Biopharmaceutical composition

Abstract

The present invention provides a composition and a method for improving physical stability of a biopharmaceutical. The present invention involves adding a specific electrolyte (a salt, an acid, or a base, which is in the form of an ion or the like in the present disclosure) and / or cyclodextrin to a dispersive medium for proteins. The present disclosure provides a protein preservative comprising a cyclic oligosaccharide, and at least one electrolyte selected from inorganic salts, organic salts, organic acids, and organic bases, or an ion of the electrolyte.

Description

Biopharmaceutical Compositions

[0001] The present disclosure relates to compositions and methods for improving the physical stability of biopharmaceuticals.

[0002] In developing biopharmaceuticals such as antibody drugs and gene therapy products, it is extremely important to improve the physical stability of proteins, which are the active ingredients.

[0003] In particular, protein aggregates that occur when physical stability is low are impurities that can lead to reduced yields and quality during production, and reduced efficacy and safety during administration. Furthermore, low physical stability can lead to the formation of aggregates during storage in the final product, which can lead to product recall.

[0004] Therefore, in the process of developing biopharmaceutical products, methods are considered for designing the composition of the protein dispersion medium and improving the physical stability of the protein.

[0005] Susumu Uchiyama et al., "Solution Properties of Antibody Drugs," Pharmaceutical Sciences 74 (1), 12-18 (2014)

[0006] The present disclosure is based on the addition of specific electrolytes (organic electrolytes, inorganic electrolytes, or organic or inorganic salts, acids, or bases that constitute them), their ions, etc., and / or cyclic oligosaccharides (e.g., cyclodextrins) to a dispersion medium for proteins.

