JP2013523130A - 抗cd40抗体 - Google Patents
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- JP2013523130A JP2013523130A JP2013502780A JP2013502780A JP2013523130A JP 2013523130 A JP2013523130 A JP 2013523130A JP 2013502780 A JP2013502780 A JP 2013502780A JP 2013502780 A JP2013502780 A JP 2013502780A JP 2013523130 A JP2013523130 A JP 2013523130A
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Abstract
Description
CD40は、48kDaのI型内在性膜糖タンパク質であり、そして腫瘍壊死因子(TNF)受容体スーパーファミリーのメンバーである。CD40は、種々の細胞型(正常及び腫瘍性B細胞、指状嵌入細胞、癌腫、上皮細胞(例、ケラチノサイト)、線維芽細胞(例、滑膜細胞)、ならびに血小板を含む)上で発現している。それは、また、単球、マクロファージ、一部の内皮細胞、及び濾胞樹状細胞上に存在する。CD40はB細胞の個体発生において初期に発現され、CD10及びCD19の出現に続いて、しかし、CD21、CD23、CD24の発現、及び表面免疫グロブリンM(sIgM)の出現の前にB細胞前駆体上に出現する(Uckun et al., 1990, Blood 15: 2449)。CD40は、また、扁桃及び骨髄由来の形質細胞上で検出されている(Pellat-Decounynck et al., 1994, Blood 84: 2597)。
本発明はヒト化モノクローナル抗体を提供し、それにおいて、前記抗体は、1nM未満のアンタゴニスト活性IC50を有してヒトCD40に特異的に結合し、かつB細胞増殖において100μg/mLまでアゴニズムを有さず、及び、前記抗体は、さらに、抗体が、非ヒト霊長類におけるインビボでの少なくとも10日間の半減期を有することを特徴とする。
CD40媒介性シグナル伝達は、現在、種々の標的障害に関与するものとして認識されている。これらの障害における介入が治療的に有益でありうることを示す種々の前臨床データの可用性にもかかわらず、自己免疫性疾患の処置において使用することができるアンタゴニスト抗CD40抗体についての必要性が残る。本発明は、好ましい実施態様において、CD40を認識するヒト化抗体に関する。
特定の実施態様において、これらのヒト化抗体の配列は、特定のリードマウス抗体の配列に基づいて同定されている。
抗体のアンタゴニスト特性は、それが1nM未満のB細胞又は樹状細胞のアンタゴニスト活性IC50を有する点で定義される。抗体は、さらに、他の抗CD40抗体(例、抗CD40抗体4D11)と比較して、増加したインビボ半減期を有する優れた薬物動態特性を有する。
この非ヒトアミノ酸配列は、しばしば「インポート」配列として言及され、典型的には、「インポート」抗体ドメイン、特に可変ドメインから取られる。一般的に、ヒト化抗体は、ヒト重鎖又は軽鎖可変ドメインのFRの間に挿入された非ヒト抗体の少なくともCDR又はHVLを含む。本発明は、ヒト生殖系列配列の重鎖及び軽鎖可変ドメインのFRの間に挿入された、表3及び表4に示すマウスモノクローナル抗体に由来するCDRを含む特定のヒト化抗CD40抗体を記載する。特定のマウスFR残基がヒト化抗体の特定の機能に重要でありうるが、従って、ヒト生殖系列配列の重鎖及び軽鎖可変ドメイン残基のいくつかが、対応するマウス配列のものと同じになるように改変されることが理解されるであろう。
