0.5 0.4 0.5 118 / 130 Petition 870220073975, dated 08 / 17 / 2022, pp. 129 / 245 Formulation mAbl at 50 mg / mL, L-histidine at 10 mM, sucrose at 5% (w / v), L-proline at 1.5% (w / v) and polysorbate 80 at 0.2% (w / v), pH 6.0 Filling volume 7.44 mL Container / Seal 10 mL type 1 glass vials with a 20 mm chlorobutyl stopper 4432 / 50 coated with FluroTec® Assay Storage duration at 2-8°C (months 0 1 3 6 9 12 18 24 Charge variant analysis by CEX-UPLC % Acid 21.2 21.2 22.3 19.6 21.5 % Principal 52.0 52.1 52.0 51.9 52.0 % Basic 26.8 26.8 25.7 28.6 26.6 Analysis of charge variants by iCIEF % Acid 34.3 NR 34.7 34.6 NR % Principal 52.1 NR 51.9 51.9 NR % Basic 13.7 NR 13.4 13.4 NR % relative potency (bioassay) 113 NR 105 128 NR CEX, cation exchange; DS, medicinal substance; HMW, high molecular weight; iCIEF, 119 / 130 Isoelectric focusing imaging in capillaries; LMW, low molecular weight; IMF, microflow; NR, not mandatory; OD, optical density; PR, reversed phase; SE, size exclusion; UPLC, high-performance liquid chromatography Petition 870220073975, dated 08 / 17 / 2022, pp. 130 / 245 Table 35: Stability study of the mAbl formulation under accelerated and stress conditions. Formulation mAbl at 50 mg / mL, L-histidine at 10 mM, sucrose at 5% (w / v), L-proline at 1.5% (w / v) and polysorbate 80 at 0.2% (w / v), pH 6.0 Filling volume 7.44 mL Container / Seal 10 mL type 1 glass bottles with a 20 mm chlorobutyl stopper 4432 / 50 coated with FluroTec® Storage 25°C / 60% RH (months) 40°C Storage (days) Assay 0 1 3 6 7 14 28 Color and appearance Approved Approved Approved Approved Approved Approved Approved Approved Turbidity (increase in DO at 405 nm) 0.00 0.00 0.00 0.00 0.00 0.00 0.00 pH 6.0 6.0 6.0 6.0 6.0 6.0 6.0 Subvisible particulate analysis by HIAC (N / mL) > 10 pm 3 NR 4 11 NR NR 5 s 25 pm 0 NR 0 2 NR NR 0 Subvisible particulate analysis by MFI (# / mL) 2-10 pm 183 NR 540 1439 NR NR 799 > 10 pm 3 NR 18 16 NR NR 92 s 25 pm 1 NR 1 1 NR NR 1 % protein recovered by SE-UPLC 100 100 101 101 101 99 100 Purity by MCE-SDS Not reduced; % peak main 99.2 NR 99.3 98.7 NR NR 98.6 Reduced;% Heavy + Light Chain 100 NR 100 100 NR NR 99.5 Purity by SE-UPLC % HMW 0.7 0.6 0.7 0.8 0.6 0.8 1.3 % Monomers 98.8 98.8 98.8 98.7 98.7 98.5 97.9 % LMW 0.6 0.7 0.6 0.6 0.7 0.8 0.8 Charge Variant Analysis by CEX-UPLC % Acid 21.2 21.7 25.3 24.4 20.5 23.2 25.8 % Principal 52.0 52.1 51.4 50.5 52.4 51.0 49.7; 120 / 130 Petition 870220073975, dated 08 / 17 / 2022, pp. 131 / 245 Formulation: mAb1 at 50 mg / mL, L-histidine at 10 mM, sucrose at 5% (w / v), L-proline at 1.5% (w / v) and polysorbate 80 at 0.2% (w / v), pH 6.0. Filling volume: 7.44 mL. Container / Seal: 10 mL Type 1 glass vials with a 20 mm chlorobutyl stopper. 4432 / 50 coated with FluroTec®. Storage: 25°C / 60% RH (months). Storage (days): 40°C. Assay: 0 1 3 6 7 14 28% Basic 26.8 26.2 23.3 25.1 27.0 25.9 24.6. Charge variant analysis by iCIEF: % Acid 34.3 NR 40.0 45.4 NR NR 46.9 % Principal 52.1 NR 47.1 43.7 NR NR 39.7 % Basic 13.7 NR 12.9 11.0 NR NR 13.3 % Relative potency per bioassay 113 NR 143 99 NR NR 88 CEX, cation exchange; DS, medicinal substance; HMW, high molecular weight; iCIEF, capillary focusing isoelectric imaging; LMW, low molecular weight; IMF, microflow; NR, not mandatory; OD, optical density; PR, reversed phase; SE, size exclusion; UPLC, high-performance liquid chromatography

