TW201925231A - Pharmaceutical composition comprising anti-human alpha9 integrin antibody - Google Patents

Abstract

Provided is a stable pharmaceutical composition comprising an anti-human [alpha]9 integrin antibody, capable of inhibiting the formation of multimers, degradants, insoluble foreign matters, or insoluble microparticles. The pharmaceutical composition comprises a pharmaceutically acceptable buffer, and has a pH of 5.0 to 7.0.

Description

含有抗人類α９整合素抗體之醫藥組成物Pharmaceutical composition containing anti-human α9 integrin antibody

本發明係有關含有抗人類α9整合素抗體所構成之穩定的醫藥組成物。The present invention relates to a stable pharmaceutical composition comprising an anti-human α9 integrin antibody.

整合素(integrin)係細胞表面糖蛋白質之一，主要功能為細胞與細胞外間質(膠原蛋白、基板蛋白等)的接著，以及作為免疫球蛋白家族成員(ICAM-1、VCAM-1等)之受體，係與傳達來自細胞外間質的信號有關的接著分子。因此，細胞接收來自細胞外間質的信號後，被誘導分化、增殖、細胞凋亡等。整合素係由α鏈與β鏈此二亞單位所構成的異二聚體，存在相異的α鏈、β鏈而有各式各樣的組合，整合素超家族存在24種種類。由於整合素剔除基因小鼠全數因亞單位的變異造成致死或疾病，暗示了整合素係維持生命所必須的。因而促使細胞對應周圍環境傳來訊息的整合素，在生命現象的各種狀況下發揮功能，認為與各式各樣病態有關。Integrin is one of the cell surface glycoproteins. Its main function is the attachment of cells to the extracellular matrix (collagen, substrate protein, etc.) and as members of the immunoglobulin family (ICAM-1, VCAM-1, etc.). The receptor is a follower molecule that signals signals from the extracellular matrix. Therefore, when the cells receive signals from the extracellular matrix, they are induced to differentiate, proliferate, and apoptotic. Integrin is a heterodimer composed of two subunits of α chain and β chain, and there are various combinations of α chain and β chain, and there are 24 types of integrin superfamily. Because integrin knockout mice are all caused by death or disease due to mutations in subunits, suggesting that integrins are essential for life. Therefore, the integrins that cause the cells to transmit information to the surrounding environment function in various conditions of life phenomena, and are thought to be related to various morbid states.

本申請團隊目前已發表抗人類α9整合素抗體係可適用於與病態形成有關的各種疾病之診斷、預防或治療(例如類風濕性關節炎等自體免疫疾病、過敏與移植物排斥等免疫疾病等)(專利文件1)。The application team has published an anti-human α9 integrin anti-system that can be applied to the diagnosis, prevention or treatment of various diseases related to pathogenesis (such as autoimmune diseases such as rheumatoid arthritis, allergic and graft rejection, etc.) Etc.) (Patent Document 1).

另一方面，近年來開發有各式各樣含有抗體等蛋白質的醫藥品，並實際地提供於臨床醫療。由於該等醫藥品大多藉由靜脈內投予與皮下投予等進行投藥，以液體製劑或冷凍乾燥製劑等非經口醫藥組成物之型態提供予臨床醫療。其中特別非經口醫藥組成物由於以直接投予至體內為前提，因而期望穩定的醫藥品製劑。On the other hand, in recent years, various pharmaceutical products containing proteins such as antibodies have been developed and actually provided in clinical medicine. Since many of these pharmaceuticals are administered by intravenous administration, subcutaneous administration, etc., they are provided to clinical medicine in the form of a non-oral pharmaceutical composition such as a liquid preparation or a freeze-dried preparation. Among them, a particularly non-oral pharmaceutical composition is premised on direct administration to the body, and thus a stable pharmaceutical preparation is desired.

含有抗體等蛋白質的溶液中，期望抑制不溶性微粒子的生成等，另外，自有效性的觀點，期望抑制分解物等之生成。In a solution containing a protein such as an antibody, it is desirable to suppress the formation of insoluble fine particles, and it is desirable to suppress the formation of a decomposition product or the like from the viewpoint of effectiveness.

專利文件1中，揭示了用於本發明之抗人類α9整合素抗體本身係對熱穩定。然而，並無與提供於臨床醫療的穩定的醫藥組成物有關之揭示。另外，專利文件2中，揭示含有抗IgE抗體E25等之醫藥組成物。然而，並無與含有抗人類α9整合素抗體所構成，且提供於臨床醫療之穩定的醫藥組成物之揭示。
[先前技術文件]
[專利文件]Patent Document 1 discloses that the anti-human α9 integrin antibody used in the present invention is itself thermostable. However, there is no disclosure relating to stable pharmaceutical compositions provided for clinical care. Further, Patent Document 2 discloses a pharmaceutical composition containing an anti-IgE antibody E25 or the like. However, there is no disclosure of a pharmaceutical composition comprising an anti-human α9 integrin antibody and providing stability in clinical medicine.
[Previous Technical Document]
[Patent Document]

[專利文件1] 國際公開第2009/088064號公報
[專利文件2] 國際公開第2004/091658號公報[Patent Document 1] International Publication No. 2009/088064
[Patent Document 2] International Publication No. 2004/091658

[發明欲解決之課題][Questions to be solved by the invention]

開發含有抗人類α9整合素抗體所構成之更為穩定的醫藥組成物，仍有改善的餘地。本發明的目的係提供含有抗人類α9整合素抗體所構成之穩定的醫藥組成物。There is still room for improvement in the development of a more stable pharmaceutical composition comprising an anti-human α9 integrin antibody. The object of the present invention is to provide a stable pharmaceutical composition comprising an anti-human α9 integrin antibody.

更詳細而言，本發明之目的係提供含有抗人類α9整合素抗體，且可抑制因例如熱、光等壓力而增大之分解物，或是聚合物之生成，所構成之穩定的醫藥組成物。另外，本發明另一目的係提供含有抗人類α9整合素抗體，且可抑制因例如冷凍融解、物理性的震動等而增大之不溶性異物，或是不溶性微粒子的生成，所構成之穩定的醫藥組成物。

[解決課題之手段]More specifically, the object of the present invention is to provide a stable pharmaceutical composition comprising an anti-human α9 integrin antibody and inhibiting decomposition products which are increased by pressure such as heat or light, or formation of a polymer. Things. Further, another object of the present invention is to provide a stable medicine comprising an anti-human α9 integrin antibody and inhibiting the formation of insoluble foreign matter which is increased by, for example, freezing or thawing, physical vibration, or insoluble fine particles. Composition.

[Means for solving the problem]

本發明團隊藉由將含有抗人類α9整合素抗體溶液的pH，調整至特定範圍後進行製劑化，可調製穩定的醫藥組成物(後述實施例1等)，另外，發現藉由添加特定的緩衝劑(後述實施例2等)，添加特定的胺基酸、鹽類、糖或糖醇類(後述實施例3等)，添加非離子性界面活性劑(後述實施例4等)等，可提供更為穩定的醫藥組成物等，本發明遂至完成。The present invention can prepare a stable pharmaceutical composition (Example 1 and the like described later) by adjusting the pH of the anti-human α9 integrin antibody solution to a specific range, and further, it is found that by adding a specific buffer Addition of a specific amino acid, a salt, a sugar, or a sugar alcohol (Example 3, etc. mentioned later), and a nonionic surfactant (The Example 4 etc. mentioned later) etc. are provided, and can provide it. A more stable pharmaceutical composition, etc., is completed by the present invention.

亦即，本發明係關於下述內容。
[1]一種醫藥組成物，其係含有抗人類α9整合素抗體、製藥學容許的緩衝劑，且pH為5.0～7.0之醫藥組成物，
該抗人類α9整合素抗體係選自由下述(1)及(2)所成之群：
(1)含有以序列編號1所示胺基酸序列構成之重鏈及以序列編號3所示胺基酸序列構成之輕鏈之抗人類α9整合素抗體，以及，
(2)藉由(1)之抗人類α9整合素抗體的轉譯後修飾產生之抗體。
[2]如[1]之醫藥組成物，其中製藥學容許的緩衝劑係選自由乙酸、檸檬酸、磷酸、組胺酸，以及該等於製藥學容許的鹽所成群之1種或2種以上。
[3]如[1]或[2]之醫藥組成物，其中製藥學容許的緩衝劑係選自組胺酸或其製藥學容許的鹽。
[4]如[1]～[3]任一項之醫藥組成物，其中製藥學容許的緩衝劑之濃度係1mmol/L～300mmol/L。
[5]如[1]～[4]任一項之醫藥組成物，其係進而含有選自由胺基酸類、鹽類、糖及糖醇類所成群之1種或2種以上。
[6]如[5]之醫藥組成物，其中胺基酸類、鹽類、糖或糖醇類係選自由甲硫胺酸、精胺酸、甘胺酸、離胺酸、鳥胺酸以及該等於製藥學容許的鹽、氯化鈉、蔗糖、山梨糖醇、甘露醇、海藻糖、木糖醇、赤藻糖醇、蘇糖醇、肌醇、半乳糖醇、***糖醇、異麥芽酮糖醇、乳糖醇及麥芽糖醇所成群之1種或2種以上。
[7]如[5]或[6]之醫藥組成物，其中胺基酸類、鹽類、糖或糖醇類係精胺酸。
[8]如[5]～[7]任一項之醫藥組成物，其中胺基酸類、鹽類、糖或糖醇類之濃度係1mmol/L～400mmol/L。
[9]如[1]～[8]任一項之醫藥組成物，其係進而含有非離子性界面活性劑。
[10]如[9]之醫藥組成物，其中非離子性界面活性劑係聚山梨醇酯80及/或波洛莎姆188。
[11]如[9]或[10]之醫藥組成物，其中非離子性界面活性劑之濃度係0.001％(w/v)～1％(w/v)。
[12]如[1]～[11]任一項之醫藥組成物，其中抗人類α9整合素抗體之濃度係1mg/mL～1000mg/mL。
[13]如[1]～[12]任一項之醫藥組成物，其中醫藥組成物係液體製劑或冷凍乾燥製劑。
[14]如[1]～[13]任一項之醫藥組成物，其中醫藥組成物係液體製劑。
[15]一種醫藥組成物，其係含有抗人類α9整合素抗體、組胺酸或其製藥學容許的鹽，且pH為5.0～7.0之醫藥組成物，
該抗人類α9整合素抗體係選自由下述(1)及(2)所成之群：
(1)含有以序列編號1所示胺基酸序列構成之重鏈及以序列編號3所示胺基酸序列構成之輕鏈之抗人類α9整合素抗體，以及，
(2)藉由(1)之抗人類α9整合素抗體的轉譯後修飾產生之抗體。
[16]如[1]或[15]之醫藥組成物，其中抗人類α9整合素抗體的轉譯後修飾係重鏈可變區域N端的焦麩胺醯化以及重鏈C端的離胺酸缺失。
[17]一種使用，其係用以製造含有抗人類α9整合素抗體之穩定的醫藥組成物之製藥學容許的緩衝劑、胺基酸類、鹽類、糖或糖醇類的作為光穩定化劑之使用，
該抗人類α9整合素抗體係選自由下述(1)及(2)所成之群：
(1)含有以序列編號1所示胺基酸序列構成之重鏈及以序列編號3所示胺基酸序列構成之輕鏈之抗人類α9整合素抗體，以及，
(2)藉由(1)之抗人類α9整合素抗體的轉譯後修飾產生之抗體。
[18]如[17]之使用，其中製藥學容許的緩衝劑係選自由乙酸、檸檬酸、磷酸、組胺酸，以及該等於製藥學容許的鹽所成群之1種或2種以上。
[19]一種方法，其係於含有抗人類α9整合素抗體之醫藥組成物中，利用製藥學容許的緩衝劑、胺基酸類、鹽類、糖或糖醇類，抑制抗人類α9整合素抗體因曝光而生成分解物或聚合物之方法，
該抗人類α9整合素抗體係選自由下述(1)及(2)所成之群：
(1)含有以序列編號1所示胺基酸序列構成之重鏈及以序列編號3所示胺基酸序列構成之輕鏈之抗人類α9整合素抗體，以及，
(2)藉由(1)之抗人類α9整合素抗體的轉譯後修飾產生之抗體。
[20]如[19]之方法，其中製藥學容許的緩衝劑係選自由乙酸、檸檬酸、磷酸、組胺酸，以及該等於製藥學容許的鹽所成群之1種或2種以上

[發明的效果]That is, the present invention relates to the following.
[1] A pharmaceutical composition comprising a pharmaceutical composition comprising an anti-human α9 integrin antibody, a pharmaceutically acceptable buffer, and having a pH of 5.0 to 7.0,
The anti-human α9 integrin anti-system is selected from the group consisting of the following (1) and (2):
(1) an anti-human α9 integrin antibody comprising a heavy chain consisting of the amino acid sequence of SEQ ID NO: 1 and a light chain consisting of the amino acid sequence of SEQ ID NO: 3, and
(2) An antibody produced by post-translational modification of the anti-human α9 integrin antibody of (1).
[2] The pharmaceutical composition according to [1], wherein the pharmaceutically acceptable buffer is selected from the group consisting of acetic acid, citric acid, phosphoric acid, histidine, and one or two of the pharmaceutically acceptable salts. the above.
[3] The pharmaceutical composition according to [1] or [2], wherein the pharmaceutically acceptable buffer is selected from the group consisting of histidine or a pharmaceutically acceptable salt thereof.
[4] The pharmaceutical composition according to any one of [1] to [3] wherein the concentration of the pharmaceutically acceptable buffer is from 1 mmol/L to 300 mmol/L.
[5] The pharmaceutical composition according to any one of [1] to [4], which further comprises one or more selected from the group consisting of amino acids, salts, sugars, and sugar alcohols.
[6] The pharmaceutical composition according to [5], wherein the amino acid, the salt, the sugar or the sugar alcohol is selected from the group consisting of methionine, arginine, glycine, lysine, and auramine; Equal to the pharmaceutically acceptable salt, sodium chloride, sucrose, sorbitol, mannitol, trehalose, xylitol, erythritol, threitol, inositol, galactitol, arabitol, isomalt One or two or more kinds of ketitol, lactitol, and maltitol are grouped.
[7] The pharmaceutical composition according to [5] or [6] wherein the amino acid, the salt, the sugar or the sugar alcohol is arginine.
[8] The pharmaceutical composition according to any one of [5] to [7] wherein the concentration of the amino acid, the salt, the sugar or the sugar alcohol is from 1 mmol/L to 400 mmol/L.
[9] The pharmaceutical composition according to any one of [1] to [8] further comprising a nonionic surfactant.
[10] The pharmaceutical composition according to [9], wherein the nonionic surfactant is polysorbate 80 and/or Poloxamm 188.
[11] The pharmaceutical composition according to [9] or [10] wherein the concentration of the nonionic surfactant is 0.001% (w/v) to 1% (w/v).
[12] The pharmaceutical composition according to any one of [1] to [11] wherein the concentration of the anti-human α9 integrin antibody is from 1 mg/mL to 1000 mg/mL.
[13] The pharmaceutical composition according to any one of [1] to [12] wherein the pharmaceutical composition is a liquid preparation or a freeze-dried preparation.
[14] The pharmaceutical composition according to any one of [1] to [13] wherein the pharmaceutical composition is a liquid preparation.
[15] A pharmaceutical composition comprising a pharmaceutical composition comprising an anti-human α9 integrin antibody, histidine or a pharmaceutically acceptable salt thereof, and having a pH of 5.0 to 7.0,
The anti-human α9 integrin anti-system is selected from the group consisting of the following (1) and (2):
(1) an anti-human α9 integrin antibody comprising a heavy chain consisting of the amino acid sequence of SEQ ID NO: 1 and a light chain consisting of the amino acid sequence of SEQ ID NO: 3, and
(2) An antibody produced by post-translational modification of the anti-human α9 integrin antibody of (1).
[16] The pharmaceutical composition according to [1] or [15], wherein the post-translational modification of the anti-human α9 integrin antibody is a glutamate deuteration at the N-terminus of the heavy chain variable region and an acyl acid depletion at the C-terminus of the heavy chain.
[17] A method for producing a pharmaceutically acceptable buffer, an amino acid, a salt, a sugar or a sugar alcohol as a photostabilizer for a stable pharmaceutical composition comprising an anti-human α9 integrin antibody Use,
The anti-human α9 integrin anti-system is selected from the group consisting of the following (1) and (2):
(1) an anti-human α9 integrin antibody comprising a heavy chain consisting of the amino acid sequence of SEQ ID NO: 1 and a light chain consisting of the amino acid sequence of SEQ ID NO: 3, and
(2) An antibody produced by post-translational modification of the anti-human α9 integrin antibody of (1).
[18] The use of the pharmaceutically acceptable buffer is one or more selected from the group consisting of acetic acid, citric acid, phosphoric acid, histidine, and the pharmaceutically acceptable salt.
[19] A method for inhibiting an anti-human α9 integrin antibody by using a pharmaceutically acceptable buffer, an amino acid, a salt, a sugar or a sugar alcohol in a pharmaceutical composition containing an anti-human α9 integrin antibody a method of generating a decomposition product or a polymer by exposure,
The anti-human α9 integrin anti-system is selected from the group consisting of the following (1) and (2):
(1) an anti-human α9 integrin antibody comprising a heavy chain consisting of the amino acid sequence of SEQ ID NO: 1 and a light chain consisting of the amino acid sequence of SEQ ID NO: 3, and
(2) An antibody produced by post-translational modification of the anti-human α9 integrin antibody of (1).
[20] The method according to [19], wherein the pharmaceutically acceptable buffer is selected from the group consisting of acetic acid, citric acid, phosphoric acid, histidine, and one or more selected from the group consisting of pharmaceutically acceptable salts.