本発明の治療用抗体と共役させることができうる化学療法薬剤の多数の例がある。
そのような化学療法薬剤の例は以下を含む:アルキル化薬剤(例えばチオテパ及びシクロホスファミドなど);アルキルスルホネート(例えばブスルファン、インプロスルファン、及びピポスルファなど);アジリジン(例えばベンゾドーパ、カルボコン、メチュアドーパ(meturedopa)、及びウレドパなど);エチレンイミン及びメチルアメラミン(methylamelamine)(アルトレタミン、トリエチレンメラミン、トリエチレンホスホルアミド、トリエチレンチオホスホルアミド、及びトリメチルオロメラミン(trimethylolomelamine)を含む);アセトゲニン(特に、ブラタシン及びブラタシノン);カンプトテシン(合成アナログトポテカンを含む);ブリオスタチン;カリスタチン;CC−1065(そのアドゼレシン、カルゼレシン、及びビゼレシン合成アナログを含む);クリプトフィシン(特に、クリプトフィシン1及びクリプトフィシン8);ドラスタチン、アウリスタチン(アナログモノメチル−アウリスタチンE及びアナログモノメチル−アウリスタチンFを含む);デュオカルマイシン(合成アナログKW−2189及びCBI−TMIを含む);エレウセロビン;パンクラチスタチン;サルコジクチン;スポンジスタチン;ナイトロジェンマスタード(例えばクロランブシル、クロルナファジン、クロロホスファミド、エストラムスチン、イホスファミド、メクロレタミン、酸化塩酸メクロレタミン、メルファラン、ノベムビチン、フェネステリン、プレドニムスチンなど);トロホスファミド、ウラシルマスタード;ニトロソ尿素(例えばカルムスチン、クロロゾトシン、ホテムスチン、ロムスチン、ニムスチン、ラニムスチンなど);抗生物質(例えばエンジイン抗生物質(例、カリケアミシン、特に、カリキアミシンガンマ1I及びカリキアミシンphiI1、例えば、Agnew Chem Intl. Ed. Engl. 33: 183-186(1994)を参照のこと));ダイネミシン(ダイネミシンAを含む);ビスホスホネート(例えばクロドロネートなど);エスペラミシン;ならびにネオカルチノスタチン発色団及び関連する色素タンパク質エンジイン抗生物質色素体)、アクラシノマイシン、アクチノマイシン、オースラマイシン(authramycin)、アザセリン、ブレオマイシン、カクチノマイシン、カラビシン、カルミノマイシン、カルジノフィリン、クロモマイシン、ダクチノマイシン、ダウノルビシン、デトルビシン、6−ジアゾ−5−オキソ−L−ノルロイシン、ドキソルビシン(Adriamycin(商標))(モルホリノ−ドキソルビシン、シアノモルフォリノ−ドキソルビシン、2−ピロリノ−ドキソルビシン、及びデオキシドキソルビシンを含む)、エピルビシン、エソルビシン、イダルビシン、マルセロマイシン、マイトマイシン(例えばマイトマイシンCなど)、ミコフェノール酸、ノガラマイシン、オリボマイシン、ペプロマイシン、ポトフィロマイシン(potfiromycin)、プロマイシン、ケラマイシン(quelamycin)、ロドルビシン、ストレプトニグリン、ストレプトゾシン、ツベルシジン、ウベニメクス、ジノスタチン、ゾルビシン);代謝拮抗剤(例えばメトトレキサート及び5−フルオロウラシル(5−FU)など);葉酸アナログ(例えばデノプテリン、メトトレキサート、プテロプテリン、トリメトレキサートなど);プリンアナログ(例えばフルダラビン、6−メルカプトプリン、チアミプリン、チオグアニンなど);ピリミジンアナログ(例えばアンシタビン、アザシチジン、6−アザウリジン、カルモフール、シタラビン、ジデオキシウリジン、ドキシフルリジン、エノシタビン、フロクスウリジンなど);アンドロゲン(例えばカルステロン、プロピオン酸ドロモスタノロン、エピチオスタノール、メピチオスタン、テストラクトンなど);抗副腎物質(例えばアミノグルテチミド、ミトーテン、トリロスタンなど);葉酸補充薬(例えばフロリン酸;アセグラトン;アルドフォスファミドグリコシド;アミノレブリン酸;エニルウラシル;アムサクリン;ベストラブシル(bestrabucil);ビサントレン;エデトラキサート;デホファミン;デメコルチン;ジアジクオン;エフロルニチン;酢酸エリプチニウム;エポシロン;エトグルシド;硝酸ガリウム;ヒドロキシ尿素;レンチナン;ロニダミン;メイタンシノイド(例えばメイタンシン及びアンサミトシンなど);ミトグアゾン、ミトザントロン;モピダモール;ニトラクリン;ペントスタチン;フェナメット;ピラルビシン;ロソキサントロン;ポドフィリン酸;2−エチルヒドラジド;プロカルバジン;PSK(登録商標);ラゾキサン;リゾキシン(rhizoxin);シゾフィラン;スピロゲルマニウム;テヌアゾン酸;トリアジクオン;2,2’,2’’−トリクロロトリエチルアミン;トリコテセン(特に、T−2トキシン、ベラクリンA、ロリジンA、及びアンギジン);ウレタン;ビンデシン;ダカルバジン;マンノムスチン;ミタブロニトール(mitabronitol);ミトラクトール;ピポブロマン;ガシトシン;アラビノシド(「Ara−C」);シクロフォスファミド;チオテパ;タキソイド、例、パクリタキセル(TAXOL(登録商標)、Bristol-Myers Squibb Oncology,Princeton, NJ)及びドキサタキセル(TAXOTERE(登録商標)、Rhone-Poulenc Rorer,Antony, France);クロラムブシル;ゲムシタビン(Gemzar(商標));6−チオグアニン;メルカプトプリン;メトトレキサート;白金アナログ(例えばシスプラチン及びカルボプラチンなど);ビンブラスチン;白金;エトポシド(VP−16);イホスファミド;ミトザントロン;ビンクリスチン;ビノレルビン(Navelbine(商標));ノバントロン;テニポシド;エダトレキサート;ダウノマイシン;アミノプテリン;キセローダ;イバンドロネート;CPT−11;トポイソメラーゼ阻害剤RFS2000;ジフルオロメチルオルニチン(DMFO);レチノイド(例えばレチノインなど);カペシタビン;ならびに上のいずれかの医薬的に許容可能な塩、酸、又は誘導体。また、この定義において以下が含まれる:腫瘍に対するホルモン作用を調節又は阻害するように作用する抗ホルモン薬剤、例えば抗エストロゲン及び選択的エストロゲン受容体修飾物質(例えば、タモキシフェン(Nolvadex(商標)を含む)、ラロキシフェン、ドロロキシフェン、4−ヒドロキシタモキシフェン、トリオキシフェン、ケオキシフェン、LY117018、オナプリストン、及びトレミフェン(Fareston(商標))を含む);酵素アロマターゼを阻害するアロマターゼ阻害剤、それらは副腎におけるエストロゲン産生を調節する。例えば、4(5) −イミダゾール、アミノグルテチミド、酢酸メゲストロール(Megace(商標))、エキセメスタン、フォルメスタン、ファドロゾール、ボロゾール(Rivisor(商標))、レトロゾール(Femara(商標))、及びアナストロゾール(Arimidex(商標))など;ならびに抗アンドロゲン、例えばフルタミド、ニルタミド、ビカルタミド、ロイプロリド、及びゴセレリン;ならびに上のいずれかの医薬的に許容可能な塩、酸、又は誘導体。これらの薬剤の任意の1つ又は複数を本発明のヒト化抗体に共役させ、種々の障害の処置のための有用な治療用薬剤を提供してもよい。
標識はそれ自体が検出可能でありうる(例、ラジオアイソトープ標識又は蛍光標識)、又は、酵素標識の場合において、検出可能である基質化合物又は組成物の化学変化を触媒しうる。標識したヒト化抗CD40抗体を調製し、種々の適用(インビトロ及びインビボでの診断を含む)において使用することができる。
