JP2014505474A - Il−17aとil−17fに結合する抗体分子 - Google Patents
Il−17aとil−17fに結合する抗体分子 Download PDFInfo
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Abstract
Description
二価抗体は当分野で知られた方法によって作製され得る(Milstein et al.,1983,Nature 305:537−539;国際公開第93/08829,Traunecker et al.,1991,EMBO J.10:3655−3659)。多価抗体は、多重特異性を含むものであってもよく、単一特異性であってもよい(例えば、国際公開第92/22853号および同第05/113605号参照)。
−フェニルアラニン、チロシンおよびトリプトファン(芳香族側鎖を有するアミノ酸);
−リジン、アルギニンおよびヒスチジン(塩基性側鎖を有するアミノ酸);
−アスパルテートおよびグルタメート(酸性側鎖を有するアミノ酸);
−アスパラギンおよびグルタミン(アミド側鎖を有するアミノ酸);ならびに
−システインおよびメチオニン(含硫側鎖を有するアミノ酸)が挙げられる。同一性および類似性の度合いは、容易に計算することができる(Computational Molecular Biology,Lesk,A.M.,ed.,Oxford University Press,New York,1988;Biocomputing.Informatics and Genome Projects,Smith,D.W.,ed.,Academic Press,New York,1993;Computer Analysis of Sequence Data,Part 1,Griffin,A.M.,and Griffin,H.G.,eds.,Humana Press,New Jersey,1994;Sequence Analysis in Molecular Biology,von Heinje,G.,Academic Press,1987;およびSequence Analysis Primer,Gribskov,M.and Devereux,J.,eds.,M Stockton Press,New York,1991)。
一実施形態において、該病理学的障害は関節リウマチである。
一実施形態において、該病理学的障害はクローン病である。
一実施形態において、該病理学的障害は潰瘍性大腸炎である。
一例では、本発明の抗体は、炎症性疾患または免疫関連疾患の処置において使用される。一例では、炎症性疾患または免疫関連疾患は、関節リウマチ、クローン病および潰瘍性大腸炎からなる群より選択される。
抗体CA028_0496の単離およびヒト化は、以前に国際公開第2008/047134号に報告されている。CA028_0496は、IL−17AとIL−17Fのどちらにも結合し、グラフトされた可変領域gL7とgH9(その配列は国際公開第2008/047134号に示されている)を含むヒト化中和抗体である。重鎖アクセプターフレームワークは、ヒトJH−領域の生殖細胞系統JH4のこの部分に由来するフレームワーク4を有するヒト生殖細胞系統の配列VH3 1−3 3−07である。軽鎖アクセプターフレームワークは、ヒトJK−領域の生殖細胞系統JK1のこの部分に由来するフレームワーク4を有するヒト生殖細胞系統の配列VK1 2−1−(1)L4である。
抗体CA028_00496.g3の親和性成熟型の可変領域の最終配列を図1aと1bに示す。抗体CA028_00496.g3において、重鎖可変領域の配列は、親抗体CA028_00496のものと同じである。対照的に、軽鎖可変領域は5個のアミノ酸が異なる。抗体CA028_00496と抗体CA028_00496.g3の軽鎖間で異なる5つの残基を、図1aにおいて下線で示す。
後述するように、このアッセイ形式は、固定化した抗ヒトIgG Fc−特異的抗体による抗体CA028_00496.g3の捕捉、続いて、捕捉表面上でのヒトIL−17AおよびヒトIL−17Fの滴定とした。
また、IL−17A/Fヘテロ二量体(国際公開第2008/047134号に記載のようにして作製)に対するCA028_0496 g3の親和性も改善され、この場合、親和性は26pMであることがわかった(データ示さず)。
ヒトIL−17Fの中和に対するCA028_00496.g1(国際公開第2008/047134号に既報)およびCA028_00496.g3の効力を、HeLa細胞バイオアッセイを用いて調べた。Hela細胞は、ATCCの細胞バンクから取得した(ATCC CCL−2)。細胞を、10%ウシ胎仔血清、ペニシリン、ゲンタマイシンおよびグルタミンを補給したダルベッコ改変イーグル培地(DMEM)中で培養した。1×104個の細胞を96ウェル平底組織培養プレート内で平板培養した。細胞を一晩インキュベートし、アッセイバッファー中で1回洗浄した。HeLa細胞を、組換えヒトIL−17F(125ng/ml)と腫瘍壊死因子−α(TNF−α)(1ng/ml)の組合せで48時間、種々の濃度の該抗体の存在下で刺激した。HeLa細胞株において、IL−17Fは、TNF−αと相乗作用してIL−6の生成を誘導する。この生成は、特異的MSDアッセイキットを用いて定量され得る。生成した分泌IL−6の量を、Meso Scale Discovery(MSD)アッセイ技術を用いて測定し、IC50値を計算した。HeLa細胞バイオアッセイでの測定時、CA028_00496.g1とCA028_00496.g3では、IL−17Fの生体活性の用量依存性阻害が示された(図7)。このHeLaアッセイでのCA028_00496.g1およびCA028_00496.g3の活性は、IL−17Fの活性の50%を阻害するのに必要とされる用量(IC50)として示した。CA028_00496.g1のIC50は92mg/mLであり、CAO 496.g3では4ng/mLである。
