Liquid formulation of an IL-22R antibody

Abstract

The present invention relates to an aqueous liquid antibody formulation containing an antibody that is suitable for injection in terms of stability, osmolality, viscosity, and syringe performance.

Description

Background Art

[0001] The present disclosure relates to aqueous liquid antibody formulations and other protein formulations that are suitable for injection in terms of stability, osmolality, viscosity, and syringe performance. Antibodies and other proteins can be administered to patients via subcutaneous injection. To ensure patient convenience, subcutaneous dosage forms are desirably free of injection pain or difficulty, and thus the formulation is preferably isotonic and has a sufficiently high concentration of active ingredient to achieve the desired clinical dose and desired clinical outcome while being administered in a relatively small injection volume. An increase in protein concentration is associated with an exponential increase in viscosity, which results in an increase in manufacturing risk, an increase in the risk associated with identifying the optimal device and needle solution, a decrease in injectability with a thin needle, and a potential decrease in patient convenience during injection. Furthermore, the formulation must be stable per se and provide a sufficient stabilizing environment for the protein / antibody to avoid structural degradation and protein aggregation, maintain the desired clinical effect of the formulation after storage, and provide an acceptable shelf life for the formulation.

[0002] Improvement of the storage stability of proteins can be achieved by freeze drying (lyophilization / freeze drying) which removes water (sublimation), changes the formulation from an aqueous formulation of the protein to a solid, and in principle results in a water-free matrix consisting of the protein and additives. However, such freeze-dried formulations require a reconstitution step prior to injection and are not suitable for prefilled syringes or autoinjectors. Thus, liquid formulations are preferred for patient convenience.

[0003] The present invention provides a thermally stable formulation of a liquid IL-22R antibody useful for treating skin conditions such as atopic dermatitis. The present invention also discloses a stable high-concentration formulation that allows for a small injection volume or a higher dose.

[0004] The use of stable liquid formulations is advantageous in the clinical setting and for patient compliance. For example, it is in contrast to lyophilized formulations that need to be reconstituted before use. Stable liquid formulations can be used in prefilled syringes or autoinjectors. High-concentration formulations of antibodies may be desirable for reducing the injection volume, particularly for formulations intended for subcutaneous administration. However, high-concentration antibody formulations often have problems due to, for example, insufficient stability resulting from protein aggregation and viscosity exceeding the threshold for simple manufacture and injection.

[0005] The present invention provides a formulation that solves the above problems.

Summary of the Invention

[0006] The present invention provides the following embodiments, all of which should be understood as independent embodiments or as embodiments dependent on any of the other embodiments listed.

[0007] A liquid pharmaceutical formulation comprising an IL-22R antibody at a concentration of 150 ± 15 mg / mL to 225 ± 25 mg / mL, comprising the following: One or more disaccharides, One or more amino acids, Optionally, a surfactant further comprising, having a pH of 5.5 to 6.5 and an osmolality of 280 to 450 mOsm / kg, a pharmaceutical formulation.

[0008] A liquid pharmaceutical formulation comprising an IL-22R antibody at a concentration of 150 ± 15 mg / mL to 200 ± 25 mg / mL, comprising the following: One or more disaccharides, One or more amino acids, Optionally, a surfactant further comprising, having a pH of 5.5 to 6.5 and an osmolality of 280 to 450 mOsm / kg, a pharmaceutical formulation.

[0009] A liquid pharmaceutical formulation comprising an IL-22R antibody at a concentration of 150 ± 15 mg / mL to 200 ± 25 mg / mL, comprising the following: One or more disaccharides, One or more amino acids, A buffer, An antioxidant, Optionally, a viscosity reducing agent, Optionally, a surfactant Further comprising, having a pH of 5.5 to 6.5 and an osmolality of 280 to 450 mOsm / kg, a pharmaceutical preparation.

[0010] A liquid pharmaceutical preparation comprising an IL-22R antibody at a concentration of 150 ± 15 mg / mL to 200 ± 25 mg / mL, the following: One or more disaccharides at a total concentration of 60 to 260 mM, One or more amino acids at a total concentration of 40 to 140 mM, Optionally, a surfactant Further comprising, having a pH of 5.5 to 6.5 and an osmolality of 280 to 450 mOsm / kg, a pharmaceutical preparation.

[0011] A liquid pharmaceutical preparation comprising an IL-22R antibody at a concentration of 150 ± 15 mg / mL to 200 ± 25 mg / mL, the following: One or more disaccharides at a total concentration of 60 to 260 mM, One or more amino acids at a total concentration of 40 to 140 mM, An antioxidant, A buffer, Optionally, a viscosity reducing agent, Optionally, a surfactant Further comprising, having a pH of 5.5 to 6.5 and an osmolality of 280 to 450 mOsm / kg, a pharmaceutical preparation.

[0012] A liquid pharmaceutical preparation comprising an IL-22R antibody at a concentration of 150 ± 15 mg / mL to 200 ± 25 mg / mL, the following: One or more disaccharides at a total concentration of 60 to 260 mM, One or more amino acids at a total concentration of 40 to 140 mM selected from the group consisting of glycine, proline, lysine, glutamic acid, methionine, arginine, aspartic acid and histidine, Optionally, a surfactant A pharmaceutical preparation that further contains it, has a pH of 5.5 to 6.5, and an osmolality of 280 to 450 mOsm / kg.

[0013] A liquid pharmaceutical preparation containing an IL-22R antibody at a concentration of 150 ± 15 mg / mL to 200 ± 25 mg / mL, comprising the following: One or more disaccharides at a total concentration of 60 to 260 mM, One or more amino acids at a total concentration of 40 to 140 mM selected from the group consisting of glycine, proline, lysine, glutamic acid, methionine, arginine, aspartic acid, and histidine, wherein the amino acids function as stabilizers, antioxidants, viscosity reducing agents, and buffering agents. Optionally, a surfactant A pharmaceutical preparation that further contains it, has a pH of 5.5 to 6.5, and an osmolality of 280 to 450 mOsm / kg.

[0014] A stable liquid pharmaceutical preparation containing an IL-22R antibody at a concentration of 150 ± 15 mg / mL to 200 ± 25 mg / mL, comprising the following: One or more disaccharides at a total concentration of 60 to 260 mM, One or more amino acids at a total concentration of 40 to 140 mM selected from the group consisting of glycine, proline, lysine, glutamic acid, methionine, arginine, aspartic acid, and histidine, Proline and / or glycine is a stabilizer, Methionine is an antioxidant, Arginine is a viscosity reducing agent, Histidine is a buffering agent, amino acids, and Optionally, a surfactant A pharmaceutical preparation that further contains it, has a pH of 5.5 to 6.5, and an osmolality of 280 to 450 mOsm / kg.

[0015] A stable liquid pharmaceutical preparation containing an IL-22R antibody at a concentration of 150 ± 15 mg / mL to 200 ± 25 mg / mL, comprising the following: One or more disaccharides at a total concentration of 60 to 260 mM, One or more amino acids selected from the group consisting of glycine, proline, lysine, glutamic acid, methionine, arginine, aspartic acid and histidine, having a total concentration of 40 to 140 mM, wherein proline and / or glycine is a stabilizer, methionine is an antioxidant, arginine is a viscosity reducing agent, histidine is a buffer, and optionally, a surfactant further comprising, having a pH of 5.5 to 6.5 and an osmolality of 280 to 450 mOsm / kg, a pharmaceutical preparation.

[0016] A stable liquid pharmaceutical preparation containing an IL-22R antibody at a concentration of 150 ± 15 mg / mL to 200 ± 25 mg / mL, comprising: One or more disaccharides having a total concentration of 60 to 260 mM, proline and / or glycine is present at a concentration of 0 to 80 mM, methionine is present at a concentration of 5 to 30 mM, arginine is present at a concentration of 0 to 100 mM, histidine is present at a concentration of 0 to 30 mM, and optionally, a surfactant further comprising, having a pH of 5.5 to 6.5 and an osmolality of 280 to 450 mOsm / kg, a pharmaceutical preparation.

[0017] A stable liquid pharmaceutical preparation containing an IL-22R antibody at a concentration of 150 ± 15 mg / mL to 225 ± 25 mg / mL, comprising: One or more disaccharides having a total concentration of 60 to 260 mM, glycine is present at a concentration of 0 to 80 mM, methionine is present at a concentration of 5 to 30 mM, arginine is present at a concentration of 0 to 100 mM, histidine is present at a concentration of 0 to 30 mM, and optionally, a surfactant A pharmaceutical preparation that further contains [substance], has a pH of 5.5 to 6.5, and an osmolality of 280 to 450 mOsm / kg.

[0018] A stable liquid pharmaceutical preparation containing an IL-22R antibody at a concentration of 150 ± 15 mg / mL to 200 ± 25 mg / mL, and having the following: One or more disaccharides at a total concentration of 60 to 260 mM, Glycine is present at a concentration of 0 to 80 mM, Methionine is present at a concentration of 5 to 30 mM, Arginine is present at a concentration of 0 to 100 mM, Histidine is present at a concentration of 0 to 30 mM, and Optionally, a surfactant A pharmaceutical preparation that further contains [substance], has a pH of 5.5 to 6.5, and an osmolality of 280 to 450 mOsm / kg.

[0019] The liquid pharmaceutical preparation according to the above embodiment, having a viscosity of less than 25 cP at 20 to 25°C.

[0020] The liquid pharmaceutical preparation according to the above embodiment, having a viscosity of less than 20 cP at 20 to 25°C.

[0021] The liquid pharmaceutical preparation according to any of the above embodiments, which is stable at 5°C for at least 3 years while maintaining the high molecular weight product at less than 5%.

[0022] The liquid pharmaceutical preparation according to any of the above embodiments, which is stable at 5°C for at least 2 years while maintaining the high molecular weight product at less than 5%.

[0023] The liquid pharmaceutical preparation according to any of the above embodiments, wherein the preparation contains a histidine buffer.

[0024] The liquid pharmaceutical preparation according to the above embodiment, wherein histidine is present at a concentration of about 10 to 30 mM.

[0025] The liquid pharmaceutical preparation according to the above embodiment, wherein histidine is present at a concentration of about 20 mM.

[0026] The liquid pharmaceutical preparation according to any one of the above embodiments, wherein the disaccharide is present at a concentration of about 80 to 240 mM.

[0027] The liquid pharmaceutical preparation according to any one of the above embodiments, wherein the disaccharide is present at a concentration of about 100 to 220 mM.

[0028] The liquid pharmaceutical preparation according to any one of the above embodiments, wherein the disaccharide is present at a concentration of about 120 to 200 mM.

[0029] The liquid pharmaceutical preparation according to any one of the above embodiments, wherein the disaccharide is present at a concentration of about 140 to 180 mM.

[0030] The liquid pharmaceutical preparation according to any one of the above embodiments, wherein the disaccharide is present at a concentration of about 60 to 120 mM.

[0031] The liquid pharmaceutical preparation according to any one of the above embodiments, wherein the disaccharide is present at a concentration of about 80 to 110 mM.

[0032] The liquid pharmaceutical preparation according to any one of the above embodiments, wherein the disaccharide is present at a concentration of about 100 to 180 mM.

[0033] The liquid pharmaceutical preparation according to any one of the above embodiments, wherein the disaccharide is trehalose or sucrose.

[0034] The liquid pharmaceutical preparation according to the above embodiment, wherein the disaccharide is trehalose.

[0035] The liquid pharmaceutical preparation according to any one of the above embodiments, wherein the amino acid is selected from the group consisting of glycine, proline, lysine, glutamic acid, methionine, arginine, aspartic acid and histidine.

[0036] The liquid pharmaceutical preparation according to the above embodiment, wherein the amino acids are glycine, methionine, arginine and histidine.

[0037] The liquid pharmaceutical preparation according to any one of the above embodiments, wherein the viscosity reducing agent is arginine.

[0038] The liquid pharmaceutical preparation according to any one of the above embodiments, wherein the antioxidant is methionine.

[0039] The liquid pharmaceutical preparation according to any one of the above embodiments, wherein the antioxidant methionine is present at a concentration of 10 to 30 mM.

[0040] The liquid pharmaceutical preparation according to any one of the above embodiments, wherein the antioxidant methionine is present at a concentration of 20 mM.

[0041] The liquid pharmaceutical preparation according to the above embodiment, wherein the surfactant is present at a concentration of 0.01 to 0.08% (w / w), 0.01 to 0.06% (w / w), 0.01 to 0.04% (w / w), 0.01 to 0.03%, 0.01 to 0.02% (w / w) or 0.02% (w / w).

[0042] The liquid pharmaceutical preparation according to any one of the above embodiments, wherein the surfactant is polysorbate 20, polysorbate 80 or poloxamer 188 at a total concentration of 0.01 to 0.03% (w / w).

[0043] An IL-22R antibody at a concentration of 150 ± 15 mg / mL, and One or more disaccharides at a total concentration of 180 to 260 mM, One or more amino acids at a total concentration of 40 to 120 mM, An antioxidant at a concentration of 5 to 30 mM, A viscosity reducing agent at a concentration of 0 to 100 mM, Optionally, a surfactant, A histidine buffer at a concentration of 20 mM Containing, having a pH of 5.6 to 6.5 and an osmolality of 280 to 450 mOsm / kg, the liquid pharmaceutical preparation according to any one of the above embodiments.

[0044] An IL-22R antibody at a concentration of 150 ± 15 mg / mL, and Trehalose at a total concentration of 180 to 260 mM, Glycine with a total concentration of 0 to 80 mM, Methionine with a concentration of 5 to 30 mM, Tween 20 with a total concentration of 0.01 to 0.03% (w / w), Histidine buffer with a concentration of 20 mM The liquid pharmaceutical preparation according to the above embodiment, which contains the above components, has a pH of 5.6 to 6.5, and an osmolality of 280 to 450 mOsm / kg.

[0045] An IL-22R antibody with a concentration of 150 ± 15 mg / mL, and Sucrose with a total concentration of 180 to 260 mM, Glycine with a total concentration of 0 to 80 mM, Methionine with a concentration of 5 to 30 mM, Tween 20 with a total concentration of 0.01 to 0.04% (w / w), Histidine buffer with a concentration of 20 mM The liquid pharmaceutical preparation according to the above embodiment, which contains the above components, has a pH of 5.6 to 6.5, and an osmotic pressure suitable for subcutaneous (SC) administration with an osmolality of 280 to 450 mOsm / kg.

[0046] An IL-22R antibody with a concentration of 150 ± 15 mg / mL, and Trehalose with a total concentration of about 180 mM, Glycine with a total concentration of about 80 mM, Methionine with a concentration of about 20 mM, Tween 20 with a total concentration of about 0.01 to 0.03% (w / w), Histidine buffer with a concentration of about 20 mM The liquid pharmaceutical preparation according to any of the above embodiments, which contains the above components and has a pH of 5.6 to 6.5.