[0007] Generally, the present disclosure provides the following: (Item 1) A composition for preserving a protein, comprising an electrolyte and / or ions thereof, and a cyclic oligosaccharide or a derivative thereof. (Item 2) A composition comprising a protein, an electrolyte and / or ions thereof, and a cyclic oligosaccharide or a derivative thereof. (Item 3) The composition according to any one of the above items, wherein the composition is for preserving the protein. (Item 4) The composition according to any one of the above items, wherein the electrolyte comprises an organic electrolyte. (Item 5) The composition according to any one of the above items, wherein the electrolyte comprises one or more selected from an inorganic salt, an organic salt, an organic acid, and an organic base. (Item 6) The composition according to any one of the above items, wherein the electrolyte comprises at least one organic acid and at least one organic base. (Item 7) The composition according to any one of the above items, which is in the form of a solution or a solid. (Item 8) The composition according to any one of the above items, wherein the electrolyte is present in an amount of 20 millimoles or more per liter of the total composition. (Item 9) The composition of any one of the above items, wherein the electrolyte is present in an amount of 100 millimoles or more per liter of the total composition. (Item 9A) The composition of any one of the above items, wherein the electrolyte is present in an amount of 20 to 200 millimoles per liter of the total composition. (Item 9B) The composition of any one of the above items, wherein the electrolyte is an organic base and is present in an amount of 20 millimoles or more per liter of the total composition. (Item 9C) The composition of any one of the above items, wherein the electrolyte is an organic base and is present in an amount of 20 to 200 millimoles per liter of the total composition. (Item 9D) The composition of any one of the above items, wherein the electrolyte is present in an amount of 100 to 400 millimoles per liter of the total composition. (Item 10) The composition of any one of the above items, wherein the electrolyte comprises an acid selected from acetic acid, citric acid, tartaric acid, glutamic acid, succinic acid, malic acid, and aspartic acid, or a salt thereof. (Item 11) The composition according to any one of the preceding items, wherein the electrolyte comprises an organic acid selected from acetic acid, succinic acid, and aspartic acid, or a salt thereof.(Item 12) The composition according to any one of the above items, wherein the electrolyte comprises acetic acid or a salt thereof. (Item 13) The composition according to any one of the above items, wherein the electrolyte comprises sodium acetate. (Item 14) The composition according to any one of the above items, wherein the electrolyte comprises at least one organic base selected from arginine, histidine, and tris(hydroxymethyl)aminomethane, or a salt thereof. (Item 15) The composition according to any one of the above items, wherein the organic base is arginine. (Item 16) The composition according to any one of the above items, wherein the electrolyte comprises at least one selected from sodium chloride, potassium chloride, magnesium chloride, calcium chloride, sodium carbonate, sodium phosphate, and potassium phosphate. (Item 17) The composition according to any one of the above items, wherein the electrolyte comprises sodium chloride. (Item 18) The composition according to any one of the above items, wherein the cyclic oligosaccharide or derivative thereof is selected from substituted or unsubstituted (optionally substituted) α-CD, β-CD, and γ-CD. (Item 19) The composition according to any one of the preceding items, wherein the cyclic oligosaccharide or derivative thereof is hydroxypropyl-β-cyclodextrin (HP-β-CD) or sulfobutylether-β-cyclodextrin (SBE-β-CD). (Item 20) The composition according to any one of the preceding items, wherein the cyclic oligosaccharide or derivative thereof is present in an amount of 20 millimoles or more per liter of the total composition. (Item 20A) The composition according to any one of the preceding items, wherein the cyclic oligosaccharide or derivative thereof is present in an amount of 20 millimoles or more and 200 millimoles or less per liter of the total composition. (Item 21) The composition according to any one of the preceding items, wherein the cyclic oligosaccharide or derivative thereof and the electrolyte are contained in equimolar amounts. (Item 22) The composition according to any one of the preceding items, wherein the protein is selected from an antibody, an antigen, a hormone, a cytokine, an enzyme, and a capsid. (Item 23) The particle concentration of the capsid is 1 x 10. １３ The composition according to any one of the preceding items, wherein the capsid particle concentration is 5 x 10 capsid particles / mL or more. １３The composition according to any one of the preceding items, wherein the capsid particles / mL or more. (Item 24) The composition according to any one of the preceding items, wherein the composition is a pharmaceutical composition. (Item 25) A composition for preserving adeno-associated virus (AAV) capsids, comprising an organic electrolyte and / or an ion thereof. (Item 26) A composition for preserving AAV capsids, comprising a cyclic oligosaccharide or a derivative thereof. (Item 27) A composition comprising an AAV capsid and an organic electrolyte or a derivative thereof and / or an ion thereof. (Item 28) A composition comprising an AAV capsid and a cyclic oligosaccharide or a derivative thereof. (Item 29) The composition according to any one of the preceding items, wherein the organic electrolyte comprises at least one organic base and at least one organic acid. (Item 30) The composition according to any one of the preceding items, wherein the organic electrolyte comprises at least one organic base selected from arginine, histidine, and tris(hydroxymethyl)aminomethane. (Item 31) The composition according to any one of the above items, wherein the organic electrolyte comprises at least one organic acid selected from acetic acid, citric acid, tartaric acid, glutamic acid, succinic acid, malic acid, and aspartic acid. (Item 32) The composition according to any one of the above items, wherein the organic electrolyte comprises at least one organic base selected from arginine, histidine, and tris(hydroxymethyl)aminomethane, and at least one organic acid selected from acetic acid, citric acid, tartaric acid, glutamic acid, succinic acid, malic acid, and aspartic acid. (Item 33) The composition according to any one of the above items, wherein the organic electrolyte comprises an organic base that is arginine and an organic acid selected from acetic acid, succinic acid, and aspartic acid. (Item 34) The composition according to any one of the above items, wherein the cyclic oligosaccharide or derivative thereof is selected from substituted or unsubstituted (optionally substituted) α-CD, β-CD, and γ-CD. (Item 35) The composition according to any one of the preceding items, wherein the cyclic oligosaccharide or a derivative thereof is hydroxypropyl-β-cyclodextrin (HP-β-CD) or sulfobutylether-β-cyclodextrin (SBE-β-CD).(Item 36) The composition according to any one of the preceding items, wherein the cyclic oligosaccharide or derivative thereof is present at a concentration of 20 millimoles or more per liter of the total composition. (Item 36A) The composition according to any one of the preceding items, wherein the cyclic oligosaccharide or derivative thereof is present at a concentration of 20 millimoles or more per liter of the total composition and 200 millimoles or less per liter of the total composition. (Item 37) The composition according to any one of the preceding items, wherein the organic electrolyte comprises an organic base and is present at a concentration of 20 millimoles or more per liter of the total composition. (Item 37A) The composition according to any one of the preceding items, wherein the organic electrolyte comprises an organic base and is present at a concentration of 20 millimoles or more per liter of the total composition and 200 millimoles or less per liter of the total composition. (Item 37B) The composition according to any one of the preceding items, wherein the organic electrolyte comprises an organic base and is present at a concentration of 100 millimoles or more per liter of the total composition and 400 millimoles or less per liter of the total composition. (Item 38) The composition according to any one of the preceding items, wherein the organic electrolyte comprises an organic acid and is present at a concentration of 20 millimoles or more per liter of the total composition. (Item 38A) The composition according to any one of the preceding items, wherein the organic electrolyte comprises an organic acid and is present in an amount of 20 to 200 millimoles per liter of the total composition. (Item 38B) The composition according to any one of the preceding items, wherein the organic electrolyte comprises an organic acid and is present in an amount of 100 to 400 millimoles per liter of the total composition. (Item 39) The composition according to any one of the preceding items, further comprising one or more divalent inorganic salts or carbohydrates. (Item 40) The composition according to any one of the preceding items, wherein the divalent inorganic salt is selected from magnesium chloride and calcium chloride. (Item 41) The composition according to any one of the preceding items, wherein the carbohydrate is selected from glucose, sucrose, trehalose, and mannitol. (Item 42) The particle concentration of the AAV capsid is 1 x 10. １３ The composition according to any one of the preceding items, wherein the AAV capsid particle concentration is 5 x 10 capsid particles / mL or more. １３46A) The kit according to any one of the preceding items, wherein the AAV capsid has a capsid density of 10 ... (Item 47) A method for producing the composition according to any one of the above items, the method comprising: 1) providing the protein, and 2) adding the electrolyte and / or the cyclic oligosaccharide or derivative thereof to the protein and mixing. (Item 48) A method for suppressing the formation of protein aggregates using the composition according to any one of the above items, the method comprising: 1) providing the protein, and 2) placing the electrolyte or its ion and / or the cyclic oligosaccharide or derivative thereof under conditions in which it contacts the protein. (Item 49) A method for increasing the concentration of a protein contained in a composition using the composition according to any one of the above items, the method comprising: 1) providing the protein, and 2) placing the electrolyte or its ion and / or the cyclic oligosaccharide or derivative thereof under conditions in which it contacts the protein. (Item A1) Use of an electrolyte and / or its ion and a cyclic oligosaccharide or derivative thereof to preserve a protein. (Item A2) Use of a protein, an electrolyte and / or an ion thereof, and a cyclic oligosaccharide or a derivative thereof. (Item A3) The use according to any one of the above items, wherein the composition is for preserving the protein. (Item A4) The use according to any one of the above items, wherein the electrolyte includes an organic electrolyte.(Item A5) The use according to any one of the above items, wherein the electrolyte comprises one or more selected from inorganic salts, organic salts, organic acids, and organic bases. (Item A6) The use according to any one of the above items, wherein the electrolyte comprises at least one organic acid and at least one organic base. (Item A7) The use according to any one of the above items, wherein the composition according to any one of the above items is a solution or a solid. (Item A8) The use according to any one of the above items, wherein the electrolyte is present in an amount of 20 millimoles or more per liter of the total composition. (Item A9) The use according to any one of the above items, wherein the electrolyte is present in an amount of 100 millimoles or more per liter of the total composition. (Item A9A) The use according to any one of the above items, wherein the electrolyte is present in an amount of 20 millimoles or more and 200 millimoles or less per liter of the total composition. (Item A9B) The use according to any one of the above items, wherein the electrolyte is an organic base and is present in an amount of 20 millimoles or more per liter of the total composition. (Item A9C) The use according to any one of the above items, wherein the electrolyte is an organic base and is present in an amount of 20 to 200 millimoles per liter of the total composition. (Item A9D) The use according to any one of the above items, wherein the electrolyte is present in an amount of 100 to 400 millimoles per liter of the total composition. (Item A10) The use according to any one of the above items, wherein the electrolyte comprises an acid selected from acetic acid, citric acid, tartaric acid, glutamic acid, succinic acid, malic acid, and aspartic acid, or a salt thereof. (Item A11) The use according to any one of the above items, wherein the electrolyte comprises an organic acid selected from acetic acid, succinic acid, and aspartic acid, or a salt thereof. (Item A12) The use according to any one of the above items, wherein the electrolyte comprises acetic acid or a salt thereof. (Item A13) The use according to any one of the above items, wherein the electrolyte comprises sodium acetate. (Item A14) The use according to any one of the above items, wherein the electrolyte comprises at least one organic base selected from arginine, histidine, and tris(hydroxymethyl)aminomethane, or a salt thereof. (Item A15) The use according to any one of the above items, wherein the organic base is arginine.(Item A16) The use according to any one of the above items, wherein the electrolyte comprises at least one selected from sodium chloride, potassium chloride, magnesium chloride, calcium chloride, sodium carbonate, sodium phosphate, and potassium phosphate. (Item A17) The use according to any one of the above items, wherein the electrolyte comprises sodium chloride. (Item A18) The use according to any one of the above items, wherein the cyclic oligosaccharide or derivative thereof is selected from substituted or unsubstituted (optionally substituted) α-CD, β-CD, and γ-CD. (Item A19) The use according to any one of the above items, wherein the cyclic oligosaccharide or derivative thereof is hydroxypropyl-β-cyclodextrin (HP-β-CD) or sulfobutylether-β-cyclodextrin (SBE-β-CD). (Item A20) The use according to any one of the above items, wherein the cyclic oligosaccharide or derivative thereof is present in an amount of 20 mmol or more per liter of the total composition. (Item A20A) The use according to any one of the above items, wherein the cyclic oligosaccharide or derivative thereof is present in an amount of 20 millimoles or more and 200 millimoles or less per liter of the total composition. (Item A21) The use according to any one of the above items, wherein the cyclic oligosaccharide or derivative thereof and the electrolyte are contained in equal molar amounts. (Item A22) The use according to any one of the above items, wherein the protein is selected from an antibody, an antigen, a hormone, a cytokine, an enzyme, and a capsid. (Item A23) The particle concentration of the capsid is 1 x 10 １３ The use according to any one of the preceding items, wherein the capsid particle concentration is 5 x 10 capsid particles / mL or more. １３The use according to any one of the preceding items, wherein the capsid concentration is 1000 capsid particles / mL or more. (Item A24) The use according to any one of the preceding items, wherein the composition according to any one of the preceding items is a pharmaceutical composition. (Item A25) Use of an organic electrolyte and / or ions thereof for preserving adeno-associated virus (AAV) capsids. (Item A26) Use of a cyclic oligosaccharide or a derivative thereof for preserving AAV capsids. (Item A27) Use of an AAV capsid and an organic electrolyte or a derivative thereof and / or ions thereof. (Item A28) Use of an AAV capsid and a cyclic oligosaccharide or a derivative thereof. (Item A29) The use according to any one of the preceding items, wherein the organic electrolyte comprises at least one organic base and at least one organic acid. (Item A30) The use according to any one of the above items, wherein the organic electrolyte comprises at least one organic base selected from arginine, histidine, and tris(hydroxymethyl)aminomethane. (Item A31) The use according to any one of the above items, wherein the organic electrolyte comprises at least one organic acid selected from acetic acid, citric acid, tartaric acid, glutamic acid, succinic acid, malic acid, and aspartic acid. (Item A32) The use according to any one of the above items, wherein the organic electrolyte comprises at least one organic base selected from arginine, histidine, and tris(hydroxymethyl)aminomethane, and at least one organic acid selected from acetic acid, citric acid, tartaric acid, glutamic acid, succinic acid, malic acid, and aspartic acid. (Item A33) The use according to any one of the above items, wherein the organic electrolyte comprises an organic base that is arginine and an organic acid selected from acetic acid, succinic acid, and aspartic acid. (Item A34) The use according to any one of the above items, wherein the cyclic oligosaccharide or derivative thereof is selected from substituted or unsubstituted (optionally substituted) α-CD, β-CD, and γ-CD. (Item A35) The use according to any one of the above items, wherein the cyclic oligosaccharide or derivative thereof is hydroxypropyl-β-cyclodextrin (HP-β-CD) or sulfobutylether-β-cyclodextrin (SBE-β-CD).(Item A36) The use according to any one of the preceding items, wherein the cyclic oligosaccharide or derivative thereof is present at a concentration of 20 millimoles or more per liter of the total composition. (Item A36A) The use according to any one of the preceding items, wherein the cyclic oligosaccharide or derivative thereof is present at a concentration of 20 millimoles or more per liter of the total composition and 200 millimoles or less per liter of the total composition. (Item A37) The use according to any one of the preceding items, wherein the organic electrolyte comprises an organic base and is present at a concentration of 20 millimoles or more per liter of the total composition. (Item A37A) The use according to any one of the preceding items, wherein the organic electrolyte comprises an organic base and is present at a concentration of 20 millimoles or more per liter of the total composition and 200 millimoles or less per liter of the total composition. (Item A37B) The use according to any one of the preceding items, wherein the organic electrolyte comprises an organic base and is present at a concentration of 100 millimoles or more per liter of the total composition and 400 millimoles or less. (Item A38) The use according to any one of the above items, wherein the organic electrolyte comprises an organic acid and is present in an amount of 20 millimoles or more per liter of the total composition. (Item A38A) The use according to any one of the above items, wherein the organic electrolyte comprises an organic acid and is present in an amount of 20 millimoles or more and 200 millimoles or less per liter of the total composition. (Item A38B) The use according to any one of the above items, wherein the organic electrolyte comprises an organic acid and is present in an amount of 100 millimoles or more and 400 millimoles or less per liter of the total composition. (Item A39) The use according to any one of the above items, wherein the composition according to any one of the above items further comprises one or more divalent inorganic salts or carbohydrates. (Item A40) The use according to any one of the above items, wherein the divalent inorganic salt is selected from magnesium chloride and calcium chloride. (Item A41) The use according to any one of the above items, wherein the carbohydrate is selected from glucose, sucrose, trehalose, and mannitol. (Item A42) The particle concentration of the AAV capsid is 1 x 10 １３ The use according to any one of the preceding items, wherein the AAV capsid particle concentration is 5 x 10 capsid particles / mL or more. １３The use according to any one of the above items, wherein the AAV capsid is at least 100 capsid particles / mL. (Item A43) The use according to any one of the above items, wherein the composition according to any one of the above items further comprises one or more surfactants. (Item A44) The use according to any one of the above items, wherein the AAV capsid is an adeno-associated virus vector. (Item A45) Use of a formulation in which the composition according to any one of the above items is filled in a vial or syringe. (Item A46) Use of a kit comprising the composition according to any one of the above items. (Item A46A) Use of the kit according to any one of the above items, wherein the kit contains the components constituting the composition according to any one of the above items in separate compartments. (Item A46B) Use of the kit according to any one of the above items, further comprising instructions describing how to use and / or prepare the composition. (Item AA1) An electrolyte and / or ions thereof and a cyclic oligosaccharide or a derivative thereof for use as a medicine, wherein the electrolyte and / or ions thereof and the cyclic oligosaccharide or a derivative thereof are used to preserve a protein. (Item AA2) A combination of a protein, an electrolyte and / or ions thereof, and a cyclic oligosaccharide or a derivative thereof for use as a medicine. (Item AA3) An electrolyte and / or ions thereof and a cyclic oligosaccharide or a derivative thereof, or a combination thereof, according to any one of the above items, for preserving the protein. (Item AA4) An electrolyte and / or ions thereof and a cyclic oligosaccharide or a derivative thereof, or a combination thereof, according to any one of the above items, wherein the electrolyte comprises an organic electrolyte. (Item AA5) An electrolyte and / or ions thereof and a cyclic oligosaccharide or a derivative thereof, or a combination thereof, according to any one of the above items, wherein the electrolyte comprises one or more selected from inorganic salts, organic salts, organic acids, and organic bases. (Item AA6) The electrolyte and / or ions thereof according to any one of the above items, wherein the electrolyte comprises at least one organic acid and at least one organic base, and a cyclic oligosaccharide or a derivative thereof, or a combination thereof.(Item AA7) The electrolyte and / or ions thereof and cyclic oligosaccharides or derivatives thereof, or combination thereof, according to any one of the preceding items, wherein the electrolyte and / or ions thereof and cyclic oligosaccharides or derivatives thereof are in solution or solid form. (Item AA8) The electrolyte and / or ions thereof and cyclic oligosaccharides or derivatives thereof, or combination thereof, according to any one of the preceding items, wherein the electrolyte is present at 20 millimoles or more per liter of the total composition. (Item AA9) The electrolyte and / or ions thereof and cyclic oligosaccharides or derivatives thereof, or combination thereof, according to any one of the preceding items, wherein the electrolyte is present at 100 millimoles or more per liter of the total composition. (Item AA9A) The electrolyte and / or ions thereof and cyclic oligosaccharides or derivatives thereof, or combination thereof, according to any one of the preceding items, wherein the electrolyte is present at 20 millimoles or more and 200 millimoles or less per liter of the total composition. (Item AA9B) The electrolyte and / or ions thereof according to any one of the preceding items, and a cyclic oligosaccharide or derivative thereof, or a combination thereof, wherein the electrolyte is an organic base and the electrolyte is present in an amount of 20 millimoles or more per liter of the total composition. (Item AA9C) The electrolyte and / or ions thereof according to any one of the preceding items, and a cyclic oligosaccharide or derivative thereof, or a combination thereof, wherein the electrolyte is an organic base and the electrolyte is present in an amount of 20 millimoles or more and 200 millimoles or less per liter of the total composition. (Item AA9D) The electrolyte and / or ions thereof according to any one of the preceding items, and a cyclic oligosaccharide or derivative thereof, or a combination thereof, wherein the electrolyte is present in an amount of 100 millimoles or more and 400 millimoles or less per liter of the total composition. (Item AA10) The electrolyte and / or ions thereof according to any one of the preceding items, wherein the electrolyte comprises an acid selected from acetic acid, citric acid, tartaric acid, glutamic acid, succinic acid, malic acid, and aspartic acid, or a salt thereof, and a cyclic oligosaccharide or a derivative thereof, or a combination thereof. (Item AA11) The electrolyte and / or ions thereof according to any one of the preceding items, wherein the electrolyte comprises an organic acid selected from acetic acid, succinic acid, and aspartic acid, or a salt thereof, and a cyclic oligosaccharide or a derivative thereof, or a combination thereof.(Item AA12) The electrolyte and / or ions thereof according to any one of the preceding items, wherein the electrolyte comprises acetic acid or a salt thereof, and a cyclic oligosaccharide or a derivative thereof, or a combination thereof. (Item AA13) The electrolyte and / or ions thereof according to any one of the preceding items, wherein the electrolyte comprises sodium acetate, and a cyclic oligosaccharide or a derivative thereof, or a combination thereof. (Item AA14) The electrolyte and / or ions thereof according to any one of the preceding items, wherein the electrolyte comprises at least one organic base selected from arginine, histidine, and tris(hydroxymethyl)aminomethane, or a salt thereof, and a cyclic oligosaccharide or a derivative thereof, or a combination thereof. (Item AA15) The electrolyte and / or ions thereof according to any one of the preceding items, wherein the organic base is arginine, and a cyclic oligosaccharide or a derivative thereof, or a combination thereof. (Item AA16) An electrolyte and / or ions thereof according to any one of the preceding items, wherein the electrolyte comprises at least one selected from sodium chloride, potassium chloride, magnesium chloride, calcium chloride, sodium carbonate, sodium phosphate, and potassium phosphate, and a cyclic oligosaccharide or a derivative thereof, or a combination thereof. (Item AA17) An electrolyte and / or ions thereof according to any one of the preceding items, wherein the electrolyte comprises sodium chloride, and a cyclic oligosaccharide or a derivative thereof, or a combination thereof. (Item AA18) An electrolyte and / or ions thereof according to any one of the preceding items, wherein the cyclic oligosaccharide or a derivative thereof is selected from substituted or unsubstituted (optionally substituted) α-CD, β-CD, and γ-CD, and a combination thereof. (Item AA19) The electrolyte and / or ions thereof and a cyclic oligosaccharide or a derivative thereof, or a combination thereof, according to any one of the preceding items, wherein the cyclic oligosaccharide or a derivative thereof is hydroxypropyl-β-cyclodextrin (HP-β-CD) or sulfobutylether-β-cyclodextrin (SBE-β-CD).(Item AA20) The electrolyte and / or ion thereof and cyclic oligosaccharide or derivative thereof, or combination thereof, according to any one of the preceding items, wherein the cyclic oligosaccharide or derivative thereof is present at 20 millimoles or more per liter of the total composition. (Item AA20A) The electrolyte and / or ion thereof and cyclic oligosaccharide or derivative thereof, or combination thereof, according to any one of the preceding items, wherein the cyclic oligosaccharide or derivative thereof is present at 20 millimoles or more and 200 millimoles or less per liter of the total composition. (Item AA21) The electrolyte and / or ion thereof and cyclic oligosaccharide or derivative thereof, or combination thereof, according to any one of the preceding items, wherein the cyclic oligosaccharide or derivative and the electrolyte are contained in equal molar amounts. (Item AA22) The electrolyte and / or ion thereof and cyclic oligosaccharide or derivative thereof, or combination thereof, according to any one of the preceding items, wherein the protein is selected from an antibody, antigen, hormone, cytokine, enzyme, and capsid. (Item AA23) The particle concentration of the capsid is 1 x 10 １３ The electrolyte and / or ions thereof and the cyclic oligosaccharide or derivative thereof, or a combination thereof, according to any one of the above items, wherein the capsid particle concentration is 5 x 10 capsid particles / mL or more. １３(AA24) The electrolyte and / or ion thereof and cyclic oligosaccharide or derivative thereof, or combination thereof, according to any one of the preceding items, wherein the concentration of capsid particles / mL or more is 10 ... (Item AA28) An AAV capsid, an electrolyte and / or an ion thereof, and a cyclic oligosaccharide or a derivative thereof, or a combination thereof. (Item AA29) An electrolyte and / or an ion thereof according to any one of the above items, and a cyclic oligosaccharide or a derivative thereof, or a combination thereof, wherein the organic electrolyte comprises at least one organic base and at least one organic acid. (Item AA30) An electrolyte and / or an ion thereof according to any one of the above items, and a cyclic oligosaccharide or a derivative thereof, or a combination thereof, wherein the organic electrolyte comprises at least one organic base selected from arginine, histidine, and tris(hydroxymethyl)aminomethane. (Item AA31) An electrolyte and / or an ion thereof according to any one of the above items, and a cyclic oligosaccharide or a derivative thereof, or a combination thereof, wherein the organic electrolyte comprises at least one organic acid selected from acetic acid, citric acid, tartaric acid, glutamic acid, succinic acid, malic acid, and aspartic acid.(Item AA32) The electrolyte and / or ions thereof according to any one of the preceding items, wherein the organic electrolyte contains at least one organic base selected from arginine, histidine, and tris(hydroxymethyl)aminomethane, and at least one organic acid selected from acetic acid, citric acid, tartaric acid, glutamic acid, succinic acid, malic acid, and aspartic acid, and a cyclic oligosaccharide or a derivative thereof, or a combination thereof. (Item AA33) The electrolyte and / or ions thereof according to any one of the preceding items, wherein the organic electrolyte contains an organic base that is arginine and an organic acid selected from acetic acid, succinic acid, and aspartic acid, and a cyclic oligosaccharide or a derivative thereof, or a combination thereof. (Item AA34) The electrolyte and / or ions thereof according to any one of the preceding items, wherein the cyclic oligosaccharide or a derivative thereof is selected from substituted or unsubstituted (optionally substituted) α-CD, β-CD, and γ-CD, and a combination thereof. (Item AA35) An electrolyte and / or ions thereof, or a combination of a cyclic oligosaccharide or a derivative thereof, according to any one of the preceding items, wherein the cyclic oligosaccharide or derivative thereof is hydroxypropyl-β-cyclodextrin (HP-β-CD) or sulfobutylether-β-cyclodextrin (SBE-β-CD). (Item AA36) An electrolyte and / or ions thereof, or a combination of a cyclic oligosaccharide or a derivative thereof, according to any one of the preceding items, wherein the cyclic oligosaccharide or derivative thereof is present at a concentration of 20 millimoles or more per liter of the total composition. (Item AA36A) An electrolyte and / or ions thereof, or a combination of a cyclic oligosaccharide or a derivative thereof, according to any one of the preceding items, wherein the cyclic oligosaccharide or derivative thereof is present at a concentration of 20 millimoles or more and 200 millimoles or less per liter of the total composition. (Item AA37) The organic electrolyte comprises an organic base and is present in an amount of 20 millimoles or more per liter of the total composition. An electrolyte and / or an ion thereof and a cyclic oligosaccharide or a derivative thereof, or a combination thereof, according to any one of the above items.(Item AA37A) The electrolyte and / or ions thereof according to any one of the preceding items, and a cyclic oligosaccharide or derivative thereof, or a combination thereof, wherein the organic electrolyte comprises an organic base and is present in an amount of 20 to 200 millimoles per liter of the total composition. (Item AA37B) The electrolyte and / or ions thereof according to any one of the preceding items, and a cyclic oligosaccharide or derivative thereof, or a combination thereof, wherein the organic electrolyte comprises an organic base and is present in an amount of 100 to 400 millimoles per liter of the total composition. (Item AA38) The electrolyte and / or ions thereof according to any one of the preceding items, and a cyclic oligosaccharide or derivative thereof, or a combination thereof, wherein the organic electrolyte comprises an organic acid and is present in an amount of 20 millimoles or more per liter of the total composition. (Item AA38A) The electrolyte and / or ions thereof according to any one of the preceding items, and a cyclic oligosaccharide or derivative thereof, or a combination thereof, wherein the organic electrolyte comprises an organic acid and is present in an amount of 20 to 200 millimoles per liter of the total composition. (Item AA38B) The electrolyte and / or ions thereof according to any one of the above items, and a cyclic oligosaccharide or derivative thereof, or a combination thereof, wherein the organic electrolyte comprises an organic acid and is present in an amount of 100 to 400 millimoles per liter of the total composition. (Item AA39) The electrolyte and / or ions thereof according to any one of the above items, and a cyclic oligosaccharide or derivative thereof, or a combination thereof, further comprising one or more divalent inorganic salts or carbohydrates. (Item AA40) The electrolyte and / or ions thereof according to any one of the above items, and a cyclic oligosaccharide or derivative thereof, or a combination thereof, wherein the divalent inorganic salt is selected from magnesium chloride and calcium chloride. (Item AA41) The electrolyte and / or ions thereof according to any one of the above items, and a cyclic oligosaccharide or derivative thereof, or a combination thereof, wherein the carbohydrate is selected from glucose, sucrose, trehalose, and mannitol. (Item AA42) The particle concentration of the AAV capsid is 1 x 10. １３The electrolyte and / or ions thereof and a cyclic oligosaccharide or a derivative thereof, or a combination thereof, according to any one of the preceding items, wherein the AAV capsid particle concentration is 5 x 10 capsid particles / mL or more. １３(Item AA42) An electrolyte and / or ions thereof and a cyclic oligosaccharide or derivative thereof, or a combination thereof, according to any one of the preceding items, wherein the capsid concentration is 10 ... (Item AA46A) The kit according to any one of the above items, comprising components constituting the electrolyte and / or ions thereof, and the cyclic oligosaccharide or derivative thereof, or combination according to any one of the above items, separated into compartments. (Item AA46B) The kit according to any one of the above items, further comprising instructions describing how to use and / or prepare the electrolyte and / or ions thereof, and the cyclic oligosaccharide or derivative thereof, or combination. (Item AA47) A method for producing the electrolyte and / or ions thereof, and the cyclic oligosaccharide or derivative thereof, or combination according to any one of the above items, comprising the steps of: 1) providing the protein; and 2) adding the electrolyte and / or the cyclic oligosaccharide or derivative thereof to the protein and mixing. (Item AA48) A method for suppressing the formation of aggregates of a protein using an electrolyte and / or ions thereof according to any one of the above items and a cyclic oligosaccharide or a derivative thereof, or a combination thereof, the method comprising: 1) providing the protein; and 2) placing the electrolyte or ions thereof and / or the cyclic oligosaccharide or a derivative thereof under conditions in which the electrolyte or ions thereof and / or the cyclic oligosaccharide or a derivative thereof is brought into contact with the protein.(Item AA49) A method for increasing the concentration of a protein contained in a composition using the electrolyte and / or ions thereof and a cyclic oligosaccharide or derivative thereof, or a combination, according to any one of the above items, the method comprising: 1) providing the protein; and 2) placing the electrolyte or ions thereof and / or cyclic oligosaccharide or derivative thereof under conditions in which the electrolyte or ions thereof and / or cyclic oligosaccharide or derivative thereof contacts the protein. (Item B1) A method for preserving a protein, comprising using an electrolyte and / or ions thereof and a cyclic oligosaccharide or derivative thereof. (Item B2) A method for preserving a protein, comprising using a protein, an electrolyte and / or ions thereof, and a cyclic oligosaccharide or derivative thereof. (Item B3) The method according to any one of the above items, comprising preserving a protein using the composition. (Item B4) The method according to any one of the above items, wherein the electrolyte comprises an organic electrolyte. (Item B5) The method according to any one of the above items, wherein the electrolyte comprises one or more selected from an inorganic salt, an organic salt, an organic acid, and an organic base. (Item B6) The method according to any one of the above items, wherein the electrolyte comprises at least one organic acid and at least one organic base. (Item B7) The method according to any one of the above items, wherein the composition is in the form of a solution or solid. (Item B8) The method according to any one of the above items, wherein the electrolyte is present in an amount of 20 millimoles or more per liter of the total composition. (Item B9) The method according to any one of the above items, wherein the electrolyte is present in an amount of 100 millimoles or more per liter of the total composition. (Item B9A) The method according to any one of the above items, wherein the electrolyte is present in an amount of 20 millimoles or more and 200 millimoles or less per liter of the total composition. (Item B9B) The method according to any one of the above items, wherein the electrolyte is an organic base and is present in an amount of 20 millimoles or more per liter of the total composition. (Item B9C) The method according to any one of the above items, wherein the electrolyte is an organic base and is present in an amount of 20 millimoles or more and 200 millimoles or less per liter of the total composition.(Item B9D) The method of any one of the above items, wherein the electrolyte is present in an amount of 100 millimoles or more and 400 millimoles or less per liter of the total composition. (Item B10) The method of any one of the above items, wherein the electrolyte comprises an acid selected from acetic acid, citric acid, tartaric acid, glutamic acid, succinic acid, malic acid, and aspartic acid, or a salt thereof. (Item B11) The method of any one of the above items, wherein the electrolyte comprises an organic acid selected from acetic acid, succinic acid, and aspartic acid, or a salt thereof. (Item B12) The method of any one of the above items, wherein the electrolyte comprises acetic acid or a salt thereof. (Item B13) The method of any one of the above items, wherein the electrolyte comprises sodium acetate. (Item B14) The method of any one of the above items, wherein the electrolyte comprises at least one organic base selected from arginine, histidine, and tris(hydroxymethyl)aminomethane, or a salt thereof. (Item B15) The method of any one of the above items, wherein the organic base is arginine. (Item B16) The method according to any one of the above items, wherein the electrolyte comprises at least one selected from sodium chloride, potassium chloride, magnesium chloride, calcium chloride, sodium carbonate, sodium phosphate, and potassium phosphate. (Item B17) The method according to any one of the above items, wherein the electrolyte comprises sodium chloride. (Item B18) The method according to any one of the above items, wherein the cyclic oligosaccharide or derivative thereof is selected from substituted or unsubstituted (optionally substituted) α-CD, β-CD, and γ-CD. (Item B19) The method according to any one of the above items, wherein the cyclic oligosaccharide or derivative thereof is hydroxypropyl-β-cyclodextrin (HP-β-CD) or sulfobutylether-β-cyclodextrin (SBE-β-CD). (Item B20) The method according to any one of the above items, wherein the cyclic oligosaccharide or derivative thereof is present in an amount of 20 mmol or more per liter of the total composition. (Item B20A) The method according to any one of the preceding items, wherein the cyclic oligosaccharide or a derivative thereof is present in an amount of 20 mmol to 200 mmol per liter of the total composition.(Item B21) The method according to any one of the above items, wherein the cyclic oligosaccharide or derivative thereof and the electrolyte are contained in equal molar amounts. (Item B22) The method according to any one of the above items, wherein the protein is selected from the group consisting of an antibody, an antigen, a hormone, a cytokine, an enzyme, and a capsid. (Item B23) The particle concentration of the capsid is 1 x 10 １３ The method according to any one of the preceding items, wherein the capsid particle concentration is 5×10 capsid particles / mL or more. １３The method according to any one of the preceding items, wherein the composition according to any one of the preceding items is a pharmaceutical composition. (Item B25) A method for preserving adeno-associated virus (AAV) capsids, comprising using an organic electrolyte and / or ions thereof. (Item B26) A method for preserving AAV capsids, comprising using a cyclic oligosaccharide or a derivative thereof. (Item B27) A method for preserving AAV capsids, comprising using an AAV capsid and an organic electrolyte or a derivative thereof and / or ions thereof. (Item B28) A method for preserving AAV capsids, comprising using an AAV capsid and a cyclic oligosaccharide or a derivative thereof. (Item B29) The method according to any one of the preceding items, wherein the organic electrolyte comprises at least one organic base and at least one organic acid. (Item B30) The method according to any one of the preceding items, wherein the organic electrolyte comprises at least one organic base selected from arginine, histidine, and tris(hydroxymethyl)aminomethane. (Item B31) The method according to any one of the preceding items, wherein the organic electrolyte comprises at least one organic acid selected from acetic acid, citric acid, tartaric acid, glutamic acid, succinic acid, malic acid, and aspartic acid. (Item B32) The method according to any one of the preceding items, wherein the organic electrolyte comprises at least one organic base selected from arginine, histidine, and tris(hydroxymethyl)aminomethane, and at least one organic acid selected from acetic acid, citric acid, tartaric acid, glutamic acid, succinic acid, malic acid, and aspartic acid. (Item B33) The method according to any one of the preceding items, wherein the organic electrolyte comprises an organic base that is arginine and an organic acid selected from acetic acid, succinic acid, and aspartic acid. (Item B34) The method according to any one of the preceding items, wherein the cyclic oligosaccharide or a derivative thereof is selected from substituted or unsubstituted (optionally substituted) α-CD, β-CD, and γ-CD.(Item B35) The method according to any one of the preceding items, wherein the cyclic oligosaccharide or derivative thereof is hydroxypropyl-β-cyclodextrin (HP-β-CD) or sulfobutylether-β-cyclodextrin (SBE-β-CD). (Item B36) The method according to any one of the preceding items, wherein the cyclic oligosaccharide or derivative thereof is present at a concentration of 20 millimoles or more per liter of the total composition. (Item B36A) The method according to any one of the preceding items, wherein the cyclic oligosaccharide or derivative thereof is present at a concentration of 20 millimoles or more per liter of the total composition and 200 millimoles or less per liter of the total composition. (Item B37) The method according to any one of the preceding items, wherein the organic electrolyte comprises an organic base and is present at a concentration of 20 millimoles or more per liter of the total composition. (Item B37A) The method according to any one of the preceding items, wherein the organic electrolyte comprises an organic base and is present at a concentration of 20 millimoles or more per liter of the total composition and 200 millimoles or less per liter of the total composition. (Item B37B) The method according to any one of the preceding items, wherein the organic electrolyte comprises an organic base and is present in an amount of 100 to 400 millimoles per liter of the total composition. (Item B38) The method according to any one of the preceding items, wherein the organic electrolyte comprises an organic acid and is present in an amount of 20 millimoles or more per liter of the total composition. (Item B38A) The method according to any one of the preceding items, wherein the organic electrolyte comprises an organic acid and is present in an amount of 20 to 200 millimoles per liter of the total composition. (Item B38B) The method according to any one of the preceding items, wherein the organic electrolyte comprises an organic acid and is present in an amount of 100 to 400 millimoles per liter of the total composition. (Item B39) The method according to any one of the preceding items, further comprising one or more divalent inorganic salts or carbohydrates. (Item B40) The method according to any one of the preceding items, wherein the divalent inorganic salt is selected from magnesium chloride and calcium chloride. (Item B41) The method according to any one of the above items, wherein the carbohydrate is selected from the group consisting of glucose, sucrose, trehalose, and mannitol. (Item B42) The particle concentration of the AAV capsid is 1 x 10 １３The method according to any one of the preceding items, wherein the AAV capsid particle concentration is 5 x 10 capsid particles / mL or more. １３(Item B46A) A method for producing a kit comprising the composition of any one of the above items. (Item B46B) A method for producing a kit of any one of the above items, further comprising compartmentalized components of the composition of any one of the above items. (Item C46C) A method for producing a kit of any one of the above items, further comprising instructions describing how to use and / or prepare the composition. (Item C1) Use of an electrolyte and / or ions thereof and a cyclic oligosaccharide or a derivative thereof to produce a composition for preserving proteins. (Item C2) Use of these components to produce a composition comprising a protein, an electrolyte and / or an ion thereof, and a cyclic oligosaccharide or a derivative thereof. (Item C3) Use of the composition according to any one of the above items to produce the composition for preserving a protein. (Item C4) Use of the electrolyte comprising an organic electrolyte to produce the composition according to any one of the above items. (Item C5) Use of the electrolyte comprising one or more selected from inorganic salts, organic salts, organic acids, and organic bases to produce the composition according to any one of the above items. (Item C6) Use of the electrolyte comprising at least one organic acid and at least one organic base to produce the composition according to any one of the above items. (Item C7) Use of the composition according to any one of the above items to produce the composition according to any one of the above items in the form of a solution or solid. (Item C8) Use of the electrolyte present in an amount of 20 millimoles or more per liter of the total composition to produce the composition according to any one of the above items. (Item C9) Use of the electrolyte present in an amount of 100 millimoles or more per liter of the total composition for producing the composition according to any one of the preceding items.(Item C9A) Use of the electrolyte, present in an amount of 20 to 200 millimoles per liter of the total composition, for producing the composition of any one of the preceding items. (Item C9B) Use of the electrolyte, wherein the electrolyte is an organic base and is present in an amount of 20 millimoles per liter of the total composition, for producing the composition of any one of the preceding items. (Item C9C) Use of the electrolyte, wherein the electrolyte is an organic base and is present in an amount of 20 to 200 millimoles per liter of the total composition, for producing the composition of any one of the preceding items. (Item C9D) Use of the electrolyte, wherein the electrolyte is present in an amount of 100 to 400 millimoles per liter of the total composition, for producing the composition of any one of the preceding items. (Item C10) Use of the electrolyte comprising an acid selected from acetic acid, citric acid, tartaric acid, glutamic acid, succinic acid, malic acid, and aspartic acid, or a salt thereof, for producing the composition of any one of the preceding items. (Item C11) Use of the electrolyte comprising an organic acid selected from acetic acid, succinic acid, and aspartic acid, or a salt thereof, for producing the composition described in any one of the above items. (Item C12) Use of the electrolyte comprising acetic acid or a salt thereof for producing the composition described in any one of the above items. (Item C13) Use of the electrolyte comprising sodium acetate for producing the composition described in any one of the above items. (Item C14) Use of the electrolyte comprising at least one organic base selected from arginine, histidine, and tris(hydroxymethyl)aminomethane, or a salt thereof, for producing the composition described in any one of the above items. (Item C15) Use of the organic base being arginine for producing the composition described in any one of the above items. (Item C16) Use of the electrolyte comprising at least one selected from sodium chloride, potassium chloride, magnesium chloride, calcium chloride, sodium carbonate, sodium phosphate, and potassium phosphate for producing the composition described in any one of the above items. (Item C17) Use of the electrolyte comprising sodium chloride for producing the composition described in any one of the above items.(Item C18) Use of the cyclic oligosaccharide or its derivative selected from substituted or unsubstituted (optionally substituted) α-CD, β-CD, and γ-CD, for producing the composition described in any one of the above items. (Item C19) Use of the cyclic oligosaccharide or its derivative which is hydroxypropyl-β-cyclodextrin (HP-β-CD) or sulfobutylether-β-cyclodextrin (SBE-β-CD), for producing the composition described in any one of the above items. (Item C20) Use of the cyclic oligosaccharide or its derivative present in an amount of 20 mmol or more per liter of the total composition, for producing the composition described in any one of the above items. (Item C20A) Use of the cyclic oligosaccharide or its derivative present in an amount of 20 mmol or more and 200 mmol or less per liter of the total composition, for producing the composition described in any one of the above items. (Item C21) Use of the cyclic oligosaccharide or its derivative and the electrolyte, wherein the cyclic oligosaccharide or its derivative and the electrolyte are contained in equal molar amounts, for producing the composition described in any one of the above items. (Item C22) Use of a protein selected from an antibody, an antigen, a hormone, a cytokine, an enzyme, and a capsid for producing the composition according to any one of the preceding items. (Item C23) For producing the composition according to any one of the preceding items, a particle concentration of 1 x 10 １３ Use of the capsid particles / mL or more for producing the composition according to any one of the above items. １３Use of the capsid, which has a capsid particle count / mL or more. (Item C24) Use of the ingredients according to any one of the preceding items for producing a pharmaceutical composition. (Item C25) Use of an organic electrolyte and / or ions thereof for producing a composition for preserving adeno-associated virus (AAV) capsids. (Item C26) Use of a cyclic oligosaccharide or a derivative thereof for producing a composition for preserving AAV capsids. (Item C27) Use of these ingredients for producing a composition comprising an AAV capsid and an organic electrolyte or a derivative thereof and / or an ion thereof. (Item C28) Use of these ingredients for producing a composition comprising an AAV capsid and a cyclic oligosaccharide or a derivative thereof. (Item C29) Use of the organic electrolyte comprising at least one organic base and at least one organic acid for producing a composition according to any one of the preceding items. (Item C30) Use of the organic electrolyte comprising at least one organic base selected from arginine, histidine, and tris(hydroxymethyl)aminomethane for producing the composition described in any one of the above items. (Item C31) Use of the organic electrolyte comprising at least one organic acid selected from acetic acid, citric acid, tartaric acid, glutamic acid, succinic acid, malic acid, and aspartic acid for producing the composition described in any one of the above items. (Item C32) Use of the organic electrolyte comprising at least one organic base selected from arginine, histidine, and tris(hydroxymethyl)aminomethane, and at least one organic acid selected from acetic acid, citric acid, tartaric acid, glutamic acid, succinic acid, malic acid, and aspartic acid for producing the composition described in any one of the above items. (Item C33) Use of the organic electrolyte comprising an organic base which is arginine and an organic acid selected from acetic acid, succinic acid, and aspartic acid for producing the composition described in any one of the above items. (Item C34) Use of the cyclic oligosaccharide or a derivative thereof selected from substituted or unsubstituted (optionally substituted) α-CD, β-CD, and γ-CD for producing the composition according to any one of the preceding items.(Item C35) Use of the cyclic oligosaccharide or its derivative, which is hydroxypropyl-β-cyclodextrin (HP-β-CD) or sulfobutylether-β-cyclodextrin (SBE-β-CD), for producing the composition described in any one of the above items. (Item C36) Use of the cyclic oligosaccharide or its derivative, which is present at a concentration of 20 millimoles or more per liter of the total composition, for producing the composition described in any one of the above items. (Item C36A) Use of the cyclic oligosaccharide or its derivative, which is present at a concentration of 20 millimoles or more per liter of the total composition, for producing the composition described in any one of the above items. (Item C37) Use of the organic electrolyte, which comprises an organic base and is present at a concentration of 20 millimoles or more per liter of the total composition, for producing the composition described in any one of the above items. (Item C37A) Use of the organic electrolyte, which comprises an organic base and is present at a concentration of 20 millimoles or more per liter of the total composition, for producing the composition described in any one of the above items. (Item C37B) Use of the organic electrolyte comprising an organic base and present in an amount of 100 to 400 millimoles per liter of the total composition, for producing the composition described in any one of the preceding items. (Item C38) Use of the organic electrolyte comprising an organic acid and present in an amount of 20 millimoles or more per liter of the total composition, for producing the composition described in any one of the preceding items. (Item C38A) Use of the organic electrolyte comprising an organic acid and present in an amount of 20 to 200 millimoles per liter of the total composition, for producing the composition described in any one of the preceding items. (Item C38B) Use of the organic electrolyte comprising an organic acid and present in an amount of 100 to 400 millimoles per liter of the total composition, for producing the composition described in any one of the preceding items. (Item C39) Use of these components to produce the composition described in any one of the preceding items further comprising one or more divalent inorganic salts or carbohydrates. (Item C40) Use of the divalent inorganic salt selected from magnesium chloride and calcium chloride, for producing the composition described in any one of the preceding items.(Item C41) Use of the carbohydrate selected from glucose, sucrose, trehalose, and mannitol for producing the composition according to any one of the above items. (Item C42) For producing the composition according to any one of the above items, a particle concentration of 1 x 10 is used. １３ Use of the AAV capsid having a particle concentration of 5 x 10 capsid particles / mL or more for producing the composition according to any one of the above items. １３(Item C46) Use of the composition of any one of the above items for producing a kit comprising the composition of any one of the above items. (Item C46A) Use of the composition of any one of the above items for producing the kit containing the components of the composition of any one of the above items in separate compartments. (Item C46B) Use of the composition of any one of the above items for producing the kit of any one of the above items, further comprising instructions describing how to use and / or prepare the composition. (Item C47) Use of the composition in producing the composition according to any one of the above items, wherein the composition is produced by a method comprising: 1) providing the protein, and 2) adding the electrolyte and / or the cyclic oligosaccharide or derivative thereof to the protein and mixing. (Item C48) Use of the composition according to any one of the above items in a method for suppressing the formation of protein aggregates, wherein the method comprises: 1) providing the protein, and 2) placing the electrolyte or an ion thereof and / or the cyclic oligosaccharide or a derivative thereof under conditions in which the electrolyte or an ion thereof and / or the cyclic oligosaccharide or a derivative thereof is brought into contact with the protein. (Item C49) Use of the composition according to any one of the above items in a method for increasing the concentration of a protein contained in a composition, wherein the method comprises: 1) providing the protein, and 2) placing the electrolyte or an ion thereof and / or the cyclic oligosaccharide or a derivative thereof under conditions in which the electrolyte or an ion thereof and / or the cyclic oligosaccharide or a derivative thereof is brought into contact with the protein.

