WO2021096278A1 - Liquid phase composition for antibody drug - Google Patents

Liquid phase composition for antibody drug Download PDF

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Publication number
WO2021096278A1
WO2021096278A1 PCT/KR2020/015969 KR2020015969W WO2021096278A1 WO 2021096278 A1 WO2021096278 A1 WO 2021096278A1 KR 2020015969 W KR2020015969 W KR 2020015969W WO 2021096278 A1 WO2021096278 A1 WO 2021096278A1
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Prior art keywords
histidine
pharmaceutical composition
composition according
less
antibody
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PCT/KR2020/015969
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French (fr)
Korean (ko)
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강정훈
정소윤
남명주
오인영
김인애
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삼성바이오에피스 주식회사
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Publication of WO2021096278A1 publication Critical patent/WO2021096278A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • A61K39/39591—Stabilisation, fragmentation
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
    • A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
    • A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
    • A61K47/183—Amino acids, e.g. glycine, EDTA or aspartame
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/22—Heterocyclic compounds, e.g. ascorbic acid, tocopherol or pyrrolidones
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/26—Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/08—Solutions
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/32—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against translation products of oncogenes
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin

Definitions

  • the present invention relates to a liquid composition for antibody pharmaceuticals showing excellent stability.
  • Antibodies which are active ingredients of antibody drugs, are obtained by removing impurities such as DNA, cells, and other proteins through the purification process.
  • impurities such as DNA, cells, and other proteins
  • highly purified antibodies are obtained by removing components that can maintain the folded state in a thermodynamically stable form. It may be sensitive to processes such as enzymatic chemical degradation and aggregation.
  • antibody destabilization There are two types of antibody destabilization: chemical types, including changes in covalent bonds, and physical types, including modifications of the three-dimensional spatial structure. Chemical types can include, for example, hydrolysis, oxidation, deamidation, disulfide modification and racemization of antibodies, and physical types include aggregation, adsorption of the antibody, and unfolding that causes a decrease in the activity of the antibody. May be included.
  • the antibody may be modified by physical factors such as surface adsorption and shear, or the activity may be reduced or structural modifications may occur due to modification, oxidation or aggregation of the antibody.
  • the present invention is an anti-Her2 antibody; And it provides a pharmaceutical composition comprising two or more kinds of amino acids, and does not contain acetate.
  • the present invention also provides a pharmaceutical composition comprising an anti-Her2 antibody and two or more amino acids, does not contain acetate, and maintains or improves stability under high temperature storage conditions, freezing/thawing conditions, or stirring conditions.
  • FIG. 1 shows the result of confirming the change in the content of HMW (High Molecular Weight Species) at 40° C. ⁇ 2° C. through size exclusion chromatography (SEC) analysis for the candidate formulation of Example 1.
  • SEC size exclusion chromatography
  • FIG. 2 shows the results of confirming the change in the content of the pertuzumab antibody monomer at 40° C. ⁇ 2° C. through size exclusion chromatography (SEC) analysis for the candidate formulation of Example 1.
  • SEC size exclusion chromatography
  • FIG. 3 shows the results of confirming the change in the content of LMW (Low Molecular Weight Species) at 40° C. ⁇ 2° C. through size exclusion chromatography (SEC) analysis for the candidate formulation of Example 1.
  • SEC size exclusion chromatography
  • FIG. 4 shows the results of confirming the change in the content of HMW at 40° C. ⁇ 2° C. through size exclusion chromatography (SEC) analysis for the candidate formulation of Example 2.
  • SEC size exclusion chromatography
  • Example 5 shows the results of confirming the change in the content of the pertuzumab antibody monomer at 40° C. ⁇ 2° C. through size exclusion chromatography (SEC) analysis for the candidate formulation of Example 2.
  • Example 7 shows the results of confirming the change in the content of HMW under the conditions of 5 cycles of freezing/thawing through size exclusion chromatography (SEC) analysis of the candidate formulation of Example 2.
  • FIG. 15 shows the results of confirming the change in the content of the pertuzumab antibody monomer at 40° C. ⁇ 2° C. through size exclusion chromatography (SEC) analysis for the candidate formulation of Example 4.
  • SEC size exclusion chromatography
  • the present invention anti-Her2 antibody; And to a pharmaceutical composition comprising two or more amino acids and not including acetate.
  • the "formulation" or “pharmaceutical composition” means a mixture or combination of an anti-Her2 antibody, which is an active ingredient capable of exerting a biological effect, and one or more inactive ingredients suitable for therapeutic administration.
  • the formulation may contain one or more pharmaceutically acceptable additional components such that the anti-Her2 antibody stably exhibits activity.
  • the formulation contains two or more amino acids, sugars or sugar alcohols.
  • the "pharmaceutical” (in some cases, “pharmaceutical” or “pharmaceutical”) formulation according to the present invention means a formulation suitable for administration of an antibody so that the desired pharmacological activity can be exhibited without side effects within the scope of medical judgment. I can.
  • the amino acid may be used as a buffer to maintain the pH in a formulation containing an anti-Her2 antibody, or may function as a stabilizer since it may affect the stability of the antibody.
  • the amino acid may be bonded to an antibody, for example, hydrogen bonded, polar bonded, or non-polar bonded.
  • a polar amino acid, a non-polar amino acid, a hydrophilic amino acid and/or a hydrophobic amino acid may be combined to include two or more amino acids.
  • the amino acid may include two or more, for example, two, three, four, five, or more.
  • the pharmaceutical formulation may contain two kinds of amino acids or three kinds of amino acids.
  • the amino acids are, for example, two or more selected from the group consisting of histidine, methionine, cysteine, arginine, glycine, proline, lysine, glutamic acid, isoleucine, phenylalanine, tyrosine, and aspartic acid, for example two , 3, 4, 5 or more amino acids may be included, but are not limited thereto.
  • the amino acid may further include one or more amino acids other than i) histidine and ii) histidine.
  • the amino acid is histidine-methionine, histidine-glycine, histidine-cysteine, histidine-arginine, histidine-proline, histidine-lysine, histidine-glutamic acid, histidine-isoleucine, histidine-phenylalanine, histidine-tyrosine, Histidine-aspartic acid, histidine-methionine-glycine, histidine-methionine-arginine, histidine-methionine-proline, histidine-methionine-lysine, histidine-methionine-glutamic acid, histidine-methionine-isoleucine, histidine-methionine-phenylalanine , Histidine-methionine-tyrosine, or histidine-methionine-aspartic acid.
  • the histidine may be included in a buffer for maintaining the pH in the formulation, or may be included as a stabilizer for maintaining stability by minimizing changes to the anti-Her2 antibody.
  • histidine may be included in the form of a weak acid and a conjugate base or a mixture of a weak base and a conjugate acid.
  • histidine and histidine derivatives such as histidine salts, may be included, for example, histidine chloride, histidine phosphate, or histidine sulfate. Can include.
  • the formulation according to the present invention does not contain histidine acetate.
  • the formulation according to the present invention may include, for example, histidine-HCl, specifically histidine and histidine monohydrochloride monohydrate.
  • the formulation of the present invention may have a pH of 5.0 to 6.5, specifically, 5.0 to 6.0, and more specifically 5.5 to 6.0.
  • the pH may be about 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.5.
  • Amino acids other than histidine may be polar, non-polar, hydrophilic and/or hydrophobic amino acids.
  • Amino acids other than histidine may be selected from the group consisting of, for example, methionine, cysteine, arginine, glycine, proline, lysine, glutamic acid, isoleucine, phenylalanine, tyrosine, and aspartic acid.
  • Amino acids other than histidine may be one or more selected from the group containing methionine, arginine, and glycine. According to a specific embodiment of the present invention, it was confirmed that the formulation containing methionine has high stability under high temperature stress conditions, freezing/thawing conditions, or stirring conditions. Moreover, it was confirmed that methionine can function as an antioxidant.
  • EDTA ethylenediaminetetraacetic acid
  • amino acids other than histidine or amino acids other than histidine and EDTA may be included at the same time.
  • the formulation according to the invention does not contain acetate.
  • “Not including” includes “substantially not including”.
  • “Not including” or “substantially not including” means that acetate is considered a component that does not favor or does not affect the stabilization of the anti-Her2 antibody in the formulation according to the present invention, and is excluded from the component of the formulation. it means.
  • the sugar or sugar alcohol includes the form of a polyol containing a plurality of hydroxyl groups.
  • the sugar or sugar alcohol may function as a stabilizer or tonicity agent in the formulation.
  • the sugar or sugar alcohol is, for example, glucose, sucrose, trehalose, lactose, fructose, maltose, dextran, glycerin, erythritol, arabitol, xylitol, sorbitol, mannitol, melibiose, meletithose, raffinose.
  • Mannotriose, stachiose, maltose, lactulose, maltulose, glucitol, maltitol, lactitol, iso-maltulose may be selected from the group consisting of.
  • the anti-Her2 antibody exhibited stability under high temperature storage conditions, freeze-thaw conditions, or agitation conditions of the antibody in the formulation containing sucrose, sorbitol or trehalose.
  • the formulation may contain, for example:
  • Histidine-HCl glycine, methionine
  • Histidine-HCl glycine, lysine
  • Histidine-HCl glycine, glutamic acid
  • Histidine-HCl glycine, isoleucine
  • Histidine-HCl glycine, phenylalanine
  • Histidine-HCl glycine, tyrosine
  • Histidine-HCl glycine, aspartic acid.
  • the formulation according to the present invention includes (1) histidine-HCl including histidine and histidine monohydrochloride monohydrate, sucrose, methionine, (2) histidine-HCl, sorbitol including histidine and histidine monohydrochloride monohydrate. , Methionine, (3) histidine-HCl including histidine and histidine monohydrochloride monohydrate, trehalose, methionine, (4) histidine-HCl including histidine and histidine monohydrochloride monohydrate, arginine, methionine or (5) ) Histidine and histidine monohydrochloride monohydrate, including histidine-HCl, glycine, and methionine.
  • the molar ratio of sugar or sugar alcohol: anti-Her2 antibody in the formulation according to the present invention may be, for example, 500:1 to 700:1.
  • the molar ratio calculates the molar amount of the anti-Her2 antibody by considering the molecular weight of the anti-Her2 antibody as about 150 kDa, and the known molecular weight of the sugar or sugar alcohol (e.g., sucrose 342.3 g/mol, sorbitol 182.2 g/mol) In consideration of, after calculating the molar amount, the ratio of the molar amount of sugar or sugar alcohol to the molar amount of the anti-Her2 antibody can be calculated and confirmed.
  • the molar ratio of the sugar or sugar alcohol: anti-Her2 antibody may be 500:1 to 700:1, specifically 510:1 to 690:1, or 540:1 to 690:1.
  • the molar ratio of the sugar or sugar alcohol: anti-Her2 antibody is about 500:1, about 501:1, about 502:1, about 503:1, about 504:1, about 505:1, about 506:1, about 507 :1, about 508:1, about 509:1, about 510:1, about 511:1, about 512:1, about 513:1, about 514:1, about 515:1, about 516:1, about 517 :1, about 518:1, about 519:1, about 520:1, about 521:1, about 522:1, about 523:1, about 524:1, about 525:1, about 526:1, about 527 :1, about 528:1, about 529:1, about 530:1, about 531:1, about 532:1, about 533:1, about 534:1, about 535:1, about 536:1, about
  • the formulation according to the present invention may comprise histidine and a salt thereof, for example, histidine: histidine monohydrochloride monohydrate in a molar ratio of 0.2 to 3.0:1.
  • Histidine: Histidine monohydrochloride monohydrate about 0.2, about 0.3, about 0.4, about 0.5, about 0.6, about 0.7, about 0.8, about 0.9, about 1.0, about 1.1, about 1.2, about 1.3, about 1.4, about 1.5 , About 1.6, about 1.7, about 1.8, about 1.9, about 2.0, about 2.1, about 2.2, about 2.3, about 2.4, about 2.5, about 2.6, about 2.7, about 2.8, about 2.9, about 3.0 Value may also be included).
  • the molar ratio calculates the molar amount of histidine taking into account the known molecular weight of histidine, e.g. L-histidine (e.g., 155.2 g/mol histidine), and the known molecular weight of histidine monohydrochloride monohydrate (e.g., 209.6 g/mol) mol), the molar amount is calculated, and then the ratio of the molar amount of histidine monohydrochloride monohydrate to the molar amount of histidine can be calculated and confirmed.
  • L-histidine e.g., 155.2 g/mol histidine
  • histidine monohydrochloride monohydrate e.g., 209.6 g/mol
  • the formulation according to the present invention may contain histidine at a concentration of 5 mM to 50 mM, for example 10 mM to 40 mM, 15 mM to 35 mM, or 20 mM to 30 mM.
  • the histidine is about 5 mM, about 6 mM, about 7 mM, about 8 mM, about 9 mM, about 10 mM, about 11 mM, about 12 mM, about 13 mM, about 14 mM, about 15 mM, about 16 mM , About 17 mM, about 18 mM, about 19 mM, about 20 mM, about 21 mM, about 22 mM, about 23 mM, about 24 mM, about 25 mM, about 26 mM, about 27 mM, about 28 mM, about 29 mM, about 30 mM, about 31 mM, about 32 mM, about 33 mM, about 34 mM, about 35 mM,
  • the formulation includes, for example, 5 mM to 30 mM of amino acids other than histidine selected from the group consisting of methionine, cysteine, arginine, glycine, proline, lysine, glutamic acid, isoleucine, phenylalanine, tyrosine, and aspartic acid. It can be included in concentration. Amino acids other than histidine may be included in a concentration of 5 mM to 30 mM, for example, 7 mM to 20 mM, or 9 mM to 15 mM.
  • the formulation contains 50 mM amino acids other than histidine selected from the group consisting of, for example, methionine, cysteine, arginine, glycine, proline, lysine, glutamic acid, isoleucine, phenylalanine, tyrosine, and aspartic acid.
  • amino acids other than histidine are 50 mM to 150 mM, 60 mM to 140 mM, 70 mM to 135 mM, 80 mM to 135 mM, 90 mM to 135 mM, 100 mM to 140 mM, or 100 mM to 135 mM. It can be included in the concentration of.
  • the amino acid other than histidine is about 50 mM, about 51 mM, about 52 mM, about 53 mM, about 54 mM, about 55 mM, about 55 mM, about 56 mM, about 57 mM, about 58 mM, about 59 mM, about 60 mM, about 61 mM, about 62 mM, about 63 mM, about 64 mM, about 65 mM, about 66 mM, about 67 mM, about 68 mM, about 69 mM, about 70 mM, about 71 mM , About 72 mM, about 73 mM, about 74 mM, about 75 mM, about 76 mM, about 77 mM, about 78 mM, about 79 mM, about 80 mM, about 81 mM, about 82 mM, about 83 mM, about 84 mM, about 85 mM, about
  • the molar ratio of the anti-Her2 antibody: amino acids other than histidine may be 1:10 to 1:100, for example, 1:20 to 1:80, or 1:30 to 1:70.
  • the molar ratio of the anti-Her2 antibody: amino acids other than histidine is about 1:10, about 1:11, about 1:12, about 1:13, about 1:14, about 1:15, about 1:16 , About 1:17, about 1:18, about 1:19, about 1:20, about 1:21, about 1:22, about 1:23, about 1:24, about 1:25, about 1:26 , About 1:27, about 1:28, about 1:29, about 1:30, about 1:31, about 1:32, about 1:33, about 1:34, about 1:35, about 1:36 , About 1:37, about 1:38, about 1:39, about 1:40, about 1:41, about 1:42, about 1:43, about 1:44, about 1:45, about 1:46 , About 1:47, about 1:48, about 1:49, about 1:50, about 1:51
  • the molar ratio of amino acids other than histidine: anti-Her2 antibody may be 300:1 to 450:1, specifically 330:1 to 420:1, or 350:1 to 400:1.
  • the molar ratio of amino acids other than histidine: anti-Her2 antibody is about 300:1, about 301:1, about 302:1, about 303:1, about 304:1, about 305:1, about 306:1, about 307 :1, about 308:1, about 309:1, about 310:1, about 311:1, about 312:1, about 313:1, about 314:1, about 315:1, about 316:1, about 317 :1, about 318:1, about 319:1, about 320:1, about 321:1, about 322:1, about 323:1, about 324:1, about 325:1, about 326:1, about 327 :1, about 328:1, about 329:1, about 330:1, about 331:1, about 332:1, about 333:1, about 334:1, about 335:1, about 335
  • the molar ratio of amino acids other than histidine: anti-Her2 antibody may be 500:1 to 700:1, specifically 510:1 to 690:1, or 540:1 to 690:1.
  • the molar ratio of amino acids other than histidine: anti-Her2 antibody is about 500:1, about 501:1, about 502:1, about 503:1, about 504:1, about 505:1, about 506:1, about 507 :1, about 508:1, about 509:1, about 510:1, about 511:1, about 512:1, about 513:1, about 514:1, about 515:1, about 516:1, about 517 :1, about 518:1, about 519:1, about 520:1, about 521:1, about 522:1, about 523:1, about 524:1, about 525:1, about 526:1, about 527 :1, about 528:1, about 529:1, about 530:1, about 531:1, about 532:1, about 533:1, about 534:1, about 535:1, about
  • the calculation for the molar ratio is the same as described above.
  • the molar amount of the anti-Her2 antibody is calculated by considering the molecular weight of the anti-Her2 antibody as about 150 kDa, and the molar amount is calculated by considering the known molecular weight of amino acids other than histidine, and then amino acids other than histidine are calculated based on the molar amount of the anti-Her2 antibody. It can be checked by calculating the ratio of the molar amount.
  • the formulation may contain 50 mM to 150 mM sugar or sugar alcohol.
  • the sugar or sugar alcohol is 50 mM to 150 mM, 60 mM to 140 mM, 70 mM to 135 mM, 80 mM to 135 mM, 90 mM to 135 mM, 100 mM to 140 mM, or 100 mM to 135 mM It can be included in the concentration of.
  • the sugar or sugar alcohol is about 50 mM, about 51 mM, about 52 mM, about 53 mM, about 54 mM, about 55 mM, about 55 mM, about 56 mM, about 57 mM, about 58 mM, about 59 mM, about 60 mM, about 61 mM, about 62 mM, about 63 mM, about 64 mM, about 65 mM, about 66 mM, about 67 mM, about 68 mM, about 69 mM, about 70 mM, about 71 mM , About 72 mM, about 73 mM, about 74 mM, about 75 mM, about 76 mM, about 77 mM, about 78 mM, about 79 mM, about 80 mM, about 81 mM, about 82 mM, about 83 mM, about 84 mM, about 85 mM, about 80 m
  • concentration ranges of 5 to 50 mM, 50 mM to 150 mM, and 5 mM to 30 mM it can be understood that the formulation of the present invention includes all of any concentrations within each range.
  • the formulation may further include a bulking agent, a viscosity modifier, a stabilizer, a surfactant, an antioxidant, or a tonicity agent.
  • the formulation according to the present invention may include, for example, polysorbate, poloxamer, sodium dodecyl sulfate (SDS), sodium laurel sulfate (SLS), and the like.
  • the polysorbate is polysorbate 20, polysorbate 21, polysorbate 40, polysorbate 60, polysorbate 61, polysorbate 65, polysorbate 80, polysorbate 81, polysorbate 85 or its It may contain a mixture.
  • the formulation according to the present invention may comprise a polysorbate, for example polysorbate 20.
  • the formulation according to the present invention may comprise a polysorbate in a concentration of 0.01 to 0.1% (w/v) or 0.02 to 0.08% (w/v).
  • Formulations according to the invention are about 0.01% (w/v), about 0.02% (w/v), about 0.03% (w/v), about 0.04% (w/v), about 0.05% (w/v) , About 0.06% (w/v), about 0.07% (w/v), or about 0.08% (w/v) (values between listed values may also be included).
  • the anti-Her2 antibody may be, for example, pertuzumab.
  • Pertuzumab herein is a monoclonal antibody that targets HER2 and is marketed as Perjeta®.
  • Pertuzumab binds to region II of the extracellular matrix domain of the HER2 receptor and inhibits the formation of dimers, thereby inhibiting the receptor-mediated signaling pathway.
  • ErbB2 also known as HER2/neu, is the second member of the EGFR family and exhibits biological functions by forming a heterodimer with the other three members of the EGFR family, EGFR/HER1, HER3 and HER4.
  • the neu gene encoding ErbB2 was first isolated from rat neuroblastoma.
  • the homologous gene of the neu gene in human somatic cells is located on chromosome 17q21.1.
  • the encoded product is ErbB2, consisting of 1255 amino acids having a molecular weight of approximately 185 kDa, and the 720 to 987 amino acid positions belong to the tyrosine kinase active domain.
  • ErbB2 is dependent on the PI3K and MAPK signaling pathways, can reduce the expression of cyclin D and c-myc, and decreases the expression of the cyclin-dependent kinase (cdk) inhibitor p27kipl, inducing cell proliferation by inhibiting cdk2 activity. do.
  • Antibody means a protein capable of neutralizing, blocking, inhibiting, eliminating, reducing or interfering with a biological activity.
  • monoclonal antibody is meant an antibody molecule of single molecular composition obtained from a population of substantially the same antibody, ie the individual antibodies occupying the population are identical except for possible naturally occurring mutations that may be present in trace amounts.
  • Monoclonal antibodies are highly specific, so they are directed against a single antigenic site and exhibit a single binding specificity and affinity for a particular epitope.
  • the antibody includes not only the complete antibody form, but also the antigen-binding fragment of the antibody.
  • a complete antibody is a tetrameric protein of about 150 kDa with two full-length light chains and two full-length heavy chains, each of which is linked to a heavy chain by a disulfide bond.
  • Antigen-binding fragments or antibody fragments of an antibody may include Fab, F(ab'), F(ab')2, Fv, or an epitope-binding fragment thereof.
  • Fab is a structure having a variable region of a light and heavy chain, a constant region of a light chain, and a first constant region (CH1) of a heavy chain, and has one antigen-binding site.
  • F(ab') differs from Fab in that it has a hinge region containing one or more cysteine residues at the C-terminus of the heavy chain CH1 domain.
  • F(ab')2 antibodies are produced by disulfide bonds between cysteine residues in the hinge region of F(ab').
  • Fv fragments are antibody fragments that contain a complete antibody recognition and binding site.
  • the double-chain Fv (two-chain Fv) is a non-covalent bond where the heavy chain variable region and the light chain variable region are connected, and the single-chain Fv (scFv) is generally a heavy chain variable region and a light chain variable region through a peptide linker. It is connected by this covalent bond.
  • Heavy chain may refer to the heavy chain of an IgG antibody, and refers to both a full-length heavy chain including a variable region domain VH and three constant region domains CH1, CH2 and CH3 for conferring specificity to an antigen, and fragments thereof.
  • light chain as used herein may mean the light chain of an IgG antibody, and all of the full-length light chain including the variable region domain VL and the constant region domain CL for conferring specificity to the antigen and fragments thereof it means.
  • Variable domain refers to the light and heavy chain portions of an antibody molecule comprising the amino acid sequences of the complementarity determining regions (CDRs; ie, CDR1, CDR2, and CDR3), and framework regions (FR).
  • CDRs complementarity determining regions
  • FR framework regions
  • VH refers to the variable domain of the heavy chain.
  • VL refers to the variable domain of the light chain.
  • CDR complementarity determining region
  • the “skeletal region” (FR) is a variable domain residue other than a CDR residue.
  • Each variable domain typically has 4 FRs identified as FR1, FR2, FR3 and FR4.
  • the heavy chain constant region has gamma ( ⁇ ), mu ( ⁇ ), alpha ( ⁇ ), delta ( ⁇ ) and epsilon ( ⁇ ) types, and has subclasses of gamma 1 ( ⁇ 1), gamma 2 ( ⁇ 2), and gamma 3 ( ⁇ 3). ), gamma4( ⁇ 4), alpha1( ⁇ 1) and alpha2( ⁇ 2).
  • the constant region of the light chain has kappa ( ⁇ ) and lambda ( ⁇ ) types.
  • Epitope refers to a determinant to which an antibody can specifically bind. Epitopes are usually composed of a group of chemically active surface molecules, such as amino acids or sugar side chains, and generally have specific three-dimensional structural characteristics as well as specific charge characteristics.
  • Monoclonal antibodies refer to antibodies obtained from a population of substantially homogeneous antibodies, i.e., those that are identical except for possible naturally occurring mutations in which the individual antibodies occupying the population may be present in trace amounts. Monoclonal antibodies are highly specific and are thus directed against a single antigenic site.
  • a non-human (eg murine) antibody in its “humanized” form is a chimeric antibody that contains a minimal sequence derived from a non-human immunoglobulin.
  • Humanized antibodies are those of a non-human species (donor antibody), such as a mouse, rat, rabbit or non-human primate, that retain the desired specificity, affinity and ability of the hypervariable region residues of the recipient. It is a replaced human immunoglobulin (receptor antibody).
  • the heavy chain CDR, light chain CDR, VH and VL sequences of Pertuzumab according to the present invention are described in US7862817B2 or US7537931B2, which documents can be incorporated by reference.
  • unpaired cysteine variants of pertuzumab or VHS extension variants of pertuzumab including acidic variants or disulfide-reducing variants of pertuzumab may be included, and US8652474B2 or US9181346B2, acidic
  • the unpaired cysteine variants of Pertuzumab, including variants or disulfide reducing variants are described in US9815904B2, US9969811B, and VHS extension variants of Pertuzumab in US7560111B2, US8241630B2 or US7879325B2, respectively, This document may also be incorporated by reference.
  • Preparation of an anti-Her2 antibody involves a conventional antibody preparation method, e.g., inserting a nucleic acid encoding a pertuzumab antibody into a replicable vector, transfecting the vector into cells, and then culturing the cells and culturing them. It can be prepared through a method of recovering the antibody from the cell.
  • a method for preparing a Pertuzumab antibody is specifically described in US 7879325 or US 8241630, and the document can be incorporated by reference.
  • Perturb comprising a sequence that is 85% or more, specifically 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more, or 100% identical to the sequence Zumab antibodies may be included.
  • the identity of the sequences can be determined by comparing two sequences that are optimally aligned, and additions or deletions (i.e., gaps) compared to a reference sequence (not including additions or deletions) for optimal alignment of the two sequences. It may include.
  • Identity is expressed as a percentage, and the percentage determines the number of positions in which the same nucleic acid base or amino acid residue appears in both sequences to generate the number of matched positions, and the number of matched positions is divided by the total number of positions, and the It can be calculated by multiplying the result by 100.
  • the percent sequence identity can be determined, for example, using the Needle program, the BLOSUM62 matrix program, and the like.
  • the anti-Her2 antibody may be included in an amount effective for effective treatment or prevention of a disease. Accordingly, the anti-Her2 antibody may be included in a concentration of 20 mg/ml to 150 mg/ml. In the formulation according to the present invention, the anti-Her2 antibody may be 25 mg/ml to 130 mg/ml, 30 mg/ml to 110 mg/ml, or 40 mg/ml to 100 mg/ml.
  • the anti-Her2 antibody is about 20 mg/ml, about 21 mg/ml, about 22 mg/ml, about 23 mg/ml, about 24 mg/ml, about 25 mg/ml, about 26 mg/ml, About 27 mg/ml, about 28 mg/ml, about 29 mg/ml, about 30 mg/ml, about 31 mg/ml, about 32 mg/ml, about 33 mg/ml, about 34 mg/ml, about 35 mg/ml, about 36 mg/ml, about 37 mg/ml, about 38 mg/ml, about 39 mg/ml, about 40 mg/ml, about 41 mg/ml, about 42 mg/ml, about 43 mg/ ml, about 44 mg/ml, about 45 mg/ml, about 46 mg/ml, about 47 mg/ml, about 48 mg/ml, about 49 mg/ml, about 50 mg/ml, about 51 mg/ml, About 52 mg/ml, about 53 mg/ml, about 54 mg/ml, about
  • the present invention may be a formulation for IV administration containing an anti-Her2 antibody at a concentration of 20 mg/ml to 30 mg/ml.
  • an anti-Her2 antibody at a concentration of 20 mg/ml to 30 mg/ml.
  • the present invention relates to a formulation containing an anti-Her2 antibody and two or more amino acids, no acetate, and maintaining or improving stability under high temperature storage conditions, freezing/thawing conditions, or stirring conditions.
  • the formulation according to the present invention may further contain sugar or sugar alcohol.
  • the sugar or sugar alcohol is, for example, glucose, sucrose, trehalose, lactose, fructose, maltose, dextran, glycerin, erythritol, arabitol, xylitol, sorbitol, mannitol, melibiose, meletithose, raffinose.
  • Mannotriose, stachiose, maltose, lactulose, maltulose, glucitol, maltitol, lactitol, iso-maltulose may be selected from the group consisting of.
  • the anti-Her2 antibody exhibited stability under high temperature storage conditions, freeze-thaw conditions, or agitation conditions of the antibody in the formulation containing sucrose, sorbitol or trehalose.
  • a “stable” formulation is one that maintains the physical stability, chemical stability or biological activity of the anti-Her2 antibody from mechanical stress, thermal stress, or stress resulting from freezing and thawing, at a selected temperature for a selected period of time, for example about 5 ⁇ At least about 1, 2, 3, 4, 5, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks at 3°C, about 25 ⁇ 2°C or about 40 ⁇ 2°C , 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 Stable for a period of weeks or longer, or after freezing (eg -20°C or -70°C) and thawing, eg, after freezing and thawing in 1, 2, 3, 4, 5, or more cycles Or, it means that it is stable under agitation stress conditions, for example, 100 rpm or more, 200 rpm or more, 300 rpm
  • Assessment of aggregation formation e.g., using size exclusion chromatography or measuring turbidity and/or visual inspection), cation exchange chromatography, evaluation of charge heterogeneity using image capillary isoelectric focusing (icIEF) or capillary zone electrophoresis, amino- Terminal or carboxy-terminal sequence analysis, mass spectrometric analysis, SDS-PAGE analysis comparing reduced and intact antibodies, peptide map (e.g., trypsin or LYS-C) analysis, or biological activity or antigen of the antibody Stability can be evaluated through various methods including evaluation of binding function and the like.
  • image capillary isoelectric focusing (icIEF) or capillary zone electrophoresis evaluation of charge heterogeneity using image capillary isoelectric focusing (icIEF) or capillary zone electrophoresis
  • amino- Terminal or carboxy-terminal sequence analysis e.g., mass spectrometric analysis
  • SDS-PAGE analysis comparing reduced and intact antibodies
  • UV light scattering or size exclusion chromatography SEC
  • the antibody is not modified by cleavage or bond formation, it may mean that the antibody remains chemically stable. Chemical stability can be ascertained by detecting and quantifying the chemically modified form of the antibody.
  • the biological activity of the antibody by antigen binding assay or neutralization assay is about 70% to about 150%, about 80% to about 125%, or about 90% to about 110% of the antibody biological activity at the time of preparation of the formulation
  • the biological activity is It can mean keeping.
  • the biological activity of the antibody under certain conditions and times is 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 100% of the antibody biological activity at the time of preparation of the formulation, If it is 105%, 110%, 115%, 120% or 125%, biological activity can be maintained.
  • the biological activity of an anti-Her2 antibody can be ascertained, for example, through the level of Her2 binding. Specifically, the biological activity of the anti-Her2 antibody can be measured through Receptor-Binding Activity (RBA).
