JPH04187645A - Agent for suppressing action of interleukin 6 - Google Patents
Agent for suppressing action of interleukin 6Info
- Publication number
- JPH04187645A JPH04187645A JP31579290A JP31579290A JPH04187645A JP H04187645 A JPH04187645 A JP H04187645A JP 31579290 A JP31579290 A JP 31579290A JP 31579290 A JP31579290 A JP 31579290A JP H04187645 A JPH04187645 A JP H04187645A
- Authority
- JP
- Japan
- Prior art keywords
- receptor
- antibody
- human
- cell
- mouse
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 108090001005 Interleukin-6 Proteins 0.000 title claims abstract description 34
- 229940100601 interleukin-6 Drugs 0.000 title claims abstract description 29
- 230000009471 action Effects 0.000 title claims abstract description 13
- 102000004889 Interleukin-6 Human genes 0.000 title abstract description 30
- 102000005962 receptors Human genes 0.000 claims abstract description 17
- 108020003175 receptors Proteins 0.000 claims abstract description 17
- 239000003112 inhibitor Substances 0.000 claims description 7
- 208000005485 Thrombocytosis Diseases 0.000 claims 1
- 210000004027 cell Anatomy 0.000 abstract description 10
- 101001076408 Homo sapiens Interleukin-6 Proteins 0.000 abstract description 7
- 102000052611 human IL6 Human genes 0.000 abstract description 7
- 206010035226 Plasma cell myeloma Diseases 0.000 abstract description 6
- 201000000050 myeloid neoplasm Diseases 0.000 abstract description 6
- 230000003053 immunization Effects 0.000 abstract description 5
- 230000002163 immunogen Effects 0.000 abstract description 4
- 230000016784 immunoglobulin production Effects 0.000 abstract description 4
- 201000010099 disease Diseases 0.000 abstract description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 3
- 238000010353 genetic engineering Methods 0.000 abstract description 3
- 208000007766 Kaposi sarcoma Diseases 0.000 abstract description 2
- 241000124008 Mammalia Species 0.000 abstract description 2
- 241001465754 Metazoa Species 0.000 abstract description 2
- 241000283973 Oryctolagus cuniculus Species 0.000 abstract description 2
- 210000004408 hybridoma Anatomy 0.000 abstract description 2
- 210000004881 tumor cell Anatomy 0.000 abstract description 2
- 102000011767 Acute-Phase Proteins Human genes 0.000 abstract 1
- 108010062271 Acute-Phase Proteins Proteins 0.000 abstract 1
- 230000001684 chronic effect Effects 0.000 abstract 1
- 230000001939 inductive effect Effects 0.000 abstract 1
- 230000002062 proliferating effect Effects 0.000 abstract 1
- 230000001737 promoting effect Effects 0.000 abstract 1
- 210000004989 spleen cell Anatomy 0.000 abstract 1
- 102000010781 Interleukin-6 Receptors Human genes 0.000 description 16
- 108010038501 Interleukin-6 Receptors Proteins 0.000 description 13
- 238000000034 method Methods 0.000 description 11
- 241000699670 Mus sp. Species 0.000 description 10
- 230000002401 inhibitory effect Effects 0.000 description 7
- 238000001727 in vivo Methods 0.000 description 6
- 230000000694 effects Effects 0.000 description 5
- 239000012895 dilution Substances 0.000 description 4
- 238000010790 dilution Methods 0.000 description 4
- 101001076414 Mus musculus Interleukin-6 Proteins 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 238000007796 conventional method Methods 0.000 description 3
- 210000002966 serum Anatomy 0.000 description 3
- 208000023275 Autoimmune disease Diseases 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- 241000699666 Mus <mouse, genus> Species 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- 230000002708 enhancing effect Effects 0.000 description 2
- 230000035755 proliferation Effects 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 230000003582 thrombocytopenic effect Effects 0.000 description 2
- NHBKXEKEPDILRR-UHFFFAOYSA-N 2,3-bis(butanoylsulfanyl)propyl butanoate Chemical compound CCCC(=O)OCC(SC(=O)CCC)CSC(=O)CCC NHBKXEKEPDILRR-UHFFFAOYSA-N 0.000 description 1