[0047] Containing an IL-22R antibody with a concentration of 225 ± 25 mg / mL, and the following: Trehalose with a total concentration of 60 to 100 mM, Methionine with a concentration of 5 to 30 mM, Viscosity reducing agent with a concentration of 60 to 100 mM, Glycine with a concentration of 0 to 80 mM, Optionally, it further contains a surfactant The liquid pharmaceutical preparation according to any one of the above embodiments, which further contains , has a pH of 5.6 to 6.5, and an osmotic pressure of 280 to 450 mOsm / kg.

[0048] It contains an IL-22R antibody at a concentration of 200 ± 25 mg / mL and the following: Trehalose at a total concentration of 60 to 100 mM, Methionine at a concentration of 5 to 30 mM, A viscosity reducing agent at a concentration of 60 to 100 mM, Glycine at a concentration of 0 to 80 mM, Optionally, it further contains a surfactant The liquid pharmaceutical preparation according to any one of the above embodiments, which further contains , has a pH of 5.6 to 6.5, and an osmotic pressure of 280 to 450 mOsm / kg.

[0049] It contains an IL-22R antibody at a concentration of 200 ± 25 mg / mL and the following: Trehalose at a total concentration of 80 to 100 mM, Methionine at a concentration of 5 to 30 mM, Arginine at a concentration of 50 to 100 mM, Optionally, a surfactant at a total concentration of 0.01 to 0.03% (w / w), A histidine buffer at a concentration of 20 mM The liquid pharmaceutical preparation according to the above embodiment, which further contains , has a pH of 5.5 to 6.5, and an osmotic pressure of 280 to 450 mOsm / kg.

[0050] It contains an IL-22R antibody at a concentration of 200 ± 25 mg / mL and the following: Approximately 100 mM total concentration of trehalose, Approximately 20 mM concentration of methionine, Approximately 80 mM concentration of arginine, Approximately 0.02% (w / w) total concentration of polysorbate 20, Approximately 20 mM concentration of histidine buffer It further contains and has a pH of 5.5 to 6.5, The liquid pharmaceutical preparation according to any one of the above embodiments.

[0051] The liquid pharmaceutical preparation according to any one of the above embodiments, which is stable at 5°C for about 3 years.

[0052] The liquid pharmaceutical preparation according to any one of the above embodiments, which is stable at 5°C for about 2 years.

BEST MODE FOR CARRYING OUT THE INVENTION

[0053] The term "about" or "approximately" means within an acceptable error range of a specific value determined by those skilled in the art, which depends in part on how the value is measured or determined, i.e., the limitations of the measurement system. For example, "about" can mean within an acceptable standard deviation according to the practice in the art. Alternatively, "about" can mean a range up to ±20% of a given value, preferably up to ±10%, more preferably up to ±5%, and even more preferably up to ±1%. When a specific value is recited in the present application and the claims, unless otherwise stated, the term "about" is implied and in this context means within an acceptable error range of the specific value.

[0054] As used in this specification and the claims, the indefinite articles "a" and "an" should be understood to mean "at least one" unless expressly stated to the contrary. The term "and / or" as used in this specification and the claims should be understood to mean "either or both" of the elements so joined, i.e., elements that may be present conjunctively in some cases and disjunctively in other cases. A plurality of elements listed using "and / or" should likewise be construed as "one or more" of the elements so joined. Other elements other than those specifically identified by the "and / or" clause may be present whether or not they are related to those specifically identified elements. Thus, by way of non-limiting example, a reference to "A and / or B" when used with an open-ended phrase such as "comprising" may, in one embodiment, refer to only A (optionally including elements other than B), in another embodiment, only B (optionally including elements other than A), and in yet another embodiment, both A and B (optionally including other elements). The term "or" as used in this specification and the claims should be understood to have the same meaning as "and / or" as defined above. For example, when separating items in a list, "or" or "and / or" is inclusive, i.e., it includes at least one of some elements or a list of elements, but also includes two or more, and optionally also includes further unlisted items. Only terms expressly indicated to the contrary, such as "only one of" or "exactly one of", or when used in the claims, "consisting of", refer to including exactly one element of some elements or a list of elements. Generally, the term "or" as used in this specification should be construed as indicating an exclusive alternative (i.e., "either one or the other but not both") only when preceded by exclusive terms such as "either", "only one of", "only one of only", or "exactly one of". "Consisting essentially of", when used in the claims, has its ordinary meaning as used in the field of patent law.

[0055] As used in this specification and the claims, the term "one or more" means, in reference to a list of one or more elements, at least one element selected from any one or more of the elements in the list of elements, but does not necessarily include at least one of every element specifically listed in the list of elements, and is not intended to exclude any combination of elements in the list of elements. It should be understood that this definition also allows for the possibility that elements other than those specifically identified in the list of elements referred to by the term "one or more" may optionally be present, whether or not they are related to those specifically identified elements. Thus, by way of non-limiting example, "one or more of A and B" (or equivalently, "one or more of A or B", or equivalently "one or more A and / or B") may, in one embodiment, refer to at least one A, including optionally two or more, where B is absent (and optionally including elements other than B), and in another embodiment, may refer to at least one B, including optionally two or more, where A is absent (and optionally including elements other than A). It should be understood that, unless expressly stated to the contrary, in any method claimed herein that includes two or more steps or acts, the order of the method steps or acts is not necessarily limited to the order in which the method steps or acts are recited.

[0056] As used in this specification, the term "substantially" refers to a qualitative state indicating the overall or approximate degree of a characteristic or property of an object. Those skilled in the art of biology understand that it is rare, if ever, for biological and chemical phenomena to achieve or avoid absolute results. Thus, the term "substantially" is used herein to capture the potential lack of perfection inherent in many biological and chemical phenomena.

[0057] IL-22R (also known as IL-22R1 and IL-22RA) is a type II cytokine receptor that is selectively expressed in the skin and epithelial cells. This receptor mediates signal transduction via three cytokines, interleukin 22 (IL-22), interleukin 20 (IL-20), and interleukin 24 (IL-24). Cytokine signal transduction requires the formation of a heterodimeric complex on the cell surface. IL-22 binds to and signals through a complex consisting of IL-22R and IL-10R (also known as IL-10R2), while IL-20 and IL-24 bind to and signal through a heterodimeric complex consisting of IL-22R and IL-20R3 (also known as IL-20R2). The IL-22R antibody described in the present invention is in clinical development for atopic dermatitis.

[0058] IL-22 receptor antibodies are described in WO2018011420, in which the HC is SEQ ID NO: 67 and the LC is SEQ ID NO: 68, the VH is SEQ ID NO: 63 and the VL is SEQ ID NO: 64, the HCDR1 is SEQ ID NO: 34 (SYDMN), the HCDR2 is SEQ ID NO: 36 (SIYNDASNTAYSDSVKG) and the HCDR3 is SEQ ID NO: 6 (VGFSGTYYSES), the LCDR1 is SEQ ID NO: 16 (QGGYYAH), the LCDR2 is SEQ ID NO: 47 (GQNNRPS) and the LCDR3 is SEQ ID NO: 54 (QSGSSSSNAV). The SEQ ID NOs refer to the numbers in the above application.

[0059] The following shows the outlines of SEQ ID NOs: 67, 68, 64, and 63. [Table 1]

[0060] According to the present invention, the IL-22 receptor antibodies tested in the following examples are defined as follows: SEQ ID NO: 1: SYDM SEQ ID NO: 2: SIYNDASNTAYSDSVKG Sequence number 3: VGFSGTYYSES Sequence number 4: QGGYYAH Sequence number 5: GQNNRPS Sequence number 6: QSGSSSSNAV Heavy chain (HC) Sequence number 7: QVQLVESGGG LVQPGGSLRL SCAASGFTFS SYDMNWVRQA PGKGLEWVSS IYNDASNTAY SDSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCAKVG FSGTYYSESW GQGTLVTVSS ASTKGPSVFP LAPSSKSTSG GTAALGCLVK DYFPEPVTVS WNSGALTSGV HTFPAVLQSS GLYSLSSVVT VPSSSLGTQT YICNVNHKPS NTKVDKKVEP KSCDKTHTCP PCPAPELLGG PSVFLFPPKP KDTLMISRTP EVTCVVVDVS HEDPEVKFNW YVDGVEVHNA KTKPREEQYQ STYRVVSVLT VLHQDWLNGK EYKCKVSNKA LPAPIEKTIS KAKGQPREPQ VYTLPPSRDE LTKNQVSLTC LVKGFYPSDI AVEWESNGQP ENNYKTTPPV LDSDGSFFLY SKLTVDKSRW QQGNVFSCSV MHEALHNHYT QKSLSLSPGK Light chain (LC) Sequence number 8: SYELTQPSSV SVALGQTARI TCQGGYYAHW YQQKPGQAPV LVIYGQNNRP SGIPERFSGS GAGNTATLTI SRAQAEDEAD YYCQSGSSSS NAVFGGGTKL TVLGQPKAAP SVTLFPPSSE ELQANKATLV CLISDFYPGA VTVAWKADSS PVKAGVETTT PSKQSNNKYA ASSYLSLTPE QWKSHRSYSC QVTHEGSTVE KTVAPTECS Variable heavy chain (VH) Sequence number 9: QVQLVESGGG LVQPGGSLRL SCAASGFTFS SYDMNWVRQA PGKGLEWVSS IYNDASNTAY SDSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCAKVG FSGTYYSESW GQGTLVTVSS Variable light chain: Sequence number 10: SYELTQPSSV SVALGQTARI TCQGGYYAHW YQQKPGQAPV LVIYGQNNRP SGIPERFSGS GAGNTATLTI SRAQAEDEAD YYCQSGSSSS NAVFGGGTKL TVL

[0061] Anti-IL22R, anti-IL-22R, anti-IL22R, anti-IL-22R, etc. used in this specification all refer to antibodies that bind to the IL-22 receptor, i.e., IL-22 receptor antibodies.

[0062] Functional variants (equivalents) of the above IL22R antibodies that have the same epitope-binding specificity as anti-IL-22R and exhibit substantially similar biological activities are also included within the scope of the present invention and are also disclosed in WO2018011420. In some embodiments, the functional variant comprises the same region / residue involved in antigen binding, e.g., the same specificity-determining residues in the CDR or the entire CDR. In other embodiments, the functional variant comprises a VH chain comprising VH CDR1, VH CDR2, and VH CDR3 that are at least 75% (e.g., 80%, 85%, 90%, 95%, or 98%) identical to the corresponding VH CDR of the antibody, and a VL chain comprising VL CDR1, VL CDR2, and VL CDR3 that are at least 75% (e.g., 80%, 85%, 90%, 95%, or 98%) identical to the corresponding VH CDR described above. For example, the functional variant may comprise a VH chain that contains up to 5 (e.g., 1, 2, 3, 4, or 5) amino acid residue mutations in the VH CDR region (the sum of VH CDR1, CDR2, and / or CDR3) compared to the VH CDR, and / or a VL chain that contains up to 5 (e.g., 1, 2, 3, 4, or 5) amino acid residue mutations in the VL CDR region (the sum of VL CDR1, CDR2, and / or CDR3) compared to the above VH CDR. Alternatively, the functional variant comprises a VH chain that is at least 75% (e.g., 80%, 85%, 90%, 95%, or 98%) identical to the VH chain, and a VL chain that is at least 75% (e.g., 80%, 85%, 90%, 95%, or 98%) identical to the VL chain. The amino acid sequence variations can occur only in one or more of the VH and / or VL framework regions. Antibodies having essentially the same characteristics as those described above are expected to be useful in the present invention. Some sequence variations while still maintaining the binding characteristics are mutations covered by the present invention.

[0063] Alternatively or in addition thereto, the amino acid residue mutations can be conservative amino acid residue substitutions. As used herein, "conservative amino acid substitutions" refer to amino acid substitutions that do not change the relative charge or size characteristics of the protein in which the amino acid substitution is made. Variants can be produced according to methods known to those skilled in the art for changing polypeptide sequences, such as those described in references that summarize such methods, such as Molecular Cloning: A Laboratory Manual, J. Sambrook, et al., eds., Second Edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 1989, or Current Protocols in Molecular Biology, F.M. Ausubel, et al., eds., John Wiley & Sons, Inc., New York. Conservative substitutions of amino acids include the following substitutions among amino acids within the following groups: (a) M, I, L, V; (b) F, Y, W; (c) K, R, H; (d) A, G; (e) S, T; (f) Q, N; and (g) E, D.

[0064] The liquid pharmaceutical formulation according to the present invention meets one or more of the following criteria alone or in combination: · A single-use liquid formulation for subcutaneous injection · Storage stability at 5°C for at least 3 years · Stability during use at room temperature · pH related to subcutaneous injection · Osmolality related to subcutaneous injection (close to isotonicity during subcutaneous injection), · Viscosity less than 25 cP at 25°C.

[0065] The liquid pharmaceutical formulation according to the present invention contains an IL-22R antibody, a suitable buffer, an antioxidant, one or more suitable stabilizers, and a nonionic surfactant, and optionally a viscosity reducing agent. The pH of the formulation is about 5.5 to 6.5.

[0066] In the present invention, the stability of a formulation means that the antibody has a measurable tendency to maintain the same monomeric state and / or physical and chemical structure as at the initial time point or a defined reference point in the formulation. In the present invention, a "stable formulation" refers to a formulation in which the physical and / or chemical stability parameters of the antibody are about 80-100% of the initially defined value, which includes retention of at least and about 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, 90%, 89%, 88%, 87%, 86%, 85%, 84%, 83%, 82%, 81% or 80% of the stability parameters. Important stability parameters are related to protein aggregation. Thus, the stability of a protein formulation is evaluated by quantifying, by size exclusion chromatography, the percentage of protein aggregates (percentage of high molecular weight protein, HMWP%), the percentage of monomeric protein, and the percentage of low molecular weight protein (LMWP%) as described in the Examples section. Thermal stability can be evaluated by storing the sample at a high temperature and then performing chromatographic analysis. Thermal stability can also be evaluated by scanning fluorimetry, as described in the Examples section, in which protein denaturation induced by a temperature increase is detected by a change in autofluorescence, or by light scattering, in which the onset of protein aggregation is determined by an increase in scattering intensity, an increase in the estimated hydrodynamic radius or an increase in the estimated molecular weight.