Claims (19)
- ヒトIL−17AおよびヒトIL−17Fに結合し、軽鎖を有する中和抗体であって、該軽鎖の可変ドメインは配列番号:7に示した配列を含むものである中和抗体。
- さらに重鎖を含み、該重鎖の可変ドメインは、CDR−H1が配列番号:1に示した配列、CDR−H2が配列番号:2に示した配列およびCDR−H3が配列番号:3に示した配列を含むものである、請求項1に記載の中和抗体。
- 該重鎖の可変ドメインが配列番号:9に示した配列を含むものである、請求項2に記載の中和抗体。
- IL−17A/IL−17Fヘテロ二量体にも結合する、請求項1〜3のいずれか一項に記載の抗体。
- 完全抗体またはその機能性活性断片である、請求項1〜4のいずれか一項に記載の抗体。
- 該抗体断片が、Fab、Fab’、F(ab’)2、scFvおよびFv断片からなる群より選択される、請求項5に記載の抗体。
- 多重特異性抗体である、請求項1〜4のいずれか一項に記載の抗体。
- ヒトIL−17AおよびヒトIL−17Fに結合し、配列番号:15に示した配列を含む重鎖と配列番号:11に示した配列を含む軽鎖を有する中和抗体。
- 請求項1〜8のいずれか一項に記載の抗体の軽鎖をコードしている単離されたDNA配列。
- 請求項1〜8のいずれか一項に記載の抗体の重鎖と軽鎖をコードしている単離されたDNA配列。
- 請求項9または請求項10に記載のDNA配列を含むクローニングベクターまたは発現ベクター。
- 配列番号:13と配列番号:17に示した配列を含むものである、請求項11に記載のベクター。
- 請求項11または請求項12に記載の1種類以上のクローニングベクターまたは発現ベクターを含む宿主細胞。
- 請求項13に記載の宿主細胞を培養すること、および抗体を単離することを含む、請求項1〜8のいずれか一項に記載の抗体の作製方法。
- 請求項1〜8のいずれか一項に記載の抗体を、薬学的に許容され得る賦形剤、希釈剤または担体のうちの1種類以上との組合せで含む医薬組成物。
- さらに他の活性成分を含む、請求項15に記載の医薬組成物。
- 治療における使用のための、請求項1〜8のいずれか一項に記載の抗体、または請求項15もしくは16に記載の医薬組成物。
- 感染症(ウイルス、細菌、真菌および寄生虫)、感染と関連している内毒素性ショック、関節炎、関節リウマチ、乾癬性関節炎、全身型若年性特発性関節炎(JIA)、全身性エリテマトーデス(SLE)、喘息、慢性閉塞性気道疾患(COAD)、慢性閉塞性肺疾患(COPD)、急性肺障害、骨盤内炎症性疾患、アルツハイマー病、クローン病、炎症性腸疾患、過敏性腸症候群、潰瘍性大腸炎、キャッスルマン病、強直性脊椎炎および他の脊椎関節症、皮膚筋炎、心筋炎、ブドウ膜炎、眼球突出症、自己免疫性甲状腺炎、ペーロニー病、セリアック病、胆嚢疾患、毛巣病、腹膜炎、乾癬、アトピー性皮膚炎、脈管炎、手術による癒着、卒中、自己免疫性糖尿病、I型糖尿病、ライム関節炎、髄膜脳炎、中枢および末梢神経系の免疫媒介性炎症性障害、例えば、多発性硬化症およびギラン・バレー症候群、他の自己免疫障害、膵炎、外傷(手術)、対宿主性移植片病、移植片拒絶、線維形成性障害、例えば、肺線維症、肝線維症、腎線維症、強皮症または全身性硬化症、がん(充実性腫瘍(黒色腫、胚芽腫、肉腫、扁平上皮がん、移行上皮がん、卵巣がんなど)と血液悪性腫瘍、特に、急性骨髄性白血病、慢性骨髄性白血病、慢性リンパ性白血病の両方、胃がんおよび結腸がん)、心臓疾患、例えば、虚血性疾患、例えば、心筋梗塞ならびにアテローム性動脈硬化、血管内凝固、骨吸収、骨粗鬆症、歯周炎および低塩酸症からなる群より選択される病理学的病状の処置または予防における使用のための、請求項1〜8のいずれか一項に記載の抗体、または請求項15もしくは16に記載の医薬組成物。
- IL−17Aおよび/またはIL−17Fによって媒介される、あるいはIL−17Aおよび/またはIL−17Fのレベル増大と関連している病理学的障害の処置または予防のための医薬の製造における、請求項1〜8のいずれか一項に記載の抗体、または請求項15もしくは16に記載の医薬組成物の使用。
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JP2018535961A (ja) * | 2015-10-27 | 2018-12-06 | ユーシービー バイオファルマ エスピーアールエル | 抗il−17a/f抗体を用いた治療方法 |
JP2021119168A (ja) * | 2015-10-27 | 2021-08-12 | ユーシービー バイオファルマ エスアールエル | 抗il−17a/f抗体を用いた治療方法 |
JP2021130707A (ja) * | 2015-10-27 | 2021-09-09 | ユーシービー バイオファルマ エスアールエル | 抗il−17a/f抗体を用いた治療方法 |
JP7176044B2 (ja) | 2015-10-27 | 2022-11-21 | ユーシービー バイオファルマ エスアールエル | 抗il-17a/f抗体を用いた治療方法 |
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