- 241000700198 Cavia Species 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 241000699660 Mus musculus Species 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 108090000526 Papain Proteins 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 210000000709 aorta Anatomy 0.000 description 1
- VJBCNMFKFZIXHC-UHFFFAOYSA-N azanium;2-(4-methyl-5-oxo-4-propan-2-yl-1h-imidazol-2-yl)quinoline-3-carboxylate Chemical compound N.N1C(=O)C(C(C)C)(C)N=C1C1=NC2=CC=CC=C2C=C1C(O)=O VJBCNMFKFZIXHC-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 230000024245 cell differentiation Effects 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 230000007123 defense Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 239000008121 dextrose Substances 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 230000011132 hemopoiesis Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 238000007912 intraperitoneal administration Methods 0.000 description 1
- 239000007928 intraperitoneal injection Substances 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 239000000644 isotonic solution Substances 0.000 description 1
- 210000005265 lung cell Anatomy 0.000 description 1
- -1 manntol Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 229920000609 methyl cellulose Polymers 0.000 description 1
- 239000001923 methylcellulose Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 210000002569 neuron Anatomy 0.000 description 1
- 238000011580 nude mouse model Methods 0.000 description 1
- 238000012261 overproduction Methods 0.000 description 1
- 229940055729 papain Drugs 0.000 description 1
- 235000019834 papain Nutrition 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 206010039073 rheumatoid arthritis Diseases 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 230000019491 signal transduction Effects 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 208000011580 syndromic disease Diseases 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 206010043554 thrombocytopenia Diseases 0.000 description 1
Abstract
Description
【発明の詳細な説明】
〔産業上の利用分野］
本発明は抗Ｉ Ｌ −６レセプタ一抗体を含んで成る、
生理活性物質であるＴ Ｌ−６の作用抑制剤に関するも
のである。DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The present invention comprises an anti-IL-6 receptor antibody,
This invention relates to an inhibitor of the action of TL-6, which is a physiologically active substance.
〔従来の技術］
インターロイキン−６（ＩＬ−６）は、免疫、造血、炎
症等の生体防御系において重要な役割を果たしており、
生体の増殖分化に広く関与するタンパク譬であるが、一
方ＩＬ−６の生体内での過剰産生は、自己免疫疾患の原
因のひとつとして知られている（岸本、Ｂｌｏｏｄ、
７４． ｐｌ、 １９８９年参照）。したがって生体内
でＩＬ−６の作用を人為的に抑制することは、自己免疫
疾患の新しい治療のメカニズムとして期待されている。[Prior Art] Interleukin-6 (IL-6) plays an important role in biological defense systems such as immunity, hematopoiesis, and inflammation.
IL-6 is a protein that is widely involved in the proliferation and differentiation of living organisms, but on the other hand, overproduction of IL-6 in living organisms is known to be one of the causes of autoimmune diseases (Kishimoto, Blood,
74. pl, 1989). Therefore, artificially suppressing the action of IL-6 in vivo is expected to be a new therapeutic mechanism for autoimmune diseases.
しかしながら、ＩＬ−６の作用を人為的に確実に抑制す
る手段はまだ見出されていない。However, no means for artificially and reliably suppressing the effects of IL-6 has yet been found.
従って本発明はＩＬ−６の作用を抑制するための新規な
手段を提供しようとするものである。Therefore, the present invention seeks to provide a new means for suppressing the effects of IL-6.
〔課題を解決するための手段］
本発明者は前記の課題を解決すべ（種々検討の結果、Ｉ
Ｌ−６レセプターに対する抗体がＩＬ−６の生物学的
作用を抑制するという驚くべき知見を得、これに基いて
本発明を完成した。[Means for Solving the Problems] The present inventor has solved the above problems (as a result of various studies, I
The present invention was completed based on the surprising finding that an antibody against the L-6 receptor suppresses the biological effects of IL-6.
従って本発明は、抗Ｉ Ｌ−６レセプターに対する抗体
を含んで成る、Ｉ Ｌ−６作用抑制剤を提供するもので
ある。Accordingly, the present invention provides an IL-6 action inhibitor comprising an antibody against anti-IL-6 receptor.