[Effects of the Invention]

根據本發明，可提供含有抗人類α9整合素抗體所構成之穩定的醫藥組成物，詳細係可提供抑制聚合物、分解物、不溶性異物或不溶性微粒子的生成所構成之穩定的醫藥組成物。According to the present invention, it is possible to provide a stable pharmaceutical composition comprising an anti-human α9 integrin antibody, and in detail, it is possible to provide a stable pharmaceutical composition comprising a polymer, a decomposition product, an insoluble foreign matter or insoluble microparticles.

本說明書中「穩定」係指例如對熱、光、溫度、濕度、振盪及/或冷凍融解為穩定之意。例如，於規定條件下保存醫藥組成物後，將前述醫藥組成物中含有的聚合物與分解物的總量，或者該等的增加量規定於特定量以下。一實施方式係將電荷類似物規定於特定量以下。另外，一實施方式係將於規定條件下保存醫藥組成物後，以目視未觀察到不溶性異物，或者，利用後述光遮斷粒子計數法測定不溶性微粒子，並規定於特定數以下。"Stable" in this specification means, for example, that heat, light, temperature, humidity, oscillation, and/or freeze-thaw is stable. For example, after the pharmaceutical composition is stored under predetermined conditions, the total amount of the polymer and the decomposed product contained in the pharmaceutical composition or the amount of the increase is set to a specific amount or less. In one embodiment, the charge analog is specified below a certain amount. In addition, in one embodiment, after the pharmaceutical composition is stored under predetermined conditions, insoluble foreign matter is not observed by visual observation, or the insoluble fine particles are measured by a light-blocking particle counting method described later, and are specified to be a specific number or less.

前述「聚合物」係抗人類α9整合素抗體聚集後生成的複合體。並未特別限制測定聚合物的方法，例如包含粒徑篩析層析法(SE-HPLC法)、凝膠電泳法、毛細管電泳法、動態光散亂法、光遮斷粒子計數法、流式顯微影像顆粒分析法等，一實施方法係SE-HPLC法。SE-HPLC法中可利用自動分析法測定檢出的波峰面積，再藉由除以包含主要波峰的全波峰面積的總和，計算出百分率(％)作為各聚合物之量。且主要波峰係意指活性本體的波峰。合併較SE-HPLC的主要波峰維持時間為短的波峰，作為聚合物的總量(以下稱作聚合物量)。聚合物量，一實施方式係10％以下，一實施方式係5％以下，一實施方式係3％以下。另外，針對聚合物量的增加量(於特定條件下、特定期間的保存後的聚合物量，與開始試驗(保存)時聚合物量之差)，一實施方式係5％以下，一實施方式係2％以下，一實施方式係1％以下，一實施方式係0.5％以下。且，針對前述聚合物量或聚合物量之增加量，可依據後述「穩定的醫藥組成物」定義所記載的各保存條件，與依據期望任意地組合使用。The aforementioned "polymer" is a complex produced by aggregation of an anti-human α9 integrin antibody. The method for measuring the polymer is not particularly limited, and includes, for example, particle size exclusion chromatography (SE-HPLC method), gel electrophoresis, capillary electrophoresis, dynamic light scattering, photo-interrupting particle counting, flow Microscopic image particle analysis method, etc., an implementation method is SE-HPLC method. In the SE-HPLC method, the detected peak area can be measured by an automatic analysis method, and the percentage (%) is calculated as the amount of each polymer by dividing by the sum of the full peak areas including the main peaks. And the main peak system means the peak of the active body. The main peak maintenance time of the combined SE-HPLC was a short peak as the total amount of the polymer (hereinafter referred to as the amount of the polymer). The amount of the polymer is 10% or less in one embodiment, 5% or less in one embodiment, and 3% or less in one embodiment. Further, the amount of increase in the amount of the polymer (the difference between the amount of the polymer after storage under a specific condition and a specific period and the amount of the polymer at the time of starting the test (storage)) is 5% or less in one embodiment, and 2% in one embodiment. Hereinafter, one embodiment is 1% or less, and one embodiment is 0.5% or less. In addition, the amount of increase in the amount of the polymer or the amount of the polymer can be used arbitrarily in combination with the desired storage conditions according to the definition of "stable pharmaceutical composition" described later.

前述之「分解物」係抗人類α9整合素抗體的一部分脫離後生成的斷片體。並未特別限制測定分解物的方法，例如包含SE-HPLC法、凝膠電泳法、毛細管電泳法等，一實施方法係SE-HPLC法。利用SE-HPLC法之分解物的測定方法，可利用與前述SE-HPLC法之聚合物測定法相同方式進行。且，合併較SE-HPLC的主要波峰維持時間為長的波峰，作為分解物的總量(以下稱作分解物量)。分解物量，一實施方式係10％以下，一實施方式係5％以下，一實施方式係3％以下。另外，針對分解物量的增加量(於特定條件下、特定期間的保存後的分解物量，與開始試驗(保存)時分解物量之差)，一實施方式係10％以下，一實施方式係5％以下，一實施方式係3％以下。且，針對前述分解物量或分解物量之增加量，可依據後述「穩定的醫藥組成物」定義所記載的各保存條件，與依據期望任意地組合使用。The "decomposed product" described above is a fragment formed by detachment of a part of the human α9 integrin antibody. The method for measuring the decomposition product is not particularly limited, and includes, for example, SE-HPLC method, gel electrophoresis method, capillary electrophoresis method, etc., and one embodiment is SE-HPLC method. The method for measuring the decomposition product by the SE-HPLC method can be carried out in the same manner as the polymer measurement method of the SE-HPLC method described above. Further, the main peak holding time of the SE-HPLC was combined as a long peak as the total amount of the decomposition product (hereinafter referred to as the amount of the decomposition product). The amount of the decomposition product is 10% or less in one embodiment, 5% or less in one embodiment, and 3% or less in one embodiment. In addition, the amount of the amount of the decomposition product (the difference between the amount of the decomposition product after storage under a specific condition and the specific period and the amount of the decomposition product at the time of starting the test (storage)) is 10% or less in one embodiment, and 5% in one embodiment. Hereinafter, one embodiment is 3% or less. In addition, the amount of the decomposition product or the amount of the decomposition product can be used arbitrarily in combination with the desired storage conditions according to the definition of "stable pharmaceutical composition" described later.

前述「電荷類似物」係利用陽離子交換管柱層析法，或毛細管等電聚焦電泳法分析，在主要波峰之外可發現的類似物。並未特別限制測定電荷類似物的方法，例如包含陽離子交換管柱層析法(CE-HPLC法)、毛細管等電聚焦電泳法(icIEF法)等。一實施方法係CE-HPLC法，或icIEF法。CE-HPLC法及icIEF法可利用與前述SE-HPLC法同樣地進行，計算出波峰的百分率(％)，再作為各電荷類似物的量。電荷類似物的總量，一實施方式係0～70％，一實施方式係0～50％，一實施方式係0～30％，一實施方式係0～10％。The aforementioned "charge analog" is an analog which can be found outside the main peak by cation exchange column chromatography or capillary isoelectric focusing electrophoresis. The method for measuring the charge analog is not particularly limited, and includes, for example, cation exchange column chromatography (CE-HPLC method), capillary isoelectric focusing electrophoresis (ICIEF method), and the like. One method of implementation is the CE-HPLC method, or the icIEF method. The CE-HPLC method and the icIEF method can be carried out in the same manner as the above-described SE-HPLC method, and the percentage (%) of the peaks is calculated and used as the amount of each charge analog. The total amount of charge analogs is 0 to 70% in one embodiment, 0 to 50% in one embodiment, 0 to 30% in one embodiment, and 0 to 10% in one embodiment.

前述「不溶性異物」係可遵循日本藥典有關注射劑的不溶性異物檢查法，可藉由目視確認。The aforementioned "insoluble foreign matter" can be inspected by the Japanese Pharmacopoeia in accordance with the insoluble foreign matter inspection method for injections, and can be visually confirmed.

前述「不溶性微粒子」係抗人類α9整合素抗體與醫藥添加物等聚集而生成的不溶性的複合體。不溶性微粒子的數量可藉由光遮斷粒子計數法，並可規定不溶性微粒子的數量為特定數以下。一實施方式，1.2μm以上的不溶性微粒子的數量為1000個/mL以下。The "insoluble fine particles" are insoluble complexes formed by aggregation of anti-human α9 integrin antibodies and pharmaceutical additives. The number of insoluble fine particles can be determined by photo-interrupting particle counting, and the number of insoluble fine particles can be specified to be a specific number or less. In one embodiment, the number of insoluble fine particles of 1.2 μm or more is 1000/mL or less.

「穩定的醫藥組成物」係意指於冷藏溫度(2℃～8℃)例如3個月期間，一實施方式為6個月期間，一實施方式為1年間，一實施方式為2年間；或室溫(22℃～28℃)例如2週間，一實施方式為1個月期間，一實施方式為3個月期間，一實施方式為6個月期間，一實施方式為1年間；或加速條件(37℃～43℃)例如2週間，一實施方式為4週期間；或於D65燈1000lux照射下，例如48小時，一實施方式為168小時曝光後，聚合物量或分解物量的增加量，或者聚合物量或分解物量為特定量以下的醫藥組成物。The "stable pharmaceutical composition" means a refrigerating temperature (2 ° C to 8 ° C), for example, a three-month period, and an embodiment is a six-month period, one embodiment is one year, and one embodiment is two years; or Room temperature (22 ° C ~ 28 ° C), for example, 2 weeks, one embodiment is a one-month period, one embodiment is a three-month period, one embodiment is a six-month period, and one embodiment is one year; or an acceleration condition (37 ° C ~ 43 ° C), for example, 2 weeks, an embodiment of 4 weeks; or D65 lamp 1000 lux irradiation, for example 48 hours, an embodiment of 168 hours of exposure, the amount of polymer or decomposition of the amount of increase, or The amount of the polymer or the amount of the decomposition product is a specific amount or less of the pharmaceutical composition.

本發明之醫藥組成物中所含抗人類α9整合素抗體(亦稱為「可用於本發明之抗人類α9整合素抗體」)，係含有下述(1)及/或(2)之抗人類α9整合素抗體：
(1)包含序列編號1表示之胺基酸序列構成之重鏈以及序列編號3表示之胺基酸序列構成之輕鏈之抗人類α9整合素抗體(國際公開第2009/088064號公報)
(2)(1)之抗人類α9整合素抗體轉譯後修飾產生之抗體。The anti-human α9 integrin antibody (also referred to as "anti-human α9 integrin antibody useful in the present invention" contained in the pharmaceutical composition of the present invention contains the following antibiotics (1) and/or (2). 99 integrin antibody:
(1) An anti-human α9 integrin antibody comprising a heavy chain composed of an amino acid sequence represented by SEQ ID NO: 1 and a light chain composed of an amino acid sequence represented by SEQ ID NO: 3 (International Publication No. 2009/088064)
(2) An antibody produced by post-translational modification of the anti-human α9 integrin antibody of (1).

使細胞表現抗體時，已知抗體係於轉譯後受到修飾。轉譯後修飾之例可舉出重鏈C端的離胺酸因羧肽酶而切斷、重鏈及輕鏈N端的麩醯胺酸或麩胺酸因焦麩胺酸化而修飾為焦麩胺酸等，已知於各種抗體，重鏈C端的離胺酸缺失與重鏈N端的麩醯胺酸的大部分受到修飾成為焦麩胺酸(Journal of Pharmaceutical Sciences, 2008,Vol. 97, p. 2426-2447)。另外，該等轉譯後修飾不會對抗體的活性造成影響亦係該領域內熟知者(Analytical Biochemistry, 2006, Vol. 348, p. 24-39)。When the cells are expressed as antibodies, the anti-system is known to be modified after translation. Examples of the post-translational modification include the cleavage of the amino acid at the C-terminus of the heavy chain by carboxypeptidase, and the glutamic acid or glutamic acid at the N-terminus of the heavy chain and the light chain are modified to be glutamic acid by acidification of the glutamate. Etc., known in various antibodies, the cleavage of the leucine at the C-terminus of the heavy chain and the majority of the glutamic acid at the N-terminus of the heavy chain have been modified to be glutamate (Journal of Pharmaceutical Sciences, 2008, Vol. 97, p. 2426 -2447). In addition, such post-translational modifications do not affect the activity of the antibody and are well known in the art (Analytical Biochemistry, 2006, Vol. 348, p. 24-39).

其中一實施方式中，前述(2)之抗人類α9整合素抗體的轉譯後修飾係重鏈N端的焦麩胺酸化。其中一實施方式中，前述(2)之抗人類α9整合素抗體係包含由胺基酸編號1的麩醯胺酸經修飾為焦麩胺酸之序列編號1的胺基酸序列所構成的重鏈，及由序列編號3所示胺基酸序列構成的輕鏈之抗人類α9整合素抗體。In one embodiment, the post-translational modification of the anti-human α9 integrin antibody of (2) is pyroglutamic acidification of the N-terminus of the heavy chain. In one embodiment, the anti-human α9 integrin anti-system of the above (2) comprises a heavy amino acid sequence consisting of the amino acid sequence of the amino acid number 1 of the amino acid number 1 modified to the pyroglutamic acid. A chain, and a light chain anti-human α9 integrin antibody consisting of the amino acid sequence of SEQ ID NO: 3.