  • RBA Receptor-Binding Activity
  • the Her2 binding level measured by confirming the receptor binding activity under storage conditions of 40°C ⁇ 2°C for 8 weeks in the formulation according to the present invention may be, for example, 92% or more, specifically 92 to 97%.
  • the Her2 binding level measured by confirming the receptor-binding activity under storage conditions of 40°C ⁇ 2°C for 12 weeks may be, for example, 90% or more, specifically 90 to 95%.
  • the Her2 binding level measured by confirming the receptor binding activity under storage conditions of 5°C ⁇ 3°C for 8 weeks in the formulation according to the present invention may be, for example, 97% or more, specifically 97 to 99%.
  • the Her2 binding level measured by confirming the receptor binding activity in the formulation of -70 ⁇ 10° C. 1 cycle or more, for example 5 cycles of freeze/thaw cycle in the formulation according to the present invention is, for example, 97 % Or more, specifically 97 to 99%.
  • the stability of the antibody in the formulation can be determined by identifying aggregation or fragment levels in the formulation. It can be determined by measuring whether a high molecular weight product (HMW) and/or a low molecular weight species (LMW) are formed in the formulation. The content of the high molecular weight product and/or the low molecular weight product in the formulation can be measured under certain temperature conditions. The content of the high molecular weight product and/or the low molecular weight product can be measured by size exclusion HPLC (SE-HPLC).
  • HMW high molecular weight product
  • LMW low molecular weight species
  • the stability of the antibody in the formulation can be determined by measuring whether a high molecular weight product (HMW) is formed in the formulation.
  • HMW high molecular weight product
  • the amount of change in HMW in the formulation can be measured under specific temperature conditions, and the HMW can be measured by size exclusion HPLC (SE-HPLC).
  • the% content of HMW measured by size exclusion HPLC was 6.0% or less, 5.0% or less, 4.0% or less, 3.0% or less, 2.0% or less, 1.0% or less, 0.9% or less, 0.8% It may be 0.7% or less, 0.6% or less, 0.5% or less, 0.4% or less, 0.3% or less, 0.2% or less, or 0.1% or less.
  • the HMW measured by size exclusion HPLC (SE-HPLC) at 40° C. ⁇ 2° C. for 1 week or more in the formulation according to the present invention may be 0.5% or less.
  • the% of HMW content measured through size exclusion HPLC (SE-HPLC) in the formulation under storage conditions at a temperature of 40° C. ⁇ 2° C. for 1 week is, for example, 0.27% or less, specifically 0.16 to 0.27%.
  • the content% of HMW measured through size exclusion HPLC (SE-HPLC) in the formulation under storage conditions at a temperature of 40°C ⁇ 2°C for 2 weeks may be, for example, 0.33% or less, specifically 0.17 to 0.33%.
  • the content% of HMW measured through size exclusion HPLC (SE-HPLC) in the formulation under storage conditions at a temperature of 40°C ⁇ 2°C for 4 weeks may be, for example, 0.4% or less, specifically 0.20 to 0.4%.
  • the content% of HMW measured through size exclusion HPLC (SE-HPLC) in the formulation under storage conditions at a temperature of 40°C ⁇ 2°C for 8 weeks may be, for example, 0.48% or less, and specifically 0.24 to 0.48%.
  • the HMW content% measured through size exclusion HPLC (SE-HPLC) in the formulation at a temperature of 40° C. ⁇ 2° C. for 1 week storage conditions is, for example, 0.2% or less, specifically 0.15 to 0.2%.
  • the content% of HMW measured through size exclusion HPLC (SE-HPLC) in the formulation at a temperature of 40°C ⁇ 2°C for 2 weeks storage may be, for example, 0.25% or less, specifically 0.2 to 0.25%.
  • the content% of HMW measured through size exclusion HPLC (SE-HPLC) in the formulation under storage conditions at a temperature of 40°C ⁇ 2°C for 8 weeks may be, for example, 0.5% or less, specifically 0.35 to 0.5%.
  • the content% of HMW measured through size exclusion HPLC (SE-HPLC) in the formulation under storage conditions at a temperature of 40°C ⁇ 2°C for 12 weeks may be, for example, 0.6% or less, and specifically 0.35 to 0.6%.
  • HMW measured by size exclusion HPLC (SE-HPLC) in the formulation of 1 cycle or more freeze/thaw cycle is 2.0% or less, 1.0% or less, 0.9% or less, 0.8% or less, 0.7% Or less, 0.6% or less, 0.5% or less, 0.4% or less, 0.3% or less, 0.25% or less, 0.2% or less, 0.15% or less, or 0.1% or less.
  • HMW measured through size exclusion HPLC (SE-HPLC) under 400 rpm stirring conditions may be 0.25% or less, 0.2% or less, 0.15% or less, or 0.1% or less.
  • the% content of HMW measured via size exclusion HPLC (SE-HPLC) in the formulation of 1 cycle or more, eg 5 cycles of freeze/thaw cycles is, for example, 2.0 % Or less, 1.0% or less, 0.9% or less, 0.8% or less, 0.7% or less, 0.6% or less, 0.5% or less, 0.4% or less, 0.3% or less, 0.25% or less, 0.2% or less, specifically 0.1 to 0.2% I can.
  • the content% of HMW measured by size exclusion HPLC (SE-HPLC) in the formulation at 400 rpm stirring conditions is, for example, 2.0% or less, 1.0% or less, 0.9% or less, 0.8% or less, 0.7% It may be less than, 0.6% or less, 0.5% or less, 0.4% or less, 0.3% or less, 0.25% or less, specifically 0.15 to 0.25%.
  • the content% of the antibody monomer in the formulation may be 96% or more, 97% or more, 98% or more, 99% or more, or 100%.
  • the content of the antibody monomer may be 96% or more, 97% or more, 98% or more, 99% or more, or 100%.
  • the percentage of the IgG content in the formulation measured by CE-SDS (Capillary electrophoresis with sodium dodecylsulfate) at 40°C ⁇ 2°C for 8 weeks is, for example, 90% or more, specifically 90% to 93%.
  • the amount of IgG in the formulation measured through CE-SDS at 40° C. ⁇ 2° C. for 12 weeks may be, for example, 87% or more, and specifically 87% to 89%.
  • the% of the IgG content in the formulation measured through CE-SDS at a temperature of 5°C ⁇ 3°C for 8 weeks may be, for example, 95% or more, specifically 95% to 97%.
  • the percentage of the IgG content in the formulation measured through CE-SDS under conditions of 5°C ⁇ 3°C for 12 weeks may be, for example, 94% or more, and specifically 94% to 95%.
  • the percentage of IgG content measured through CE-SDS in the formulation of 1 cycle or more, eg, 5 cycles of freeze/thaw cycle is, for example, 95% or more, specifically It may be 95% to 97%.
  • the stability of the antibody in the formulation can be determined by measuring whether a variant in the antibody is formed.
  • the variant may mean, for example, an antibody having an amino acid sequence different from that of the main antibody, and may include substitution, deletion, and/or addition of amino acids at a specific position within or adjacent to the amino acid sequence of the antibody.
  • the variant may include a change in charge due to modification of the antibody, and may be, for example, a charge variant.
  • Charged variants can be acidic and have low pI values compared to unmodified antibodies. Acidic variants can be produced when they have a negative charge or lose a positive charge than the main antibody.
  • the charge variant may exhibit basicity compared to an unmodified antibody and may have a high pI value. Basic variants can be produced if they have a positive charge or lose a negative charge than the main antibody.
  • These charge variants can occur by oxidation of antibodies, deamidation, processing of the C-terminus of the lysine residue, formation of pyroglutamate at the N-terminus, and non-enzymatic glycosylation.
  • the charge variants may be separated by ion exchange chromatography, for example, cation exchange chromatography to separate proteins according to charge, or charge heterogeneity may be confirmed by image capillary isoelectric focusing (icIEF).
  • the content% of acidic variant or basic variant measured by cation exchange HPLC during the period is 50% or less, 45% or less, 40% or less, 35% or less, 30% or less, 25% or less, 20% or less, 19% or less, It may be 18% or less, 17% or less, 15% or less, or 10% or less.
  • icIEF Imaged Capillary Isoelectric Focusing
  • the content% of the acidic variant may be 70% or less under conditions of 1 week or more.
  • the content% of the acidic variant in the formulation measured by icIEF at a temperature of 40° C. ⁇ 2° C. for 2 weeks storage may be, for example, 45% or less, and specifically 44.0 to 44.5%.
  • the content% of the acidic variant in the formulation measured by icIEF at a temperature of 40°C ⁇ 2°C for 4 weeks storage may be, for example, 53% or less, specifically 50 to 53%.
  • the content% of the acidic variant in the formulation measured by icIEF at a temperature of 40°C ⁇ 2°C for 8 weeks storage may be, for example, 65% or less, specifically 60 to 65%.
  • the content% of the acidic variant in the formulation measured by icIEF at a temperature of 40°C ⁇ 2°C for 12 weeks storage may be, for example, 70% or less, specifically 65 to 70%.
  • the content% of acidic variant or basic variant in the formulation of 1 cycle or more of freeze/thaw cycle is 20% or less, 18% or less, 17% or less, 15% or less, 10% or less, 5% or less, 4% Or less, 3% or less, 2% or less, and 1% or less.
  • the content% of acidic variant or basic variant in the formulation under 400 rpm stirring conditions is 20% or less, 18% or less, 17% or less, 15% or less, 10% or less, 5% or less, 4% or less, 3% or less, 2% It is less than or equal to 1%.
  • the content% of the acidic variant in the formulation under the conditions of -70 °C ⁇ 10 °C freeze/thaw 5 cycles is, for example, 38% or less, specifically 36 to It could be 38%.
  • the formulation according to the present invention can be evaluated as stable for at least 1 month, at least 2 months, at least 3 months, or at least 6 months at 5°C ⁇ 3°C or more, particularly 40°C ⁇ 2°C.
  • the formulation according to the present invention can be evaluated as stable after freezing and thawing at -70°C ⁇ 10°C, freezing and thawing at -70 ⁇ 10°C for 24 hours, and then thawing at room temperature for 90 minutes Including, the cycle may be repeated 1-5 times, and maintained for 72 hours at a temperature of -70 ⁇ 10°C in the fifth cycle.
  • it can be evaluated as stable after stirring under the stirring conditions of 400 rpm.
  • the anti-Her2 antibody may be included in a therapeutically effective amount.
  • Effective amount refers to an amount necessary or sufficient to bind to region II of the extracellular matrix domain of the HER2 receptor to inhibit the formation of dimers, thereby inhibiting receptor-mediated signaling pathways. For example, it is the amount necessary to prevent or treat various morphological symptoms of conditions associated with dimer formation of HER2.
  • the symptom is cancer, it may be an amount necessary for reducing the number of cancer cells, reducing the size of the tumor, inhibiting invasion of cancer cells, inhibiting tumor metastasis, inhibiting tumor growth, or reducing symptoms of cancer.
  • An effective amount can prolong the survival period without disease progression, increase the overall survival period, or ameliorate adverse symptoms caused by cancer.
  • the effective amount may vary depending on the subject's height, weight, type or severity of the disease.
  • An effective amount can be determined and administered to a subject. As well as divided doses, it may be administered daily or continuously in alternating doses. The dose may be increased or decreased proportionally depending on the purpose of treatment, drugs used in combination, and the like.
  • Treatment includes the reduction or relief of one or more symptoms associated with or caused by the condition, disorder or disease being treated.
  • treatment can reduce one or more symptoms of the disorder or can completely eliminate the disorder.
  • the level of administration of the anti-Her2 antibody can vary in an amount effective for the desired therapeutic response through the patient, without toxicity to the patient. Dosage is the activity of the anti-Her2 antibody in the formulation, route of administration, time of administration, duration of treatment, other drugs and/or compounds in combination, age, sex, weight, symptoms, general health condition, and previous medical history of the patient to be treated. It can be determined through various factors including.
  • Anti-Her2 antibodies can be administered in combination with other Her2 target antibodies or chemotherapeutic agents. Concomitant administration includes co-administration and sequential administration using separate formulations or single pharmaceutical formulations. Other Her2 target antibodies or chemotherapeutic agents may be administered before or after administration of the anti-Her2 antibody. Other Her2 target antibodies or chemotherapeutic agents can be administered concurrently with administration of the anti-Her2 antibody.
  • Her2 target antibodies may be antibodies that bind to HER2 and inhibit the growth of cancer cells that overexpress HER2, and may be, for example, trastzumab (Herceptin®).
  • Chemotherapeutic agents are compounds useful in the treatment of cancer, such as anthracyclines, alkylating agents (e.g., mitomycin C), alkyl sulfonates, aziridines, ethylenimine, methylmelamine, nitrogen mustard, nitrosourea, antibiotics. , Antimetabolites, folic acid analogs (e.g., dihydrofolate reductase inhibitors such as methotrexate), purine analogs, pyrimidine analogs, enzymes, podophyllotoxin, or platinum-containing agents, and the like.
  • alkylating agents e.g., mitomycin C
  • alkyl sulfonates e.g., aziridines, ethylenimine, methylmelamine, nitrogen mustard, nitrosourea, antibiotics.
  • Antimetabolites folic acid analogs (e.g., dihydrofolate reductase inhibitors such as methotrexate), purine analogs
  • Antibody dosages can generally be from about 0.1 mg/kg to about 50 mg/kg. When administering a chemotherapeutic agent, it is usually administered at a known dosage, or a lower amount of the antibody may be administered due to side effects resulting from the administration of the chemotherapeutic agent or the combined action of drugs.
  • the dosing schedule and preparation for the chemotherapeutic agent can be used according to the manufacturer's instructions, or can be determined experimentally by a skilled person.
  • compositions are administered to a patient by, for example, intravenous administration, intramuscular, intraperitoneal, cerebrospinal, subcutaneous, intraarticular, intrasynovial, intrathecal, oral, topical, or inhalation routes. It is preferred to administer intravenously, intramuscularly or subcutaneously, and the pharmaceutical formulation of the present invention may be administered intravenously.
  • Syringes for subcutaneous administration Injection devices (eg INJECT-EASETM and GENJECTTM devices); Syringe pens (eg GENPENTM); Needleless devices (eg MEDIJECTORTM and BIOJECTORTM);
  • a subcutaneous patch delivery system can be used.
  • Articles of manufacture may include bottles, vials (eg double chamber vials), syringes (eg double chamber syringes) or tubular containers.
  • the container can be made of glass or plastic.
  • the container contains the formulation of the present invention, and instructions for use may be indicated on the container or on a label affixed to the container.
  • the article of manufacture may further include package inserts indicating instructions for further configuration and use, such as other buffers, diluents, fillers, needles, and syringes.
  • Pertuzumab antibody formulation was prepared as in Table 1. With regard to the sequence of the Pertuzumab antibody, reference may be made to the specific antibody sequence described in US7862817B2 or US7537931B2. Regarding the acidic variant of Pertuzumab, US8652474B2 or US9181346B2, the unpaired cysteine variants of Pertuzumab including the acidic variant or the reducing disulfide variant are US9815904B2, US9969811B, VHS extension variants of Pertuzumab (VHS extension variants). For more information, reference may be made to US7560111B2, US8241630B2 or US7879325B2.
  • Pertuzumab antibody was concentrated to 36 mg/mL or more, and pH 4.0-5.0 acetate (acetate), pH 5.0-6.0 histidine-HCl, pH 6.0-7.0 phosphate (phosphate) was used in the concentrated DS (drug substance). , Divided into 3 times at 3, 5, and 10 hour intervals and dialyzed in a 2-8°C cold chamber, pH 4.0-5.0 acetate (acetate), pH 5.0-6.0 histidine-HCl, pH 6.0- Exchange was carried out with each of 7.0 phosphates (phosphates).
  • the pH range of the sample was selected according to the intrinsic characteristics of the three components, and Samples 3 and 4 were the same as pH 5.0, and Samples 6 and 7 were the same as pH 6.0, and the average value was described by repeating three experiments.
  • the commercially available control drug Perjeta® contains histidine-acetate, and the average value was recorded by repeating three experiments.
  • Example 1-1 The sample of Table 1 prepared in Example 1-1 was filled into a vial by 0.5 mL, and samples were placed in a chamber for 1 week, 2 weeks, and 4 weeks in a 40°C ⁇ 2°C stability chamber. -in), and chamber-out the sample at each time point.
  • SE-HPLC Size Exclusion High Performance Liquid Chromatography: Tosoh Bioscience
  • HMW High molecular weight species
  • monomer and LMW high molecular weight species
  • monomers monomers
  • LMW Low molecular weight species
  • Example 1-2 it was confirmed that the composition containing histidine-HCl at pH 5.0-6.0 has better stability than the composition containing acetate or phosphate, and exhibits equivalent or higher stability compared to Perjeta®. .
  • a pertuzumab formulation according to Table 3 was prepared in the same manner as in Example 1-1.
  • Example 1-2 in order to further study the formulations containing histidine-HCl of Samples 4 to 6, which were effective in terms of %HMW, the same pH as Perjeta® was fixed to 6.0, and as an additional component, a predetermined concentration of sugar, The stability of the composition comprising a sugar alcohol or a second amino acid was compared.
  • the other composition was the same, but histidine-HCl was replaced with succinate (succinate), and the results were compared.
  • Samples 1, 6, and Perjeta® were of the same composition except that succinate, histidine-HCl, and histidine-acetate were included, and the average value was confirmed by performing 3 repeated experiments.
  • 1 week, 2 By confirming the experimental results up to weeks, 4 weeks and 8 weeks, it was attempted to confirm a little more long-term stability.
  • Example 2-1 The sample of Table 3 prepared in Example 2-1 was filled into a vial with 0.5 mL each, and samples for 1 week, 2 weeks, 4 weeks, and 8 weeks in a 40°C ⁇ 2°C stability chamber were each timed. Chamber-out at the point. Using SE-HPLC, HMW, monomer and LMW ratios were analyzed for each sample under the conditions of 40° C. ⁇ 2° C. initial, 1 week, 2 weeks, 4 weeks and 8 weeks. The analysis results are shown in Table 4 and FIGS. 4 to 6.
  • samples 6 to 10 containing sucrose, trehalose, sorbitol, arginine, and glycine along with histidine-HCl had less %HMW compared to Perjeta®, and thus were more stable.
  • samples 6 to 10 containing sucrose, trehalose, sorbitol, arginine, and glycine along with histidine-HCl had less %HMW compared to Perjeta®, and thus were more stable.
  • samples 6 to 10 containing sucrose, trehalose, sorbitol, arginine, and glycine along with histidine-HCl had less %HMW compared to Perjeta®, and thus were more stable.
  • Example 7 A formulation according to Table 7 was prepared in the same manner as in Example 1-1. Among the additional ingredients having good effects in Example 2, the concentration of sucrose, sorbitol, and glycine was changed, and methionine, a third amino acid, was added thereto to confirm the formulation stability. The samples were subjected to 3 repeated experiments to confirm the result as an average value, and compared with the Perjeta® experimental value of Example 2.
  • Table 8 shows the concentrations of L-histidine and L-histidine monohydrochloride monohydrate contained in the sample containing 20 mM histidine-HCl to adjust the pH to 5.5, 6.0, and 6.5, respectively.
  • Example 3-1 The sample of Table 7 prepared in Example 3-1 was filled into a vial with 0.5 mL each, and samples for 1 week, 2 weeks, 4 weeks, and 8 weeks were each time in a 40°C ⁇ 2°C stability chamber. Chamber-out at the point. Using SE-HPLC, HMW, monomer and LMW ratios were analyzed for each sample under the conditions of 40° C. ⁇ 2° C. initial, 1 week, 2 weeks, 4 weeks and 8 weeks. The analysis results are shown in Table 9 and FIGS. 9 to 11.
  • HMW, monomer and LMW ratios were analyzed for each sample of Table 12 under the conditions of 40°C ⁇ 2°C initial, 1 week, 2 weeks, 4 weeks and 8 weeks. The analysis results are shown in Table 13 and FIGS. 14 to 16.
  • Sample 13 (Example 3) were confirmed to be more stable due to less %HMW, and among them, 3 than Sample 10 (Example 2), Sample 3 (Example 3) and Sample 8 (Example 3) containing two kinds of amino acids. It was confirmed that Sample 13 (Example 3) containing species of amino acids had the least %HMW and was the most stable.
  • Sample 13 (Example 3) were confirmed to be more stable due to less %HMW, and among them, 3 than Sample 10 (Example 2), Sample 3 (Example 3) and Sample 8 (Example 3) containing two kinds of amino acids. It was confirmed that Sample 13 (Example 3) containing species of amino acids had the least %HMW and was the most stable.

Abstract

The present invention relates to a pharmaceutical composition including an anti-Her2 antibody and exhibiting excellent stability.

Description

항체 의약품용 액상 조성물Liquid composition for antibody pharmaceuticals
본 발명은 우수한 안정성을 나타내는 항체 의약품용 액상 조성물에 관한 것이다.The present invention relates to a liquid composition for antibody pharmaceuticals showing excellent stability.
항체 의약품의 유효성분인 항체는 정제 과정을 통해 DNA, 세포, 기타 다른 단백질 등의 불순물이 제거되어 얻어지는데, 특히 고도로 정제된 항체는 열역학적으로 안정한 형태로 접힌 상태를 유지할 수 있는 성분이 제거되어, 효소 화학적 분해 및 응집 등의 과정에 민감할 수 있다. Antibodies, which are active ingredients of antibody drugs, are obtained by removing impurities such as DNA, cells, and other proteins through the purification process.In particular, highly purified antibodies are obtained by removing components that can maintain the folded state in a thermodynamically stable form. It may be sensitive to processes such as enzymatic chemical degradation and aggregation.
항체의 불안정성 (destabilization)에는 공유결합에서의 변화를 포함하는 화학적 유형 및 3차원 공간 구조의 변형을 포함하는 물리적 유형이 있다. 화학적 유형에는 예를 들어 항체의 가수분해, 산화, 탈아미드화, 이황화 변형 및 라세미화 (racemization)가 포함될 수 있으며, 물리적 유형에는 항체의 응집, 흡착 및 항체의 활성 감소를 유발하는 풀림 (unfolding)이 포함될 수 있다. There are two types of antibody destabilization: chemical types, including changes in covalent bonds, and physical types, including modifications of the three-dimensional spatial structure. Chemical types can include, for example, hydrolysis, oxidation, deamidation, disulfide modification and racemization of antibodies, and physical types include aggregation, adsorption of the antibody, and unfolding that causes a decrease in the activity of the antibody. May be included.
사용 전 보관 및 운송하는 기간 동안 표면 흡착 및 전단 (shear)과 같은 물리적 요소에 의해 항체가 변형되거나, 항체의 변형, 산화 또는 응집에 의해 활성이 감소되거나 구조적 변형이 발생할 수 있다. During storage and transport prior to use, the antibody may be modified by physical factors such as surface adsorption and shear, or the activity may be reduced or structural modifications may occur due to modification, oxidation or aggregation of the antibody.
따라서, 항체를 가공하고 취급하는 과정에서 발생하는 다양한 요소 및 환경으로부터 항체의 구조를 안정화시킬 필요가 있고, 이를 위해 특히 항체를 안정화할 수 있는 제형에 대한 요구가 있다. Accordingly, there is a need to stabilize the structure of the antibody from various factors and environments generated in the process of processing and handling the antibody, and for this purpose, there is a need for a formulation capable of stabilizing the antibody in particular.
발명의 요약Summary of the invention
본 발명의 목적은 항-Her2 항체를 안정화할 수 있는 약제학적 조성물을 제공하는 것이다.It is an object of the present invention to provide a pharmaceutical composition capable of stabilizing an anti-Her2 antibody.
상기 목적을 달성하기 위하여, 본 발명은 항-Her2 항체; 및 2종 이상의 아미노산을 포함하고, 아세트산염을 포함하지 않는 약제학적 조성물을 제공한다. In order to achieve the above object, the present invention is an anti-Her2 antibody; And it provides a pharmaceutical composition comprising two or more kinds of amino acids, and does not contain acetate.
본 발명은 또한, 항-Her2 항체 및 2종 이상의 아미노산을 포함하고, 아세트산염을 포함하지 않으며, 고온 보관 조건, 냉동/해동 조건 또는 교반 조건에서 안정성이 유지 또는 향상된 약제학적 조성물을 제공한다. The present invention also provides a pharmaceutical composition comprising an anti-Her2 antibody and two or more amino acids, does not contain acetate, and maintains or improves stability under high temperature storage conditions, freezing/thawing conditions, or stirring conditions.
도 1은 실시예 1의 후보 제형에 대하여 크기 배제 크로마토그래피 (SEC) 분석을 통해 40℃±2℃ 온도 조건에서 HMW (High Molecular Weight Species)의 함량 변화를 확인한 결과를 나타낸 것이다. FIG. 1 shows the result of confirming the change in the content of HMW (High Molecular Weight Species) at 40° C.±2° C. through size exclusion chromatography (SEC) analysis for the candidate formulation of Example 1. FIG.
도 2는 실시예 1의 후보 제형에 대하여 크기 배제 크로마토그래피 (SEC) 분석을 통해 40℃±2℃ 온도 조건에서 퍼투주맙 항체 단량체의 함량 변화를 확인한 결과를 나타낸 것이다. FIG. 2 shows the results of confirming the change in the content of the pertuzumab antibody monomer at 40° C.±2° C. through size exclusion chromatography (SEC) analysis for the candidate formulation of Example 1. FIG.
도 3은 실시예 1의 후보 제형에 대하여 크기 배제 크로마토그래피 (SEC) 분석을 통해 40℃±2℃ 온도 조건에서 LMW (Low Molecular Weight Species)의 함량 변화를 확인한 결과를 나타낸 것이다. FIG. 3 shows the results of confirming the change in the content of LMW (Low Molecular Weight Species) at 40° C.±2° C. through size exclusion chromatography (SEC) analysis for the candidate formulation of Example 1. FIG.
도 4는 실시예 2의 후보 제형에 대하여 크기 배제 크로마토그래피 (SEC) 분석을 통해 40℃±2℃ 온도 조건에서 HMW의 함량 변화를 확인한 결과를 나타낸 것이다. FIG. 4 shows the results of confirming the change in the content of HMW at 40° C.±2° C. through size exclusion chromatography (SEC) analysis for the candidate formulation of Example 2. FIG.
도 5는 실시예 2의 후보 제형에 대하여 크기 배제 크로마토그래피 (SEC) 분석을 통해 40℃±2℃ 온도 조건에서 퍼투주맙 항체 단량체의 함량 변화를 확인한 결과를 나타낸 것이다. 5 shows the results of confirming the change in the content of the pertuzumab antibody monomer at 40° C.±2° C. through size exclusion chromatography (SEC) analysis for the candidate formulation of Example 2.
도 6은 실시예 2의 후보 제형에 대하여 크기 배제 크로마토그래피 (SEC) 분석을 통해 40℃±2℃ 온도 조건에서 LMW의 함량 변화를 확인한 결과를 나타낸 것이다. 6 shows the results of confirming the change in the content of LMW in the temperature condition of 40°C±2°C through size exclusion chromatography (SEC) analysis of the candidate formulation of Example 2.
도 7은 실시예 2의 후보 제형에 대하여 크기 배제 크로마토그래피 (SEC) 분석을 통해 냉동/해동 5 사이클 조건에서 HMW의 함량 변화를 확인한 결과를 나타낸 것이다. 7 shows the results of confirming the change in the content of HMW under the conditions of 5 cycles of freezing/thawing through size exclusion chromatography (SEC) analysis of the candidate formulation of Example 2.
도 8은 실시예 2의 후보 제형에 대하여 크기 배제 크로마토그래피 (SEC) 분석을 통해 교반 조건에서 HMW의 함량 변화를 확인한 결과를 나타낸 것이다. 8 shows the results of confirming the change in the content of HMW under stirring conditions through size exclusion chromatography (SEC) analysis of the candidate formulation of Example 2.
도 9는 실시예 3의 후보 제형에 대하여 크기 배제 크로마토그래피 (SEC) 분석을 통해 40℃±2℃ 온도 조건에서 HMW의 함량 변화를 확인한 결과를 나타낸 것이다. 9 shows the results of confirming the change in the content of HMW under the temperature condition of 40°C±2°C through size exclusion chromatography (SEC) analysis of the candidate formulation of Example 3.
도 10은 실시예 3의 후보 제형에 대하여 크기 배제 크로마토그래피 (SEC) 분석을 통해 40℃±2℃ 온도 조건에서 퍼투주맙 항체 단량체의 함량 변화를 확인한 결과를 나타낸 것이다. 10 shows the results of confirming the change in the content of the pertuzumab antibody monomer at 40°C±2°C through size exclusion chromatography (SEC) analysis for the candidate formulation of Example 3.
도 11은 실시예 3의 후보 제형에 대하여 크기 배제 크로마토그래피 (SEC) 분석을 통해 40℃±2℃ 온도 조건에서 LMW의 함량 변화를 확인한 결과를 나타낸 것이다. 11 shows the results of confirming the change in the content of LMW in the temperature condition of 40°C±2°C through size exclusion chromatography (SEC) analysis of the candidate formulation of Example 3.
도 12는 실시예 3의 후보 제형에 대하여 크기 배제 크로마토그래피 (SEC) 분석을 통해 냉동/해동 5 사이클 조건에서 HMW의 함량 변화를 확인한 결과를 나타낸 것이다. 12 shows the results of confirming the change in the content of HMW under the conditions of 5 cycles of freezing/thawing through size exclusion chromatography (SEC) analysis of the candidate formulation of Example 3.
도 13은 실시예 3의 후보 제형에 대하여 크기 배제 크로마토그래피 (SEC) 분석을 통해 교반 조건에서 HMW의 함량 변화를 확인한 결과를 나타낸 것이다. 13 shows the results of confirming the change in the content of HMW under stirring conditions through size exclusion chromatography (SEC) analysis of the candidate formulation of Example 3.
도 14는 실시예 4의 후보 제형에 대하여 크기 배제 크로마토그래피 (SEC) 분석을 통해 40℃±2℃ 온도 조건에서 HMW의 함량 변화를 확인한 결과를 나타낸 것이다. 14 shows the results of confirming the change in the content of HMW in the temperature condition of 40° C.±2° C. through size exclusion chromatography (SEC) analysis of the candidate formulation of Example 4.
도 15는 실시예 4의 후보 제형에 대하여 크기 배제 크로마토그래피 (SEC) 분석을 통해 40℃±2℃ 온도 조건에서 퍼투주맙 항체 단량체의 함량 변화를 확인한 결과를 나타낸 것이다. FIG. 15 shows the results of confirming the change in the content of the pertuzumab antibody monomer at 40° C.±2° C. through size exclusion chromatography (SEC) analysis for the candidate formulation of Example 4. FIG.
도 16은 실시예 4의 후보 제형에 대하여 크기 배제 크로마토그래피 (SEC) 분석을 통해 40℃±2℃ 온도 조건에서 LMW의 함량 변화를 확인한 결과를 나타낸 것이다. 16 shows the results of confirming the change in the content of LMW in the temperature condition of 40°C±2°C through size exclusion chromatography (SEC) analysis for the candidate formulation of Example 4.