（具体的な説明〕
本発明において使用する抗Ｉ Ｌ−６レセブタ一抗体は
、Ｉ Ｌ−６レセプターを特異的ムこ認識するものであ
り、これにはモノクローナル抗体及びポリクローナル抗
体が含まれる。本発明における抗体の製造のために用い
られるヒ）ＴＬ−６レセプターを含む免疫原としては、
ヒトミエローマ細胞Ｕ２６６、で例示されるヒトＩ Ｌ
−６レセプタ一発現細胞、Ｍｌで例示されるヒ） Ｉ
Ｌ−６レセプタ一発現細胞、遺伝子工学的に作製された
可溶性ヒトＩ Ｌ−６レセブクー（特願平１−９７７４
参照）等が挙げられる。(Specific Description) The anti-IL-6 receptor antibody used in the present invention specifically recognizes the IL-6 receptor, and includes monoclonal antibodies and polyclonal antibodies. The immunogen containing the human TL-6 receptor used for the production of antibodies in the invention includes:
Human IL exemplified by human myeloma cells U266.
-6 receptor-expressing cells, exemplified by Ml) I
L-6 receptor-expressing cells, genetically engineered soluble human IL-6 receptor (Japanese Patent Application No. 1-9774)
), etc.
ポリクローナル抗体の製造は、常法に従って、例えば上
記いずれかの免疫原によりマウス、ウサギ、ランド、モ
ルモット等を免疫感作することによって行うことができ
る。Polyclonal antibodies can be produced according to conventional methods, for example, by immunizing mice, rabbits, randos, guinea pigs, etc., with any of the above-mentioned immunogens.
モノクローナル抗体の製造のためのハイプリドーマの作
製も常法に従って行うことができる。例えば上記いずれ
かの免疫原によりマウスやう、ト等の咄乳類を免疫し、
この動物から肺臓細胞を得、これを樹立されたミエロー
マ細胞と融合せしめる。Hybridomas for producing monoclonal antibodies can also be produced according to conventional methods. For example, immunizing mammals such as mice and horses with any of the above immunogens,
Lung cells are obtained from this animal and fused with established myeloma cells.
次に、生体内でのＩ Ｌ−６作用に対する阻害性を有す
るモノクローナル抗体を産生ずるハイブリ１−マをクロ
ーニングする。生体内での１ ＞６作用に対する阻害性
の評価法としては、抗マウスＩ Ｌ−６レセブタ一モノ
クローナル抗体の場合は、本明細書の実施例で擢供され
る方法、すなわちマウスに投与されたヒトｒＬ−６の血
小板増多作用に対する阻害性や、マウスに投与されたヒ
トＩ Ｌ−６の抗体産生増強効果に対する阻害性が挙げ
られる。また抗ヒトＩＬ−６レセプターモノクローナル
抗、体の場合は、ヌードマウスに移植されたＩＬ−６依
存性ヒト癌細胞の増殖に対する阻害性が挙げられる。多
数のクローンをスクリーニングする際は、まずインビト
ロでのＩ Ｌ−６作用に対する阻害性を評価して、一定
数のクローンを選抜しても良い。Next, a hybrida that produces a monoclonal antibody that inhibits IL-6 action in vivo is cloned. In the case of an anti-mouse IL-6 receptor monoclonal antibody, the method for evaluating the inhibitory effect on 1>6 action in vivo is the method provided in the examples of this specification, that is, the method administered to mice. Examples include the inhibitory effect on the thrombocytopenic effect of human rL-6 and the inhibitory effect on the antibody production enhancing effect of human IL-6 administered to mice. In the case of anti-human IL-6 receptor monoclonal antibodies, the inhibitory effect on the proliferation of IL-6-dependent human cancer cells transplanted into nude mice can be mentioned. When screening a large number of clones, a certain number of clones may be selected by first evaluating their inhibitory properties against IL-6 action in vitro.