其中一實施方式中，前述(2)之抗人類α9整合素抗體的轉譯後修飾係重鏈C端的離胺酸缺失。其中一實施方式中，前述(2)之抗人類α9整合素抗體係包含由胺基酸編號1的胺基酸編號1至449之胺基酸序列所構成的重鏈，及由序列編號3所示胺基酸序列構成的輕鏈之抗人類α9整合素抗體。In one embodiment, the post-translational modification of the anti-human α9 integrin antibody of (2) above is a deletion of the amino acid at the C-terminus of the heavy chain. In one embodiment, the anti-human α9 integrin anti-system of (2) above comprises a heavy chain composed of an amino acid sequence of amino acid numbers 1 to 449 of amino acid number 1, and is designated by SEQ ID NO: 3 An anti-human α9 integrin antibody of the light chain composed of an amino acid sequence.

其中一實施方式中，前述(2)之抗人類α9整合素抗體的轉譯後修飾係重鏈N端的焦麩胺酸化，且，重鏈C端的離胺酸缺失。其中一實施方式中，前述(2)之抗人類α9整合素抗體係包含由胺基酸編號1的麩醯胺酸經修飾為焦麩胺酸之序列編號1的胺基酸編號1至449之胺基酸序列所構成的重鏈，及由序列編號3所示胺基酸序列構成的輕鏈之抗人類α9整合素抗體。In one embodiment, the post-translational modification of the anti-human α9 integrin antibody of (2) is pyrophorylated at the N-terminus of the heavy chain, and the acyl acid at the C-terminus of the heavy chain is deleted. In one embodiment, the anti-human α9 integrin anti-system of the above (2) comprises amino acid number 1 to 449 of SEQ ID NO: 1 modified with amino acid 1 of branic acid. A heavy chain composed of an amino acid sequence, and a light chain anti-human α9 integrin antibody consisting of the amino acid sequence of SEQ ID NO: 3.

可用於本發明之人類化抗人類α9整合素抗體，尚可舉出藉由培養下述宿主細胞可生產的抗人類α9整合素抗體：
經以含有包含編碼序列編號1所示胺基酸序列構成的重鏈之鹼基序列之聚核苷酸，以及包含編碼序列編號3所示胺基酸序列構成的輕鏈之鹼基序列之聚核苷酸之表現載體，進行轉形作用(Transformation)的宿主細胞，或者，
經以含有包含編碼序列編號1所示胺基酸序列構成的重鏈之鹼基序列之聚核苷酸之表現載體，以及含有包含編碼序列編號3所示胺基酸序列構成的輕鏈之鹼基序列之聚核苷酸之表現載體，進行轉形作用(Transformation)的宿主細胞。The humanized anti-human α9 integrin antibody which can be used in the present invention is exemplified by an anti-human α9 integrin antibody which can be produced by culturing the following host cells:
Polynucleotide comprising a base sequence comprising a heavy chain comprising the amino acid sequence of coding sequence number 1 and a base sequence comprising a light chain comprising the amino acid sequence of SEQ ID NO: 3 a nucleotide expression vector, a host cell for transformation, or
An expression vector comprising a polynucleotide comprising a base sequence comprising a heavy chain consisting of the amino acid sequence of SEQ ID NO: 1, and a base comprising a light chain comprising the amino acid sequence of SEQ ID NO: The expression vector of the polynucleotide of the base sequence is subjected to a transformation of the host cell.

可用於本發明之抗人類α9整合素抗體係基於本說明書中揭示的抗人類α9整合素抗體的序列資訊為基礎，使用相關領域周知的方法，相關業者可容易地製作。本發明可使用之抗人類α9整合素抗體的製作方法，可舉出國際公開第2009/088064號公報揭示的方法。The anti-human α9 integrin anti-system useful in the present invention is based on the sequence information of the anti-human α9 integrin antibody disclosed in the present specification, and can be easily produced by a related art using a method well known in the related art. The method for producing an anti-human α9 integrin antibody which can be used in the present invention includes the method disclosed in International Publication No. 2009/088064.

一單位醫藥組成物(製劑)中，本發明可使用的抗人類α9整合素抗體之量，可舉出例如1mg～1000mg，一實施方式係1mg～500mg，一實施方式係1mg～30mg，一實施方式係10mg～200mg，一實施方式係100mg～250mg，一實施方式係50mg～250mg，一實施方式係100mg～200mg。且，前述的上限與下限可依據期望任意地加以組合。In one unit of the pharmaceutical composition (formulation), the amount of the anti-human α9 integrin antibody which can be used in the present invention is, for example, 1 mg to 1000 mg, and in one embodiment, 1 mg to 500 mg, and in one embodiment, 1 mg to 30 mg. The method is 10 mg to 200 mg, and in one embodiment, 100 mg to 250 mg, in one embodiment, 50 mg to 250 mg, and in one embodiment, 100 mg to 200 mg. Moreover, the aforementioned upper and lower limits may be arbitrarily combined as desired.

醫藥組成物為固體狀態時(例如冷凍乾燥製劑、噴霧乾燥製劑等)，本發明可使用的抗人類α9整合素抗體之量，可舉出例如1mg～1000mg，一實施方式係1mg～500mg，一實施方式係1mg～30mg，一實施方式係10mg～200mg，一實施方式係100mg～250mg，一實施方式係50mg～250mg，一實施方式係100mg～200mg。且，前述的上限與下限可依據期望任意地加以組合。When the pharmaceutical composition is in a solid state (for example, a freeze-dried preparation, a spray-dried preparation, or the like), the amount of the anti-human α9 integrin antibody which can be used in the present invention is, for example, 1 mg to 1000 mg, and in one embodiment, 1 mg to 500 mg, The embodiment is 1 mg to 30 mg, and in one embodiment, 10 mg to 200 mg, in one embodiment, 100 mg to 250 mg, in one embodiment, 50 mg to 250 mg, and in one embodiment, 100 mg to 200 mg. Moreover, the aforementioned upper and lower limits may be arbitrarily combined as desired.

使用時溶解醫藥組成物時，本發明可使用的抗人類α9整合素抗體之濃度，可舉出例如1mg/mL～1000mg/mL，一實施方式係1mg/mL～500mg/mL，一實施方式係1mg/mL～30mg/mL，一實施方式係10mg/mL～200mg/mL，一實施方式係100mg/mL～250mg/mL，一實施方式係50mg/mL～250mg/mL，一實施方式係100mg/mL～200mg/mL。且，前述的上限與下限可依據期望任意地加以組合。When the pharmaceutical composition is dissolved at the time of use, the concentration of the anti-human α9 integrin antibody which can be used in the present invention is, for example, 1 mg/mL to 1000 mg/mL, and in one embodiment, 1 mg/mL to 500 mg/mL. 1 mg/mL to 30 mg/mL, one embodiment is 10 mg/mL to 200 mg/mL, one embodiment is 100 mg/mL to 250 mg/mL, and one embodiment is 50 mg/mL to 250 mg/mL, and one embodiment is 100 mg/mL. mL ~ 200mg / mL. Moreover, the aforementioned upper and lower limits may be arbitrarily combined as desired.

醫藥組成物為溶液狀態(液體製劑)時，本發明可使用的抗人類α9整合素抗體之濃度，可舉出例如1mg/mL～1000mg/mL，一實施方式係1mg/mL～500mg/mL，一實施方式係1mg/mL～30mg/mL，一實施方式係10mg/mL～200mg/mL，一實施方式係100mg/mL～250mg/mL，一實施方式係50mg/mL～250mg/mL，一實施方式係100mg/mL～200mg/mL。且，前述的上限與下限可依據期望任意地加以組合。When the pharmaceutical composition is in a solution state (liquid preparation), the concentration of the anti-human α9 integrin antibody which can be used in the present invention is, for example, 1 mg/mL to 1000 mg/mL, and in one embodiment, 1 mg/mL to 500 mg/mL. One embodiment is 1 mg/mL to 30 mg/mL, and one embodiment is 10 mg/mL to 200 mg/mL, and one embodiment is 100 mg/mL to 250 mg/mL, and one embodiment is 50 mg/mL to 250 mg/mL. The method is 100 mg/mL to 200 mg/mL. Moreover, the aforementioned upper and lower limits may be arbitrarily combined as desired.

用量可舉出例如0.002mg/kg～100mg/kg，一實施方式係0.01mg/kg～50mg/kg，一實施方式係1mg/kg～30mg/kg。且，前述的上限與下限可依據期望任意地加以組合。The dosage is, for example, 0.002 mg/kg to 100 mg/kg, and in one embodiment, 0.01 mg/kg to 50 mg/kg, and in one embodiment, 1 mg/kg to 30 mg/kg. Moreover, the aforementioned upper and lower limits may be arbitrarily combined as desired.

適應症可舉出與整合素相關的各種疾病之預防及/或治療，例如類風濕性關節炎等自體免疫疾病、過敏與移植物排斥等免疫疾病等。The indications include prevention and/or treatment of various diseases related to integrin, such as autoimmune diseases such as rheumatoid arthritis, immune diseases such as allergy and graft rejection, and the like.

可使用於本發明之「製藥學上容許的緩衝劑」，若為製藥學上容許，於溶液狀態在期望的pH範圍內具有緩衝能力，或具有穩定化效果者，則無特別限制。
具體而言，於後述pH範圍內具有緩衝能力者。例如，5.0～7.0，一實施方式5.5～6.5，一實施方式5.6～6.2，一實施方式6.0，一實施方式5.8。且，前述的上限與下限可依據期望任意地加以組合。
且本說明書中醫藥組成物的pH係指液體製劑時於溶液狀態時的pH，非液體製劑(例如冷凍乾燥製劑、噴霧乾燥製劑等)時，係意指溶解於溶媒(例如注射用水)的溶液狀態時的pH。The "pharmaceutically acceptable buffer" to be used in the present invention is not particularly limited as long as it is pharmaceutically acceptable, has a buffering ability in a desired pH range in a solution state, or has a stabilizing effect.
Specifically, it has a buffering ability in the pH range mentioned later. For example, 5.0 to 7.0, one embodiment 5.5 to 6.5, one embodiment 5.6 to 6.2, one embodiment 6.0, and one embodiment 5.8. Moreover, the aforementioned upper and lower limits may be arbitrarily combined as desired.
Further, the pH of the pharmaceutical composition in the present specification means the pH in a liquid state in a solution state, and the non-liquid preparation (for example, a freeze-dried preparation, a spray-dried preparation, etc.) means a solution dissolved in a solvent (for example, water for injection). The pH at the time of the state.

緩衝劑的成分，可舉出例如乙酸、檸檬酸、磷酸、組胺酸或該等之製藥學上容許的鹽等。一實施方式係組胺酸或其製藥學上容許的鹽等，一實施方式係離組胺酸。
乙酸、檸檬酸、磷酸、組胺酸及該等之製藥學上容許的鹽不僅可作為緩衝劑成分，亦顯示不會影響本發明可用之抗人類α9整合素抗體的穩定性，或提高穩定性之效果。
另外，乙酸、檸檬酸、磷酸、組胺酸及該等之製藥學上容許的鹽，顯示特別可抑制因曝光之本發明可用之抗人類α9整合素抗體的分解物或聚合物的生成之光穩定化效果。The component of the buffering agent may, for example, be acetic acid, citric acid, phosphoric acid, histidine or a pharmaceutically acceptable salt thereof. One embodiment is a histidine or a pharmaceutically acceptable salt thereof, and one embodiment is a histidine acid.
Acetic acid, citric acid, phosphoric acid, histidine, and the pharmaceutically acceptable salts thereof are not only useful as buffer components, but also show that the stability of the anti-human α9 integrin antibody useful in the present invention is not affected, or stability is improved. The effect.
Further, acetic acid, citric acid, phosphoric acid, histidine, and the pharmaceutically acceptable salts thereof exhibit light which specifically inhibits the decomposition of the anti-human α9 integrin antibody or the polymer produced by the present invention which is exposed. Stabilization effect.

緩衝劑成分可適宜適量使用1種或2種以上。
緩衝劑的濃度若為可將pH調整在期待的pH範圍內之量，則無特別限制。緩衝劑的濃度，於利用溶媒(例如注射用水)溶解的溶液狀態(液體製劑)時，可舉出例如1mmol/L～300mmol/L，一實施方式為，10mmol/L～150mmol/L，一實施方式為，1mmol/L～50mmol/L，一實施方式為，5mmol/L～175mmol/L。且，前述的上限與下限可依據期望任意地加以組合。The buffer component may be used singly or in an appropriate amount in an appropriate amount.
The concentration of the buffer is not particularly limited as long as it can adjust the pH within a desired pH range. When the concentration of the buffer is in a solution state (liquid preparation) dissolved in a solvent (for example, water for injection), for example, 1 mmol/L to 300 mmol/L, and in one embodiment, 10 mmol/L to 150 mmol/L, one implementation is carried out. The method is 1 mmol/L to 50 mmol/L, and in one embodiment, 5 mmol/L to 175 mmol/L. Moreover, the aforementioned upper and lower limits may be arbitrarily combined as desired.

可用於本發明之胺基酸類、鹽類、糖或糖醇類，若為製藥學上容許，且不會影響本發明可用之抗人類α9整合素抗體的穩定性，或可提高穩定性者，則無特別限制。The amino acids, salts, sugars or sugar alcohols which can be used in the present invention, if pharmaceutically acceptable, do not affect the stability of the anti-human α9 integrin antibody useful in the present invention, or can improve stability, There is no special restriction.

胺基酸類可舉出例如甲硫胺酸、精胺酸、甘胺酸、離胺酸、鳥胺酸或該等於製藥學容許的鹽。一實施方式為甲硫胺酸、精胺酸、甘胺酸，一實施方式為精胺酸。
鹽類可舉出例如氯化鈉等。
糖或糖醇類可舉出例如蔗糖、山梨糖醇、甘露醇、海藻糖、木糖醇、赤藻糖醇、蘇糖醇、肌醇、半乳糖醇、***糖醇、異麥芽酮糖醇、乳糖醇及麥芽糖醇等。一實施方式為蔗糖、山梨糖醇，一實施方式為山梨糖醇。
胺基酸類、鹽類、糖或糖醇類，一實施方式為精胺酸、甲硫胺酸、或該等於製藥學容許的鹽，山梨糖醇、蔗糖、氯化鈉，一實施方式為精胺酸。The amino acid may, for example, be methionine, arginine, glycine, lysine, ornithamine or a salt which is equivalent to a pharmaceutically acceptable salt. One embodiment is methionine, arginine, glycine, and one embodiment is arginine.
The salt may, for example, be sodium chloride or the like.
Examples of the sugar or sugar alcohol include sucrose, sorbitol, mannitol, trehalose, xylitol, erythritol, threitol, inositol, galactitol, arabitol, isomaltulose. Alcohol, lactitol and maltitol. One embodiment is sucrose, sorbitol, and one embodiment is sorbitol.
An amino acid, a salt, a sugar or a sugar alcohol, in one embodiment is arginine, methionine, or a pharmaceutically acceptable salt, sorbitol, sucrose, sodium chloride, an embodiment is fine Amino acid.

胺基酸類、鹽類、糖或糖醇類，可適宜使用1種或2種以上。
且，胺基酸類、鹽類、糖或糖醇類的種類及組合，若為不會影響本發明可用之抗人類α9整合素抗體的穩定性，或可提高穩定性者，則無特別限制。
本發明可用之抗人類α9整合素抗體，顯示精胺酸、甲硫胺酸、山梨糖醇、蔗糖、氯化鈉具有對光穩定化之效果。
胺基酸類、鹽類、糖或糖醇類之量，若為不會影響本發明可用之抗人類α9整合素抗體的穩定性，或可提高穩定性者，則無特別限制。
例如，利用溶媒溶解的溶液狀態(液體製劑)時的濃度，1mmol/L～400mmol/L，一實施方式為，1mmol/L～300mmol/L，一實施方式為，100mmol/L～300mmol/L，一實施方式為100mmol/L～200mmol/L。且，前述的上限與下限可依據期望任意地加以組合。The amino acid, the salt, the sugar, or the sugar alcohol may be used singly or in combination of two or more kinds.
Further, the type and combination of amino acids, salts, sugars or sugar alcohols are not particularly limited as long as they do not affect the stability of the anti-human α9 integrin antibody which can be used in the present invention, or can improve the stability.
The anti-human α9 integrin antibody useful in the present invention shows that arginine, methionine, sorbitol, sucrose, and sodium chloride have an effect of stabilizing light.
The amount of the amino acid, the salt, the sugar or the sugar alcohol is not particularly limited as long as it does not affect the stability of the anti-human α9 integrin antibody which can be used in the present invention, or can improve the stability.
For example, the concentration in the solution state (liquid preparation) in which the solvent is dissolved is 1 mmol/L to 400 mmol/L, and in one embodiment, 1 mmol/L to 300 mmol/L, and in one embodiment, 100 mmol/L to 300 mmol/L, One embodiment is from 100 mmol/L to 200 mmol/L. Moreover, the aforementioned upper and lower limits may be arbitrarily combined as desired.