[0008] It is contemplated that the present disclosure may provide one or more of the above-described features in combinations other than those explicitly stated. Still further embodiments and advantages of the present disclosure will be recognized by those skilled in the art upon reading and understanding the following detailed description, if necessary.

[0009] Provided are additives and formulation compositions that improve the stability of biopharmaceuticals such as antibody drugs and gene therapy products.

[0010] FIG. 1-1 shows the results of Example 1. As a representative example, it shows the temperature dependence of scattered light intensity in a sample solution of 1.5 mg / mL IgG, 5 mM Cit / NaOH, pH 6.0. The protein aggregation onset temperature (T ａｇｇ ) was calculated and used as an index of protein aggregation. Figure 1-2 shows the results of Example 1. As a representative example, １３ λ in a sample solution of vg / mL AAV6-CMV-EGFP, 10 mM His / HCl, 190 mM NaCl, 0.33% (w / v) PX188, pH 6.0 ＢＣＭ The temperature dependence of λ is shown. ＢＣＭ The protein denaturation temperature (T ｍ ) was calculated using UNcleAnalysis software (V5.03) and used as an index of the stability of the protein's three-dimensional structure, which is known to have a significant effect on protein aggregation. Figure 2-1 shows the results of Example 2. The temperature dependence of the scattered light intensity of AAV6 samples under four conditions, namely, free, 100 mM NaCl, 100 mM HP-β-CD, and 100 mM NaCl + 100 mM HP-β-CD, is compared. Figure 2-2 shows the results of Example 2. The temperature dependence of the scattered light intensity of AAV6 samples under four conditions, namely, free, 100 mM KCl, 100 mM HP-β-CD, and 100 mM KCl + 100 mM HP-β-CD, is compared. Figure 2-3 shows the results of Example 2. The temperature dependence of the scattered light intensity of AAV6 samples under four conditions, namely, free, 100 mM KCl, 100 mM HP-β-CD, and 100 mM KCl + 100 mM HP-β-CD, is compared. ２ , 100 mM HP-β-CD, and 100 mM MgCl ２Figure 2-4 shows the results of Example 2. Figure 2-5 shows the results of Example 1. Figure 2-6 shows the results of Example 1. Figure 2-7 shows the results of Example 2. Figure 2-8 shows the results of Example 3. Figure 2-9 shows the results of Example 4. Figure 2-10 shows the results of Example 4. Figure 2-11 shows the results of Example 4. Figure 2-12 shows the results of Example 4. Figure 2-13 shows the results of Example 4. Figure 2-14 shows the results of Example 4. Figure 2-15 shows the results of Example 4. Figure 2-16 shows the results of Example 4. Figure 2-17 shows the results of Example 4. Figure 2-18 shows the results of Example 4. Figure 2-19 shows the results of Example 5. Figure 2-20 shows the results of Example 5. The temperature dependence of scattered light intensity of AAV6 samples under four conditions, namely, free, 100 mM Na-Ace, 100 mM HP-β-CD, and 100 mM Na-Ace + 100 mM HP-β-CD, is shown. Figure 3-3 shows the results of Example 3. The temperature dependence of scattered light intensity of AAV6 samples under four conditions, namely, free, 100 mM Na-Asp, 100 mM HP-β-CD, and 100 mM Na-Asp + 100 mM HP-β-CD, is shown. Figure 3-4 shows the results of Example 3. The temperature dependence of scattered light intensity of AAV6 samples under four conditions, namely, free, 100 mM Na-Suc, 100 mM HP-β-CD, and 100 mM Na-Suc + 100 mM HP-β-CD, is shown. Figure 3-5 shows the results of Example 3. The temperature dependence of scattered light intensity of AAV6 samples under four conditions: free, 100 mM Na-Mal, 100 mM HP-β-CD, and 100 mM Na-Mal + 100 mM HP-β-CD is compared. Figure 3-6 shows the results of Example 3. The temperature dependence of scattered light intensity of AAV6 samples under four conditions: free, 100 mM Na-Tar, 100 mM HP-β-CD, and 100 mM Na-Tar + 100 mM HP-β-CD is compared. Figure 3-7 shows the results of Example 3. The temperature dependence of scattered light intensity of AAV6 samples under four conditions: free, 100 mM Na-Cit, 100 mM HP-β-CD, and 100 mM Na-Cit + 100 mM HP-β-CD is compared. FIG. 4-1 shows the results of Example 4.Figure 4-2 shows the results of Example 4. Figure 4-3 shows the results of Example 4. The temperature dependence of scattered light intensity of AAV6 samples under four conditions: free, 100 mM Arg-HCl, 100 mM HP-β-CD, and 100 mM Arg-HCl + 100 mM HP-β-CD is compared. Figure 4-4 shows the results of Example 4. The temperature dependence of scattered light intensity of AAV6 samples under four conditions: free, 100 mM Arg-HCl, 100 mM HP-β-CD, and 100 mM Arg-Ace + 100 mM HP-β-CD is compared. The temperature dependence of scattered light intensity of AAV6 samples under four conditions: free, 100 mM Arg-Suc, 100 mM HP-β-CD, and 100 mM Arg-Suc + 100 mM HP-β-CD is shown. Figure 4-4 shows the results of Example 4. The temperature dependence of scattered light intensity of AAV6 samples under four conditions: free, 100 mM Arg-Asp, 100 mM HP-β-CD, and 100 mM Arg-Asp + 100 mM HP-β-CD is shown. Figure 5-1 shows the results of Example 5. The λ of AAV6 samples under four conditions: free, 100 mM Arg-HCl, 100 mM HP-β-CD, and 100 mM Arg-HCl + 100 mM HP-β-CD is shown. ＢＣＭ Figure 5-2 shows the results of Example 5. The λ of AAV6 samples was measured under four conditions: free, 100 mM Arg-Ace, 100 mM HP-β-CD, and 100 mM Arg-Ace + 100 mM HP-β-CD. ＢＣＭ Figure 5-3 shows the results of Example 5. The λ of AAV6 samples was measured under four conditions: free, 100 mM Arg-Suc, 100 mM HP-β-CD, and 100 mM Arg-Suc + 100 mM HP-β-CD. ＢＣＭ Figure 5-4 shows the results of Example 5. The λ values ​​of AAV6 samples were compared under four conditions: free, 100 mM Arg-Asp, 100 mM HP-β-CD, and 100 mM Arg-Asp + 100 mM HP-β-CD. ＢＣＭ The temperature dependence of the 2×10 １３vg / mL AAV6-CMV-EGFP, 10 mM His / HCl, 190 mM NaCl, 0.33% (w / v) PX188, pH 6.0 was used as the basic condition (190 mM NaCl), and 190 mM NaCl was used as the buffer solution for 190 mM Arg-Asp, 190 mM Arg-Asp + 20 mM MgCl ２ , 190mM Arg-Asp+20mM CaCl ２ , or 190 mM Arg-Asp + 100 mM sucrose. １３ vg / mL AAV6-CMV-EGFP, 10 mM His / HCl, 190 mM NaCl, 0.33% (w / v) PX188, pH 6.0 was used as the basic condition (190 mM NaCl), and 190 mM NaCl was used as the buffer solution for 190 mM Arg-Asp, 190 mM Arg-Asp + 20 mM MgCl ２ , 190mM Arg-Asp+20mM CaCl ２ , or 190 mM Arg-Asp + 100 mM sucrose ＢＣＭFIG. 7 shows the results of Example 7. ... Figure 9-1 shows a comparison of the temperature dependence of scattered light intensity of AAV6 samples under four conditions: free, 40 mM Arg-HCl, 40 mM HP-β-CD, and 40 mM Arg-HCl + 40 mM HP-β-CD. Figure 9-1 shows the results of Example 9. Figure 9-2 shows a comparison of the temperature dependence of scattered light intensity of AAV2 samples under four conditions: free, 100 mM Arg-HCl, 100 mM HP-β-CD, and 100 mM Arg-HCl + 100 mM HP-β-CD. Figure 9-3 shows the results of Example 9. Figure 9-4 shows a comparison of the temperature dependence of scattered light intensity of AAV8 samples under four conditions: free, 100 mM Arg-HCl, 100 mM HP-β-CD, and 100 mM Arg-HCl + 100 mM HP-β-CD. Figure 9-3 shows the results of Example 9. The temperature dependence of scattered light intensity of AAV9 samples was compared under four conditions: free, 100 mM Arg-HCl, 100 mM HP-β-CD, and 100 mM Arg-HCl + 100 mM HP-β-CD. Figure 10 shows the results of Example 10. The temperature dependence of scattered light intensity was compared under four conditions: 1.5 mg / mL insulin, 25 mM sodium phosphate, pH 8.0 as the basic condition (free), and 50 mM Arg-HCl, 50 mM HP-β-CD, or 50 mM Arg-HCl + 50 mM HP-β-CD added to the free condition. Figure 11-1 shows the results of Example 11.The temperature dependence of scattered light intensity was compared under four conditions: 1.5 mg / mL IgG, 5 mM Cit / NaOH, pH 6.0 (basic condition), and 100 mM NaCl, 100 mM HP-β-CD, or 100 mM NaCl + 100 mM HP-β-CD (free condition). Figure 11-2 shows the results of Example 11. The temperature dependence of scattered light intensity was compared under four conditions: 1.5 mg / mL IgG, 5 mM Cit / NaOH, pH 6.0 (basic condition), and 100 mM KCl, 100 mM HP-β-CD, or 100 mM KCl + 100 mM HP-β-CD (free condition). Figure 11-3 shows the results of Example 11. The basic conditions (free) were 1.5 mg / mL IgG, 5 mM Cit / NaOH, pH 6.0, and 100 mM MgCl . ２ , 100 mM HP-β-CD, or 100 mM MgCl ２ The temperature dependence of scattered light intensity was compared under four conditions, including the addition of 100 mM HP-β-CD. Figure 11-4 shows the results of Example 11. The basic condition (free) was 1.5 mg / mL IgG, 5 mM Cit / NaOH, pH 6.0, and the free condition was 100 mM CaCl. ２ , 100 mM HP-β-CD, or 100 mM CaCl ２Figure 11-5 shows the results of Example 11. The temperature dependence of scattered light intensity was compared under four conditions: 1.5 mg / mL IgG, 5 mM Cit / NaOH, pH 6.0 as the basic condition (free), and 100 mM Na-Ace, 100 mM HP-β-CD, or 100 mM Na-Ace + 100 mM HP-β-CD was added to the free condition. Figure 11-6 shows the results of Example 11. The temperature dependence of scattered light intensity was compared under four conditions: 1.5 mg / mL IgG, 5 mM Cit / NaOH, pH 6.0 (basic condition), and 100 mM Arg-HCl, 100 mM HP-β-CD, or 100 mM Arg-HCl + 100 mM HP-β-CD (free condition). Figure 11-7 shows the results of Example 11. The temperature dependence of scattered light intensity was compared under four conditions: 1.5 mg / mL IgG, 5 mM Cit / NaOH, pH 6.0 (basic condition), and 200 mM NaCl, 200 mM HP-β-CD, or 200 mM NaCl + 200 mM HP-β-CD (free condition). Figure 11-8 shows the results of Example 11. The temperature dependence of scattered light intensity was compared under four conditions: 1.5 mg / mL IgG, 5 mM Cit / NaOH, pH 6.0 (basic condition), and 200 mM Na-Ace, 200 mM HP-β-CD, or 200 mM Na-Ace + 200 mM HP-β-CD (free condition). Figure 11-9 shows the results of Example 11. The temperature dependence of scattered light intensity was compared under four conditions: 1.5 mg / mL IgG, 5 mM Cit / NaOH, pH 6.0 (basic condition), and 200 mM Arg-HCl, 200 mM HP-β-CD, or 200 mM Arg-HCl + 200 mM HP-β-CD (free condition). Figure 12-1 shows the results of Example 12. The temperature dependence of scattered light intensity was compared under four conditions: 1.5 mg / mL IgG, 5 mM Cit / NaOH, pH 6.0 (basal condition (free)), and 100 mM Arg-Ace, 100 mM HP-β-CD, or 100 mM Arg-Ace + 100 mM HP-β-CD added to the free condition.Figure 12-2 shows the results of Example 12. The temperature dependence of scattered light intensity was compared under four conditions: 1.5 mg / mL IgG, 5 mM Cit / NaOH, pH 6.0 (basic conditions), and 100 mM Arg-Suc, 100 mM HP-β-CD, or 100 mM Arg-Suc + 100 mM HP-β-CD (free conditions). Figure 12-3 shows the results of Example 12. The temperature dependence of scattered light intensity was compared under four conditions: 1.5 mg / mL IgG, 5 mM Cit / NaOH, pH 6.0 (basic conditions), and 100 mM Arg-Asp, 100 mM HP-β-CD, or 100 mM Arg-Asp + 100 mM HP-β-CD (free conditions). Figure 13-1 shows the results of Example 13. The temperature dependence of scattered light intensity was compared under four conditions: 1.5 mg / mL IgG, 5 mM Cit / NaOH, pH 6.0 (basic conditions), and 100 mM Arg-HCl, 100 mM HP-α-CD, or 100 mM Arg-HCl + 100 mM HP-α-CD (free conditions). Figure 13-2 shows the results of Example 13. The temperature dependence of scattered light intensity was compared under four conditions: 1.5 mg / mL IgG, 5 mM Cit / NaOH, pH 6.0 (basic conditions), and 100 mM Arg-HCl, 100 mM HP-γ-CD, or 100 mM Arg-HCl + 100 mM HP-γ-CD (free conditions). Figure 13-3 shows the results of Example 13. The temperature dependence of scattered light intensity was compared under four conditions: 1.5 mg / mL IgG, 5 mM Cit / NaOH, pH 6.0 (basic conditions), and 200 mM Arg-HCl, 200 mM HP-α-CD, or 200 mM Arg-HCl + 200 mM HP-α-CD (free conditions). Figure 13-4 shows the results of Example 13. The temperature dependence of scattered light intensity was compared under four conditions: 1.5 mg / mL IgG, 5 mM Cit / NaOH, pH 6.0 (basic conditions), and 200 mM Arg-HCl, 200 mM HP-γ-CD, or 200 mM Arg-HCl + 200 mM HP-γ-CD (free conditions). FIG. 14-1 shows the results of Example 14.The temperature dependence of scattered light intensity was compared under eight conditions: 1.5 mg / mL IgG, 5 mM Cit / NaOH, pH 6.0 (basic conditions), the Arg-HCl concentration was fixed at 100 mM, and the HP-β-CD concentration was varied from 0 to 200 mM. Figure 14-2 shows the results of Example 14. The temperature dependence of scattered light intensity was compared under eight conditions: 1.5 mg / mL IgG, 5 mM Cit / NaOH, pH 6.0 (basic conditions), the HP-β-CD concentration was fixed at 100 mM, and the Arg-HCl concentration was varied from 0 to 200 mM. Figure 14-3 shows the results of Example 14. The temperature dependence of scattered light intensity was compared under six conditions: the basic condition (free) was 1.5 mg / mL IgG, 5 mM Cit / NaOH, pH 6.0, and the molar concentration ratio of Arg-HCl to HP-β-CD was varied so that the total molar concentration of Arg-HCl and HP-β-CD was 200 mM. Figure 14-4 shows the results of Example 14. The temperature dependence of scattered light intensity was compared under eight conditions: the basic condition (free) was 1.5 mg / mL IgG, 5 mM Cit / NaOH, pH 6.0, the molar concentration ratio of Arg-HCl to HP-β-CD was 1:1, and the total molar concentration was varied from 120 mM (60 mM Arg-HCl + 60 mM HP-β-CD) to 400 mM (200 mM Arg-HCl + 200 mM HP-β-CD). 15 shows the results of Example 15. The results are shown for 10 mg / mL IgG, 5 mM Cit / NaOH, pH 6.0 (basal condition (free)), and the percentage (%) of soluble protein aggregates immediately after preparation (initial) and after 3 months of storage at 25°C under eight conditions (free) to which 100 mM Arg-HCl, 100 mM HP-β-CD, 50 mM Arg-HCl + 50 mM HP-β-CD, 200 mM Arg-HCl, 200 mM HP-β-CD, 100 mM Arg-HCl + 100 mM HP-β-CD, or 200 mM Arg-HCl + 200 mM HP-β-CD were added. The ratio (%) of the peak area of ​​the dimer or higher fraction to the total peak area was evaluated as the ratio (%) of soluble protein aggregates (Aggregates (%)). Figure 16-1 shows the results of Example 16.The basic conditions (free) were 10 mg / mL IgG, 5 mM Cit / NaOH, pH 6.0, and 100 mM Arg-HCl, 100 mM HP-β-CD, 50 mM Arg-HCl + 50 mM HP-β-CD, 200 mM Arg-HCl, 200 mM HP-β-CD, 100 mM Arg-HCl + 100 mM HP-β-CD, or 200 mM Arg-HCl + 200 mM HP-β-CD. The results are shown below. The particle concentration (Particles / mL) of >2 μm size was evaluated by flow imaging (FI) immediately after preparation (initial) and after 3 months of storage at 25 ° C. under eight conditions. Four measurements were performed, and the average and standard deviation of the second, third, and fourth measurement data were calculated. The particle size was determined as the area-based diameter (ABD). Figure 16-2 shows the results of Example 16. The basic conditions (free) were 10 mg / mL IgG, 5 mM Cit / NaOH, pH 6.0, and 100 mM Arg-HCl, 100 mM HP-β-CD, 50 mM Arg-HCl + 50 mM HP-β-CD, 200 mM Arg-HCl, 200 mM HP-β-CD, 100 mM Arg-HCl + 100 mM HP-β-CD, or 200 mM Arg-HCl + 200 mM HP-β-CD. The particle concentration (Particles / mL) of > 5 μm size was evaluated by flow imaging (FI) immediately after preparation (initial) and after 3 months of storage at 25 ° C. The results are shown. Four measurements were performed, and the average and standard deviation of the second, third, and fourth measurement data were calculated. The particle size was determined as the area-based diameter (ABD). Figure 16-3 shows the results of Example 16.The basic conditions (free) were 10 mg / mL IgG, 5 mM Cit / NaOH, pH 6.0, and 100 mM Arg-HCl, 100 mM HP-β-CD, 50 mM Arg-HCl + 50 mM HP-β-CD, 200 mM Arg-HCl, 200 mM HP-β-CD, 100 mM Arg-HCl + 100 mM HP-β-CD, or 200 mM Arg-HCl + 200 mM HP-β-CD. The particle concentration (Particles / mL) of >10 μm size was evaluated by flow imaging (FI) immediately after preparation (initial) and after 3 months of storage at 25 ° C. The results are shown. Four measurements were performed, and the average and standard deviation of the second, third, and fourth measurement data were calculated. The particle size was determined as the area-based diameter (ABD). Figure 16-4 shows the results of Example 16. The basic conditions (free) were 10 mg / mL IgG, 5 mM Cit / NaOH, pH 6.0, and 100 mM Arg-HCl, 100 mM HP-β-CD, 50 mM Arg-HCl + 50 mM HP-β-CD, 200 mM Arg-HCl, 200 mM HP-β-CD, 100 mM Arg-HCl + 100 mM HP-β-CD, or 200 mM Arg-HCl + 200 mM HP-β-CD. The particle concentration (Particles / mL) of >25 μm size was evaluated by flow imaging (FI) immediately after preparation (initial) and after 3 months of storage at 25 ° C. The results are shown. Four measurements were performed, and the average and standard deviation of the second, third, and fourth measurement data were calculated. The particle size was determined as the area-based diameter (ABD).