上記ポリクローナル抗体、及びモノクローナル抗体の回
収・精製方法は、いずれもそれ自体当業界によりよく知
られている方法により行うことができる。The methods for collecting and purifying the polyclonal antibodies and monoclonal antibodies described above can all be performed by methods well known in the art.
本発明中の抗体は、上記方法で調製された天然型の抗体
だけでなく、さらに上記抗体を出発材料として、遺伝子
工学的手法や蛋白化学的方法を用いて作製された人工型
の抗体でもよい。前者の例としては、マウス由来抗ヒト
Ｉ Ｌ −６１／セプター・モノクローナル抗体の可変
領域（Ｖ ｅＴＪ域）とヒト抗体の不変領域（Ｃ？＋１
域）を遺伝子工学的手法で結合させたキメラ型抗体を挙
げることができる。The antibodies of the present invention may be not only natural antibodies prepared by the above method, but also artificial antibodies produced using genetic engineering methods or protein chemical methods using the above antibodies as starting materials. . Examples of the former include the variable region (VeTJ region) of a mouse-derived anti-human IL-61/Scepter monoclonal antibody and the constant region (C?+1) of a human antibody.
One example is a chimeric antibody in which a region) is linked using genetic engineering techniques.
このような抗体の作製方法は、それ自体当業界によりよ
く知られている方法により行うことができる。また後者
の例としては、パパインで抗体分子を切断し調製されう
るＦａｂ領域等が挙げられる。Methods for producing such antibodies can be carried out by methods well known per se in the art. Examples of the latter include Fab regions that can be prepared by cleaving antibody molecules with papain.
なお、ヒトＩ Ｌ−６レセプクーに対するモノクローナ
ル抗体は特願平２−１８９４２０号明細書中に、ＰＭＩ
抗体、及びＭＴ１８抗体として記載されており、これら
を本発明において使用することができる。また、マウス
Ｉ Ｌ−６レセプターに対する抗体は特願平２−２１５
８８６号明細書に記載されたマウスＩ Ｌ−６レセプタ
ーを免疫原として、常法に従って調製したものを本発明
において用いることができる。The monoclonal antibody against human IL-6 receptor is described in Japanese Patent Application No. 189420/1999 as PMI.
antibody, and MT18 antibody, which can be used in the present invention. In addition, antibodies against mouse IL-6 receptor are available from Japanese Patent Application No. 2-215
An immunogen prepared using the mouse IL-6 receptor described in No. 886 according to a conventional method can be used in the present invention.
Ｉ Ｌ−６は細胞膜上のＩＬ−６レセプターと結合し、
さらにｇｐ１３０に会合し、シグナルが伝達される。こ
の様なＩ Ｌ −６シグナル伝達経路の過程において、
抗ＩＬ−６Ｌ−セプクー抗体は、生体内でのＩＬ−６作
用を抑制する効果を有する。本発明の抑制剤により抑制
されるＩ Ｌ−６の作用としては、血小板増多作用、抗
体産生増強作用、泊性朋蛍白誘導作用、腫瘍細胞増殖作
用、神経細胞分化作用等が挙げられる。従って、これら
の作用に起因する疾患、例えばミエローマ、慢性関節リ
ウマチ、カポジ肉腫、キャッスルマン症候群等の治療に
おいて本発明の抑制剤は有効であると考えられる。IL-6 binds to the IL-6 receptor on the cell membrane,
Furthermore, it associates with gp130 and signals are transmitted. In the process of such IL-6 signal transduction pathway,
Anti-IL-6L-Sepcu antibody has the effect of suppressing IL-6 action in vivo. The effects of IL-6 that are inhibited by the inhibitor of the present invention include thrombocytopenia, antibody production enhancement, tomofluorescence induction, tumor cell proliferation, nerve cell differentiation, and the like. Therefore, the inhibitor of the present invention is considered to be effective in treating diseases caused by these effects, such as myeloma, rheumatoid arthritis, Kaposi's sarcoma, and Castleman syndrome.