[0204] It was found that the two formulations at different filling volumes are stable under stress (40°C / 75% RH) (data not shown). Example 16: Stability of the mAb1 formulation in pre-filled syringes

[0205] Tables 36-38 summarize the stability of high-concentration mAb1 formulations in pre-filled syringes. Table 36: Stability study of the mAb1 medicinal product in pre-filled syringes (PFS) stored at 5 °C 121 / 130 Petition 870220073975, dated 08 / 17 / 2022, pp. 132 / 245 Formulation mAbl at 175 mg / mL, histidine at 10 mM, sucrose at 5% (w / v), L-proline at 1.5% (w / v), polysorbate 80 at 0.2% (w / v), pH 6.0 Filling volume 1.2 mL Container / Seal 2.25 mL Nuova Ompi EZ Fill glass syringe with a 27G 1 / 2 thin-walled needle and FM30 needle protector closed with a FluroTec®-coated 4023 / 50 rubber plunger Storage duration at 5°C (months) Test 0 1 3 6 9 12 Color and appearance Approved Approved Approved Approved Approved Approved Turbidity (DO increase at 405 nm) 0.00 0.01 0.00 0.00 0.00 0.00 pH 6.0 6.0 6.0 6.0 6.0 6.0 Subvisible particulate analysis by HIAC (# / mL) > 10 pm 35 NR 17 59 NR 13 > 25 pm 0 NR 4 1 NR 1 Particulate analysis by MFI (particles / mL) 2 to 10 pm 21326 NR 8613 N / A NR N / A > 10 pm 82 NR 303 N / A NR N / A > 25 pm 3 NR 2 N / A NR N / A % protein recovered by RP-UPLC 100 96 97 100 98 100 Purity by MCE-SDS Not reduced; % peak main 97.6 NR NR 97.7 97.1 N / A Reduced;% Heavy + Light Chain 99.6 NR NR 99.8 99.8 N / A Purity by SE-UPLC % HMW 0.6 0.6 0.7 0.7 0.8 0.9 % Native 99.1 98.9 99.1 98.7 98.7 98.6 % LMW 0.3 0.5 0.2 0.5 0.5 0.5; 122 / 130 Petition 870220073975, dated 08 / 17 / 2022, pp. 133 / 245 Formulation mAbl at 175 mg / mL, histidine at 10 mM, sucrose at 5% (w / v), L-proline at 1.5% (w / v), polysorbate 80 at 0.2% (w / v), pH 6.0 Filling volume 1.2 mL Container / Seal 2.25 mL Nuova Ompi EZ Fill glass syringe with a 27G 1 / 2 thin-walled needle and FM30 needle protector closed with a FluroTec® coated 4023 / 50 rubber plunger Storage duration at 5°C (months) Assay 0 1 3 6 9 12 Charge variant analysis by CEX-UPLC % Acid 18.6 18.5 18.2 18.6 19.3 18.8 % Principal 53.4 54.8 54.3 53.2 53.5 55.5 % Basic 27.9 26.8 27.5 25.3 25.6 24.3 Analysis of charge variants by CIEF % Acid 30.2 NR 30.2 32.9 NR N / D % Principal 55.9 NR 55.9 53.4 NR N / D % Basic 13.9 NR 13.9 13.7 NR N / D % relative potency (bioassay) 87 NR NR 141 NR N / D 123 / 130 Petition 870220073975, dated 08 / 17 / 2022, pp. 134 / 245 Table 37: Stability test of mAbl drug in pre-filled syringe (PFS) stored under accelerated conditions Formulation mAbl at 175 mg / mL, histidine at 10 mM, sucrose at 5% (w / v), L-proline at 1.5% (w / v), polysorbate 80 at 0.2% (w / v), pH 6.0 Filling volume 1.2 mL Container / Seal Nuova Ompi EZ Fill 2.25 mL glass syringe with a 27G 1 / 2 thin-walled needle and FM30 needle protector closed with a FluroTec® coated 4023 / 50 rubber plunger Storage at 25°C / 60% RH (months) Storage at 40°C (days) Assay 0 1 3 6 7 14 28 Color