도 17은 실시예 4의 후보 제형에 대하여 크기 배제 크로마토그래피 (SEC) 분석을 통해 교반 조건에서 HMW의 함량 변화를 확인한 결과를 나타낸 것이다. 17 shows the results of confirming the change in the content of HMW under stirring conditions through size exclusion chromatography (SEC) analysis of the candidate formulation of Example 4.
발명의 상세한 설명 및 바람직한 구현예Detailed description and preferred embodiments of the invention
다른 식으로 정의되지 않는 한, 본 명세서에서 사용된 모든 기술적 및 과학적 용어들은 본 발명이 속하는 기술분야에서 숙련된 전문가에 의해서 통상적으로 이해되는 것과 동일한 의미를 갖는다. 일반적으로, 본 명세서에서 사용된 명명법은 본 기술분야에서 잘 알려져 있고 통상적으로 사용되는 것이다.Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by an expert skilled in the art to which the present invention belongs. In general, the nomenclature used in this specification is well known and commonly used in the art.
본 발명은 일 관점에서, 항-Her2 항체; 및 2종 이상의 아미노산을 포함하고, 아세트산염을 포함하지 않는 약제학적 조성물에 관한 것이다.In one aspect, the present invention, anti-Her2 antibody; And to a pharmaceutical composition comprising two or more amino acids and not including acetate.
본 발명에 따른 "제형" 또는 "약제학적 조성물"은 생물학적 효과를 발휘할 수 있는 유효성분인 항-Her2 항체와 치료용 투여에 적합한 하나 이상의 비활성성분의 혼합 또는 조합을 의미한다. 상기 제형은 항-Her2 항체가 안정적으로 활성을 나타낼 수 있도록 약제학적으로 허용되는 하나 이상의 추가 구성을 포함할 수 있다. 상기 제형은 2종 이상의 아미노산, 당 또는 당 알코올을 포함한다. The "formulation" or "pharmaceutical composition" according to the present invention means a mixture or combination of an anti-Her2 antibody, which is an active ingredient capable of exerting a biological effect, and one or more inactive ingredients suitable for therapeutic administration. The formulation may contain one or more pharmaceutically acceptable additional components such that the anti-Her2 antibody stably exhibits activity. The formulation contains two or more amino acids, sugars or sugar alcohols.
본 발명에 따른"약학"(경우에 따라서 "약제학적" 또는 "약학적") 제형은 의학적 판단의 범위 내에서 부작용 없이 목적하는 바람직한 약리 활성을 나타낼 수 있도록, 항체의 투여에 적합한 제형을 의미할 수 있다. The "pharmaceutical" (in some cases, "pharmaceutical" or "pharmaceutical") formulation according to the present invention means a formulation suitable for administration of an antibody so that the desired pharmacological activity can be exhibited without side effects within the scope of medical judgment. I can.
상기 아미노산은 항-Her2 항체를 포함하는 제형에서 pH를 유지하는 버퍼로 사용되거나, 항체의 안정성에 영향을 미칠 수 있으므로 안정화제로 기능할 수 있다. 상기 아미노산은 항체와 결합 예를 들어, 수소결합, 극성결합 또는 비극성결합할 수 있다. 극성 아미노산, 비극성 아미노산, 친수성 아미노산 및/또는 소수성 아미노산을 조합하여 2종 이상의 아미노산을 포함할 수 있다. The amino acid may be used as a buffer to maintain the pH in a formulation containing an anti-Her2 antibody, or may function as a stabilizer since it may affect the stability of the antibody. The amino acid may be bonded to an antibody, for example, hydrogen bonded, polar bonded, or non-polar bonded. A polar amino acid, a non-polar amino acid, a hydrophilic amino acid and/or a hydrophobic amino acid may be combined to include two or more amino acids.
하나의 실시예에서, 상기 아미노산은 2종 이상, 예를 들어 2종, 3종, 4종, 5종 또는 그 이상을 포함할 수 있다. 본 발명에 따른 구체적 실시예에서, 상기 약학 제형은 2종의 아미노산, 또는 3종의 아미노산을 포함할 수 있다.In one embodiment, the amino acid may include two or more, for example, two, three, four, five, or more. In a specific embodiment according to the present invention, the pharmaceutical formulation may contain two kinds of amino acids or three kinds of amino acids.
상기 아미노산은 예를 들어, 히스티딘, 메티오닌, 시스테인, 알지닌, 글리신, 프롤린, 라이신, 글루탐산, 이소루이신, 페닐알라닌, 티로신, 아스파트산으로 구성된 군에서 선택되는 2종 이상, 예를 들어 2종, 3종, 4종, 5종 또는 그 이상의 아미노산을 포함할 수 있으나, 이에 제한되는 것은 아니다. The amino acids are, for example, two or more selected from the group consisting of histidine, methionine, cysteine, arginine, glycine, proline, lysine, glutamic acid, isoleucine, phenylalanine, tyrosine, and aspartic acid, for example two , 3, 4, 5 or more amino acids may be included, but are not limited thereto.
하나의 구체예에서, 상기 아미노산은 예를 들어, i) 히스티딘 및 ii) 히스티딘 이외의 아미노산을 추가로 1종 이상 포함할 수 있다. In one embodiment, the amino acid may further include one or more amino acids other than i) histidine and ii) histidine.
예를 들어, 상기 아미노산은 히스티딘-메티오닌, 히스티딘-글리신, 히스티딘-시스테인, 히스티딘-알지닌, 히스티딘-프롤린, 히스티딘-라이신, 히스티딘-글루탐산, 히스티딘-이소루이신, 히스티딘-페닐알라닌, 히스티딘-티로신, 히스티딘-아스파트산, 히스티딘-메티오닌-글리신, 히스티딘-메티오닌-알지닌, 히스티딘-메티오닌 -프롤린, 히스티딘-메티오닌-라이신, 히스티딘-메티오닌-글루탐산, 히스티딘-메티오닌-이소루이신, 히스티딘-메티오닌-페닐알라닌, 히스티딘-메티오닌-티로신, 또는 히스티딘-메티오닌-아스파트산을 포함할 수 있다.For example, the amino acid is histidine-methionine, histidine-glycine, histidine-cysteine, histidine-arginine, histidine-proline, histidine-lysine, histidine-glutamic acid, histidine-isoleucine, histidine-phenylalanine, histidine-tyrosine, Histidine-aspartic acid, histidine-methionine-glycine, histidine-methionine-arginine, histidine-methionine-proline, histidine-methionine-lysine, histidine-methionine-glutamic acid, histidine-methionine-isoleucine, histidine-methionine-phenylalanine , Histidine-methionine-tyrosine, or histidine-methionine-aspartic acid.
상기 히스티딘은 제형 중 pH를 유지하는 버퍼에 포함되거나, 항-Her2 항체에 대한 변화를 최소화하여 안정성을 유지하기 위한 안정화제로 포함될 수 있다. 경우에 따라서, 히스티딘은 약산과 짝염기 또는 약염기와 짝산의 혼합물 형태로 포함될 수 있다. 하나의 구체예에서, 상기 제형 중에 히스티딘 및 히스티딘 유도체 예를 들어, 히스티딘 염의 형태로 포함될 수 있고, 예를 들어 히스티딘 클로라이드 (histidine chloride), 히스티딘 포스페이트 (histidine phosphate), 또는 히스티딘 설페이트 (histidine sulfate)를 포함할 수 있다. 본 발명에 따른 제형은 히스티딘 아세테이트 (histidine acetate)를 포함하지 않는다. The histidine may be included in a buffer for maintaining the pH in the formulation, or may be included as a stabilizer for maintaining stability by minimizing changes to the anti-Her2 antibody. In some cases, histidine may be included in the form of a weak acid and a conjugate base or a mixture of a weak base and a conjugate acid. In one embodiment, in the formulation, histidine and histidine derivatives, such as histidine salts, may be included, for example, histidine chloride, histidine phosphate, or histidine sulfate. Can include. The formulation according to the present invention does not contain histidine acetate.
본 발명에 따른 제형은 예를 들어 히스티딘-HCl, 구체적으로 히스티딘 및 히스티딘 모노하이드로클로라이드 일수화물 (histidine monohydrochloride monohydrate)을 포함할 수 있다. The formulation according to the present invention may include, for example, histidine-HCl, specifically histidine and histidine monohydrochloride monohydrate.
이에 따른 본 발명의 제형은 pH가 5.0 내지 6.5, 구체적으로, 5.0 내지 6.0, 더 구체적으로 5.5 내지 6.0일 수 있다. 구체적으로, pH는 약 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.5일 수 있다. Accordingly, the formulation of the present invention may have a pH of 5.0 to 6.5, specifically, 5.0 to 6.0, and more specifically 5.5 to 6.0. Specifically, the pH may be about 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.5.
상기 히스티딘 이외의 아미노산은 극성, 비극성, 친수성 및/또는 소수성 아미노산일 수 있다. 상기 히스티딘 이외의 아미노산은 예를 들어, 메티오닌, 시스테인, 알지닌, 글리신, 프롤린, 라이신, 글루탐산, 이소루이신, 페닐알라닌, 티로신, 아스파트산으로 구성된 군에서 선택될 수 있다.Amino acids other than histidine may be polar, non-polar, hydrophilic and/or hydrophobic amino acids. Amino acids other than histidine may be selected from the group consisting of, for example, methionine, cysteine, arginine, glycine, proline, lysine, glutamic acid, isoleucine, phenylalanine, tyrosine, and aspartic acid.
상기 히스티딘 이외의 아미노산은 메티오닌, 알지닌 및 글리신을 포함하는 군에서 선택되는 하나 이상일 수 있다. 본 발명의 구체적 실시예에 따르면, 메티오닌을 포함하는 제형은 고온 스트레스 조건, 냉동/해동 조건, 또는 교반조건에서 안정성이 높음을 확인하였다. 더욱이, 메티오닌은 항산화제로서 기능할 수 있음을 확인하였다.Amino acids other than histidine may be one or more selected from the group containing methionine, arginine, and glycine. According to a specific embodiment of the present invention, it was confirmed that the formulation containing methionine has high stability under high temperature stress conditions, freezing/thawing conditions, or stirring conditions. Moreover, it was confirmed that methionine can function as an antioxidant.
경우에 따라서, 히스티딘 이외의 아미노산을 대신하여 EDTA (ethylenediaminetetraacetic acid)를 포함하거나, 히스티딘 이외의 아미노산과 EDTA를 동시에 포함할 수 있다. In some cases, EDTA (ethylenediaminetetraacetic acid) may be included in place of amino acids other than histidine, or amino acids other than histidine and EDTA may be included at the same time.
본 발명에 따른 제형은 아세트산염을 포함하지 않는다. "포함하지 않음"은 "실질적으로 포함하지 않음"을 포함한다. "포함하지 않음" 또는 "실질적으로 포함하지 않음"은 본 발명에 따른 제형에서 아세트산염을 항-Her2 항체의 안정화에 유리하지 않거나, 영향을 미치지 않는 성분으로 고려하고, 제형의 성분에서 배제함을 의미한다. The formulation according to the invention does not contain acetate. "Not including" includes "substantially not including". "Not including" or "substantially not including" means that acetate is considered a component that does not favor or does not affect the stabilization of the anti-Her2 antibody in the formulation according to the present invention, and is excluded from the component of the formulation. it means.
상기 당 또는 당 알코올은 복수의 하이드록실기를 포함하는 폴리올의 형태를 포함한다. 상기 당 또는 당 알코올은 제형 중 안정화제 또는 등장화제(tonicity agents)로 기능할 수 있다.The sugar or sugar alcohol includes the form of a polyol containing a plurality of hydroxyl groups. The sugar or sugar alcohol may function as a stabilizer or tonicity agent in the formulation.
상기 당 또는 당 알코올은 예를 들어, 글루코스, 수크로오스, 트레할로스, 락토오스, 프럭토오스, 말토오스, 덱스트란, 글리세린, 에리트리톨, 아라비톨, 자일리톨, 솔비톨, 만니톨, 멜리바이오스, 멜레치토스, 라피노오스, 만노트리오스, 스타키오스, 말토오스, 락툴로오스, 말툴로오스, 글루시톨, 말티톨, 락티톨, 이소-말툴로오스로 구성된 군에서 선택될 수 있다. The sugar or sugar alcohol is, for example, glucose, sucrose, trehalose, lactose, fructose, maltose, dextran, glycerin, erythritol, arabitol, xylitol, sorbitol, mannitol, melibiose, meletithose, raffinose. , Mannotriose, stachiose, maltose, lactulose, maltulose, glucitol, maltitol, lactitol, iso-maltulose may be selected from the group consisting of.
본 발명에 따른 구체예에서, 수크로오스, 솔비톨 또는 트레할로스를 포함하는 제형에서 항체의 고온 보관 조건, 냉동-해동 조건 또는 교반 조건 중 항-Her2 항체가 안정성을 나타냄을 확인하였다. In the embodiment according to the present invention, it was confirmed that the anti-Her2 antibody exhibited stability under high temperature storage conditions, freeze-thaw conditions, or agitation conditions of the antibody in the formulation containing sucrose, sorbitol or trehalose.
상기 제형은 예를 들어, 다음을 포함할 수 있다:The formulation may contain, for example:
히스티딘-HCl, 수크로오스, 메티오닌; Histidine-HCl, sucrose, methionine;
히스티딘-HCl, 수크로오스, 시스테인;Histidine-HCl, sucrose, cysteine;
히스티딘-HCl, 수크로오스, 알지닌; Histidine-HCl, sucrose, arginine;
히스티딘-HCl, 수크로오스, 프롤린;Histidine-HCl, sucrose, proline;
히스티딘-HCl, 수크로오스, 라이신;Histidine-HCl, sucrose, lysine;
히스티딘-HCl, 수크로오스, 글루탐산; Histidine-HCl, sucrose, glutamic acid;
히스티딘-HCl, 수크로오스, 이소루이신; Histidine-HCl, sucrose, isoleucine;
히스티딘-HCl, 수크로오스, 페닐알라닌; Histidine-HCl, sucrose, phenylalanine;
히스티딘-HCl, 수크로오스, 티로신; Histidine-HCl, sucrose, tyrosine;
히스티딘-HCl, 수크로오스, 아스파트산; Histidine-HCl, sucrose, aspartic acid;
히스티딘-HCl, 솔비톨, 메티오닌;Histidine-HCl, sorbitol, methionine;
히스티딘-HCl, 솔비톨, 시스테인;Histidine-HCl, sorbitol, cysteine;
히스티딘-HCl, 솔비톨, 알지닌; Histidine-HCl, sorbitol, arginine;
히스티딘-HCl, 솔비톨, 프롤린;Histidine-HCl, sorbitol, proline;
히스티딘-HCl, 솔비톨, 라이신;Histidine-HCl, sorbitol, lysine;
히스티딘-HCl, 솔비톨, 글루탐산; Histidine-HCl, sorbitol, glutamic acid;
히스티딘-HCl, 솔비톨, 이소루이신; Histidine-HCl, sorbitol, isoleucine;
히스티딘-HCl, 솔비톨, 페닐알라닌; Histidine-HCl, sorbitol, phenylalanine;
히스티딘-HCl, 솔비톨, 티로신; Histidine-HCl, sorbitol, tyrosine;
히스티딘-HCl, 솔비톨, 아스파트산;Histidine-HCl, sorbitol, aspartic acid;
히스티딘-HCl, 트레할로스, 메티오닌;Histidine-HCl, trehalose, methionine;
히스티딘-HCl, 트레할로스, 시스테인;Histidine-HCl, trehalose, cysteine;
히스티딘-HCl, 트레할로스, 알지닌; Histidine-HCl, trehalose, arginine;
히스티딘-HCl, 트레할로스, 프롤린;Histidine-HCl, trehalose, proline;
히스티딘-HCl, 트레할로스, 라이신;Histidine-HCl, trehalose, lysine;
히스티딘-HCl, 트레할로스, 글루탐산; Histidine-HCl, trehalose, glutamic acid;
히스티딘-HCl, 트레할로스, 이소루이신; Histidine-HCl, trehalose, isoleucine;
히스티딘-HCl, 트레할로스, 페닐알라닌; Histidine-HCl, trehalose, phenylalanine;
히스티딘-HCl, 트레할로스, 티로신; Histidine-HCl, trehalose, tyrosine;
히스티딘-HCl, 알지닌, 메티오닌;Histidine-HCl, arginine, methionine;
히스티딘-HCl, 알지닌, 시스테인;Histidine-HCl, arginine, cysteine;
히스티딘-HCl, 알지닌, 프롤린;Histidine-HCl, arginine, proline;
히스티딘-HCl, 알지닌, 라이신;Histidine-HCl, arginine, lysine;
히스티딘-HCl, 알지닌, 글루탐산; Histidine-HCl, arginine, glutamic acid;
히스티딘-HCl, 알지닌, 이소루이신; Histidine-HCl, arginine, isoleucine;
히스티딘-HCl, 알지닌, 페닐알라닌; Histidine-HCl, arginine, phenylalanine;
히스티딘-HCl, 알지닌, 티로신; Histidine-HCl, arginine, tyrosine;
히스티딘-HCl, 트레할로스, 아스파트산;Histidine-HCl, trehalose, aspartic acid;
히스티딘-HCl, 글리신, 메티오닌; Histidine-HCl, glycine, methionine;
히스티딘-HCl, 글리신, 시스테인;Histidine-HCl, glycine, cysteine;
히스티딘-HCl, 글리신, 알지닌; Histidine-HCl, glycine, arginine;
히스티딘-HCl, 글리신, 프롤린;Histidine-HCl, glycine, proline;
히스티딘-HCl, 글리신, 라이신;Histidine-HCl, glycine, lysine;
히스티딘-HCl, 글리신, 글루탐산; Histidine-HCl, glycine, glutamic acid;
히스티딘-HCl, 글리신, 이소루이신; Histidine-HCl, glycine, isoleucine;
히스티딘-HCl, 글리신, 페닐알라닌; Histidine-HCl, glycine, phenylalanine;
히스티딘-HCl, 글리신, 티로신; 또는Histidine-HCl, glycine, tyrosine; or
히스티딘-HCl, 글리신, 아스파트산.Histidine-HCl, glycine, aspartic acid.
구체적으로, 본 발명에 따른 제형은 (1) 히스티딘 및 히스티딘 모노하이드로클로라이드 일수화물을 포함하는 히스티딘-HCl, 수크로오스, 메티오닌, (2) 히스티딘 및 히스티딘 모노하이드로클로라이드 일수화물을 포함하는 히스티딘-HCl, 솔비톨, 메티오닌, (3) 히스티딘 및 히스티딘 모노하이드로클로라이드 일수화물을 포함하는 히스티딘-HCl, 트레할로스, 메티오닌, (4) 히스티딘 및 히스티딘 모노하이드로클로라이드 일수화물을 포함하는 히스티딘-HCl, 알지닌, 메티오닌 또는 (5) 히스티딘 및 히스티딘 모노하이드로클로라이드 일수화물을 포함하는 히스티딘-HCl, 글리신, 메티오닌을 포함할 수 있다.Specifically, the formulation according to the present invention includes (1) histidine-HCl including histidine and histidine monohydrochloride monohydrate, sucrose, methionine, (2) histidine-HCl, sorbitol including histidine and histidine monohydrochloride monohydrate. , Methionine, (3) histidine-HCl including histidine and histidine monohydrochloride monohydrate, trehalose, methionine, (4) histidine-HCl including histidine and histidine monohydrochloride monohydrate, arginine, methionine or (5) ) Histidine and histidine monohydrochloride monohydrate, including histidine-HCl, glycine, and methionine.
하나의 구체예에서, 본 발명에 따른 제형에서 당 또는 당 알코올: 항-Her2 항체의 몰비가 예를 들어, 500:1 내지 700:1일 수 있다.In one embodiment, the molar ratio of sugar or sugar alcohol: anti-Her2 antibody in the formulation according to the present invention may be, for example, 500:1 to 700:1.
상기 몰비는 항-Her2 항체의 분자량을 약 150kDa으로 간주하여 항-Her2 항체의 몰량을 계산하고, 상기 당 또는 당 알코올의 알려진 분자량 (예를 들어, 수크로오스 342.3 g/mol, 솔비톨 182.2 g/mol)을 고려하여, 몰량을 계산한 다음, 항-Her2 항체의 몰량에 대한 당 또는 당 알코올의 몰량의 비율을 계산하여 확인할 수 있다. The molar ratio calculates the molar amount of the anti-Her2 antibody by considering the molecular weight of the anti-Her2 antibody as about 150 kDa, and the known molecular weight of the sugar or sugar alcohol (e.g., sucrose 342.3 g/mol, sorbitol 182.2 g/mol) In consideration of, after calculating the molar amount, the ratio of the molar amount of sugar or sugar alcohol to the molar amount of the anti-Her2 antibody can be calculated and confirmed.
상기 당 또는 당 알코올: 항-Her2 항체의 몰비는 500:1 내지 700:1, 구체적으로 510:1 내지 690:1, 또는 540:1 내지 690:1일 수 있다. 상기 당 또는 당 알코올: 항-Her2 항체의 몰비는 약 500:1, 약 501:1, 약 502:1, 약 503:1, 약 504:1, 약 505:1, 약 506:1, 약 507:1, 약 508:1, 약 509:1, 약 510:1, 약 511:1, 약 512:1, 약 513:1, 약 514:1, 약 515:1, 약 516:1, 약 517:1, 약 518:1, 약 519:1, 약 520:1, 약 521:1, 약 522:1, 약 523:1, 약 524:1, 약 525:1, 약 526:1, 약 527:1, 약 528:1, 약 529:1, 약 530:1, 약 531:1, 약 532:1, 약 533:1, 약 534:1, 약 535:1, 약 536:1, 약 537:1, 약 538:1, 약 539:1, 약 540:1, 약 541:1, 약 542:1, 약 543:1, 약 544:1, 약 545:1, 약 546:1, 약 547:1, 약 548:1, 약 549:1, 약 550:1, 약 551:1, 약 552:1, 약 553:1, 약 554:1, 약 555:1, 약 556:1, 약 557:1, 약 558:1, 약 559:1, 약 560:1, 약 561:1, 약 562:1, 약 563:1, 약 564:1, 약 565:1, 약 566:1, 약 567:1, 약 568:1, 약 569:1, 약 570:1, 약 571:1, 약 572:1, 약 573:1, 약 574:1, 약 575:1, 약 576:1, 약 577:1, 약 578:1, 약 579:1, 약 580:1, 약 581:1, 약 582:1, 약 583:1, 약 584:1, 약 585:1, 약 586:1, 약 587:1, 약 588:1, 약 589:1, 약 590:1, 약 591:1, 약 592:1, 약 593:1, 약 594:1, 약 595:1, 약 596:1, 약 597:1, 약 598:1, 약 599:1, 약 600:1, 약 601:1, 약 602:1, 약 603:1, 약 604:1, 약 605:1, 약 606:1, 약 607:1, 약 608:1, 약 609:1, 약 610:1, 약 611:1, 약 612:1, 약 613:1, 약 614:1, 약 615:1, 약 616:1, 약 617:1, 약 618:1, 약 619:1, 약 620:1, 약 621:1, 약 622:1, 약 623:1, 약 624:1, 약 625:1, 약 626:1, 약 627:1, 약 628:1, 약 629:1, 약 630:1, 약 631:1, 약 632:1, 약 633:1, 약 634:1, 약 635:1, 약 636:1, 약 637:1, 약 638:1, 약 639:1, 약 640:1, 약 641:1, 약 642:1, 약 643:1, 약 644:1, 약 645:1, 약 646:1, 약 647:1, 약 648:1, 약 649:1, 약 650:1, 약 651:1, 약 652:1, 약 653:1, 약 654:1, 약 655:1, 약 656:1, 약 657:1, 약 658:1, 약 659:1, 약 660:1, 약 661:1, 약 662:1, 약 663:1, 약 664:1, 약 665:1, 약 666:1, 약 667:1, 약 668:1, 약 669:1, 약 670:1, 약 671:1, 약 672:1, 약 673:1, 약 674:1, 약 675:1, 약 676:1, 약 677:1, 약 678:1, 약 679:1, 약 680:1, 약 681:1, 약 682:1, 약 683:1, 약 684:1, 약 685:1, 약 686:1, 약 687:1, 약 688:1, 약 689:1, 약 690:1, 약 691:1, 약 692:1, 약 693:1, 약 694:1, 약 695:1, 약 696:1, 약 697:1, 약 698:1, 약 699:1, 또는 약 700:1 (기재된 수치 사이의 값도 포함될 수 있음)일 수 있다. The molar ratio of the sugar or sugar alcohol: anti-Her2 antibody may be 500:1 to 700:1, specifically 510:1 to 690:1, or 540:1 to 690:1. The molar ratio of the sugar or sugar alcohol: anti-Her2 antibody is about 500:1, about 501:1, about 502:1, about 503:1, about 504:1, about 505:1, about 506:1, about 507 :1, about 508:1, about 509:1, about 510:1, about 511:1, about 512:1, about 513:1, about 514:1, about 515:1, about 516:1, about 517 :1, about 518:1, about 519:1, about 520:1, about 521:1, about 522:1, about 523:1, about 524:1, about 525:1, about 526:1, about 527 :1, about 528:1, about 529:1, about 530:1, about 531:1, about 532:1, about 533:1, about 534:1, about 535:1, about 536:1, about 537 :1, about 538:1, about 539:1, about 540:1, about 541:1, about 542:1, about 543:1, about 544:1, about 545:1, about 546:1, about 547 :1, about 548:1, about 549:1, about 550:1, about 551:1, about 552:1, about 553:1, about 554:1, about 555:1, about 556:1, about 557 :1, about 558:1, about 559:1, about 560:1, about 561:1, about 562:1, about 563:1, about 564:1, about 565:1, about 566:1, about 567 :1, about 568:1, about 569:1, about 570:1, about 571:1, about 572:1, about 573:1, about 574:1, about 575:1, about 576:1, about 577 :1, about 578:1, about 579:1, about 580:1, about 581:1, about 582:1, about 583:1, about 584:1, about 585:1, about 586:1, about 587 :1, about 588:1, about 589:1, about 590:1, about 591:1, about 592:1, about 593:1, about 594:1, about 595:1, about 596:1, about 597 :1, about 598:1, about 599:1, about 600:1, about 601:1, about 602:1, about 603:1, about 604:1, about 605:1, about 606:1, about 607 :One , About 608:1, about 609:1, about 610:1, about 611:1, about 612:1, about 613:1, about 614:1, about 615:1, about 616:1, about 617:1 , About 618:1, about 619:1, about 620:1, about 621:1, about 622:1, about 623:1, about 624:1, about 625:1, about 626:1, about 627:1 , About 628:1, about 629:1, about 630:1, about 631:1, about 632:1, about 633:1, about 634:1, about 635:1, about 636:1, about 637:1 , About 638:1, about 639:1, about 640:1, about 641:1, about 642:1, about 643:1, about 644:1, about 645:1, about 646:1, about 647:1 , About 648:1, about 649:1, about 650:1, about 651:1, about 652:1, about 653:1, about 654:1, about 655:1, about 656:1, about 657:1 , About 658:1, about 659:1, about 660:1, about 661:1, about 662:1, about 663:1, about 664:1, about 665:1, about 666:1, about 667:1 , About 668:1, about 669:1, about 670:1, about 671:1, about 672:1, about 673:1, about 674:1, about 675:1, about 676:1, about 677:1 , About 678:1, about 679:1, about 680:1, about 681:1, about 682:1, about 683:1, about 684:1, about 685:1, about 686:1, about 687:1 , About 688:1, about 689:1, about 690:1, about 691:1, about 692:1, about 693:1, about 694:1, about 695:1, about 696:1, about 697:1 , About 698:1, about 699:1, or about 700:1 (values between listed values may also be included).
하나의 구체예에서, 본 발명에 따른 제형은 히스티딘과 이의 염 예를 들어, 히스티딘 : 히스티딘 모노하이드로클로라이드 일수화물을 0.2 내지 3.0:1의 몰비로 포함할 수 있다. 히스티딘 : 히스티딘 모노하이드로클로라이드 일수화물을 약 0.2, 약 0.3, 약 0.4, 약 0.5, 약 0.6, 약 0.7, 약 0.8, 약 0.9, 약 1.0, 약 1.1, 약 1.2, 약 1.3, 약 1.4, 약 1.5, 약 1.6, 약 1.7, 약 1.8, 약 1.9, 약 2.0, 약 2.1, 약 2.2, 약 2.3, 약 2.4, 약 2.5, 약 2.6, 약 2.7, 약 2.8, 약 2.9, 약 3.0 (기재된 수치 사이의 값도 포함될 수 있음)의 몰비로 포함할 수 있다. In one embodiment, the formulation according to the present invention may comprise histidine and a salt thereof, for example, histidine: histidine monohydrochloride monohydrate in a molar ratio of 0.2 to 3.0:1. Histidine: Histidine monohydrochloride monohydrate about 0.2, about 0.3, about 0.4, about 0.5, about 0.6, about 0.7, about 0.8, about 0.9, about 1.0, about 1.1, about 1.2, about 1.3, about 1.4, about 1.5 , About 1.6, about 1.7, about 1.8, about 1.9, about 2.0, about 2.1, about 2.2, about 2.3, about 2.4, about 2.5, about 2.6, about 2.7, about 2.8, about 2.9, about 3.0 Value may also be included).
몰비는 히스티딘 예를 들어 L-히스티딘의 알려진 분자량 (예를 들어, 히스티딘 155.2 g/mol)을 고려하여 히스티딘의 몰량을 계산하고, 히스티딘 모노하이드로클로라이드 일수화물의 알려진 분자량 (예를 들어, 209.6 g/mol)을 고려하여 몰량을 계산한 다음, 히스티딘의 몰량에 대하여, 히스티딘 모노하이드로클로라이드 일수화물 몰량의 비율을 계산하여 확인할 수 있다. The molar ratio calculates the molar amount of histidine taking into account the known molecular weight of histidine, e.g. L-histidine (e.g., 155.2 g/mol histidine), and the known molecular weight of histidine monohydrochloride monohydrate (e.g., 209.6 g/mol) mol), the molar amount is calculated, and then the ratio of the molar amount of histidine monohydrochloride monohydrate to the molar amount of histidine can be calculated and confirmed.
본 발명에 따른 제형은 5 mM 내지 50 mM, 예를 들어 10 mM 내지 40 mM, 15 mM 내지 35 mM, 또는 20 mM 내지 30 mM 농도의 히스티딘을 포함할 수 있다. 상기 히스티딘은 약 5 mM, 약 6 mM, 약 7 mM, 약 8 mM, 약 9 mM, 약 10 mM, 약 11 mM, 약 12 mM, 약 13 mM, 약 14 mM, 약 15 mM, 약 16 mM, 약 17 mM, 약 18 mM, 약 19 mM, 약 20 mM, 약 21 mM, 약 22 mM, 약 23 mM, 약 24 mM, 약 25 mM, 약 26 mM, 약 27 mM, 약 28 mM, 약 29 mM, 약 30 mM, 약 31 mM, 약 32 mM, 약 33 mM, 약 34 mM, 약 35 mM, 약 36 mM, 약 37 mM, 약 38 mM, 약 39 mM, 약 40 mM, 약 41 mM, 약 42 mM, 약 43 mM, 약 44 mM, 약 45 mM, 약 46 mM, 약 47 mM, 약 48 mM, 약 49 mM, 또는 약 50 mM (기재된 수치 사이의 값도 포함될 수 있음)의 농도로 포함될 수 있다. The formulation according to the present invention may contain histidine at a concentration of 5 mM to 50 mM, for example 10 mM to 40 mM, 15 mM to 35 mM, or 20 mM to 30 mM. The histidine is about 5 mM, about 6 mM, about 7 mM, about 8 mM, about 9 mM, about 10 mM, about 11 mM, about 12 mM, about 13 mM, about 14 mM, about 15 mM, about 16 mM , About 17 mM, about 18 mM, about 19 mM, about 20 mM, about 21 mM, about 22 mM, about 23 mM, about 24 mM, about 25 mM, about 26 mM, about 27 mM, about 28 mM, about 29 mM, about 30 mM, about 31 mM, about 32 mM, about 33 mM, about 34 mM, about 35 mM, about 36 mM, about 37 mM, about 38 mM, about 39 mM, about 40 mM, about 41 mM , About 42 mM, about 43 mM, about 44 mM, about 45 mM, about 46 mM, about 47 mM, about 48 mM, about 49 mM, or about 50 mM (values between stated values may also be included). Can be included as.