本発明の抑制剤は、好ましくは非経口投与により、例え
ば静脈内投与、筋肉内投与、経皮投与等により投与する
ことができる。投与量は疾患の種類、患者の状態等によ
り異るが、一般に１０度／ｋｇ〜１０■／ｋｇの範囲で
ある。本発明の抗体は常用の賦形剤、例えば生理食塩水
、ブドウ糖液、マンントール、メチルセルロース、ゼラ
チン等と混合することにより製剤化される。本則はまた
、凍結乾燥品であることができ、これは使用直前に生理
食塩水、５％ブドウ糖液、リンゲル液等の等張液により
再溶解することができる。またつけ加えるならば、ヒト
に対しては、ヒトＩ Ｌ−６レセプターを免疫原として
マウス、ラット等を用いて調製した抗ヒ１−ＩＬ−６レ
セプター抗体をキメラ化した、ヒト人体において異物と
して認識され難いキメラ抗体を用いると艮い。The inhibitor of the present invention can be administered parenterally, for example, intravenously, intramuscularly, transdermally, etc. The dosage varies depending on the type of disease, patient's condition, etc., but is generally in the range of 10°/kg to 10°/kg. The antibodies of the present invention are formulated by mixing with conventional excipients such as physiological saline, glucose solution, manntol, methylcellulose, gelatin, and the like. It can also be a lyophilized product, which can be reconstituted with an isotonic solution such as saline, 5% dextrose, Ringer's solution, etc. immediately before use. In addition, for humans, anti-human IL-6 receptor antibodies prepared using mice, rats, etc. using human IL-6 receptor as an immunogen are chimerized, and are recognized as foreign substances in the human body. It is difficult to use chimeric antibodies that are difficult to detect.
〔実施例］
以下本発明をさらに詳細に説明するために実施例を記載
するが、本発明はこれら実施例により限定されるもので
はない。[Examples] Examples will be described below to explain the present invention in more detail, but the present invention is not limited by these Examples.
夫ＩＬＬ、 マウスＪ Ｌ−６レセプター・ポリク抗マ
ウスＩ Ｌ−６レセプター・ポリクローナル抗体は、Ｃ
ＨＯ細胞由来可溶性マウスＩＬ−６レセプター（特願平
１−２９２２３０号、特願平２−２１５８８６号参照）
をモルモフトに免疫して調製した。ＩＣＲマウス（ａ性
、５週齢〕１群５匹、全４群を用い、その内の一群を未
投与群（対照）とし、他の３群をそれぞれ、■９０μｇ
の大腸菌由来リコンビナンＮＩＬ−６を１日１回腹腔打
ちに投与する群、０１日１回、９００硝の抗マウスＴＬ
−６レセプター・ポリクローナル抗体を腹腔内に投与し
た後、９０■の前記リコンビナン）ＩＬ−６を投与する
群、■９００■の抗マウスＩＬ−６レセプター・ポリク
ローナル抗体と９０ｔｔｇのリコンビナントＩＬ−６を
混合して１日１回投与する群とし、それぞれ５日間連続
投与した。６日目に後大動脈より全採血し、血小板数を
ヘモサイトメーターＰＣ−６０１（エルマ社製）で計数
した。この結果を第１Ｖに示す。Husband ILL, mouse J L-6 receptor polyclonal antibody, anti-mouse IL-6 receptor polyclonal antibody, C
Soluble mouse IL-6 receptor derived from HO cells (see Japanese Patent Application No. 1-292230, Japanese Patent Application No. 2-215886)
was prepared by immunizing Mormoft. ICR mice (sex A, 5 weeks old): 5 mice per group, 4 groups in total; one group was treated as an untreated group (control), and the other 3 groups received 90 μg each.
A group in which recombinant NIL-6 derived from E. coli was administered by intraperitoneal injection once a day, 900 N anti-mouse TL once a day.
-6 receptor polyclonal antibody was administered intraperitoneally, and then 90■ of the above recombinant IL-6 was administered, and 900■ of anti-mouse IL-6 receptor polyclonal antibody and 90ttg of recombinant IL-6 were mixed. Each group was administered once a day for 5 consecutive days. On the 6th day, all blood was collected from the posterior aorta, and the number of platelets was counted using a hemocytometer PC-601 (manufactured by Elma). The results are shown in Section 1V.