and appearance Approved Approved Approved Approved Approved Approved Approved Turbidity (increase in DO at 405 nm) 0.00 0.00 0.00 0.00 0.00 0.00 0.00 pH 6.0 6.0 6.0 6.1 6.0 6.0 6.0 Subvisible particulate analysis by HIAC (# / mL) > 10 pm 3 NR 13 30 NR NR 14 s 25 pm 1 NR 1 1 NR NR 3 Particulate analysis by MFI (particles / mL) 2-10 pm 21326 NR 4605 N / D NR NR 3717 > 10 pm 82 NR 225 N / D NR NR 51 > 25 pm 3 NR 2 N / D NR NR 8 % protein recovered by RP-UPLC 100 98 98 100 100 100 98 Purity by MCE-SDS Not reduced;% Main Peak 97.6 NR 97.7 96.5 NR NR 96.8 Reduced; % Heavy + Light Chain 99.6 NR 99.2 99.7 NR NR 99.4 Purity by SE-UPLC % HMW 0.6 0.9 1.2 1.6 1.6 2.5 3.9 % Native 99.1 98.6 98.5 97.7 97.8 96.9 95.5; 124 / 130 Petition 870220073975, dated 08 / 17 / 2022, pages 135 / 245 Formulation mAbl at 175 mg / mL, histidine at 10 mM, sucrose at 5% (w / v), L-proline at 1.5% (w / v), polysorbate 80 at 0.2% (w / v), pH 6.0 Filling volume 1.2 mL Container / Seal Nuova Ompi EZ Fill 2.25 mL glass syringe with a 27G 1 / 2 thin-walled needle and FM30 needle protector closed with a FluroTec® coated 4023 / 50 rubber plunger Storage at 25°C / 60% RH (months) Storage at 40°C (days) Assay 0 1 3 6 7 14 28 % LMW 0.3 0.5 0.3 0.6 0.6 0.7 0.6 Load variant analysis by CEX-UPLC % Acid 18.6 19.0 21.3 25.5 20.0 22.1 24.3 % Principal 53.4 54.3 52.0 46.5 51.8 49.8 48.9 % Basic 27.9 26.8 26.6 28.1 28.3 28.1 26.8 Charge variant analysis by iCIEF % Acid 30.2 NR 36.8 44.9 NR NR 45.6 % Principal 55.9 NR 49.6 42.8 NR NR 40.5 % Basic 13.9 NR 13.6 12.3 NR NR 13.9 % relative potency by bioassay 87 NR NR 137 NR NR 83 125 / 130 Petition 870220073975, dated 08 / 17 / 2022, pp. 136 / 245 Table 38: Drug stability study of mAbl in pre-filled syringe (PFS) - Effect of stress conditions Formulation mAbl at 175 mg / mL, histidine at 10 mM, sucrose at 5% (w / v), L-proline at 1.5% (w / v), polysorbate 80 at 0.2% (w / v), pH 6.0 Filling volume 1.2 mL Container / Seal 2.25 mL Nuova Ompi EZ Fill glass syringe with a 27G 1 / 2 thin-walled needle and FM30 needle protector closed with a FluroTec® coated 4023 / 50 rubber plunger Agitation (min) Assay 0 60 120 Color and appearance Approved Approved Approved Turbidity (DO increase at 405 nm) 0.00 0.00 0.00 pH 6.0 6.0 6.0 Subvisible particulate analysis by HIAC (# / mL) > 10 pm 3 NR 5 > 25 pm 1 NR 5 Particulate analysis by MFI (particles / mL) 2 to 10 pm 21326 NR 13896 > 10 pm 82 NR 55 > 25 pm 3 NR 3 % protein recovered by RP-UPLC 100 100 100 Purity by MCE-SDS Not reduced; % main peak 97.6 NR 97.9 Reduced;% Heavy + Light Chain 99.6 NR 99.5 Purity by SE-UPLC % HMW 0.6 0.6 0.6 % Native 99.1 98.9 98.8 % LMW 0.3 0.6 0.6 Charge Variant Analysis by CEX-UPLC % Acid 18.6 18.8 19.0 % Principal 53.4 53.2 53.5 % Basic 27.9 28.1 27.5 Charge Variant Analysis by iCIEF % Acid 30.2 NR 30.4 % Principal 55.9 NR 55.5 % Basic 13.9 NR 14.1 % Relative Potency by Bioassay 87 NR 100; 126 / 130 Petition 870220073975, dated 08 / 17 / 2022, pp. 137 / 245 127 / 130