本發明可用之抗人類α9整合素抗體作為光穩定化劑時，可使用製藥學上許可的緩衝劑、胺基酸類、鹽類、糖或糖醇類。例如可舉出乙酸、檸檬酸、磷酸、組胺酸、精胺酸、甲硫胺酸或該等之製藥學上許可的鹽、山梨糖醇、蔗糖、氯化鈉等。When the anti-human α9 integrin antibody which can be used in the present invention is used as a photostabilizer, a pharmaceutically acceptable buffer, an amino acid, a salt, a sugar or a sugar alcohol can be used. For example, acetic acid, citric acid, phosphoric acid, histidine, arginine, methionine or a pharmaceutically acceptable salt thereof, sorbitol, sucrose, sodium chloride or the like can be given.

可用於本發明之非離子性界面活性劑，若為製藥學上許可，且不會影響本發明可用之抗人類α9整合素抗體的穩定化作用者，則無特別限制。The nonionic surfactant which can be used in the present invention is not particularly limited as long as it is pharmaceutically acceptable and does not affect the stabilization of the anti-human α9 integrin antibody which can be used in the present invention.

具體而言，前述非離子性界面活性劑包含例如具有山梨坦單辛酸酯、山梨坦單月桂酸酯，及山梨坦單棕櫚酸酯等山梨坦脂肪酸酯；單辛酸甘油酯、單肉荳蔻酸甘油酯、及單硬酯酸甘油酯等甘油脂肪酸酯；十聚甘油單硬酯酸酯、十聚甘油二硬酯酸酯，及十聚甘油單亞麻油酸酯等聚甘油脂肪酸酯；聚氧乙烯去水山梨醇單月桂酸酯、聚氧乙烯去水山梨醇單油酸酯、聚氧乙烯去水山梨醇單硬酯酸酯、聚氧乙烯去水山梨醇單棕櫚酸酯、聚氧乙烯去水山梨醇三油酸酯，及聚氧乙烯去水山梨醇三硬酯酸酯等聚氧乙烯去水山梨醇脂肪酸酯；聚氧乙烯山梨醇四硬酯酸酯及聚氧乙烯山梨醇四油酸酯等聚氧乙烯山梨醇脂肪酸酯；聚氧乙烯甘油單硬酯酸酯等聚氧乙烯甘油脂肪酸酯；聚乙二醇二硬酯酸酯等聚乙二醇脂肪酸酯；聚氧乙烯月桂醚等聚氧乙烯烷基醚；聚氧乙烯聚氧丙烯二醇醚、聚氧乙烯聚氧丙烯丙基醚，及聚氧乙烯聚氧丙烯鯨蠟基醚等聚氧乙烯聚氧丙烯烷基醚；聚氧乙烯壬基苯基醚等聚氧乙烯烷基苯基醚；聚氧乙烯蓖麻油，及聚氧乙烯硬化蓖麻油(聚氧乙烯氫化蓖麻油)等聚氧乙烯硬化蓖麻油；聚氧乙烯山梨醇蜜蠟等聚氧乙烯蜜蠟衍生物；聚氧乙烯羊毛脂等聚氧乙烯羊毛脂衍生物；聚氧乙烯脂肪醯胺，例如聚氧乙烯十八醯胺等6～18之Hydrophilic-Lipophilic Balance(HLB)之界面活性劑。Specifically, the nonionic surfactant includes, for example, sorbitan fatty acid esters such as sorbitan monocaprylate, sorbitan monolaurate, and sorbitan monopalmitate; glyceryl monocaprylate, single nutmeg Glycerol fatty acid esters such as glycerides and glyceryl monostearate; polyglycerol fatty acid esters such as decaglycerin monostearate, decaglyceryl distearate, and decaglycerin monolinoleic acid ester Polyoxyethylene sorbitan monolaurate, polyoxyethylene sorbitan monooleate, polyoxyethylene sorbitan monostearate, polyoxyethylene sorbitan monopalmitate, Polyoxyethylene sorbitan trioleate, polyoxyethylene sorbitan tristearate, polyoxyethylene sorbitan fatty acid ester, polyoxyethylene sorbitol tetrastearate and polyoxygen Polyoxyethylene sorbitan fatty acid ester such as ethylene sorbitol tetraoleate; polyoxyethylene glycerin fatty acid ester such as polyoxyethylene glycerol monostearate; polyethylene glycol fat such as polyethylene glycol distearate Acid ester; polyoxyethylene alkyl ether such as polyoxyethylene lauryl ether; polyoxyethylene polyoxyl Polyoxyethylene polyoxypropylene alkyl ether such as olefin glycol ether, polyoxyethylene polyoxypropylene propyl ether, and polyoxyethylene polyoxypropylene cetyl ether; polyoxyethylene alkyl such as polyoxyethylene nonylphenyl ether Polyoxyethylene hardened castor oil such as polyphenylene ether castor oil, polyoxyethylene castor oil, polyoxyethylene hardened castor oil (polyoxyethylene hydrogenated castor oil); polyoxyethylene beeswax derivative such as polyoxyethylene sorbitol beeswax; A polyoxyethylene lanolin derivative such as polyoxyethylene lanolin; a polyoxyethylene fatty guanamine, such as a surfactant of 6 to 18 Hydrophilic-Lipophilic Balance (HLB) such as polyoxyethylene octadecylamine.

非離子性界面活性劑可適當地使用1種或2種以上。The nonionic surfactant can be used singly or in combination of two or more kinds.

一實施方式可為聚氧乙烯去水山梨醇脂肪酸酯或聚氧乙烯聚氧丙烯烷基醚，一實施方式為聚山梨醇酯20、21、40、60、80、81或85，及普蘭尼克(pluronic)型界面活性劑，一實施方式為聚山梨醇酯20及80，或波洛莎姆(波洛莎姆188)。One embodiment may be a polyoxyethylene sorbitan fatty acid ester or a polyoxyethylene polyoxypropylene alkyl ether, and an embodiment is a polysorbate 20, 21, 40, 60, 80, 81 or 85, and Planan A pluronic type surfactant, one embodiment is polysorbate 20 and 80, or a Poloxamer (Polosham 188).

且，非離子性界面活性劑具有抑制不溶性異物或不溶性微粒子等形成之作用，於穩定本發明可用之抗人類α9整合素抗體之範圍內，及/或不對穩定化造成影響之範圍內，可含於醫藥組成物中。
非離子性界面活性劑的量，若為具有穩定化本發明可用之抗人類α9整合素抗體之作用之量，則無特別限制。
利用溶媒(例如注射用水)溶解的溶液狀態(液體製劑)時，可為0.001％(w/v)～1％(w/v)，一實施方式為0.01％(w/v)～0.5％(w/v)，一實施方式為0.01％(w/v)～0.1％(w/v)，一實施方式為0.01％(w/v)～0.03％(w/v)。且，前述的上限與下限可依據期望任意地加以組合。Further, the nonionic surfactant has an action of inhibiting the formation of insoluble foreign matter or insoluble fine particles, and may be included in the range of stabilizing the anti-human α9 integrin antibody usable in the present invention, and/or not affecting the stabilization. In pharmaceutical compositions.
The amount of the nonionic surfactant is not particularly limited as long as it has an effect of stabilizing the anti-human α9 integrin antibody usable in the present invention.
When the state of the solution (liquid preparation) dissolved in a solvent (for example, water for injection) is 0.001% (w/v) to 1% (w/v), and in one embodiment, 0.01% (w/v) to 0.5% ( w/v), an embodiment is 0.01% (w/v) to 0.1% (w/v), and an embodiment is 0.01% (w/v) to 0.03% (w/v). Moreover, the aforementioned upper and lower limits may be arbitrarily combined as desired.

本發明之醫藥組成物可提供為將溶液藉由填充於容器後，製作為液體製劑，或，利用冷凍乾燥法及噴霧乾燥法，製成冷凍乾燥製劑或噴霧乾燥製劑等非經口醫藥組成物。較佳係液體製劑。The pharmaceutical composition of the present invention can be prepared by filling a solution into a container to prepare a liquid preparation, or by using a freeze-drying method and a spray drying method to prepare a non-oral pharmaceutical composition such as a freeze-dried preparation or a spray-dried preparation. . It is preferably a liquid preparation.

本發明之醫藥組成物中可依據期望適當地添加懸濁劑、溶解輔助劑、保存劑、防止黏著劑、含硫還原劑、抗氧化劑等醫藥品添加物。In the pharmaceutical composition of the present invention, a pharmaceutical additive such as a suspending agent, a dissolution aid, a preservative, an anti-adhesion agent, a sulfur-containing reducing agent, or an antioxidant can be appropriately added as desired.

懸濁劑可舉出例如甲基纖維素、羥乙基纖維素、***膠、黃耆樹膠粉末、羧甲基纖維素鈉、聚氧乙烯去水山梨醇單月桂酸酯等。Examples of the suspending agent include methyl cellulose, hydroxyethyl cellulose, gum arabic, gum tragacanth powder, sodium carboxymethyl cellulose, polyoxyethylene sorbitan monolaurate, and the like.

溶解輔助劑可舉出例如聚氧乙烯氫化蓖麻油、尼古丁酸醯胺、聚氧乙烯去水山梨醇單月桂酸酯、聚乙二醇、蓖麻油脂酸乙基乙酯等。The dissolution aid may, for example, be polyoxyethylene hydrogenated castor oil, nicotine acid decylamine, polyoxyethylene sorbitan monolaurate, polyethylene glycol, ricinoleic acid ethyl ethyl ester or the like.

保存劑可舉出例如對羥基苯甲酸甲酯、對羥基苯甲酸乙酯、山梨酸、苯酚、甲酚、氯甲酚等。Examples of the preservative include methyl p-hydroxybenzoate, ethyl p-hydroxybenzoate, sorbic acid, phenol, cresol, chlorocresol and the like.

防止黏著劑可舉出例如人類血清白蛋白、卵磷脂、糊精、氧化乙烯氧化丙烯共聚合物、羥丙基纖維素、甲基纖維素、聚氧乙烯氫化蓖麻油、聚乙二醇等。Examples of the anti-adhesive agent include human serum albumin, lecithin, dextrin, ethylene oxide propylene oxide copolymer, hydroxypropyl cellulose, methyl cellulose, polyoxyethylene hydrogenated castor oil, polyethylene glycol, and the like.

含硫還原劑可舉出例如N-乙醯半胱胺酸、N-乙醯高半胱胺酸、硫辛酸、硫代羥乙酸、硫代乙醇胺、硫代甘油、硫代山梨醇、硫代甘油酸及其鹽、硫代硫酸鈉、麩胺基硫、碳數1～7的硫烷酸等具有硫醇基者等。The sulfur-containing reducing agent may, for example, be N-acetyl cysteine, N-acetyl cystein, lipoic acid, thioglycolic acid, thioethanolamine, thioglycerol, thiosorbitol, thio A thiol group such as glyceric acid or a salt thereof, sodium thiosulfate, glutamine-based sulfur, or a sulfuric acid having 1 to 7 carbon atoms.

抗氧化劑可舉出例如甲硫胺酸、赤藻糖酸、二丙基羥基甲苯、丁基羥基茴香醚、α-生育醇、乙酸生育醇、L-抗壞血酸及其鹽、L-抗壞血酸棕櫚酸酯、L-抗壞血酸硬酯酸酯、亞硫酸氫鈉、沒食子酸三胺、沒食子酸丙酯或乙二胺四乙酸(EDTA)、焦磷酸鈉、偏磷酸鈉等螯合劑等。The antioxidant may, for example, be methionine, erythroic acid, dipropylhydroxytoluene, butylhydroxyanisole, α-tocopherol, tocopheryl acetate, L-ascorbic acid and a salt thereof, and L-ascorbyl palmitate. And a chelating agent such as L-ascorbyl stearate, sodium hydrogen sulfite, gallic acid triamine, propyl gallate or ethylenediaminetetraacetic acid (EDTA), sodium pyrophosphate or sodium metaphosphate.

各種醫藥品添加物於可達成本發明期望效果之量之範圍內，適宜適量地使用。例如，液體製劑的滲透壓，可以調節成為與人類血液本質上相同之250～350mOsm為目的而適宜適量的使用。另外，例如醫藥組成物為溶液狀態(液體製劑)時，將製成可投予用黏度為目的而適宜適量的使用。具體而言，如以後述實施例10記載之測定方法為基礎測定黏度時，可以調製為100cps以下的黏度為目的而適宜適量的使用。Various pharmaceutical additives are suitably used in an amount suitable for the amount of the desired effect of the invention. For example, the osmotic pressure of the liquid preparation can be adjusted to a suitable amount for the purpose of 250 to 350 mOsm which is substantially the same as human blood. Further, for example, when the pharmaceutical composition is in a solution state (liquid preparation), it is suitably used in an amount suitable for the purpose of imparting viscosity. Specifically, when the viscosity is measured based on the measurement method described in the above-described Example 10, it can be suitably used in an appropriate amount for the purpose of preparing a viscosity of 100 cps or less.

本發明醫藥組成物之製造方法，可藉由周知的製造方法，製造含有本發明可用之抗人類α9整合素抗體、製藥學許可的緩衝劑，較佳係進而含有胺基酸類、鹽類、糖或糖醇類，及/或非離子性界面活性劑，包含調整pH為5.0～7.0之方法。
另外，本發明醫藥組成物之製造方法，可藉由周知的製造方法，製造含有本發明可用之抗人類α9整合素抗體、組胺酸或其製藥學許可的鹽，包含調整pH為5.0～7.0之方法。In the method for producing a pharmaceutical composition of the present invention, a buffer containing the anti-human α9 integrin antibody useful in the present invention and a pharmaceutically acceptable drug can be produced by a known production method, and preferably contains an amino acid, a salt, and a sugar. Or a sugar alcohol, and/or a nonionic surfactant, comprising a method of adjusting the pH to 5.0 to 7.0.
Further, in the method for producing a pharmaceutical composition of the present invention, a salt containing an anti-human α9 integrin antibody, histidine or a pharmaceutically acceptable salt thereof which can be used in the present invention can be produced by a known production method, and the pH is adjusted to 5.0 to 7.0. The method.