第２図から明らかなように、９０鱈／日のＩＬ−６を投
与した群では有意に血小板数が増加しているのに対し、
９００ｉ／日の抗体を投与した群（前記■、■の群）で
は、いずれもＩ Ｌ −６を投与したにもかかわらずＩ
Ｌ−６未投与群（対照）と同しレベル又はそれ以外で
あった。このことから、抗マウスＩＬ−６レセプター・
ポリクローナル抗体は、ｉｎ ｖｉｖｏでのＩ Ｌ−６
の血小板増多効果に対する抑制効果を示すことが観察さ
れた。As is clear from Figure 2, the number of platelets increased significantly in the group receiving IL-6 at 90 cod/day;
In the groups to which 900 i/day of antibody was administered (groups ■ and ■ above), despite the administration of IL-6, I
It was at the same level as the L-6 non-administered group (control) or other than that. From this, anti-mouse IL-6 receptor
Polyclonal antibodies can be used to detect IL-6 in vivo.
was observed to exhibit an inhibitory effect on the thrombocytopenic effect of .
ＢＡＬＢ／Ｃ７ウス（雄性、５週齢）に、１ ｍｇのＤ
ＮＰ／ＢＳＡ （シグマ社）を皮下投与し、翌日から１
日１ 回０〜１０ｉ（７） Ｉ Ｌ −６及びＯ〜１０
０ｘの抗マウスＩＬ−６レセプクー・ポリクローナル抗
体（実施例１参照）の混合液を腹腔内に６日間連続投与
した。７日目に全採血を行い、血清分離をし、抗ＤＮＰ
／ＢＳＡ抗体価を通常のＥＬＩＳＡ法で測定した。BALB/C7 mice (male, 5 weeks old) were given 1 mg of D.
NP/BSA (Sigma) was administered subcutaneously, and from the next day
Once a day 0-10i (7) I L -6 and O-10
A mixture of 0x anti-mouse IL-6 receptor polyclonal antibody (see Example 1) was administered intraperitoneally for 6 consecutive days. On the 7th day, whole blood was collected, serum was separated, and anti-DNP
/BSA antibody titer was measured by a conventional ELISA method.
即ち各血清に対し１０倍稀釈を最高濃度とし、さらに２
倍稀釈列を作り、これらの抗ＤＮＰ／ＢＳＡに特異的な
ＩｇＧ量を測定した。That is, for each serum, a 10-fold dilution is the highest concentration, and a further 2
A double dilution series was prepared and the amount of these anti-DNP/BSA-specific IgGs was measured.
第２図は横軸にその稀釈倍数を、継軸にＩｇＧ量に相関
した４０５硝ｍでの吸光度を示したもので、この図から
明らかなように、１０μｇ／日のＩ Ｌ−６投与により
増加した抗ＤＮＰ／ＢＳＡ抗体価は、投与した抗体量に
依存して下がり、特に１００μｇ／日の抗体投与では、
ＩＬ−６未投与のレベル以下に下がった。このように、
抗マウスＩ Ｌ−６レセプター・ポリクローナル抗体の
、ｉｎ ｖｉｖｏでのＩＬ−６の抗体産生増強効果に対
する抑制効果が観察された。In Figure 2, the horizontal axis shows the dilution factor, and the joint axis shows the absorbance at 405 Nm, which is correlated with the IgG amount.As is clear from this figure, administration of 10 μg/day of IL-6 The increased anti-DNP/BSA antibody titer decreased depending on the amount of antibody administered, and especially when the antibody was administered at 100 μg/day,
It decreased to below the level when no IL-6 was administered. in this way,
The inhibitory effect of the anti-mouse IL-6 receptor polyclonal antibody on the in vivo antibody production enhancing effect of IL-6 was observed.