[0206] Example 17: Compatibility of mAbl formulations in intravenous (IV) administration devices

[0207] For compatibility assessment, the 50 mg / mL mAb1 formulation was added to a 100 mL IV administration bag containing 0.9% sodium chloride or 5% dextrose injection to evaluate whether mAb1 is stable when administered intravenously. To account for variability in patient weight, two mixing concentrations, 1.0 mg / mL mAb1 and 25 mg / mL mAb1, were examined in this study to reflect low and high dosage conditions. The following IV mixing components were used during the compatibility studies: • Drug Product: mAb1 anti-PD 50 mg / mL • Diluents: 0.9% Sodium Chloride for Injection, 5% Dextrose for Injection • IV Bags: Polyvinyl chloride (PVC) IV bags with di-(2-ethylhexyl) phthalate (DEHP) filled with 0.9% sodium chloride for injection; Polyvinyl chloride (PVC) IV bags with DEHP filled with 5% dextrose for injection; Polyolefin (PO) IV bags filled with 0.9% sodium chloride for injection; Polyolefin (PO) IV bags filled with 5% dextrose for injection; Polypropylene IV bags filled with 0.9% sodium chloride for injection; Polypropylene intravenous bottles filled with 0.9% sodium chloride for injection Petition 870220073975, dated 08 / 17 / 2022, pp. 138 / 245 128 / 130 • IV Pumps o Peristaltic pump o Fluid displacement pump • IV Infusion Sets o PVC IV set with DEHP o PVC IV set with dioctyl terephthalate (DEHT) o PVC IV set with trioctyl trimellitate (TOTM) o Polyethylene-coated PVC IV set o Polyurethane IV set • Filters o 0.2 μm polyethersulfone in-line filter o 1.2 μm polyethersulfone in-line filter o 5 μm polyethersulfone in-line filter o 15 μm polyethersulfone in-line filter

[0208] The PDs used in this study were GMP manufactured using a representative commercial PD manufacturing process. The IV administration bags containing the mixture were initially kept for 24 hours at 5°C; the bags were then incubated for at least 8 hours at 25°C. After these incubations, each infusion set was connected to the intravenous bag, primed with the mixture, and kept for 1 hour at room temperature. Each mixture was then pumped through the respective infusion sets at rates of 25 mL / h and 500 mL / h. Methods used to evaluate the compatibility of the mixture:

[0209] The compatibility of the mAb1 mixture with the materials used in the IV administration device was evaluated using the following tests: • Color and appearance by visual inspection • pH • Turbidity measured by the increase in optical density (OD) Petition 870220073975, dated 08 / 17 / 2022, pp. 139 / 245 129 / 130 at 405 nm • Analysis of subvisible particulates in the mixture by light obscuring (HIAC) • Protein concentration of mAb1 by reversed-phase high-performance liquid chromatography (RP-UPLC) • Purity by SE-UPLC • Analysis of charge variants by CEX-UPLC • Potency, by bioassay: The relative potency of each sample is determined using the bioassay and is defined as: (IC50 of the reference sample / IC50 of the sample) * 100%. The measured potency of the samples in relation to storage stability should be within 50-150% of the measured potency of the reference standard. Results and Conclusions:

[0210] The 50 mg / mL mAb1 formulation diluted in both 0.9% sodium chloride and 5% dextrose for injection to concentrations of 1.0 mg / mL or 25 mg / mL was physically and chemically stable under all conditions tested within the proposed dose ranges and administration conditions. These data support the following conclusions regarding the dose preparation and IV administration of DP mAb1: • The 0.9% Sodium Chloride injection and 5% Dextrose injection bags, made of PVC with DEHP, PO and polypropylene, are compatible with the IV administration of mAb1. • DP mAb1 can be diluted to concentrations as low as 1.0 mg / mL in PVC, PO, or polypropylene IV administration bags containing 0.9% sodium chloride or 5% dextrose for IV administration. • DP mAb1 can be diluted to concentrations as high as 25.0 mg / mL in PVC, PO, or polypropylene IV administration bags containing 0.9% sodium chloride or 5% dextrose. Petition 870220073975, dated 08 / 17 / 2022, pages 140 / 245 130 / 130 for administration IV. • The mAb1 mixture in 0.9% sodium chloride or 5% dextrose remained stable after incubation in a PVC, PO, or polypropylene IV administration bag for periods of up to 24 hours at 5°C and 8 hours at 25°C. The diluted mAb1 DP can be administered within 6 hours of preparation. • Diluted mAb1 can be administered using a standard infusion pump. • Diluted mAb1 can be administered with an infusion set made of PVC containing DEHP, PVC containing TOTM, polyethylene, or polyurethane. • The mAb1 is compatible with the use of a 0.2 μm - 5 μm in-line polyethersulfone filter. • Diluted mAb1 can be administered at a rate ranging from 25 to 500 mL / hour.

[0211] The scope of the present invention should not be limited by the specific embodiments described herein. In fact, various modifications of the invention, in addition to those described herein, will become apparent to those skilled in the art from the preceding description and the accompanying figures. Such modifications should fall within the scope of the appended claims. Petition 870220073975, dated 08 / 17 / 2022, pp. 141 / 245

Claims

1 / 9 CLAIMS 1. A stable liquid pharmaceutical formulation, characterized in that it comprises: (a) an antibody that specifically binds to human programmed death protein-1 (PD-1), wherein the antibody comprises three heavy chain complementarity-determining regions (CDRs) (HCDR1, HCDR2 and HCDR3) contained within a heavy chain variable region (HCVR) of SEQ ID NO: 1 and three light chain CDRs (LCDR1, LCDR2 and LCDR3) contained within a light chain variable region (LCVR) of SEQ ID NO: 2; (b) histidine buffer; (c) polysorbate; (d) sucrose; and (e) proline; wherein said stable liquid pharmaceutical formulation has a pH of 6.0 ± 0.

3.

2. Pharmaceutical formulation, according to claim 1, characterized in that the antibody concentration is from 5 mg / mL ± 0.75 mg / mL to 250 mg / mL ± 37.5 mg / mL.