[0011] Throughout this specification, singular expressions should be understood to include the plural concept unless otherwise specified. Thus, singular articles (e.g., "a," "an," "the," etc. in English) should be understood to include the plural concept unless otherwise specified. Furthermore, it should be understood that terms used in this specification are used in the sense commonly used in the art unless otherwise specified. Therefore, unless otherwise defined, all technical and scientific terms used in this specification have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. In case of conflict, the present specification (including definitions) will control.

[0012] (Definitions) We first explain the terms and general techniques used in this disclosure.

[0013] As used herein, the term "protein (component)" is used in the same sense as commonly used in the art to refer to a polymer of amino acids, which is the subject of the present disclosure. Preferably, a "polypeptide" or "protein" is a sequence of amino acids whose chain length is sufficient to form a tertiary structure; proteins herein are distinguished from "peptides," which are short amino acid molecules that generally lack any tertiary structure. Typically, proteins as used herein have a molecular weight of at least about 5-20 kD, alternatively at least about 15-20 kD, preferably at least about 20 kD. Polypeptides or proteins herein include, for example, antibodies. As used herein, protein includes or is used interchangeably with other terms referring to polymers of amino acids, such as polypeptides, oligopeptides, and peptides (when intended to have a three-dimensional structure), whether modified or unmodified.

[0014] Examples of polypeptides encompassed within the definition herein include mammalian proteins, such as various antibodies, renin; growth hormones, including human growth hormone and bovine growth hormone; growth hormone-releasing factor; parathyroid hormone; thyroid-stimulating hormone; lipoproteins; alpha-1-antitrypsin; insulin A chain; insulin B chain; proinsulin; follicle-stimulating hormone; calcitonin; luteinizing hormone; glucagon; clotting factors, such as factor VIIIC, factor IX, tissue factor, and von Willebrand factor; anticoagulants, such as ribonucleotides (RIs), ribonucleotides (RIs), and ribonucleotides (RIs). for example, protein C; atrial natriuretic factor; pulmonary surfactant; plasminogen activators, such as urokinase or human urinary or tissue-type plasminogen activator (t-PA); bombesin; thrombin; hematopoietic growth factors; tumor necrosis factor-alpha and -beta; enkephalinase; RANTES (regulated upon activation and normally expressed and secreted by T cells); human macrophage inflammatory protein (MIP-1-alpha); serum albumin, for example, human serum albumin; Müllerian inhibitory substance; relaxin A chain; relaxin a cytotoxic T-lymphocyte-associated antigen (CTLA), e.g., CTLA-4; inhibin; activin; vascular endothelial growth factor (VEGF); a hormone or growth factor receptor; protein A or D; rheumatoid factor; a neurotrophic factor, e.g., bone-derived neurotrophic factor (BDNF), neurotrophin-3, neurotrophin-4, neurotrophin-5, or neurotrophin-6 (NT-3, NT-4, NT-5, or NT-6), or nerve growth factors, such as NGF-β; platelet-derived growth factor (PDGF); fibroblast growth factors, such as aFGF and bFGF; epidermal growth factor (EGF); transforming growth factors (TGFs), such as TGF-alpha and TGF-beta, including TGF-β1, TGF-β2, TGF-β3, TGF-β4, or TGF-β5; insulin-like growth factors I and II (IGF-I and IGF-II); des(1-3)-IGF-I (brain IGF-I), insulin-like growth factor binding proteins (IGFBPs);CD proteins, such as CD3, CD4, CD8, CD19, and CD20; erythropoietin; osteoinductive factors; immunotoxins; bone morphogenetic proteins (BMPs); interferons, such as interferon-alpha, interferon-beta, and interferon-gamma; colony-stimulating factors (CSFs), such as M-CSF, GM-CSF, and G-CSF; interleukins (ILs), such as IL-1 to IL-10; superoxide dismutase; T-cell receptors; surface membrane proteins; decay-accelerating factors; viral antigens, such as AIDS envelope Examples of proteins include fragments and / or variants of any of the above proteins, as well as antibodies that are antibody fragments and bind to any of the above proteins; coat proteins, such as various viral capsids; transport proteins; homing receptors; addressins; regulatory proteins; integrins, such as CD11a, CD11b, CD11c, CD18, ICAM, VLA-4, and VCAM; tumor-associated antigens, such as CA125 (ovarian cancer antigen), or HER2, HER3, or HER4 receptors; immunoadhesins; and fragments and / or variants of any of the above proteins, as well as antibodies that are antibody fragments and bind to any of the above proteins; coat proteins, such as capsids, of various viruses;

[0015] Proteins herein broadly encompass naturally occurring proteins, as well as fusion proteins formed, for example, by covalently linking two separate proteins to one another, composite proteins formed by non-covalently linking two or more separate proteins to one another, and protein conjugates, which include proteins covalently linked to other proteins or non-protein molecules such as nucleic acids, small molecule drugs, or solid phases.

[0016] A protein retains "biological activity" in a pharmaceutical formulation if the biological activity of the protein at a given time is within about 10% (within the error of the assay) of the biological activity exhibited at the time the formulation was prepared. In the case of antibodies or proteins intended to function by binding to a target molecule or antigen, biological activity can be determined by the ability of the protein to bind to the antigen and produce a measurable biological response in vitro or in vivo.

[0017] As used herein, a protein is "stable" if the protein contained therein essentially retains its physical and / or chemical stability during storage. Stability can be confirmed by the methods used in the Examples, or alternatively, can be measured at a selected temperature for a selected period of time. When the protein is an antibody, preferably, the formulation is stable at room temperature (about 25°C) or 40°C for at least one month, and / or at about 2-8°C for at least one year, preferably at least two years. When the protein is a capsid, preferably, the formulation is stable at room temperature (about 25°C) for at least one month, and / or at about 2-8°C for at least one year, preferably at least two years, and / or at about 0°C or below for at least one year, preferably at least two years. For example, the degree of aggregation during storage can be used as an indicator of protein stability. Thus, a "stable" formulation may be one in which the percentage of soluble protein aggregates (calculated as the ratio of the peak area of ​​aggregates to the total peak area when analyzed by size exclusion chromatography) increases by less than about 10% during storage, and preferably less than 5%, 3%, or 2%. Various analytical techniques for measuring protein stability are available in the art and are reviewed, for example, in Mahler HC, Friess W, Grauschopf U, Kiese S. Protein aggregation: pathways, induction factors and analysis. J Pharm Sci. 2009;98(9):2909-2934.

[0018] Increasing the "stability" of a protein-containing formulation can include reducing or preventing the formation of protein aggregates or degradation products of other components of the formulation (compared to an untreated protein-containing formulation) in the formulation, so that those other components can continue to act to maintain protein stability.

[0019] As used herein, the term "electrolyte" refers to a nonmetallic electrical conductor that carries electric current by the movement of ions, or a substance that becomes an ionic conductor when dissolved in a solvent. In the present disclosure, the term mainly refers to substances that generate ions when dissolved in a solvent such as water, and the electrolyte contained in the composition in the present disclosure may be applied in the state of an electrolyte or may be contained in the state of its ions.

[0020] As used herein, the term "aggregation" as used herein means coming together or gathering into a mass or whole, such as, for example, the aggregation of protein molecules.

[0021] As used herein, "aggregate" refers to an association of at least two molecules, and often more (e.g., 5, 10, 20, or more) molecules. The association may be either covalent or non-covalent with respect to the mechanism by which the molecules associate. The association may be direct between the molecules or indirect through other molecules that link the antibodies to one another. In some embodiments, the aggregated molecules comprise aggregated target molecules. For example, in some embodiments, the aggregate comprises aggregated antibodies. The aggregate may be self-aggregating, or may aggregate due to the presence of other factors, such as aggregating agents, precipitants, agitation, or other means and methods by which proteins come together. An "aggregation-prone" protein is one that has been observed to aggregate with other protein molecules, particularly upon agitation. Aggregation can be observed visually, such as when a previously clear protein formulation in solution becomes cloudy or contains a precipitate, or by methods such as size exclusion chromatography (SEC), which separates proteins in a formulation by size; flow imaging, dynamic light scattering, or light obscuration, which weigh particles in a formulation; and multi-angle light scattering or mass photometry, which measure molecular weight. Aggregates can include dimers, trimers, and multimers of protein species. As used herein, "high molecular weight species" (also referred to as "Aggregates (%)" in this example) refers to protein aggregates that can be observed, for example, by size exclusion chromatography and represent at least dimers of the desired protein molecule, i.e., have at least twice the molecular weight of the desired protein species in the formulation. For protein species such as antibodies that are already multimeric in their normal or desired form, e.g., dimers or tetramers, "Aggregates (%)" represents at least dimers of the normal, desired multimeric form of the protein.

[0022] As used herein, the term "organic electrolyte" refers to an electrolyte in which at least one of the constituent acids, bases, and salts is organic.

[0023] As used herein, the term "inorganic electrolyte" refers to an electrolyte in which all of the constituent acids, bases, and salts are inorganic.

[0024] As used herein, the term "inorganic salt" generally refers to a type of inorganic compound, in which the constituent base and acid are both inorganic. Therefore, even if the constituent base or acid is inorganic, if the remaining constituents include an organic acid or organic base, it is included in the category of inorganic salt. These salts may be produced by the reaction of an acid and a base, by the direct bonding of a metal and a non-metal, or by other chemical means. In a broad sense, inorganic salts include the forms of naturally occurring minerals and industrially synthesized compounds. Examples include various inorganic salts such as sodium chloride (NaCl), copper sulfate (CuSO ４ ), calcium carbonate (CaCO ３ ), potassium nitrate (KNO ３ ), ammonium chloride (NH ４ Cl), ammonium sulfate ((NH ４ ) ２ SO ４ ), magnesium chloride (MgCl ２ ), sodium fluoride (NaF), calcium phosphate (Ca ３ (P.O. ４ ) ２ ), ammonium perchlorate (NH ４ ClO ４ ), potassium chloride (KCl), sodium sulfate (Na ２ SO ４ ), iron chloride (FeCl ３ ), silver nitrate (AgNO ３ ), barium sulfate (BaSO ４ ), calcium chloride (CaCl ２ ), zinc sulfate (ZnSO ４ ), sodium carbonate (Na ２ CO ３ ), magnesium sulfate (MgSO ４ ), sodium phosphate (Na ３ P.O. ４ ), zinc chloride (ZnCl ２ ), potassium sulfate (K ２ SO ４ ), lithium carbonate (Li ２ CO ３ ), lead nitrate (Pb(NO ３ ) ２ ), copper chloride (CuCl２ ), aluminum sulfate (Al ２ (SO ４ ) ３ ), strontium nitrate (Sr(NO ３ ) ２ ), calcium fluoride (CaF ２ ), lithium chloride (LiCl), etc.

[0025] As an example, the inorganic salt in this specification is composed of an inorganic acid such as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, or carbonic acid, and an inorganic base based on an alkali metal, alkaline earth metal, or the like. Examples include alkali metal salts, alkaline earth metal salts, and ammonium salts of inorganic acids such as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, and carbonic acid. Specifically, for example, one or more salts selected from the group consisting of sodium chloride, potassium chloride, ammonium chloride, calcium chloride, magnesium chloride, sodium sulfate, potassium sulfate, magnesium sulfate, sodium nitrate, potassium nitrate, magnesium nitrate, ammonium nitrate, disodium monohydrogen phosphate, sodium dihydrogen phosphate, trisodium potassium monohydrogen phosphate, potassium dihydrogen phosphate, tripotassium phosphate, sodium bicarbonate, and calcium carbonate can be selected and used.

[0026] As used herein, the term "organic salt" generally refers to any salt composed of an organic substance. If either the constituent acid or base contains an organic substance, it falls under the category of organic salt in the present disclosure. For example, it is generally produced by the neutralization reaction of an organic acid with a base (such as a metal hydroxide or ammonia). Organic salts are widely found in nature. Examples of "organic substances" include salts with formic acid, acetic acid, fumaric acid, oxalic acid, tartaric acid, maleic acid, lactic acid, citric acid, succinic acid, malic acid, benzoic acid, glutamic acid, and aspartic acid. Among these, acetic acid, citric acid, tartaric acid, glutamic acid, succinic acid, malic acid, and aspartic acid are preferred, and salts with acetic acid, succinic acid, and aspartic acid are more preferred. The organic salt may contain an organic base. Examples of "organic bases" include arginine, histidine, and tris(hydroxymethyl)aminomethane, with arginine being preferred. "Organic electrolytes" include, for example, organic bases formed by combining an amine or other nitrogen-containing organic compound with an inorganic or organic acid. The organic base salts contain at least one organic moiety (including a carbon chain or ring structure), an organic base that functions as a cation, and the corresponding acid anion. For example, the following organic base salts exist: pyridinium hydrochloride (C ５ H ５ NHCl), triethylammonium acetate ((C ２ H ５ ) ３ N HAc), dimethylanilinium sulfate ((C ６ H ５ NH (CH ３ ) ２ ) ２ SO ４ ), benzethonium hydrochloride (C ２７ H ４２ ClNO ２ ), methyltriphenylphosphonium bromide ((C ６ H ５ ) ３ PCH ３ Br), tetramethylammonium acetate ((CH ３ ) ４ N HAc), tetrabutylammonium sulfate ((C ４ H ９ ) ４NHSO ４ ), hydroxylamine hydrochloride (NH ２ OH·HCl), diethylaminoethanol hydrochloride ((C ２ H ５ ) ２ NCH ２ CH ２ OH·HCl), hexadecyltrimethylammonium bromide (C １９ H ４２ NBr) etc.

[0027] Examples of organic salts used in the present disclosure include lithium formate, lithium acetate, lithium fumarate, lithium oxalate, lithium tartrate, lithium maleate, lithium lactate, lithium citrate, lithium succinate, lithium malate, lithium benzoate, sodium formate, sodium acetate, sodium fumarate, sodium oxalate, sodium tartrate, sodium maleate, sodium lactate, sodium citrate, sodium succinate, sodium malate, sodium benzoate, potassium formate, potassium acetate, potassium fumarate, potassium oxalate, potassium tartrate, potassium maleate, potassium lactate, potassium citrate, potassium succinate, potassium malate, potassium benzoate, magnesium stearate, magnesium citrate, and the like.

[0028] As used herein, the term "pharmaceutically acceptable" refers to substances commonly used in the technical field of pharmaceutical formulations, such as excipients, binders, disintegrants, emulsifiers, flow additives, lubricants, stabilizers, tonicity agents, buffers, pH adjusters, solubilizers, preservatives, surfactants, etc.

[0029] As used herein, "pharmaceutically acceptable acids" include inorganic and organic acids that are non-toxic at the concentrations and manners in which they are formulated. For example, suitable inorganic acids include hydrochloric acid, perchloric acid, hydrobromic acid, hydroiodic acid, nitric acid, sulfuric acid, sulfonic acid, sulfinic acid, sulfanilic acid, phosphoric acid, carbonic acid, etc. Suitable organic acids include straight and branched chain alkyl, aromatic, cyclic, alicyclic, arylaliphatic, heterocyclic, saturated, unsaturated mono-, di-, and tricarboxylic acids (e.g., formic acid, acetic acid, 2-hydroxyacetic acid, trifluoroacetic acid, phenylacetic acid, trimethylacetic acid, t-butylacetic acid, anthranilic acid, propanoic acid, 2-hydroxypropanoic acid, 2-oxopropanoic acid, propanedioic acid, cyclopentanepropionic acid, cyclopentanepropionic acid, 3-phenylpropionic acid, butanoic acid, butanedioic acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, 2-acetoxybenzoic acid, ascorbic acid, cinnamic acid, lauryl sulfuric acid, stearic acid, muconic acid, mandelic acid, succinic acid, embonic acid, fumaric acid, malic acid, Included are maleic acid, hydroxymaleic acid, malonic acid, lactic acid, citric acid, tartaric acid, glycolic acid, glyconic acid, gluconic acid, pyruvic acid, glyoxalic acid, oxalic acid, mesylic acid, succinic acid, salicylic acid, phthalic acid, palmoic acid, palmaic acid, thiocyanic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-colobenzenesulfonic acid, naphthalene-2-sulfonic acid, p-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylic acid, glucoheptonic acid, 4,4'-methylenebis-3-(hydroxy-2-ene-1-carboxylic acid), and hydroxynaphthoic acid.

[0030] As used herein, "pharmaceutically acceptable base" includes inorganic and organic bases that are non-toxic at the concentrations and in the manner in which they are formulated. For example, suitable bases include those formed from metals that form inorganic bases, such as lithium, sodium, potassium, magnesium, calcium, ammonium, iron, zinc, copper, manganese, aluminum, and the like; N-methylglucamine, morpholine, piperidine, and organic non-toxic bases (including primary, secondary, and tertiary amines, substituted amines, cyclic amines, and basic ion exchange resins, [e.g., N(R')4+, where R' is independently H or C1-4 alkyl, e.g., ammonium, tris]) such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-diethylaminoethanol, trimethamine, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, methylglucamine, theobromine, purines, piperazine, piperidine, N-ethylpiperidine, polyamine resins, and the like). Particularly preferred organic non-toxic bases are isopropylamine, diethylamine, ethanolamine, trimethamine, dicyclohexylamine, choline, and caffeine.

[0031] Additional pharmaceutically acceptable acids and bases that can be used in the present disclosure include those derived from amino acids such as arginine, histidine, glycine, phenylalanine, aspartic acid, glutamic acid, lysine, and asparagine.

[0032] As used herein, the term "cyclic oligosaccharide" refers to any oligosaccharide in a cyclic form, and examples thereof include cyclodextrin, cyclomannin, cycloaltrin, cyclofructin, etc. Cyclodextrin is preferred, but is not limited to this.

[0033] As used herein, the term "cyclodextrin" refers to a dextrin having a ring structure. It typically refers to a natural, unsubstituted cyclodextrin containing 6 to 12 glucose units linked by a covalent bond between carbon 1 and carbon 4. Typically, cyclodextrins contain 6, 7, or 8 glucose residues in the ring, forming a cone shape, and are represented as follows: α (alpha)-cyclodextrin: a 6-membered sugar ring molecule β (beta)-cyclodextrin: a 7-membered sugar ring molecule γ (gamma)-cyclodextrin: an 8-membered sugar ring molecule

[0034] α-CD was used in a powder formulation (HypoGon® Nasal) by Novo Nordisk in clinical trials (Stenniger et al., Diabetologia 1993;36:931-935; Rosenfalck AM, et al., Diabetes Res Clin Pract 1992;17:43-50). The aqueous solubility of α-CD is reported to be approximately 5% by weight. Two other cyclodextrins, one less soluble in water than α-CD (β-CD, 1.85% by weight) and one more soluble in water than α-CD (HP-β-CD), are also suitable for use in the present disclosure, as is γ-cyclodextrin, which is freely soluble in water.

[0035] Cyclodextrins may be used individually or as a mixture of any two or more cyclodextrins, which can be produced, for example, by the intramolecular transfer reaction of α-1,4-glucan such as starch with cyclodextrin glucanotransferase.

[0036] In this specification, the term "cyclodextrin" may refer to its derivatives, which are referred to as cyclodextrin derivatives (also referred to as CD derivatives). The CD derivatives used in the present disclosure have a structure in which an amino group of cyclodextrin is modified and the amino group is bonded to a carboxyl group at the 6-carbon position of a uronic acid (e.g., glucuronic acid) via an amide bond. Specifically, the CD derivatives represented by the following general formula (1)

[0037]

[0038] [In the above general formula (1), m is 0 to 7, n is 1 to 8, and m+n=6 to 8; R １ , R ２ , R ３ , R ４ , R ５ are each independently R ２ or R ４ one of the groups represents a group represented by -H and the other represents -OH or a group represented by the following formula (a), and R ３ or R ５ one of the groups represents a group represented by -H and the other represents -OH or a group represented by the following formula (a), and R １ represents —OH or a group represented by the following formula (a), and at least one of the following formula (a) is present.

[0039] -NH-Z...(a)

[0040] [In the formula, Z represents a residue of a monosaccharide or oligosaccharide having a reducing group, which is formed by condensing a uronic acid or an oligosaccharide containing a uronic acid with an amino group.]

[0041] This structure results in the 1-carbon atom of the uronic acid (e.g., glucuronic acid) that forms the branched (sugar-modified) structure becoming an aldehyde group, thereby exhibiting reducing properties. In this specification, cyclodextrin derivatives having this structure may be referred to as "sugar-modified cyclodextrins having a reducing end," etc. The branched structure may be imparted to, for example, α-CD, β-CD, or γ-CD. Since each CD has different properties, one may be selected appropriately depending on the desired properties.

[0042] In this specification, "equivalent molar amounts of cyclodextrin and salt" refers to calculations based on the molar amount of cyclodextrin itself when a cyclodextrin derivative is used, and calculations based on the molar amount of cyclodextrin itself when an electrolyte is used. Furthermore, when a divalent or trivalent ion is used, calculations are based on the molar amount after ionization. The molecular weight is the weight-average molecular weight.

[0043] In this specification, "carbohydrate" refers to a general term for carbohydrates, broadly classified into monosaccharides, disaccharides, and polysaccharides. Carbohydrates are carbon compounds containing hydrogen, oxygen, and water in proportions, and their structures and functions are extremely diverse. Monosaccharides are the most basic units of carbohydrates, and they combine to form disaccharides and polysaccharides. Carbohydrates are also important components of foods, and exist in a wide range of forms, from naturally occurring to artificially processed forms. Examples of carbohydrates include: monosaccharides (the simplest sugars, such as glucose (blood sugar), fructose, and galactose), disaccharides (combined pairs of monosaccharides, such as sucrose, lactose, and maltose), polysaccharides (combined pairs of monosaccharides, such as cellulose, starch, glycogen, and chitin), and other carbohydrates (oligosaccharides (carbohydrates consisting of a small number of monosaccharides), dextrins, and their cyclic forms).