[0067] In the context of the present invention and as described above, "stable" means that the presence of high molecular weight protein is less than 5% after 3 years at 5°C, less than 3% for HMWP when measured at accelerated conditions, for example at 25°C for 6 months, or for further accelerated conditions, for example at 30°C for 4 weeks, the HMWP is less than 3%.

[0068] In the case of a formulation without arginine, more accelerated conditions such as 4 weeks at 40°C can be used, after which the HMWP is less than 4%.

[0069] The usual approach for long-term stability evaluation is data extrapolation. As an example, when storing for 3 months, 6 months, 1 year and possibly 2 years, a linear correlation is shown between time and measurable stability parameters, and using extrapolation, the amount of degraded and intact monomeric protein at later time points can be predicted.

[0070] The analysis of HMWP is performed by size exclusion chromatography. Details are described in Example 3: The protein sample was analyzed by SEC (size exclusion chromatography) using the following. SEC column: Waters BEH 200 SEC, 300 mm × 4.6 mm column. Column temperature: 25 °C. Mobile phase: 100 mM sodium dihydrogen phosphate monohydrate and 200 mM sodium chloride (NaCl). Flow rate: 0.15 mL / min. Detection: 280 nm and 215 nm. SEC integration procedure: HMWP% (percentage of total area of peaks eluting before the monomer peak), LMWP% (percentage of total area of peaks eluting after the monomer peak).

[0071] In one embodiment, the formulation is stable at 5 °C for 2 to 3 years.

[0072] As used herein, the term "viscosity" refers to the magnitude of internal friction in a fluid, and the measured value refers to the resistance to the flow of a liquid formulation when injected through a syringe needle, for example, during administration to a patient. The viscosity of a protein formulation is affected by the protein concentration and the properties of the protein itself, such as sequence and effective surface charge, and by the viscosity of the protein formulation or other components in the formulation, ionic strength, pH and temperature.

[0073] A viscosity reducing agent in the context of the present invention is an additive that reduces the viscosity of the entire formulation as compared to the same formulation without a viscosity reducing additive (when measured under the same environment such as the same pH, the same protein concentration, the same temperature, and the same method). A viscosity reducing agent in the context of the present invention may be an additive that is replaced by another additive (to prevent an undesired increase in osmotic pressure) in order to reduce viscosity. In one embodiment, the viscosity reducing agent reduces the viscosity by at least 10%, in one embodiment at least 20%, in one embodiment at least 30%, in one embodiment at least 40%, in one embodiment at least 50%, in one embodiment at least 60%, and in other embodiments at least 70%.

[0074] As used herein, "viscosity" can be "kinematic viscosity" or "absolute viscosity".

[0075] "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 the same capillary viscometer and allowed to flow by gravity, the more viscous fluid takes longer to flow through the capillary than the less viscous fluid. If one fluid takes 200 seconds to complete the flow and another fluid takes 400 seconds, the second fluid is twice as viscous as the first fluid on the kinematic viscosity scale. "Absolute viscosity", which may also be called dynamic viscosity or simple viscosity, is the product of kinematic viscosity and fluid density: Absolute viscosity = Kinematic viscosity × Density

[0076] Absolute viscosity is expressed in units of centipoise (cP). The SI unit of absolute viscosity is pascal second (Pa*s) or millipascal second (mPa*s), and 1 cP = 1 mPa-s.

[0077] Viscosity is important for the manufacturing processes of active pharmaceutical ingredients and drug products, for example, with regard to ultrafiltration, diafiltration, mixing, and filling into prefilled syringes or self-injectors. Viscosity is also important for patient compliance, for example, with regard to needle gauge and the pressure applied for injection. Viscosity measurements can be performed as described in the examples.

[0078] In one aspect of the invention, the formulation of the antibody is stable and suitable for administration using, for example, a prefilled syringe or a self-injector.

[0079] Osmolality is a measure of water activity, a thermodynamic description of water in a system (relative to pure water), and is controlled by various parameters such as the collective effect of dissolved species (which may also be referred to as solutes, such as proteins and additives). Water activity is sometimes described as a parameter that explains the tendency of water molecules to "escape" from a system compared to their tendency to escape from pure water. The addition of solutes decreases the tendency to escape (osmolality increases). Since osmolality is a measure of water activity, osmolality represents water (not the solute) and is affected by the solute. Osmolality can be measured by the dew point depression of water (vapor pressure osmometry) and the freezing point depression of water. The desired level of osmolality to be achieved can be achieved by the addition of additives such as buffers, salts such as NaCl, arginine-HCl, amino acids (including but not limited to histidine, glycine, arginine, methionine, and proline), saccharides or sugar alcohols (including but not limited to mannitol, trehalose, sucrose). The additives can have both the effect of changing the osmolality and being stabilizers for proteins. Further stabilizers and isotonic agents suitable for adjusting osmolality are described in references such as Handbook of Pharmaceutical Excipients (Fourth Edition, Royal Pharmaceutical Society of Great Britain, Science & Practice Publishers) or Remingtons: The Science and Practice of Pharmacy (Nineteenth Edition, Mack Publishing Company). In the context of the present disclosure, the terms "isosmotic," "isotonic," "substantially isosmotic," and "substantially isotonic" are used interchangeably to refer to formulations having an osmolality in the range of about 270 mOsm / kg to about 450 mOsm / kg. In one embodiment, it is up to about 380 mOsm / kg, or about 270 mOsm / kg to about 370 mOsm / kg, or about 300 mOsm / kg to about 330 mOsm / kg.

[0080] Embodiments of the formulation are isotonic or near isotonic and include those having an osmolality in the range of about 250 to 450 mOsm / kg or 275 to 325 mOsm / kg, and the osmolality includes 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, 359, 360, 361, 362, 363, 364, 365, 366, 367, 368, 369, 370, 371, 372, 373, 374, 375, 376, 377, 378, 379, 380, 381, 382, 383, 384, 385, 386, 387, 388, 389, 390, 391, 392, 393, 394, 395, 396, 397, 398, 399, 400, 401, 402, 403, 404, 405, 406, 407, 408, 409, 410, 411, 412, 413, 414, 415, 516, 417, 418, 419, 420, 421, 422, 423, 424, 425, 526, 427, 428, 429, 430, 431, 432, 433, 434, 435, 436, 437, 438, 439, 440, 441, 442, 443, 444, 445, 446, 447, 448, 449, 450 mOsm / kg.

[0081] Embodiments of the antibody formulation include those described herein that have a low viscosity despite having a high concentration of antibody in solution. Embodiments of the antibody formulation are formulations having a viscosity of less than 25 cP at 25°C. In one embodiment of the formulation, it has a viscosity of 10 - 20 cP at 25°C. Embodiments include formulations having a viscosity of 12 - 15 cP at 25°C. These ranges and values are not limited to the recited numbers and are understood to further include fractional increments.

[0082] Embodiments include a vessel and a pharmaceutical container containing the pharmaceutical formulation described herein. The vessel holds the pharmaceutical formulation of the present invention and can be any suitable vessel known in the art, including vials, bottles, syringes, or any of the various forms well - known in the art for packaging pharmaceutical formulations (including subcutaneous and transdermal delivery devices), but not limited thereto. The syringe may be filled with the pharmaceutical formulation described herein prior to dispensing to the end - user (i.e., a "prefilled syringe"). Embodiments of the present invention include a prefilled syringe containing the pharmaceutical formulation described herein, where the prefilled syringe is in the form of an "auto - injector", and embodiments of the present invention include a prefilled syringe containing the formulation described herein in the form of an "auto - injector". Examples of suitable pen and auto - injector delivery devices include those from companies such as Ypsomed, e.g., "Ypsomate2.25" and "YpsomatePro"; SHL Group, e.g., "Molly"; Owen Mumford, e.g., "Aidaptus" or BD, e.g., "Intevia", but not limited thereto.

[0083] For use of the formulation in in - vivo administration, the formulation must be sterile. The formulation can be sterilized by filtration through a sterile filtration membrane. The therapeutic composition herein is preferably formulated in a single - use prefilled device or auto - injector. The route of administration follows known and acceptable methods, such as single or multiple administrations by subcutaneous administration.

[0084] Buffers are used to control the pH within a range that optimizes the therapeutic effect, particularly when stability is affected by pH. The buffer is preferably present at a concentration in the range of about 5 mM to about 50 mM. Buffers suitable for use in the present invention include both organic and inorganic acids and their salts, as well as amino acids. For example, histidine, citrate, phosphate, succinate, tartrate, fumarate, gluconate, oxalate, lactate, acetate, trimethylamine salts, such as Tris.

[0085] According to experiments conducted with this antibody, the pH should preferably be within 5.5 to 6.5. In one embodiment of the present invention, the stability of the compound is optimal at a pH of about 6.0. In one embodiment of the present invention, the formulation has a pH of about 5.5 to 6.5. In one embodiment, this means any value within the range. Examples are 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4 and 6.5. In one embodiment, the pH is about 6.0.

[0086] In one embodiment of the present invention, the buffer used in the antibody formulation is histidine, and its concentration can be about 5 to 35 mM. In some examples, the concentration of histidine is about 10 to 30 mM, 15 to 25 mM, and in a specific example, the concentration of histidine is about 20 mM.

[0087] Stabilizers are present to regulate or maintain the stability of proteins in the formulation. When stabilizers are used with large biomolecules such as proteins including antibodies, depending on their specific properties, they can interact with charged groups, as well as hydrophilic and hydrophobic groups of amino acid side chains, and hydrophobic patches on the surface of the protein, thereby reducing the possibility of unwanted intermolecular interactions (protein-protein interactions). Stabilizers can also reduce the tendency of protein unfolding / degradation by preferential exclusion from the protein surface. Stabilizers can also increase the chemical stability of the protein.

[0088] The stabilizer can be present in any amount, taking into account the amounts of other components and the osmolarity limit. Stabilizers include polyhydric sugar alcohols, preferably sugar alcohols with three or more hydroxyl groups, such as glycerin, erythritol, arabitol, xylitol, sorbitol, and mannitol. Other typical stabilizers include salts such as NaCl, amino acids such as alanine, glycine, glutamine, asparagine, histidine, methionine, arginine, lysine, ornithine, leucine, 2-phenylalanine, glutamic acid, threonine, proline, etc.; organic saccharides or sugar alcohols such as sucrose, lactose, lactitol, trehalose, stachyose, mannose, sorbose, xylose, ribose, ribitol, myo-inositol, myo-inositol, galactose, galactitol, glycerol, cyclitols (such as inositol), polyethylene glycol, hydrophilic polymers such as polyvinylpyrrolidone; monosaccharides (such as xylose, mannose, fructose, glucose); disaccharides (such as lactose, maltose, sucrose, trehalose); trisaccharides such as raffinose; and polysaccharides such as dextrin or dextran.

[0089] The present invention investigated the optimal use of stabilizers in formulations. In one embodiment of the present invention, the formulations of the present invention are substantially free of NaCl. NaCl is a commonly used stabilizer and isotonic agent, for example, used to adjust osmolarity, increase stability, and decrease viscosity. However, in the present antibody formulations, the presence of NaCl destabilized the antibody.

[0090] The formulations of the present invention showed temperature-dependent stability variations. When tested under high-temperature conditions such as about 50 °C, formulations containing NaCl, arginine, and / or aspartic acid generally contained a larger amount of protein aggregates after heat exposure.

[0091] Also, at lower temperatures such as below 40°C, a larger amount of protein aggregates were present. Generally, when arginine and / or aspartic acid were present, it was considered that a larger amount of antibody aggregates were present after heat exposure. These experiments suggested that arginine and / or aspartic acid should be avoided in the formulation.

[0092] In formulations containing sucrose exposed to high temperatures such as up to 40°C, there were fewer protein aggregates, which indicates the stabilizing effect of sucrose and also shows an improvement in the stabilizing effect of sucrose compared to, for example, mannitol.

[0093] In formulations exposed to high temperatures such as up to 40°C, it was shown that the stabilizing effect was proline > glycine > methionine. However, experiments measuring viscosity showed that formulations containing proline had a higher viscosity.

[0094] It can be seen that when sucrose is present in the formulation, low levels of aggregates are detected across all experiments.

[0095] To meet the requirements regarding viscosity, various experiments examining the viscosity of the formulation were investigated, particularly examining the effect of additives on viscosity and especially the effect of antibody concentration on viscosity. To meet the requirements for viscosity settings, various viscosity reducing agents can be used for testing.

[0096] To accelerate proteolysis and thereby accelerate the progress of formulation development experiments, the stabilization of the formulation is often tested under "accelerated conditions", for example using high temperatures. Temperatures up to 50°C or up to 40°C can be used.

[0097] In such experiments, arginine, which is considered a viscosity reducing agent, was not included in the desired compound by experiment as described above, and it was shown that arginine has a destabilizing effect on the antibody. These data showed that the antibody can be stabilized by sucrose. However, in one experiment, it was observed that the color of the sucrose-containing formulation changed. Therefore, replacing sucrose with trehalose was tested. For various formulations, it was found that replacement of sucrose with trehalose was associated with lower levels of aggregation after storage at 40 °C.

[0098] Although the experimental data clearly showed that arginine enhanced the tendency to aggregate, arginine was included in further experiments.

[0099] After changing the temperature in the stability test, it was found that the destabilizing effect of arginine can be prevented by keeping the temperature below 30 °C. When these conditions were applied, the combination of trehalose and arginine showed the best stabilizing effect in terms of limiting protein aggregation. Surprisingly, arginine had a stabilizing effect when the experiment was conducted at 5 - 25 °C.

[0100] In particular, for formulations with a low antibody concentration, such as about 150 mg / mL, the viscosity reducing agent is about 0 mM, but when the antibody concentration is high, the viscosity reducing agent (especially arginine) is present in the formulation at a concentration of about 75 - 100 mM in addition to the other amino acids present. The concentration of each additive must meet the overall criteria of the osmolality of the formulation.

[0101] When the antibody concentration in the formulation is high, for example 175 - 225 mg / mL, the viscosity reducing agent must be present at an appropriate concentration.

[0102] When the antibody concentration is low, for example 150 - 175 mg / mL, the viscosity of the formulation is low, so the addition of a compound to reduce viscosity may be unnecessary or in small amounts.

[0103] In one embodiment of the present invention, the formulation according to the present invention comprises an IL-22R antibody, a suitable buffer, an antioxidant, one or more suitable stabilizers and a non-ionic surfactant, and optionally a viscosity modifier. In one embodiment, the pH is from about 5.5 to 6.5. In one embodiment, the pH is 6.0. In one embodiment, the buffer is a histidine buffer. In one embodiment, the formulation comprises glycine. In one embodiment, glycine is present up to 80 mM. In other embodiments, the formulation comprises arginine. In one embodiment, arginine is present up to 100 mM. In one embodiment, the antibody is present at 135-175 mg / mL and glycine is present at 80 mM. In one embodiment, the antibody is present at 175-225 mg / mL and glycine is present at 80 mM.