3. Pharmaceutical formulation, according to claim 2, characterized in that the antibody concentration is 25 mg / mL ± 3.75 mg / mL.

4. Pharmaceutical formulation, according to claim 2, characterized in that the antibody concentration is 50 mg / mL ± 7.5 mg / mL.

5. Pharmaceutical formulation, according to claim 2, characterized in that the antibody concentration is 150 mg / mL ± 22.5 mg / mL.

6. Pharmaceutical formulation, according to claim 2, characterized in that the antibody concentration is 175 Petition 870220073975, dated 08 / 17 / 2022, page 142 / 245 2 / 9 mg / mL ± 26.25 mg / mL.

7. Pharmaceutical formulation, according to any one of claims 1 to 6, characterized in that the concentration of the histidine buffer is from 5 mM ± 1 mM to 20 mM ± 4 mM.

8. Pharmaceutical formulation according to claim 7, characterized in that the concentration of the histidine buffer is 10 mM ± 2 mM.

9. Pharmaceutical formulation, according to claim 7 or 8, characterized in that the histidine buffer comprises L-histidine and L-histidine monohydrochloride monohydrate.

10. Pharmaceutical formulation according to claim 9, characterized in that the histidine buffer is prepared from 4.8 mM ± 0.96 mM of L-histidine and 5.2 mM ± 1.04 mM of L-histidine monohydrochloride monohydrate.

11. Pharmaceutical formulation, according to any one of claims 1 to 10, characterized in that the polysorbate concentration is from 0.01% ± 0.005% to 0.5% ± 0.25% w / v.

12. Pharmaceutical formulation, according to claim 11, characterized in that the polysorbate concentration is 0.1% ± 0.05% w / v.

13. Pharmaceutical formulation, according to claim 11, characterized in that the polysorbate concentration is 0.2% ± 0.1% w / v.

14. Pharmaceutical formulation, according to any one of claims 11 to 13, characterized in that the polysorbate is polysorbate 80.

15. Pharmaceutical formulation, according to any one of claims 1 to 14, characterized in that the sucrose concentration is from 1% ± 0.2% to 20% ± 4% w / v. Petition 870220073975, dated 17 / 08 / 2022, pp. 143 / 245 3 / 9 16. Pharmaceutical formulation, according to claim 15, characterized in that the sucrose concentration is from 1% ± 0.2% to 10% ± 2% w / v.

17. Pharmaceutical formulation, according to claim 16, characterized in that the sucrose concentration is 5% ± 1% w / v.

18. Pharmaceutical formulation, according to any one of claims 1 to 17, characterized in that the proline concentration is from 1% ± 0.2% to 5% ± 1% w / v.

19. Pharmaceutical formulation, according to claim 18, characterized in that the proline concentration is 1.5% ± 0.3% w / v.

20. Pharmaceutical formulation, according to any one of claims 1 to 19, characterized in that it has a viscosity of less than 20 cP at 25°C.

21. Pharmaceutical formulation according to claim 1, characterized in that it comprises: (a) 175 mg / mL ± 26.25 mg / mL of antibody, (b) 10 mM ± 2 mM histidine buffer, (c) 0.2% ± 0.1% w / v polysorbate, (d) 5% ± 1% w / v sucrose, and (e) 1.5% ± 0.3% w / v proline; in water, wherein: (i) > 90% of the antibodies have a molecular weight of 143 kDa ± 1 kDa; and / or (ii) the pharmaceutical formulation has a viscosity of less than 20 cP at 25°C.

22. Pharmaceutical formulation, according to claim 21, characterized in that the histidine buffer is prepared from 4.8 mM ± 0.96 mM of L-histidine and 5.2 mM ± 1.04 mM of L-histidine monohydrochloride monohydrate.

23. Pharmaceutical formulation according to claim 21, characterized in that it comprises: (a) 175 mg / mL of antibody, (b) 0.74 mg / mL of L-histidine, (c) 1.1 mg / mL of L-histidine monohydrochloride monohydrate, (d) 2 mg / mL of polysorbate, (e) 50 mg / mL of sucrose, and (f) 15 mg / mL of proline.

