JP5016487B2 - 新規な抗igf−ir抗体及びその使用 - Google Patents
新規な抗igf−ir抗体及びその使用 Download PDFInfo
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- JP5016487B2 JP5016487B2 JP2007523178A JP2007523178A JP5016487B2 JP 5016487 B2 JP5016487 B2 JP 5016487B2 JP 2007523178 A JP2007523178 A JP 2007523178A JP 2007523178 A JP2007523178 A JP 2007523178A JP 5016487 B2 JP5016487 B2 JP 5016487B2
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
- C07K16/2896—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against molecules with a "CD"-designation, not provided for elsewhere
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P13/00—Drugs for disorders of the urinary system
- A61P13/08—Drugs for disorders of the urinary system of the prostate
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P15/00—Drugs for genital or sexual disorders; Contraceptives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/06—Antipsoriatics
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
Description
a)本発明による抗体又はその機能的断片の一つをコードする核酸、DNA又はRNA、及び
b)a)で定義されたような核酸に相補的な核酸
から選択されることを特徴とする、単離された核酸に関する。
a)本発明による宿主細胞を培地内で適切な培養条件で培養する工程、及び
b)このようにして生成された上記抗体又はその機能的断片の一つを、培養培地又は上記培養した細胞から回収する段階。
a)生体試料を、本発明による抗体又はその機能的断片の一つと接触させる工程、及び
b)形成された可能性のあるIGF−IR/抗体複合体を明らかにする工程
を含むことを特徴とする。
(a)本発明による抗体又はその機能的断片の一つ、
(b)所望により、免疫学的反応に適した培地を形成するための試薬、
(c)所望により、免疫学的反応によって生成したIGF−IR/抗体複合体を明らかにすることが可能な試薬
を含むことを特徴とする。
−図1は、MCF−7モデルにおける抗IGF−1R抗体のin vitroでの評価を示す図である。
−図2及び3は、DU145での抗IGF−1R抗体のin vivoでの評価を示す図である。
−図4は、IGF−IRを発現する無傷細胞での[125I]−IGF−1の排除を示す図である。
−図5は、イムノキャプチャーしたHR−Aでの[125I]−IGF−1の排除を示す図である。
−図6は、イムノキャプチャーしたHR−Bでの[125I]−IGF−1の排除を示す図である。
−図7は、IR−Aを発現する無傷細胞での[125I]−INSの排除を示す図である。
−図8は、IR−Bを発現する無傷細胞での[125I]−INSの排除を示す図である。
可溶性のα2−β2ヘテロ四量体組換えヒトIGF−1R(R&Dシステム、米国ミネアポリス)で免疫されたBALB/cマウス由来の脾細胞とSP2/0−Ag14骨髄腫細胞株との融合によってハイブリドーマを作製した。得られたマウス抗体は、まずMCF−7細胞でのELISA及びFACS分析によってスクリーニングした。その後、Sf9−IGF−1R細胞とSf9−IR細胞で最後のスクリーニングを行い、IGF−1及びIRの両方を認識する抗体を除外した。選択されたMAb(ELISAで陽性であり、MCF−7細胞の野生型受容体を認識する)を、腹水として産生し、プロテインAクロマトグラフィで精製した後、in vitro及び/又はin vivoで試験し、結果を表1にまとめた。
方法
ATCCから入手したMCF−7細胞を、フェノールレッドフリーRPMI培地(インビトロジェン社、英国、スコットランド)、10%FCS(インビトロジェン社)、1%L−グルタミン(インビトロジェン社)中で常法により培養した。MCF−7細胞を、96ウェル組織培養プレートの無血清培地中に5×104細胞/ウェルの密度で播種した。24時間後、最終濃度が5μg/mlの各試験抗体の非存在下又は存在下で、IGF1を1〜50ng/mlの範囲の投与量で培地に添加した。3日後、0.5μCiの[3H]チミジン(アマシャムバイオサイエンス AB、スウェーデン国、ウプサラ)を細胞に16時間パルスした。トリクロロ酢酸に不溶性のDNAに組み込まれた[3H]チミジンの量を、液体シンチレーション計数によって定量した。結果を増殖指数(細胞+IGF1+抗体のcpm/細胞+抗体のみのcpm)として表す。
in vitroでの評価は、細胞分裂促進活性に関するMabの第一のスクリーニングであった。これらの分析は、腹水として産生された、作製された抗体を、IGF1と同時にMCF−7細胞に添加し、市販のαIR3Mabと比較して、その抗体と少なくとも同等に有効である抗体を選択することによって行った。先の結果の表2*中で(+)と記載されている陽性のMab(5つのMab)は、最も高い投与量のIGF1(50ng/ml)で細胞を刺激したときに増殖指数<5を示すものである。図1は、6つの強いin vitro阻害剤のうちの4つ(2D10、12D5、12B1、13F5)のin vitro活性を示す。2F2及び21E3のMabは、(±)*Mab(最も高いIGF1濃度の場合に5<増殖指数<15)であると考えられ、7G3及び2B10は、非中和抗体(増殖指数>15)であると考えられた。21E3がIgG2アイソタイプのMabにすぎないことは興味深い。
方法
ATCCから入手したDU145細胞を、DMEM培地(インビトロジェン社、英国、スコットランド)、10%FCS(インビトロジェン社)、1%L−グルタミン(インビトロジェン社)中で常法により培養した。細胞を移植の二日前に分割して、対数増殖期となるようにした。PBS中の200万個のDU145細胞をSwissヌードマウスに移植した。移植の一日後、マウスを6頭の群に分割した。1週間に3回、200μgの各試験抗体で、腫瘍と反対の位置においてマウスを皮下注射処理した。コントロール群は、最初のスクリーニングにおいてマウスのアイソタイプのコントロール(EC2)で処理するか、又は続くスクリーニングにおいてPBSで処理するとしたが、これは、最初の実験においてマウスのこれら2つの群の間で腫瘍成長の差異が観察されなかったことが示されたためである。腫瘍の体積を一週間に一度測定し、次の式によって算出した:π/6×長さ×幅×高さ。