[0104] Nonionic surfactants or detergents are present to prevent surface adsorption, to assist in solubilizing therapeutic proteins, and to protect therapeutic proteins against aggregation induced by agitation and against shear surface stress. Thus, nonionic surfactants can stabilize without causing denaturation of active therapeutic proteins or antibodies (e.g., in contrast to ionic surfactants). Nonionic surfactants are a type of surfactant that do not have a charge on their hydrophilic head group and thus have no net charge. Nonionic surfactants are present in the range of about 0 to about 2 mg / ml, 0.05 mg / ml to about 1.0 mg / ml, preferably about 0.07 mg / ml to about 0.2 mg / ml. Suitable nonionic surfactants include polysorbates (such as 20, 40, 60, 65, 80), polyoxamers (such as 184, 188), Pluronic® polyols (poloxamer 188), Triton®, polyoxyethylene sorbitan monoethers (Tween®-20 (polysorbate 20), Tween®-80 (polysorbate 80), etc.), lauromacrogol 400, polyoxyl 40 stearate, polyoxyethylene hydrogenated castor oils 10, 50 and 60, glycerol monostearate, sucrose fatty acid esters, methylcellulose, and carboxymethylcellulose.

[0105] Suitable nonionic surfactants are polysorbate 20, polysorbate 80, or poloxamer 188. In one embodiment, the nonionic surfactant is present at a concentration of about 0.005 to 1% (w / w). In one embodiment, the concentration of the nonionic surfactant, such as polysorbate 20, can range from 0.005 to 0.5% (w / w). In one embodiment, the nonionic surfactant is polysorbate 20 at a concentration of about 0.01 to 0.08% (w / w). In one embodiment, the nonionic surfactant is polysorbate 20 at a concentration of about 0.01 to 0.03% (w / w). In one embodiment, the concentration of polysorbate 20 is 0.02% (w / w).

[0106] Antibody formulations can be stabilized by the addition of antioxidants. In experiments conducted with the antibodies of the present invention, methionine oxidation was detected at four methionine residues of the anti-IL22R heavy chain (HC), M255 (the most easily oxidized), M34, M83, and M431 (the second most easily oxidized).

[0107] The data are shown in the following table as the total increase in met oxidation for all four positions. The data indicate that the methionine added to the formulation can limit the oxidation of methionine residues. The data support the selection of a methionine concentration in the formulation above 5 mM. The results indicate that antioxidants, such as methionine, can be present at concentrations of 5 to 30 mM. In one embodiment of the present invention, the antioxidant is present at 10 to 25 mM. In one embodiment of the present invention, the antioxidant is present at 15 to 25 mM. In one embodiment of the present invention, the antioxidant is present at 17 to 23 mM. In some embodiments of the present invention, it is about 20 mM. In one embodiment, the methionine concentration in the formulation is >5 mM.

[0108] The anti-IL22R antibody of the present formulation can be present at a concentration of about 135 mg / mL to about 250 mg / mL. In one embodiment, the antibody is present at a concentration of about 150 mg / ml to about 225 mg / mL. In one embodiment, the antibody is present at a concentration of about 140 mg / ml to about 180 mg / mL. In embodiments of the present invention, this means that the antibody is present at 150 mg / mL to 200 mg / mL, or 175 mg / mL to 200 mg / mL, or 150 mg / mL to 175 mg / mL, or about 150 mg / mL, or about 175 mg / mL, or about 200 mg / mL or about 225 mg / mL.

[0109] Dosing regimen and therapeutic applications: The present invention covers a range of antibody concentrations. The formulation meets the set criteria. The formulation is useful for treating diseases responsive to anti-IL-22R treatment.

[0110] To practice the methods described herein, an effective amount of any of the IL-22R antibody formulations described herein can be administered to a subject (e.g., a human) in need of treatment via a suitable route, such as subcutaneous injection or intramuscular injection. The subject to be treated by the methods described herein can be a mammal, more preferably a human. Mammals include, but are not limited to, livestock, sport animals, pets, primates, horses, dogs, cats, mice and rats. A human subject in need of treatment can be a human patient having, at risk of having, or suspected of having a disorder associated with IL-22R. Exemplary IL22R-related disorders include, but are not limited to, inflammatory diseases such as psoriasis, psoriatic arthritis, contact dermatitis and atopic dermatitis. As used herein, "effective amount" refers to the amount of each active substance required to provide a therapeutic effect to a subject, alone or in combination with one or more other active substances. The effective amount will vary depending on parameters of the individual patient including the particular condition being treated, the severity of the condition, age, health status, size, gender and weight, the duration of treatment, the nature of any combination therapy (if any), the particular route of administration, as well as similar factors within the knowledge and expertise of the medical practitioner. The frequency, number and volume of administrations may be determined and adjusted over the course of the treatment and will generally, although not necessarily, be based on the treatment and / or suppression and / or improvement and / or delay of disorders associated with IL-22R. For the purposes of the present disclosure, the appropriate dosage of the IL22R antibody will depend on the particular IL-22R antibody (or composition thereof) being used, the type and severity of the disorder associated with IL-22R, whether the antibody is being administered for prophylactic or therapeutic purposes, previous treatments, the patient's medical history and response to the antibody, as well as the discretion of the attending physician. As used herein, the term "treating" refers to applying or administering a composition comprising one or more active substances to a subject having a disorder associated with IL-22R for the purpose of curing, healing, alleviating, relieving, altering, treating, enhancing, improving or affecting the disorder, symptoms of the disease, or predisposition to the disease.To carry out the methods described herein, any of the anti-IL-22R antibodies can be administered to a subject in need of treatment (e.g., a human patient) by single or multiple administrations via a suitable route, such as subcutaneous injection. The dosage of the anti-IL-22R antibody can range, for example, from about 150 mg / mL in one, two, or three injections of 1 mL, 1.5 mL, 2 mL, 2.5 mL, or 3 mL to about 200 mg / mL in one, two, or three injections of 1 mL, 1.5 mL, 2 mL, 2.5 mL, or 3 mL. Administration of the IL-22R antibody can be a single treatment or repeated administrations at preselected intervals over a period of time in a series of spaced doses.

Example

[0111] All anti-IL22R samples / formulations used to obtain the data of this example were manufactured using Milli-Q water or water for injection (WFI). All pH measurements were performed at room temperature using a calibrated device. The IL22 receptor antibodies used in the following examples are the antibodies defined by SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10.

[0112] Example 1: NaCl cannot be used as an isotonic agent due to its destabilizing effect on anti-IL22R NaCl, a well-known isotonic agent (or osmolarity regulator), cannot be used as an isotonic agent for anti-IL22R due to its destabilizing effect (increased aggregation) on anti-IL22R antibodies.

[0113] Sample preparation: Anti-IL22R was buffer-exchanged into 20 mM histidine at pH 6.5 and the concentration was adjusted to 40 mg / mL. The protein solution was diluted 1:1 with various NaCl solutions (NaCl in Milli-Q water). The final anti-IL22R samples for stability evaluation contained 20 mg / mL anti-IL22R, 10 mM histidine, and NaCl at concentrations of 0 mM, 25 mM, 50 mM, 100 mM, 150 mM, and 300 mM.

[0114] Analysis: Protein samples before and after heat stress were analyzed by SEC (size exclusion chromatography) using the following: SEC column: Waters BEH 200 SEC, 300 mm × 4.6 mm column. Column temperature: 25 °C. Mobile phase: 100 mM sodium dihydrogen phosphate monohydrate and 200 mM sodium chloride (NaCl). Flow rate: 0.15 mL / min. Detection: 280 nm and 215 nm. Run time: 30 minutes. Integration procedure: HMWP% (percent of total area of peaks eluting before the monomer peak), LMWP% (percent of total area of peaks eluting after the monomer peak). Samples were analyzed immediately after production, and after 30 minutes and 1 hour at 60 °C. The SEC data clearly show the correlation between NaCl concentration and the decrease in stability (see the table of SEC integration data below). No chromatographic differences between samples were observed at t = 0, but significant differences were observed after heat exposure.

[0115] Results: Monomer loss during heat exposure is more pronounced in the presence of NaCl. Monomer loss is caused by protein aggregation and is detected as HMWP in the SEC chromatogram. No significant change in LMWP was observed.

Table 2

[0116] Example 2: The thermal stability of anti-IL22R is affected by pH (increases as pH increases) Sample preparation: Anti-IL22R was buffer-exchanged into 20 mM histidine at pH 6.0 and adjusted to a concentration of 2 mg / mL. The protein solution was diluted 1:1 with pH screening solutions at pH values of 3.2, 3.5, 5, 6, 7, 8, 9, and 10. The pH screening solutions contained one of the following two buffers or a mixture of the following two buffers: 1) 100 mM histidine, 100 mM glycylglycine, 100 mM sodium acetate, pH 10, and 2) 100 mM histidine, 100 mM glycylglycine, 100 mM acetic acid, pH 3.2. The final anti-IL22R samples for stability evaluation contained 1 mg / mL anti-IL22R, 60 mM histidine, 50 mM glycylglycine, 50 mM sodium acetate / acetic acid. The sample pH was measured at pH 3.34, pH 3.57, pH 5.09, pH 5.96, pH 6.79, pH 7.64, pH 8.78, pH 9.48.

[0117] Analysis: Samples were exposed to high temperatures from 25 to 85 °C to induce protein aggregation. The onset temperature of aggregation varies between proteins and also depends on the pH and sample composition of each protein. If a protein can withstand high temperatures before starting to aggregate, it is considered thermally stable. The onset temperature of aggregation of anti-IL22R was determined at various pH values. Samples were analyzed by dynamic and static light scattering (DLS and SLS) using 2 × 35 μL per sample in an Aurora 384-well plate ABA2-10100A on a Wyatt Platereader III, software: DYNAMICS version 7.8.2.18. Light scattering data were obtained during heating from 25 to 85 °C. The onset temperature of aggregation (Tagg value) was determined from the light scattering data versus temperature curve. For each sample, it was determined from various data versus temperature traces by the instrument software: 1) Tagg from the estimated hydrodynamic radius versus temperature trace by DLS, 2) Tagg from the estimated molecular weight versus temperature trace by SLS, 3) Tagg from the normalized scattering intensity by SLS. The differences between the Tagg estimates are shown in the result table below.

[0118] Results: The sample pH affects the thermal stability of anti-IL22R. The antibody shows reduced stability at low pH. The highest Tagg values were detected for the anti-IL22R sample at pH 6.8 - 8.8, with the highest value at pH 7.6.

Table 3

[0119] Example 3: The stability of anti-IL22R is affected by pH, and fragmentation in particular increases as the pH increases. Sample preparation: Anti-IL22R was buffer-exchanged into 20 mM histidine at pH 6.5 and adjusted to a concentration of 40 mg / mL. The protein solution was diluted 1:1 with pH screening solutions having pH values of 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, and 8.5. The pH screening solutions were mixtures of the following two buffers: 1) containing 100 mM histidine, 100 mM glycylglycine, 100 mM sodium acetate, and adjusted to pH 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, and 8.5. Samples were analyzed weekly for 3 weeks at 30 °C.

[0120] Analysis: Protein samples were analyzed by SEC (size exclusion chromatography) using the following: SEC column: Waters BEH 200 SEC, 300 mm × 4.6 mm column. Column temperature: 25 °C. Mobile phase: 100 mM sodium dihydrogen phosphate monohydrate and 200 mM sodium chloride (NaCl). Flow rate: 0.15 mL / min. Detection: 280 nm and 215 nm. SEC integration procedure: HMWP% (percentage of total peak area eluting before the monomer peak), LMWP% (percentage of total peak area eluting after the monomer peak). Samples were analyzed immediately after production, and after 6, 7, 14, and 21 days at 30 °C. SEC data show the correlation between pH and mAb degradation (indicating an increase in aggregation / HMWP% and fragmentation / LMWP% (see the following table of SEC integration data).

[0121] Results: The sample pH affects the aggregation, especially fragmentation, of anti-IL22R at 30°C. At 30°C, the antibody is observed to have reduced stability at high pH. The highest HMWP and LMWP values are observed at pH 8.5. Under these experimental conditions, LMWP formation is most prominent, but protein aggregation (formation of HMWP) is considered relatively slow. The stability data obtained indicate that anti-IL22R is most stable at pH 5.5 - 7 with respect to fragmentation. At pH 7.5 - 8.5, anti-IL22R is unstable under the current conditions.

[0122] To confirm the data, anti-IL22R was also analyzed by CE-SDS (instrument: Maurice (ProteinSimple)). Several experimental issues were observed when comparing replicates, such as significant variations in the MW profile (e.g., regarding retention time, peak shape, peak splitting). However, CE-SDS analysis (data not included) confirmed that the fragmentation of anti-IL22R is more prominent at pH > 7.

Table 4

[0123] Comparison of the data from Examples 2 and 3 shows that changing the pH can have opposing effects on various aspects of anti-IL22R stability.

[0124] Example 4: Chemical degradation of anti-IL22R increases with increasing pH Sample preparation: Anti-IL22R was buffer-exchanged into 20 mM histidine at pH 6.5 and adjusted to a concentration of 40 mg / mL. The protein solution was diluted 1:1 with pH screening solutions having pH values of 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, and 8.5. The pH screening solutions were mixtures of the following two buffers: 1) containing 100 mM histidine, 100 mM glycylglycine, 100 mM sodium acetate, and adjusted to pH 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, and 8.5. The samples were analyzed weekly for 2 weeks at 30°C.

[0125] Analysis: The protein sample was analyzed by CIEX (cation exchange chromatography) using the following: CIEX column: MabPac SCX-10 RS 50×2.1 mm, 5 μm. Column temperature: 25 °C. Mobile phase A: Thermo Scientific's diluted CX-1 pH gradient buffer A pH 5.6 (10-fold diluted in Milli-Q water). Mobile phase B: Thermo Scientific's diluted CX-1 pH gradient buffer B pH 10.2 (10-fold diluted in Milli-Q water). Flow rate: 0.4 mL / min Detection: 280 nm and 215 nm. Run time: 23 minutes. Gradient: Initial: Hold 10% B for 2 minutes. Sample separation: 10 - 40% B linear gradient over 13 minutes. Change mobile phase to 90% B and hold for 3 minutes. Return to starting solvent 10% B and hold for 5 minutes. CIEX integration procedure: Acidic peak % (percentage of total area of peaks eluting before the main charge variant), basic peak % (percentage of total area of peaks eluting after the main charge variant). The increase in acidic peak (shown to be related to anti-IL22R deamidation by MS peptide mapping) was quantified as (acidic peak % at t x - acidic peak % at t0). For this mAb, it was observed that the area percentage of the basic peak decreased during the stability test. Therefore, the total chemical change over time was quantified as the sum of the increase in acidic peak and the numerical decrease in basic peak.