そのような置換が生物学的活性において変化をもたらす場合、次に、より実質的な変化、表示される「例示的な置換」、又は、アミノ酸分類を参照して以下にさらに記載される通り、導入し、産物をスクリーニングしてもよい。
化合物+DMARDについての6ヵ月でのACR20>50%(GS:オレンシア+DMARD50%対プラセボ+DMARD20%)。さらに他の実施態様において、本発明の組成物は、リツキサンと同様に、関節浸食及び関節腔狭小化について受け入れられているX線スコアリング方法により評価される、1年の期間にわたる構造的損傷の進行を阻害する(52週間後、平均改変Sharpスコア、リツキサン+MTX1.0対プラセボ+MTX2.31)。
− CCFの外観(濁度)
− CCFの濾過特性
− rProteinAの収率
− 溶出及び中和時の濁度
− 可溶性凝集物(SEC)
− 純度/汚染パターン(SDS)
− 電荷パターン(IEF)
表2.2に得られたデータを要約する。データのより詳細な説明は、表2.2に従う。
データは、抗体の曝露と一致した時間点でのCD86発現増加の阻害を示す。
Claims (15)
- 抗体が重鎖及び軽鎖を含み、重鎖配列及び軽鎖配列が以下:
a)配列番号9〜配列番号11からなる群より選択される重鎖CDR1配列、配列番号12〜配列番号15からなる群より選択される重鎖CDR2配列、及び配列番号16〜配列番号17からなる群より選択される重鎖CDR3配列;ならびに
b)軽鎖CDR1配列が配列番号18〜配列番号21からなる群より選択される配列を有し、配列番号22〜配列番号23の軽鎖CDR2配列、及び配列番号24〜配列番号25からなる群より選択される軽鎖CDR3配列
からなる群より選択されるヒト化抗体。 - 抗体が、配列番号10の重鎖CDR1配列、配列番号13の重鎖CDR2配列、及び配列番号16の重鎖CDR3配列を含み;ならびに、抗体が、配列番号19の軽鎖CDR1配列、配列番号22の軽鎖CDR2配列、及び配列番号24の軽鎖CDR3配列を含む、請求項1記載の抗体。
- 抗体が、配列番号9の重鎖CDR1配列、配列番号14の重鎖CDR2配列、及び配列番号16の重鎖CDR3配列を含み;ならびに、抗体が、配列番号20の軽鎖CDR1配列、配列番号22の軽鎖CDR2配列、及び配列番号24の軽鎖CDR3配列を含む、請求項1記載の抗体。
- 抗体が、配列番号11の重鎖CDR1配列、配列番号15の重鎖CDR2配列、及び配列番号17の重鎖CDR3配列を含み;ならびに、抗体が、配列番号21の軽鎖CDR1配列、配列番号23の軽鎖CDR2配列、及び配列番号25の軽鎖CDR3配列を含む、請求項1記載の抗体。
- それぞれ配列番号27及び配列番号26;それぞれ配列番号28及び配列番号26;それぞれ配列番号29及び配列番号26;それぞれ配列番号30及び配列番号26;それぞれ配列番号32及び配列番号31;それぞれ配列番号33及び配列番号31;それぞれ配列番号34及び配列番号31;それぞれ配列番号35及び配列番号31;それぞれ配列番号37及び配列番号36;それぞれ配列番号38及び配列番号36;それぞれ配列番号39及び配列番号36;それぞれ配列番号40及び配列番号36のアミノ酸配列を含む重鎖可変ドメイン及び軽鎖可変領域を有するヒト化モノクローナル抗体又は抗体フラグメント。
- ヒトCD40に特異的に結合する単離抗体又は抗原結合フラグメントであって、以下:
a)配列番号32、配列番号33、配列番号34、又は配列番号35のヒト可変ドメイン重鎖アミノ酸配列のフレームワーク領域のアミノ酸配列と少なくとも90%同一であるアミノ酸配列を有するフレームワーク領域を含むヒト化重鎖可変ドメイン、及び、配列番号31の対応する軽鎖可変と少なくとも90%同一である軽鎖アミノ酸配列を含む;又は
b)配列番号37、配列番号38、配列番号39、又は配列番号40のヒト可変ドメイン重鎖アミノ酸配列のフレームワーク領域のアミノ酸配列と少なくとも90%同一であるアミノ酸配列を有するフレームワーク領域を含むヒト化重鎖可変ドメイン、及び、配列番号36の対応する軽鎖と少なくとも90%同一である軽鎖アミノ酸配列を含む