24. Pharmaceutical formulation according to claim 1, characterized in that it comprises: (a) 150 mg / mL ± 22.5 mg / mL of antibody, (b) 10 mM ± 2 mM histidine buffer, (c) 0.2% ± 0.1% w / v polysorbate, (d) 5% ± 1% w / v sucrose, and (e) 1.5% ± 0.3% w / v proline, in water, wherein: (i) > 90% of the antibodies have a molecular weight of 143 kDa ± 1 kDa; and / or (ii) the pharmaceutical formulation has a viscosity of less than 20 cP at 25°C.

25. Formulação farmacêutica, de acordo com a reivindicação 24, caracterizada pelo fato de que o tampão de histidina é preparado a partir de 4,8 mM ± 0,96 mM L-histidina e 5,2 mM ± 1,04 mM monocloridrato de L-histidina monoidratado.

26. Pharmaceutical formulation, according to claim 24, characterized in that it comprises: Petition 870220073975, dated 17 / 08 / 2022, page 145 / 245 5 / 9 (a) 150 mg / mL of antibody, (b) 0.74 mg / mL of L-histidine, (c) 1.1 mg / mL of L-histidine monohydrochloride monohydrate, (d) 2 mg / mL of polysorbate, (e) 50 mg / mL of sucrose, and (f) 15 mg / mL of proline.

27. Pharmaceutical formulation according to claim 1, characterized in that it comprises: (a) 50 mg / mL ± 7.5 mg / mL of antibody, (b) 10 mM ± 2 mM histidine buffer, (c) 0.2% ± 0.1% w / v polysorbate, (d) 5% ± 1% w / v sucrose, and (e) 1.5% ± 0.3% w / v proline, in water, wherein: (i) > 90% of the antibodies have a molecular weight of 143 kDa ± 1 kDa; and / or (ii) the pharmaceutical formulation has a viscosity of less than 20 cP at 25°C.

28. Pharmaceutical formulation according to claim 27, characterized in that the histidine buffer is prepared from 4.8 mM ± 0.96 mM L-histidine and 5.2 mM ± 1.04 mM L-histidine monohydrochloride monohydrate.

29. Pharmaceutical formulation, according to claim 27, characterized in that it comprises: (a) 50 mg / mL of antibody, (b) 0.74 mg / mL of L-histidine, (c) 1.1 mg / mL of L-histidine monohydrochloride monohydrate, Petition 870220073975, dated 17 / 08 / 2022, page 146 / 245 6 / 9 (d) 2 mg / mL of polysorbate, (e) 50 mg / mL of sucrose, and (f) 15 mg / mL of proline.

30. Pharmaceutical formulation according to claim 1, characterized in that it comprises: (a) 25 mg / mL ± 3.75 mg / mL of antibody, (b) 10 mM ± 2 mM histidine buffer, (c) 0.2% ± 0.1% w / v polysorbate, (d) 5% ± 1% w / v sucrose, and (e) 1.5% ± 0.3% w / v proline, in water, wherein: (i) > 90% of the antibodies have a molecular weight of 143 kDa ± 1 kDa; and / or (ii) the pharmaceutical formulation has a viscosity of less than 20 cP at 25°C.

31. Pharmaceutical formulation according to claim 30, characterized in that the histidine buffer is prepared from 4.8 mM ± 0.96 mM L-histidine and 5.2 mM ± 1.04 mM L-histidine monohydrochloride monohydrate.

32. Pharmaceutical formulation according to claim 30, characterized in that it comprises: (a) 25 mg / mL of antibody, (b) 0.74 mg / mL of L-histidine, (c) 1.1 mg / mL of L-histidine monohydrochloride monohydrate, (d) 2 mg / mL of polysorbate, (e) 50 mg / mL of sucrose, and (f) 15 mg / mL of proline.