三つのin vivoでの実験を、Mabパネルを試験するために行った。図2及び図3は、13F5、2D10及び6E5が、DU145細胞のin vivoでの成長を有意に阻害することを示す。統計分析(マンホイットニー検定)の結果を表2に示す。
この研究に用いた細胞を以下に列挙する。
・R+:IGF−I受容体(IGF−IR)のcDNAを安定的にトランスフェクトしたR−繊維芽細胞、
・R−/IR−A:インスリン受容体アイソフォームA(IR−A)のcDNAを安定的にトランスフェクトしたR−繊維芽細胞、
・R−/IR−B:インスリン受容体アイソフォームB(IR−B)のcDNAを安定的にトランスフェクトしたR−繊維芽細胞、
・R+/IR−A:IGF−I及びインスリン受容体アイソフォームAのcDNAを安定的にコトランスフェクトし、それによりハイブリッド受容体A(ハイブリッド−RsA)を発現するR−繊維芽細胞、
・R+/IR−B:IGF−I及びインスリン受容体アイソフォームBのcDNAを安定的にコトランスフェクトし、それによりハイブリッド受容体A(ハイブリッド−RsB)を発現するR−繊維芽細胞。
Claims (1)
- ヒトインスリン様成長因子I受容体IGF−IRに結合することができる、単離された抗体又はその機能的断片の一つであって、配列番号1、2及び3の配列を有する3つの相補性決定領域CDRを含む軽鎖を含み、かつ、配列番号4、5及び6の配列を有する3つのCDRを含む重鎖を含むことを特徴とする、単離された抗体又はその機能的断片の一つ。
Applications Claiming Priority (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US59193204P | 2004-07-29 | 2004-07-29 | |
US60/591,932 | 2004-07-29 | ||
FR0408379 | 2004-07-29 | ||
FR0408379A FR2873699B1 (fr) | 2004-07-29 | 2004-07-29 | Nouveaux anticorps anti igf ir rt leurs utilisations |
PCT/IB2005/002619 WO2006013472A2 (en) | 2004-07-29 | 2005-07-27 | Novel anti-igf-ir antibodies and uses thereof |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
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JP2012038645A Division JP2012153692A (ja) | 2004-07-29 | 2012-02-24 | 新規な抗igf−ir抗体及びその使用 |
Publications (2)
Publication Number | Publication Date |
---|---|
JP2008515773A JP2008515773A (ja) | 2008-05-15 |
JP5016487B2 true JP5016487B2 (ja) | 2012-09-05 |
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JP2007523178A Expired - Fee Related JP5016487B2 (ja) | 2004-07-29 | 2005-07-27 | 新規な抗igf−ir抗体及びその使用 |
JP2012038645A Pending JP2012153692A (ja) | 2004-07-29 | 2012-02-24 | 新規な抗igf−ir抗体及びその使用 |
JP2014053009A Pending JP2014193862A (ja) | 2004-07-29 | 2014-03-17 | 新規な抗igf−ir抗体及びその使用 |
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JP2012038645A Pending JP2012153692A (ja) | 2004-07-29 | 2012-02-24 | 新規な抗igf−ir抗体及びその使用 |
JP2014053009A Pending JP2014193862A (ja) | 2004-07-29 | 2014-03-17 | 新規な抗igf−ir抗体及びその使用 |
Country Status (23)
Country | Link |
---|---|
US (3) | US7854930B2 (ja) |
EP (4) | EP2365006A3 (ja) |
JP (3) | JP5016487B2 (ja) |
KR (1) | KR101220635B1 (ja) |
CN (2) | CN103951752A (ja) |
AR (1) | AR050036A1 (ja) |
AU (1) | AU2005268505B2 (ja) |
BR (1) | BRPI0513890A (ja) |
CA (1) | CA2575348A1 (ja) |
CO (1) | CO5700152A1 (ja) |
FR (1) | FR2873699B1 (ja) |
GT (1) | GT200500205A (ja) |
HK (1) | HK1102600A1 (ja) |
IL (2) | IL181025A (ja) |
MX (1) | MX2007001109A (ja) |
NO (1) | NO20071093L (ja) |
NZ (2) | NZ569718A (ja) |
PA (1) | PA8640601A1 (ja) |
RU (2) | RU2406729C2 (ja) |
SV (1) | SV2006002182A (ja) |
TW (1) | TWI432451B (ja) |
WO (1) | WO2006013472A2 (ja) |
ZA (1) | ZA200700593B (ja) |
Cited By (1)
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JP2012153692A (ja) * | 2004-07-29 | 2012-08-16 | Pierre Fabre Medicament | 新規な抗igf−ir抗体及びその使用 |
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