本發明係包含為了製造含有本發明可用之抗人類α9整合素抗體之穩定的醫藥組成物，製藥學上許可的緩衝劑、胺基酸類、鹽類、糖或糖醇類之作為光穩定化劑之使用。
另外，本發明係包含為了製造含有本發明可用之抗人類α9整合素抗體之穩定的醫藥組成物，將乙酸、檸檬酸、磷酸、組胺酸，以及精胺酸、甲硫胺酸、蔗糖、氯化鈉之作為光穩定劑之使用。
針對本發明之於作為光穩定劑使用時，「穩定」、「穩定的醫藥組成物」、「製藥學上許可的緩衝劑」、「胺基酸類、鹽類、糖或糖醇類」等意義，在本發明之醫藥組成物中可直接適用該當說明。
針對本發明之於作為光穩定劑使用時各成分的摻混量、摻混方法等，在本發明之醫藥組成物中可直接適用該當說明。
於本發明之將乙酸、檸檬酸、磷酸、組胺酸以及該等之製藥學上許可的鹽、精胺酸、甲硫胺酸、蔗糖、氯化鈉之作為光穩定劑之使用，於提供含有本發明可用之抗人類α9整合素抗體之醫藥組成物的同時，可抑制因曝光所導致的該抗人類α9整合素抗體之分解物，或者聚合物的生成。The present invention comprises a pharmaceutically acceptable buffer, an amino acid, a salt, a sugar or a sugar alcohol as a photostabilizer for the manufacture of a stable pharmaceutical composition containing an anti-human α9 integrin antibody useful in the present invention. Use.
Further, the present invention encompasses acetic acid, citric acid, phosphoric acid, histidine, and arginine, methionine, sucrose, in order to produce a stable pharmaceutical composition containing the anti-human α9 integrin antibody useful in the present invention. The use of sodium chloride as a light stabilizer.
For the purpose of using the present invention as a light stabilizer, "stable", "stable pharmaceutical composition", "pharmaceutically acceptable buffer", "amino acid, salt, sugar or sugar alcohol" The description can be directly applied to the pharmaceutical composition of the present invention.
The above description can be directly applied to the pharmaceutical composition of the present invention in the case where the blending amount of each component, the blending method, and the like are used in the present invention as a light stabilizer.
In the present invention, acetic acid, citric acid, phosphoric acid, histidine, and the pharmaceutically acceptable salts, arginine, methionine, sucrose, and sodium chloride are used as light stabilizers. The pharmaceutical composition containing the anti-human α9 integrin antibody usable in the present invention can suppress the decomposition of the anti-human α9 integrin antibody or the formation of a polymer due to exposure.

本發明係包含含有本發明可用之抗人類α9整合素抗體之醫藥組成物中，利用製藥學上許可的緩衝劑、胺基酸類、鹽類、糖或糖醇類，抑制因曝光導致抗人類α9整合素抗體之分解物或聚合物生成之方法。
另外，本發明係包含含有本發明可用之抗人類α9整合素抗體之醫藥組成物中，藉由乙酸、檸檬酸、磷酸、組胺酸以及該等之製藥學上許可的鹽、精胺酸、甲硫胺酸、蔗糖、氯化鈉，抑制因曝光所導致的該抗人類α9整合素抗體之分解物或聚合物的生成之方法。
抑制因曝光所導致的本發明可用之抗人類α9整合素抗體之分解物或聚合物的生成之本發明方法中，使用的「製藥學上許可的緩衝劑」、「胺基酸類、鹽類、糖或糖醇類」、「分解物」、「聚合物」等意義，在本發明之醫藥組成物中可直接適用該當說明。
針對抑制因曝光所導致的本發明可用之抗人類α9整合素抗體之分解物或聚合物的生成之本發明方法中，各成分的摻混量、摻混方法等，在本發明之醫藥組成物中可直接適用該當說明。The present invention comprises a pharmaceutical composition comprising an anti-human α9 integrin antibody useful in the present invention, which inhibits exposure to human α9 by exposure using a pharmaceutically acceptable buffer, an amino acid, a salt, a sugar or a sugar alcohol. A method of decomposing an integrin antibody or a polymer.
Further, the present invention encompasses a pharmaceutical composition comprising an anti-human α9 integrin antibody useful in the present invention, which comprises acetic acid, citric acid, phosphoric acid, histidine, and pharmaceutically acceptable salts thereof, arginine, Methionine, sucrose, and sodium chloride inhibit a method of producing a decomposition product or a polymer of the anti-human α9 integrin antibody by exposure.
The pharmaceutically acceptable buffer, "amino acid, salt," used in the method of the present invention for inhibiting the formation of a decomposition product or a polymer of the anti-human α9 integrin antibody which can be used in the present invention by exposure. The meanings of the sugars, sugar alcohols, "decomposition products", "polymers" and the like can be directly applied to the pharmaceutical compositions of the present invention.
In the method of the present invention for suppressing the formation of a decomposition product or a polymer of an anti-human α9 integrin antibody which can be used in the present invention due to exposure, a blending amount of each component, a blending method, and the like, the pharmaceutical composition of the present invention The description can be directly applied.

填充本發明醫藥組成物的容器可因應使用目的加以選擇。例如包含藥瓶、安瓶、如注射器般一定容量形狀者、如瓶般大容量形狀者。一實施方式係包含注射器(包含拋棄式注射器)。藉由預先將溶液填充於該注射器，提供為填充含藥注射器液體製劑，使臨床醫療無需溶解操作等操作，可迅速對應需求。The container filling the pharmaceutical composition of the present invention can be selected depending on the purpose of use. For example, it includes a medicine bottle, an ampoule, a shape of a certain shape like a syringe, and a large-capacity shape such as a bottle. One embodiment includes a syringe (including a disposable syringe). By filling the solution in advance with the syringe, it is provided to fill the drug-containing syringe liquid preparation, so that the clinical medical operation does not require a dissolution operation or the like, and the demand can be quickly met.

針對容器的材質可舉出玻璃、塑膠等。另外，容器內的表面處理可施以矽被覆處理、二氧化矽被覆處理、硫磺處理、各種低鹼處理等。

[實施例]The material of the container may be glass or plastic. Further, the surface treatment in the container may be subjected to a ruthenium coating treatment, a cerium oxide coating treatment, a sulfur treatment, various low alkali treatments, and the like.

[Examples]

以下，利用參考例、實施例進而詳細說明本發明，但本發明未因該等例示而被限定解釋。Hereinafter, the present invention will be described in detail by way of Reference Examples and Examples. However, the invention is not limited by the examples.

＜＜參考例：抗人類α9整合素抗體之製作＞＞
分別將編碼本實施例中使用之抗人類α9整合素抗體重鏈之聚核苷酸的鹼基序列示於序列編號2，將利用該鹼基序列編碼之胺基酸序列示於序列編號1，編碼抗人類α9整合素抗體輕鏈之聚核苷酸的鹼基序列示於序列編號4，將利用該鹼基序列編碼之胺基酸序列示於序列編號3。<<Reference example: Production of anti-human α9 integrin antibody>>
The nucleotide sequence of the polynucleotide encoding the heavy chain of the anti-human α9 integrin antibody used in the present Example is shown in SEQ ID NO: 2, and the amino acid sequence encoded by the nucleotide sequence is shown in SEQ ID NO: 1, The nucleotide sequence of the polynucleotide encoding the light chain of the anti-human α9 integrin antibody is shown in SEQ ID NO: 4, and the amino acid sequence encoded by the base sequence is shown in SEQ ID NO: 3.

遵循國際公開第2009/088064號公報，構築***有該抗人類α9整合素抗體重鏈之AG-γ1 (WO94/20632)，以及***有該抗人類α9整合素抗體輕鏈之AG-κ1(WO94/20632)，並製作連結該等之表現質體。使用FreeStyle 293 Expression System使該表現質體表現，再使用蛋白質A管柱自培養上清液取得純化抗體。以及，分析經純化之抗人類α9整合素抗體之胺基酸修飾後，發現其大部分的重鏈N端產生焦麩胺醯化，以及產生重鏈C端側的離胺酸的缺失。AG-γ1 (WO94/20632) inserted with the anti-human α9 integrin antibody heavy chain, and AG-κ1 (WO94) inserted with the anti-human α9 integrin antibody light chain were constructed in accordance with International Publication No. 2009/088064. /20632), and create a representation of the quality of the body. The expression of the plastid was expressed using a FreeStyle 293 Expression System, and the purified antibody was obtained from the culture supernatant using a Protein A column. Further, after analysis of the amino acid modified by the purified anti-human α9 integrin antibody, it was found that most of the heavy chain N-terminus produced pyroguanamine deuteration, and the amino acid-depleted C-terminal side was deficient in the amino acid.

＜＜實施例1：藉由選定最適pH之穩定化效果＞＞
將利用依據表1所示緩衝液稀釋，使依參考例製造之抗人類α9整合素抗體濃度成為10mg/mL所調製之處方液(試料No. A-1～A-7)，以0.22μm過濾器進行滅菌過濾後，以各1.3mL填充至玻璃藥瓶(3mL容量)，再以橡膠拴進行打拴，以正置狀態於40℃保存1個月，以及，對利用表1所示緩衝液所稀釋調製之處方液中之試料No. A-1、2、3、6、7，以D65燈實施1000lux，48小時的曝光。且將以D65燈曝光的檢體作為對照檢體，以鋁箔紙包覆方法製作為遮光檢體亦進行保存，並評價其穩定性。
另外，為使其成為預期的pH，可因應需要將作為pH調節劑之鹽酸及/或氫氧化鈉，於調製緩衝液時進行添加。<<Example 1: Stabilization effect by selecting optimum pH>>
The concentration of the anti-human α9 integrin antibody produced according to the reference example was adjusted to 10 mg/mL (sample No. A-1 to A-7), and the mixture was filtered at 0.22 μm by dilution with the buffer shown in Table 1. After sterilization and filtration, the cells were filled into a glass vial (3 mL capacity) with 1.3 mL each, and then snored with a rubber crucible, stored at 40 ° C for 1 month in an upright state, and the buffer shown in Table 1 was used. Sample No. A-1, 2, 3, 6, and 7 in the diluted solution were subjected to 1000 lux for 48 hours with a D65 lamp. Further, the sample exposed by the D65 lamp was used as a control sample, and a light-shielding sample was prepared by an aluminum foil coating method, and the stability was evaluated.
Further, in order to obtain a desired pH, hydrochloric acid and/or sodium hydroxide as a pH adjuster may be added as needed in the preparation of a buffer.

SE-HPLC測定係將HPLC系統與TSKgel G3000SWXL管柱(5μm；7.8mm×300mm，TOSOH製)連結，並以50 mmol/L磷酸/300mmol/L氯化鈉(pH6.8)組成之移動相，以0.5mL/分鐘流速沖流。將測定檢體的濃度定為10mg/mL，以注入量400μg條件進行分析。將管柱溫度設定於25℃，UV測定的波長使用280nm。
測定以SE-HPLC測定檢出之波峰面積，藉由除以包含主要波峰之全波峰面積的總和，計算為百分率(％)。且，主要波峰係意指活性本體波峰。
合併較SE-HPLC主要波峰的維持時間更短的波峰作為聚合物量，另合併維持時間較長的波峰作為分解物量。另外，將聚合物及分解物的總稱規定為不純物時，以聚合物量及分解物量的總和作為不純物量。結果示於圖1～圖4。
藉由將本抗人類α9整合素抗體於pH5.0～7.0附近進行製劑化，顯示可以抑制聚合物、分解物之生成。The SE-HPLC assay was performed by linking an HPLC system to a TSKgel G3000SWXL column (5 μm; 7.8 mm × 300 mm, manufactured by TOSOH) and a mobile phase consisting of 50 mmol/L phosphoric acid/300 mmol/L sodium chloride (pH 6.8). The flow was flowed at a flow rate of 0.5 mL/min. The concentration of the test sample was set to 10 mg/mL, and the analysis was carried out under the conditions of an injection amount of 400 μg. The column temperature was set to 25 ° C, and the wavelength of the UV measurement was 280 nm.
The peak area detected by SE-HPLC measurement was measured and divided into percentage (%) by dividing by the sum of the full peak areas including the main peaks. Moreover, the main peak system means the active body peak.
A peak having a shorter retention time than the main peak of the SE-HPLC was combined as the amount of the polymer, and a peak having a longer maintenance time was combined as the amount of the decomposition product. Further, when the general term of the polymer and the decomposed product is defined as an impurity, the sum of the amount of the polymer and the amount of the decomposed product is regarded as the amount of the impurity. The results are shown in Figs. 1 to 4 .
Formulation of the anti-human α9 integrin antibody at a pH of 5.0 to 7.0 shows that formation of a polymer or a decomposed product can be suppressed.

＜＜實施例2：藉由選定緩衝劑成分之穩定化效果＞＞
表2係揭示含有參考例所得10mg/mL抗人類α9整合素抗體、緩衝液(20mmol/L檸檬酸(pH6.0)、20mmol/L磷酸(pH6.0)、或20mmol/L組胺酸(pH6.0))之處方(試料No. B-1～B-3)。且，為使其成為預期的pH可因應需要將作為pH調節劑之鹽酸及/或氫氧化鈉，於調製緩衝液時進行添加。將依該緩衝液稀釋調製之處方液分別以0.22μm過濾器進行滅菌過濾後，以1.3mL填充至各玻璃藥瓶(3mL容量)，再以橡膠拴進行打拴，以正置狀態於40℃保存1個月，及實施以D65燈實施1000lux，48小時的曝光。且將作為D65燈曝光檢體的對照檢體，以鋁箔紙包覆方法製作為遮光檢體亦進行保存，並評價其穩定性。<<Example 2: Stabilization effect by selecting a buffer component>>
Table 2 discloses the 10 mg/mL anti-human α9 integrin antibody obtained in the reference example, a buffer solution (20 mmol/L citric acid (pH 6.0), 20 mmol/L phosphoric acid (pH 6.0), or 20 mmol/L histidine ( pH 6.0)) (sample No. B-1 to B-3). Further, in order to make it a desired pH, it is necessary to add hydrochloric acid and/or sodium hydroxide as a pH adjuster in the preparation of a buffer. The solution prepared by diluting the buffer solution was sterilized and filtered with a 0.22 μm filter, and then filled into each glass vial (3 mL capacity) with 1.3 mL, and then snored with a rubber crucible, and placed at 40 ° C in an upright state. Store for 1 month, and perform 1000 lux with 48 hours exposure with D65 lamp. Further, a control sample which was a D65 lamp exposure sample was prepared as a light-shielding specimen by an aluminum foil coating method, and its stability was evaluated.

評價抗人類α9整合素抗體的穩定性(聚合物量、分解物量)。結果示於圖5～圖7。
聚合物量、分解物量係依據SE-HPLC法，與實施例1進行相同的分析。
於40℃保存1個月及以D65燈照射下保存後的檢體中，組胺酸最可抑制聚合物的生成。且，含有檸檬酸及磷酸的檢體任一個均屬穩定。The stability (amount of polymer, amount of decomposition product) of the anti-human α9 integrin antibody was evaluated. The results are shown in Figs. 5 to 7 .
The amount of the polymer and the amount of the decomposition product were the same as those in Example 1 according to the SE-HPLC method.
The histidine inhibited the formation of the polymer most in the sample stored at 40 ° C for one month and stored under a D65 lamp. Moreover, any of the samples containing citric acid and phosphoric acid is stable.