を含む、単離抗体又は抗原結合フラグメント。 - 抗体又は抗体の抗原結合フラグメントが、1nM未満のアンタゴニスト活性IC50を有し、かつB細胞増殖において100μg/mLまでアゴニズムを有さずにヒトCD40に特異的に結合し、及び、該抗体又は抗体の抗原結合フラグメントは抗体又は抗体の抗原結合フラグメントが、少なくとも10日間の非ヒト霊長類におけるインビボでの半減期を有することをさらに特徴とし;ならびに、該抗体又は抗体の抗原結合フラグメントが、場合により、30mg/kg未満の用量で8日より多いカニクイザルにおける半減期を有する、ヒト化モノクローナル抗体又は抗体の抗原結合フラグメント。
- 哺乳動物に、該哺乳動物においてCD40媒介性の免疫応答を遮断するために十分な量の請求項1〜7のいずれか一項記載の抗体を含む組成物を投与することを含む、哺乳動物においてヒトCD40の機能を遮断する方法。
- 哺乳動物に、哺乳動物において疾患又は障害の症状の1つ又は複数を減少させるために十分な量の請求項1〜7のいずれか一項記載の抗体を含む組成物を投与することを含む、哺乳動物において疾患又は障害を処置又は寛解する方法。
- 疾患又は障害が、移植片対宿主疾患、自己免疫性又は炎症性疾患、及びCD40関連障害からなる群より選択される、請求項9記載の方法。
- 自己免疫性又は炎症性疾患が、関節リウマチ、多発性硬化症、増殖性ループス糸球体腎炎、炎症性腸疾患(IBD)、乾癬、特発性血小板減少性紫斑病(ITP)、クローン病及び全身性エリテマトーデス(SLE)、橋本甲状腺炎、原発性粘液水腫、甲状腺中毒症/グレーブス病、悪性貧血、自己免疫性萎縮性胃炎、自己免疫性心炎、アジソン病、早発閉経、1型糖尿病、グッドパスチャー症候群、重症筋無力症、自己免疫性溶血性貧血、特発性白血球減少症、原発性胆汁性肝硬変、活動性慢性肝炎(HBs Ag陰性)、原因不明肝硬変、シェーグレン症候群、皮膚筋炎、強皮症、混合性結合組織疾患、円板状エリテマトーデス、及び全身性血管炎からなる群より選択される、請求項10記載の方法。
- 選択された第2の治療用薬剤を投与することをさらに含む、請求項8〜11のいずれか一項記載の方法。
- 抗体を、投与の非経口経路、静脈内経路、又は皮下経路により投与する、請求項8〜12のいずれか一項記載の方法。
- 抗体又はその抗原結合フラグメントを、場合により、第2の薬剤に共役させる、(i)請求項1〜7のいずれか一項記載の抗体;及び(ii)医薬的に許容可能な賦形剤を含む医薬的組成物。
- 重鎖可変領域のアミノ酸配列が、配列番号1〜4、配列番号27、配列番号28、配列番号29、配列番号30、配列番号32、配列番号33、配列番号34、配列番号35、配列番号37、配列番号38、配列番号39、配列番号40、配列番号42、配列番号44、配列番号46、配列番号48、配列番号50、配列番号53、配列番号57、配列番号58、配列番号59、配列番号60、配列番号61、配列番号62、配列番号63、配列番号64、配列番号65、配列番号66、配列番号67、配列番号68、配列番号69、配列番号70、配列番号71、配列番号72、又は配列番号73のいずれかを含み;及び、軽鎖可変領域のアミノ酸配列が、配列番号5〜配列番号8、配列番号26、配列番号31、配列番号36、配列番号41、配列番号43、配列番号45、配列番号47、配列番号49、配列番号50、配列番号51、配列番号52、配列番号54、配列番号55、配列番号56、配列番号74、配列番号75、又は配列番号76のいずれかを含む、重鎖可変領域のアミノ酸配列又は軽鎖可変領域をコードする単離ポリヌクレオチド。
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