상기 제형은 예를 들어, 메티오닌, 시스테인, 알지닌, 글리신, 프롤린, 라이신, 글루탐산, 이소루이신, 페닐알라닌, 티로신, 아스파트산으로 구성된 군에서 선택되는 히스티딘 이외의 아미노산을 5 mM 내지 30 mM의 농도로 포함할 수 있다. 상기 히스티딘 이외의 아미노산은 5 mM 내지 30 mM, 예를 들어 7 mM 내지 20 mM, 또는 9 mM 내지 15 mM의 농도로 포함될 수 있다. 구체적으로, 약 5 mM, 약 6 mM, 약 7 mM, 약 8 mM, 약 9 mM, 약 10 mM, 약 11 mM, 약 12 mM, 약 13 mM, 약 14 mM, 약 15 mM, 약 16 mM, 약 17 mM, 약 18 mM, 약 19 mM, 약 20 mM, 약 21 mM, 약 22 mM, 약 23 mM, 약 24 mM, 약 25 mM, 약 26 mM, 약 27 mM, 약 28 mM, 약 29 mM, 또는 약 30 mM (기재된 수치 사이의 값도 포함될 수 있음)의 농도로 포함될 수 있다.The formulation includes, for example, 5 mM to 30 mM of amino acids other than histidine selected from the group consisting of methionine, cysteine, arginine, glycine, proline, lysine, glutamic acid, isoleucine, phenylalanine, tyrosine, and aspartic acid. It can be included in concentration. Amino acids other than histidine may be included in a concentration of 5 mM to 30 mM, for example, 7 mM to 20 mM, or 9 mM to 15 mM. Specifically, about 5 mM, about 6 mM, about 7 mM, about 8 mM, about 9 mM, about 10 mM, about 11 mM, about 12 mM, about 13 mM, about 14 mM, about 15 mM, about 16 mM , About 17 mM, about 18 mM, about 19 mM, about 20 mM, about 21 mM, about 22 mM, about 23 mM, about 24 mM, about 25 mM, about 26 mM, about 27 mM, about 28 mM, about 29 mM, or about 30 mM (values between stated values may also be included).
경우에 따라서, 상기 제형은 예를 들어, 메티오닌, 시스테인, 알지닌, 글리신, 프롤린, 라이신, 글루탐산, 이소루이신, 페닐알라닌, 티로신, 아스파트산으로 구성된 군에서 선택되는 히스티딘 이외의 아미노산을 50 mM 내지 150 mM의 농도로 포함할 수 있다. 구체적으로, 상기 히스티딘 이외의 아미노산은 50 mM 내지 150 mM, 60 mM 내지 140 mM, 70 mM 내지 135 mM, 80 mM 내지 135 mM, 90 mM 내지 135 mM, 100 mM 내지 140 mM 또는 100 mM 내지 135 mM의 농도로 포함될 수 있다. 구체적으로, 상기 히스티딘 이외의 아미노산은 약 50 mM, 약 51 mM, 약 52 mM, 약 53 mM, 약 54 mM, 약 55 mM, 약 55 mM, 약 56 mM, 약 57 mM, 약 58 mM, 약 59 mM, 약 60 mM, 약 61 mM, 약 62 mM, 약 63 mM, 약 64 mM, 약 65 mM, 약 66 mM, 약 67 mM, 약 68 mM, 약 69 mM, 약 70 mM, 약 71 mM, 약 72 mM, 약 73 mM, 약 74 mM, 약 75 mM, 약 76 mM, 약 77 mM, 약 78 mM, 약 79 mM, 약 80 mM, 약 81 mM, 약 82 mM, 약 83 mM, 약 84 mM, 약 85 mM, 약 86 mM, 약 87 mM, 약 88 mM, 약 89 mM, 약 90 mM, 약 91 mM, 약 92 mM, 약 93 mM, 약 94 mM, 약 95 mM, 약 96 mM, 약 97 mM, 약 98 mM, 약 99 mM, 약 100 mM, 약 101 mM, 약 102 mM, 약 103 mM, 약 104 mM, 약 105 mM, 약 106 mM, 약 107 mM, 약 108 mM, 약 109 mM, 약 110 mM, 약 111 mM, 약 112 mM, 약 113 mM, 약 114 mM, 약 115 mM, 약 116 mM, 약 117 mM, 약 118 mM, 약 119 mM, 약 120 mM, 약 121 mM, 약 122 mM, 약 123 mM, 약 124 mM, 약 125 mM, 약 126 mM, 약 127 mM, 약 128 mM, 약 129 mM, 약 130 mM, 약 131 mM, 약 132 mM, 약 133 mM, 약 134 mM, 약 135 mM, 약 136 mM, 약 137 mM, 약 138 mM, 약 139 mM, 약 140 mM, 약 141 mM, 약 142 mM, 약 143 mM, 약 144 mM, 약 145 mM, 약 146 mM, 약 147 mM, 약 148 mM, 약 149 mM, 또는 약 150 mM (기재된 수치 사이의 값도 포함될 수 있음)의 농도로 포함될 수 있다.In some cases, the formulation contains 50 mM amino acids other than histidine selected from the group consisting of, for example, methionine, cysteine, arginine, glycine, proline, lysine, glutamic acid, isoleucine, phenylalanine, tyrosine, and aspartic acid. To 150 mM. Specifically, amino acids other than histidine are 50 mM to 150 mM, 60 mM to 140 mM, 70 mM to 135 mM, 80 mM to 135 mM, 90 mM to 135 mM, 100 mM to 140 mM, or 100 mM to 135 mM. It can be included in the concentration of. Specifically, the amino acid other than histidine is about 50 mM, about 51 mM, about 52 mM, about 53 mM, about 54 mM, about 55 mM, about 55 mM, about 56 mM, about 57 mM, about 58 mM, about 59 mM, about 60 mM, about 61 mM, about 62 mM, about 63 mM, about 64 mM, about 65 mM, about 66 mM, about 67 mM, about 68 mM, about 69 mM, about 70 mM, about 71 mM , About 72 mM, about 73 mM, about 74 mM, about 75 mM, about 76 mM, about 77 mM, about 78 mM, about 79 mM, about 80 mM, about 81 mM, about 82 mM, about 83 mM, about 84 mM, about 85 mM, about 86 mM, about 87 mM, about 88 mM, about 89 mM, about 90 mM, about 91 mM, about 92 mM, about 93 mM, about 94 mM, about 95 mM, about 96 mM , About 97 mM, about 98 mM, about 99 mM, about 100 mM, about 101 mM, about 102 mM, about 103 mM, about 104 mM, about 105 mM, about 106 mM, about 107 mM, about 108 mM, about 109 mM, about 110 mM, about 111 mM, about 112 mM, about 113 mM, about 114 mM, about 115 mM, about 116 mM, about 117 mM, about 118 mM, about 119 mM, about 120 mM, about 121 mM , About 122 mM, about 123 mM, about 124 mM, about 125 mM, about 126 mM, about 127 mM, about 128 mM, about 129 mM, about 130 mM, about 131 mM, about 132 mM, about 133 mM, about 134 mM, about 135 mM, about 136 mM, about 137 mM, about 138 mM, about 139 mM, about 140 mM, about 141 mM, about 142 mM, about 143 mM, about 144 mM, about 145 mM, about 146 mM , About 147 mM, about 148 mM, about 149 mM, or about 150 mM (described Values between values may also be included).
상기 항-Her2 항체: 히스티딘 이외의 아미노산의 몰비는 1:10 내지 1:100, 예를 들어 1:20 내지 1:80, 또는 1:30 내지 1:70일 수 있다. 구체적으로, 상기 항-Her2 항체: 히스티딘 이외의 아미노산의 몰비는 약 1:10, 약 1:11, 약 1:12, 약 1:13, 약 1:14, 약 1:15, 약 1:16, 약 1:17, 약 1:18, 약 1:19, 약 1:20, 약 1:21, 약 1:22, 약 1:23, 약 1:24, 약 1:25, 약 1:26, 약 1:27, 약 1:28, 약 1:29, 약 1:30, 약 1:31, 약 1:32, 약 1:33, 약 1:34, 약 1:35, 약 1:36, 약 1:37, 약 1:38, 약 1:39, 약 1:40, 약 1:41, 약 1:42, 약 1:43, 약 1:44, 약 1:45, 약 1:46, 약 1:47, 약 1:48, 약 1:49, 약 1:50, 약 1:51, 약 1:52, 약 1:53, 약 1:54, 약 1:55, 약 1:56, 약 1:57, 약 1:58, 약 1:59, 약 1:60, 약 1:61, 약 1:62, 약 1:63, 약 1:64, 약 1:65, 약 1:66, 약 1:67, 약 1:68, 약 1:69, 약 1:70, 약 1:71, 약 1:72, 약 1:73, 약 1:74, 약 1:75, 약 1:76, 약 1:77, 약 1:78, 약 1:79, 약 1:80, 약 1:81, 약 1:82, 약 1:83, 약 1:84, 약 1:85, 약 1:86, 약 1:87, 약 1:88, 약 1:89, 약 1:90, 약 1:91, 약 1:92, 약 1:93, 약 1:94, 약 1:95, 약 1:96, 약 1:97, 약 1:98, 약 1:99, 약 1:100 (기재된 수치 사이의 값도 포함될 수 있음)일 수 있다.The molar ratio of the anti-Her2 antibody: amino acids other than histidine may be 1:10 to 1:100, for example, 1:20 to 1:80, or 1:30 to 1:70. Specifically, the molar ratio of the anti-Her2 antibody: amino acids other than histidine is about 1:10, about 1:11, about 1:12, about 1:13, about 1:14, about 1:15, about 1:16 , About 1:17, about 1:18, about 1:19, about 1:20, about 1:21, about 1:22, about 1:23, about 1:24, about 1:25, about 1:26 , About 1:27, about 1:28, about 1:29, about 1:30, about 1:31, about 1:32, about 1:33, about 1:34, about 1:35, about 1:36 , About 1:37, about 1:38, about 1:39, about 1:40, about 1:41, about 1:42, about 1:43, about 1:44, about 1:45, about 1:46 , About 1:47, about 1:48, about 1:49, about 1:50, about 1:51, about 1:52, about 1:53, about 1:54, about 1:55, about 1:56 , About 1:57, about 1:58, about 1:59, about 1:60, about 1:61, about 1:62, about 1:63, about 1:64, about 1:65, about 1:66 , About 1:67, about 1:68, about 1:69, about 1:70, about 1:71, about 1:72, about 1:73, about 1:74, about 1:75, about 1:76 , About 1:77, about 1:78, about 1:79, about 1:80, about 1:81, about 1:82, about 1:83, about 1:84, about 1:85, about 1:86 , About 1:87, about 1:88, about 1:89, about 1:90, about 1:91, about 1:92, about 1:93, about 1:94, about 1:95, about 1:96 , About 1:97, about 1:98, about 1:99, about 1:100 (values between the listed values may also be included).
경우에 따라서, 상기 히스티딘 이외의 아미노산: 항-Her2 항체의 몰비는 300 : 1 내지 450:1, 구체적으로 330:1 내지 420:1, 또는 350:1 내지 400:1일 수 있다. 상기 히스티딘 이외의 아미노산: 항-Her2 항체의 몰비는 약 300 : 1, 약 301:1, 약 302:1, 약 303:1, 약 304:1, 약 305:1, 약 306:1, 약 307:1, 약 308:1, 약 309:1, 약 310:1, 약 311:1, 약 312:1, 약 313:1, 약 314:1, 약 315:1, 약 316:1, 약 317:1, 약 318:1, 약 319:1, 약 320:1, 약 321:1, 약 322:1, 약 323:1, 약 324:1, 약 325:1, 약 326:1, 약 327:1, 약 328:1, 약 329:1, 약 330:1, 약 331:1, 약 332:1, 약 333:1, 약 334:1, 약 335:1, 약 336:1, 약 337:1, 약 338:1, 약 339:1, 약 340:1, 약 341:1, 약 342:1, 약 343:1, 약 344:1, 약 345:1, 약 346:1, 약 347:1, 약 348:1, 약 349:1, 약 350:1, 약 351:1, 약 352:1, 약 353:1, 약 354:1, 약 355:1, 약 356:1, 약 357:1, 약 358:1, 약 359:1, 약 360:1, 약 361:1, 약 362:1, 약 363:1, 약 364:1, 약 365:1, 약 366:1, 약 367:1, 약 368:1, 약 369:1, 약 370:1, 약 371:1, 약 372:1, 약 373:1, 약 374:1, 약 375:1, 약 376:1, 약 377:1, 약 378:1, 약 379:1, 약 380:1, 약 381:1, 약 382:1, 약 383:1, 약 384:1, 약 385:1, 약 386:1, 약 387:1, 약 388:1, 약 389:1, 약 390:1, 약 391:1, 약 392:1, 약 393:1, 약 394:1, 약 395:1, 약 396:1, 약 397:1, 약 398:1, 약 399:1, 약 400:1, 약 401:1, 약 402:1, 약 403:1, 약 404:1, 약 405:1, 약 406:1, 약 407:1, 약 408:1, 약 409:1, 약 410:1, 약 411:1, 약 412:1, 약 413:1, 약 414:1, 약 415:1, 약 416:1, 약 417:1, 약 418:1, 약 419:1, 약 420:1, 약 421:1, 약 422:1, 약 423:1, 약 424:1, 약 425:1, 약 426:1, 약 427:1, 약 428:1, 약 429:1, 약 430:1, 약 431:1, 약 432:1, 약 433:1, 약 434:1, 약 435:1, 약 436:1, 약 437:1, 약 438:1, 약 439:1, 약 440:1, 약 441:1, 약 442:1, 약 443:1, 약 444:1, 약 445:1, 약 446:1, 약 447:1, 약 448:1, 약 449:1, 약 450: 1 (기재된 수치 사이의 값도 포함될 수 있음)일 수 있다.In some cases, the molar ratio of amino acids other than histidine: anti-Her2 antibody may be 300:1 to 450:1, specifically 330:1 to 420:1, or 350:1 to 400:1. The molar ratio of amino acids other than histidine: anti-Her2 antibody is about 300:1, about 301:1, about 302:1, about 303:1, about 304:1, about 305:1, about 306:1, about 307 :1, about 308:1, about 309:1, about 310:1, about 311:1, about 312:1, about 313:1, about 314:1, about 315:1, about 316:1, about 317 :1, about 318:1, about 319:1, about 320:1, about 321:1, about 322:1, about 323:1, about 324:1, about 325:1, about 326:1, about 327 :1, about 328:1, about 329:1, about 330:1, about 331:1, about 332:1, about 333:1, about 334:1, about 335:1, about 336:1, about 337 :1, about 338:1, about 339:1, about 340:1, about 341:1, about 342:1, about 343:1, about 344:1, about 345:1, about 346:1, about 347 :1, about 348:1, about 349:1, about 350:1, about 351:1, about 352:1, about 353:1, about 354:1, about 355:1, about 356:1, about 357 :1, about 358:1, about 359:1, about 360:1, about 361:1, about 362:1, about 363:1, about 364:1, about 365:1, about 366:1, about 367 :1, about 368:1, about 369:1, about 370:1, about 371:1, about 372:1, about 373:1, about 374:1, about 375:1, about 376:1, about 377 :1, about 378:1, about 379:1, about 380:1, about 381:1, about 382:1, about 383:1, about 384:1, about 385:1, about 386:1, about 387 :1, about 388:1, about 389:1, about 390:1, about 391:1, about 392:1, about 393:1, about 394:1, about 395:1, about 396:1, about 397 :1, about 398:1, about 399:1, about 400:1, about 401:1, about 402:1, about 403:1, about 404:1, about 405:1, about 406:1, about 407:1, about 408:1, about 409:1, about 410:1, about 411:1, about 412:1, about 413:1, about 414:1, about 415:1, about 416:1, about 417:1, about 418:1, about 419:1, about 420:1, about 421:1, about 422:1, about 423:1, about 424:1, about 425:1, about 426:1, about 427:1, about 428:1, about 429:1, about 430:1, about 431:1, about 432:1, about 433:1, about 434:1, about 435:1, about 436:1, about 437:1, about 438:1, about 439:1, about 440:1, about 441:1, about 442:1, about 443:1, about 444:1, about 445:1, about 446:1, about It can be 447:1, about 448:1, about 449:1, about 450:1 (values between the listed values may also be included).
경우에 따라서, 상기 히스티딘 이외의 아미노산: 항-Her2 항체의 몰비는 500:1 내지 700:1, 구체적으로 510:1 내지 690:1, 또는 540:1 내지 690:1일 수 있다. 상기 히스티딘 이외의 아미노산: 항-Her2 항체의 몰비는 약 500:1, 약 501:1, 약 502:1, 약 503:1, 약 504:1, 약 505:1, 약 506:1, 약 507:1, 약 508:1, 약 509:1, 약 510:1, 약 511:1, 약 512:1, 약 513:1, 약 514:1, 약 515:1, 약 516:1, 약 517:1, 약 518:1, 약 519:1, 약 520:1, 약 521:1, 약 522:1, 약 523:1, 약 524:1, 약 525:1, 약 526:1, 약 527:1, 약 528:1, 약 529:1, 약 530:1, 약 531:1, 약 532:1, 약 533:1, 약 534:1, 약 535:1, 약 536:1, 약 537:1, 약 538:1, 약 539:1, 약 540:1, 약 541:1, 약 542:1, 약 543:1, 약 544:1, 약 545:1, 약 546:1, 약 547:1, 약 548:1, 약 549:1, 약 550:1, 약 551:1, 약 552:1, 약 553:1, 약 554:1, 약 555:1, 약 556:1, 약 557:1, 약 558:1, 약 559:1, 약 560:1, 약 561:1, 약 562:1, 약 563:1, 약 564:1, 약 565:1, 약 566:1, 약 567:1, 약 568:1, 약 569:1, 약 570:1, 약 571:1, 약 572:1, 약 573:1, 약 574:1, 약 575:1, 약 576:1, 약 577:1, 약 578:1, 약 579:1, 약 580:1, 약 581:1, 약 582:1, 약 583:1, 약 584:1, 약 585:1, 약 586:1, 약 587:1, 약 588:1, 약 589:1, 약 590:1, 약 591:1, 약 592:1, 약 593:1, 약 594:1, 약 595:1, 약 596:1, 약 597:1, 약 598:1, 약 599:1, 약 600:1, 약 601:1, 약 602:1, 약 603:1, 약 604:1, 약 605:1, 약 606:1, 약 607:1, 약 608:1, 약 609:1, 약 610:1, 약 611:1, 약 612:1, 약 613:1, 약 614:1, 약 615:1, 약 616:1, 약 617:1, 약 618:1, 약 619:1, 약 620:1, 약 621:1, 약 622:1, 약 623:1, 약 624:1, 약 625:1, 약 626:1, 약 627:1, 약 628:1, 약 629:1, 약 630:1, 약 631:1, 약 632:1, 약 633:1, 약 634:1, 약 635:1, 약 636:1, 약 637:1, 약 638:1, 약 639:1, 약 640:1, 약 641:1, 약 642:1, 약 643:1, 약 644:1, 약 645:1, 약 646:1, 약 647:1, 약 648:1, 약 649:1, 약 650:1, 약 651:1, 약 652:1, 약 653:1, 약 654:1, 약 655:1, 약 656:1, 약 657:1, 약 658:1, 약 659:1, 약 660:1, 약 661:1, 약 662:1, 약 663:1, 약 664:1, 약 665:1, 약 666:1, 약 667:1, 약 668:1, 약 669:1, 약 670:1, 약 671:1, 약 672:1, 약 673:1, 약 674:1, 약 675:1, 약 676:1, 약 677:1, 약 678:1, 약 679:1, 약 680:1, 약 681:1, 약 682:1, 약 683:1, 약 684:1, 약 685:1, 약 686:1, 약 687:1, 약 688:1, 약 689:1, 약 690:1, 약 691:1, 약 692:1, 약 693:1, 약 694:1, 약 695:1, 약 696:1, 약 697:1, 약 698:1, 약 699:1, 또는 약 700:1 (기재된 수치 사이의 값도 포함될 수 있음)일 수 있다. In some cases, the molar ratio of amino acids other than histidine: anti-Her2 antibody may be 500:1 to 700:1, specifically 510:1 to 690:1, or 540:1 to 690:1. The molar ratio of amino acids other than histidine: anti-Her2 antibody is about 500:1, about 501:1, about 502:1, about 503:1, about 504:1, about 505:1, about 506:1, about 507 :1, about 508:1, about 509:1, about 510:1, about 511:1, about 512:1, about 513:1, about 514:1, about 515:1, about 516:1, about 517 :1, about 518:1, about 519:1, about 520:1, about 521:1, about 522:1, about 523:1, about 524:1, about 525:1, about 526:1, about 527 :1, about 528:1, about 529:1, about 530:1, about 531:1, about 532:1, about 533:1, about 534:1, about 535:1, about 536:1, about 537 :1, about 538:1, about 539:1, about 540:1, about 541:1, about 542:1, about 543:1, about 544:1, about 545:1, about 546:1, about 547 :1, about 548:1, about 549:1, about 550:1, about 551:1, about 552:1, about 553:1, about 554:1, about 555:1, about 556:1, about 557 :1, about 558:1, about 559:1, about 560:1, about 561:1, about 562:1, about 563:1, about 564:1, about 565:1, about 566:1, about 567 :1, about 568:1, about 569:1, about 570:1, about 571:1, about 572:1, about 573:1, about 574:1, about 575:1, about 576:1, about 577 :1, about 578:1, about 579:1, about 580:1, about 581:1, about 582:1, about 583:1, about 584:1, about 585:1, about 586:1, about 587 :1, about 588:1, about 589:1, about 590:1, about 591:1, about 592:1, about 593:1, about 594:1, about 595:1, about 596:1, about 597 :1, about 598:1, about 599:1, about 600:1, about 601:1, about 602:1, about 603:1, about 604:1, about 605:1, about 606:1, about 60 7:1, about 608:1, about 609:1, about 610:1, about 611:1, about 612:1, about 613:1, about 614:1, about 615:1, about 616:1, about 617:1, about 618:1, about 619:1, about 620:1, about 621:1, about 622:1, about 623:1, about 624:1, about 625:1, about 626:1, about 627:1, about 628:1, about 629:1, about 630:1, about 631:1, about 632:1, about 633:1, about 634:1, about 635:1, about 636:1, about 637:1, about 638:1, about 639:1, about 640:1, about 641:1, about 642:1, about 643:1, about 644:1, about 645:1, about 646:1, about 647:1, about 648:1, about 649:1, about 650:1, about 651:1, about 652:1, about 653:1, about 654:1, about 655:1, about 656:1, about 657:1, about 658:1, about 659:1, about 660:1, about 661:1, about 662:1, about 663:1, about 664:1, about 665:1, about 666:1, about 667:1, about 668:1, about 669:1, about 670:1, about 671:1, about 672:1, about 673:1, about 674:1, about 675:1, about 676:1, about 677:1, about 678:1, about 679:1, about 680:1, about 681:1, about 682:1, about 683:1, about 684:1, about 685:1, about 686:1, about 687:1, about 688:1, about 689:1, about 690:1, about 691:1, about 692:1, about 693:1, about 694:1, about 695:1, about 696:1, about It may be 697:1, about 698:1, about 699:1, or about 700:1 (values between listed values may also be included).
몰비에 대한 계산은 앞서 설명한 바와 동일하다. 항-Her2 항체의 분자량을 약 150kDa으로 간주하여 항-Her2 항체의 몰량을 계산하고, 히스티딘 이외의 아미노산의 알려진 분자량을 고려하여 몰량을 계산한 다음, 항-Her2 항체의 몰량에 대하여 히스티딘 이외의 아미노산 몰량의 비율을 계산하여 확인할 수 있다. The calculation for the molar ratio is the same as described above. The molar amount of the anti-Her2 antibody is calculated by considering the molecular weight of the anti-Her2 antibody as about 150 kDa, and the molar amount is calculated by considering the known molecular weight of amino acids other than histidine, and then amino acids other than histidine are calculated based on the molar amount of the anti-Her2 antibody. It can be checked by calculating the ratio of the molar amount.
상기 제형은 50 mM 내지 150 mM의 당 또는 당 알코올을 포함할 수 있다. 구체적으로, 상기 당 또는 당 알코올은 50 mM 내지 150 mM, 60 mM 내지 140 mM, 70 mM 내지 135 mM, 80 mM 내지 135 mM, 90 mM 내지 135 mM, 100 mM 내지 140 mM 또는 100 mM 내지 135 mM의 농도로 포함될 수 있다. 구체적으로, 상기 당 또는 당 알코올은 약 50 mM, 약 51 mM, 약 52 mM, 약 53 mM, 약 54 mM, 약 55 mM, 약 55 mM, 약 56 mM, 약 57 mM, 약 58 mM, 약 59 mM, 약 60 mM, 약 61 mM, 약 62 mM, 약 63 mM, 약 64 mM, 약 65 mM, 약 66 mM, 약 67 mM, 약 68 mM, 약 69 mM, 약 70 mM, 약 71 mM, 약 72 mM, 약 73 mM, 약 74 mM, 약 75 mM, 약 76 mM, 약 77 mM, 약 78 mM, 약 79 mM, 약 80 mM, 약 81 mM, 약 82 mM, 약 83 mM, 약 84 mM, 약 85 mM, 약 86 mM, 약 87 mM, 약 88 mM, 약 89 mM, 약 90 mM, 약 91 mM, 약 92 mM, 약 93 mM, 약 94 mM, 약 95 mM, 약 96 mM, 약 97 mM, 약 98 mM, 약 99 mM, 약 100 mM, 약 101 mM, 약 102 mM, 약 103 mM, 약 104 mM, 약 105 mM, 약 106 mM, 약 107 mM, 약 108 mM, 약 109 mM, 약 110 mM, 약 111 mM, 약 112 mM, 약 113 mM, 약 114 mM, 약 115 mM, 약 116 mM, 약 117 mM, 약 118 mM, 약 119 mM, 약 120 mM, 약 121 mM, 약 122 mM, 약 123 mM, 약 124 mM, 약 125 mM, 약 126 mM, 약 127 mM, 약 128 mM, 약 129 mM, 약 130 mM, 약 131 mM, 약 132 mM, 약 133 mM, 약 134 mM, 약 135 mM, 약 136 mM, 약 137 mM, 약 138 mM, 약 139 mM, 약 140 mM, 약 141 mM, 약 142 mM, 약 143 mM, 약 144 mM, 약 145 mM, 약 146 mM, 약 147 mM, 약 148 mM, 약 149 mM, 또는 약 150 mM (기재된 수치 사이의 값도 포함될 수 있음)의 농도로 포함될 수 있다.The formulation may contain 50 mM to 150 mM sugar or sugar alcohol. Specifically, the sugar or sugar alcohol is 50 mM to 150 mM, 60 mM to 140 mM, 70 mM to 135 mM, 80 mM to 135 mM, 90 mM to 135 mM, 100 mM to 140 mM, or 100 mM to 135 mM It can be included in the concentration of. Specifically, the sugar or sugar alcohol is about 50 mM, about 51 mM, about 52 mM, about 53 mM, about 54 mM, about 55 mM, about 55 mM, about 56 mM, about 57 mM, about 58 mM, about 59 mM, about 60 mM, about 61 mM, about 62 mM, about 63 mM, about 64 mM, about 65 mM, about 66 mM, about 67 mM, about 68 mM, about 69 mM, about 70 mM, about 71 mM , About 72 mM, about 73 mM, about 74 mM, about 75 mM, about 76 mM, about 77 mM, about 78 mM, about 79 mM, about 80 mM, about 81 mM, about 82 mM, about 83 mM, about 84 mM, about 85 mM, about 86 mM, about 87 mM, about 88 mM, about 89 mM, about 90 mM, about 91 mM, about 92 mM, about 93 mM, about 94 mM, about 95 mM, about 96 mM , About 97 mM, about 98 mM, about 99 mM, about 100 mM, about 101 mM, about 102 mM, about 103 mM, about 104 mM, about 105 mM, about 106 mM, about 107 mM, about 108 mM, about 109 mM, about 110 mM, about 111 mM, about 112 mM, about 113 mM, about 114 mM, about 115 mM, about 116 mM, about 117 mM, about 118 mM, about 119 mM, about 120 mM, about 121 mM , About 122 mM, about 123 mM, about 124 mM, about 125 mM, about 126 mM, about 127 mM, about 128 mM, about 129 mM, about 130 mM, about 131 mM, about 132 mM, about 133 mM, about 134 mM, about 135 mM, about 136 mM, about 137 mM, about 138 mM, about 139 mM, about 140 mM, about 141 mM, about 142 mM, about 143 mM, about 144 mM, about 145 mM, about 146 mM , About 147 mM, about 148 mM, about 149 mM, or about 150 mM A value between may also be included).
하나의 구체예에서, i) 5 내지 50 mM 히스티딘-HCl, ii) 50 mM 내지 150 mM 수크로오스, 솔비톨, 트레할로스, 알지닌 또는 글리신 및 선택적으로 iii) 5 mM 내지 30 mM 메티오닌을 포함할 수 있다. 앞서 설명한 바와 같이, 5 내지 50 mM, 50 mM 내지 150 mM 및 5 mM 내지 30 mM의 농도 범위로 기재된 경우, 본 발명의 제형은 각 범위 내에서 임의의 농도를 모두 포함하는 것으로 이해할 수 있다. In one embodiment, i) 5 to 50 mM histidine-HCl, ii) 50 mM to 150 mM sucrose, sorbitol, trehalose, arginine or glycine and optionally iii) 5 mM to 30 mM methionine. As described above, when described in concentration ranges of 5 to 50 mM, 50 mM to 150 mM, and 5 mM to 30 mM, it can be understood that the formulation of the present invention includes all of any concentrations within each range.
경우에 따라서, 상기 제형은 팽화제(bulking agent), 점도 조절제, 안정화제, 계면활성제, 항산화제 또는 등장화제(tonicity agent) 등을 추가로 포함할 수 있다. In some cases, the formulation may further include a bulking agent, a viscosity modifier, a stabilizer, a surfactant, an antioxidant, or a tonicity agent.