＜＜實施例3：藉由選定緩衝劑成分之長期曝光後穩定化效果＞＞
表3係揭示含有參考例所得10mg/mL抗人類α9整合素抗體、緩衝液(20mmol/L組胺酸(pH6.0)、100mmol/L組胺酸(pH6.0)、20mmol/L檸檬酸(pH6.0)、20mmol/L磷酸(pH6.0)、20mmol/L琥珀酸(pH6.0)、20mmol/L2-嗎林代乙烷磺酸(以下稱作MES) (pH6.0)、20mmol/L酒石酸(pH6.0))之處方(試料No. B-4～B-10)。且，為使其成為預期的pH可因應需要將作為pH調節劑之鹽酸及/或氫氧化鈉，於調製緩衝液時進行添加。將依該緩衝液稀釋調製之處方液分別以0.22μm過濾器進行滅菌過濾後，以0.5mL填充至各玻璃藥瓶(3mL容量)，再以橡膠拴進行打拴，實施以D65燈實施1000lux，168小時的曝光。且將對照檢體，以鋁箔紙包覆方法製作為遮光檢體亦進行保存，並評價其穩定性。<<Example 3: Stabilization effect after long-term exposure by selecting a buffer component>>
Table 3 discloses the 10 mg/mL anti-human α9 integrin antibody obtained in the reference example, buffer (20 mmol/L histidine (pH 6.0), 100 mmol/L histidine (pH 6.0), 20 mmol/L citric acid). (pH 6.0), 20 mmol/L phosphoric acid (pH 6.0), 20 mmol/L succinic acid (pH 6.0), 20 mmol/L 2-morphinated ethanesulfonic acid (hereinafter referred to as MES) (pH 6.0), 20 mmol/L tartaric acid (pH 6.0)) (sample No. B-4 to B-10). Further, in order to make it a desired pH, it is necessary to add hydrochloric acid and/or sodium hydroxide as a pH adjuster in the preparation of a buffer. The solution prepared by diluting the buffer solution was sterilized and filtered with a 0.22 μm filter, and then filled into each glass vial (3 mL capacity) with 0.5 mL, and then rubbed with a rubber crucible to carry out 1000 lux with a D65 lamp. 168 hours of exposure. The control sample was also prepared as a light-shielding specimen by an aluminum foil coating method, and its stability was evaluated.

評價參考例獲得之抗人類α9整合素抗體的穩定性(聚合物量)。結果示於圖8。聚合物量係依據SE-HPLC法，與實施例1進行相同的分析。
檢證緩衝劑種類中，組胺酸最可抑制聚合物的生成。且，組胺酸濃度愈高，愈可抑制聚合物的生成。The stability (polymer amount) of the anti-human α9 integrin antibody obtained in the reference example was evaluated. The results are shown in Figure 8. The amount of the polymer was analyzed in the same manner as in Example 1 according to the SE-HPLC method.
Among the types of test buffers, histidine most inhibits the formation of polymers. Moreover, the higher the concentration of histidine, the more the formation of the polymer can be inhibited.

＜＜實施例4：藉由選定胺基酸類、鹽類、糖或糖醇類之穩定化效果＞＞
於含有參考例所得10mg/mL抗人類α9整合素抗體、20mmol/L檸檬酸(pH6.0)之處方，不添加添加劑，或是添加精胺酸(20mmol/L、100mmol/L或200mmol/L)、氯化鈉(NaCl，20mmol/L、100mmol/L或200mmol/L)、甲硫胺酸(10mmol/L、20mmol/L或100mmol/L)、山梨糖醇(200mmol/L)，或是蔗糖(200mmol/L)。且，為使其成為預期的pH可因應需要將作為pH調節劑之鹽酸及/或氫氧化鈉，於調製緩衝液時進行添加。處方液分別以0.22μm過濾器進行滅菌過濾後，以1.3mL填充至各玻璃藥瓶(3mL容量)，再以橡膠拴進行打拴，以正置狀態於40℃保存1個月。
另外，針對含有精胺酸、甲硫胺酸、氯化鈉、山梨糖醇或蔗糖的檢體，以D65燈實施1000lux、48小時的曝光。且將與D65燈保存之檢體作為對照檢體，以鋁箔紙包覆方法製作為遮光檢體亦進行保存，並評價其穩定性。<<Example 4: Stabilization effect by selecting an amino acid, a salt, a sugar or a sugar alcohol>>
Including the 10 mg/mL anti-human α9 integrin antibody obtained in the Reference Example and 20 mmol/L citric acid (pH 6.0), no additive or arginine (20 mmol/L, 100 mmol/L or 200 mmol/L) was added. ), sodium chloride (NaCl, 20mmol/L, 100mmol/L or 200mmol/L), methionine (10mmol/L, 20mmol/L or 100mmol/L), sorbitol (200mmol/L), or Sucrose (200 mmol/L). Further, in order to make it a desired pH, it is necessary to add hydrochloric acid and/or sodium hydroxide as a pH adjuster in the preparation of a buffer. The prescription liquid was sterilized and filtered with a 0.22 μm filter, and then filled into each glass vial (3 mL capacity) with 1.3 mL, and then rubbed with a rubber crucible, and stored at 40 ° C for 1 month in an upright state.
Further, for a sample containing arginine, methionine, sodium chloride, sorbitol or sucrose, 1000 lux and 48 hours of exposure were carried out with a D65 lamp. Further, the sample stored in the D65 lamp was used as a control sample, and a light-shielding sample was also prepared by an aluminum foil coating method, and the stability was evaluated.

聚合物量係依據SE-HPLC法，與實施例1進行相同的分析。
結果示於圖9～圖14。
檢討後發現精胺酸、氯化鈉(NaCl)、甲硫胺酸、山梨糖醇或蔗糖，可使抗人類α9整合素抗體於40℃保存條件下穩定化，或是不會對穩定性造成影響。
針對精胺酸、氯化鈉(NaCl)、甲硫胺酸、山梨糖醇或蔗糖，顯示對光有穩定化效果，或者對光呈現穩定。The amount of the polymer was analyzed in the same manner as in Example 1 according to the SE-HPLC method.
The results are shown in Figs. 9 to 14 .
After review, it was found that arginine, sodium chloride (NaCl), methionine, sorbitol or sucrose can stabilize anti-human α9 integrin antibody at 40 ° C or not. influences.
For arginine, sodium chloride (NaCl), methionine, sorbitol or sucrose, it has a stabilizing effect on light or is stable to light.

＜＜實施例5：藉由添加非離子性界面活性劑之穩定化效果＞＞
於含有參考例所得10mg/mL抗人類α9整合素抗體、20mmol/L組胺酸(pH6.0)、140mmol/L精胺酸之處方，分別以0、0.005、0.01、0.02、0.05或0.1w/v％來添加非離子性界面活性劑之聚山梨醇酯80。且，為使其成為預期的pH可因應需要將作為pH調節劑之鹽酸及/或氫氧化鈉，於調製緩衝液時進行添加。處方液分別以0.22μm過濾器進行滅菌過濾後，以4.4mL填充至各玻璃藥瓶(10mL容量)，再以橡膠拴進行打拴，評價對於振盪、冷凍融解壓力的物理性的穩定性。
振盪試驗係將水平放置的玻璃藥瓶以150rpm震盪24小時。
冷凍融解壓力試驗係將檢體以正置狀態保存於-80℃之冷凍庫內使其冷凍後，再將檢體自冷凍庫中取出，於5℃的冷藏庫以正置狀態保存並使其融解。重複3次進行該冷凍融解循環。<<Example 5: Stabilization effect by adding a nonionic surfactant>>
10mg/mL anti-human α9 integrin antibody, 20mmol/L histidine (pH6.0), 140mmol/L arginine obtained in the reference example, respectively, 0, 0.005, 0.01, 0.02, 0.05 or 0.1w /v% is added to the non-ionic surfactant polysorbate 80. Further, in order to make it a desired pH, it is necessary to add hydrochloric acid and/or sodium hydroxide as a pH adjuster in the preparation of a buffer. The prescription liquid was sterilized and filtered by a 0.22 μm filter, and then filled into each glass vial (10 mL capacity) with 4.4 mL, and then rubbed with a rubber crucible to evaluate the physical stability against the oscillation and freezing melt pressure.
The shaking test was to oscillate the horizontally placed glass vial at 150 rpm for 24 hours.
In the freeze-thaw pressure test, the sample was stored in a freezer at -80 ° C in an upright state, and then frozen, and then the sample was taken out from the freezer, and stored in a refrigerator at 5 ° C in an upright state and allowed to melt. This freeze-thaw cycle was repeated 3 times.

利用光遮蔽粒子計數法的評價結果示於圖15及圖16。
光遮蔽粒子計數法係使用液中粒子計數器(HIAC Lab型液中粒子計數器)進行測定。將測定檢體的濃度定為10mg/mL，75Torr，25℃，2小時靜置條件脫氣後，以注入量0.2mL條件進行分析。
利用光遮蔽粒子計數法，測定檢出粒子徑1.2μm以上的不溶性微粒子數，以1mL作為換算值進行計算。
結果顯示，抗人類α9整合素抗體於含有非離子性界面活性劑聚山梨醇酯80的製劑中，可抑制因冷凍融解、震盪而生成的不溶性微粒子。The evaluation results by the light shielding particle counting method are shown in Fig. 15 and Fig. 16 .
The light-shielding particle counting method was measured using a liquid particle counter (HIAC Lab type liquid particle counter). The concentration of the test sample was set to 10 mg/mL, 75 Torr, and 25 ° C, and after degassing for 2 hours, the analysis was carried out under the conditions of an injection amount of 0.2 mL.
The number of insoluble fine particles having a particle diameter of 1.2 μm or more was measured by a light-shielding particle counting method, and was calculated by using 1 mL as a converted value.
As a result, it was revealed that the anti-human α9 integrin antibody can inhibit insoluble fine particles generated by freezing and oscillating in a preparation containing the nonionic surfactant polysorbate 80.

＜＜實施例6：檢討含組胺酸、精胺酸、聚山梨醇酯80處方之穩定性評價＞＞
於含有參考例所得10mg/mL抗人類α9整合素抗體、20mmol/L組胺酸、140mmol/L精胺酸、0.02 w/v％聚山梨醇酯80之處方液，以0.22μm過濾器進行滅菌過濾後，分別以4.4mL填充至各玻璃藥瓶(10mL容量)，再以橡膠拴進行打拴，對於振盪、冷凍融解、光、保存於-20℃、5℃及25℃時的穩定性，利用SE-HPLC法、陽離子交換管柱層析法(CE-HPLC法)、光遮蔽粒子計數法進行評價。
振盪、冷凍融解壓力試驗係以與實施例5相同方法進行。
光穩定性試驗係以與實施例2相同方法進行。
保存於-20℃、5℃及25℃時的穩定性試驗係以與實施例1相同方法進行。
且，為使其成為預期的pH(pH6.0)，可因應需要於調製緩衝液時添加作為pH調節劑之鹽酸及/或氫氧化鈉。<<Example 6: Review of stability evaluation of prescription containing histidine, arginine and polysorbate 80>>
The solution containing 10 mg/mL anti-human α9 integrin antibody, 20 mmol/L histidine, 140 mmol/L arginine, 0.02 w/v% polysorbate 80 obtained in the reference example was sterilized by a 0.22 μm filter. After filtration, each of the glass vials (10 mL capacity) was filled with 4.4 mL, and then rubbed with a rubber crucible for stability of shaking, freezing, melting, light, and storage at -20 ° C, 5 ° C, and 25 ° C. The evaluation was carried out by SE-HPLC method, cation exchange column chromatography (CE-HPLC method), and light-shielding particle counting method.
The shaking and freezing melt pressure test was carried out in the same manner as in Example 5.
The light stability test was carried out in the same manner as in Example 2.
The stability test at -20 ° C, 5 ° C and 25 ° C was carried out in the same manner as in Example 1.
Further, in order to make it a desired pH (pH 6.0), hydrochloric acid and/or sodium hydroxide as a pH adjuster may be added as needed in the preparation of the buffer.

SE-HPLC法係以與實施例1相同方法進行，光遮蔽粒子計數法係以與實施例5相同方法進行測定。
CE-HPLC法測定係HPLC系統與ProPac WCX-10管柱(10μm；4mm×250mm，ThermoScientific製)連結，移動相A：20mmol/L的MES(2-(N-morpholino)ethanesulfonic acid) pH6.0，移動相B：20mmol/L的MES/500 mmol/L氯化鈉pH6.0之組成的移動相，並以0.5mL/分鐘流速及以表4所示梯度條件沖流。將測定檢體的濃度定為1mg/mL，以注入量10μg條件進行分析。將管柱溫度設定於40℃，UV測定的波長使用280nm。The SE-HPLC method was carried out in the same manner as in Example 1, and the light-shielding particle counting method was carried out in the same manner as in Example 5.
CE-HPLC method HPLC system was coupled with ProPac WCX-10 column (10 μm; 4 mm × 250 mm, manufactured by Thermo Scientific), mobile phase A: 20 mmol/L MES (2-(N-morpholino) ethanesulfonic acid) pH 6.0 , mobile phase B: mobile phase consisting of 20 mmol/L MES/500 mmol/L sodium chloride pH 6.0, and flowed at a flow rate of 0.5 mL/min and under the gradient conditions shown in Table 4. The concentration of the test sample was set to 1 mg/mL, and the analysis was carried out under the conditions of an injection amount of 10 μg. The column temperature was set to 40 ° C, and the wavelength of the UV measurement was 280 nm.

測定以CE-HPLC測定檢出之波峰面積，藉由除以包含波峰1(主要波峰)之全波峰面積的總和，計算為百分率(％)。且，主要波峰係意指活性本體顯示之最大面積的波峰。
以CE-HPLC測定檢出之波峰內，將保存中變動較大的波峰定為特定波峰2、波峰3。
結果示於表5及表6。
該處方係穩定的狀況。The peak area detected by CE-HPLC measurement was measured and divided into percentage (%) by dividing by the sum of the full peak areas including peak 1 (main peak). Moreover, the main peak system means the peak of the largest area of the active body display.
In the peak detected by the CE-HPLC measurement, the peak with a large fluctuation in storage was designated as a specific peak 2 and a peak 3.
The results are shown in Tables 5 and 6.
This prescription is a stable condition.

＜＜實施例7：檢討含組胺酸、精胺酸、聚山梨醇酯80之高濃度液體製劑之穩定性評價＞＞
於含有參考例所得100mg/mL抗人類α9整合素抗體、20mg/L組胺酸(pH6.0)、140mg/L精胺酸、0.02 w/v％聚山梨醇酯80之處方，調節pH為5.0、5.5、6.0等3個程度。將處方液分別以0.22μm過濾器進行滅菌過濾後，再分別以0.4mL或0.3mL填充至微管中。
且，為使其成為預期的pH可因應需要將作為pH調節劑之鹽酸及/或氫氧化鈉，於調製緩衝液時進行添加。<<Example 7: Evaluation of stability evaluation of high concentration liquid preparation containing histidine, arginine and polysorbate 80>>
The pH was adjusted to include 100 mg/mL anti-human α9 integrin antibody obtained in Reference Example, 20 mg/L histidine (pH 6.0), 140 mg/L arginine, and 0.02 w/v% polysorbate 80. 5.0, 5.5, 6.0, etc. 3 degrees. The prescription liquids were sterilized and filtered by a 0.22 μm filter, and then filled into microtubes at 0.4 mL or 0.3 mL, respectively.
Further, in order to make it a desired pH, it is necessary to add hydrochloric acid and/or sodium hydroxide as a pH adjuster in the preparation of a buffer.

於25℃保存1個月時的穩定性係利用SE-HPLC法進行評價。
SE-HPLC法係以與實施例1相同方法測定。
結果示於圖17～圖19。
於各pH均係穩定的狀況。The stability at 1 month storage at 25 ° C was evaluated by the SE-HPLC method.
The SE-HPLC method was measured in the same manner as in Example 1.
The results are shown in Figs. 17 to 19 .
It is stable at each pH.