[0126] Results: The sample pH affects the chemical degradation of anti-IL22R at 30 °C. At 30 °C, it was observed that the stability of the chemical anti-IL22R gradually decreased with an increase in pH. The chemically quantified increase % in acidic peak area and decrease % in basic peak area are shown in the following table. Increase % in acidic peak area and decrease % in basic peak area at pH 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, and 8.5 (at 30 °C for 6 days and 2 weeks)

Table 5

[0127] Only the increase in the acidic peak (see the table below) suggests that pH 6 provides the highest stability at 30°C over a period of two weeks. % increase in the acidic peak area at pH 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, and 8.5 (for 6 days and 2 weeks at 30°C)

Table 6

[0128] The data shown in Examples 2, 3, and 4 suggest that anti-IL22R is most stable at pH 5.5 - 6.0.

[0129] Example 5: Stabilization and destabilization of anti-IL22R by additives Effect of additives on the tendency of anti-IL22R to denature and aggregate. Fourteen types of additives of various types were tested in this study: sucrose, trehalose, maltose, and lactose (disaccharides, with sucrose and trehalose considered non-reducing sugars and thus most suitable for the development of pharmaceutical liquid protein formulations), histidine, proline, lysine, glycine, glutamic acid (amino acids), mannitol, sorbitol (polyols), sodium phosphate and NaCl (salts), and succinic acid.

[0130] Preparation of samples: Anti-IL22R was buffer-exchanged and concentrated to 56.5 mg / mL in 80 mM histidine at pH 5.98, and the concentration was adjusted to 40.3 mg / mL in 80 mM histidine at pH 5.98. Additive stock solutions were prepared and pH-adjusted to pH 6 ± 0.1. The protein solution was diluted 4-fold with the additive stock solution (75 μL protein solution + 225 μL additive stock solution). The composition of the final protein solution is shown in the table below.

[0131] The samples were analyzed weekly for two weeks at 30°C.

Table 7

[0132] Analysis: The protein sample was analyzed by SEC (size exclusion chromatography) using the following: SEC column: Waters BEH 200 SEC, 300 mm × 4.6 mm column. Column temperature: 25 °C. Mobile phase: 100 mM sodium dihydrogen phosphate monohydrate and 200 mM sodium chloride (NaCl). Flow rate: 0.15 mL / min. Detection: 280 nm. SEC integration procedure: HMWP% (percentage of total peak area of peaks eluting before the monomer peak). The sample was analyzed after manufacture and after exposure to high temperatures (4 weeks at 40 °C and 5 hours, 2 days, and 3 days at 50 °C). The SEC data show how anti-IL22R aggregation (HMWP formation) is affected by various additives.

[0133] The denaturation temperature of anti-IL22R in various samples was analyzed using Uncle (Unchained Labs) by autofluorescence detection during a heating scan. The denaturation temperature (Td) was determined from the inflection point in the fluorescence vs. temperature curve. A higher Td means higher thermal stability.

[0134] Results: The denaturation data (first data column in the table below) show that 100 mM histidine, 100 mM lysine, 100 mM glutamate, 100 mM succinate, 150 mM sodium phosphate, and 150 mM sodium chloride destabilize anti-IL22R by lowering its denaturation temperature and thus increasing the tendency for temperature-induced denaturation. The same additives were observed to increase the aggregate content in the sample when exposed to heat stress at 50 °C for 2 and 3 days and at 40 °C for 4 weeks.

[0135] The SEC data show the stabilizing effect of polyols (mannitol and sorbitol) and disaccharides (sucrose, lactose, maltose, and trehalose) (compared after 4 weeks at 40 °C). Furthermore, 300 mM trehalose and mannitol increase the denaturation temperature of anti-IL22R.

Table 8

[0136] Example 6: Stabilization and destabilization of anti-IL22R by additives Effect of additives on anti-IL22R aggregation at protein concentrations of 65 - 66 mg / mL. During sample preparation, buffer exchange and concentration with an Amicon Ultra-4 mL (30K) spin filter, and centrifugation at 4000 G at room temperature, important observations were made: significant protein loss (anti-IL22R loss) was observed when concentrating a test batch containing 9.49 mg / mL anti-IL22R in 80 mM sodium phosphate, 40 mM sodium chloride, pH 7, but no protein loss was observed when concentrating anti-IL22R in 20 mM histidine at pH 6.5. Thus, the unfavorable effects of sodium chloride and sodium phosphate (e.g., the decrease in thermal stability and the increase in the tendency to aggregate described in the previous examples) also include an increase in surface adsorption during handling and manufacturing processes.

[0137] Samples to be further investigated in this test were prepared by buffer exchange and concentrated into 9 different formulations described in the following table. Subsequently, the protein concentration was adjusted to 65 - 66 mg / mL to equalize all samples with respect to anti-IL22R concentration.

[0138] Analysis: Protein samples were analyzed by SEC (size exclusion chromatography) using the following: SEC column: Waters BEH 200 SEC, 300 mm × 4.6 mm column. Column temperature: 25 °C. Mobile phase: 100 mM sodium dihydrogen phosphate monohydrate and 200 mM sodium chloride (NaCl). Flow rate: 0.15 mL / min. Detection: 280 nm. SEC integration procedure: HMWP% (percentage of the total peak area of peaks eluting before the monomer peak). Samples were analyzed after manufacture and after exposure to high temperatures (4 weeks at 40 °C and 5 hours, 2 days, and 3 days at 50 °C) as shown in the following data table. The SEC data show how anti-IL22R aggregation (HMWP formation) is affected by various additives.

[0139] Results: The SEC data in this experiment showed the destabilizing effects of arginine, aspartic acid and the combination of arginine and aspartic acid, as well as sodium chloride when the anti-IL22R sample was stored at 50 °C (see SEC-HMWP data in the table below). Arginine and NaCl showed a clear destabilizing effect when stored at 40 °C for 4 weeks. In contrast, aspartic acid and the combination of arginine and aspartic acid were not associated with destabilization (increased aggregation) when stored at 40 °C for 4 weeks.

Table 9

[0140] Example 7: Effect of additives on the viscosity of anti-IL22R at a protein concentration of 65 - 66 mg / mL Sample preparation: Buffer exchange and concentration were performed using an Amicon Ultra-4 mL (30K) spin filter and centrifuged at 4000 G at room temperature to obtain 9 different formulations as described in the table below. During sample preparation, glycine and arginine were observed to protect against protein loss. The protein concentration was adjusted to 65 - 66 mg / mL to equalize all samples with respect to anti-IL22R concentration. Viscosity was measured at 22 °C room temperature using a RheoSence microVisc viscometer.

[0141] Results: Generally, the viscosity results showed relatively small differences among the anti-IL22R samples at 65 - 66 mg / mL. The data may indicate that the sucrose, mannitol and proline at the concentrations tested increased the viscosity of the 65 - 66 mg / mL protein samples. The effect of the additive was predicted to increase with increasing anti-IL22R concentration and was confirmed in the following examples.

Table 10

[0142] Example 8: Viscosity as a function of anti-IL22R concentration To examine the effect of anti-IL22R concentration on viscosity, a concentration test was initiated. Anti-IL22R in 60 mg / mL of 20 mM histidine buffer at pH 6.5 was concentrated with an Amicon spin filter (Amicon Ultra-4 mL-MWCo 30K filter), and sampling was performed approximately every 5 - 10 minutes during concentration. The concentration gradually increased over time but became slower at higher concentrations. Six anti-IL22R samples, including the first sample (sample 1), were collected for viscosity measurement. The highest concentration of anti-IL22R obtained in these formulations was 214 mg / mL (sample 6), and further rotation of the sample did not provide a higher protein concentration, suggesting that the solubility of anti-IL22R in 20 mM histidine at pH 6.5 at room temperature is approximately 210 - 220 mg / mL. Viscosity was measured with a RheoSence microVisc viscometer at a shear rate of 1400 s-1 at room temperature of 18 - 22°C. The viscosity data show an exponential increase in viscosity as a function of protein concentration (see the table and figure below).

Table 11

[0143] Example 9: Effect on the Osmolality of Anti-IL22R To investigate the effect of anti-IL22R on osmolality, a concentration test was initiated (osmolality is theoretically a thermodynamic value: a measure of water activity). Since the molar concentration of the protein in the solution is extremely low, the osmolality is expected to be low in a pure protein-aqueous system. Anti-IL22R in 60 mg / mL of 20 mM histidine buffer at pH 6.5 was concentrated using an Amicon spin filter (Amicon Ultra-4 mL-MWCo 30K filter), and sampling was performed approximately every 5 - 10 minutes during concentration. The concentration increased gradually over time but became slower at higher concentrations. Six anti-IL22R samples, including the first sample (sample 1), were collected for osmolality measurement. The highest concentration of anti-IL22R obtained in this formulation was 214 mg / mL (sample 6), and further rotation of the sample did not provide a higher protein concentration, suggesting that the solubility of anti-IL22R in 20 mM histidine at pH 6.5 is approximately 210 - 220 mg / mL. Osmolality was measured by cryoscopic osmometry using an Osmomat 3000 (Gonotec). The osmolality data are shown in the table below. Assuming that 20 mM histidine in these samples contributes an osmolality of 20 mOsm / kg, the data indicate that the osmolality contribution from anti-IL22R at concentrations of 61 - 214 mg / mL (equivalent to 0.4 - 1.5 mM) is approximately 10 - 22 mOsm / kg. Therefore, additives need to be added to obtain an isotonic formulation.

Table 12

[0144] Example 10: Stabilization and destabilization of anti-IL22R by additive combinations The effect of additive combinations on anti-IL22R aggregation was evaluated for 32 different anti-IL22R samples. Sample preparation was carried out using a GE Helthcare MiniTrap DP 10 column and an Amicon Ultra-4mL-MWCO 30K as briefly described below. The target protein concentration was 150 mg / mL. This test ended with two sets of samples, "low-concentration samples" and "high-concentration samples", due to initial challenges in sample preparation at the target protein concentration. "Low-concentration samples" with 46 - 59 mg / mL anti-IL22R were prepared by A). This procedure resulted in significant protein loss. It was found that protein loss could be avoided by changing the production method to procedure B), and as a result, target "high-concentration samples" with 146 - 169 mg / mL anti-IL22R were obtained. Samples with a protein concentration > 156 mg / mL were diluted to 150 mg / mL to obtain a series of samples with smaller protein concentration variations. For all samples, Tween 20 was added after the final step to avoid adsorption of Tween 20 to the column and filter. For all samples, the protein concentration was measured by UV280 (Lunatic (Unchained Labs)).

[0145] A) Anti-IL22R in 60 mg / mL of 20 mM histidine buffer at pH 6.5 was concentrated to 150 mg / mL with an amicon Ultra-4mL-MWCO 30K and subsequently buffer-exchanged into their respective formulations with a MiniTrap DP 10 column (GE Healthcare) (see table below). This procedure resulted in protein loss, and the anti-IL22R concentration became 46 - 59 mg / mL. B) Anti-IL22R in 60 mg / mL of 20 mM histidine buffer at pH 6.5 was buffer-exchanged into their respective formulations with a MiniTrap DP 10 column (GE Healthcare) (see table below). After buffer exchange, the sample (concentration still approximately 60 mg / mL) was concentrated to 150 mg / mL with an amicon Ultra-4mL-MWCO 30K. This procedure yielded anti-IL22R samples of 146 - 169 mg / mL. The samples were diluted to 150 mg / mL with each formulation.

Table 13

[0146] Analysis: Protein samples were analyzed by SEC (size exclusion chromatography) using the following: SEC column: Waters BEH 200 SEC, 300 mm × 4.6 mm column. Column temperature: 25 °C. Mobile phase: 100 mM sodium dihydrogen phosphate monohydrate and 200 mM sodium chloride (NaCl). Flow rate: 0.15 mL / min. Detection: 280 nm. SEC integration procedure: HMWP% (percentage of the total area of peaks eluting before the monomer peak). Samples were analyzed after manufacture, as well as after 5 freeze / thaw cycles and exposure to elevated temperature: 50 °C for 3 days. No clear effect of freeze / thaw stress on HMWP was observed for any of the samples, but the SEC data show how anti-IL22R aggregation (HMWP formation) is affected by the additives. Practically, 0.2 mg / mL Tween 20 corresponds to 0.02% (w / w) Tween 20 (polysorbate 20).

[0147] Results: The SEC data in this experiment show the destabilizing effect of arginine on anti-IL22R (F1 - F7) (see SEC-HMWP data in the table below). In contrast, sucrose is thought to stabilize the antibody (F1, F2, F10 - F15). Thermal-induced anti-IL22R aggregation was found, unsurprisingly, to be protein concentration-dependent (comparison of the third row in the two data tables). Thermal-induced aggregation in anti-IL22R samples containing approximately 50 mg / mL was observed only for some formulations. Based on the data obtained for 150 mg / mL samples, the following conclusions can be drawn. · Formulations containing arginine and / or aspartic acid generally contained a greater amount of protein aggregates after heat exposure. For the heated samples, in the comparison of F2 - F7, there seems to be some correlation between the arginine concentration and the amount of anti - IL22R aggregates (HMWP%). This is consistent with the observations shown in Example 6. · F1 was considered to be more stable than all other arginine - containing formulations, and this was hypothesized to be caused by the stabilizing effect of 180 mM sucrose. · The comparison between F1 and F16 suggests that sucrose is a better stabilizer than mannitol against temperature - induced aggregation. · The comparison between F4 and F5 suggests that glycine is a better stabilizer than methionine against aggregation at high temperatures. · The comparison between F5 and F6 suggests that proline is a better anti - IL22R stabilizer than glycine. · The aggregation of anti - IL22R in F9 - F14 was not as significant compared to all other formulations. These formulations contained various combinations of proline, glycine, methionine, and glycerol. The aggregate content in the F15 sample at high concentration was not determined due to SEC injection failure and lack of material for re - analysis. However, the sample at about 50 mg / mL showed that this sample was also one of the most stable / least aggregated samples. · The aggregation of anti - IL22R in F11 - F14 was not as significant compared to all other formulations, and these formulations contained sucrose.