본 발명에 따른 제형은 예를 들어, 폴리솔베이트, 폴록사머, SDS (sodium dodecyl sulfate), SLS (sodium laurel sulfate) 등을 포함할 수 있다. 상기 폴리솔베이트는 폴리솔베이트 20, 폴리솔베이트 21, 폴리솔베이트 40, 폴리솔베이트 60, 폴리솔베이트 61, 폴리솔베이트 65, 폴리솔베이트 80, 폴리솔베이트 81, 폴리솔베이트 85 또는 이의 혼합물을 포함할 수 있다. 구체적으로, 본 발명에 따른 제형은 폴리솔베이트, 예를 들어 폴리솔베이트 20을 포함할 수 있다.The formulation according to the present invention may include, for example, polysorbate, poloxamer, sodium dodecyl sulfate (SDS), sodium laurel sulfate (SLS), and the like. The polysorbate is polysorbate 20, polysorbate 21, polysorbate 40, polysorbate 60, polysorbate 61, polysorbate 65, polysorbate 80, polysorbate 81, polysorbate 85 or its It may contain a mixture. Specifically, the formulation according to the present invention may comprise a polysorbate, for example polysorbate 20.
본 발명에 따른 제형은 0.01 내지 0.1% (w/v) 또는 0.02 내지 0.08% (w/v)의 농도로 폴리솔베이트를 포함할 수 있다. 본 발명에 따른 제형은 약 0.01% (w/v), 약 0.02% (w/v), 약 0.03% (w/v), 약 0.04% (w/v), 약 0.05% (w/v), 약 0.06% (w/v), 약 0.07% (w/v) 또는 약 0.08% (w/v) (기재된 수치 사이의 값도 포함될 수 있음)의 농도로 폴리솔베이트를 포함할 수 있다.The formulation according to the present invention may comprise a polysorbate in a concentration of 0.01 to 0.1% (w/v) or 0.02 to 0.08% (w/v). Formulations according to the invention are about 0.01% (w/v), about 0.02% (w/v), about 0.03% (w/v), about 0.04% (w/v), about 0.05% (w/v) , About 0.06% (w/v), about 0.07% (w/v), or about 0.08% (w/v) (values between listed values may also be included).
상기 항-Her2 항체는 예를 들어, 퍼투주맙(pertuzumab)일 수 있다. 본 명세서의 퍼투주맙은 HER2를 표적으로 하여 Perjeta®로 시판되고 있는 단일클론항체이다. 퍼투주맙은 HER2 수용체의 세포외기질 도메인의 영역 II에 결합하여 이량체의 형성을 억제하고, 이를 통해 수용체-매개성 신호경로를 저해한다. HER2/neu로도 알려진 ErbB2는 EGFR family의 두 번째 구성원으로, EGFR 패밀리 내 다른 세 개 EGFR/HER1, HER3 및 HER4의 구성원과 헤테로다이머를 형성함으로써, 생물학적 기능을 나타낸다. ErbB2를 암호화화는 neu 유전자는 래트 신경아세포종(neuroblastoma)에서 최초로 분리되었다. HER2로도 알려진 인간 체세포 내 neu 유전자의 상동 유전자는 염색체 17q21.1상에 위치한다. 암호화된 생성물은 ErbB2이며, 대략 185kDa의 분자량을 갖는 1255개의 아미노산으로 구성되어 있으며, 720 내지 987번째 아미노산 위치는 티로신 키나아제 활성 도메인에 속한다. ErbB2는 PI3K 및 MAPK 신호전달경로에 의존적이고, 사이클린 D 및 c-myc의 발현을 감소시킬 수 있으며, 사이클린-의존성 키나아제(cdk) 억제제 p27kipl의 발현을 감소시켜서, cdk2 활성을 억제시킴으로써 세포증식을 유도한다. The anti-Her2 antibody may be, for example, pertuzumab. Pertuzumab herein is a monoclonal antibody that targets HER2 and is marketed as Perjeta®. Pertuzumab binds to region II of the extracellular matrix domain of the HER2 receptor and inhibits the formation of dimers, thereby inhibiting the receptor-mediated signaling pathway. ErbB2, also known as HER2/neu, is the second member of the EGFR family and exhibits biological functions by forming a heterodimer with the other three members of the EGFR family, EGFR/HER1, HER3 and HER4. The neu gene encoding ErbB2 was first isolated from rat neuroblastoma. The homologous gene of the neu gene in human somatic cells, also known as HER2, is located on chromosome 17q21.1. The encoded product is ErbB2, consisting of 1255 amino acids having a molecular weight of approximately 185 kDa, and the 720 to 987 amino acid positions belong to the tyrosine kinase active domain. ErbB2 is dependent on the PI3K and MAPK signaling pathways, can reduce the expression of cyclin D and c-myc, and decreases the expression of the cyclin-dependent kinase (cdk) inhibitor p27kipl, inducing cell proliferation by inhibiting cdk2 activity. do.
"항체"는 생물학적 활성을 중화, 차단, 억제, 제거, 감소 또는 방해할 수 있는 단백질을 의미한다. "단클론성 항체"는 실질적으로 동일한 항체 집단에서 수득한 단일 분자 조성의 항체 분자, 즉 집단을 차지하고 있는 개개의 항체가 미량으로 존재할 수 있는 가능한 천연 발생적 돌연변이를 제외하고는 동일한 것을 의미한다. 단일클론항체는 고도로 특이적이어서, 단일 항원 부위에 대항하여 유도되며, 특정 에피토프에 대해 단일 결합 특이성 및 친화도를 나타낸다. “Antibody” means a protein capable of neutralizing, blocking, inhibiting, eliminating, reducing or interfering with a biological activity. By “monoclonal antibody” is meant an antibody molecule of single molecular composition obtained from a population of substantially the same antibody, ie the individual antibodies occupying the population are identical except for possible naturally occurring mutations that may be present in trace amounts. Monoclonal antibodies are highly specific, so they are directed against a single antigenic site and exhibit a single binding specificity and affinity for a particular epitope.
상기 항체에는 완전한 항체 형태뿐 아니라, 항체의 항원 결합 단편도 포함된다. 완전한 항체는 2개의 전체 길이의 경쇄 및 2개의 전체 길이의 중쇄를 가지는 약 150kDa의 4량체 단백질이며, 각각의 경쇄는 중쇄와 다이설파이드 결합으로 연결되어 있다. The antibody includes not only the complete antibody form, but also the antigen-binding fragment of the antibody. A complete antibody is a tetrameric protein of about 150 kDa with two full-length light chains and two full-length heavy chains, each of which is linked to a heavy chain by a disulfide bond.
항체의 항원 결합 단편 또는 항체 단편은 Fab, F(ab'), F(ab')2, Fv 또는 이의에피토프-결합 단편 등을 포함할 수 있다. Antigen-binding fragments or antibody fragments of an antibody may include Fab, F(ab'), F(ab')2, Fv, or an epitope-binding fragment thereof.
"Fab"는 경쇄 및 중쇄의 가변영역과 경쇄의 불변영역 및 중쇄의 첫 번째 불변영역(CH1)을 가지는 구조로 1개의 항원 결합 부위를 가진다. "F(ab')"는 중쇄 CH1 도메인의 C-말단에 하나 이상의 시스테인 잔기를 포함하는 힌지 영역(hinge region)을 가진다는 점에서 Fab와 차이가 있다. "F(ab')2" 항체는 F(ab')의 힌지 영역의 시스테인 잔기가 디설파이드 결합을 이루면서 생성된다."Fab" is a structure having a variable region of a light and heavy chain, a constant region of a light chain, and a first constant region (CH1) of a heavy chain, and has one antigen-binding site. "F(ab')" differs from Fab in that it has a hinge region containing one or more cysteine residues at the C-terminus of the heavy chain CH1 domain. "F(ab')2" antibodies are produced by disulfide bonds between cysteine residues in the hinge region of F(ab').
"Fv" 단편은 완전한 항체 인식 및 결합 부위를 함유하는 항체 단편이다. 이중쇄 Fv(two-chain Fv)는 비공유 결합으로 중쇄 가변영역과 경쇄 가변영역이 연결되어 있고 단쇄 Fv(single-chain Fv, scFv)는 일반적으로 펩타이드 링커를 통하여 중쇄의 가변영역과 경쇄의 가변영역이 공유결합으로 연결되어 있다. “Fv” fragments are antibody fragments that contain a complete antibody recognition and binding site. The double-chain Fv (two-chain Fv) is a non-covalent bond where the heavy chain variable region and the light chain variable region are connected, and the single-chain Fv (scFv) is generally a heavy chain variable region and a light chain variable region through a peptide linker. It is connected by this covalent bond.
"중쇄"는 IgG 항체의 중쇄를 의미할 수 있고, 항원에 특이성을 부여하기 위한 가변영역 도메인 VH 및 3 개의 불변영역 도메인 CH1, CH2 및 CH3을 포함하는 전체길이 중쇄 및 이의 단편을 모두 의미한다. 또한, 본 명세서에서 사용되는 용어, "경쇄"는 IgG 항체의 경쇄를 의미할 수 있고, 항원에 특이성을 부여하기 위한 가변영역 도메인 VL 및 불변영역 도메인 CL을 포함하는 전체길이 경쇄 및 이의 단편을 모두 의미한다."Heavy chain" may refer to the heavy chain of an IgG antibody, and refers to both a full-length heavy chain including a variable region domain VH and three constant region domains CH1, CH2 and CH3 for conferring specificity to an antigen, and fragments thereof. In addition, the term "light chain" as used herein may mean the light chain of an IgG antibody, and all of the full-length light chain including the variable region domain VL and the constant region domain CL for conferring specificity to the antigen and fragments thereof it means.
"가변 도메인"은 상보성 결정 영역 (CDR; 즉, CDR1, CDR2, 및 CDR3), 및 골격 영역 (FR)의 아미노산 서열을 포함하는 항체 분자의 경쇄 및 중쇄 부분을 의미한다. VH는 중쇄의 가변 도메인을 지칭한다. VL은 경쇄의 가변 도메인을 의미한다.“Variable domain” refers to the light and heavy chain portions of an antibody molecule comprising the amino acid sequences of the complementarity determining regions (CDRs; ie, CDR1, CDR2, and CDR3), and framework regions (FR). VH refers to the variable domain of the heavy chain. VL refers to the variable domain of the light chain.
"상보성 결정 영역" (CDR; 즉, CDR1, CDR2, 및 CDR3)은 항원 결합을 위해 필요한, 항체 가변 도메인의 아미노산 잔기를 의미한다. 중쇄 및 경쇄 가변 도메인 각각은 전형적으로, CDR1, CDR2 및 CDR3의 3개의 CDR 영역을 가진다.“Complementarity determining region” (CDR; ie, CDR1, CDR2, and CDR3) refers to the amino acid residues of an antibody variable domain, which are required for antigen binding. Each of the heavy and light chain variable domains typically has three CDR regions: CDR1, CDR2 and CDR3.
"골격 영역" (FR)은 CDR 잔기 이외의 가변 도메인 잔기이다. 각 가변 도메인은 전형적으로, FR1, FR2, FR3 및 FR4로서 확인된 4개의 FR을 가진다.The “skeletal region” (FR) is a variable domain residue other than a CDR residue. Each variable domain typically has 4 FRs identified as FR1, FR2, FR3 and FR4.
중쇄 불변영역은 감마(γ), 뮤(μ), 알파(α), 델타(δ) 및 엡실론(ε) 타입을 가지고 서브클래스로 감마1(γ1), 감마2(γ2), 감마3(γ3), 감마4(γ4), 알파1(α1) 및 알파2(α2)를 가진다. 경쇄의 불변영역은 카파(κ) 및 람다(λ) 타입을 가진다.The heavy chain constant region has gamma (γ), mu (μ), alpha (α), delta (δ) and epsilon (ε) types, and has subclasses of gamma 1 (γ1), gamma 2 (γ2), and gamma 3 (γ3). ), gamma4(γ4), alpha1(α1) and alpha2(α2). The constant region of the light chain has kappa (κ) and lambda (λ) types.
"에피토프"는 항체가 특이적으로 결합할 수 있는 결정부위 (determinant)를 의미한다. 에피토프는 통상 화학적으로 활성인 표면 분자군, 예를 들어 아미노산 또는 당 측쇄로 구성되며, 일반적으로 특정한 3차원의 구조적 특징뿐만 아니라 특정한 전하 특성을 갖는다. "Epitope" refers to a determinant to which an antibody can specifically bind. Epitopes are usually composed of a group of chemically active surface molecules, such as amino acids or sugar side chains, and generally have specific three-dimensional structural characteristics as well as specific charge characteristics.
단일클론항체는 실질적으로 동질적 항체 집단으로부터 수득한 항체, 즉 집단을 차지하고 있는 개개의 항체가 미량으로 존재할 수 있는 가능한 천연 발생적 돌연변이를 제외하고는 동일한 것을 지칭한다. 단일클론 항체는 고도로 특이적이어서, 단일 항원 부위에 대항하여 유도된다.Monoclonal antibodies refer to antibodies obtained from a population of substantially homogeneous antibodies, i.e., those that are identical except for possible naturally occurring mutations in which the individual antibodies occupying the population may be present in trace amounts. Monoclonal antibodies are highly specific and are thus directed against a single antigenic site.
"인간화" 형태의 비-인간 (예: 뮤린) 항체는 비-인간 면역글로불린으로부터 유래된 최소 서열을 함유하는 키메라 항체이다. 인간화 항체는 수용자의 초가변 영역 잔기를 목적하는 특이성, 친화성 및 능력을 보유하고 있는 비-인간 종 (공여자 항체), 예를 들어 마우스, 랫트, 토끼 또는 비-인간 영장류의 초가변 영역 잔기로 대체시킨 인간 면역글로불린 (수용자 항체)이다. A non-human (eg murine) antibody in its “humanized” form is a chimeric antibody that contains a minimal sequence derived from a non-human immunoglobulin. Humanized antibodies are those of a non-human species (donor antibody), such as a mouse, rat, rabbit or non-human primate, that retain the desired specificity, affinity and ability of the hypervariable region residues of the recipient. It is a replaced human immunoglobulin (receptor antibody).
본 발명에 따른 퍼투주맙의 중쇄 CDR, 경쇄 CDR, VH 및 VL 서열은 US7862817B2 또는 US7537931B2에 기재되어 있으며, 해당 문헌은 참조로서 도입될 수 있다. 이외에도, 퍼투주맙의 산성 변이체 또는 이황화 환원 변이체를 포함하는 퍼투주맙의 비결합 시스테인 변이체 (unpaired cysteine variants) 또는 퍼투주맙의 VHS 연장 변이체가 포함될 수 있으며, 퍼투주맙의 산성 변이체에 대하여는 US8652474B2 또는 US9181346B2, 산성 변이체 또는 이황화 환원 변이체를 포함하는 퍼투주맙의 비결합 시스테인 변이체 (unpaired cysteine variants)에 대하여는 US9815904B2, US9969811B, 퍼투주맙의 VHS 연장 변이체 (VHS extension variants)에 대하여는 US7560111B2, US8241630B2 또는 US7879325B2에 각각 기재되어 있고, 해당 문헌 역시 참조로서 도입될 수 있다.The heavy chain CDR, light chain CDR, VH and VL sequences of Pertuzumab according to the present invention are described in US7862817B2 or US7537931B2, which documents can be incorporated by reference. In addition, unpaired cysteine variants of pertuzumab or VHS extension variants of pertuzumab including acidic variants or disulfide-reducing variants of pertuzumab may be included, and US8652474B2 or US9181346B2, acidic The unpaired cysteine variants of Pertuzumab, including variants or disulfide reducing variants, are described in US9815904B2, US9969811B, and VHS extension variants of Pertuzumab in US7560111B2, US8241630B2 or US7879325B2, respectively, This document may also be incorporated by reference.
항-Her2 항체 예를 들어, 퍼투주맙 항체의 제조는 통상의 항체 제조방법 예를 들어, 퍼투주맙 항체 코딩 핵산을 복제 가능한 벡터 내로 삽입하고, 벡터를 세포에 형질감염한 다음, 세포를 배양하고 배양된 세포에서 항체를 회수하는 방법을 통해 제조될 수 있다. 퍼투주맙 항체를 제조하는 방법은 US 7879325 또는 US 8241630에 구체적으로 기재되어 있으며, 해당 문헌은 참조로서 도입될 수 있다. Preparation of an anti-Her2 antibody, e.g., Pertuzumab antibody, involves a conventional antibody preparation method, e.g., inserting a nucleic acid encoding a pertuzumab antibody into a replicable vector, transfecting the vector into cells, and then culturing the cells and culturing them. It can be prepared through a method of recovering the antibody from the cell. A method for preparing a Pertuzumab antibody is specifically described in US 7879325 or US 8241630, and the document can be incorporated by reference.
상기 서열과 85% 이상, 구체적으로는 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% 이상, 또는 100% 동일한 서열을 포함하는 퍼투주맙 항체가 포함될 수 있다. 서열의 동일성은 최적으로 정렬된 두 서열을 비교하여 결정할 수 있고, 두 서열의 최적의 정렬을 위하여 기준 서열 (부가 또는 결실을 포함하지 않음)과 비교하여 부가 또는 결실(즉, 갭(gap))을 포함할 수 있다. 동일성은 백분율로 표시되며, 백분율은 동일한 핵산 염기 또는 아미노산 잔기가 둘 모두의 서열에서 나타나는 위치의 수를 결정하여 매칭된 위치의 수를 생성하고, 매칭된 위치의 수를 총 위치수로 나누고, 그 결과에 100을 곱하여 계산될 수 있다. 서열 동일성 백분율은 예를 들어 니들(Needle) 프로그램을 사용하여, BLOSUM62 매트릭스(matrix) 프로그램 등을 사용하여 결정될 수 있다. Perturb comprising a sequence that is 85% or more, specifically 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more, or 100% identical to the sequence Zumab antibodies may be included. The identity of the sequences can be determined by comparing two sequences that are optimally aligned, and additions or deletions (i.e., gaps) compared to a reference sequence (not including additions or deletions) for optimal alignment of the two sequences. It may include. Identity is expressed as a percentage, and the percentage determines the number of positions in which the same nucleic acid base or amino acid residue appears in both sequences to generate the number of matched positions, and the number of matched positions is divided by the total number of positions, and the It can be calculated by multiplying the result by 100. The percent sequence identity can be determined, for example, using the Needle program, the BLOSUM62 matrix program, and the like.
본 발명에서 항-Her2 항체는 질환의 효과적인 치료 또는 예방에 유효한 양으로 포함될 수 있다. 이에 따라, 항-Her2 항체는 20 mg/ml 내지 150mg/ml의 농도로 포함될 수 있다. 본 발명에 따른 제형에서 항-Her2 항체는 25 mg/ml 내지 130mg/ml, 30 mg/ml 내지 110mg/ml, 또는 40 mg/ml 내지 100mg/ml일 수 있다. 구체적으로, 항-Her2 항체는 약 20 mg/ml, 약 21 mg/ml, 약 22 mg/ml, 약 23 mg/ml, 약 24 mg/ml, 약 25 mg/ml, 약 26 mg/ml, 약 27 mg/ml, 약 28 mg/ml, 약 29 mg/ml, 약 30 mg/ml, 약 31 mg/ml, 약 32 mg/ml, 약 33 mg/ml, 약 34 mg/ml, 약 35 mg/ml, 약 36 mg/ml, 약 37 mg/ml, 약 38 mg/ml, 약 39 mg/ml, 약 40 mg/ml, 약 41 mg/ml, 약 42 mg/ml, 약 43 mg/ml, 약 44 mg/ml, 약 45 mg/ml, 약 46 mg/ml, 약 47 mg/ml, 약 48 mg/ml, 약 49 mg/ml, 약 50 mg/ml, 약 51 mg/ml, 약 52 mg/ml, 약 53 mg/ml, 약 54 mg/ml, 약 55 mg/ml, 약 55 mg/ml, 약 56 mg/ml, 약 57 mg/ml, 약 58 mg/ml, 약 59 mg/ml, 약 60 mg/ml, 약 61 mg/ml, 약 62 mg/ml, 약 63 mg/ml, 약 64 mg/ml, 약 65 mg/ml, 약 66 mg/ml, 약 67 mg/ml, 약 68 mg/ml, 약 69 mg/ml, 약 70 mg/ml, 약 71 mg/ml, 약 72 mg/ml, 약 73 mg/ml, 약 74 mg/ml, 약 75 mg/ml, 약 76 mg/ml, 약 77 mg/ml, 약 78 mg/ml, 약 79 mg/ml, 약 80 mg/ml, 약 81 mg/ml, 약 82 mg/ml, 약 83 mg/ml, 약 84 mg/ml, 약 85 mg/ml, 약 86 mg/ml, 약 87 mg/ml, 약 88 mg/ml, 약 89 mg/ml, 약 90 mg/ml, 약 91 mg/ml, 약 92 mg/ml, 약 93 mg/ml, 약 94 mg/ml, 약 95 mg/ml, 약 96 mg/ml, 약 97 mg/ml, 약 98 mg/ml, 약 99 mg/ml, 약 100 mg/ml, 약 101 mg/ml, 약 102 mg/ml, 약 103 mg/ml, 약 104 mg/ml, 약 105 mg/ml, 약 106 mg/ml, 약 107 mg/ml, 약 108 mg/ml, 약 109 mg/ml, 약 110 mg/ml, 약 111 mg/ml, 약 112 mg/ml, 약 113 mg/ml, 약 114 mg/ml, 약 115 mg/ml, 약 116 mg/ml, 약 117 mg/ml, 약 118 mg/ml, 약 119 mg/ml, 약 120 mg/ml, 약 121 mg/ml, 약 122 mg/ml, 약 123 mg/ml, 약 124 mg/ml, 약 125 mg/ml, 약 126 mg/ml, 약 127 mg/ml, 약 128 mg/ml, 약 129 mg/ml, 약 130 mg/ml, 약 131 mg/ml, 약 132 mg/ml, 약 133 mg/ml, 약 134 mg/ml, 약 135 mg/ml, 약 136 mg/ml, 약 137 mg/ml, 약 138 mg/ml, 약 139 mg/ml, 약 140 mg/ml, 약 141 mg/ml, 약 142 mg/ml, 약 143 mg/ml, 약 144 mg/ml, 약 145 mg/ml, 약 146 mg/ml, 약 147 mg/ml, 약 148 mg/ml, 약 149 mg/ml, 또는 약 150 mg/ml (기재된 수치 사이의 값도 포함될 수 있음)의 농도로 포함될 수 있다. 구체적 실시예에서, 본 발명은 20 mg/ml 내지 30mg/ml의 농도로 항-Her2 항체를 포함하는 IV 투여용 제형일 수 있다. 본 명세서에서 정량적 수치 전단에 기재된 "약"이 사용되는 경우, 수치 자체뿐 아니라, 수치를 기준으로 ± 10% 범위 내, ± 5% 범위 내 또는 ± 1% 범위 내의 수치 역시 포함하는 것으로 해석될 수 있다. 다르게 정의되지 않은 이상, "약"이 사용되는 경우 기재된 수치에 대하여 ± 10% 범위 내, ± 5% 범위 내 또는 ± 1% 범위 내의 변동을 포함하는 것으로 할 수 있다. In the present invention, the anti-Her2 antibody may be included in an amount effective for effective treatment or prevention of a disease. Accordingly, the anti-Her2 antibody may be included in a concentration of 20 mg/ml to 150 mg/ml. In the formulation according to the present invention, the anti-Her2 antibody may be 25 mg/ml to 130 mg/ml, 30 mg/ml to 110 mg/ml, or 40 mg/ml to 100 mg/ml. Specifically, the anti-Her2 antibody is about 20 mg/ml, about 21 mg/ml, about 22 mg/ml, about 23 mg/ml, about 24 mg/ml, about 25 mg/ml, about 26 mg/ml, About 27 mg/ml, about 28 mg/ml, about 29 mg/ml, about 30 mg/ml, about 31 mg/ml, about 32 mg/ml, about 33 mg/ml, about 34 mg/ml, about 35 mg/ml, about 36 mg/ml, about 37 mg/ml, about 38 mg/ml, about 39 mg/ml, about 40 mg/ml, about 41 mg/ml, about 42 mg/ml, about 43 mg/ ml, about 44 mg/ml, about 45 mg/ml, about 46 mg/ml, about 47 mg/ml, about 48 mg/ml, about 49 mg/ml, about 50 mg/ml, about 51 mg/ml, About 52 mg/ml, about 53 mg/ml, about 54 mg/ml, about 55 mg/ml, about 55 mg/ml, about 56 mg/ml, about 57 mg/ml, about 58 mg/ml, about 59 mg/ml, about 60 mg/ml, about 61 mg/ml, about 62 mg/ml, about 63 mg/ml, about 64 mg/ml, about 65 mg/ml, about 66 mg/ml, about 67 mg/ ml, about 68 mg/ml, about 69 mg/ml, about 70 mg/ml, about 71 mg/ml, about 72 mg/ml, about 73 mg/ml, about 74 mg/ml, about 75 mg/ml, About 76 mg/ml, about 77 mg/ml, about 78 mg/ml, about 79 mg/ml, about 80 mg/ml, about 81 mg/ml, about 82 mg/ml, about 83 mg/ml, about 84 mg/ml, about 85 mg/ml, about 86 mg/ml, about 87 mg/ml, about 88 mg/ml, about 89 mg/ml, about 90 mg/ml, about 91 mg/ml, about 92 mg/ ml, about 93 mg/ml, about 94 mg/ml, about 95 mg/ml, about 96 mg/ml, about 97 mg/ml, about 98 mg/ml, about 99 mg/ml, about 100 mg/m l, about 101 mg/ml, about 102 mg/ml, about 103 mg/ml, about 104 mg/ml, about 105 mg/ml, about 106 mg/ml, about 107 mg/ml, about 108 mg/ml, About 109 mg/ml, about 110 mg/ml, about 111 mg/ml, about 112 mg/ml, about 113 mg/ml, about 114 mg/ml, about 115 mg/ml, about 116 mg/ml, about 117 mg/ml, about 118 mg/ml, about 119 mg/ml, about 120 mg/ml, about 121 mg/ml, about 122 mg/ml, about 123 mg/ml, about 124 mg/ml, about 125 mg/ ml, about 126 mg/ml, about 127 mg/ml, about 128 mg/ml, about 129 mg/ml, about 130 mg/ml, about 131 mg/ml, about 132 mg/ml, about 133 mg/ml, About 134 mg/ml, about 135 mg/ml, about 136 mg/ml, about 137 mg/ml, about 138 mg/ml, about 139 mg/ml, about 140 mg/ml, about 141 mg/ml, about 142 mg/ml, about 143 mg/ml, about 144 mg/ml, about 145 mg/ml, about 146 mg/ml, about 147 mg/ml, about 148 mg/ml, about 149 mg/ml, or about 150 mg May be included as a concentration of /ml (values between listed values may also be included). In a specific embodiment, the present invention may be a formulation for IV administration containing an anti-Her2 antibody at a concentration of 20 mg/ml to 30 mg/ml. When the "about" described in the front end of the quantitative numerical value in the present specification is used, it may be interpreted as including not only the value itself, but also within the range of ± 10%, within the range of ± 5%, or within the range of ± 1% based on the numerical value. have. Unless otherwise defined, when "about" is used, it may be said to include fluctuations within the range of ± 10%, within the range of ± 5%, or within the range of ± 1% with respect to the stated value.
본 발명은 또 다른 관점에서, 항-Her2 항체 및 2종 이상의 아미노산을 포함하고, 아세트산염을 포함하지 않으며, 고온 보관 조건, 냉동/해동 조건 또는 교반 조건에서 안정성이 유지 또는 향상된 제형에 관한 것이다. In another aspect, the present invention relates to a formulation containing an anti-Her2 antibody and two or more amino acids, no acetate, and maintaining or improving stability under high temperature storage conditions, freezing/thawing conditions, or stirring conditions.
경우에 따라서, 본 발명에 따른 제형은 당 또는 당 알코올을 추가로 포함할 수 있다.In some cases, the formulation according to the present invention may further contain sugar or sugar alcohol.
상기 당 또는 당 알코올은 예를 들어, 글루코스, 수크로오스, 트레할로스, 락토오스, 프럭토오스, 말토오스, 덱스트란, 글리세린, 에리트리톨, 아라비톨, 자일리톨, 솔비톨, 만니톨, 멜리바이오스, 멜레치토스, 라피노오스, 만노트리오스, 스타키오스, 말토오스, 락툴로오스, 말툴로오스, 글루시톨, 말티톨, 락티톨, 이소-말툴로오스로 구성된 군에서 선택될 수 있다. 본 발명에 따른 구체예에서, 수크로오스, 솔비톨 또는 트레할로스를 포함하는 제형에서 항체의 고온 보관 조건, 냉동-해동 조건 또는 교반 조건 중 항-Her2 항체가 안정성을 나타냄을 확인하였다. The sugar or sugar alcohol is, for example, glucose, sucrose, trehalose, lactose, fructose, maltose, dextran, glycerin, erythritol, arabitol, xylitol, sorbitol, mannitol, melibiose, meletithose, raffinose. , Mannotriose, stachiose, maltose, lactulose, maltulose, glucitol, maltitol, lactitol, iso-maltulose may be selected from the group consisting of. In the embodiment according to the present invention, it was confirmed that the anti-Her2 antibody exhibited stability under high temperature storage conditions, freeze-thaw conditions, or agitation conditions of the antibody in the formulation containing sucrose, sorbitol or trehalose.
"안정"한 제형은 기계적 스트레스, 열적 스트레스 또는 냉동 및 해동에서 생기는 스트레스로부터 항-Her2 항체의 물리적 안정성, 화학적 안정성 또는 생물학적 활성을 유지하는 것으로, 선택된 기간 동안 선택된 온도에서 예를 들어, 약 5±3℃, 약 25±2℃ 또는 약 40±2℃에서 적어도 약 1, 2, 3, 4, 5, 6일, 1주, 2주, 3주, 4주, 5주, 6주, 7주, 8주, 9주, 10주, 11주, 12주, 13주, 14주, 15주, 16주, 17주, 18주, 19주, 20주, 21주, 22주, 23주, 24주 또는 그 이상의 기간 동안 안정하거나, 동결 (예를 들어, -20℃ 또는 -70℃) 및 해동 후 예를 들어, 1, 2, 3, 4, 5, 또는 그 이상의 사이클로 동결 및 해동한 후에 안정하거나, 교반 스트레스 조건 예를 들어 100 rpm 이상, 200 rpm 이상, 300 rpm 이상, 400 rpm 이상, 500 rpm 이상의 조건에서 안정한 것을 의미한다. A “stable” formulation is one that maintains the physical stability, chemical stability or biological activity of the anti-Her2 antibody from mechanical stress, thermal stress, or stress resulting from freezing and thawing, at a selected temperature for a selected period of time, for example about 5± At least about 1, 2, 3, 4, 5, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks at 3℃, about 25±2℃ or about 40±2℃ , 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 Stable for a period of weeks or longer, or after freezing (eg -20°C or -70°C) and thawing, eg, after freezing and thawing in 1, 2, 3, 4, 5, or more cycles Or, it means that it is stable under agitation stress conditions, for example, 100 rpm or more, 200 rpm or more, 300 rpm or more, 400 rpm or more, and 500 rpm or more.