＜＜實施例8：藉由選定胺基酸類、糖或糖醇類之高濃度液體製劑之穩定化效果＞＞
於含有參考例所得100mg/mL抗人類α9整合素抗體、20mg/L組胺酸(pH6.0)之處方，分別將經添加20mg/L甲硫胺酸、280mg/L山梨糖醇、280mg/L蔗糖、或140mg/L精胺酸之檢體，以0.22μm過濾器進行滅菌過濾後，再分別以0.4mL或0.3mL填充至微管中。
且，為使其成為預期的pH可因應需要將作為pH調節劑之鹽酸及/或氫氧化鈉，於調製緩衝液時進行添加。<<Example 8: Stabilization effect by selecting a high concentration liquid preparation of an amino acid, a sugar or a sugar alcohol>>
After adding 100 mg/mL anti-human α9 integrin antibody and 20 mg/L histidine (pH 6.0) obtained in the reference example, 20 mg/L methionine, 280 mg/L sorbitol, 280 mg/, respectively, were added. The sample of L sucrose or 140 mg/L arginine was sterilized and filtered with a 0.22 μm filter, and then filled into microtubes with 0.4 mL or 0.3 mL, respectively.
Further, in order to make it a desired pH, it is necessary to add hydrochloric acid and/or sodium hydroxide as a pH adjuster in the preparation of a buffer.

於25℃保存1個月時的穩定性係利用SE-HPLC法進行評價。
SE-HPLC法係以與實施例1相同方法測定。
結果示於圖20～圖22。
各含有醫藥添加物的製劑均係穩定的狀況。The stability at 1 month storage at 25 ° C was evaluated by the SE-HPLC method.
The SE-HPLC method was measured in the same manner as in Example 1.
The results are shown in Figs. 20 to 22 .
Each of the preparations containing the pharmaceutical additive was in a stable state.

＜＜實施例9：藉由添加非離子性界面活性劑之高濃度液體製劑之穩定化效果＞＞
於含有參考例所得100mg/mL抗人類α9整合素抗體、20mg/L組胺酸(pH6.0)、140mg/L精胺酸之處方，將經添加0.02 w/v％聚山梨醇酯80或波洛莎姆188之處方液，以0.22μm過濾器進行滅菌過濾後，再以各1.2mL填充至3mL的玻璃藥瓶中。且，為使其成為預期的pH可因應需要將作為pH調節劑之鹽酸及/或氫氧化鈉，於調製緩衝液時進行添加。<<Example 9: Stabilization effect of a high concentration liquid preparation by adding a nonionic surfactant>>
Add 0.02 w/v% polysorbate 80 or 80 mg/mL anti-human α9 integrin antibody, 20 mg/L histidine (pH 6.0), 140 mg/L arginine. The solution of Polosham 188 was sterilized and filtered with a 0.22 μm filter, and then filled into a 3 mL glass vial with 1.2 mL each. Further, in order to make it a desired pH, it is necessary to add hydrochloric acid and/or sodium hydroxide as a pH adjuster in the preparation of a buffer.

對振盪、冷凍融解壓力進行物理的穩定性評價。
振盪試驗以及冷凍融解壓力試驗係以與實施例5相同方法進行。
利用光遮蔽粒子計數法的評價結果示於圖23及圖24。
光遮蔽粒子計數法係以與實施例5相同方法進行。
聚山梨醇酯80、波洛莎姆188均抑制不溶性微粒子的生成。Physical stability evaluation of the oscillation and freezing melt pressure was performed.
The oscillation test and the freeze-thaw pressure test were carried out in the same manner as in Example 5.
The evaluation results by the light shielding particle counting method are shown in FIGS. 23 and 24.
The light-shielding particle counting method was carried out in the same manner as in Example 5.
Both polysorbate 80 and Poloxamer 188 inhibit the formation of insoluble microparticles.

＜＜實施例10：檢討含有組胺酸、精胺酸、波洛莎姆188之高濃度液體製劑之穩定性評價＞＞
於含有參考例所得100mg/mL～200mg/mL抗人類α9整合素抗體、90mg/L組胺酸(pH5.8)、240mg/L精胺酸、0.02 w/v％波洛莎姆188之處方，將處方液分別以0.22μm過濾器進行滅菌過濾後，再以各1.3mL填充至3mL的玻璃藥瓶中，並以橡膠拴打拴。
且，為使其成為預期的pH可因應需要將作為pH調節劑之鹽酸及/或氫氧化鈉，於調製緩衝液時進行添加。<<Example 10: Evaluation of stability evaluation of liquid preparation containing high concentration of histidine, arginine and polosa 188>>
Containing 100 mg/mL to 200 mg/mL anti-human α9 integrin antibody, 90 mg/L histidine (pH 5.8), 240 mg/L arginine, 0.02 w/v% Poloxaram 188 obtained in the reference example The prescription liquid was sterilized and filtered by a 0.22 μm filter, and then filled into a 3 mL glass vial with 1.3 mL each, and smashed with a rubber crucible.
Further, in order to make it a desired pH, it is necessary to add hydrochloric acid and/or sodium hydroxide as a pH adjuster in the preparation of a buffer.

於5℃、25℃保存3個月時的穩定性係利用SE-HPLC法進行評價。
SE-HPLC法係以與實施例1相同方法測定。
黏度測定係將落球式黏度計(Automated Micro Viscometer, Anton Paar製)與毛細管(內徑1.2mm, Anton Paar製)連結，使用金屬製球(直徑1.0mm, Anton Paar製)藉由使球落下而測定。設定毛細管溫度為20℃，測定角度為70°，密度為1.00000g/cm³ 並進行密度測定。
結果示於表7。
於各抗體濃度該處方均係穩定。The stability at 3 ° C and 25 ° C for 3 months was evaluated by the SE-HPLC method.
The SE-HPLC method was measured in the same manner as in Example 1.
For the viscosity measurement, a ball-type viscometer (Automated Micro Viscometer, manufactured by Anton Paar) was connected to a capillary tube (inner diameter: 1.2 mm, manufactured by Anton Paar), and a metal ball (1.0 mm in diameter, manufactured by Anton Paar) was used to drop the ball. Determination. The capillary temperature was set to 20 ° C, the measurement angle was 70 °, the density was 1.00000 g/cm ^{3 ,} and the density was measured.
The results are shown in Table 7.
The formulation is stable at each antibody concentration.

＜＜實施例11：檢討含有組胺酸、精胺酸、波洛莎姆188之高濃度液體製劑之穩定性評價＞＞
將含有參考例所得之抗人類α9整合素抗體及以表8所示之處方液，分別以0.22μm過濾器進行滅菌過濾後，再以各1.3mL填充至3mL的玻璃藥瓶中，並以橡膠拴打拴。
且，為使其成為預期的pH可因應需要將作為pH調節劑之鹽酸及/或氫氧化鈉，於調製緩衝液時進行添加。<<Example 11: Evaluation of stability evaluation of high-concentration liquid preparation containing histidine, arginine, and polosa 188>>
The anti-human α9 integrin antibody obtained in the reference example and the solution shown in Table 8 were sterilized and filtered with a 0.22 μm filter, and then filled into a 3 mL glass vial with 1.3 mL each, and rubber was used. Please call.
Further, in order to make it a desired pH, it is necessary to add hydrochloric acid and/or sodium hydroxide as a pH adjuster in the preparation of a buffer.

於25℃保存3個月時的穩定性係利用SE-HPLC法及CE-HPLC法進行評價。
統計解析係使用JMP-11(SAS Institute Inc. 製)進行解析。
SE-HPLC法係以與實施例1相同方法測定，測定聚合物量及分解物量。
CE-HPLC法係以與實施例6相同方法測定，測定主波峰量。
結果示於圖25～圖27。實線係表示依據獲得的數據為基礎的統計解析而預測之值，實線上下的點線係依據統計解析而預測之值的幅度。
該處方在抗體濃度為200mg/mL時，於各組胺酸濃度、精胺酸濃度均係穩定。

[產業上之可利用性]The stability at 3 hours storage at 25 ° C was evaluated by SE-HPLC method and CE-HPLC method.
The statistical analysis system was analyzed using JMP-11 (manufactured by SAS Institute Inc.).
The SE-HPLC method was measured in the same manner as in Example 1, and the amount of the polymer and the amount of the decomposition product were measured.
The CE-HPLC method was measured in the same manner as in Example 6, and the amount of main peaks was measured.
The results are shown in Figs. 25 to 27 . The solid line indicates the value predicted based on the statistical analysis based on the obtained data, and the dotted line on the solid line is the magnitude of the value predicted based on the statistical analysis.
When the antibody concentration was 200 mg/mL, the prescription was stable at each histidine concentration and arginine concentration.

[Industrial availability]

根據本發明可提供含有抗人類α9整合素抗體所構成之穩定的醫藥組成物，詳細係可提供由抑制聚合物、分解物、不溶性異物或不溶性微粒子的生成，含有抗人類α9整合素抗體所構成之穩定的醫藥組成物。
上述內容係依據特定格式說明本發明，但本發明範圍係包含相關業者明確已知之改變作法之方法與改良。

[序列表之非關鍵詞文字]According to the present invention, it is possible to provide a stable pharmaceutical composition comprising an anti-human α9 integrin antibody, which can provide a composition comprising an anti-human α9 integrin antibody by inhibiting the formation of a polymer, a decomposition product, an insoluble foreign matter or insoluble microparticles. Stable pharmaceutical composition.
The above description is based on the specific format of the invention, but the scope of the invention includes methods and improvements that are well known to those skilled in the art.

[Non-keyword text of sequence list]

序列表序列編號1表示之胺基酸序列為抗人類α9整合素抗體之重鏈胺基酸序列。
同序列編號2表示之鹼基酸序列為抗人類α9整合素抗體之重鏈基因的鹼基序列。
同序列編號3表示之胺基酸序列為抗人類α9整合素抗體之輕鏈胺基酸序列。
同序列編號4表示之鹼基酸序列為抗人類α9整合素抗體之輕鏈基因的鹼基序列。The amino acid sequence represented by Sequence Listing SEQ ID NO: 1 is a heavy chain amino acid sequence of an anti-human α9 integrin antibody.
The base acid sequence represented by the same sequence number 2 is the base sequence of the heavy chain gene of the anti-human α9 integrin antibody.
The amino acid sequence represented by the same sequence number 3 is the light chain amino acid sequence of the anti-human α9 integrin antibody.
The base acid sequence represented by the same sequence number 4 is the base sequence of the light chain gene of the anti-human α9 integrin antibody.

[圖1]實施例1進行之將各種pH的液體製劑，於40℃、1個月的保存期間前後之粒徑篩析層析法(SE-HPLC法)的評價結果(聚合物量)之圖。Fig. 1 is a graph showing the results of evaluation (polymer amount) of particle size screening chromatography (SE-HPLC method) of liquid preparations of various pHs carried out in Example 1 before and after storage at 40 ° C for one month. .

[圖2]實施例1進行之將各種pH的液體製劑，於40℃、1個月的保存期間前後之SE-HPLC法的評價結果(分解物量)之圖。 Fig. 2 is a graph showing the results of evaluation by SE-HPLC method (amount of decomposition product) of liquid preparations of various pHs in Example 1 before and after storage at 40 ° C for one month.

[圖3]實施例1進行之將各種pH的液體製劑，於40℃、1個月的保存期間前後之SE-HPLC法的評價結果(不純物(=聚合物+分解物)量)之圖。 Fig. 3 is a graph showing the results of evaluation by SE-HPLC method (amount of impurities (= polymer + decomposed product)) of a liquid preparation of various pHs in Example 1 before and after storage at 40 ° C for one month.

[圖4]實施例1進行之將含有各種緩衝液成分的液體製劑，於48小時曝光前後之SE-HPLC法的評價結果(聚合物量)圖。 Fig. 4 is a graph showing the results of evaluation by SE-HPLC method (polymer amount) of a liquid preparation containing various buffer components in Example 1 before and after exposure for 48 hours.

[圖5]實施例2進行之將含有各種緩衝液成分的液體製劑，於40℃、1個月的保存期間前後之SE-HPLC法的評價結果(聚合物量)圖。 Fig. 5 is a graph showing the results of evaluation by SE-HPLC method (polymer amount) of a liquid preparation containing various buffer components in Example 2 before and after storage at 40 °C for one month.

[圖6]實施例2進行之將含有各種緩衝液成分的液體製劑，於40℃、1個月的保存期間前後之SE-HPLC法的評價結果(分解物量)圖。 Fig. 6 is a graph showing the results of evaluation by SE-HPLC method (amount of decomposition product) of a liquid preparation containing various buffer components in Example 2 before and after storage at 40 °C for one month.

[圖7]實施例2進行之將含有各種緩衝液成分的液體製劑，於48小時曝光前後之SE-HPLC法的評價結果(聚合物量)圖。 Fig. 7 is a graph showing the results of evaluation by SE-HPLC method (polymer amount) of a liquid preparation containing various buffer components in Example 2 before and after exposure for 48 hours.

[圖8]實施例3進行之將含有各種緩衝液成分的液體製劑，於168小時曝光前後之SE-HPLC法的評價結果(聚合物量)圖。 Fig. 8 is a graph showing the results of evaluation by SE-HPLC method (polymer amount) of a liquid preparation containing various buffer components in Example 3 before and after exposure to 168 hours.

[圖9]實施例4進行將含有各種濃度之作為胺基酸類的精胺酸之液體製劑，於40℃、1個月的保存期間前後之SE-HPLC法的評價結果(聚合物量)圖。 Fig. 9 is a graph showing the results of evaluation of the SE-HPLC method (polymer amount) of a liquid preparation containing arginine as an amino acid at various concentrations and stored at 40 ° C for one month.

[圖10]實施例4進行將含有各種濃度之作為鹽類的氯化鈉之液體製劑，於40℃、1個月的保存期間前後之SE-HPLC法的評價結果(聚合物量)圖。 Fig. 10 is a graph showing the results of evaluation by SE-HPLC method (polymer amount) of a liquid preparation containing sodium chloride as a salt at various concentrations and stored at 40 ° C for one month.

[圖11]實施例4進行將含有各種濃度之作為胺基酸類的甲硫胺酸之液體製劑，於40℃、1個月的保存期間前後之SE-HPLC法的評價結果(聚合物量)圖。 [Fig. 11] Figure 4 shows the results of evaluation of the SE-HPLC method (polymer amount) of a liquid preparation containing methionine as an amino acid at various concentrations at a temperature of 40 ° C for one month. .

[圖12]實施例4進行將含有作為糖或糖醇類的蔗糖或山梨糖醇之液體製劑，於40℃、1個月的保存期間前後之SE-HPLC法的評價結果(聚合物量)圖。 Fig. 12 is a diagram showing the results of evaluation of the SE-HPLC method (polymer amount) of a liquid preparation containing sucrose or sorbitol as a sugar or a sugar alcohol, and a storage period of 40 ° C for one month. .

[圖13]實施例4進行之將含有各種濃度之作為胺基酸類的精胺酸或甲硫胺酸的液體製劑，於48小時曝光前後之SE-HPLC法的評價結果(聚合物量)圖。 Fig. 13 is a graph showing the results of evaluation by SE-HPLC method (polymer amount) of a liquid preparation containing arginine or methionine as an amino acid at various concentrations, which was carried out in Example 4.

[圖14]實施例4進行之將含有各種濃度之氯化鈉液體製劑，以及含有蔗糖或山梨糖醇液體製劑，於48小時曝光前後之SE-HPLC法的評價結果(聚合物量)圖。 Fig. 14 is a graph showing the results of evaluation of the SE-HPLC method (polymer amount) of a liquid preparation containing various concentrations of sodium chloride and a liquid preparation containing sucrose or sorbitol before and after exposure for 48 hours.

[圖15]實施例5進行之使用含有各種濃度之作為非離子性界面活性劑之聚山梨醇酯80液體製劑，振盪試驗實施前後之不溶性微粒子數的測定結果圖。 Fig. 15 is a graph showing the results of measurement of the number of insoluble fine particles before and after the shaking test using a polysorbate 80 liquid preparation containing various concentrations as a nonionic surfactant.

[圖16]實施例5進行之使用含有各種濃度之作為非離子性界面活性劑之聚山梨醇酯80液體製劑，冷凍融解壓力試驗實施前後之不溶性微粒子數的測定結果圖。 Fig. 16 is a graph showing the results of measurement of the number of insoluble fine particles before and after the freeze-thaw pressure test using a polysorbate 80 liquid preparation containing various concentrations as a nonionic surfactant in Example 5.