[0044] In this specification, the term "surfactant" refers to a substance having both hydrophilic and hydrophobic groups in the molecule, and surfactants include ionic surfactants and nonionic surfactants. The term "ionic surfactant" refers to an ionic surfactant that, when dissolved in water, dissociates to form ions (charged atoms or atomic groups). Ionic surfactants are further classified into anionic surfactants, cationic surfactants, and amphoteric surfactants depending on the charge of the ions produced.

[0045] Examples of nonionic surfactants include sugar ester surfactants such as sorbitan fatty acid esters (C12-18), POE sorbitan fatty acid esters (C12-18), and sucrose fatty acid esters; fatty acid ester surfactants such as POE fatty acid esters (C12-18), POE resin acid esters, and POE fatty acid diesters (C12-18); alcohol surfactants such as POE alkyl ethers (C12-18); alkylphenol surfactants such as POE alkyl (C8-12) phenyl ether, POE dialkyl (C8-12) phenyl ether, and POE alkyl (C8-12) phenyl ether formalin condensates; polyoxyethylene-polyoxypropylene block polymers; Examples of surfactants include polyoxyethylene-polyoxypropylene block polymer surfactants such as alkyl (C12-18) polyoxyethylene-polyoxypropylene block polymer ethers; alkylamine surfactants such as POE alkylamine (C12-18) and POE fatty acid amide (C12-18); bisphenol surfactants such as POE fatty acid bisphenyl ether; polyaromatic ring surfactants such as POA benzylphenyl (or phenylphenyl) ether and POA styrylphenyl (or phenylphenyl) ether; POE ether and ester type silicone and fluorine-based surfactants; and vegetable oil surfactants such as POE castor oil and POE hydrogenated castor oil. Preferred surfactants include polyoxyl 40 stearate, sorbitan trioleate, polyoxyethylene (105) polyoxypropylene (5) glycol, polyoxyethylene hydrogenated castor oil 60, polyoxyl 35 castor oil, and lauromacrogol.

[0046] Anionic surfactants include alkyl sulfates (C12-18, Na, NH ４ , alkanolamine), POE alkyl ether sulfate (C12-18, Na, NH ４ , alkanolamine), POE alkyl phenyl ether sulfate (C12-18, NH ４ , alkanolamine, Ca), POE benzyl (or styryl) phenyl (or phenylphenyl) ether sulfate (Na, NH ４, alkanolamine), polyoxyethylene, polyoxypropylene block polymer sulfate (Na, NH ４ , alkanolamine); paraffin (alkane) sulfonate (C12-22, Na, Ca, alkanolamine), AOS (C14-16, Na, alkanolamine), dialkyl sulfosuccinate (C8-12, Na, Ca, Mg), alkyl benzene sulfonate (C12, Na, Ca, Mg, NH ４ , alkylamines, alkanols, amines, cyclohexylamine), mono- or di-alkyl (C3-6) naphthalene sulfonates (Na, NH ４ , alkanolamine, Ca, Mg), naphthalenesulfonate-formalin condensate (Na, NH ４ ), alkyl (C8-12) diphenyl ether disulfonate (Na, NH ４ ), lignin sulfonate (Na, Ca), POE alkyl (C8-12) phenyl ether sulfonate (Na), POE alkyl (C12-18) ether sulfosuccinic acid half ester (Na); ４, alkanolamine), N-methyl-fatty acid sarcosinate (C12-18, Na), resinate (Na, K), and other carboxylic acid surfactants; POE alkyl (C12-18) ether phosphate (Na, alkanolamine), POE mono- or dialkyl (C8-12) phenyl ether phosphate (Na, alkanolamine), POE benzyl (or styryl) phenyl (or phenylphenyl) ether phosphate (Na, alkanolamine), polyoxyethylene-polyoxypropylene block polymer (Na, alkanolamine), phosphatidylcholine-phosphatidylethanolimine (lecithin), alkyl (C8-12) phosphate, and other phosphate surfactants. Preferred surfactants include monoalkyl sulfates such as sodium lauryl sulfate, sodium tetradecyl sulfate, sodium hexadecyl sulfate, and sodium octadecyl sulfate; dioctyl sodium sulfosuccinate, sodium lauroyl sarcosinate, and sodium dodecylbenzenesulfonate.

[0047] In the present specification, two or more surfactants may be used in combination in an appropriate ratio, and the surfactant content of the composition is 0.001% by weight or more, preferably 0.001% by weight or more and 5% by weight or less, more preferably 0.005% by weight or more, and most preferably 0.005% by weight or more and 1% by weight or less, based on the total weight of the composition.

[0048] As used herein, a "vial" refers to a container that is filled with a liquid and can store the liquid by sealing it. Examples of such a vial include plastic or glass containers.

[0049] As used herein, a "syringe" refers to a container that is filled with a liquid and has a needle attached thereto so that the liquid can be administered into the body or transferred to another container such as a vial. Examples of such a container include plastic or glass containers, and they can also be used as liquid storage containers.

[0050] As used herein, the term "kit" refers to any manufactured product (e.g., a package or container) containing at least one reagent, such as the pH buffer solution of the present disclosure. The kit can be promoted, distributed, or sold as a means for practicing the methods of the present disclosure. Furthermore, the kit can include a package insert explaining the kit and its use. Any or all of the kit reagents can be provided in a container that protects them from the external environment, such as a sealed container or pouch. As used herein, "kit" refers to a unit in which the components to be provided (e.g., test reagents, diagnostic reagents, therapeutic agents, antibodies, labels, instructions, etc.) are provided, typically separated into two or more compartments. This kit format is preferred when the purpose is to provide a composition that, for stability reasons, should not be provided in a mixed state, but is preferably mixed immediately before use. Such kits advantageously include instructions or manuals describing how to use the components provided (e.g., test reagents, diagnostic reagents, therapeutic agents) or how to handle the reagents. When the "kit" is used herein as a reagent kit, the kit typically includes instructions describing how to use the test reagents, diagnostic reagents, therapeutic agents, antibodies, etc.

[0051] As used herein, "concentration" refers to a process or operation of increasing the concentration of a target substance by reducing the amount of liquid in which the target substance is dispersed. The concentration is increased by at least 2 times, preferably at least 10 times, more preferably at least 100 times, and even more preferably at least 1000 times the original concentration. For example, methods such as ultrafiltration, dialysis, tangential flow filtration, concentration columns, centrifugation, and drying are used.

[0052] As used herein, a "lyophilized" formulation is a formulation that has been freeze-dried or subjected to a freeze-drying process. The formulations herein may be freeze-dried for storage or may be intended for storage as a liquid solution. A "reconstituted" formulation is one prepared by dissolving a freeze-dried protein or antibody formulation in a diluent so that the protein is dispersed in the reconstituted formulation. The reconstituted formulation may be suitable for use, such as administration to a patient to be treated with the protein of interest.

[0053] When the formulation is freeze-dried, a lyoprotectant may be added. A "lyoprotectant" is a molecule that, when combined with a protein of interest, significantly prevents or reduces the physicochemical instability of the protein during freeze-drying and subsequent storage. Exemplary lyoprotectants include sugars and their corresponding sugar alcohols; amino acids, such as monosodium glutamate or histidine; methylamines, such as betaine; lyotropic salts, such as magnesium sulfate; polyols, such as trihydric or higher molecular weight sugar alcohols, for example, glycerin, dextran, erythritol, glycerol, arabitol, xylitol, sorbitol, and mannitol; propylene glycol; polyethylene glycol; Pluronics®; and combinations thereof. Further exemplary lyoprotectants include glycerin and gelatin, and the sugars melibiose, melezitose, raffinose, mannotriose, and stachyose. Examples of reducing sugars include glucose, maltose, lactose, maltulose, isomaltulose, and lactulose. Examples of non-reducing sugars include non-reducing glycosides of polyhydroxy compounds selected from sugar alcohols and other linear polyhydric alcohols. Preferred sugar alcohols are monoglycosides, particularly compounds obtained by reduction of disaccharides such as lactose, maltose, lactulose, and maltulose. The glycoside side chain can be either a glucoside or a galactoside. Further examples of sugar alcohols are glucitol, maltitol, lactitol, and isomaltulose. Preferred lyoprotectants are the non-reducing sugars trehalose or sucrose.

[0054] As used herein, "surfactants" are molecules with well-defined polar and non-polar regions that allow them to aggregate in solution to form micelles. Depending on the nature of the polar regions, surfactants can be nonionic, anionic, cationic, and zwitterionic.

[0055] For purposes herein, a "diluent" is one that is pharmaceutically acceptable (safe and non-toxic for human administration) and useful for the preparation of a liquid formulation, e.g., a formulation to be reconstituted after lyophilization. Exemplary diluents include sterile water, bacteriostatic water for injection (BWFI), a pH buffered solution (e.g., phosphate-buffered saline), sterile saline solution, Ringer's solution, or dextrose solution. In alternative embodiments, the diluent may comprise an aqueous salt solution and / or buffer.

[0056] As used herein, percentages ("%") are weight to volume ("w / v") percentages unless otherwise specified.

[0057] As used herein, the term "consisting essentially of" when referring to a mixture of formulation components indicates that components other than those explicitly listed may be present, but that such components are found only in trace amounts, or otherwise in amounts sufficiently low, such that the basic characteristics of the formulation, including protein concentration, level of protein aggregation, level of protein oxidation, viscosity, thermal stability, osmolality, and pH, are unchanged.

[0058] As used herein, "AAV" refers to an adeno-associated virus. An "AAV capsid" refers to a protein particle containing the AAV outer protein capsid, and includes both particle forms containing nucleic acid within the capsid (e.g., adeno-associated virus vectors and AAV drug substances) and particle forms not containing nucleic acid. The capsid is composed of three polypeptides, VP1, VP2, and VP3, and typically has a molecular weight of at least about 3,000-4,000 kD, or at least about 3,500-4,000 kD, preferably at least about 3,500 kD.

[0059] (Preferred Embodiments) A description of preferred embodiments will be given below, but it should be understood that this embodiment is an example of the present disclosure and that the scope of the present disclosure is not limited to such preferred embodiments. It should also be understood that those skilled in the art can easily make modifications, changes, etc. within the scope of the present disclosure by referring to the following preferred examples. Therefore, it is clear that those skilled in the art can make appropriate modifications within the scope of the present disclosure by taking into account the description in this specification. It should also be understood that the following embodiments of the present disclosure can be used alone or in combination.

[0060] (Protein Preservation) In one aspect, the present disclosure provides a composition for preserving proteins, comprising an electrolyte and / or an ion thereof and a cyclic oligosaccharide or a derivative thereof. Without wishing to be limited thereto, it is an important point of the present disclosure that a combination of an electrolyte and / or an ion thereof and a cyclic oligosaccharide or a derivative thereof is used as such a protein preservative.

[0061] In another aspect, the present disclosure provides a composition comprising a protein, an electrolyte and / or ions thereof, and a cyclic oligosaccharide or derivative thereof, which is used to preserve the protein, since the protein is more effectively preserved in the composition than if the other components were not present.

[0062] In one embodiment, the electrolyte used in the present disclosure comprises an organic electrolyte.

[0063] In another embodiment, the electrolyte used in the present disclosure comprises one or more selected from inorganic salts, organic salts, organic acids, and organic bases.

[0064] In a preferred embodiment, the electrolyte used in the present disclosure comprises at least one organic acid and at least one organic base.

[0065] The compositions of the present disclosure may be solutions or solids. In one embodiment, the compositions of the present disclosure are solutions. In another embodiment, the compositions of the present disclosure are solids.

[0066] The electrolytes used in the present disclosure are typically present at 100 millimoles or more per liter of the total composition of the present disclosure. In certain embodiments, the electrolytes of the present disclosure can be, for example, 10 millimoles or more, 20 millimoles or more, 30 millimoles or more, 40 millimoles or more, 50 millimoles or more, 60 millimoles or more, 70 millimoles or more, 80 millimoles or more, 90 millimoles or more, 100 millimoles or more, 125 millimoles or more, 150 millimoles or more, 175 millimoles or more, 200 millimoles or more, 225 millimoles or more, 250 millimoles or more, 275 millimoles or more, 300 millimoles or more, etc. per liter of the total composition of the present disclosure. Furthermore, there is usually no upper limit to the amount of electrolyte in the present disclosure, as long as it can be present in solution as an electrolyte, relative to 1 L of the entire composition of the present disclosure; however, the amount may be, for example, 500 mmol or less, 475 mmol or less, 450 mmol or less, 425 mmol or less, 400 mmol or less, 375 mmol or less, 350 mmol or less, 325 mmol or less, 300 mmol or less, 275 mmol or less, 250 mmol or less, 225 mmol or less, 200 mmol or less, 175 mmol or less, 150 mmol or less, or the like.

[0067] In one embodiment, when the electrolyte of the present disclosure is an organic base or the like, it is typically present at 20 millimoles or more per liter of the total composition. In another embodiment, the electrolyte of the present disclosure can be, for example, 10 millimoles or more, 20 millimoles or more, 30 millimoles or more, 40 millimoles or more, 50 millimoles or more, 60 millimoles or more, 70 millimoles or more, 80 millimoles or more, 90 millimoles or more, 100 millimoles or more, 125 millimoles or more, 150 millimoles or more, 175 millimoles or more, 200 millimoles or more, etc. per liter of the total composition of the present disclosure. Furthermore, when the electrolyte of the present disclosure is an organic electrolyte, there is usually no upper limit as long as it can be present in solution as an electrolyte in 1 L of the entire composition of the present disclosure, but the upper limit may be, for example, 500 mmol or less, 475 mmol or less, 450 mmol or less, 425 mmol or less, 400 mmol or less, 375 mmol or less, 350 mmol or less, 325 mmol or less, 300 mmol or less, 275 mmol or less, 250 mmol or less, 225 mmol or less, 200 mmol or less, 175 mmol or less, 150 mmol or less, or the like.

[0068] In one embodiment, the electrolyte used in the present disclosure comprises an acid selected from acetic acid, citric acid, tartaric acid, glutamic acid, succinic acid, malic acid, and aspartic acid, or a salt thereof. Without wishing to be bound by theory, these electrolytes have been found to be particularly effective in preserving proteins.

[0069] In one embodiment, the electrolyte used in the present disclosure comprises an organic acid selected from acetic acid, succinic acid, and aspartic acid, or a salt thereof.

[0070] In one embodiment, the electrolyte used in the present disclosure includes acetic acid or a salt thereof. These electrolytes have been found to be particularly effective for preserving proteins. Furthermore, without wishing to be bound by theory, the technology of the present disclosure can be used to stably maintain the pH of solution formulations. Furthermore, acetic acid has a proven track record as a pharmaceutical additive and is highly safe.

[0071] In one embodiment, the electrolyte used in the present disclosure includes sodium acetate. These electrolytes have been found to be particularly effective for preserving proteins. Furthermore, without wishing to be bound by theory, the technology of the present disclosure can be used to stably maintain the pH of solution formulations. Furthermore, sodium acetate has a proven track record of use as a pharmaceutical additive and is highly safe.

[0072] In one embodiment, the electrolyte used in the present disclosure comprises at least one organic base selected from arginine, histidine, and tris(hydroxymethyl)aminomethane, or a salt thereof. These electrolytes have been found to be particularly effective for preserving proteins. Furthermore, without wishing to be bound by theory, the technology of the present disclosure can be used to stably maintain the pH of solution formulations. Furthermore, they have a proven track record of use as pharmaceutical additives and are highly safe. Preferably, the organic base is arginine. Without wishing to be bound by theory, these electrolytes have been found to be particularly effective for preserving proteins. Furthermore, they can stably maintain the pH of solution formulations. Furthermore, they have a proven track record of use as pharmaceutical additives and are highly safe.

[0073] In one embodiment, the electrolyte used in the present disclosure may be sodium chloride, potassium chloride, magnesium chloride, calcium chloride, sodium carbonate, sodium phosphate, potassium phosphate, or the like, and at least one or two or more of these may be used.

[0074] In one embodiment, the electrolyte used in the present disclosure comprises sodium chloride.

[0075] In one embodiment, the electrolyte used in the present disclosure includes cyclic oligosaccharides or derivatives thereof, such as substituted or unsubstituted (optionally substituted) α-CD, β-CD, and γ-CD, or derivatives thereof (e.g., hydroxypropyl, sulfobutyl ether, etc.). Preferably, the cyclic oligosaccharides or derivatives thereof used are hydroxypropyl-β-cyclodextrin (HP-β-CD) or sulfobutyl ether-β-cyclodextrin (SBE-β-CD).

[0076] In one embodiment, the cyclic oligosaccharides or derivatives thereof used in the present disclosure are typically present at 20 mmol or more per liter of the total composition of the present disclosure, and may be present at, for example, 10 mmol or more, 20 mmol or more, 30 mmol or more, 40 mmol or more, 50 mmol or more, 60 mmol or more, 70 mmol or more, 80 mmol or more, 90 mmol or more, 100 mmol or more, 125 mmol or more, 150 mmol or more, 175 mmol or more, 200 mmol or more, etc. per liter of the total composition of the present disclosure. Furthermore, when the electrolyte of the present disclosure is an organic electrolyte, there is usually no upper limit as long as it can be present in solution as an electrolyte in 1 L of the entire composition of the present disclosure, but the upper limit may be, for example, 500 mmol or less, 475 mmol or less, 450 mmol or less, 425 mmol or less, 400 mmol or less, 375 mmol or less, 350 mmol or less, 325 mmol or less, 300 mmol or less, 275 mmol or less, 250 mmol or less, 225 mmol or less, 200 mmol or less, 175 mmol or less, 150 mmol or less, or the like.

[0077] In one embodiment, the cyclic oligosaccharide or derivative thereof and the electrolyte used in the present disclosure are contained in equal molar amounts. In the present disclosure, "equal molar amounts" generally means a ratio of 1:2 to 2:1, preferably 1.5:1 to 1:1.5, 1.2:1 to 1:1.2, more preferably 1.1:1 to 1:1.1, and also preferably completely identical.

[0078] In one embodiment, the proteins used in the present disclosure may be antibodies, antigens, hormones, cytokines, enzymes, capsids, and the like.

[0079] Antibodies used in the present disclosure include the following. Antibodies used in the present disclosure include the following. For example, monoclonal antibodies and polyclonal antibodies against specific antigens, humanized antibodies, and chimeric antibodies are included. Fragment antibodies and reconfigurable single-chain antibodies (scFv) are also useful. Furthermore, it is desirable that these antibodies have high specificity and affinity for the target antigen. Antibodies can be widely applied in pharmaceuticals, diagnostics, and treatments, and antibodies with these properties play an important role in the present disclosure and are therefore encompassed within the scope of the present disclosure.

[0080] Antigens used in the present disclosure include the following. Examples include peptide antigens, protein antigens, carbohydrate antigens, and nucleic acid antigens. Cell surface antigens, viral antigens, and bacterial antigens are also useful. Furthermore, it is desirable that these antigens have the ability to induce a specific immune response. Antigens can be widely applied to vaccine development, diagnostic reagents, and therapeutic peptides, and antigens with these properties play an important role in the present disclosure and are therefore encompassed within the scope of the present disclosure.

[0081] Hormones that can be used in the present disclosure include the following. Examples include peptide hormones, steroid hormones, and amine hormones. Growth hormone, insulin, estrogen, and testosterone are also useful. Furthermore, it is desirable that these hormones have high specificity and effect on target cells. Hormones can be widely applied as endocrine therapy, metabolic regulation, and diagnostic tools, and hormones with these properties play an important role in the present disclosure and are therefore encompassed within the scope of the present disclosure.

[0082] Cytokines used in the present disclosure include interleukins, interferons, tumor necrosis factors, growth factors, and chemokines. Also included are interleukin-1 (IL-1), interleukin-2 (IL-2), interleukin-6 (IL-6), interleukin-10 (IL-10), interferon-α (IFN-α), interferon-β (IFN-β), interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), tumor necrosis factor-β (TNF-β), growth factor-β (TGF-β), erythropoietin (EPO), granulocyte colony-stimulating factor (G-CSF), macrophage colony-stimulating factor (MCSF), and macrophage colony-stimulating factor (MCF). Useful cytokines include CSF, platelet-derived growth factor (PDGF), fibroblast growth factor (FGF), nerve growth factor (NGF), hepatocyte growth factor (HGF), interleukin-8 (IL-8), interleukin-12 (IL-12), interleukin-15 (IL-15), interleukin-17 (IL-17), chemokines (CCL2, CCL3, CCL4), chemokines (CXCL8, CXCL10, CXCL12), tumor endothelial growth factor (VEGF), leptin, resistin, and adiponectin. These cytokines play important roles in regulating immune responses, promoting cell proliferation, and controlling inflammatory responses, and it is expected that their properties will be utilized in the present disclosure, and they are encompassed within the scope of the present disclosure.

[0083] Enzymes useful in the present disclosure include the following: oxidoreductases, hydrolases, transferases, isomerases, and synthetases. Specific examples include catalase, peroxidase, lipase, amylase, protease, kinase, phosphatase, dehydrogenase, transferase, isomerase, ligase, nuclease, DNA polymerase, RNA polymerase, peptidase, lactase, urease, lysozyme, cellulase, glucose oxidase, collagenase, hyaluronidase, trypsin, chymotrypsin, pepsin, renin, elastase, sucrase, and invertase. These enzymes promote specific chemical reactions, play important roles in regulating metabolic processes, and decomposition and synthesis of biomolecules. It is anticipated that these properties will be utilized in the present disclosure, and they are encompassed within the scope of the present disclosure.

[0084] Examples of capsids that can be used in the present disclosure include capsids derived from various viruses. For example, adenovirus, adeno-associated virus (AAV), retrovirus, lentivirus, herpesvirus, parvovirus, papillomavirus, influenza virus, coronavirus, poliovirus, Ebola virus, sarcoma virus, norovirus, rotavirus, Sendai virus, myelocytovirus, calicivirus, vaccinia virus, bocavirus, cytomegalovirus, and Newcastle disease virus are useful. These capsids play an important role in gene transfer, gene therapy products, oncolytic viruses, vaccine development, and the construction of viral vector systems, and their properties are expected to be utilized in the present disclosure, and are encompassed within the scope of the present disclosure.

[0085] In one embodiment, when the present disclosure uses capsids, the particle concentration of the capsids is typically 1×10 １３ In another embodiment, the particle concentration is 1 x 10 capsid particles / mL or more. １２ Capsid particles / mL or more, 5 x 10 １２ Capsid particles / mL or more 1 x 10 １３Capsid particles / mL or more, 5 x 10 １３ Capsid particles / mL or more, 1 x 10 １４ Capsid particles / mL or more, 5 x 10 １４ Capsid particles / mL or more, 1 x 10 １５ The concentration may be greater than or equal to 1000 capsid particles / mL.

[0086] In another aspect, the present disclosure provides a composition of the present disclosure as a pharmaceutical composition. The pharmaceutical composition may be a drug substance or a final formulation.