第１図は、Ｉ Ｌ−６及び抗マウスＩＬ−６レセブター
・ポリクローナル抗体を投与されたＩＣＲマウスの血小
板数を示す。
第２圀は、ＩＬ−６及び抗マウスＩＬ−６レセプター・
ポリクローナル抗体を投与されたＤＮＰ／ＢＳＡ感作マ
ウスの抗ＤＮＰ／ＢＳＡ抗体価を示す。
（ｘｉ○ム／μＬ）
群
＃１：サンプル未投与群
＃２：ＩＬ−６９０μｇ腹腔投与群
＃３：抗マウスＩＬ−６レセプクー・ポリクローナル抗
体９００／７ｇ静脈投与ＩＬ−６９０μｇａ腔投与群
○Ｄ４０５
血清希釈倍率FIG. 1 shows platelet counts in ICR mice administered IL-6 and anti-mouse IL-6 receptor polyclonal antibody. The second area is IL-6 and anti-mouse IL-6 receptor.
The anti-DNP/BSA antibody titers of DNP/BSA-sensitized mice administered with polyclonal antibodies are shown. (xi○mu/μL) Group #1: Sample not administered Group #2: IL-690μg intraperitoneal administration group #3: Anti-mouse IL-6 receptor polyclonal antibody 900/7g intravenous administration IL-690μga intracavitary administration group○D405 Serum Dilution ratio
Claims (1)
体を含んで成るＩＬ−６作用抑制剤。 ２、ＩＬ−６による血小板増多作用を抑制する、請求項
１に記載のＩＬ−６作用抑制剤。[Claims] 1. An IL-6 action inhibitor comprising an anti-interleukin-6 (IL-6) receptor antibody. 2. The IL-6 action inhibitor according to claim 1, which inhibits the thrombocytosis action caused by IL-6.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP31579290A JPH04187645A (en) | 1990-11-22 | 1990-11-22 | Agent for suppressing action of interleukin 6 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP31579290A JPH04187645A (en) | 1990-11-22 | 1990-11-22 | Agent for suppressing action of interleukin 6 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH04187645A true JPH04187645A (en) | 1992-07-06 |
Family
ID=18069612
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP31579290A Pending JPH04187645A (en) | 1990-11-22 | 1990-11-22 | Agent for suppressing action of interleukin 6 |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH04187645A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1996025174A1 (en) * | 1995-02-13 | 1996-08-22 | Chugai Seiyaku Kabushiki Kaisha | Muscle protein decomposition inhibitor containing il-6 receptor antibody |
| EP0783893A1 (en) | 1994-10-07 | 1997-07-16 | Chugai Seiyaku Kabushiki Kaisha | Rheumatoid arthritis remedy containing il-6 antagonist as active ingredient |
| CN1306963C (en) * | 1994-10-21 | 2007-03-28 | 岸本忠三 | Remedy for diseases caused by IL-6 production |
| US8017121B2 (en) | 1994-06-30 | 2011-09-13 | Chugai Seiyaku Kabushika Kaisha | Chronic rheumatoid arthritis therapy containing IL-6 antagonist as effective component |
-
1990
- 1990-11-22 JP JP31579290A patent/JPH04187645A/en active Pending
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8017121B2 (en) | 1994-06-30 | 2011-09-13 | Chugai Seiyaku Kabushika Kaisha | Chronic rheumatoid arthritis therapy containing IL-6 antagonist as effective component |
| EP0783893A1 (en) | 1994-10-07 | 1997-07-16 | Chugai Seiyaku Kabushiki Kaisha | Rheumatoid arthritis remedy containing il-6 antagonist as active ingredient |
| EP2077120A3 (en) * | 1994-10-07 | 2009-07-15 | Chugai Seiyaku Kabushiki Kaisha | Rheumatoid arthritis remedy containing il-6 antagonist as active ingredient |
| EP2107070A1 (en) * | 1994-10-07 | 2009-10-07 | Chugai Seiyaku Kabushiki Kaisha | Rheumatoid arthritis remedy containing IL-6 antagonist as active ingredient |
| CN1306963C (en) * | 1994-10-21 | 2007-03-28 | 岸本忠三 | Remedy for diseases caused by IL-6 production |
| WO1996025174A1 (en) * | 1995-02-13 | 1996-08-22 | Chugai Seiyaku Kabushiki Kaisha | Muscle protein decomposition inhibitor containing il-6 receptor antibody |
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