Table 14

[0148] Example 11: Effects of Additives and pH on Viscosity The effects of additive combinations and anti-IL22R concentrations on viscosity were evaluated for five different anti-IL22R samples. Anti-IL22R in 60 mg / mL of 20 mM histidine buffer at pH 6.5 was buffer-exchanged into each formulation using a MiniTrap DP 10 column (GE Helthcare). After buffer exchange, the samples were concentrated to approximately 150 mg / mL using an amicon Ultra-4 mL-MWCO 30K according to Procedure B described in Example 10. The additive compositions of the investigated formulations F1, F7, F10, F11, and F12 are shown in the prescription table of Example 10 (using the same formulation numbers).

[0149] Viscosity was measured using a RheoSence microVisc viscometer at a shear rate of 1400 s-1 at room temperature of 18 - 22°C. The following table shows the viscosities of samples at various protein concentrations.

Table 15

[0150] All formulations contained 20 mM histidine at pH 6.0 and 0.2 mg / mL Tween 20 (equivalent to 0.02% (w / w) polysorbate 20). The following table outlines the differences between the formulations among F1, F7, F10, F11, and F12.

[0151] The data (the above table) indicate that the lowest viscosity is observed at F7 (this sample contains no disaccharides but contains a combination of four different amino acids: arginine, proline, glycine, and methionine). F1, which contains 180 mM sucrose and 50 mM arginine, has a significantly higher viscosity of 139 mg / mL anti-IL22R compared to F7 (the sucrose-free sample) which has a similar concentration (136 mg / mL). Both of these contain arginine. However, the highest viscosity was seen at F12 which contains proline, methionine, and sucrose. When samples with similar protein concentrations (134 mg / mL and 136 mg / mL respectively) were compared, the viscosity of F10 (which contains 80 mM proline) was slightly higher than that of F11 (which contains no proline but increased sucrose). Thus, this suggests that proline increases the viscosity of anti-IL22R. None of these contain arginine and both contain 80 mM glycine.

Table 16

[0152] Example 12: Effects of Additives and pH on Anti-IL22R Stability The effect of combinations of additives on the biophysical properties of anti-IL22R at approximately 150 mg / mL was evaluated by comparison of 12 different formulations. Twelve placebo formulations (formulations without anti-IL22R and without Tween 20) were manufactured and adjusted to pH 6.0. Anti-IL22R formulations were manufactured using the GRUNT (Unchained Labs) of an automated buffer exchange and concentration system. Approximately 7.5 mL of 109 mg / mL anti-IL22R was used for each formulation. This was buffer exchanged and concentrated to approximately 5 mL of the final formulation. After manufacture of these formulations (before addition of Tween 20), the formulations were sterile filtered. Thereafter, a Tween 20 stock solution was added to reach 0.2 mg / mL in the final formulation and the pH and protein concentration were measured. The composition of the 12 formulations is summarized in the table below.

Table 17

[0153] Analysis: The denaturation temperature of anti-IL22R in various formulations was analyzed by native fluorescence detection during heating scans using Uncle (Unchained Labs). The denaturation temperature (Td) was determined from the inflection point in the fluorescence vs. temperature curve. The viscosity of the anti-IL22R formulations shown in the above table was measured using a RheoSence microVisc viscometer (room temperature 18 - 22 °C, shear rate 1400 s-1). 0.2 mg / ml Tween 20 means 0.02% (w / w) polysorbate 20.

[0154] Results and observations: The viscosities of formulations F1 and F11_i containing 50 mM arginine were lower compared to all other formulations (see the data table below). The highest viscosities were observed for F11_d, F11_f, and F11_h. The comparison of F11_b, F11_d, F11_e, and F11_f suggests that the optimal pH for viscosity is in the range of 5.7 - 6.1. The comparison of F11_b and F11_h suggests that replacing proline with glycine decreases the viscosity of the anti-IL22R formulation.

[0155] The denaturation temperature (T d ) suggests that when the pH range is 5.4 - 6.6, as the pH increases, the thermal stability of anti-IL22R increases. The thermal denaturation test also shows that arginine has a negative effect on the thermal stability of anti-IL22R (the Td of F1 and F11_i is relatively low). This is consistent with the data shown in Examples 6 and 10. Thus, arginine has a positive effect on viscosity (decreasing the viscosity of anti-IL22R) while also having a destabilizing effect. The comparison of T d values for F11_a, F11_b, and F11_c suggests that up to 40 mM methionine has only a negative effect on the thermal stability of anti-IL22R. The comparison of T d values for F1 and F11_i suggests that sucrose is a better stabilizer than glycine.

Table 18

[0156] Further observation: During aseptic filtration, it was observed that the anti-IL22R formulations F11_d and F11_h were extremely difficult to filter. And more importantly, coloring of the samples was observed by visual inspection for most formulations after storage at 40 °C for 8 weeks. Although a detailed root cause investigation was not introduced, MS peptide mapping (data not included) confirmed that sucrose was decomposed and anti-IL22R was glycated. Since the glycosidic bond in trehalose is more stable compared to that in sucrose, it was decided to test replacing sucrose as the disaccharide stabilizer with trehalose as the disaccharide stabilizer. Trehalose has previously been observed to increase the thermal stability of anti-IL22R (Example 5).

[0157] Example 13: Replacement of sucrose with trehalose, a more stable disaccharide The effect of combinations of additives on the biophysical properties of anti-IL22R at approximately 150 mg / mL was evaluated by comparison of 12 different formulations. Twelve placebo formulations (formulations without anti-IL22R and without Tween 20) were manufactured and adjusted to pH 6.0. Anti-IL22R formulations were manufactured using the GRUNT (Unchained Labs) of an automated buffer exchange and concentration system. After manufacture of these formulations (before addition of Tween 20), the formulations were aseptically filtered. Thereafter, a Tween 20 stock solution was added to reach 0.2 mg / mL in the final formulation (same as 0.02% (w / w) polysorbate 20), and the pH and protein concentration were measured. The composition of the 12 formulations is summarized in the following table.

Table 19

[0158] Analysis: The viscosity of the anti-IL22R formulation was measured using a RheoSence microVisc viscometer (room temperature 18 - 22°C, shear rate 1400 s-1). SEC analysis was performed using the following: TSKgel® SuperSW mAb HTP HPLC column. Mobile phase: 100 mM sodium phosphate, 300 mM sodium chloride, pH 6.8. Injection volume: 0.1 μL. UV detection at 280 nm. The undiluted sample was injected and analyzed. SEC integration procedure: HMWP% (percent of total area of peaks eluting before the monomer peak).

Table 20

[0159] Results: The data show that an anti-IL22R formulation of approximately 150 mg / mL containing 50 mM arginine has a viscosity in the range of 7.2 - 7.9 cP at room temperature, while the formulation without arginine has a measured viscosity falling in the range of 9.7 - 11.1 cP.

Table 21

[0160] On the other hand, the SEC data show that the anti-IL22R in the formulation containing 50 mM arginine increased the level of protein aggregates (HMWP%) after 4 and 8 weeks at 40°C (SEC data shown in the table below). Arginine has consistently been shown to destabilize anti-IL22R at high temperatures (Examples 6, 10, and 12). F11_r can be considered a thermostable 150 mg / mL anti-IL22R formulation.

[0161] Interestingly, the long-term data (9 months at 5°C and 9 months at 25°C) revealed that 50 mM arginine can be present without compromising stability when the storage temperature is decreased. The data for the 9-month samples at 5°C actually show that the formulations containing 50 mM arginine (F1, F11_s, and F11_t) have a slightly lower aggregate content compared to the formulations without arginine (F11_r and F11_u).

[0162] Data of samples stored at 25°C for 9 months support this finding (lower HMWP% in the presence of arginine) for formulations containing trehalose as the stabilizing disaccharide, while F1 containing sucrose and arginine has a relatively high level of aggregation.

[0163] Thus, the presented data suggest that the anti-IL22R viscosity is decreased by arginine, the anti-IL22R stability at 40°C is decreased by arginine (due to increased aggregation), surprisingly, the anti-IL22R stability at 5°C is increased by arginine, and the anti-IL22R stability at temperatures above 25°C can be improved by replacing sucrose with trehalose.

Table 22

[0164] Example 14: Viscosity of Heat-Stable Formulations Viscosity curves (viscosity vs. protein concentration) were generated for anti-IL22R formulation F11_r. The formulation was buffer-exchanged and concentrated using a MiniTrap DP 10 column (GE Healthcare) and an amicon Ultra-4mL-MWCO 30K according to procedure B described in Example 11. The additive composition of F11_r, the formulation being examined, is shown in the formulation table of Example 14 (using the same formulation number).

[0165] Analysis: Viscosity at 22°C was measured with a RheoSence microVisc viscometer at room temperature of 18 - 22°C. In this test, the "automatic shear rate" by "Rheosense mvisc" was used for all samples. The actual shear rates used for individual samples (in the range of approximately 50 s-1 to 2400 s-1) are shown in the table below.

Table 23

[0166] Example 15: Methionine The effect of the antioxidant methionine on the stability of anti-IL22R was evaluated by comparison of six formulations containing various concentrations of methionine in the range of 0 - 30 mM. The formulations for this study were also manufactured using the GRUNT of an automated buffer exchange system. Anti-IL22R at 102 mg / mL in histidine buffer at pH 6.0 was buffer-exchanged and concentrated to obtain the formulations outlined below. After buffer exchange and concentration, the formulations were sterile filtered and Tween 20 stock solution was added until it reached 0.2 mg / mL in the final formulation (equivalent to 0.02% (w / w) polysorbate 20). The target pH and protein concentration need to be the same for all samples, with a protein concentration in the range of approximately pH 6 and 150 - 155 mg / mL, and both the pH and protein concentration were measured (values are shown in the last row of the table below). Each formulation was dispensed into prefilled syringes with a fill volume of 1 mL and filling was performed on a LAF bench.

Table 24

[0167] Analysis: Anti-IL22R methionine oxidation was analyzed by reduction of anti-IL22R using peptide mapping after trypsin digestion, followed by reverse-phase chromatography and data-dependent MS / MS detection on a Thermo Q-Exactive hybrid orbitrap mass spectrometer. The MS / MS data was used for confirmation of methionine oxidation positions and the level of methionine oxidation was quantified for the MS data using Protein metrics Byonic / Byologic software. Peptide mapping MS was performed after 2 months at 25 °C and 40 °C and after 7 months at 25 °C. SEC analysis was carried out to quantify the potential effect of methionine on anti-IL22R aggregation (HMWP%). The analysis was performed at the start (t0) and at 1 and 2 months at 5 °C, 25 °C and 40 °C.

[0168] Results: Methionine oxidation was detected at four methionine residues of the anti-IL22R heavy chain (HC), M255 (the most easily oxidized), M34, M83, and M431 (the second most easily oxidized). The data are shown in the following table as the total increase in met oxidation for all four positions. The data clearly show that the methionine added to the formulation can limit the oxidation of methionine residues. The MS data support the selection of a methionine concentration in the formulation above 5 mM. [Table 25]

[0169] SEC data suggest that methionine has a stabilizing effect regarding the prevention of anti-IL22R aggregation. For the formulations exposed to 40 °C, the lowest aggregate content (HMWP%) was observed for the formulation containing 15 mM methionine (1- and 2-month data), while the lowest HMWP levels after storage at 25 °C (2 months) were observed for the formulations containing 20 mM and 30 mM methionine. The HMWP data are shown in the table below. The effect of methionine on fragmentation (LMWP%) was not observed by SEC analysis (however, one sample F11_rc had unexpectedly high LMWP after 1 month at 5 °C, which was considered an outlier and may be due to an error in sample preparation). [Table 26]

[0170] Overall, the data show that 10 - 30 mM methionine is beneficial for the chemical and physical stabilization of anti-IL22R in terms of reducing protein oxidation and aggregation.

[0171] Example 16: Tween 20 (Polysorbate 20) To limit surface adsorption, it is desirable to add Tween 20 to the anti-IL22R formulation (and generally protein formulations). The effect of Tween 20 on the aggregation and viscosity of anti-IL22R was evaluated. Anti-IL22R at 139.5 mg / mL in 20 mM histidine at pH 6.0 was buffer-exchanged into a buffer containing 20 mM histidine, 80 mM glycine, 20 mM methionine, 180 mM trehalose, pH 6.0, using a MiniTrap DP 10 column (GE Healthcare), and concentrated to 150 mg / mL with an amicon Ultra-4mL-MWCO 30K. After buffer exchange and concentration, the formulation was sterile filtered and divided into various aliquots to produce formulations with various Tween 20 concentrations. Tween 20 stock solution was added until it reached 0.2 mg / mL, 0.4 mg / mL, and 0.8 mg / mL (same as 0.02% (w / w), 0.04% (w / w), and 0.08% (w / w) polysorbate 20) in the final formulation, and one formulation was left without Tween 20. The comparative formulations are summarized in the table below. Osmolality, pH, and protein concentration were measured (by cryoscopic osmometry using an Osmomat 3000 (Gonotec), by a standard pH meter, and by a standard UV280 method using lunatic (unchained lab)). SEC analysis and viscosity measurements were performed as described in Example 13.

Table 27

[0172] Results: SEC analysis showed that Tween 20 had no effect on preventing anti-IL22R aggregation. SEC quantified the HMWP% for all samples shown in the table below.

Table 28

[0173] Viscosity was measured for all four formulations, and for F11_w, F11_r, and F11_y, the amount of sample was sufficient to measure viscosity at various concentrations. There was no effect of Tween 20 on viscosity.

Table 29

[0174] Example 17: Effects of Tween 20, Methionine, and pH on a 150 mg / mL Anti-IL22R Formulation In this study, a 150 mg / mL anti-IL22R formulation was prepared and the effects of + / - 0.2 mg / mL Tween 20, + / - 20 mM methionine, pH 6.0 vs. 6.5, and omitted trehalose / glycine concentrations on the long-term anti-IL22R stability at +5 °C and +25 °C and short-term stability at 40 °C were tested. The composition of the formulations is shown in the table below. 102 mg / mL anti-IL22R in 20 mM histidine at pH 6.0 was used as the starting material for the production of various formulations. The protein was diluted to 50 mg / mL with each diafiltration buffer (Tween-free placebo formulation) prior to diafiltration. After diafiltration, a final concentration step to approximately 175 - 200 mg / mL was carried out and the product was replaced with the diafiltration buffer (the target concentration of the final anti-IL22R formulation was approximately 150 mg / mL). After diafiltration and concentration, the formulation was sterile filtered and the Tween 20 stock solution was added until it reached 0.2 mg / mL in the final formulation (equivalent to 0.02% (w / w) polysorbate 20). Osmolality, pH, and protein concentration were measured (as described in Example 16). The formulations were filled into PFS and stored at various temperatures.