[0087] (Preservation of AAV) The present disclosure provides a technique for preserving adeno-associated virus (AAV). In one aspect, the present disclosure provides a composition for preserving adeno-associated virus (AAV) capsids, comprising a cyclic oligosaccharide or a derivative thereof. The provision of such a preservative is extremely useful for preserving AAV capsids.

[0088] In another aspect, the present disclosure provides a composition comprising an AAV capsid and an organic electrolyte, or a derivative thereof and / or an ion thereof, which is used to preserve the AAV capsid because such a composition has a higher preservation effect on the AAV capsid than when the other components are not included. One notable aspect of the present disclosure is that a composition comprising an organic electrolyte and / or an ion thereof has been found to be extremely effective in preserving AAV capsids.

[0089] In another aspect, the present disclosure provides a composition comprising an AAV capsid and a cyclic oligosaccharide or a derivative thereof.

[0090] In one embodiment, the organic electrolyte used in the present disclosure comprises at least one organic base and at least one organic acid, and such organic electrolytes are particularly effective in preserving AAV capsids.

[0091] In one embodiment, the organic electrolyte used in the present disclosure comprises at least one organic base selected from arginine, histidine, and tris(hydroxymethyl)aminomethane.

[0092] In another embodiment, the organic electrolyte used in the present disclosure comprises at least one organic acid selected from acetic acid, citric acid, tartaric acid, glutamic acid, succinic acid, malic acid, and aspartic acid. It is understood that combinations of these specific organic electrolytes and organic bases are preferably used. In a particularly preferred embodiment, the organic electrolyte comprises at least one organic base selected from arginine, histidine, and tris(hydroxymethyl)aminomethane, and at least one organic acid selected from acetic acid, citric acid, tartaric acid, glutamic acid, succinic acid, malic acid, and aspartic acid. More preferably, in the composition of the present disclosure, the organic electrolyte comprises an organic base that is arginine and an organic acid selected from acetic acid, succinic acid, and aspartic acid.

[0093] In one embodiment, the organic electrolyte of the present disclosure comprises an organic base and is typically present at 20 mmol or more per liter of the total composition. The organic electrolyte can be, for example, 10 mmol or more, 20 mmol or more, 30 mmol or more, 40 mmol or more, 50 mmol or more, 60 mmol or more, 70 mmol or more, 80 mmol or more, 90 mmol or more, 100 mmol or more, 125 mmol or more, 150 mmol or more, 175 mmol or more, 200 mmol or more, etc. per liter of the total composition of the present disclosure. Furthermore, there is usually no upper limit to the amount per liter of the entire composition of the present disclosure as long as it can be present in solution as an organic electrolyte, but the amount may be, for example, 500 mmol or less, 475 mmol or less, 450 mmol or less, 425 mmol or less, 400 mmol or less, 375 mmol or less, 350 mmol or less, 325 mmol or less, 300 mmol or less, 275 mmol or less, 250 mmol or less, 225 mmol or less, 200 mmol or less, 175 mmol or less, 150 mmol or less, or the like.

[0094] When the organic electrolyte of the present disclosure comprises an organic acid, it is typically present at 100 millimoles or more per liter of the total composition of the present disclosure. In certain embodiments, the electrolyte of the present disclosure can be, for example, 10 millimoles or more, 20 millimoles or more, 30 millimoles or more, 40 millimoles or more, 50 millimoles or more, 60 millimoles or more, 70 millimoles or more, 80 millimoles or more, 90 millimoles or more, 100 millimoles or more, 125 millimoles or more, 150 millimoles or more, 175 millimoles or more, 200 millimoles or more, 225 millimoles or more, 250 millimoles or more, 275 millimoles or more, 300 millimoles or more, etc. per liter of the total composition of the present disclosure. Furthermore, there is usually no upper limit to the amount of electrolyte in the present disclosure, as long as it can be present in solution as an electrolyte, relative to 1 L of the entire composition of the present disclosure; however, the amount may be, for example, 500 mmol or less, 475 mmol or less, 450 mmol or less, 425 mmol or less, 400 mmol or less, 375 mmol or less, 350 mmol or less, 325 mmol or less, 300 mmol or less, 275 mmol or less, 250 mmol or less, 225 mmol or less, 200 mmol or less, 175 mmol or less, 150 mmol or less, or the like.

[0095] The present disclosure further includes, for example, cyclic oligosaccharides, one or more divalent inorganic salts or carbohydrates.

[0096] In an exemplary embodiment, the divalent inorganic salt is selected from magnesium chloride and calcium chloride. Without wishing to be bound by theory, it is believed that the use of a divalent inorganic salt or a sugar or the like enhances the electric field effect when in solution, thereby enhancing the preservation effect.

[0097] Carbohydrates used in the present disclosure can be glucose, sucrose, trehalose, mannitol, etc. Without wishing to be bound by theory, it is believed that these carbohydrates are effective because they interact with the protein surface and stabilize the higher-order structure.

[0098] In one embodiment, the particle concentration of AAV capsids is 1×10 １３ capsid particles / mL or more. Alternatively, the capsid particle concentration is typically 1 x 10 １３In another embodiment, the particle concentration is 1 x 10 capsid particles / mL or more. １２ Capsid particles / mL or more, 5 x 10 １２ Capsid particles / mL or more 1 x 10 １３ Capsid particles / mL or more, 5 x 10 １３ Capsid particles / mL or more, 1 x 10 １４ Capsid particles / mL or more, 5 x 10 １４ capsid parti cles / mL or more, 1×10 １５ The concentration may be greater than or equal to 1000 capsid particles / mL.

[0099] In one embodiment, the compositions of the present disclosure may further comprise one or more surfactants.

[0100] In one embodiment, the AAV capsid of the present disclosure is an adeno-associated virus vector, i.e., it is understood that the preservation effect is also present even when the AAV capsid is used in the form of an adeno-associated virus vector.

[0101] In one embodiment, vector genome (vg) / mL can be used as a unit of particle concentration of AAV capsids in adeno-associated viral vectors. This unit refers to the concentration of nucleic acid encapsulated in AAV capsids, and in a sufficiently purified adeno-associated viral vector preparation, vector genome / mL and capsid particles / mL can be treated as the same value. When an adeno-associated viral vector preparation contains a large amount of AAV capsids that do not encapsulate nucleic acid (called empty capsids), the vector genome / mL is corrected for the rate of empty capsid contamination and converted to capsid particles / mL.

[0102] Formulations In an exemplary aspect, the compositions of the present disclosure may be provided as a formulation.

[0103] In some embodiments, in the formulation, the protein is present in the formulation at a concentration of at least 0.01 mg / mL. In one example, the protein is present in the formulation at a concentration of at least 0.05 mg / mL. In one example, the protein is present in the formulation at a concentration of at least 0.1 mg / mL. In one example, the protein is present in the formulation at a concentration of at least 0.5 mg / mL. In one example, the protein is present in the formulation at a concentration of at least 1 mg / mL. In one example, the protein is present in the formulation at a concentration of at least 2 mg / mL. In one example, the protein is present in the formulation at a concentration of at least 5 mg / mL. In one example, the protein is present in the formulation at a concentration of at least 10 mg / mL. In one example, the protein is present in the formulation at a concentration of at least 20 mg / mL. In one example, the protein is present in the formulation at a concentration of at least 30 mg / mL. In one example, the protein is present in the formulation at a concentration of at least 40 mg / mL. In one example, the protein is present in the formulation at a concentration of at least 50 mg / mL. In one example, the protein is present in the formulation at a concentration of at least 60 mg / mL. In one example, the protein is present in the formulation at a concentration of at least 70 mg / mL. In one example, the protein is present in the formulation at a concentration of at least 80 mg / mL. In one example, the protein is present in the formulation at a concentration of at least 90 mg / mL. In one example, the protein is present in the formulation at a concentration of at least 100 mg / mL. In one example, the protein is present in the formulation at a concentration of at least 110 mg / mL. In one example, the protein is present in the formulation at a concentration of at least 120 mg / mL. In one example, the protein is present in the formulation at a concentration of at least 150 mg / mL. In one example, the protein is present in the formulation at a concentration of at least 200 mg / mL. In one example, the protein is present in the formulation at a concentration of at least 250 mg / mL. In one example, the protein is present in the formulation at a concentration of at least 300 mg / mL.

[0104] The formulations herein may also contain two or more proteins necessary for the particular indication being treated, preferably those with complementary activities that do not adversely affect other proteins. For example, it may be desirable to provide two or more antibodies that bind to a desired target (e.g., a receptor or antigen) in a single formulation. Such proteins are preferably present in combination in amounts effective for the intended purpose.

[0105] The compositions of the present disclosure may contain other additive substances as needed and may be pharmaceutically acceptable.

[0106] Excipients include, for example, sugars such as monosaccharides, disaccharides, cyclodextrins and polysaccharides, metallic salts, citric acid, tartaric acid, glycine, polyethylene glycol, Pluronic®, kaolin, silicic acid, or combinations thereof.

[0107] Examples of binders include starch paste using vegetable starch, pectin, xanthan gum, simple syrup, glucose solution, gelatin, tragacanth, methylcellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose, shellac, paraffin, polyvinylpyrrolidone, and combinations thereof.

[0108] Disintegrants include, for example, the above-mentioned starches, lactose, carboxymethyl starch, cross-linked polyvinylpyrrolidone, agar, laminaran powder, sodium bicarbonate, calcium carbonate, alginic acid or sodium alginate, polyoxyethylene sorbitan fatty acid esters, sodium lauryl sulfate, stearic acid monoglyceride, or salts thereof.

[0109] Examples of emulsifiers include sorbitan fatty acid esters, glycerin fatty acid esters, sucrose fatty acid esters, and propylene glycol fatty acid esters.

[0110] Examples of flow regulators and lubricants include silicates, talc, stearates or polyethylene glycol.

[0111] In addition to the above, if necessary, the composition may appropriately contain solubilizers, suspending agents, diluents, dispersants, surfactants, soothing agents, stabilizers, absorption promoters, bulking agents, moisturizing agents, humectants, humectants, adsorbents, flavoring agents, disintegration inhibitors, coating agents, colorants, preservatives, antiseptics, antioxidants, buffers, pH adjusters, isotonic agents, and the like that are commonly used in pharmaceutical compositions, cell preparations, and the like.

[0112] The additive substances are used to avoid or suppress the decomposition of the active ingredient by enzymes and the like in the subject's body, as well as to facilitate formulation and administration methods and maintain the dosage form and medicinal efficacy, and may be used appropriately as needed.

[0113] The dosage form of the formulation of the present disclosure is a liquid. Liquid formulations include any formulations that have fluidity. Specific examples include creams, ointments, gels, injections, suspensions, etc. Specific volumes and the like are not particularly limited as long as they are within the range known in the art for each dosage form. The formulation of the present disclosure may be manufactured according to conventional methods in the art.

[0114] The present disclosure provides a formulation in which the composition of the present disclosure is filled in a vial or syringe.

[0115] In some specific embodiments, the present disclosure relates to a container containing the formulation of the present disclosure. For example, the formulation may be packaged in a single-use or multi-use vial or a kit for single-dose administration units. In another embodiment of the present disclosure, an article of manufacture is provided that includes a container containing the formulation and may also provide instructions for its use. Suitable containers include, for example, bottles, vials (e.g., dual-chamber vials), syringes (such as single- or dual-chamber syringes), and test tubes. The container may be formed from a variety of materials, such as glass or plastic. Such containers or kits include both single- and multi-chamber pre-filled syringes. Exemplary pre-filled syringes are available from Vetter GmbH of Ravensburg, Germany, and Thermo Fisher Scientific Inc. of Lombardy, Italy. A label on or associated with the container holding the formulation may indicate instructions for reconstitution and / or use. The label may further indicate that the formulation is useful or intended for subcutaneous administration. The container holding the formulation may be a multi-use vial allowing for repeated administration (e.g., 2-6 administrations). The article of manufacture may further include a second container containing, for example, a suitable diluent (e.g., BWFI) for reconstitution of the lyophilized formulation. The article of manufacture may further include other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, needles, syringes, and package inserts with instructions for use.

[0116] The formulations of the present disclosure preferably contain appropriate buffers, preservatives, and stabilizers to enhance shelf life and stability. Examples of useful agents include phosphate buffer, glycerol, mannitol, trehalose, and polysorbate 80. The compositions are manufactured under sterile conditions and filtered through a sterile filter. They are then dispensed into vials or syringes in a sterile environment and sealed. These formulations are recommended for refrigeration, but can also be stored at room temperature. For example, in the case of insulin formulations, insulin drug substance is dissolved in phosphate buffer and glycerol, and trehalose and polysorbate 80 are added and mixed. The mixture is then filtered through a sterile filter and dispensed into sterile vials or syringes. The final product undergoes quality inspection before shipping. The formulations of the present disclosure have high stability and efficacy, making them suitable for use in clinical settings.

[0117] It is important that the materials of the vials and syringes used in this disclosure are carefully selected to ensure the stability and safety of the formulation. Glass and certain plastics are commonly used as vial materials. Glass vials are primarily made of borosilicate glass, characterized by their high chemical resistance and inertness. Plastic vials, on the other hand, are made of materials such as polypropylene (PP), polyethylene (PE), cyclic olefin copolymer (COC), and cycloolefin polymer (COP), which have the advantages of being lightweight and shatter-resistant. Plastic vials are often used as containers for gene therapy products.

[0118] Both glass and plastic syringes are used as syringe materials. Glass syringes have low reactivity with chemicals and are suitable for long-term storage. Plastic syringes are made from materials such as polypropylene (PP), polyethylene (PE), polycarbonate (PC), cyclic olefin copolymer (COC), and cycloolefin polymer (COP), and have the advantage of being lightweight and shatter-resistant. Plastic syringes are often used as containers for gene therapy products.

[0119] Furthermore, the syringe plunger is made of silicone-coated rubber or thermoplastic elastomer (TPE) to ensure smooth operation and leakage prevention. Furthermore, the rubber stoppers and seals of the vials and syringes are made of materials that are compatible with the drug solution and have low gas permeability, such as Viton (fluororubber) or chlorobutyl rubber. The use of these materials allows the formulations disclosed herein to maintain high quality and safety, enabling reliable use in medical settings.

[0120] The present disclosure provides kits that include a composition of the present disclosure (typically a pharmaceutical composition or a protein preservative).

[0121] The present disclosure provides a kit containing a composition of the present disclosure (typically a pharmaceutical composition or a protein preservative). The kit is designed to allow users to easily handle, store, and administer the composition. Specifically, the kit includes the following elements: 1) a sterile vial or syringe filled with the composition, 2) an intravenous bag and a sterile needle or cannula for administration, 3) instructions for use, and 4) appropriate packaging for storage and transportation. This ensures accurate and safe use in clinical settings.

[0122] The present disclosure provides a kit containing a composition of the present disclosure (typically a pharmaceutical composition or a protein preservative). The kit is designed to improve the preservation and stability of protein pharmaceuticals, allowing users to easily perform preservation procedures. Typically, the kit of the present disclosure contains the components constituting the composition described in any one of claims 1 to 44, separated into compartments. Components that may be provided as a mixture of two types may be provided as a mixture. Specifically, the kit includes the following elements: 1) a sterile vial or ampoule filled with the protein preservative, 2) a sterile mixing tool (e.g., a syringe or pipette) for preservation procedures, 3) instructions for use, and 4) appropriate packaging for storage and transportation. This allows for accurate and safe use in research facilities and pharmaceutical companies. The kit of the present disclosure may also include instructions describing how to use and / or prepare the composition.

[0123] The kit includes the composition of the present disclosure as well as various additives to further enhance protein stability, such as sugars (trehalose, sucrose), amino acids (glycine, arginine), polyhydric alcohols (glycerol, mannitol), and polymers (polyethylene glycol).

[0124] The manufacturing method using the kit of the present disclosure can be exemplified as follows. For example, first, the components of the preservative are mixed under sterile conditions to prepare an appropriate solution. Next, this preservative solution is filtered through a sterile filter and dispensed into sterile vials or ampoules. After dispensing, the vials or ampoules are individually sealed and enclosed in a sterile package. Furthermore, sterilized mixing tools for the preservation process are also packaged under sterile conditions. Finally, these components are combined into a kit, which is then inspected for quality and shipped.

[0125] This kit significantly improves the shelf life and stability of protein pharmaceuticals, making it an important tool in research and development and manufacturing processes, thereby extending product expiration dates and ensuring product quality during storage.

[0126] The present disclosure provides a method for producing a composition of the present disclosure, the method comprising: 1) providing the protein; and 2) adding the electrolyte and / or the cyclic oligosaccharide or derivative thereof to the protein and mixing them. Such a method includes the following specific steps. Regarding 3) providing the protein, the following example is provided: First, the protein to be preserved is purified under appropriate conditions. This protein may be of various types, such as pharmaceuticals, enzymes, or antibodies. After purification, the protein is dissolved in an appropriate buffer (e.g., phosphate buffer, Tris buffer), and its concentration is adjusted to a certain level. The purity and concentration of the protein are confirmed by standard biochemical methods (e.g., SDS-PAGE, ELISA, Bradford method) or physicochemical methods (e.g., HPLC, capillary electrophoresis).

[0127] Next, a step of adding and mixing the electrolyte and / or the cyclic oligosaccharide or derivative thereof of the present disclosure to the protein, either separately or together, is carried out. Here, a storage stabilizer X is added to the protein solution. The electrolyte and / or the cyclic oligosaccharide or derivative thereof are as described elsewhere in this disclosure. These stabilizers prevent protein denaturation and preserve its biological activity. The amount of storage stabilizer X added needs to be optimized depending on the type and concentration of the protein.

[0128] As a specific example, when preparing a storage composition for insulin, electrolytes and / or the cyclic oligosaccharide or derivative thereof (final concentration: 0.5 M) and polysorbate 20 (final concentration: 0.02%) are added to an insulin solution and gently stirred to mix. This mixture is then filtered through a sterile filter and dispensed into sterile vials. The vials are sealed and stored under frozen or refrigerated conditions. The storage composition prepared according to the present disclosure enables long-term stable storage of proteins, significantly improving their usefulness as pharmaceuticals and research samples.

[0129] In another aspect, the present disclosure provides a method for suppressing protein aggregation using a composition of the present disclosure, the method comprising: 1) providing the protein; and 2) placing the electrolyte or an ion thereof and / or a cyclic oligosaccharide or a derivative thereof under conditions in which the electrolyte or an ion thereof and / or a cyclic oligosaccharide or a derivative thereof is brought into contact with the protein.

[0130] Such a method includes the following specific steps. First, in the step of providing a protein, the protein for which aggregation formation is to be suppressed is purified under appropriate conditions so that it can be provided in a preservative method. This protein can be a wide variety of proteins, including pharmaceuticals, enzymes, and antibodies. After purification, the protein is dissolved in an appropriate buffer (e.g., phosphate buffer or Tris buffer) and its concentration is adjusted to a certain level. The purity and concentration of the protein are confirmed using standard biochemical methods (e.g., SDS-PAGE, ELISA, Bradford method) or physicochemical methods (e.g., HPLC, capillary electrophoresis).

[0131] Next, the step of placing electrolytes and / or cyclic oligosaccharides or derivatives thereof under conditions in which they can contact the protein can be carried out as follows: Electrolytes and / or cyclic oligosaccharides or derivatives thereof are added to the protein solution to suppress the formation of aggregates. The electrolytes and / or cyclic oligosaccharides or derivatives thereof are components described in the present disclosure, and such data can prevent protein aggregation and stabilize its structure. These stabilizers inhibit interactions between protein molecules and suppress the formation of aggregates. The amount of electrolytes and / or cyclic oligosaccharides or derivatives thereof added is optimized depending on the type and concentration of the protein.

[0132] For example, to suppress the formation of aggregates in a monoclonal antibody, electrolytes and / or cyclic oligosaccharides or derivatives thereof (final concentration: 0.5 M) and polysorbate 80 (final concentration: 0.02%) are added to an antibody solution and gently stirred to mix. The mixture is then filtered through a sterile filter and dispensed into sterile vials. The vials are then sealed and stored under refrigerated conditions.

[0133] The composition produced by the method of the present disclosure effectively inhibits protein aggregation and maintains its biological activity during long-term storage. This improves the quality of pharmaceuticals and research samples and increases their reliability during use. Furthermore, this method is suitable for industrial applications due to its simple manufacturing process and ease of scale-up.

[0134] In one embodiment, the present disclosure provides a method for enriching a protein contained in a composition using a composition of the present disclosure, the method comprising the steps of: 1) providing the protein; and 2) placing the electrolyte or ions thereof and / or cyclic oligosaccharides or derivatives thereof under conditions that allow the electrolyte or ions thereof and / or cyclic oligosaccharides or derivatives thereof to contact the protein. Such a method includes the following specific steps:

[0135] The step of first providing a protein can be carried out as follows: First, the protein to be concentrated is purified under appropriate conditions. This protein can be a wide variety of proteins, including pharmaceuticals, enzymes, and antibodies. After purification, the protein is dissolved in an appropriate buffer (e.g., phosphate buffer or Tris buffer), and its initial concentration is adjusted to a certain value. The purity and concentration of the protein are confirmed using standard biochemical techniques (e.g., SDS-PAGE, ELISA, Bradford method) or physicochemical techniques (e.g., HPLC, capillary electrophoresis).

[0136] Next, the step of placing the electrolyte or its ions and / or cyclic oligosaccharides or derivatives thereof under conditions in which they contact the protein can be carried out as follows. For example, next, electrolytes or their ions and / or cyclic oligosaccharides or derivatives thereof are added to the protein solution to achieve a high protein concentration. The electrolytes or their ions and / or cyclic oligosaccharides or derivatives thereof can improve the solubility of the protein and prevent aggregation, resulting in a high protein concentration. These additives play a role in adjusting the interactions between protein molecules and maintaining a stable solution even at high concentrations. The amount of electrolytes or their ions and / or cyclic oligosaccharides or derivatives added needs to be optimized depending on the type of protein and the desired concentration.

[0137] For example, to increase the concentration of a monoclonal antibody, arginine (final concentration 0.5 M) and cyclodextrin (final concentration 5%) are added to the antibody solution and gently mixed by stirring. This mixture is then concentrated to the target concentration using an ultrafiltration or concentrator. The concentrated antibody solution is filtered through a sterile filter and dispensed into sterile vials. The vials are then sealed and stored under refrigerated conditions.

[0138] For example, to increase the concentration of AAV capsids, electrolytes and / or cyclic oligosaccharides or derivatives thereof (final concentration: 0.5 M) and poloxamer 188 (final concentration: 0.01%) are added to the capsid solution and gently stirred to mix. This mixture is then concentrated to the target concentration using an ultrafiltration or concentrator. The concentrated capsid solution is filtered through a sterile filter and dispensed into sterile vials. The vials are sealed and stored under refrigerated or frozen conditions.