응집 형성의 평가 (예를 들어, 크기 배제 크로마토그래피를 이용하거나 탁도 측정 및/또는 육안 검사), 양이온 교환 크로마토그래피, 이미지 모세관 등전점 포커싱 (icIEF) 또는 모세관 구역 전기영동을 이용한 전하 이종성 평가, 아미노-말단 또는 카르복시-말단 서열 분석, 질량 분광측정 분석, 환원된 항체와 무손상 항체를 비교하는 SDS-PAGE 분석, 펩티드 맵 (예를 들어, 트립신 또는 LYS-C) 분석, 또는 항체의 생물학적 활성 또는 항원 결합 기능 평가 등을 포함하는 다양한 방법을 통해 안정성을 평가할 수 있다. Assessment of aggregation formation (e.g., using size exclusion chromatography or measuring turbidity and/or visual inspection), cation exchange chromatography, evaluation of charge heterogeneity using image capillary isoelectric focusing (icIEF) or capillary zone electrophoresis, amino- Terminal or carboxy-terminal sequence analysis, mass spectrometric analysis, SDS-PAGE analysis comparing reduced and intact antibodies, peptide map (e.g., trypsin or LYS-C) analysis, or biological activity or antigen of the antibody Stability can be evaluated through various methods including evaluation of binding function and the like.
색상 및/또는 투명도의 육안 검사, UV 광산란 또는 크기 배제 크로마토그래피 (SEC) 등에 의하여 항체가 응집, 침전, 및/또는 변성되지 않은 경우 물리적 안정성을 유지함을 의미할 수 있다. When the antibody is not aggregated, precipitated, and/or denatured by visual inspection of color and/or transparency, UV light scattering or size exclusion chromatography (SEC), it may mean maintaining physical stability.
절단 또는 결합 형성에 의해 항체가 변형되지 않는다면, 항체는 화학적 안정성을 유지함을 의미할 수 있다. 화학적 안정성은 항체가 화학적으로 변형된 형태를 검출 및 정량함으로써 확인될 수 있다. If the antibody is not modified by cleavage or bond formation, it may mean that the antibody remains chemically stable. Chemical stability can be ascertained by detecting and quantifying the chemically modified form of the antibody.
항원 결합 분석 또는 중화 분석에 의하여 항체의 생물학적 활성이 제형 제조시 항체 생물학적 활성의 약 70% 내지 약 150%, 약 80% 내지 약 125%, 또는 약 90% 내지 약 110%인 경우, 생물학적 활성을 유지함을 의미할 수 있다. 특정 조건 및 시간에서 항체의 생물학적 활성이 제형 제조시 항체 생물학적 활성의 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 100%, 105%, 110%, 115%, 120% 또는 125%인 경우 생물학적 활성을 유지할 수 있다. When the biological activity of the antibody by antigen binding assay or neutralization assay is about 70% to about 150%, about 80% to about 125%, or about 90% to about 110% of the antibody biological activity at the time of preparation of the formulation, the biological activity is It can mean keeping. The biological activity of the antibody under certain conditions and times is 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 100% of the antibody biological activity at the time of preparation of the formulation, If it is 105%, 110%, 115%, 120% or 125%, biological activity can be maintained.
항-Her2 항체의 생물학적 활성은 예를 들어, Her2 결합 수준을 통해 확인할 수 있다. 구체적으로, 항-Her2 항체의 생물학적 활성은 수용체 결합 활성 (Receptor-Binding Activity: RBA)을 통해 측정될 수 있다. The biological activity of an anti-Her2 antibody can be ascertained, for example, through the level of Her2 binding. Specifically, the biological activity of the anti-Her2 antibody can be measured through Receptor-Binding Activity (RBA).
하나의 구체예에서, 본 발명에 따른 제형 중 40℃±2℃ 8주 보관 조건에서 수용체 결합 활성을 확인하여 측정된 Her2결합 수준은 예를 들어 92% 이상, 구체적으로 92 내지 97%일 수 있다. 40℃±2℃ 12주 보관 조건에서 수용체 결합 활성을 확인하여 측정된 Her2 결합 수준은 예를 들어 90% 이상, 구체적으로 90 내지 95%일 수 있다. In one embodiment, the Her2 binding level measured by confirming the receptor binding activity under storage conditions of 40°C±2°C for 8 weeks in the formulation according to the present invention may be, for example, 92% or more, specifically 92 to 97%. . The Her2 binding level measured by confirming the receptor-binding activity under storage conditions of 40°C±2°C for 12 weeks may be, for example, 90% or more, specifically 90 to 95%.
다른 구체예에서, 본 발명에 따른 제형 중 5℃±3℃ 8주 보관 조건에서 수용체 결합 활성을 확인하여 측정된 Her2 결합 수준은 예를 들어 97% 이상, 구체적으로 97 내지 99%일 수 있다. In another embodiment, the Her2 binding level measured by confirming the receptor binding activity under storage conditions of 5°C±3°C for 8 weeks in the formulation according to the present invention may be, for example, 97% or more, specifically 97 to 99%.
또 다른 구체예에서, 본 발명에 따른 제형 중 -70±10℃ 1 사이클 이상, 예를 들어 5 사이클의 냉동/해동 주기의 제형에서 수용체 결합 활성을 확인하여 측정된 Her2결합 수준은 예를 들어 97% 이상, 구체적으로 97 내지 99%일 수 있다. In another embodiment, the Her2 binding level measured by confirming the receptor binding activity in the formulation of -70±10° C. 1 cycle or more, for example 5 cycles of freeze/thaw cycle in the formulation according to the present invention, is, for example, 97 % Or more, specifically 97 to 99%.
하나의 구현예에서, 제형 중 항체의 안정성은 제형 중 응집도 (aggregation) 또는 조각화 수준 (fragment levels)을 확인하여 결정될 수 있다. 제형 중 고분자량 생성물 (HMW: High molecular weight species) 및/또는 저분자량 생성물 (LMW: Low molecular weight species)이 형성되는지 여부를 측정하여 결정될 수 있다. 특정 온도 조건에서 제형 중 고분자량 생성물 및/또는 저분자량 생성물의 함량을 측정할 수 있다. 상기 고분자량 생성물 및/또는 저분자량 생성물의 함량은 크기 배제 HPLC(SE-HPLC)에 의해 측정될 수 있다. In one embodiment, the stability of the antibody in the formulation can be determined by identifying aggregation or fragment levels in the formulation. It can be determined by measuring whether a high molecular weight product (HMW) and/or a low molecular weight species (LMW) are formed in the formulation. The content of the high molecular weight product and/or the low molecular weight product in the formulation can be measured under certain temperature conditions. The content of the high molecular weight product and/or the low molecular weight product can be measured by size exclusion HPLC (SE-HPLC).
하나의 구현예에서, 제형 중 항체의 안정성은 제형 중 고분자량 생성물 (HMW)이 형성되는지 여부를 측정하여 결정될 수 있다. 특정 온도 조건에서 제형 중 HMW의 변화량을 측정할 수 있으며, 상기 HMW는 크기 배제 HPLC(SE-HPLC)에 의해 측정될 수 있다. In one embodiment, the stability of the antibody in the formulation can be determined by measuring whether a high molecular weight product (HMW) is formed in the formulation. The amount of change in HMW in the formulation can be measured under specific temperature conditions, and the HMW can be measured by size exclusion HPLC (SE-HPLC).
약 5±3℃, 25±2℃ 또는 40±2℃의 온도에서 1주 이상, 2주 이상, 3주 이상, 4주 이상, 5주 이상, 6주 이상, 7주 이상, 8주 이상의 기간 동안 제형 중 크기 배제 HPLC(SE-HPLC)를 통해 측정된 HMW의 함량%는 6.0% 이하, 5.0% 이하, 4.0% 이하, 3.0% 이하, 2.0% 이하, 1.0% 이하, 0.9% 이하, 0.8% 이하, 0.7% 이하, 0.6% 이하, 0.5% 이하, 0.4% 이하, 0.3% 이하, 0.2% 이하 또는 0.1% 이하일 수 있다. 1 week or more, 2 weeks or more, 3 weeks or more, 4 weeks or more, 5 weeks or more, 6 weeks or more, 7 weeks or more, 8 weeks or more at a temperature of about 5±3℃, 25±2℃ or 40±2℃ During the formulation, the% content of HMW measured by size exclusion HPLC (SE-HPLC) was 6.0% or less, 5.0% or less, 4.0% or less, 3.0% or less, 2.0% or less, 1.0% or less, 0.9% or less, 0.8% It may be 0.7% or less, 0.6% or less, 0.5% or less, 0.4% or less, 0.3% or less, 0.2% or less, or 0.1% or less.
하나의 구체예에서, 본 발명에 따른 제형 중 40℃±2℃ 1주 이상의 보관 조건에서 크기 배제 HPLC(SE-HPLC)를 통해 측정된 HMW는 0.5% 이하일 수 있다. In one embodiment, the HMW measured by size exclusion HPLC (SE-HPLC) at 40° C.±2° C. for 1 week or more in the formulation according to the present invention may be 0.5% or less.
구체적 실시예에서, 40℃±2℃의 온도 1주 보관 조건에서 제형 중 크기 배제 HPLC(SE-HPLC)를 통해 측정된 HMW의 함량%는 예를 들어 0.27% 이하, 구체적으로 0.16 내지 0.27%일 수 있다. 40℃±2℃의 온도 2주 보관 조건에서 제형 중 크기 배제 HPLC(SE-HPLC)를 통해 측정된 HMW의 함량%는 예를 들어 0.33% 이하, 구체적으로 0.17 내지 0.33%일 수 있다. 40℃±2℃의 온도 4주 보관 조건에서 제형 중 크기 배제 HPLC(SE-HPLC)를 통해 측정된 HMW의 함량%는 예를 들어 0.4% 이하, 구체적으로 0.20 내지 0.4%일 수 있다. 40℃±2℃의 온도 8주 보관 조건에서 제형 중 크기 배제 HPLC(SE-HPLC)를 통해 측정된 HMW의 함량%는 예를 들어 0.48% 이하, 구체적으로 0.24 내지 0.48%일 수 있다.In specific examples, the% of HMW content measured through size exclusion HPLC (SE-HPLC) in the formulation under storage conditions at a temperature of 40° C.±2° C. for 1 week is, for example, 0.27% or less, specifically 0.16 to 0.27%. I can. The content% of HMW measured through size exclusion HPLC (SE-HPLC) in the formulation under storage conditions at a temperature of 40°C±2°C for 2 weeks may be, for example, 0.33% or less, specifically 0.17 to 0.33%. The content% of HMW measured through size exclusion HPLC (SE-HPLC) in the formulation under storage conditions at a temperature of 40°C±2°C for 4 weeks may be, for example, 0.4% or less, specifically 0.20 to 0.4%. The content% of HMW measured through size exclusion HPLC (SE-HPLC) in the formulation under storage conditions at a temperature of 40°C±2°C for 8 weeks may be, for example, 0.48% or less, and specifically 0.24 to 0.48%.
다른 실시예에서, 40℃±2℃의 온도 1주 보관 조건에서 제형 중 크기 배제 HPLC(SE-HPLC)를 통해 측정된 HMW의 함량%는 예를 들어 0.2% 이하, 구체적으로 0.15 내지 0.2%일 수 있다. 40℃±2℃의 온도 2주 보관 조건에서 제형 중 크기 배제 HPLC(SE-HPLC)를 통해 측정된 HMW의 함량%는 예를 들어 0.25% 이하, 구체적으로 0.2 내지 0.25%일 수 있다. 40℃±2℃의 온도 8주 보관 조건에서 제형 중 크기 배제 HPLC(SE-HPLC)를 통해 측정된 HMW의 함량%는 예를 들어 0.5% 이하, 구체적으로 0.35 내지 0.5%일 수 있다. 40℃±2℃의 온도 12주 보관 조건에서 제형 중 크기 배제 HPLC(SE-HPLC)를 통해 측정된 HMW의 함량%는 예를 들어 0.6% 이하, 구체적으로 0.35 내지 0.6%일 수 있다.In another example, the HMW content% measured through size exclusion HPLC (SE-HPLC) in the formulation at a temperature of 40° C.±2° C. for 1 week storage conditions is, for example, 0.2% or less, specifically 0.15 to 0.2%. I can. The content% of HMW measured through size exclusion HPLC (SE-HPLC) in the formulation at a temperature of 40°C±2°C for 2 weeks storage may be, for example, 0.25% or less, specifically 0.2 to 0.25%. The content% of HMW measured through size exclusion HPLC (SE-HPLC) in the formulation under storage conditions at a temperature of 40°C±2°C for 8 weeks may be, for example, 0.5% or less, specifically 0.35 to 0.5%. The content% of HMW measured through size exclusion HPLC (SE-HPLC) in the formulation under storage conditions at a temperature of 40°C±2°C for 12 weeks may be, for example, 0.6% or less, and specifically 0.35 to 0.6%.
-70±10℃ 1 사이클 이상의 냉동/해동 주기의 제형 중 크기 배제 HPLC(SE-HPLC)를 통해 측정된 HMW의 함량%는 2.0% 이하, 1.0% 이하, 0.9% 이하, 0.8% 이하, 0.7% 이하, 0.6% 이하, 0.5% 이하, 0.4% 이하, 0.3% 이하, 0.25% 이하, 0.2% 이하, 0.15% 이하 또는 0.1% 이하이다. 400 rpm 교반 조건에서 크기 배제 HPLC(SE-HPLC)를 통해 측정된 HMW가 0.25% 이하, 0.2% 이하, 0.15% 이하 또는 0.1% 이하일 수 있다.-70±10°C The content% of HMW measured by size exclusion HPLC (SE-HPLC) in the formulation of 1 cycle or more freeze/thaw cycle is 2.0% or less, 1.0% or less, 0.9% or less, 0.8% or less, 0.7% Or less, 0.6% or less, 0.5% or less, 0.4% or less, 0.3% or less, 0.25% or less, 0.2% or less, 0.15% or less, or 0.1% or less. HMW measured through size exclusion HPLC (SE-HPLC) under 400 rpm stirring conditions may be 0.25% or less, 0.2% or less, 0.15% or less, or 0.1% or less.
하나의 구체예에서, -70℃±10℃ 1 사이클 이상, 예를 들어 5 사이클의 냉동/해동 주기의 제형 중 크기 배제 HPLC(SE-HPLC)를 통해 측정된 HMW의 함량%는 예를 들어 2.0% 이하, 1.0% 이하, 0.9% 이하, 0.8% 이하, 0.7% 이하, 0.6% 이하, 0.5% 이하, 0.4% 이하, 0.3% 이하, 0.25% 이하, 0.2% 이하, 구체적으로 0.1 내지 0.2%일 수 있다. In one embodiment, the% content of HMW measured via size exclusion HPLC (SE-HPLC) in the formulation of 1 cycle or more, eg 5 cycles of freeze/thaw cycles, is, for example, 2.0 % Or less, 1.0% or less, 0.9% or less, 0.8% or less, 0.7% or less, 0.6% or less, 0.5% or less, 0.4% or less, 0.3% or less, 0.25% or less, 0.2% or less, specifically 0.1 to 0.2% I can.
다른 구체예에서, 400 rpm 교반 조건에서 제형 중 크기 배제 HPLC(SE-HPLC)를 통해 측정된 HMW의 함량%는 예를 들어 2.0% 이하, 1.0% 이하, 0.9% 이하, 0.8% 이하, 0.7% 이하, 0.6% 이하, 0.5% 이하, 0.4% 이하, 0.3% 이하, 0.25% 이하, 구체적으로 0.15 내지 0.25%일 수 있다.In another embodiment, the content% of HMW measured by size exclusion HPLC (SE-HPLC) in the formulation at 400 rpm stirring conditions is, for example, 2.0% or less, 1.0% or less, 0.9% or less, 0.8% or less, 0.7% It may be less than, 0.6% or less, 0.5% or less, 0.4% or less, 0.3% or less, 0.25% or less, specifically 0.15 to 0.25%.
5±3℃, 25±2℃ 또는 40±2℃의 온도에서 1주 이상, 2주 이상, 3주 이상, 4주 이상, 5주 이상, 6주 이상, 7주 이상, 8주 이상의 기간 동안 제형 중 항체 단량체의 함량%는 96% 이상, 97% 이상, 98% 이상, 99% 이상, 100%일 수 있다. -70±10℃ 1 사이클 이상의 냉동/해동 주기의 제형 중 항체 단량체의 함량%는 96% 이상, 97% 이상, 98% 이상, 99% 이상, 100%일 수 있다.For a period of 1 week or more, 2 weeks or more, 3 weeks or more, 4 weeks or more, 5 weeks or more, 6 weeks or more, 7 weeks or more, 8 weeks or more at a temperature of 5±3℃, 25±2℃ or 40±2℃ The content% of the antibody monomer in the formulation may be 96% or more, 97% or more, 98% or more, 99% or more, or 100%. In the formulation of -70±10° C. 1 cycle or more, the content of the antibody monomer may be 96% or more, 97% or more, 98% or more, 99% or more, or 100%.
하나의 구체예에서, 40℃±2℃ 8주 조건에서 CE-SDS (Capillary electrophoresis with sodium dodecylsulfate)를 통해 측정한 제형 중 IgG의 함량%는 예를 들어 90% 이상, 구체적으로 90% 내지 93%일 수 있다. 40℃±2℃ 12주 조건에서 CE-SDS를 통해 측정한 제형 중 IgG의 함량%는 예를 들어 87% 이상, 구체적으로 87% 내지 89%일 수 있다.In one embodiment, the percentage of the IgG content in the formulation measured by CE-SDS (Capillary electrophoresis with sodium dodecylsulfate) at 40°C±2°C for 8 weeks is, for example, 90% or more, specifically 90% to 93%. Can be The amount of IgG in the formulation measured through CE-SDS at 40° C.±2° C. for 12 weeks may be, for example, 87% or more, and specifically 87% to 89%.
다른 구체예에서, 5℃±3℃ 온도 8주 조건에서 CE-SDS를 통해 측정한 제형 중 IgG의 함량%는 예를 들어 95% 이상, 구체적으로 95% 내지 97%일 수 있다. 5℃±3℃ 12주 조건에서 CE-SDS를 통해 측정한 제형 중 IgG의 함량%는 예를 들어 94% 이상, 구체적으로 94% 내지 95%일 수 있다. 또 다른 구체예에서, -70℃±10℃ 1 사이클 이상, 예를 들어 5 사이클의 냉동/해동 주기의 제형 중 CE-SDS를 통해 측정한 IgG의 함량%는 예를 들어 95% 이상, 구체적으로 95% 내지 97%일 수 있다.In another embodiment, the% of the IgG content in the formulation measured through CE-SDS at a temperature of 5°C±3°C for 8 weeks may be, for example, 95% or more, specifically 95% to 97%. The percentage of the IgG content in the formulation measured through CE-SDS under conditions of 5°C±3°C for 12 weeks may be, for example, 94% or more, and specifically 94% to 95%. In another embodiment, the percentage of IgG content measured through CE-SDS in the formulation of 1 cycle or more, eg, 5 cycles of freeze/thaw cycle, is, for example, 95% or more, specifically It may be 95% to 97%.
또한, 제형 중 항체의 안정성은 항체 중 변이체가 형성되는지 여부를 측정하여 결정될 수 있다. 상기 변이체는 예를 들어, 주요 항체와 다른 아미노산 서열을 가지는 항체를 의미할 수 있으며, 항체의 아미노산 서열 내 또는 인접한 특정 위치에 아미노산의 치환, 결실 및/또는 부가를 포함할 수 있다. In addition, the stability of the antibody in the formulation can be determined by measuring whether a variant in the antibody is formed. The variant may mean, for example, an antibody having an amino acid sequence different from that of the main antibody, and may include substitution, deletion, and/or addition of amino acids at a specific position within or adjacent to the amino acid sequence of the antibody.
상기 변이체는 항체의 변형에 의해 전하가 달라지는 것을 포함할 수 있으며, 예를 들어 전하 변이체일 수 있다. 전하 변이체는 변형되지 않은 항체에 비해 산성을 나타낼 수 있고, 낮은 pI값을 가질 수 있다. 산성 변이체는 주요 항체보다 음성인 전하를 가지거나 양성인 전하를 잃게 되었을 경우 생성될 수 있다. 또한, 전하 변이체는 변형되지 않은 항체에 비해 염기성을 나타낼 수 있고, 높은 pI값을 가질 수 있다. 염기성 변이체는 주요 항체보다 양성인 전하를 가지거나 음성인 전하를 잃게 되었을 경우 생성될 수 있다. 이러한 전하 변이체는 항체의 산화, 탈아미드화, 라이신 잔기 C-말단의 가공, N-말단에 파이로글루타메이트 형성, 비효소적 당화에 의해 발생할 수 있다. 상기 전하 변이체는 전하에 따라 단백질을 분리하는 이온 교환 크로마토그래피 예를 들어, 양이온 교환 크로마토그래피에 의해 분리되거나, 이미지 모세관 등전점 포커싱 (icIEF)에 의해 전하 이종성이 확인될 수 있다. The variant may include a change in charge due to modification of the antibody, and may be, for example, a charge variant. Charged variants can be acidic and have low pI values compared to unmodified antibodies. Acidic variants can be produced when they have a negative charge or lose a positive charge than the main antibody. In addition, the charge variant may exhibit basicity compared to an unmodified antibody and may have a high pI value. Basic variants can be produced if they have a positive charge or lose a negative charge than the main antibody. These charge variants can occur by oxidation of antibodies, deamidation, processing of the C-terminus of the lysine residue, formation of pyroglutamate at the N-terminus, and non-enzymatic glycosylation. The charge variants may be separated by ion exchange chromatography, for example, cation exchange chromatography to separate proteins according to charge, or charge heterogeneity may be confirmed by image capillary isoelectric focusing (icIEF).
5±3℃, 25±2℃ 또는 40±2℃ 8주 보관 조건에서 1주 이상, 2주 이상, 3주 이상, 4주 이상, 5주 이상, 6주 이상, 7주 이상, 8주 이상의 기간 동안 양이온 교환 HPLC를 통해 측정된 산성 변이체 또는 염기성 변이체의 함량%는 50% 이하, 45% 이하, 40% 이하, 35% 이하, 30% 이하, 25% 이하, 20% 이하, 19% 이하, 18% 이하, 17% 이하, 15% 이하, 또는 10% 이하일 수 있다. 1 week or more, 2 weeks or more, 3 weeks or more, 4 weeks or more, 5 weeks or more, 6 weeks or more, 7 weeks or more, 8 weeks or more under storage conditions of 5±3℃, 25±2℃ or 40±2℃ for 8 weeks The content% of acidic variant or basic variant measured by cation exchange HPLC during the period is 50% or less, 45% or less, 40% or less, 35% or less, 30% or less, 25% or less, 20% or less, 19% or less, It may be 18% or less, 17% or less, 15% or less, or 10% or less.
선택적으로, 5±3℃, 25±2℃ 또는 40±2℃ 8주 보관 조건에서 1주 이상, 2주 이상, 3주 이상, 4주 이상, 5주 이상, 6주 이상, 7주 이상, 8주 이상의 기간 동안 이미지 모세관 등전점 포커싱 (icIEF: Imaged Capillary Isoelectric Focusing)에 의해 측정된 산성 변이체 또는 염기성 변이체의 함량%는 70% 이하, 65% 이하, 60% 이하, 55% 이하, 50% 이하, 또는 45% 이하일 수 있다. Optionally, 1 week or more, 2 weeks or more, 3 weeks or more, 4 weeks or more, 5 weeks or more, 6 weeks or more, 7 weeks or more under storage conditions of 5±3℃, 25±2℃ or 40±2℃ for 8 weeks, The content% of acidic variant or basic variant measured by Imaged Capillary Isoelectric Focusing (icIEF: Imaged Capillary Isoelectric Focusing) for a period of 8 weeks or longer is 70% or less, 65% or less, 60% or less, 55% or less, 50% or less, Or 45% or less.
구체적으로, icIEF를 사용하여 산성 변이체 비율을 분석한 경우, 산성변이체의 함량%는 1주 이상의 조건에서 70% 이하일 수 있다. Specifically, when an acidic variant ratio is analyzed using icIEF, the content% of the acidic variant may be 70% or less under conditions of 1 week or more.
하나의 구체예에서, 40℃±2℃의 온도 2주 보관 조건에서 icIEF에 의해 측정된 제형 중 산성변이체의 함량%는 예를 들어 45% 이하, 구체적으로 44.0 내지 44.5%일 수 있다. 40℃±2℃의 온도 4주 보관 조건에서 icIEF에 의해 측정된 제형 중 산성변이체의 함량%는 예를 들어 53% 이하, 구체적으로 50 내지 53%일 수 있다. 40℃±2℃의 온도 8주 보관 조건에서 icIEF에 의해 측정된 제형 중 산성변이체의 함량%는 예를 들어 65% 이하, 구체적으로 60 내지 65%일 수 있다. 40℃±2℃의 온도 12주 보관 조건에서 icIEF에 의해 측정된 제형 중 산성변이체의 함량%는 예를 들어 70% 이하, 구체적으로 65 내지 70%일 수 있다.In one embodiment, the content% of the acidic variant in the formulation measured by icIEF at a temperature of 40° C.±2° C. for 2 weeks storage may be, for example, 45% or less, and specifically 44.0 to 44.5%. The content% of the acidic variant in the formulation measured by icIEF at a temperature of 40°C±2°C for 4 weeks storage may be, for example, 53% or less, specifically 50 to 53%. The content% of the acidic variant in the formulation measured by icIEF at a temperature of 40°C±2°C for 8 weeks storage may be, for example, 65% or less, specifically 60 to 65%. The content% of the acidic variant in the formulation measured by icIEF at a temperature of 40°C±2°C for 12 weeks storage may be, for example, 70% or less, specifically 65 to 70%.
-70±10℃ 1 사이클 이상의 냉동/해동 주기의 제형 중 산성 변이체 또는 염기성 변이체의 함량%는 20% 이하, 18% 이하, 17% 이하, 15% 이하, 10% 이하, 5% 이하, 4% 이하, 3% 이하, 2% 이하, 1% 이하이다. 400 rpm 교반 조건에서 제형 중 산성 변이체 또는 염기성 변이체의 함량%는 20% 이하, 18% 이하, 17% 이하, 15% 이하, 10% 이하, 5% 이하, 4% 이하, 3% 이하, 2% 이하, 1% 이하이다. -70±10℃ The content% of acidic variant or basic variant in the formulation of 1 cycle or more of freeze/thaw cycle is 20% or less, 18% or less, 17% or less, 15% or less, 10% or less, 5% or less, 4% Or less, 3% or less, 2% or less, and 1% or less. The content% of acidic variant or basic variant in the formulation under 400 rpm stirring conditions is 20% or less, 18% or less, 17% or less, 15% or less, 10% or less, 5% or less, 4% or less, 3% or less, 2% It is less than or equal to 1%.
하나의 구체예에서, icIEF를 사용하여 산성 변이체 비율을 분석한 경우, -70℃±10℃ 냉동/해동 5사이클 조건에서 제형 중 산성변이체의 함량%는 예를 들어 38% 이하, 구체적으로 36 내지 38%일 수 있다. In one embodiment, when the acidic variant ratio is analyzed using icIEF, the content% of the acidic variant in the formulation under the conditions of -70 °C ± 10 °C freeze/thaw 5 cycles is, for example, 38% or less, specifically 36 to It could be 38%.
이에 따라, 본 발명에 따른 제형은 5℃±3℃ 이상, 특히 40℃±2℃에서 1개월 이상, 2개월 이상, 3개월 이상 또는 6개월 이상 동안 안정한 것으로 평가될 수 있다. 또한, 본 발명에 따른 제형은 -70℃±10℃ 냉동 및 해동 후 안정한 것으로 평가될 수 있으며, 냉동 및 해동은 -70±10℃에서 24시간 동안 냉동시킨 다음, 실온에서 90분 동안 해동시키는 과정을 포함하고, 상기 사이클은 1-5회 반복, 제5사이클에서 -70±10℃ 온도로 72시간 동안 유지될 수 있다. 더욱이, 400 rpm의 교반 조건에서 교반 후 안정한 것으로 평가될 수 있다.Accordingly, the formulation according to the present invention can be evaluated as stable for at least 1 month, at least 2 months, at least 3 months, or at least 6 months at 5°C±3°C or more, particularly 40°C±2°C. In addition, the formulation according to the present invention can be evaluated as stable after freezing and thawing at -70°C±10°C, freezing and thawing at -70±10°C for 24 hours, and then thawing at room temperature for 90 minutes Including, the cycle may be repeated 1-5 times, and maintained for 72 hours at a temperature of -70±10°C in the fifth cycle. Moreover, it can be evaluated as stable after stirring under the stirring conditions of 400 rpm.
본 발명에 따른 제형에서 항-Her2 항체는 치료적 유효량으로 포함될 수 있다. "유효량"은 HER2 수용체의 세포외기질 도메인의 영역 II에 결합하여 이량체의 형성을 억제하고, 이를 통해 수용체-매개성 신호경로를 저해하는데 필요하거나 충분한 양을 의미한다. 예를 들어, HER2의 이량체 형성과 관련된 상태의 다양한 형태학적 증상을 예방하거나 치료하는데 필요한 양이다. 증상이 암인 경우 암세포 수 감소, 종양 크기 감소, 암세포의 침윤 억제, 종양 전이 억제, 종양 성장 억제 또는 암의 증상 경감에 필요한 양일 수 있다. 유효량은 질병 진행이 없는 생존 기간을 연장시키거나, 전반적인 생존 기간을 증가시키거나, 암에 의한 이상 증상을 개선시킬 수 있다.In the formulation according to the present invention, the anti-Her2 antibody may be included in a therapeutically effective amount. "Effective amount" refers to an amount necessary or sufficient to bind to region II of the extracellular matrix domain of the HER2 receptor to inhibit the formation of dimers, thereby inhibiting receptor-mediated signaling pathways. For example, it is the amount necessary to prevent or treat various morphological symptoms of conditions associated with dimer formation of HER2. If the symptom is cancer, it may be an amount necessary for reducing the number of cancer cells, reducing the size of the tumor, inhibiting invasion of cancer cells, inhibiting tumor metastasis, inhibiting tumor growth, or reducing symptoms of cancer. An effective amount can prolong the survival period without disease progression, increase the overall survival period, or ameliorate adverse symptoms caused by cancer.
유효량은 피험자의 키, 무게, 질환의 유형 또는 중증도에 따라 다양할 수 있다. 유효량을 결정하고, 피험자에 투여될 수 있다. 분할된 용량 뿐 아니라, 교대 용량으로 매일 또는 연속적으로 투여될 수 있다. 용량은 치료 목적에 따라, 병용되는 약물 등에 따라 비율적으로 증가되거나 감소될 수 있다.The effective amount may vary depending on the subject's height, weight, type or severity of the disease. An effective amount can be determined and administered to a subject. As well as divided doses, it may be administered daily or continuously in alternating doses. The dose may be increased or decreased proportionally depending on the purpose of treatment, drugs used in combination, and the like.
"치료된", "치료하는" 또는 "치료"는 치료되는 상태, 장애 또는 질환과 연관되거나 이에 의해 유발되는 하나 이상의 증상의 감소 또는 경감을 포함한다. 예를 들어, 치료는 장애의 하나 이상의 증상을 감소시킬 수 있거나 장애를 완전히 제거할 수 있다.“Treated”, “treating” or “treatment” includes the reduction or relief of one or more symptoms associated with or caused by the condition, disorder or disease being treated. For example, treatment can reduce one or more symptoms of the disorder or can completely eliminate the disorder.