33. Pharmaceutical formulation, according to any one of claims 1 to 32, characterized in that the antibody comprises an HCDR1 of SEQ ID NO: 3, an HCDR2 of SEQ ID NO: 4, an HCDR3 of SEQ ID NO: 5, an LCDR1 of SEQ ID NO: 6, an LCDR2 of SEQ ID NO: 7 and an LCDR3 of SEQ ID NO:

8.

34. Pharmaceutical formulation, according to claim 33, characterized in that the antibody comprises an HCVR with SEQ ID NO: 1 and an LCVR with SEQ ID NO:

2.

35. Pharmaceutical formulation, according to any one of claims 1 to 34, characterized in that the antibody comprises an HCVR that has 90% sequence identity with SEQ ID NO:

1.

36. Pharmaceutical formulation, according to any one of claims 1 to 34, characterized in that the antibody comprises an LCVR that has 90% sequence identity with SEQ ID NO:

2.

37. Pharmaceutical formulation, according to any one of claims 1 to 34, characterized in that the antibody comprises an HCVR that has 90% sequence identity with SEQ ID NO: 1 and an LCVR that has 90% sequence identity with SEQ ID NO:

2.

38. Pharmaceutical formulation, according to any one of claims 1 to 34, characterized in that the antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 9 and 11.

39. Pharmaceutical formulation, according to any one of claims 1 to 34, characterized in that the antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence of SEQ ID NO:

9.

40. Pharmaceutical formulation, according to any one of claims 1 to 34, characterized in that the antibody comprises a heavy chain and a light chain, the heavy chain comprising an amino acid sequence of SEQ ID NO:

11.

41. Pharmaceutical formulation, according to any one of claims 1 to 34, characterized in that the antibody comprises a heavy chain and a light chain, wherein the light chain comprises the amino acid sequence of SEQ ID NO:

10.

42. Pharmaceutical formulation, according to any one of claims 1 to 34, characterized in that the antibody comprises a heavy chain / light chain comprising amino acid sequences selected from the group consisting of SEQ ID NOs: 9 / 10 and 11 / 10.

43. Pharmaceutical formulation, according to any one of claims 1 to 34, characterized in that the antibody comprises a heavy chain / light chain comprising amino acid sequences of SEQ ID NOs: 9 / 10.

44. Pharmaceutical formulation, according to any one of claims 1 to 34, characterized in that the antibody comprises a heavy chain / light chain comprising amino acid sequences of SEQ ID NOs: 11 / 10.

45. Pharmaceutical formulation, according to any one of claims 1 to 44, characterized in that it is for subcutaneous or intravenous administration.

46. ​​Pharmaceutical formulation, according to any one of claims 1 to 45, characterized in that it is contained in a container.

47. Pharmaceutical formulation according to claim 46, characterized in that the container is a bottle.

48. Pharmaceutical formulation, according to claim Petition 870220073975, dated 08 / 17 / 2022, page 149 / 245 9 / 9 47, characterized by the fact that the bottle is a Type 1 transparent glass bottle of 10 mL.

49. Pharmaceutical formulation according to claim 46, characterized in that the container is a syringe.

50. Pharmaceutical formulation, according to claim 49, characterized in that the syringe is made of glass with a low tungsten content.

51. Pharmaceutical formulation according to claim 46, characterized in that the container is a pre-filled syringe.

52. Pharmaceutical formulation, according to claim 46, characterized in that it is contained in an autoinjector.

53. Kit, characterized in that it comprises a pharmaceutical formulation, as defined in any one of claims 1 to 45, a container and instructions.

54. Kit according to claim 53, characterized in that the container is a glass bottle.

55. Kit according to claim 53, characterized in that the container is a pre-filled syringe.

56. Kit, according to claim 53, characterized in that the container is an auto-injector. Petition 870220073975, dated 17 / 08 / 2022, pp. 150 / 245

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