[0139] The highly concentrated protein composition produced by the method of the present disclosure is suitable for use as an active ingredient in pharmaceuticals, and exhibits high efficacy in small amounts. Furthermore, the method is suitable for mass production due to its simple manufacturing process and ease of scale-up. This improves the efficiency and economic viability of the product, and is expected to play an important role in the medical and research fields.

[0140] Therapeutic Treatment Using the Formulations of the Present Disclosure: In the present disclosure, "treatment" refers to both therapeutic treatment and prophylactic or preventative measures. Those in need of treatment include those who already have the disorder and those in whom the disorder is to be prevented. Treatment includes alleviating the symptoms of the disorder, improving the quality of life of the subject, and any relief or improvement of the subject, such as stabilizing the disorder, preventing the disorder from worsening, curing, or reducing the risk of recurrence.

[0141] In this disclosure, "subject" and "patient" are used interchangeably and generally refer to a mammal receiving treatment. For purposes of treatment, "mammal" refers to any animal classified as a mammal, e.g., humans, domestic and farm animals, and zoo, sport, or pet animals, such as dogs, horses, rabbits, cows, pigs, hamsters, gerbils, mice, ferrets, rats, cats, etc. In some embodiments, the subject is a human.

[0142] As used herein, a "disorder" is any condition that would benefit from treatment with a protein. This includes chronic and acute disorders or diseases, including pathological conditions that predispose a mammal to the disorder in question. Non-limiting examples of disorders treated herein include carcinoma and inflammation.

[0143] In the present disclosure, a "therapeutically effective amount" is at least the minimum concentration necessary to produce a measurable treatment of a particular disorder. Therapeutically effective amounts of known protein drugs are well known in the art, but effective amounts of the proteins set forth below can be determined by standard techniques within the skill of a person skilled in the art, e.g., an ordinary physician.

[0144] Proteins covered by this disclosure may be formulated according to the present disclosure in either liquid or lyophilized form. Routes of administration follow known and accepted methods of injection or infusion, either as single or multiple boluses or over time in a suitable manner, for example, subcutaneous, intravenous, intraperitoneal, intramuscular, intraarterial, intralesional, or intraarticular, by topical administration, inhalation, or by slow or sustained release means.

[0145] For the treatment of disorders, the appropriate dosage of the active agent will depend on the type of disorder being treated, the severity and course of the disorder, whether the agent is being administered for prophylactic or therapeutic purposes, previous therapy, the patient's medical history and response to the agent, and the discretion of the treating physician, as defined above. The agent is preferably administered to the patient at one time or over a series of treatments.

[0146] The formulations of the present disclosure, including but not limited to non-lyophilized liquid formulations and reconstituted formulations, can be administered to a mammal, e.g., a human, in need of treatment with a protein according to known methods, such as intravenous administration as a bolus or continuous infusion over a period of time, intramuscular, intraperitoneal, intracerebrospinal, subcutaneous, intra-articular, intrasynovial, intrathecal, oral, topical, or inhalation routes. In some embodiments, the formulation is administered to the mammal by subcutaneous (i.e., under the skin) administration. For such purposes, the formulation can be injected using a syringe. However, other devices are available for administering the formulation, such as injection devices (e.g., Inject-ease® and Genject® devices); pen-type injectors (such as GenPen®); auto-injector devices, needle-free devices (e.g., MediJector® and BioJector®); and subcutaneous patch delivery systems.

[0147] The appropriate dosage of the protein (a "therapeutically effective amount") will depend, for example, on the condition being treated, the severity and course of the condition, whether the protein is administered for prophylactic or therapeutic purposes, previous therapy, the patient's medical history and response to the protein, the type of protein used, and the discretion of the attending physician. The protein is suitably administered to the patient at one time or over a series of treatments, and can be administered to the patient at any time from the time of diagnosis. The protein can be administered as the sole treatment or in conjunction with other drugs or therapies useful in treating the condition in question.

[0148] The present disclosure has been described above by showing preferred embodiments for ease of understanding. The present disclosure will be described below based on examples. However, the above description and the following examples are provided for illustrative purposes only and are not intended to limit the present disclosure. Therefore, the scope of the present disclosure is not limited to the embodiments or examples specifically described herein, but is limited only by the scope of the claims.

[0149] In this example, various functions of T cells with enhanced capacity were investigated. The specific reagents used were those listed in the examples, but equivalent products from other manufacturers (Sigma-Aldrich, Wako Pure Chemical Industries, Nakarai, R&D Systems, USCN Life Science INC, etc.) can also be used. Animal experiments were conducted in accordance with the university's experimental guidelines, based on the spirit of animal welfare.

[0150] The abbreviations used herein are as follows:

[0151]

[0152] The model proteins used were AAV6-CMV-EGFP, AAV2-CMV-EGFP, AAV8-CMV-EGFP, and AAV9-CMV-EGFP (gene therapy products), IgG (antibody drug), and insulin (non-antibody protein).

[0153] (Comparative Example: Formulation of Commercially Available AAV Vector Products) Commercially available AAV vector products (for example, Zolgensma and Luxturna, sold in Japan by Novartis Pharma) widely use high concentrations of sodium chloride and surfactants for stabilization, and no formulations containing organic acids or cyclodextrins have been confirmed. Zolgensma package insert: https: / / www.drs-net.novartis.co.jp / siteassets / comMon / pdf / zol / pi / pi_zol_202302.pdf Luxturna package insert: https: / / www.drs-net.novartis.co.jp / siteassets / comMon / pdf / zol / pi / pi_zol_202302.pdf jp / siteassets / comMon / pdf / lux / pi / pi_lux_202306. pdf Package Insert - HEMGENIX: https: / / www. fda. gov / media / 163467 / download

[0154]

[0155] The structures of HP-α-CD, HP-β-CD and HP-γ-CD are as follows:

[0156]

[0157] [Example 1] Protein aggregation onset temperature (T ａｇｇ ) and protein denaturation temperature (T ｍ Using the protein property evaluation device UNcle manufactured by Unchained Labs, a solution (8.8 μL) containing the target protein prepared under each solvent condition was heated from 25°C to 95°C at a heating rate of 1°C / min, and the temperature dependence of the scattered light intensity at 266 nm was evaluated. As a representative diagram, the temperature dependence of the scattered light intensity for a sample solution of 1.5 mg / mL IgG, 5 mM Cit / NaOH, pH 6.0 is shown in Figure 1-1. The IgG used was human plasma-derived IgG from Golden West Diagnostics. The sample solution was filtered through a 0.22 μm hydrophilic PVDF membrane before use in the measurement. The protein aggregation onset temperature (T ａｇｇ ) was calculated and used as an index of protein aggregation.

[0158] Using the protein physical property evaluation device UNcle from Unchained Labs, a solution (8.8 μL) containing the target protein prepared under each solvent condition was heated from 25 °C to 95 °C at a heating rate of 1 °C / min, and the λ value was measured, which is the wavelength that gives the center of gravity of the internal fluorescence spectrum (300-430 nm) when excited at 266 nm. ＢＣＭ The temperature dependence of the thickness (nm) was evaluated. １３ λ in a sample solution of vg / mL AAV6-CMV-EGFP, 10 mM His / HCl, 190 mM NaCl, 0.33% (w / v) PX188, pH 6.0 ＢＣＭ The temperature dependence of λ is shown in Figure 1-2. AAV6-CMV-EGFP was manufactured and purified by Synplogen. The sample solution was filtered through a 0.22 μm hydrophilic PVDF membrane before use in the measurement. λ ＢＣＭ The protein denaturation temperature (T ｍ ) was calculated using UNcle Analysis software (V5.03) and used as an index of the stability of the protein's three-dimensional structure, which is known to have a significant effect on protein aggregation.

[0159] Table 1 shows an overview of Examples 1 and onwards. Table 2 shows the sources (manufacturers) of the reagents used in the following Examples. Table 3 shows a list of the stock solutions used in Examples 2 to 9.

[0160]

[0161]

[0162]

[0163] [Example 2] T in AAV6-CMV-EGFP by simultaneous addition of various electrolytes containing Cl and HP-β-CD ａｇｇ Solutions containing AAV6-CMV-EGFP were prepared under each solvent condition, and the T ａｇｇ was calculated. AAV6-CMV-EGFP drug substance A (6.3 × 10 １２vg / mL AAV6-CMV-EGFP, 8.1mM Na ２ HPO ４ , 1.5 mM KH ２ P.O. ４ , 137.9 mM NaCl, 2.7 mM KCl, 0.005% (w / v) PX188, pH 7.2) manufactured and purified by Synplogen. 3.1 μL of each of the preparation solutions F, E1, E2, E3, E4, C1, EC1, EC2, EC3, and EC4 listed in Table 3 was added to 6.2 μL of the AAV6-CMV-EGFP drug substance to prepare a sample with no additional electrolytes or HP-β-CD (Free), a sample with 100 mM NaCl (100 mM NaCl), a sample with 100 mM KCl (100 mM KCl), and a sample with 100 mM MgCl. ２ Samples supplemented with (100 mM MgCl ２ ), a sample with 100 mM Tris-HCl added (100 mM Tris-HCl), a sample with 100 mM HP-β-CD added (100 mM HP-β-CD), a sample with 100 mM NaCl and 100 mM HP-β-CD added (100 mM NaCl + 100 mM HP-β-CD), a sample with 100 mM KCl and 100 mM HP-β-CD added (100 mM KCl + 100 mM HP-β-CD), and a sample with 100 mM MgCl ２ and a sample containing 100 mM HP-β-CD (100 mM MgCl ２ A total of 10 samples were prepared: one containing 100 mM Tris-HCl (100 mM Tris-HCl + 100 mM HP-β-CD), and one containing 100 mM Tris-HCl and 100 mM HP-β-CD (100 mM Tris-HCl + 100 mM HP-β-CD).

[0164] Various electrolytes containing Cl (NaCl, KCl, MgCl ２ 2-1, 2-2, 2-3, and 2-4 show the temperature dependence of scattered light intensity when HP-β-CD and HP-β-CD were added alone and simultaneously. ａｇｇThe results are shown in Table 4. When any electrolyte was used, the T was significantly higher when the electrolyte and HP-β-CD were added simultaneously than when they were added separately. ａｇｇ Change in (ΔT ａｇｇ ) was confirmed to be the largest. ２ Regarding the above, by adding it simultaneously with HP-β-CD, protein aggregation was suppressed to the extent that no clear protein aggregation was detectable even at 95° C., which is the upper measurement temperature limit of the instrument.

[0165]

[0166] [Example 3] T in AAV6-CMV-EGFP by simultaneous addition of various electrolytes including Na and HP-β-CD ａｇｇ Solutions containing AAV6-CMV-EGFP were prepared under each solvent condition, and the T ａｇｇ was calculated. AAV6-CMV-EGFP drug substance B (1.8 × 10 １３ vg / mL AAV6-CMV-EGFP, 8.1mM Na ２ HPO ４ , 1.5 mM KH ２ P.O. ４, 137.9 mM NaCl, 2.7 mM KCl, 0.005% (w / v) PX188, pH 7.2) manufactured and purified by Synplogen Co., Ltd. was used. To 7 μL of the AAV6-CMV-EGFP drug substance, 3.5 μL of each of the preparation solutions F, E1, E5, E6, E7, E8, E9, E10, C1, EC1, EC5, EC6, EC7, EC8, EC9, and EC10 listed in Table 3 was added. This gave a sample with no additional electrolytes or HP-β-CD added (Free), a sample with 100 mM NaCl added (100 mM NaCl), a sample with 100 mM Na-Ace added (100 mM Na-Ace), a sample with 100 mM Na-Asp added (100 mM Na-Asp), a sample with 100 mM Na-Suc added (100 mM Na-Suc), a sample with 100 mM Na-Mal added (100 mM Na-Mal), and a sample with 100 mM Na-Mal added (100 mM Na-Mal). The samples were: Na-Tar added (100mM Na-Tar), 100mM Na-Cit added (100mM Na-Cit), 100mM HP-β-CD added (100mM HP-β-CD), 100mM NaCl and 100mM HP-β-CD added (100mM NaCl + 100mM HP-β-CD), 100mM Na-Ace and 100mM HP-β-CD added (100mM Na-Ace + 100mM HP-β-CD), 100mM Na-Asp and 100mM HP-β-CD added (100mM Na-Asp + 100mM HP-β-CD), 100mM Na-Suc and 100mM A total of 16 samples were prepared: a sample containing HP-β-CD (100 mM Na-Suc + 100 mM HP-β-CD), a sample containing 100 mM Na-Mal and 100 mM HP-β-CD (100 mM Na-Mal + 100 mM HP-β-CD), a sample containing 100 mM Na-Tar and 100 mM HP-β-CD (100 mM Na-Tar + 100 mM HP-β-CD), and a sample containing 100 mM Na-Cit and 100 mM HP-β-CD (100 mM Na-Cit + 100 mM HP-β-CD).

[0167] Figures 3-1, 3-2, 3-3, 3-4, 3-5, 3-6, and 3-7 show a comparison of the temperature dependence of scattered light intensity when various electrolytes containing Na (NaCl, Na-Ace, Na-Asp, Na-Suc, Na-Mal, Na-Tar, Na-Cit) and HP-β-CD are added alone or simultaneously. ａｇｇ The results are shown in Table 5. When any electrolyte was used, the T was significantly higher when the electrolyte and HP-β-CD were added simultaneously than when they were added separately. ａｇｇ Change in (ΔT ａｇｇ ) was confirmed to be the largest.

[0168]

[0169] [Example 4] T in AAV6-CMV-EGFP by simultaneous addition of various electrolytes containing Arg and HP-β-CD ａｇｇ Solutions containing AAV6-CMV-EGFP were prepared under each solvent condition, and the T ａｇｇ was calculated. AAV6-CMV-EGFP drug substance B (1.8 × 10 １３ vg / mL AAV6-CMV-EGFP, 8.1mM Na ２ HPO ４ , 1.5 mM KH ２ P.O. ４, 137.9 mM NaCl, 2.7 mM KCl, 0.005% (w / v) PX188, pH 7.2) manufactured and purified by Synplogen Co., Ltd. was used. To 7 μL of AAV6-CMV-EGFP drug substance, 3.5 μL of each of the preparation solutions F, E11, E12, E13, E14, C1, EC11, EC12, EC13, and EC14 listed in Table 3 was added. This gave a sample with no additional electrolytes or HP-β-CD (Free), a sample with 100 mM Arg-HCl (100 mM Arg-HCl), a sample with 100 mM Arg-Ace (100 mM Arg-Ace), a sample with 100 mM Arg-Suc (100 mM Arg-Suc), a sample with 100 mM Arg-Asp (100 mM Arg-Asp), and a sample with 100 mM HP-β-CD (100 mM A total of 10 samples were prepared: a sample containing 100 mM Arg-HCl and 100 mM HP-β-CD (100 mM Arg-HCl + 100 mM HP-β-CD), a sample containing 100 mM Arg-Ace and 100 mM HP-β-CD (100 mM Arg-Ace + 100 mM HP-β-CD), a sample containing 100 mM Arg-Suc and 100 mM HP-β-CD (100 mM Arg-Suc + 100 mM HP-β-CD), and a sample containing 100 mM Arg-Asp and 100 mM HP-β-CD (100 mM Arg-Asp + 100 mM HP-β-CD).

[0170] Figures 4-1, 4-2, 4-3, and 4-4 show a comparison of the temperature dependence of scattered light intensity when various electrolytes containing Arg (Arg-HCl, Arg-Ace, Arg-Suc, Arg-Asp) and HP-β-CD were added alone or simultaneously. ａｇｇ The results are shown in Table 6. When any electrolyte was used, the T was significantly higher when the electrolyte and HP-β-CD were added simultaneously than when they were added separately. ａｇｇ Change in (ΔT ａｇｇ ) was confirmed to be the largest.

[0171]

[0172] [Example 5] T by simultaneous addition of Arg-Asp and HP-β-CD in AAV6-CMV-EGFP ｍ Using a total of 10 samples containing AAV6-CMV-EGFP prepared in Example 4, the T ｍ was calculated.

[0173] When various electrolytes containing Arg (Arg-HCl, Arg-Ace, Arg-Suc, Arg-Asp) and HP-β-CD were added alone or simultaneously, λ ＢＣＭ The temperature dependence of the thickness (nm) is compared in Figures 5-1, 5-2, 5-3, and 5-4. ｍ The results are shown in Table 7. When Asp was used as a counter ion for Arg, T ｍ was found to increase the most.

[0174]

[0175] [Example 6] T by divalent cation, sucrose, in AAV6-CMV-EGFP ａｇｇ , T ｍ Solutions containing AAV6-CMV-EGFP were prepared under each solvent condition, and the T ａｇｇ and T ｍ The AAV6-CMV-EGFP used was produced and purified by Synplogen.

[0176] 2 x 10 １３ vg / mL AAV6-CMV-EGFP, 10 mM His / HCl, 190 mM NaCl, 0.33% (w / v) PX188, pH 6.0 as the base condition (190 mM NaCl), 190 mM NaCl, 190 mM Arg-Asp, 190 mM Arg-Asp + 20 mM MgCl ２ , 190mM Arg-Asp+20mM CaCl ２ , or 190 mM Arg-Asp + 100 mM sucrose. ＢＣＭThe temperature dependence of T is compared in Figures 6-1 and 6-2. ｍ and T ａｇｇ are shown in Table 8. NaCl is replaced with Arg-Asp, Arg-Asp + MgCl ２ , Arg-Asp+CaCl ２ , Arg-Asp+sucrose, ａｇｇ and T ｍ Increases in both were observed.

[0177]

[0178] [Example 7] T in AAV6-CMV-EGFP by simultaneous addition of Arg-HCl and SBE-β-CD ａｇｇ Solutions containing AAV6-CMV-EGFP were prepared under each solvent condition, and the T ａｇｇ was calculated. AAV6-CMV-EGFP drug substance B (1.8 × 10 １３ vg / mL AAV6-CMV-EGFP, 8.1mM Na ２ HPO ４ , 1.5 mM KH ２ P.O. ４ , 137.9 mM NaCl, 2.7 mM KCl, 0.005% (w / v) PX188, pH 7.2) manufactured and purified by Synplogen Co., Ltd. was used. To 6.2 μL of the AAV6-CMV-EGFP drug substance, 3.1 μL of each of the preparation solutions F, E11, C2, and EC15 shown in Table 3 was added to prepare four samples in total: a sample to which no electrolytes or HP-β-CD were added (Free), a sample to which 100 mM Arg-HCl was added (100 mM Arg-HCl), a sample to which 100 mM SBE-β-CD was added (100 mM SBE-β-CD), and a sample to which 100 mM Arg-HCl and 100 mM SBE-β-CD were added (100 mM Arg-HCl + 100 mM SBE-β-CD).

[0179] FIG. 7 shows a comparison of the temperature dependence of scattered light intensity when Arg-HCl and SBE-β-CD were added alone and when they were added simultaneously. ａｇｇThe results are shown in Table 9. When Arg-HCl and SBE-β-CD were added simultaneously, T ａｇｇ Change in (ΔT ａｇｇ ) was confirmed to be the largest.

[0180]

[0181] [Example 8] T in AAV6-CMV-EGFP by simultaneous addition of Arg-HCl and HP-β-CD (20 mM, 40 mM) ａｇｇ Solutions containing AAV6-CMV-EGFP were prepared under each solvent condition, and the T ａｇｇ was calculated. AAV6-CMV-EGFP drug substance C (1.7 × 10 １３ vg / mL AAV6-CMV-EGFP, 12.5mM Bis-Tris propane / HCl, 50mM glycine, 250mM NaCl, 1mM MgCl ２ , 0.05% (w / v) PX188, pH 7.9) manufactured and purified by Synplogen Co., Ltd. was used. To 7 μL of AAV6-CMV-EGFP drug substance, 3.5 μL of the preparation solutions F, E15, E16, C3, C4, EC16, and EC17 shown in Table 3 were added to prepare a sample with no additional Arg-HCl or HP-β-CD (Free), a sample with 20 mM Arg-HCl (20 mM Arg-HCl), a sample with 40 mM Arg-HCl (40 mM Arg-HCl), a sample with 20 mM HP-β-CD (20 mM HP-β-CD), a sample with 40 mM HP-β-CD (40 mM HP-β-CD), and a sample with 20 mM Arg-HCl and 20 mM HP-β-CD (20 mM Arg-HCl + 20 mM A total of seven samples were prepared: one containing 40 mM HP-β-CD, one containing 40 mM Arg-HCl and 40 mM HP-β-CD (40 mM Arg-HCl + 40 mM HP-β-CD).

[0182] Figures 8-1 and 8-2 show a comparison of the temperature dependence of scattered light intensity when Arg-HCl and HP-β-CD were added alone and when they were added simultaneously. ａｇｇThe results are shown in Tables 10-1 and 10-2. Under all concentration conditions, the T was significantly higher when Arg-HCl and HP-β-CD were added simultaneously than when they were added alone. ａｇｇ Change in (ΔT ａｇｇ ) was confirmed to be the largest.

[0183]

[0184]

[0185] [Example 9] T by simultaneous addition of Arg-HCl and HP-β-CD in AAV2-CMV-EGFP, AAV8-CMV-EGFP, and AAV9-CMV-EGFP ａｇｇ Solutions containing AAV2-CMV-EGFP, AAV8-CMV-EGFP, and AAV9-CMV-EGFP were prepared under each solvent condition, and the T ａｇｇ was calculated. AAV2-CMV-EGFP drug substance (1.9 × 10 １３ vg / mL AAV2-CMV-EGFP, 10.1mM Na ２ HPO ４ , 1.8 mM KH ２ P.O. ４ , 336.8mM NaCl, 2.7mM KCl, 0.001% (w / v) PX188, pH 7.4 (Cat No. AAV2SP (VB010000-9394npt)-C)), AAV8-CMV-EGFP drug substance (1.8 x 10 １３ vg / mL AAV8-CMV-EGFP, 10.1mM Na ２ HPO ４ , 1.8 mM KH ２ P.O. ４ , 336.8mM NaCl, 2.7mM KCl, 0.001% (w / v) PX188, pH 7.4 (Cat No. AAV8SP (VB010000-9394npt)-C)), AAV9-CMV-EGFP drug substance (1.5 x 10 １３ vg / mL AAV9-CMV-EGFP, 10.1mM Na ２ HPO ４ , 1.8 mM KH ２ P.O. ４, 336.8 mM NaCl, 2.7 mM KCl, 0.001% (w / v) PX188, pH 7.4 (Cat No. AAV9SP(VB010000-9394npt)-C)) purchased from VectorBuilder was used. To 6.2 μL of each AAV drug substance, 3.1 μL of each of the preparation solutions F, E11, C1, and EC11 shown in Table 3 was added to prepare four samples for each AAV drug substance: a sample with no additional electrolytes or HP-β-CD added (Free), a sample with 100 mM Arg-HCl added (100 mM Arg-HCl), a sample with 100 mM HP-β-CD added (100 mM HP-β-CD), and a sample with 100 mM Arg-HCl and 100 mM HP-β-CD added (100 mM Arg-HCl + 100 mM HP-β-CD).