항-Her2 항체의 투여 수준은 환자에 대한 독성없이, 환자를 통해 목적하는 치료학적 반응에 효과적인 양으로 다양할 수 있다. 용량은 제형 내 항-Her2 항체의 활성, 투여 경로, 투여 시간, 치료 지속 시간, 병용하는 기타 약물 및/또는 화합물, 연령, 성별, 체중, 증상, 일반적인 건강 상태 및 치료될 환자의 이전의 의료 병력을 포함하는 다양한 인자를 통해 결정될 수 있다. The level of administration of the anti-Her2 antibody can vary in an amount effective for the desired therapeutic response through the patient, without toxicity to the patient. Dosage is the activity of the anti-Her2 antibody in the formulation, route of administration, time of administration, duration of treatment, other drugs and/or compounds in combination, age, sex, weight, symptoms, general health condition, and previous medical history of the patient to be treated. It can be determined through various factors including.
항-Her2 항체는 기타 Her2 타겟 항체 또는 화학요법제와 병용하여 투여될 수 있다. 병용 투여에는 별개의 제형 또는 단일 제약 제형을 사용하여 동시-투여하는 것과 순차적으로 투여하는 것이 포함된다. 기타 Her2 타겟 항체 또는 화학요법제는 항-Her2 항체 투여 전 또는 투여 후에 투여될 수 있다. 기타 Her2 타겟 항체 또는 화학요법제는 항-Her2 항체 투여와 동시에 투여될 수 있다. Anti-Her2 antibodies can be administered in combination with other Her2 target antibodies or chemotherapeutic agents. Concomitant administration includes co-administration and sequential administration using separate formulations or single pharmaceutical formulations. Other Her2 target antibodies or chemotherapeutic agents may be administered before or after administration of the anti-Her2 antibody. Other Her2 target antibodies or chemotherapeutic agents can be administered concurrently with administration of the anti-Her2 antibody.
기타 Her2 타겟 항체는 HER2와 결합하고, HER2를 과발현하는 암 세포의 성장을 억제시키는 항체일 수 있으며, 예를 들어, 트라스트주맙 (Herceptin®)일 수 있다. Other Her2 target antibodies may be antibodies that bind to HER2 and inhibit the growth of cancer cells that overexpress HER2, and may be, for example, trastzumab (Herceptin®).
화학요법제는 암의 치료에 유용한 화합물로, 예를 들어 안트라사이클린, 알킬화제 (예를 들어, 미토마이신 C), 알킬 술포네이트, 아지리딘, 에틸렌이민, 메틸멜라민, 질소 머스타드, 니트로소우레아, 항생제, 항대사물질, 엽산 유사체 (예를 들어, 디히드로폴레이트 환원효소 억제제, 예컨대 메토트렉세이트), 퓨린 유사체, 피리미딘 유사체, 효소, 포도필로톡신, 또는 백금-함유 제제 등을 포함한다. Chemotherapeutic agents are compounds useful in the treatment of cancer, such as anthracyclines, alkylating agents (e.g., mitomycin C), alkyl sulfonates, aziridines, ethylenimine, methylmelamine, nitrogen mustard, nitrosourea, antibiotics. , Antimetabolites, folic acid analogs (e.g., dihydrofolate reductase inhibitors such as methotrexate), purine analogs, pyrimidine analogs, enzymes, podophyllotoxin, or platinum-containing agents, and the like.
구체적으로, 부설판, 임프로설판, 피포설판, 벤조데파, 카르보쿠온, 메트우레데파, 우레데파, 알트레타민, 트리에틸렌멜라민, 트리에틸렌포스포르아미드, 트리에틸렌티오포스포르아미드, 트리메틸올로멜라민, 클로람부실, 클로르나파진, 시클로포스파미드, 에스트라무스틴, 이포스파미드, 메클로르에타민, 메클로르에타민 옥시드 히드로클로라이드, 멜팔란, 노벰비킨, 페네스테린, 프레드니무스틴, 트로포스파미드, 우라실 머스타드, 카무스틴, 클로로조토신, 포테무스틴, 로무스틴, 니무스틴, 라니무스틴, 다카바진, 만노무스틴, 미토브로니톨, 미토락톨, 피포브로만, 아클라시노마이신, 악티노마이신 F(1), 안트라마이신, 아자세린, 블레오마이신, 칵티노마이신, 카루비신, 카르지노필린, 크로모마이신, 닥티노마이신, 다우노루비신, 다우노마이신, 6-디아조-5-옥소-1-노르류신, 독소루비신, 에피루비신, 미토마이신 C, 미코페놀산, 노갈라마이신, 올리보마이신, 페플로마이신, 플리카마이신, 포르피로마이신, 퓨로마이신, 스트렙토니그린, 스트렙토조신, 투베르시딘, 우베니멕스, 지노스타틴, 조루비신, 데노프테린, 메토트렉세이트, 프테로프테린, 트리메트렉세이트, 플루다라빈, 6-머캅토퓨린, 티아미프린, 티오구아닌, 안시타빈, 아자시티딘, 6-아자우리딘, 카르모푸르, 시타라빈, 디데옥시우리딘, 독시플루리딘, 에노시타빈, 플록스우리딘, 플루오로우라실, 테가푸르, L-아스파라기나제, 풀모자임, 아세글라톤, 알도포스파미드 글리코시드, 아미노레불린산, 암사크린, 베스트라부실, 비스안트렌, 카르보플라틴, 시스플라틴, 데포파미드, 데메콜신, 디아지쿠온, 엘포르니틴, 엘립티늄 아세테이트, 에토글루시드, 에토포시드, 플루타미드, 질산갈륨, 히드록시우레아, 인터페론-알파, 인터페론-베타, 인터페론-감마, 인터류킨-2, 렌티난, 로니다민, 미토구아존, 미톡산트론, 모피다몰, 니트라크린, 펜토스타틴, 페나메트, 피라루비신, 포도필린산, 2-에틸히드라지드, 프로카르바진, 라족산, 시조피란, 스피로게르마늄, 파클리탁셀, 타목시펜, 테니포시드, 테누아존산, 트리아지쿠온, 2,2',2"-트리클로로트리에틸아민, 우레탄, 빈블라스틴, 빈크리스틴, 빈데신, 시클로스포린, 라파마이신 또는 아스코마이신, 또는 이의 유사체, 예를 들어 시클로스포린 A (CsA), 시클로스포린 G, FK-506, 코르티코스테로이드, 시클로포스파미드, 아자티오프린, 메토트렉세이트, 브레퀴나르, 레플루노미드, 미조리빈, 마이코페놀산, 마이코페놀레이트 모페틸, 15-데옥시스페르구알린, 또는 상기 화학요법제 중 2가지 이상의 병용 예를 들어 CHOP (시클로포스파미드, 독소루비신, 빈크리스틴 및 프레드니솔론의 병용) 또는 FOLFOX (5-FU 및 류코보빈과 병용된 옥살리플라틴) 등을 포함한다. Specifically, busulfan, improsulfan, pipesulfan, benzodepa, carboquone, meturedepa, uredepa, altretamine, triethylene melamine, triethylene phosphoramide, triethylene thiophosphoramide, trimethylol Romelamine, Chlorambucil, Chlornapazine, Cyclophosphamide, Estramustine, Ifosfamide, Mechlorethamine, Mechlorethamine Oxide Hydrochloride, Melphalan, Novemvicin, Penesterine, Prednimu Stein, Trophosphamide, Uracil Mustard, Carmustine, Chlorozotocin, Potemostin, Lomustine, Nimustine, Ranimostin, Takabazine, Mannomustine, Mitobronitol, Mitoractol, Pipobroman, Acla Sinomycin, Actinomycin F(1), Anthramycin, Azaserine, Bleomycin, Cactinomycin, Carubicin, Carginophylline, Chromomycin, Dactinomycin, Daunorubicin, Daunomycin, 6-dia Zo-5-oxo-1-norleucine, doxorubicin, epirubicin, mitomycin C, mycophenolic acid, nogalamycin, olivomycin, peplomycin, plicamycin, porpyromycin, puromycin, streptonigreen , Streptozosine, tubercidine, ubenimex, zinostatin, zorubicin, denopterin, methotrexate, pteropterin, trimetrexate, fludarabine, 6-mercaptopurine, thiamiprine, thioguanine , Ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxyfluridine, enocitabine, phloxuridine, fluorouracil, tegafur, L-aspa Laginase, fulmozyme, aceglatone, aldophosphamide glycoside, aminolevulinic acid, amsacrine, bestrabusil, bisanthrene, carboplatin, cisplatin, depopamide, demecolsin, diazicuone , Elfornitine, elliptinium acetate, etogluside, etoposide, flutamide, gallium nitrate, hydroxyurea, interferon-alpha, interferon-beta, interferon-gamma, interleukin-2, lentinan, ronidamine, Mitoguazone, mitoxantrone, furdamol, nitracrine, pentostatin, fenamet, pyrarubicin, podophyllic acid, 2-ethylhydrazide, procarbazine, razox acid, sizopyran, spirogermanium, paclitaxel, tamoxifen , Teniposide, tenuazonic acid, triazequon, 2,2',2"-trichlorotriethylamine, urethane, vinblastine, vincristine, vindesine, cyclosporine, rapamycin or ascomycin, or analogs thereof, For example, Cyclosporin A (CsA), Cyclosporin G, FK-506, Corticosteroids, Cyclophosphamide, Azathioprine, Methotrexate, Brequinar, Leflunomide, Mizoribine, Mycophenolic Acid, Mycophenol Rate mofetil, 15-deoxyspergualine, or a combination of two or more of the above chemotherapeutic agents, such as CHOP (a combination of cyclophosphamide, doxorubicin, vincristine and prednisolone) or FOLFOX (5-FU and leuco Oxaliplatin used in combination with bobbin), and the like.
항체 투여량은 일반적으로, 약 0.1 mg/kg 내지 약 50 mg/kg일 수 있다. 화학요법제를 투여하는 경우, 통상적으로 공지된 투여량으로 투여하거나, 화학요법제의 투여에 기인하는 부작용 또는 약물의 병용 작용으로 인해 더 낮은 양의 항체를 투여할 수 있다. 화학요법제에 대한 투여 스케쥴 및 제조는 제조업자의 지시에 따라 사용하거나, 또는 숙련자에 의해 실험적으로 결정할 수 있다. Antibody dosages can generally be from about 0.1 mg/kg to about 50 mg/kg. When administering a chemotherapeutic agent, it is usually administered at a known dosage, or a lower amount of the antibody may be administered due to side effects resulting from the administration of the chemotherapeutic agent or the combined action of drugs. The dosing schedule and preparation for the chemotherapeutic agent can be used according to the manufacturer's instructions, or can be determined experimentally by a skilled person.
약학 제형은 예를 들어 정맥내 투여, 근육내, 복강내, 뇌척수내, 피하, 관절내, 활막내, 수막강내, 경구, 국소, 또는 흡입 경로에 의해 환자에게 투여된다. 정맥내, 근육내 또는 피하 투여하는 것이 바람직하고, 본 발명의 약학 제형은 정맥내 투여될 수 있다. 피하 투여하는 경우 주사기; 주사 장치 (예: INJECT-EASE™ 및 GENJECT™ 장치); 주사기 펜 (예: GENPEN™); 바늘없는 장치 (예: MEDIJECTOR™ 및 BIOJECTOR™); 또는 피하 패치 전달 시스템을 사용할 수 있다. Pharmaceutical formulations are administered to a patient by, for example, intravenous administration, intramuscular, intraperitoneal, cerebrospinal, subcutaneous, intraarticular, intrasynovial, intrathecal, oral, topical, or inhalation routes. It is preferred to administer intravenously, intramuscularly or subcutaneously, and the pharmaceutical formulation of the present invention may be administered intravenously. Syringes for subcutaneous administration; Injection devices (eg INJECT-EASE™ and GENJECT™ devices); Syringe pens (eg GENPEN™); Needleless devices (eg MEDIJECTOR™ and BIOJECTOR™); Alternatively, a subcutaneous patch delivery system can be used.
또한, 본 발명의 약학 제형 및 이의 사용에 대한 지시사항을 제공하는 제조품을 제작할 수 있다. 제조품은 병, 바이알 (예: 이중 챔버 바이알), 주사기 (예: 이중 챔버 주사기) 또는 튜브 형태의 용기를 포함할 수 있다. 용기는 유리 또는 플라스틱으로 제조될 수 있다. 용기에 본 발명의 제형이 포함되고, 용기에 또는 용기에 부착된 라벨에 사용에 대한 지시사항을 표시할 수 있다. 제조품은 기타 완충제, 희석제, 충진제, 바늘, 주사기와 같은 추가 구성 및 사용에 대한 지시사항을 표시한 패키지 삽입물을 추가로 포함할 수 있다. In addition, it is possible to prepare an article of manufacture that provides the pharmaceutical formulation of the present invention and instructions for its use. Articles of manufacture may include bottles, vials (eg double chamber vials), syringes (eg double chamber syringes) or tubular containers. The container can be made of glass or plastic. The container contains the formulation of the present invention, and instructions for use may be indicated on the container or on a label affixed to the container. The article of manufacture may further include package inserts indicating instructions for further configuration and use, such as other buffers, diluents, fillers, needles, and syringes.
실시예Example
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 예시하기 위한 것으로서, 본 발명의 범위가 이들 실시예에 의해 제한되는 것으로 해석되지는 않는 것은 당업계에서 통상의 지식을 가진 자에게 있어서 자명할 것이다.Hereinafter, the present invention will be described in more detail through examples. These examples are for illustrative purposes only, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not construed as being limited by these examples.
실시예 1. pH 및 버퍼 스크리닝Example 1. pH and buffer screening
1-1. 퍼투주맙 제형의 제조1-1. Preparation of Pertuzumab Formulation
퍼투주맙 항체 제형을 표 1에서와 같이 제조하였다. 퍼투주맙 항체의 서열과 관련하여, US7862817B2 또는 US7537931B2에 기재된 구체적인 항체 서열을 참고할 수 있다. 퍼투주맙의 산성 변이체 관련하여서는 US8652474B2 또는 US9181346B2, 산성 변이체 또는 이황화 환원 변이체를 포함하는 퍼투주맙의 비결합 시스테인 변이체 (unpaired cysteine variants)에 대하여는 US9815904B2, US9969811B, 퍼투주맙의 VHS 연장 변이체 (VHS extension variants)에 대하여는 US7560111B2, US8241630B2 또는 US7879325B2를 참고할 수 있다. 퍼투주맙 항체를 36 mg/mL 이상으로 농축하고, 농축된 DS (drug substance)에 pH 4.0-5.0 아세테이트(아세트산염), pH 5.0-6.0 히스티딘-HCl, pH 6.0-7.0 포스페이트(인산염) 을 사용하여, 3시간, 5시간, 10시간 간격으로 3회에 나누어 2-8℃ 콜드 챔버 (cold chamber)에서 투석하였으며, pH 4.0-5.0 아세테이트(아세트산염), pH 5.0-6.0 히스티딘-HCl, pH 6.0-7.0 포스페이트(인산염) 각각으로 교환을 수행하였다. 목적하는 pH를 가지는 것으로 확인된 DS에 pH 4.0-5.0 아세테이트(아세트산염), pH 5.0-6.0 히스티딘-HCl, pH 6.0-7.0 포스페이트(인산염) 각각 및 20X 폴리솔베이트 20 용액 (0.4%)을 투입하여, 초기 DS 농도가 30 mg/mL, 폴리솔베이트 20이 0.02% 농도로 포함된 제형을 제조하였다. Pertuzumab antibody formulation was prepared as in Table 1. With regard to the sequence of the Pertuzumab antibody, reference may be made to the specific antibody sequence described in US7862817B2 or US7537931B2. Regarding the acidic variant of Pertuzumab, US8652474B2 or US9181346B2, the unpaired cysteine variants of Pertuzumab including the acidic variant or the reducing disulfide variant are US9815904B2, US9969811B, VHS extension variants of Pertuzumab (VHS extension variants). For more information, reference may be made to US7560111B2, US8241630B2 or US7879325B2. Pertuzumab antibody was concentrated to 36 mg/mL or more, and pH 4.0-5.0 acetate (acetate), pH 5.0-6.0 histidine-HCl, pH 6.0-7.0 phosphate (phosphate) was used in the concentrated DS (drug substance). , Divided into 3 times at 3, 5, and 10 hour intervals and dialyzed in a 2-8°C cold chamber, pH 4.0-5.0 acetate (acetate), pH 5.0-6.0 histidine-HCl, pH 6.0- Exchange was carried out with each of 7.0 phosphates (phosphates). In the DS confirmed to have the desired pH, pH 4.0-5.0 acetate (acetate), pH 5.0-6.0 histidine-HCl, pH 6.0-7.0 phosphate (phosphate), respectively, and 20X polysorbate 20 solution (0.4%) were added. Thus, a formulation containing an initial DS concentration of 30 mg/mL and polysorbate 20 at a concentration of 0.02% was prepared.
퍼투주맙 항체 30 mg/mL, 부형제로 수크로오스 120 mM, 0.02% PS20 계면활성제를 고정한 상태에서, pH 4.0-7.0 범위 중 0.5의 간격으로 설정한 pH 하에서 3 종류의 pH 4.0-5.0 아세테이트(아세트산염), pH 5.0-6.0 히스티딘-HCl, pH 6.0-7.0 포스페이트(인산염)을 이용하여, 퍼투주맙 항체에 적합한 제형을 스크리닝 하고자 하였다. 상기 3종류 성분 고유 특성에 따라 샘플의 pH 범위가 선정되었고, 샘플 3 및 샘플 4는 pH 5.0으로 동일하고, 샘플 6 및 샘플 7은 pH 6.0으로 동일하여 3 반복 실험하여 평균값을 기재하였다. 시판 중인 대조약 Perjeta®는, 히스티딘-아세테이트를 포함하고, 역시 3 반복 실험하여 평균값을 기재하였다. Pertuzumab Antibody 30 mg/mL, Sucrose 120 mM as an excipient, 0.02% PS20 Surfactant fixed, 3 types of pH 4.0-5.0 acetate (acetate) under a pH set at intervals of 0.5 in the range of pH 4.0-7.0 , pH 5.0-6.0 histidine-HCl, pH 6.0-7.0 phosphate (phosphate) was used to screen a formulation suitable for the Pertuzumab antibody. The pH range of the sample was selected according to the intrinsic characteristics of the three components, and Samples 3 and 4 were the same as pH 5.0, and Samples 6 and 7 were the same as pH 6.0, and the average value was described by repeating three experiments. The commercially available control drug Perjeta® contains histidine-acetate, and the average value was recorded by repeating three experiments.
Figure PCTKR2020015969-appb-T000001
Figure PCTKR2020015969-appb-T000001
1-2. 고온 스트레스 조건에서 SE-HPLC 분석 결과1-2. SE-HPLC analysis results under high temperature stress conditions
실시예 1-1에서 제조한 표 1의 샘플을 0.5mL씩 바이알에 필링 (filing)하였으며, 40℃±2℃ 안정성 챔버 (chamber)에 1주, 2주, 4주 샘플을 챔버-인 (chamber-in)하고, 각 타임 포인트에 샘플을 챔버-아웃 (chamber-out)하였다. SE-HPLC (Size Exclusion High Performance Liquid Chromatography: Tosoh Bioscience)를 사용하여 40℃±2℃ 초기, 1주, 2주 및 4주 조건에서 각 샘플에 대하여 HMW (High molecular weight species), 단량체 및 LMW (Low molecular weight species) 비율을 분석하였다. 분석 결과를 표 2 및 도 1 내지 도 3에 나타내었다. The sample of Table 1 prepared in Example 1-1 was filled into a vial by 0.5 mL, and samples were placed in a chamber for 1 week, 2 weeks, and 4 weeks in a 40°C±2°C stability chamber. -in), and chamber-out the sample at each time point. Using SE-HPLC (Size Exclusion High Performance Liquid Chromatography: Tosoh Bioscience), HMW (High molecular weight species), monomer and LMW (high molecular weight species), monomers, and LMW ( Low molecular weight species) ratio was analyzed. The analysis results are shown in Table 2 and FIGS. 1 to 3.
Figure PCTKR2020015969-appb-T000002
Figure PCTKR2020015969-appb-T000002
SE-HPLC 분석 결과 고온 스트레스 4주 조건 하에서 pH 5.0, 5.5, 6.0의 히스티딘-HCl을 포함하는 샘플 4 내지 샘플 6이 아세테이트 또는 포스페이트를 포함하는 샘플 1 내지 3 및 샘플 7 내지 9 보다 %HMW가 적어 더 안정함을 확인하였다. 또한, Perjeta®와 비교하여도 %HMW의 수준이 동등성 범위 또는 더 적어 그 안정성을 확인할 수 있었다. SE-HPLC analysis showed that Samples 4 to 6 containing histidine-HCl at pH 5.0, 5.5, and 6.0 under high temperature stress 4 weeks had less %HMW than Samples 1 to 3 and Samples 7 to 9 containing acetate or phosphate. It was confirmed that it is more stable. In addition, even compared to Perjeta®, the level of %HMW was in the range of equivalence or less, so its stability could be confirmed.
1-3. 결론1-3. conclusion
실시예 1-2에 따르면, pH 5.0-6.0의 히스티딘-HCl을 포함하는 조성물이 아세테이트 또는 포스페이트를 포함하는 조성물보다 안정성이 우수하고, Perjeta®와 비교하여도 동등 또는 그 이상의 안정성을 나타냄을 확인하였다. According to Example 1-2, it was confirmed that the composition containing histidine-HCl at pH 5.0-6.0 has better stability than the composition containing acetate or phosphate, and exhibits equivalent or higher stability compared to Perjeta®. .
실시예 2. 버퍼 및 안정화제 스크리닝Example 2. Buffer and stabilizer screening
2-1. 퍼투주맙 제형의 제조2-1. Preparation of Pertuzumab Formulation
실시예 1-1과 동일한 방법으로 표 3에 따른 퍼투주맙 제형을 제조하였다. 상기 실시예 1-2 분석 결과, %HMW 측면에서 효과가 좋았던 샘플 4 내지 6의 히스티딘-HCl 포함하는 제형을 더 스터디하고자, Perjeta®와 동일한 pH 6.0로 고정하고, 추가 성분으로서 소정 농도의 당, 당 알코올, 또는 제2 아미노산 포함하는 조성의 안정성을 비교하였다. 한편, 이들의 비교군으로 다른 조성은 동일하되 히스티딘-HCl을 숙시네이트(숙신산염)로 대체하여 그 결과를 비교하였다. 샘플 1, 샘플 6 및 Perjeta®는 숙시네이트, 히스티딘-HCl 및 히스티딘- 아세테이트를 포함하는 것을 제외하고는 동일 조성으로, 3 반복 실험 수행하여 그 평균값을 확인하였고, 본 실시예 2에서는 1주, 2주, 4주 및 8주까지의 실험 결과를 확인함으로써, 조금 더 장기 안정성을 확인하고자 하였다.A pertuzumab formulation according to Table 3 was prepared in the same manner as in Example 1-1. As a result of the analysis of Example 1-2, in order to further study the formulations containing histidine-HCl of Samples 4 to 6, which were effective in terms of %HMW, the same pH as Perjeta® was fixed to 6.0, and as an additional component, a predetermined concentration of sugar, The stability of the composition comprising a sugar alcohol or a second amino acid was compared. On the other hand, as a comparative group of these, the other composition was the same, but histidine-HCl was replaced with succinate (succinate), and the results were compared. Samples 1, 6, and Perjeta® were of the same composition except that succinate, histidine-HCl, and histidine-acetate were included, and the average value was confirmed by performing 3 repeated experiments. In this Example 2, 1 week, 2 By confirming the experimental results up to weeks, 4 weeks and 8 weeks, it was attempted to confirm a little more long-term stability.
Figure PCTKR2020015969-appb-T000003
Figure PCTKR2020015969-appb-T000003
2-2. 고온 스트레스 조건에서 SE-HPLC 분석 결과2-2. SE-HPLC analysis results under high temperature stress conditions
실시예 2-1에서 제조한 표 3의 샘플을 0.5mL씩 바이알에 필링 (filing)하였으며, 40℃±2℃ 안정성 챔버 (chamber)에서 1주, 2주, 4주, 8주 샘플을 각 타임 포인트에 챔버-아웃 (chamber-out)하였다. SE-HPLC를 사용하여 40℃±2℃ 초기, 1주, 2주, 4주 및 8주 조건에서 각 샘플에 대하여 HMW, 단량체 및 LMW 비율을 분석하였다. 분석 결과를 표 4 및 도 4 내지 도 6에 나타내었다. The sample of Table 3 prepared in Example 2-1 was filled into a vial with 0.5 mL each, and samples for 1 week, 2 weeks, 4 weeks, and 8 weeks in a 40°C±2°C stability chamber were each timed. Chamber-out at the point. Using SE-HPLC, HMW, monomer and LMW ratios were analyzed for each sample under the conditions of 40° C.±2° C. initial, 1 week, 2 weeks, 4 weeks and 8 weeks. The analysis results are shown in Table 4 and FIGS. 4 to 6.
Figure PCTKR2020015969-appb-T000004
Figure PCTKR2020015969-appb-T000004
SE-HPLC 분석 결과 고온 스트레스 8주 조건 하에서 숙시네이트를 포함한 샘플 1 내지 5에 비해 히스티딘-HCl을 포함하는 샘플 6 내지 10에서 %HMW가 적어 더 안정함을 확인하였다. As a result of SE-HPLC analysis, it was confirmed that the %HMW was less in the samples 6 to 10 containing histidine-HCl compared to the samples 1 to 5 containing succinate under the condition of 8 weeks of high temperature stress and thus was more stable.
그리고, 히스티딘-HCl과 함께 수크로오스, 트레할로스, 솔비톨, 알지닌 및 글리신을 포함하는 샘플 6 내지 10 모두 Perjeta®와 비교하여 %HMW가 적어 더 안정함을 확인하였다. In addition, it was confirmed that samples 6 to 10 containing sucrose, trehalose, sorbitol, arginine, and glycine along with histidine-HCl had less %HMW compared to Perjeta®, and thus were more stable.
2-3. 냉동-해동 5 사이클 조건에서 SE-HPLC 분석 결과2-3. SE-HPLC analysis results under freeze-thaw 5 cycle conditions
-70℃±10℃ 냉동-해동 5 사이클 조건에서 표 3의 각 샘플에 대하여, SE-HPLC 를 사용하여 HMW, 단량체, LMW 비율을 분석하였다. 분석 결과를 표 5, 도 7에 나타내었다.For each sample of Table 3 under conditions of -70°C±10°C freeze-thaw 5 cycles, the ratio of HMW, monomer, and LMW was analyzed using SE-HPLC. The analysis results are shown in Table 5 and Fig. 7.
Figure PCTKR2020015969-appb-T000005
Figure PCTKR2020015969-appb-T000005
SE-HPLC 분석 결과 -70℃±10℃ 냉동-해동 5 사이클 조건에서 숙시네이트를 포함한 샘플 1 내지 5에 비해 히스티딘-HCl을 포함하는 샘플 6 내지 10에서 %HMW가 적어 더 안정함을 확인하였다. As a result of SE-HPLC analysis, it was confirmed that %HMW was less in samples 6 to 10 containing histidine-HCl compared to samples 1 to 5 including succinate under conditions of -70°C±10°C freeze-thaw 5 cycles, and thus more stable.
그리고, 히스티딘-HCl과 함께 수크로오스, 트레할로스, 솔비톨, 알지닌 및 글리신을 포함하는 샘플 6 내지 10 모두 Perjeta®와 비교하여 %HMW가 적어 더 안정함을 확인하였다.In addition, it was confirmed that samples 6 to 10 containing sucrose, trehalose, sorbitol, arginine, and glycine along with histidine-HCl had less %HMW compared to Perjeta®, and thus were more stable.
2-4. 교반 조건에서 SE-HPLC 분석 결과2-4. SE-HPLC analysis results under stirring conditions
400rpm 교반 72시간 후 표 3의 각 샘플에 대하여, HMW, 단량체, LMW 비율을 SE-HPLC를 사용하여 분석하였다. 분석 결과를 표 6, 도 8에 나타내었다.For each sample in Table 3 after 72 hours of stirring at 400 rpm, the ratio of HMW, monomer, and LMW was analyzed using SE-HPLC. The analysis results are shown in Table 6 and FIG. 8.
Figure PCTKR2020015969-appb-T000006
Figure PCTKR2020015969-appb-T000006
SE-HPLC 분석 결과 400rpm 교반 72시간 후 숙시네이트를 포함한 샘플 1 내지 5에 비해 히스티딘-HCl을 포함하는 샘플 6 내지 10에서 %HMW가 적어 더 안정함을 확인하였다. As a result of SE-HPLC analysis, after 72 hours of stirring at 400 rpm, it was confirmed that the %HMW was less in the samples 6 to 10 containing histidine-HCl compared to the samples 1 to 5 containing succinate and thus more stable.
그리고, 히스티딘-HCl과 함께 수크로오스, 트레할로스, 솔비톨, 알지닌 및 글리신을 포함하는 샘플 6 내지 10 모두 Perjeta®와 비교하여 %HMW가 적어 더 안정함을 확인하였다.In addition, it was confirmed that samples 6 to 10 containing sucrose, trehalose, sorbitol, arginine, and glycine along with histidine-HCl had less %HMW compared to Perjeta®, and thus were more stable.
2-5. 결론2-5. conclusion
실시예 2-2 내지 실시예 2-4에 따르면, pH 6.0 히스티딘-HCl을 포함하는 제형이 숙시네이트를 포함하는 제형보다 안정성이 우수하고, Perjeta®와 비교하여도 동등 또는 그 이상의 안정성을 나타냄을 확인하였다.According to Examples 2-2 to 2-4, it was found that the formulation containing pH 6.0 histidine-HCl has better stability than the formulation containing succinate and exhibits equivalent or higher stability compared to Perjeta®. Confirmed.
나아가 히스티딘-HCl과 함께 수크로오스, 트레할로스, 솔비톨, 알지닌 및 글리신을 포함하는 제형 모두 Perjeta®와 비교하여도 동등 또는 그 이상의 안정성을 나타냄을 확인하였다.Furthermore, it was confirmed that all formulations containing sucrose, trehalose, sorbitol, arginine, and glycine along with histidine-HCl showed equal or higher stability compared to Perjeta®.
실시예 3. 최적의 pH 및 안정화제 확인Example 3. Identification of optimal pH and stabilizer
3-1. 퍼투주맙 제형의 제조3-1. Preparation of Pertuzumab Formulation
실시예 1-1과 동일한 방법으로 표 7에 따른 제형을 제조하였다. 상기 실시예 2에서 효과가 좋았던 추가 성분 중에서 수크로스, 솔비톨, 및 글리신의 농도 변화를 주고, 거기에 제3의 아미노산인 메티오닌을 추가하여, 제형 안정성을 확인하고자 하였다. 샘플들은 3 반복 실험하여 평균 값으로 결과를 확인하였고, 실시예 2의 Perjeta® 실험값과 비교하였다.A formulation according to Table 7 was prepared in the same manner as in Example 1-1. Among the additional ingredients having good effects in Example 2, the concentration of sucrose, sorbitol, and glycine was changed, and methionine, a third amino acid, was added thereto to confirm the formulation stability. The samples were subjected to 3 repeated experiments to confirm the result as an average value, and compared with the Perjeta® experimental value of Example 2.