Table 30

[0175] Analysis: SEC analysis was performed using the following: Agilent AdvanceBio SEC 200Å, 1.9μm, 4.6mm × 150mm column. Mobile phase: 100 mM sodium phosphate, 300 mM sodium chloride, pH 6.8. Injection volume: 0.1 μL. An undiluted sample was injected and analyzed. UV was detected at 280 nm. SEC integration procedure: HMWP% (percentage of the total area of peaks eluting before the monomer peak). CIEX analysis was performed using the following: MabPac SCX-10 RS 50 × 2.1mm, 5μm. Column temperature: 25°C. Mobile phase A: Thermo Scientific's diluted CX-1 pH gradient buffer A pH 5.6 (10-fold diluted in MilliQ water). Mobile phase B: Thermo Scientific's diluted CX-1 pH gradient buffer B pH 10.2 (10-fold diluted in MilliQ water). Flow rate: 0.4 mL / min. Detection: 280 nm and 215 nm. Run time: 23 minutes. Gradient: Initial: Hold 10% B for 2 minutes. Sample separation: Linear gradient of 10 - 40% B over 13 minutes. Change the mobile phase to 90% B and hold for 3 minutes. Return to the starting solvent 10% B and hold for 5 minutes. CIEX integration procedure: Acidic peak% (percentage of the total area of peaks eluting before the main charge variant), data are shown in the table below. Basic peak% (percentage of the total area of peaks eluting after the main charge variant), data are not shown in the table below as they are not relevant to the noise.

[0176] Results: CIEX analysis shows that the chemical stability is higher at pH 6.0 compared to pH 6.5. This is observed as a greater increase in the acidic peak (compared to the tz value) at most sampling points for formulation F11_ry (CIEX data are shown in the upper table of the following two tables). The increase in the acidic peak area was found to correlate with an increase in deamidation by MS characterization. SEC data indicate that the colloidal stability is also higher at pH 6.0 compared to pH 6.5 (SEC data are shown in the lower table of the following two tables). Comparison of F11_r and F11_ry shows that the HMWP% is consistently higher at pH 6.5, and at some sampling points, the formulation at pH 6.5 was also found to have a greater amount of anti-IL22R fragment, LMWP%.

[0177] Although it could not be used for ranking between various formulations, supplementary information supporting the buffer selection (20 mM histidine): The pH measured at all time points was stable over two years.

Table 31

Table 32

[0178] Example 18: Effect of Arginine on the Viscosity and Stability of Anti-IL22R at Approximately 100 - 200 mg / mL In this test, highly concentrated anti-IL22R formulations with various arginine concentrations were produced and the effect of arginine on viscosity and stability at +5 °C and +25 °C was tested. It was determined to maintain the osmolality at a similar level between formulations (maintaining an osmolality level suitable for sc injection), which means that the concentrations of trehalose and / or glycine decreased with increasing arginine concentration. The composition of the buffer is shown in the following table (note that the F11_s1 placebo formulation in this example has the same composition as F11_s in Example 13). 102 mg / mL anti-IL22R in 20 mM histidine at pH 6.0 was used as the starting material for the production of various formulations. The protein (102 mg / mL for 85 mL) was diluted to 50 mg / mL with each diafiltration buffer (placebo formulation without tween) before diafiltration. After diafiltration, a final concentration step to approximately 200 - 220 mg / mL was carried out and the product was replaced with the diafiltration buffer. After diafiltration and concentration, the formulation was sterile filtered and a tween 20 stock solution was added until it reached 0.2 mg / mL in the final formulation (equivalent to 0.02% (w / w) polysorbate 20). The osmolality (by cryoscopic osmometry using Osmomat 3000 (Gonotec)), pH and protein concentration (by UV absorption using nano drop (Thermo Scientific)) were measured.

Table 33

[0179] Analysis: Viscosity measurements of various concentrations of anti-IL22R in formulations F11_s1, F11_s2, F11_s3, and F11_s4 were performed using a RheoSence microVisc viscometer (at 22 °C and a shear rate of 1400 s-1). By diluting the protein formulations with their respective placebo formulations, an anti-IL22R concentration series for viscosity measurements in the range of approximately 100 - 200 mg / mL was created. The viscosities of high-concentration anti-IL22R in F11_s2, F11_s3, and F11_s4 were further determined using a Dynapro DLS plate reader III (Wyatt) to analyze the effect of temperature on viscosity. The viscosity data are shown in the table below. Stability was evaluated by various methods indicating stability. In this example, CIEX and SEC data are presented to show that there are some differences in stability during 9 months of storage by these analyses. SEC and CIEX analyses were performed as described in Example 17.

[0180] Results: The microVisc-based viscosity data for various concentrations of anti-IL22R in formulations F11_s1, _s2, _s3, and _s4 are shown in the table below. The exponential fit of the viscosity vs. concentration data indicates that anti-IL22R has a similar viscosity profile in F11_s1 and F11_s2, suggesting that in the presence of 50 mM arginine, glycine has no effect on the viscosity reduction of anti-IL22R shown for formulations without arginine (Examples 12 and 13).

[0181] This data also shows that increasing the arginine concentration from 50 mM (F11_s1 and F11_s2) to 80 - 100 mM (F11_s3 and F11_s4) results in a decrease in the viscosity of anti-IL22R.

Table 34

[0182] The effect of arginine on the viscosity of anti-IL22R is even clearer when the present data for the highly concentrated anti-IL22R in F11_s3 and F11_s4 are compared with the viscosity data shown in Example 14 for anti-IL22R in F11_r (arginine-free), where the viscosity increases to about 29 - 48 cP in the concentration range of 195 - 217 mg / mL.

[0183] For highly concentrated formulations at various temperatures, the viscosity was further evaluated. The viscosity increases with decreasing temperature, and the effect of arginine on reducing the viscosity of the anti-IL22R formulation appears more prominently at lower temperatures. From this data, the following effects on viscosity are clear: · Protein concentrations in the range of 200 - 212 mg / mL have a significant effect on viscosity (see data for F11_s3). Therefore, only pairwise comparisons between the 200 mg / mL formulation and the 212 / 213 mg / mL formulation are relevant. · Arginine reduces the viscosity (F11_s2 contains 50 mM arginine, F11_s3 contains 80 mM arginine, and F11_s4 contains 100 mM arginine).

Table 35

[0184] Stability Results: Very similar stability profiles were observed for anti-IL22R in F11_s1, F11_2, F11_3 and F11_4 (see SEC data in the table below). However, after storage at 25°C for 9 months, differences in gastric emptying were revealed by analysis (last / right column in the table below). Comparison of F11_s3 at 212 mg / mL and F11_s4 at 214 mg / mL suggests that F11_s3 is slightly more stable in terms of reduced anti-IL22R aggregation (these samples can be compared as they have similar protein concentrations). The reduction in anti-IL22R aggregation can be caused by a higher concentration of trehalose at 100 mM in F11_s3 compared to 60 mM in F11_s4 (consistent with data from Examples 5 and 13 showing the stabilizing effect of trehalose), and / or by reducing the arginine concentration from 100 to 80 mM. Comparison of F11_s2 at 200 mg / mL and F11_s3 shows that increasing trehalose from 100 mM to 140 mM and simultaneously reducing arginine from 80 mM to 50 mM has a very small effect on the HMWP% after 9 months at 25°C. Thus, the stability of anti-IL22R is considered equivalent in F11_2 and F11_3, but the viscosity data showed the benefit of formulating the mAb in F11_s3 (compared to F11_s2).

[0185] As expected, a slight increase in protein concentration is associated with a slight increase in aggregation. However, for these stable formulations, very small differences were observed when comparing the HMWP values for F11_s3 at 200 mg / mL and 212 mg / mL, and for F11_s4 at 213 mg / mL and 217 - 225 mg / mL. [Table 36] *Some variation was observed in concentration determination

[0186] CIEX data also revealed very similar stability profiles among the formulations, with less increase in acidic charge variants and better maintenance of the main charge variant, indicating that F11_s3 has a lower tendency to be more stable than F11_s1.

Table 37

[0187] In summary, the formulation / concentration data, viscosity data, and stability data guide the formulation balance to achieve a highly concentrated and stable anti-IL22R at approximately 200 - 225 mg / mL.

[0188] Example 19: Effect of Arginine on Viscosity and Injectability In this study, the viscosity and injectability were investigated for various anti-IL22R formulations at various anti-IL22R concentrations. Flexibility regarding injectability and the choice of syringe and needle size is thought to be affected by viscosity. In this study, for various anti-IL22R formulations, injectability tests were performed when injected with a PFS BD NEOPAK 1 mL 1 / 2 inch 27G STW needle or a PFS BD NEOPAK 1 mL 1 / 2 inch 29 TW needle of a prefilled syringe. Measurement of injection force is relevant to the evaluation of the injectability of pharmaceutical formulations. Regulatory guidelines for manual injection propose a maximum injection force of 25 N at 18°C or higher, and as a rule of thumb, subcutaneous injection (regardless of dose) should be performed within 10 seconds.

[0189] The anti-IL22R formulations used in the injectability tests were prepared by manufacturing 102 mg / mL anti-IL22R in 20 mM histidine at pH 6.0 as the starting material. This material was buffer-exchanged (to the formulations shown in the table below), and concentration was carried out using a PD-10 desalting column Sephadex G-25 Medium (GE-Healthcare) and an Amicon Ultra-15 centrifugal filter unit with a MWCO of 50 kDa (Millipore). Four formulations were aseptically filtered through a 0.22 μm filter and subsequently Tween 20 was added.

[0190] Analysis: Viscosity was measured in the same manner as in Example 18. The injection force was measured with 5564 using the following test parameters: an injection volume of 1 mL and an injection rate of 1 mL within 5 seconds. [Table 38]

[0191] The viscosity data shows that the viscosity of F11_r exceeds 20 cP at 181.3 mg / mL anti-IL22R, and that of F11_s2 exceeds 20 cP at 206.8 mg / mL sample, while F11_s3 and F11_s4 do not reach 20 cP even at the highest anti-IL22R concentrations of 205.3 mg / mL and 213.4 mg / mL, respectively. The data shown below represents all the viscosity data for F11_r, F11_s2, F11_s3, and F11_s4 obtained during Examples 18 and 19. [Table 39]

[0192] The injection force data shown in the following table indicates that the injection force depends on the formulation, protein concentration, inner diameter of the needle, and injection time. All experiments were carried out at room temperature of approximately 18 - 20 °C. In this test, it was revealed that most combinations of the formulations, needle thickness, and injection time tested had an injection force of less than 25 N. 1 mL of 200 mg / mL anti-IL22R in F11_s3 and F11_s4 can be easily injected with a 27G STW needle within a preset time of 5 seconds. A force of approximately 18 N (1 mL in 5 seconds) corresponds to the force required to inject 2 mL (400 mg) within 10 seconds. [Table 40]

[0193] Example 20: Stability at 30 °C and 40 °C The anti-IL22R formulation manufactured for the experiments outlined in Example 19 was also used to evaluate anti-IL22R stability at 30 °C and 40 °C (data are shown in this example). Anti-IL22R stability at temperatures above 40 °C has been shown to be impaired by the addition of arginine (see Examples 6, 10, 12, 13), but as shown and described in Example 18, at lower temperatures, the effect of arginine on stability was not as clearly observed. The samples used in this experiment were manufactured according to the procedure described in Example 19.

[0194] Stability at 40 °C. SEC data indicate that the formulation most stable at 40 °C (in terms of having the least amount of protein aggregates and the lowest % HMWP) is F11_r at an anti-IL22R concentration of 151.2 mg / mL. It is most appropriate to compare formulations at similar concentrations. Comparison of formulations containing approximately 175 mg / mL anti-IL22R shows that anti-IL22R is most stable in F11_r after storage at 40 °C for 4 weeks, followed by F11_s2 and F11_s3, and F11_s4 is the least stable at 40 °C. Comparison of F11_s3 and F11_s4 at approximately 200 mg / mL shows that anti-IL22R is most stable in F11_s3.

[0195] Stability at 40 °C. The stability of anti-IL22R at 40 °C is thought to depend on the concentrations of arginine and trehalose. Stability is highest at the lowest arginine concentration (highest trehalose concentration). Thus, F11_r is the formulation that provides the most thermostable anti-IL22R antibody. The viscosity of anti-IL22R in F11_r exceeds 30 cP at room temperature when the protein concentration is 200 mg / mL (see Examples 14 and 19).

[0196] Stability at 30°C. SEC data indicate that at 30°C, there are very small differences between formulations in terms of anti-IL22R stability. At 175 mg / mL anti-IL22R, there is no correlation between stability and arginine concentration (as observed at 40°C), and at 200 mg / mL, F11_s3 is considered to be slightly more stable compared to F11_s4 (consistent with long-term data at 25°C, Example 18). [Table 41]

[0197] All anti-IL22R samples / formulations used to obtain the data in this example were manufactured using Milli-Q water or water for injection (WFI). All pH measurements were performed at room temperature using a calibrated device.

[0198] Example 21: Effect of Tween 20 (Polysorbate 20) on sub-visible particle (SVP) formation in aged anti-IL22R formulations in prefilled syringes The effect of polysorbate 20 on the formation of subvisible particles in a high - protein - concentration anti - IL22R formulation was tested. The analysis was performed on aged formulations as well as aged and stirred formulations to increase the gas - liquid phase stress. The effect of polysorbate 20 was evaluated by micro - flow imaging (MFI, FlowCam). Anti - IL22R was manufactured at 150 ± 15 mg / mL in 20 mM histidine, 80 mM glycine, 20 mM methionine, 180 mM trehalose, pH 6.0. After manufacture, polysorbate 20 was added to a portion of the material to obtain two different formulations: F1 containing 0.02% PS20 (2 mg / mL polysorbate 20), and F2 without surfactant. The formulations were filled into PFS and then exposed to long - term storage at 5°C. The stirring test was performed after storage at 5°C for 3 years. Stirring was carried out at 200 RPM for 3 days (about 72 hours) at room temperature (19 - 23°C) using a 2D shaker plate, both with the syringe lying horizontally and the needle pointing upwards. A non - stirred (static) control was included in the same room.

[0199] Analysis: Analysis was performed using an automated liquid handler and a FlowCam 8100 equipped with a 10x objective lens. The formulations were transferred to 96 - well plates and analyzed without dilution. For each measurement, 150 μL (equivalent to an analysis volume of about 75 μL) was used. Measurements invalidated by background calibration problems or critical fluidity problems were discarded. Particles (e.g., air bubbles) with a circularity of 0.85 or greater were removed during data analysis.

[0200] Results: The following table shows the number of SVPs per milliliter (mean ± standard deviation), grouped into various SVP size groups. N represents the number of independent measurements.