[0186] The temperature dependence of scattered light intensity when Arg-HCl and HP-β-CD were added alone and simultaneously is shown in Figure 9-1 (AAV2), Figure 9-2 (AAV8), and Figure 9-3 (AAV9). ａｇｇ The results are shown in Table 11-1 (AAV2), Table 11-2 (AAV8), and Table 11-3 (AAV9). When any serotype was used, the T was significantly increased when Arg-HCl or HP-β-CD was added simultaneously compared to when either was added alone. ａｇｇ Change in (ΔT ａｇｇ ) was confirmed to be the largest.

[0187]

[0188]

[0189]

[0190] [Example 10] T in insulin by simultaneous addition of Arg-HCl and HP-β-CD ａｇｇ Solutions containing insulin were prepared under each solvent condition, and the T ａｇｇThe insulin used was human recombinant (yeast) from Roche Diagnostics.

[0191] The temperature dependence of scattered light intensity was compared under four conditions: 1.5 mg / mL insulin, 25 mM sodium phosphate, pH 8.0 (basal condition (free)), and 50 mM Arg-HCl, 50 mM HP-β-CD, or 50 mM Arg-HCl + 50 mM HP-β-CD (free). The T ａｇｇ The results are shown in Table 12. T when Arg-HCl was added alone ａｇｇ Change in (ΔT ａｇｇ ) was 20.1°C, whereas when HP-β-CD was added alone, T ａｇｇ rather decreased, and T ａｇｇ Change in (ΔT ａｇｇ ) was −12.4° C. On the other hand, when Arg-HCl and HP-β-CD were added simultaneously, no clear protein aggregation was detected even at 95° C., which is the upper measurement temperature limit of the instrument.

[0192]

[0193] [Example 11] T in IgG by simultaneous addition of various electrolytes and HP-β-CD ａｇｇ Solutions containing IgG were prepared under each solvent condition, and the T ａｇｇ The IgG used was human plasma-derived IgG from Golden West Diagnostics.

[0194] The basic conditions (free) were 1.5 mg / mL IgG, 5 mM Cit / NaOH, pH 6.0, and various electrolytes (NaCl, KCl, MgCl ２ , CaCl ２ 11-1, 11-2, 11-3, 11-4, 11-5, 11-6, 11-7, 11-8, and 11-9 show the temperature dependence of scattered light intensity when HP-β-CD is added alone and when they are added simultaneously.ａｇｇ The results are shown in Tables 13-1 and 13-2. When comparing at 100 mM, regardless of which electrolyte was used, the T was significantly higher when the electrolyte and HP-β-CD were added simultaneously than when they were added separately. ａｇｇ Change in (ΔT ａｇｇ NaCl, Na-Ace, and Arg-HCl were also compared at 200 mM, but when these electrolytes and HP-β-CD were added simultaneously at 200 mM, no clear protein aggregation was detected even at 95°C, the upper measurement temperature limit of the instrument.

[0195]

[0196]

[0197] [Example 12] T by simultaneous addition of various electrolytes containing Arg in IgG and HP-β-CD ａｇｇ Solutions containing IgG were prepared under each solvent condition, and the T ａｇｇ The IgG used was human plasma-derived IgG from Golden West Diagnostics.

[0198] The basic conditions (free) were 1.5 mg / mL IgG, 5 mM Cit / NaOH, pH 6.0, and the temperature dependence of scattered light intensity was compared when various electrolytes containing Arg (Arg-HCl, Arg-Ace, Arg-Suc, Arg-Asp) and HP-β-CD were added alone or simultaneously to the free solution. ａｇｇ The results are shown in Table 14. When any electrolyte was used, the T was significantly higher when the electrolyte and HP-β-CD were added simultaneously than when they were added separately. ａｇｇ Change in (ΔT ａｇｇ ) was confirmed to be the largest.

[0199]

[0200] [Example 13] T in IgG by simultaneous addition of Arg-HCl and various CDsａｇｇ Solutions containing IgG were prepared under each solvent condition, and the T ａｇｇ The IgG used was human plasma-derived IgG from Golden West Diagnostics.

[0201] The basic conditions (free) were 1.5 mg / mL IgG, 5 mM Cit / NaOH, pH 6.0, and the temperature dependence of scattered light intensity was compared when Arg-HCl and various CDs (HP-α-CD, HP-β-CD, HP-γ-CD) were added alone or simultaneously to the free conditions. The results are shown in Figures 13-1, 13-2, 13-3, and 13-4. ａｇｇ The results are shown in Tables 15-1 and 15-2. When any CD was used, the T was significantly higher when Arg-HCl and CD were added simultaneously than when they were added alone. ａｇｇ Change in (ΔT ａｇｇ When HP-α-CD and HP-β-CD were added simultaneously with Arg-HCl at 200 mM, no clear protein aggregation was detected even at 95°C, the upper measurement temperature limit of the instrument.

[0202]

[0203]

[0204] [Example 14] Effect of the Mixing Ratio of Arg-HCl and HP-β-CD on IgG Solutions containing IgG were prepared under various solvent conditions, and the T values ​​under each solvent condition were measured by the method described in Example 1. ａｇｇ The IgG used was human plasma-derived IgG from Golden West Diagnostics.

[0205] The temperature dependence of scattered light intensity was compared under eight conditions: 1.5 mg / mL IgG, 5 mM Cit / NaOH, pH 6.0 (basic condition), the Arg-HCl concentration was fixed at 100 mM, and the HP-β-CD concentration was varied from 0 to 200 mM. Figure 14-1 shows the T ａｇｇThe results are shown in Table 16-1. As the concentration of HP-β-CD increased, T ａｇｇ Next, the temperature dependence of scattered light intensity was compared under eight conditions: 1.5 mg / mL IgG, 5 mM Cit / NaOH, pH 6.0 as the basic condition (free), the HP-β-CD concentration was fixed at 100 mM, and the Arg-HCl concentration was varied from 0 to 200 mM. Figure 14-2 shows the T ａｇｇ The results are shown in Table 16-2. As the concentration of Arg-HCl increases, T ａｇｇ It was found that the temperature dependence of scattered light intensity increases. In order to investigate the optimal mixture ratio of Arg-HCl and HP-β-CD, the basic conditions (free) were 1.5 mg / mL IgG, 5 mM Cit / NaOH, pH 6.0, and the molar concentration ratio of Arg-HCl and HP-β-CD was changed so that the total molar concentration of Arg-HCl and HP-β-CD was 200 mM. Figure 14-3 shows a comparison of the temperature dependence of scattered light intensity under six conditions. ａｇｇ The results are shown in Table 16-3. From this data, it can be seen that when the molar concentration ratio of Arg-HCl to HP-β-CD is 1:1, T ａｇｇ It was found that the temperature increase was the greatest. Therefore, the molar concentration ratio of Arg-HCl to HP-β-CD was set to 1:1, and the total molar concentration was changed from 120 mM (60 mM Arg-HCl + 60 mM HP-β-CD) to 400 mM (200 mM Arg-HCl + 200 mM HP-β-CD). The temperature dependence of scattered light intensity was compared under eight conditions, and is shown in Figure 14-4. ａｇｇ The results are shown in Table 16-4. From this data, it was found that T ａｇｇ In particular, when the total molar concentration of Arg-HCl and HP-β-CD was 320 mM or higher (160 mM Arg-HCl + 160 mM HP-β-CD), protein aggregation was so suppressed that no clear protein aggregation was detectable even at 95°C, the upper measurement temperature limit of the instrument.

[0206]

[0207]

[0208]

[0209]

[0210] Example 15: Inhibition of soluble protein aggregate formation in IgG by simultaneous addition of Arg-HCl and HP-β-CD. Solutions containing IgG were prepared under various solvent conditions. Human plasma-derived IgG from Golden West Diagnostics was used. The solvent was 5 mM Cit / NaOH, pH 6.0 (basal condition), and 100 mM Arg-HCl, 100 mM HP-β-CD, 50 mM Arg-HCl + 50 mM HP-β-CD, 200 mM Arg-HCl, 200 mM HP-β-CD, 100 mM Arg-HCl + 100 mM HP-β-CD, or 200 mM Arg-HCl + 200 mM HP-β-CD were added to the free solution for a total of eight conditions. The 10 mg / mL IgG solution prepared under each solvent condition was filtered through a 0.22 μm hydrophilic PVDF membrane in a clean bench and then filled into a sterile tube. The percentage (%) of soluble protein aggregates in these IgG solutions was evaluated by size exclusion chromatography (SEC) immediately after preparation (initial) and after 3 months of storage at 25°C. For SEC measurement, a TSKgel G3000SW column (Tosoh Corporation) was used. ＸＬ The HPLC system used was a 7.8 mM ID x 30 cm, 5 μm column (7.8 mM ID x 30 cm, 5 μm) column, consisting of a JASCO liquid delivery unit (PU-2080Plus), a degassing unit (DG-2080-54), a column oven (CO-2060Plus), and a photodiode array detector (MD-2010Plus). The mobile phase consisted of 50 mM sodium phosphate, 300 mM NaCl, 0.5 mg / mL NaN ３ , pH 7.0 was used. A 10 mg / mL IgG solution was diluted with the mobile phase to 1 mg / mL IgG, and 60 μL was injected. Detection was performed by UV absorption at 280 nm. The column temperature was 25°C, and the flow rate was 0.5 mL / min. The percentage (%) of the peak area of ​​the dimer or higher fraction relative to the total peak area was evaluated as the percentage (%) of soluble protein aggregates (Aggregates (%)).

[0211] The results of comparing aggregates (%) under each solvent condition are shown in Figure 15 and Table 17. When 100 mM Arg-HCl or 100 mM HP-β-CD was added alone, the increase in aggregates (%) (ΔAggregates (%)) was 3.1% and 4.0%, respectively, whereas when 50 mM Arg-HCl and 50 mM HP-β-CD were added simultaneously so that the total additive concentration was 100 mM, ΔAggregates (%) was 2.1%, which was lower than the ΔAggregates (%) when Arg-HCl or HP-β-CD was added alone. Furthermore, when 200 mM Arg-HCl or 200 mM HP-β-CD was added alone, the ΔAggregates (%) were 1.7% and 3.4%, respectively. However, when 100 mM Arg-HCl and 100 mM HP-β-CD were added simultaneously so that the total additive concentration was 200 mM, the ΔAggregates (%) was 0.6%, which was lower than the ΔAggregates (%) when Arg-HCl or HP-β-CD was added alone. From the above, the simultaneous addition of Arg-HCl and HP-β-CD was confirmed to have an inhibitory effect on the formation of soluble protein aggregates.

[0212]

[0213] Example 16: Suppression of insoluble protein aggregate formation in IgG by simultaneous addition of Arg-HCl and HP-β-CD. Solutions containing IgG were prepared under various solvent conditions. Human plasma-derived IgG from Golden West Diagnostics was used. The solvent was 5 mM Cit / NaOH, pH 6.0 (basal condition), and 100 mM Arg-HCl, 100 mM HP-β-CD, 50 mM Arg-HCl + 50 mM HP-β-CD, 200 mM Arg-HCl, 200 mM HP-β-CD, 100 mM Arg-HCl + 100 mM HP-β-CD, or 200 mM Arg-HCl + 200 mM HP-β-CD were added to the free solution for a total of eight conditions. The 10 mg / mL IgG solution prepared under each solvent condition was filtered through a 0.22 μm hydrophilic PVDF membrane and then filled into a sterile tube in a clean bench. The concentration of insoluble particles (Particles / mL) in these IgG solutions was evaluated by flow imaging (FI) immediately after preparation (initial) and after 3 months of storage at 25°C. FI measurements were performed using a FlowCam 8100 manufactured by Yokogawa Fluid Imaging Technologies. The flow path of the device was washed with 1% (w / v) Tergazyme (manufactured by Alconox) and PBS(-) at a flow rate of 5 mL / min, and then 1 mL of a 10 mg / mL IgG solution diluted with PBS(-) to 1 mg / mL IgG was placed in the device, and measurements were performed four times under the conditions shown in Table 18. For each particle concentration (Particles / mL) of sizes >2 μm, >5 μm, >10 μm, and >25 μm, the average value and standard deviation of the second, third, and fourth measurement data were calculated. The particle size was calculated as the area-based diameter (ABD).

[0214] The results of comparing the concentration of insoluble microparticles (Particles / mL) under each solvent condition are shown in Figure 16-1 (>2 μm), Figure 16-2 (>5 μm), Figure 16-3 (>10 μm), Figure 16-4 (>25 μm) and Table 19-1 (>2 μm), Table 19-2 (>5 μm), Table 19-3 (>10 μm), and Table 19-4 (>25 μm). Immediately after preparation, all solvent conditions showed approximately the same microparticle concentration for each particle size. On the other hand, after 3 months of storage at 25°C, an increase in the microparticle concentration was observed in the additive-free sample and samples containing Arg-HCl or HP-β-CD alone, whereas almost no increase in microparticles was observed in the sample containing both Arg-HCl and HP-β-CD. The increase in microparticles during storage is thought to be due to insoluble protein aggregates. For example, as shown in Table 19-2, when 100 mM Arg-HCl or 100 mM HP-β-CD was added alone, the concentrations of particles with a size of >5 μm were 357 ± 15 (Particles / mL) and 2230 ± 91 (Particles / mL), respectively. However, when 50 mM Arg-HCl and 50 mM HP-β-CD were added simultaneously so that the total additive concentration was 100 mM, the concentration was 66 ± 7 (Particles / mL), which was lower than the particle concentration (Particles / mL) when Arg-HCl or HP-β-CD was added alone. Furthermore, when 200 mM Arg-HCl or 200 mM HP-β-CD was added alone, the concentration of particles >5 μm in size was 236 ± 56 (Particles / mL) and 1246 ± 22 (Particles / mL), respectively. However, when 100 mM Arg-HCl and 100 mM HP-β-CD were added simultaneously to make the total additive concentration 200 mM, the concentration was 59 ± 7 (Particles / mL), which was lower than the particle concentration (Particles / mL) when Arg-HCl or HP-β-CD was added alone. From the above, the effect of suppressing the formation of insoluble protein aggregates by the simultaneous addition of Arg-HCl and HP-β-CD was confirmed.

[0215]

[0216]

[0217]

[0218]

[0219]

[0220] [Example 22] High concentration of AAV capsids. １２ 5 mL of the AAV capsid dispersion adjusted to capsid particle / mL is added to an Amicon Ultra-15 (50 kDa MWCO; product of MERCK) and ultrafiltered under centrifugation conditions of 1500 × g (however, if concentration requires time, the centrifugal force may be increased to 4000 × g for a swing rotor or 5000 × g for an angle rotor). The above-mentioned preparation solution is added to the AAV capsid dispersion remaining in the membrane supernatant to achieve the desired concentration (for example, 1.0 × 10 １３ If capsid particle / mL, the total volume is 1 mL, 1.0 x 10 １３ The AAV capsid dispersion is recovered by concentrating the AAV capsid in a solution containing 1000 capsid particles / mL (total volume: 0.5 mL). The above concentration method can be replaced by other methods capable of removing the solvent (e.g., TFF, dialysis, drying).

[0221] Using an AAV Titration ELISA Kit (PROGEN), it is confirmed whether the concentration of the AAV capsid dispersion has reached the target concentration. The method for confirming the concentration can be replaced by a method for measuring the nanoparticle concentration (e.g., dynamic light scattering).

[0222] (Note) While the present disclosure has been illustrated using preferred embodiments thereof, it is understood that the scope of the present disclosure should be construed solely by the scope of the claims. It is understood that the patents, patent applications, and other documents cited in this specification are incorporated by reference into this specification in their entirety as if the contents themselves were specifically set forth herein. This application claims priority to Japanese Patent Application No. 2024-097762, filed with the Japan Patent Office on June 17, 2024, the entire contents of which are incorporated by reference herein.

[0223] The disclosed groundbreaking technology is expected to be applied to the medical field.

Claims

1. A composition for preserving proteins, comprising an electrolyte and / or an ion thereof and a cyclic oligosaccharide or a derivative thereof.

2. A composition comprising a protein, an electrolyte and / or its ion, and a cyclic oligosaccharide or a derivative thereof.

3. The composition of claim 2, wherein the composition is for preserving the protein.

4. The composition according to any one of claims 1 to 3, wherein the electrolyte comprises an organic electrolyte.

5. The composition according to any one of claims 1 to 4, wherein the electrolyte comprises one or more selected from inorganic salts, organic salts, organic acids, and organic bases.

6. The composition according to any one of claims 1 to 5, wherein the electrolyte comprises at least one organic acid and at least one organic base.

7. The composition according to any one of claims 1 to 6, which is a solution or a solid.

8. The composition of any one of claims 1 to 7, wherein the electrolyte is present in an amount of 20 millimoles or more per liter of the total composition.

9. The composition of any one of claims 1 to 8, wherein the electrolyte is present in an amount of 100 millimoles or more per liter of the total composition.

10. The composition of any one of claims 1 to 9, wherein the electrolyte comprises an acid selected from acetic acid, citric acid, tartaric acid, glutamic acid, succinic acid, malic acid, and aspartic acid, or a salt thereof.

11. The composition of any one of claims 1 to 10, wherein the electrolyte comprises an organic acid selected from acetic acid, succinic acid, and aspartic acid, or a salt thereof.

12. The composition of any one of claims 1 to 11, wherein the electrolyte comprises acetic acid or a salt thereof.

13. The composition of any one of claims 1 to 12, wherein the electrolyte comprises sodium acetate.

14. The composition of any one of claims 1 to 9, wherein the electrolyte comprises at least one organic base selected from arginine, histidine, and tris(hydroxymethyl)aminomethane, or a salt thereof.

15. The composition of any one of claims 6 to 9 and 14, wherein the organic base is arginine.

16. The composition of any one of claims 1 to 9, wherein the electrolyte comprises at least one selected from sodium chloride, potassium chloride, magnesium chloride, calcium chloride, sodium carbonate, sodium phosphate, and potassium phosphate.

17. The composition of any one of claims 1 to 9 and 16, wherein the electrolyte comprises sodium chloride.

18. The composition of any one of claims 1 to 17, wherein the cyclic oligosaccharide or derivative thereof is selected from substituted or unsubstituted α-CD, β-CD, and γ-CD.

19. The composition of any one of claims 1 to 18, wherein the cyclic oligosaccharide or derivative thereof is hydroxypropyl-β-cyclodextrin (HP-β-CD) or sulfobutylether-β-cyclodextrin (SBE-β-CD).

20. A composition according to any one of claims 1 to 19, wherein the cyclic oligosaccharide or derivative thereof is present at 20 millimoles or more per liter of the total composition.

21. The composition according to any one of claims 1 to 20, wherein the cyclic oligosaccharide or derivative thereof and the electrolyte are contained in equal molar amounts.

22. The composition of any one of claims 1 to 21, wherein the protein is selected from an antibody, an antigen, a hormone, a cytokine, an enzyme, and a capsid.

23. The particle concentration of the capsid is 1 x 10 １３ The composition of claim 22, wherein the concentration is greater than or equal to capsid particles / mL.

24. The composition of any one of claims 1 to 23, which is a pharmaceutical composition.

25. A composition for preserving adeno-associated virus (AAV) capsids, comprising an organic electrolyte and / or ions thereof.

26. A composition for preserving AAV capsids, comprising a cyclic oligosaccharide or a derivative thereof.

27. A composition comprising an AAV capsid and an organic electrolyte or a derivative thereof and / or an ion thereof.

28. A composition comprising an AAV capsid and a cyclic oligosaccharide or a derivative thereof.

29. The composition of claim 25 or claim 27, wherein the organic electrolyte comprises at least one organic base and at least one organic acid.

30. The composition of any one of claims 25, 27, and 29, wherein the organic electrolyte comprises at least one organic base selected from arginine, histidine, and tris(hydroxymethyl)aminomethane.

31. The composition of any one of claims 25, 27, and 29-30, wherein the organic electrolyte comprises at least one organic acid selected from acetic acid, citric acid, tartaric acid, glutamic acid, succinic acid, malic acid, and aspartic acid.

32. The composition of any one of claims 27 and 29 to 31, wherein the organic electrolyte comprises at least one organic base selected from arginine, histidine, and tris(hydroxymethyl)aminomethane, and at least one organic acid selected from acetic acid, citric acid, tartaric acid, glutamic acid, succinic acid, malic acid, and aspartic acid.

33. The composition of any one of claims 25, 27, and 29-32, wherein the organic electrolyte comprises an organic base which is arginine and an organic acid selected from acetic acid, succinic acid, and aspartic acid.

34. The composition of claim 26 or 28, wherein the cyclic oligosaccharide or derivative thereof is selected from substituted or unsubstituted α-CD, β-CD, and γ-CD.

35. The composition of any one of claims 26, 28, and 34, wherein the cyclic oligosaccharide or derivative thereof is hydroxypropyl-β-cyclodextrin (HP-β-CD) or sulfobutylether-β-cyclodextrin (SBE-β-CD).

36. The composition of any one of claims 26, 28 and 34-35, wherein the cyclic oligosaccharide or derivative thereof is present at a concentration of 20 millimoles or more per liter of the total composition.

37. The composition of any one of claims 25, 27, and 29-33, wherein the organic electrolyte comprises an organic base and is present in an amount of 20 millimoles or more per liter of the total composition.

38. The composition of any one of claims 25, 27, and 29-33, wherein the organic electrolyte comprises an organic acid and is present in an amount of 20 millimoles or more per liter of the total composition.

39. The composition of any one of claims 25 to 38, further comprising one or more divalent inorganic salts or carbohydrates.

40. The composition of claim 39, wherein the divalent inorganic salt is selected from magnesium chloride and calcium chloride.

41. The composition of claim 39 or 40, wherein the carbohydrate is selected from glucose, sucrose, trehalose, and mannitol.

42. The particle concentration of the AAV capsid is 1 x 10 １３ The composition according to any one of claims 25 to 41, wherein the capsid particles / mL or more.

43. The composition of any one of claims 25 to 42, further comprising one or more surfactants.

44. The composition of any one of claims 25 to 43, wherein the AAV capsid is an adeno-associated virus vector.

45. A formulation comprising the composition according to any one of claims 1 to 44 filled in a vial or syringe.

46. ​​A kit comprising the composition of any one of claims 1-44.

47. A method for producing the composition according to any one of claims 1 to 44, comprising: 1) providing the protein; and 2) adding and mixing the electrolyte and / or the cyclic oligosaccharide or derivative thereof to the protein.

48. A method for suppressing the formation of aggregates of a protein using the composition according to any one of claims 1 to 44, the method comprising: 1) providing the protein; and 2) placing the electrolyte or ions thereof and / or cyclic oligosaccharides or derivatives thereof under conditions in which they come into contact with the protein.

49. A method for enriching a protein contained in a composition using the composition according to any one of claims 1 to 44, the method comprising: 1) providing the protein; and 2) placing the electrolyte or ions thereof and / or cyclic oligosaccharides or derivatives thereof under conditions in which they come into contact with the protein.

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