Figure PCTKR2020015969-appb-T000007
Figure PCTKR2020015969-appb-T000007
20 mM 히스티딘-HCl을 포함하는 샘플에서 pH를 각각 5.5, 6.0 및 6.5로 조절하기 위해 포함되는 L-히스티딘 및 L-히스티딘 모노하이드로클로라이드 일수화물의 농도는 표 8과 같다. Table 8 shows the concentrations of L-histidine and L-histidine monohydrochloride monohydrate contained in the sample containing 20 mM histidine-HCl to adjust the pH to 5.5, 6.0, and 6.5, respectively.
Figure PCTKR2020015969-appb-T000008
Figure PCTKR2020015969-appb-T000008
3-2. 고온 스트레스 조건에서 SE-HPLC 분석 결과3-2. SE-HPLC analysis results under high temperature stress conditions
실시예 3-1에서 제조한 표 7의 샘플을 0.5mL씩 바이알에 필링 (filing)하였으며, 40℃±2℃ 안정성 챔버 (chamber)에서 1주, 2주, 4주, 8주 샘플을 각 타임 포인트에 챔버-아웃 (chamber-out)하였다. SE-HPLC를 사용하여 40℃±2℃ 초기, 1주, 2주, 4주 및 8주 조건에서 각 샘플에 대하여 HMW, 단량체 및 LMW 비율을 분석하였다. 분석 결과를 표 9, 및 도 9 내지 도 11에 나타내었다.The sample of Table 7 prepared in Example 3-1 was filled into a vial with 0.5 mL each, and samples for 1 week, 2 weeks, 4 weeks, and 8 weeks were each time in a 40°C±2°C stability chamber. Chamber-out at the point. Using SE-HPLC, HMW, monomer and LMW ratios were analyzed for each sample under the conditions of 40° C.±2° C. initial, 1 week, 2 weeks, 4 weeks and 8 weeks. The analysis results are shown in Table 9 and FIGS. 9 to 11.
Figure PCTKR2020015969-appb-T000009
Figure PCTKR2020015969-appb-T000009
SE-HPLC 분석 결과 고온 스트레스 8주 조건 하에서 히스티딘-HCl에 수크로오스, 솔비톨 또는 글리신과 메티오닌을 포함하는 샘플 1 내지 샘플 15의 제형의 경우 Perjeta®와 비교하여 %HMW가 적어 더 안정함을 확인하였다. As a result of SE-HPLC analysis, it was confirmed that the formulations of Samples 1 to 15 containing sucrose, sorbitol, or glycine and methionine in histidine-HCl under the condition of high temperature stress for 8 weeks had less %HMW compared to Perjeta® and thus more stable.
3-3. 냉동-해동 5 사이클 조건에서 SE-HPLC 분석 결과3-3. SE-HPLC analysis results under freeze-thaw 5 cycle conditions
-70℃±10℃ 냉동-해동 5 사이클 조건에서 표 7의 각 샘플에 대하여, SE-HPLC 를 사용하여 HMW, 단량체, 및 LMW 비율을 분석하였다. 분석 결과를 표 10, 도 12에 나타내었다. For each sample in Table 7 under conditions of -70°C±10°C freeze-thaw 5 cycles, the HMW, monomer, and LMW ratios were analyzed using SE-HPLC. The analysis results are shown in Tables 10 and 12.
Figure PCTKR2020015969-appb-T000010
Figure PCTKR2020015969-appb-T000010
SE-HPLC 분석 결과 -70℃±10℃ 냉동-해동 5 사이클 조건에서 수크로오스 또는 솔비톨과 메티오닌을 포함하는 샘플 1 내지 샘플 10 제형의 경우 Perjeta®와 비교하여 % HMW가 적어 더 안정함을 확인하였다. As a result of SE-HPLC analysis, it was confirmed that the formulations of Samples 1 to 10 containing sucrose or sorbitol and methionine under the conditions of -70°C±10°C freeze-thaw 5 cycles were more stable due to less% HMW compared to Perjeta®.
3-4. 교반 조건에서 SE-HPLC 분석 결과3-4. SE-HPLC analysis results under stirring conditions
400rpm 교반 72시간 후 표 7의 각 샘플에 대하여, HMW, 단량체, 및 LMW 비율을 SE-HPLC를 사용하여 분석하였다. 분석 결과를 표 11, 도 13에 나타내었다.For each sample in Table 7 after 72 hours of stirring at 400 rpm, the ratio of HMW, monomer, and LMW was analyzed using SE-HPLC. The analysis results are shown in Table 11 and FIG. 13.
Figure PCTKR2020015969-appb-T000011
Figure PCTKR2020015969-appb-T000011
SE-HPLC 분석 결과 400rpm 교반 72시간 후 조건 하에서 히스티딘-HCl에 수크로오스, 솔비톨 또는 글리신과 메티오닌을 포함하는 샘플 1 내지 샘플 15의 제형의 경우 Perjeta®와 비교하여 % HMW가 적어 더 안정함을 확인하였다. As a result of SE-HPLC analysis, it was confirmed that the formulations of Samples 1 to 15 containing sucrose, sorbitol, or glycine and methionine in histidine-HCl under conditions after 72 hours of stirring at 400 rpm had less% HMW compared to Perjeta® and thus more stable. .
3-5. 결론3-5. conclusion
실시예 3-2 내지 실시예 3-4에 따르면, 히스티딘-HCl에 수크로오스 또는 솔비톨 과 메티오닌을 포함하는 샘플 1 내지 샘플 10의 제형의 경우, 고온 스트레스, 냉동/해동 및 교반 조건에서 Perjeta®와 비교하여 % HMW가 적어 더 안정함을 확인하였다.According to Examples 3-2 to 3-4, the formulations of Samples 1 to 10 containing sucrose or sorbitol and methionine in histidine-HCl were compared with Perjeta® under high temperature stress, freezing/thawing and stirring conditions. Thus, it was confirmed that the% HMW was less and more stable.
한편, 수크로오스, 솔비톨 또는 글리신의 농도 변화에 따른 %HMW 차이는 크지 않은 것을 확인하였다. 다만, pH 5.5 및 6.0의 샘플의 경우, pH 6.5의 샘플과 비교하여 % HMW가 적어 더 안정함을 확인하였다. On the other hand, it was confirmed that the %HMW difference according to the change in the concentration of sucrose, sorbitol or glycine was not large. However, in the case of the samples of pH 5.5 and 6.0, it was confirmed that the% HMW was less than that of the samples of pH 6.5 and thus more stable.
실시예 4. 2종 이상의 아미노산의 효과 분석 Example 4. Analysis of the effect of two or more amino acids
4-1. 비교 분석 제형의 선정4-1. Selection of formulations for comparative analysis
본 발명의 약제학적 조성물의 2종 이상의 아미노산의 효과를 확인하기 위하여 하기 표 12에서 보는 바와 같이, 실시예 2의 샘플 6, 8, 및 10과 실시예 3의 샘플 3, 8 및 13을 실시예 2의 Perjeta®와 HMW, 단량체 및 LMW 비율을 비교 분석하여 유의적인 결과가 도출되는지 확인하고자 하였다. In order to confirm the effect of two or more amino acids of the pharmaceutical composition of the present invention, as shown in Table 12 below, Samples 6, 8, and 10 of Example 2 and Samples 3, 8 and 13 of Example 3 were used as examples. 2, Perjeta® and the ratio of HMW, monomer and LMW were compared and analyzed to confirm whether significant results were obtained.
Figure PCTKR2020015969-appb-T000012
Figure PCTKR2020015969-appb-T000012
4-2. 고온 스트레스 조건에서 SE-HPLC 분석 결과4-2. SE-HPLC analysis results under high temperature stress conditions
SE-HPLC를 사용하여 40℃±2℃ 초기, 1주, 2주, 4주 및 8주 조건에서 표 12의 각 샘플에 대하여 HMW, 단량체 및 LMW 비율을 분석하였다. 분석 결과를 표 13 및 도 14 내지 16에 나타내었다.Using SE-HPLC, HMW, monomer and LMW ratios were analyzed for each sample of Table 12 under the conditions of 40°C±2°C initial, 1 week, 2 weeks, 4 weeks and 8 weeks. The analysis results are shown in Table 13 and FIGS. 14 to 16.
Figure PCTKR2020015969-appb-T000013
Figure PCTKR2020015969-appb-T000013
SE-HPLC 분석 결과 고온 스트레스 8주 조건 하에서 본 발명의 약제학적 조성물이 실시예 2의 Perjeta®와 비교하여 %HMW가 적어 더 안정함을 확인하였다.As a result of SE-HPLC analysis, it was confirmed that the pharmaceutical composition of the present invention was more stable due to less %HMW compared to Perjeta® of Example 2 under the condition of 8 weeks of high temperature stress.
그리고, 그 중에서도, 1종의 아미노산을 포함한 샘플 6(Example 2) 및 8(Example 2)보다, 2종 이상의 아미노산을 포함한 샘플 10(Example 2), 샘플 3(Example 3), 샘플 8(Example 3) 및 샘플 13(Example 3)가 %HMW가 적어 더 안정함을 확인하였으며, 그 중에서도 2종의 아미노산을 포함한 샘플 10(Example 2), 샘플 3(Example 3) 및 샘플 8(Example 3)보다 3종의 아미노산을 포함한 샘플 13(Example 3)이 가장 %HMW가 적어 가장 안정함을 확인하였다.And, among others, than Samples 6 (Example 2) and 8 (Example 2) containing one amino acid, Sample 10 (Example 2), Sample 3 (Example 3), and Sample 8 (Example 3) containing two or more amino acids. ) And Sample 13 (Example 3) were confirmed to be more stable due to less %HMW, and among them, 3 than Sample 10 (Example 2), Sample 3 (Example 3) and Sample 8 (Example 3) containing two kinds of amino acids. It was confirmed that Sample 13 (Example 3) containing species of amino acids had the least %HMW and was the most stable.
4-3. 교반 조건에서 SE-HPLC 분석 결과4-3. SE-HPLC analysis results under stirring conditions
400rpm 교반 72시간 후 표 12의 각 샘플에 대하여, HMW, 단량체, LMW 비율을 SE-HPLC를 사용하여 분석하였다. 분석 결과를 표 14 및 도 17에 나타내었다.For each sample in Table 12 after 72 hours of stirring at 400 rpm, the ratio of HMW, monomer, and LMW was analyzed using SE-HPLC. The analysis results are shown in Table 14 and FIG. 17.
Figure PCTKR2020015969-appb-T000014
Figure PCTKR2020015969-appb-T000014
SE-HPLC 분석 결과 400rpm 교반 72시간 후 본 발명의 약제학적 조성물이 실시예 2의 Perjeta®와 비교하여 %HMW가 적어 더 안정함을 확인하였다.As a result of SE-HPLC analysis, it was confirmed that the pharmaceutical composition of the present invention after 72 hours of stirring at 400 rpm had less %HMW compared to the Perjeta® of Example 2 and thus was more stable.
그리고, 그 중에서도, 1종의 아미노산을 포함한 샘플 6(Example 2) 및 8(Example 2)보다, 2종 이상의 아미노산을 포함한 샘플 10(Example 2), 샘플 3(Example 3), 샘플 8(Example 3) 및 샘플 13(Example 3)가 %HMW가 적어 더 안정함을 확인하였으며, 그 중에서도 2종의 아미노산을 포함한 샘플 10(Example 2), 샘플 3(Example 3) 및 샘플 8(Example 3)보다 3종의 아미노산을 포함한 샘플 13(Example 3)이 가장 %HMW가 적어 가장 안정함을 확인하였다.And, among others, than Samples 6 (Example 2) and 8 (Example 2) containing one amino acid, Sample 10 (Example 2), Sample 3 (Example 3), and Sample 8 (Example 3) containing two or more amino acids. ) And Sample 13 (Example 3) were confirmed to be more stable due to less %HMW, and among them, 3 than Sample 10 (Example 2), Sample 3 (Example 3) and Sample 8 (Example 3) containing two kinds of amino acids. It was confirmed that Sample 13 (Example 3) containing species of amino acids had the least %HMW and was the most stable.
4-4. 결론4-4. conclusion
실시예 4-2 및 실시예 4-3에 따르면, 본 발명의 약제학적 조성물이 실시예 2의 Perjeta®와 비교하여 %HMW가 적어 더 안정함을 확인하였다. According to Examples 4-2 and 4-3, it was confirmed that the pharmaceutical composition of the present invention was more stable due to less %HMW compared to Perjeta® of Example 2.
그리고, 1종의 아미노산을 포함한 제형보다 2종 이상의 아미노산을 포함한 제형이 더 안정하고, 그 중에서도 2종의 아미노산을 포함한 제형보다 3종의 아미노산을 포함한 것이 더 안정함을 확인하였다. In addition, it was confirmed that the formulation containing two or more amino acids was more stable than the formulation containing one amino acid, and among them, the formulation containing three amino acids was more stable than the formulation containing two amino acids.
이상으로 본 발명의 내용의 특정한 부분을 상세히 기술하였는바, 당업계의 통상의 지식을 가진 자에게 있어서, 이러한 구체적 기술은 단지 바람직한 실시양태일 뿐이며, 이에 의해 본 발명의 범위가 제한되는 것이 아닌 점은 명백할 것이다. 따라서, 본 발명의 실질적인 범위는 첨부된 청구항들과 그것들의 등가물에 의하여 정의된다고 할 것이다.As described above, a specific part of the content of the present invention has been described in detail, and for those of ordinary skill in the art, this specific technology is only a preferred embodiment, and the scope of the present invention is not limited thereto. It will be obvious. Accordingly, it will be said that the substantial scope of the present invention is defined by the appended claims and their equivalents.

Claims (42)

  1. 항-Her2 항체; 및 Anti-Her2 antibody; And
    2종 이상의 아미노산을 포함하고, It contains two or more kinds of amino acids,
    아세트산염을 포함하지 않는 약제학적 조성물.A pharmaceutical composition that does not contain acetate.
  2. 제1항에 있어서, 히스티딘, 메티오닌, 시스테인, 알지닌, 글리신, 프롤린, 라이신, 글루탐산, 이소루이신, 페닐알라닌, 티로신, 아스파트산으로 구성된 군에서 선택되는 2종 이상의 아미노산을 포함하는 약제학적 조성물.The pharmaceutical composition according to claim 1, comprising two or more amino acids selected from the group consisting of histidine, methionine, cysteine, arginine, glycine, proline, lysine, glutamic acid, isoleucine, phenylalanine, tyrosine, and aspartic acid. .
  3. 제1항 또는 제2항에 있어서, 히스티딘 및 히스티딘 이외의 아미노산을 포함하는 약제학적 조성물.The pharmaceutical composition according to claim 1 or 2, comprising histidine and an amino acid other than histidine.
  4. 제1항 내지 제3항 중 어느 한 항에 있어서, i) 히스티딘 및 ii) 메티오닌, 시스테인, 알지닌, 글리신, 프롤린, 라이신, 글루탐산, 이소루이신, 페닐알라닌, 티로신, 아스파트산으로 구성된 군에서 선택되는 1종 이상의 아미노산을 포함하는 약제학적 조성물.The method according to any one of claims 1 to 3, in the group consisting of i) histidine and ii) methionine, cysteine, arginine, glycine, proline, lysine, glutamic acid, isoleucine, phenylalanine, tyrosine, and aspartic acid. A pharmaceutical composition comprising one or more selected amino acids.
  5. 제1항 내지 제4항 중 어느 한 항에 있어서, 히스티딘 클로라이드 (histidine chloride), 히스티딘 포스페이트 (histidine phosphate), 또는 히스티딘 설페이트 (histidine sulfate)를 포함하는 약제학적 조성물.The pharmaceutical composition according to any one of claims 1 to 4, comprising histidine chloride, histidine phosphate, or histidine sulfate.
  6. 제1항 내지 제5항 중 어느 한 항에 있어서, 히스티딘 및 히스티딘 모노하이드로클로라이드 일수화물 (histidine monohydrochloride monohydrate)을 포함하는 약제학적 조성물.The pharmaceutical composition according to any one of claims 1 to 5, comprising histidine and histidine monohydrochloride monohydrate.
  7. 제1항 내지 제6항 중 어느 한 항에 있어서, 히스티딘 아세테이트를 포함하지 않는 약제학적 조성물.The pharmaceutical composition according to any one of claims 1 to 6, which does not comprise histidine acetate.
  8. 제1항 내지 제7항 중 어느 한 항에 있어서, 글루코스, 수크로오스, 트레할로스, 락토오스, 프럭토오스, 말토오스, 덱스트란, 글리세린, 에리트리톨, 아라비톨, 자일리톨, 솔비톨, 만니톨, 멜리바이오스, 멜레치토스, 라피노오스, 만노트리오스, 스타키오스, 말토오스, 락툴로오스, 말툴로오스, 글루시톨, 말티톨, 락티톨, 이소-말툴로오스로 구성된 군에서 선택되는 하나 이상의 당 또는 당알코올을 더 포함하는 약제학적 조성물.The method according to any one of claims 1 to 7, wherein glucose, sucrose, trehalose, lactose, fructose, maltose, dextran, glycerin, erythritol, arabitol, xylitol, sorbitol, mannitol, melibiose, meletitol , Raffinose, mannotrios, stachyose, maltose, lactulose, maltulose, glucitol, maltitol, lactitol, iso-maltulose, one or more sugars or sugar alcohols selected from the group consisting of Pharmaceutical composition comprising.
  9. 제1항 내지 제8항 중 어느 한 항에 있어서, 상기 당 또는 당 알코올은 수크로오스, 솔비톨 또는 트레할로스인 약제학적 조성물.The pharmaceutical composition according to any one of claims 1 to 8, wherein the sugar or sugar alcohol is sucrose, sorbitol or trehalose.
  10. 제1항 내지 제9항 중 어느 한 항에 있어서, 상기 히스티딘 이외의 아미노산, 또는 당 또는 당 알코올을 50 mM 내지 150 mM의 농도로 포함하는 약제학적 조성물.The pharmaceutical composition according to any one of claims 1 to 9, comprising an amino acid other than histidine, or a sugar or sugar alcohol at a concentration of 50 mM to 150 mM.
  11. 제1항 내지 제9항 중 어느 한 항에 있어서, pH 5.5 내지 6.0인 약제학적 조성물.The pharmaceutical composition according to any one of claims 1 to 9, which has a pH of 5.5 to 6.0.
  12. 제1항 내지 제10항 중 어느 한 항에 있어서, 상기 히스티딘 이외의 아미노산, 또는 당 또는 당 알코올: 항-Her2 항체의 몰비가 300 : 1 내지 450:1 또는 540:1 내지 690:1인 약제학적 조성물.The drug according to any one of claims 1 to 10, wherein the molar ratio of amino acids other than histidine or sugar or sugar alcohol: anti-Her2 antibody is 300:1 to 450:1 or 540:1 to 690:1. Composition.
  13. 제1항 내지 제11항 중 어느 한 항에 있어서, 히스티딘 : 히스티딘 모노하이드로클로라이드 일수화물을 0.2 내지 3.0:1의 몰비로 포함하는 약제학적 조성물.The pharmaceutical composition according to any one of claims 1 to 11, comprising histidine:histidine monohydrochloride monohydrate in a molar ratio of 0.2 to 3.0:1.
  14. 제1항 내지 제12항 중 어느 한 항에 있어서, 히스티딘 이외의 아미노산을 5 mM 내지 30 mM 또는 50 mM 내지 150 mM의 농도로 포함하는 약제학적 조성물.The pharmaceutical composition according to any one of claims 1 to 12, comprising an amino acid other than histidine in a concentration of 5 mM to 30 mM or 50 mM to 150 mM.
  15. 제1항 내지 제13항 중 어느 한 항에 있어서, 상기 항-Her2 항체 : 히스티딘 이외의 아미노산의 몰비는 1:10 내지 1:100, 300 : 1 내지 450:1 또는 500:1 내지 700:1인 약제학적 조성물.The method according to any one of claims 1 to 13, wherein the molar ratio of the anti-Her2 antibody: amino acids other than histidine is 1:10 to 1:100, 300:1 to 450:1, or 500:1 to 700:1 Phosphorus pharmaceutical composition.
  16. 제1항 내지 제14항 중 어느 한 항에 있어서, 상기 항-Her2 항체는 퍼투주맙(pertuzumab)인 것인 약제학적 조성물.The pharmaceutical composition according to any one of claims 1 to 14, wherein the anti-Her2 antibody is pertuzumab.
  17. 제1항 내지 제15항 중 어느 한 항에 있어서, 상기 항-Her2 항체를 20 mg/ml 내지 150mg/ml의 농도로 포함하는 약제학적 조성물.The pharmaceutical composition according to any one of claims 1 to 15, comprising the anti-Her2 antibody at a concentration of 20 mg/ml to 150 mg/ml.
  18. 제1항 내지 제16항 중 어느 한 항에 있어서, 20 mg/ml 내지 30mg/ml의 농도로 항-Her2 항체를 포함하는 IV 투여용 약제학적 조성물.The pharmaceutical composition for IV administration according to any one of claims 1 to 16, comprising an anti-Her2 antibody at a concentration of 20 mg/ml to 30 mg/ml.
  19. 제1항 내지 제17항 중 어느 한 항에 있어서, 폴리솔베이트 20 또는 폴리솔베이트 80을 추가로 포함하는 약제학적 조성물.18. The pharmaceutical composition according to any one of claims 1 to 17, further comprising polysorbate 20 or polysorbate 80.
  20. 항-Her2 항체 및 2종 이상의 아미노산을 포함하고, It contains an anti-Her2 antibody and two or more amino acids,
    아세트산염을 포함하지 않으며, Does not contain acetate,
    고온 보관 조건, 냉동/해동 조건 또는 교반 조건에서 안정성이 유지 또는 향상된 약제학적 조성물.A pharmaceutical composition that maintains or improves stability under high temperature storage conditions, freezing/thawing conditions, or stirring conditions.
  21. 제20항에 있어서, 40℃±2℃ 1주 이상의 보관 조건에서 크기 배제 HPLC(SE-HPLC)를 통해 측정된 HMW가 2% 이하인 약제학적 조성물.The pharmaceutical composition according to claim 20, wherein the HMW measured by size exclusion HPLC (SE-HPLC) at 40° C.±2° C. for 1 week or more is 2% or less.
  22. 제20항 또는 제21항에 있어서, -70℃±10℃ 냉동-해동 5 사이클 조건에서 크기 배제 HPLC(SE-HPLC)를 통해 측정된 HMW가 2% 이하인 약제학적 조성물.The pharmaceutical composition according to claim 20 or 21, wherein the HMW measured by size exclusion HPLC (SE-HPLC) under 5 cycles of -70°C±10°C freeze-thaw is 2% or less.
  23. 제20항 내지 제22항 중 어느 한 항에 있어서, 400rpm 교반 72시간 후 조건에서 크기 배제 HPLC(SE-HPLC)를 통해 측정된 HMW가 2% 이하인 약제학적 조성물.The pharmaceutical composition according to any one of claims 20 to 22, wherein the HMW measured by size exclusion HPLC (SE-HPLC) under the condition after 72 hours of stirring at 400 rpm is 2% or less.
  24. 제20항 내지 제23항 중 어느 한 항에 있어서, 당 또는 당 알코올을 추가로 포함하는 약제학적 조성물.The pharmaceutical composition according to any one of claims 20 to 23, further comprising a sugar or a sugar alcohol.
  25. 제20항 내지 제24항 중 어느 한 항에 있어서, 히스티딘, 메티오닌, 시스테인, 알지닌, 글리신, 프롤린, 라이신, 글루탐산, 이소루이신, 페닐알라닌, 티로신, 아스파트산으로 구성된 군에서 선택되는 2종 이상의 아미노산을 포함하는 약제학적 조성물.The method according to any one of claims 20 to 24, two selected from the group consisting of histidine, methionine, cysteine, arginine, glycine, proline, lysine, glutamic acid, isoleucine, phenylalanine, tyrosine, and aspartic acid. A pharmaceutical composition comprising the above amino acids.
  26. 제20항 내지 제25항 중 어느 한 항에 있어서, 히스티딘 및 히스티딘 이외의 아미노산을 포함하는 약제학적 조성물.26. A pharmaceutical composition according to any one of claims 20 to 25 comprising histidine and an amino acid other than histidine.
  27. 제20항 내지 제26항 중 어느 한 항에 있어서, i) 히스티딘 및 ii) 메티오닌, 시스테인, 알지닌, 글리신, 프롤린, 라이신, 글루탐산, 이소루이신, 페닐알라닌, 티로신, 아스파트산으로 구성된 군에서 선택되는 1종 이상의 아미노산을 포함하는 약제학적 조성물.The method according to any one of claims 20 to 26, in the group consisting of i) histidine and ii) methionine, cysteine, arginine, glycine, proline, lysine, glutamic acid, isoleucine, phenylalanine, tyrosine, aspartic acid. A pharmaceutical composition comprising one or more selected amino acids.
  28. 제20항 내지 제27항 중 어느 한 항에 있어서, 히스티딘 클로라이드 (histidine chloride), 히스티딘 포스페이트 (histidine phosphate), 또는 히스티딘 설페이트 (histidine sulfate)를 포함하는 약제학적 조성물.The pharmaceutical composition according to any one of claims 20 to 27, comprising histidine chloride, histidine phosphate, or histidine sulfate.
  29. 제20항 내지 제28항 중 어느 한 항에 있어서, 히스티딘 및 히스티딘 모노하이드로클로라이드 일수화물 (histidine monohydrochloride monohydrate)을 포함하는 약제학적 조성물.29. A pharmaceutical composition according to any one of claims 20 to 28 comprising histidine and histidine monohydrochloride monohydrate.
  30. 제20항 내지 제29항 중 어느 한 항에 있어서, 히스티딘 아세테이트를 포함하지 않는 약제학적 조성물.The pharmaceutical composition according to any one of claims 20 to 29, which does not comprise histidine acetate.
  31. 제20항 내지 제30항 중 어느 한 항에 있어서, 상기 당 또는 당 알코올은 글루코스, 수크로오스, 트레할로스, 락토오스, 프럭토오스, 말토오스, 덱스트란, 글리세린, 에리트리톨, 아라비톨, 자일리톨, 솔비톨, 만니톨, 멜리바이오스, 멜레치토스, 라피노오스, 만노트리오스, 스타키오스, 말토오스, 락툴로오스, 말툴로오스, 글루시톨, 말티톨, 락티톨, 이소-말툴로오스로 구성된 군에서 선택되는 하나 이상인 약제학적 조성물.The method according to any one of claims 20 to 30, wherein the sugar or sugar alcohol is glucose, sucrose, trehalose, lactose, fructose, maltose, dextran, glycerin, erythritol, arabitol, xylitol, sorbitol, mannitol. , Melibiose, meletithose, raffinose, mannotiose, stachyose, maltose, lactulose, maltulose, glucitol, maltitol, lactitol, iso-maltulose Pharmaceutical composition.
  32. 제20항 내지 제31항 중 어느 한 항에 있어서, 상기 당 또는 당 알코올은 수크로오스, 솔비톨 또는 트레할로스인 약제학적 조성물.The pharmaceutical composition according to any one of claims 20 to 31, wherein the sugar or sugar alcohol is sucrose, sorbitol or trehalose.
  33. 제20항 내지 제32항 중 어느 한 항에 있어서, 상기 히스티딘 이외의 아미노산, 또는 당 또는 당 알코올을 50 mM 내지 150 mM의 농도로 포함하는 약제학적 조성물.The pharmaceutical composition according to any one of claims 20 to 32, comprising an amino acid other than histidine, or a sugar or sugar alcohol at a concentration of 50 mM to 150 mM.
  34. 제20항 내지 제33항 중 어느 한 항에 있어서, 상기 히스티딘 이외의 아미노산, 또는 당 또는 당 알코올: 항-Her2 항체의 몰비가 300 : 1 내지 450:1 또는 540:1 내지 690:1인 약제학적 조성물.The drug according to any one of claims 20 to 33, wherein the molar ratio of amino acids other than histidine, or sugar or sugar alcohol: anti-Her2 antibody is 300:1 to 450:1 or 540:1 to 690:1. Composition.
  35. 제20항 내지 제34항 중 어느 한 항에 있어서, pH 5.5 내지 6.0인 약제학적 조성물.The pharmaceutical composition according to any one of claims 20 to 34, wherein the pH is 5.5 to 6.0.
  36. 제20항 내지 제35항 중 어느 한 항에 있어서, 히스티딘 : 히스티딘 모노하이드로클로라이드 일수화물을 0.2 내지 3.0:1의 몰비로 포함하는 약제학적 조성물.36. A pharmaceutical composition according to any one of claims 20 to 35, comprising histidine:histidine monohydrochloride monohydrate in a molar ratio of 0.2 to 3.0:1.
  37. 제20항 내지 제36항 중 어느 한 항에 있어서, 히스티딘 이외의 아미노산을 5 mM 내지 30 mM 또는 50 mM 내지 150 mM의 농도로 포함하는 약제학적 조성물.The pharmaceutical composition according to any one of claims 20 to 36, comprising an amino acid other than histidine in a concentration of 5 mM to 30 mM or 50 mM to 150 mM.
  38. 제20항 내지 제37항 중 어느 한 항에 있어서, 상기 항-Her2 항체 : 히스티딘 이외의 아미노산의 몰비는 1:10 내지 1:100, 300 : 1 내지 450:1 또는 500:1 내지 700:1인 약제학적 조성물.The method according to any one of claims 20 to 37, wherein the molar ratio of the anti-Her2 antibody: amino acids other than histidine is 1:10 to 1:100, 300:1 to 450:1, or 500:1 to 700:1 Phosphorus pharmaceutical composition.
  39. 제20항 내지 제38항 중 어느 한 항에 있어서, 상기 항-Her2 항체는 퍼투주맙(pertuzumab)인 것인 약제학적 조성물.39. The pharmaceutical composition of any one of claims 20-38, wherein the anti-Her2 antibody is pertuzumab.
  40. 제20항 내지 제39항 중 어느 한 항에 있어서, 상기 항-Her2 항체를 20 mg/ml 내지 150mg/ml의 농도로 포함하는 약제학적 조성물.40. The pharmaceutical composition according to any one of claims 20 to 39, comprising the anti-Her2 antibody at a concentration of 20 mg/ml to 150 mg/ml.
  41. 제20항 내지 제40항 중 어느 한 항에 있어서, 20 mg/ml 내지 30mg/ml의 농도로 항-Her2 항체를 포함하는 IV 투여용 약제학적 조성물.The pharmaceutical composition for IV administration according to any one of claims 20 to 40, comprising an anti-Her2 antibody at a concentration of 20 mg/ml to 30 mg/ml.
  42. 제20항 내지 제41항 중 어느 한 항에 있어서, 폴리솔베이트 20 또는 폴리솔베이트 80을 추가로 포함하는 약제학적 조성물.42. The pharmaceutical composition according to any one of claims 20 to 41, further comprising polysorbate 20 or polysorbate 80.
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