[圖17]實施例7進行將含有組胺酸、精胺酸及聚山梨醇酯80之各種pH的抗體高濃度液體製劑，於25℃、1個月的保存期間前後之SE-HPLC法的評價結果(聚合物量)圖。 [Fig. 17] Example 7 was carried out by subjecting a high-concentration liquid preparation containing various pHs of histidine acid, arginine acid and polysorbate 80 to SE-HPLC method before and after storage at 25 ° C for one month. Evaluation result (amount of polymer) chart.

[圖18]實施例7進行將含有組胺酸、精胺酸及聚山梨醇酯80之各種pH的抗體高濃度液體製劑，於25℃、1個月的保存期間前後之SE-HPLC法的評價結果(分解物量)圖。 [Fig. 18] Example 7 was carried out by subjecting a high-concentration liquid preparation containing various pHs of histidine acid, arginine acid and polysorbate 80 to SE-HPLC method before and after storage at 25 ° C for one month. Evaluation result (decomposed matter amount) chart.

[圖19]實施例7進行將含有組胺酸、精胺酸及聚山梨醇酯80之各種pH的抗體高濃度液體製劑，於25℃、1個月的保存期間前後之SE-HPLC法的評價結果(不純物量(=聚合物量+分解物量))圖。 [Fig. 19] Example 7 was carried out by subjecting a high-concentration liquid preparation containing various pHs of histidine acid, arginine acid and polysorbate 80 to SE-HPLC method before and after storage at 25 ° C for one month. Evaluation result (amount of impure substance (= amount of polymer + amount of decomposition product)).

[圖20]實施例8進行將含有各種胺基酸類、糖或糖醇類之抗體高濃度液體製劑，於25℃、1個月的保存期間前後之SE-HPLC法的評價結果(聚合物量)圖。 [Fig. 20] Example 8 The evaluation result (polymer amount) of SE-HPLC method before and after the storage period of a high-concentration liquid preparation containing various amino acids, sugars or sugar alcohols at 25 ° C for one month Figure.

[圖21]實施例8進行將含有各種胺基酸類、糖或糖醇類之抗體高濃度液體製劑，於25℃、1個月的保存期間前後之SE-HPLC法的評價結果(分解物量)圖。 [Fig. 21] Example 8 Evaluation results of SE-HPLC method (decomposition amount) of a liquid preparation containing a high concentration of an antibody containing various amino acids, sugars or sugar alcohols at a temperature of 25 ° C for one month Figure.

[圖22]實施例8進行將含有各種胺基酸類、糖或糖醇類之抗體高濃度液體製劑，於25℃、1個月的保存期間前後之SE-HPLC法的評價結果(不純物量(=聚合物量+分解物量))圖。 [Fig. 22] Example 8 The evaluation result of the SE-HPLC method of the high-concentration liquid preparation containing the various amino acids, sugars or sugar alcohols at 25 ° C for one month before storage (the amount of impurities ( = amount of polymer + amount of decomposition product)).

[圖23]實施例9進行之使用含有作為非離子性界面活性劑之聚山梨醇酯80或波洛莎姆188之抗體高濃度液體製劑，振盪試驗實施前後之不溶性微粒子數的測定結果圖。 Fig. 23 is a graph showing the results of measurement of the number of insoluble fine particles before and after the shaking test using the high-concentration liquid preparation containing the polysorbate 80 or the Poloxamer 188 as a nonionic surfactant.

[圖24]實施例9進行之使用含有作為非離子性界面活性劑之聚山梨醇酯80或波洛莎姆188之抗體高濃度液體製劑，冷凍融解壓力試驗實施前後之不溶性微粒子數的測定結果圖。 [Fig. 24] The results of the measurement of the number of insoluble fine particles before and after the freeze-thaw pressure test using the high-concentration liquid preparation containing the polysorbate 80 or the Poloxamer 188 as a nonionic surfactant as carried out in Example 9. Figure.

[圖25]實施例11將含有各種濃度的組胺酸、精胺酸及波洛莎姆188之抗體高濃度液體製劑，於25℃、3個月的保存後之SE-HPLC法及陽離子交換色層管柱分析法(CE-HPLC法)的評價結果，進行統計解析後結果(聚合物的增加量)圖。 [FIG. 25] Example 11 SE-HPLC method and cation exchange of a high-concentration liquid preparation containing various concentrations of histidine acid, arginine acid and Poloxamer 188 at 25 ° C for 3 months storage The results of the evaluation by the color column column analysis method (CE-HPLC method) and the results of the statistical analysis (the amount of increase in the polymer) are shown.

[圖26]實施例11將含有各種濃度的組胺酸、精胺酸及波洛莎姆188之抗體高濃度液體製劑，於25℃、3個月的保存後之SE-HPLC法及CE-HPLC法的評價結果，進行統計解析後結果(分解物的增加量)圖。 [Fig. 26] Example 11 is a high-concentration liquid preparation containing various concentrations of histidine acid, arginine and Poloxamer 188, and stored at 25 ° C for 3 months, SE-HPLC method and CE- As a result of the evaluation by the HPLC method, the result of the statistical analysis (the amount of increase in the decomposition product) is shown.

[圖27]實施例11將含有各種濃度的組胺酸、精胺酸及波洛莎姆188之抗體高濃度液體製劑，於25℃、3個月的保存後之SE-HPLC法及CE-HPLC法的評價結果，進行統計解析後結果(主要波峰的減少量)圖。 [FIG. 27] Example 11 is a high-concentration liquid preparation containing various concentrations of histidine acid, arginine, and Poloxamer 188, and stored at 25 ° C for 3 months, SE-HPLC method and CE- The results of the HPLC method were analyzed by statistical analysis (the amount of reduction of major peaks).

Claims (20)

一種醫藥組成物，其係含有抗人類α9整合素抗體、製藥學容許的緩衝劑，且pH為5.0～7.0之醫藥組成物， 該抗人類α9整合素抗體係選自由下述(1)及(2)所成之群： (1)含有以序列編號1所示胺基酸序列構成之重鏈及以序列編號3所示胺基酸序列構成之輕鏈之抗人類α9整合素抗體，以及， (2)藉由(1)之抗人類α9整合素抗體的轉譯後修飾產生之抗體。A pharmaceutical composition comprising an anti-human α9 integrin antibody, a pharmaceutically acceptable buffer, and a pharmaceutical composition having a pH of 5.0 to 7.0, The anti-human α9 integrin anti-system is selected from the group consisting of the following (1) and (2): (1) an anti-human α9 integrin antibody comprising a heavy chain consisting of the amino acid sequence of SEQ ID NO: 1 and a light chain consisting of the amino acid sequence of SEQ ID NO: 3, and (2) An antibody produced by post-translational modification of the anti-human α9 integrin antibody of (1). 如申請專利範圍第1項之醫藥組成物，其中製藥學容許的緩衝劑係選自由乙酸、檸檬酸、磷酸、組胺酸，以及該等於製藥學容許的鹽所成群之1種或2種以上。The pharmaceutical composition according to claim 1, wherein the pharmaceutically acceptable buffer is selected from the group consisting of acetic acid, citric acid, phosphoric acid, histidine, and one or two of the pharmaceutically acceptable salts. the above. 如申請專利範圍第1項之醫藥組成物，其中製藥學容許的緩衝劑係選自組胺酸或其製藥學容許的鹽。The pharmaceutical composition according to claim 1, wherein the pharmaceutically acceptable buffer is selected from the group consisting of histidine or a pharmaceutically acceptable salt thereof. 如申請專利範圍第1項之醫藥組成物，其中製藥學容許的緩衝劑之濃度係1mmol/L～300mmol/L。The pharmaceutical composition of claim 1, wherein the concentration of the pharmaceutically acceptable buffer is from 1 mmol/L to 300 mmol/L. 如申請專利範圍第1項之醫藥組成物，其係進而含有選自由胺基酸類、鹽類、糖及糖醇類所成群之1種或2種以上。The pharmaceutical composition according to the first aspect of the invention, which further comprises one or more selected from the group consisting of amino acids, salts, sugars and sugar alcohols. 如申請專利範圍第5項之醫藥組成物，其中胺基酸類、鹽類、糖或糖醇類係選自由甲硫胺酸、精胺酸、甘胺酸、離胺酸、鳥胺酸以及該等於製藥學容許的鹽、氯化鈉、蔗糖、山梨糖醇、甘露醇、海藻糖、木糖醇、赤藻糖醇、蘇糖醇、肌醇、半乳糖醇、***糖醇、異麥芽酮糖醇、乳糖醇及麥芽糖醇所成群之1種或2種以上。The pharmaceutical composition according to claim 5, wherein the amino acid, the salt, the sugar or the sugar alcohol is selected from the group consisting of methionine, arginine, glycine, lysine, and auramine; Equal to the pharmaceutically acceptable salt, sodium chloride, sucrose, sorbitol, mannitol, trehalose, xylitol, erythritol, threitol, inositol, galactitol, arabitol, isomalt One or two or more kinds of ketitol, lactitol, and maltitol are grouped. 如申請專利範圍第5項之醫藥組成物，其中胺基酸類、鹽類、糖或糖醇類係精胺酸。The pharmaceutical composition of claim 5, wherein the amino acid, the salt, the sugar or the sugar alcohol is arginine. 如申請專利範圍第5項之醫藥組成物，其中胺基酸類、鹽類、糖或糖醇類之濃度係1mmol/L～400mmol/L。The pharmaceutical composition of claim 5, wherein the concentration of the amino acid, the salt, the sugar or the sugar alcohol is from 1 mmol/L to 400 mmol/L. 如申請專利範圍第1項之醫藥組成物，其係進而含有非離子性界面活性劑。The pharmaceutical composition of claim 1 further comprising a nonionic surfactant. 如申請專利範圍第9項之醫藥組成物，其中非離子性界面活性劑係聚山梨醇酯80及/或波洛莎姆188。The pharmaceutical composition of claim 9, wherein the nonionic surfactant is polysorbate 80 and/or Poloxamm 188. 如申請專利範圍第9項之醫藥組成物，其中非離子性界面活性劑之濃度係0.001％(w/v)～1％(w/v)。The pharmaceutical composition according to claim 9, wherein the concentration of the nonionic surfactant is 0.001% (w/v) to 1% (w/v). 如申請專利範圍第1項之醫藥組成物，其中抗人類α9整合素抗體之濃度係1mg/mL～1000mg/mL。The pharmaceutical composition according to claim 1, wherein the concentration of the anti-human α9 integrin antibody is from 1 mg/mL to 1000 mg/mL. 如申請專利範圍第1項之醫藥組成物，其中醫藥組成物係液體製劑或冷凍乾燥製劑。The pharmaceutical composition of claim 1, wherein the pharmaceutical composition is a liquid preparation or a freeze-dried preparation. 如申請專利範圍第1項之醫藥組成物，其中醫藥組成物係液體製劑。The pharmaceutical composition of claim 1, wherein the pharmaceutical composition is a liquid preparation. 一種醫藥組成物，其係含有抗人類α9整合素抗體、組胺酸或其製藥學容許的鹽，且pH為5.0～7.0之醫藥組成物， 該抗人類α9整合素抗體係選自由下述(1)及(2)所成之群： (1)含有以序列編號1所示胺基酸序列構成之重鏈及以序列編號3所示胺基酸序列構成之輕鏈之抗人類α9整合素抗體，以及， (2)藉由(1)之抗人類α9整合素抗體的轉譯後修飾產生之抗體。A pharmaceutical composition comprising a pharmaceutical composition comprising an anti-human α9 integrin antibody, a histidine acid or a pharmaceutically acceptable salt thereof, and having a pH of 5.0 to 7.0, The anti-human α9 integrin anti-system is selected from the group consisting of the following (1) and (2): (1) an anti-human α9 integrin antibody comprising a heavy chain consisting of the amino acid sequence of SEQ ID NO: 1 and a light chain consisting of the amino acid sequence of SEQ ID NO: 3, and (2) An antibody produced by post-translational modification of the anti-human α9 integrin antibody of (1). 如申請專利範圍第1項之醫藥組成物，其中抗人類α9整合素抗體的轉譯後修飾係重鏈可變區域N端的焦麩胺醯化以及重鏈C端的離胺酸缺失。The pharmaceutical composition of claim 1, wherein the post-translational modification of the anti-human α9 integrin antibody is a glutamate deuteration at the N-terminus of the heavy chain variable region and an acyl acid depletion at the C-terminus of the heavy chain. 一種使用，其係用以製造含有抗人類α9整合素抗體之穩定的醫藥組成物之製藥學容許的緩衝劑、胺基酸類、鹽類、糖或糖醇類的作為光穩定化劑之使用， 該抗人類α9整合素抗體係選自由下述(1)及(2)所成之群： (1)含有以序列編號1所示胺基酸序列構成之重鏈及以序列編號3所示胺基酸序列構成之輕鏈之抗人類α9整合素抗體，以及， (2)藉由(1)之抗人類α9整合素抗體的轉譯後修飾產生之抗體。A use for the manufacture of a pharmaceutically acceptable buffer, amino acid, salt, sugar or sugar alcohol containing a stable pharmaceutical composition of an anti-human α9 integrin antibody, as a photostabilizer, The anti-human α9 integrin anti-system is selected from the group consisting of the following (1) and (2): (1) an anti-human α9 integrin antibody comprising a heavy chain consisting of the amino acid sequence of SEQ ID NO: 1 and a light chain consisting of the amino acid sequence of SEQ ID NO: 3, and (2) An antibody produced by post-translational modification of the anti-human α9 integrin antibody of (1). 如申請專利範圍第17項之使用，其中製藥學容許的緩衝劑係選自由乙酸、檸檬酸、磷酸、組胺酸，以及該等於製藥學容許的鹽所成群之1種或2種以上。The use of the pharmaceutically acceptable buffer is one or more selected from the group consisting of acetic acid, citric acid, phosphoric acid, histidine, and the pharmaceutically acceptable salt. 一種方法，其係於含有抗人類α9整合素抗體之醫藥組成物中，利用製藥學容許的緩衝劑、胺基酸類、鹽類、糖或糖醇類，抑制抗人類α9整合素抗體因曝光而生成分解物或聚合物之方法， 該抗人類α9整合素抗體係選自由下述(1)及(2)所成之群： (1)含有以序列編號1所示胺基酸序列構成之重鏈及以序列編號3所示胺基酸序列構成之輕鏈之抗人類α9整合素抗體，以及， (2)藉由(1)之抗人類α9整合素抗體的轉譯後修飾產生之抗體。A method for inhibiting anti-human α9 integrin antibody by exposure to a pharmaceutical composition containing an anti-human α9 integrin antibody by using a pharmaceutically acceptable buffer, an amino acid, a salt, a sugar or a sugar alcohol a method of producing a decomposition product or a polymer, The anti-human α9 integrin anti-system is selected from the group consisting of the following (1) and (2): (1) an anti-human α9 integrin antibody comprising a heavy chain consisting of the amino acid sequence of SEQ ID NO: 1 and a light chain consisting of the amino acid sequence of SEQ ID NO: 3, and (2) An antibody produced by post-translational modification of the anti-human α9 integrin antibody of (1). 如申請專利範圍第19項之方法，其中製藥學容許的緩衝劑係選自由乙酸、檸檬酸、磷酸、組胺酸，以及該等於製藥學容許的鹽所成群之1種或2種以上。The method of claim 19, wherein the pharmaceutically acceptable buffer is selected from the group consisting of acetic acid, citric acid, phosphoric acid, histidine, and one or more selected from the group consisting of pharmaceutically acceptable salts.