Table 42

[0201] Compared with the F1 formulation containing 0.02% PS20, a large amount of SVPs were observed in the F2 formulation (surfactant-free). Approximately 5 to 30 times more SVPs were observed in F2 compared to F1 (the largest difference in particle number was observed in larger particles). No obvious difference in the number of SVPs was observed between the protein-free F1 and F2 formulations (placebo F1 and placebo F2). Morphological examination of FlowCam SVP images revealed that the particles in F1 did not appear proteinaceous or were only very slightly so. In contrast, the morphology of many SVPs in F2 suggested proteinaceous-type particles. The difference between F1 and F2 observed by FlowCam is thought to be largely independent of agitation. In other words, the particle numbers in F1 and F2 respectively are very similar before and after agitation. The effect of polysorbate 20 on SVP formation in PFS was already evident before agitation (control sample).

[0202] Example 22: Effect of Surfactants on the Formation of Subvisible Particles (SVPs) in an Anti-IL22R Formulation in 2R Vials The effects of non-ionic surfactants (polysorbate 20, polysorbate 80, poloxamer 188) on the formation of subvisible particles in an anti-IL22R formulation with a high protein concentration were tested.

[0203] The influence of surfactants in the range of 0.01 - 0.04% (w / w) (equivalent to 1 - 4 mg / ml) was evaluated by orthogonal techniques (size exclusion chromatography, dynamic light scattering, light obscuration, and microfluidic imaging), and this example shows microfluidic imaging (MFI) data. Anti - IL22R at 150 ± 15 mg / mL was sampled during scale - up production in 20 mM histidine, 80 mM glycine, 20 mM methionine, 180 mM trehalose, pH 6.0. Subsequently, surfactant stock solutions were added to the formulation to obtain various samples containing various surfactants at various concentrations (0.01 - 0.04% (w / w)). 1.6 mL of each formulation was filled into 2R vials. Experimental analysis was performed on newly manufactured formulations (not stability samples as in the previous example), as well as formulations exposed to agitation to increase the gas - liquid phase stress. Agitation was initiated within 24 hours after the addition of the surfactant (placing the 2R vials on a stirring plate). Stirring was carried out at 200 RPM for 3 days (about 72 hours) at room temperature (19 - 23 °C) using a 2D shaker plate. An unstirred control was placed in the same room.

[0204] Analysis: The data shown in this example is based on analysis performed using an automated liquid handler and a FlowCam 8100 equipped with a 10x objective lens. The formulations were transferred to 96 - well plates and analyzed without dilution. For each measurement, 150 μL (equivalent to an analysis volume of about 75 μL) was used. Measurement values invalidated due to background calibration problems or critical fluidity problems were discarded. Particles (e.g., air bubbles) with a circularity of 0.85 or greater were removed during data analysis.

[0205] Results: The following table shows the number of SVPs per milliliter (mean ± standard deviation) divided into various SVP size groups. N represents the number of independent measurements.

Table 43

[0206] Comparison of FlowCam data to a control sample (sample without agitation) shows that the presence of surfactant in the anti-IL22R formulation significantly reduces the number of subvisible particles. The number of particles in freshly manufactured samples (without agitation) is approximately 10 to 20 times higher compared to samples without surfactant. However, the effect of the surfactant is even more pronounced with respect to agitated samples. A general observation is that the effect of the surfactant is crucial in terms of reducing SVP, especially the formation of larger particles during agitation is reduced by the surfactant. For particles >25 μm, the addition of 0.02% (w / w) polysorbate 20 (Tween 20) reduces the number by approximately 300-fold. A nearly identical effect is observed for 0.01% (w / w) polysorbate 80 and 0.02 - 0.04% (w / w) poloxamer 188. Poloxamer 188 was found to be most effective in reducing SVP formation in the test range of 0.01 - 0.04% (w / w) and preventing SVP formation at 0.02% (w / w).

[0207] In summary, the presence of surfactants, namely polysorbate 20, polysorbate 80 and poloxamer 188, has been shown to stabilize the anti-IL22R formulation in a way that inhibits SVP formation.

Claims

1. A liquid pharmaceutical preparation containing IL-22R antibody at a concentration of 150±15 mg / mL to 200±25 mg / mL, as follows: One or more disaccharides, One or more amino acids, cushioning material, Antioxidants, Viscosity reducer, Surfactants, upon request A pharmaceutical preparation further containing [the specified substance], with a pH of 5.5 to 6.5 and an osmolality of 280 to 450 mOsm / kg.

2. It contains IL-22R antibody at concentrations of 150±15 mg / mL to 200±25 mg / mL, as follows: One or more disaccharides in a total concentration of 60–260 mM, One or more amino acids selected from the group consisting of glycine, proline, lysine, glutamic acid, methionine, arginine, aspartic acid, and histidine, in a total concentration of 40–140 mM. Surfactants, upon request The pharmaceutical preparation according to claim 1, further comprising, having a pH of 5.5 to 6.5 and an osmolality of 280 to 450 mOsm / kg.

3. It contains IL-22R antibody at concentrations of 150±15 mg / mL to 200±25 mg / mL, as follows: One or more disaccharides in a total concentration of 60–260 mM, One or more amino acids selected from the group consisting of glycine, proline, lysine, glutamic acid, methionine, arginine, aspartic acid, and histidine, in a total concentration of 40 to 140 mM, wherein the amino acid functions as a stabilizer, antioxidant, viscosity reducer, and buffer. Surfactants, upon request The pharmaceutical preparation according to claim 1, further comprising, having a pH of 5.5 to 6.5 and an osmolality of 280 to 450 mOsm / kg.

4. It contains IL-22R antibody at concentrations of 150±15 mg / mL to 200±25 mg / mL, as follows: One or more disaccharides in a total concentration of 60–260 mM, One or more amino acids selected from the group consisting of glycine, proline, lysine, glutamic acid, methionine, arginine, aspartic acid, and histidine, in a total concentration of 40 to 140 mM, Proline and / or glycine are stabilizers. Methionine is an antioxidant, Arginine is a viscosity-reducing agent. Histidine is a buffer, amino acids, and Surfactants, upon request The pharmaceutical preparation according to claim 1, further comprising, having a pH of 5.5 to 6.5 and an osmolality of 280 to 450 mOsm / kg.

5. The pharmaceutical formulation according to claim 1, wherein the surfactant is present in a concentration of 0.01 to 0.04% (w / w).

6. The pharmaceutical formulation according to claim 5, wherein the surfactant is a nonionic surfactant.

7. The pharmaceutical formulation according to claim 6, wherein the surfactant is selected from the group consisting of polysorbate 20, polysorbate 80, and poloxamer 188.

8. It contains IL-22R antibody at concentrations of 150±15 mg / mL to 200±25 mg / mL, as follows: One or more disaccharides in a total concentration of 60–260 mM, Proline and / or glycine are present at concentrations of 0–80 mM. Methionine is present at concentrations of 5-30 mM. Arginine is present at concentrations of 0-100 mM. Histidine is present at concentrations of 0–30 mM, and Surfactants, upon request The pharmaceutical preparation according to claim 1, further comprising, having a pH of 5.5 to 6.5 and an osmolality of 280 to 450 mOsm / kg.

9. It contains IL-22R antibody at concentrations of 150±15 mg / mL to 200±25 mg / mL, as follows: One or more disaccharides in a total concentration of 60–260 mM, Glycine is present at concentrations of 0-80 mM. Methionine is present at concentrations of 5-30 mM. Arginine is present at concentrations of 0-100 mM. Histidine is present at concentrations of 0–30 mM, and Surfactants, upon request The pharmaceutical preparation according to claim 8, further comprising, having a pH of 5.5 to 6.5 and an osmolality of 280 to 450 mOsm / kg.

10. The pharmaceutical formulation according to claim 8, wherein the surfactant is present in a concentration of 0.01 to 0.04% (w / w).

11. The pharmaceutical formulation according to claim 9, wherein the surfactant is present in a concentration of 0.01 to 0.04% (w / w).

12. The pharmaceutical formulation according to claim 10, wherein the surfactant is selected from the group consisting of polysorbate 20, polysorbate 80, and poloxamer 188.

13. The pharmaceutical formulation according to claim 1, wherein the formulation comprises a histidine buffer.

14. The pharmaceutical formulation according to claim 13, wherein histidine is present at a concentration of 20 mM.

15. The pharmaceutical preparation according to claim 1, wherein the disaccharide is trehalose or sucrose.

16. The pharmaceutical preparation according to claim 15, wherein the disaccharide is trehalose.

17. The pharmaceutical formulation according to claim 1, wherein the surfactant is present in a concentration of 0.01 to 0.02% (w / w).

18. The pharmaceutical formulation according to claim 1, wherein the surfactant is polysorbate 20.

19. The pharmaceutical formulation according to claim 1, wherein the antioxidant methionine is present at a concentration of 20 mM.

20. IL-22R antibody at a concentration of 150±15 mg / mL, and One or more disaccharides at a total concentration of 180–260 mM, One or more amino acids in a total concentration of 40–120 mM, Antioxidants in concentrations of 5-30 mM, Viscosity reducing agent in concentrations of 0 to 100 mM. If desired, surfactants with a total concentration of 0.01 to 0.04% (w / w) 20 mM histidine buffer A pharmaceutical preparation according to claim 1, comprising, having a pH of 5.6 to 6.5 and an osmolality of 280 to 450 mOsm / kg.

21. The pharmaceutical formulation according to claim 20, wherein the surfactant is present in a concentration of 0.01 to 0.03% (w / w).

22. IL-22R antibody at a concentration of 150±15 mg / mL, and Trehalose with a total concentration of 180-260 mM, Glycine in total concentrations of 0-80 mM, Methionine at concentrations of 5-30 mM, Polysorbate 20 with a total concentration of 0.01–0.03% (w / w), 20 mM histidine buffer A pharmaceutical preparation according to claim 21, comprising, having a pH of 5.6 to 6.5 and an osmolality of 280 to 450 mOsm / kg.

23. IL-22R antibody at a concentration of 150±15 mg / mL, and Trehalose with a total concentration of 180-260 mM, Glycine in total concentrations of 0-80 mM, Methionine at concentrations of 5-30 mM, Polysorbate 80 with a total concentration of 0.01–0.03% (w / w), 20 mM histidine buffer A pharmaceutical preparation according to claim 21, comprising, having a pH of 5.6 to 6.5 and an osmolality of 280 to 450 mOsm / kg.

24. IL-22R antibody at a concentration of 150±15 mg / mL, and Trehalose with a total concentration of 180-260 mM, Glycine in total concentrations of 0-80 mM, Methionine at concentrations of 5-30 mM, Poloxamer 188 with a total concentration of 0.01–0.03% (w / w), 20 mM histidine buffer A pharmaceutical preparation according to claim 21, comprising, having a pH of 5.6 to 6.5 and an osmolality of 280 to 450 mOsm / kg.

25. IL-22R antibody at a concentration of 150±15 mg / mL, and Trehalose with a total concentration of approximately 180 mM, Glycine with a total concentration of approximately 80 mM, Methionine at a concentration of approximately 20 mM, Polysorbate 20 with a total concentration of approximately 0.01-0.03% (w / w), Histidine buffer at a concentration of approximately 20 mM A pharmaceutical preparation according to claim 21, comprising and having a pH of 5.6 to 6.

5.

26. It contains IL-22R antibody at a concentration of 200±25 mg / mL, as follows: Trehalose with a total concentration of 60-100 mM, Methionine at concentrations of 5-30 mM, Viscosity reducing agent with a concentration of 60-100 mM, Glycine at concentrations of 0-80 mM, Surfactants, upon request The liquid pharmaceutical preparation according to claim 1, further comprising, having a pH of 5.6 to 6.5 and an osmotic pressure of 280 to 450 mOsm / kg.

27. It contains IL-22R antibody at a concentration of 200±25 mg / mL, as follows: Trehalose with a total concentration of 80-100 mM, Methionine at concentrations of 5-30 mM, Arginine at a concentration of 60-100 mM, Surfactants with a total concentration of 0.01–0.03% (w / w), 20 mM histidine buffer The pharmaceutical preparation according to claim 26, further comprising, having a pH of 5.5 to 6.5 and an osmotic pressure of 280 to 450 mOsm / kg.

28. It contains IL-22R antibody at a concentration of 200±25 mg / mL, as follows: Trehalose with a total concentration of 80-100 mM, Methionine at concentrations of 5-30 mM, Arginine at a concentration of 60-100 mM, 0.00% surfactant, 20 mM histidine buffer The pharmaceutical preparation according to claim 26, further comprising, having a pH of 5.5 to 6.5 and an osmotic pressure of 280 to 450 mOsm / kg.

29. It contains IL-22R antibody at a concentration of 200±25 mg / mL, as follows: Trehalose with a total concentration of approximately 100 mM, Methionine at a concentration of approximately 20 mM, Arginine at a concentration of approximately 80 mM, Polysorbate 20 with a total concentration of approximately 0.02% (w / w), Histidine buffer at a concentration of approximately 20 mM, The pharmaceutical preparation according to claim 26, further comprising and having a pH of 5.5 to 6.

5.

30. It contains IL-22R antibody at a concentration of 200±25 mg / mL, as follows: Trehalose with a total concentration of approximately 100 mM, Methionine at a concentration of approximately 20 mM, Arginine at a concentration of approximately 80 mM, Polysorbate 80 with a total concentration of approximately 0.02% (w / w), Histidine buffer at a concentration of approximately 20 mM, The pharmaceutical preparation according to claim 26, further comprising and having a pH of 5.5 to 6.

5.

31. It contains IL-22R antibody at a concentration of 200±25 mg / mL, as follows: Trehalose with a total concentration of approximately 100 mM, Methionine at a concentration of approximately 20 mM, Arginine at a concentration of approximately 80 mM, Poloxamer 188 with a total concentration of approximately 0.02% (w / w), Histidine buffer at a concentration of approximately 20 mM, The pharmaceutical preparation according to claim 26, further comprising and having a pH of 5.5 to 6.

5.

32. A pharmaceutical preparation according to any one of claims 1 to 31, which is stable for 3 years at 5°C.

33. A pharmaceutical preparation according to any one of claims 1 to 31, which is stable for two years at 5°C.

34. A pharmaceutical preparation according to any one of claims 1 to 31 for use in the treatment of atopic dermatitis.

35. A pharmaceutical formulation according to any one of claims 1 to 31, wherein the IL-22R antibody is defined by the CDR sequences HCDR1: SEQ ID NO: 1, HCDR2: SEQ ID NO: 2, HCDR3: SEQ ID NO: 3, LCDR1: SEQ ID NO: 4, LCDR2: SEQ ID NO: 5, and LCDR3: SEQ ID NO: 6.