JP2014224120A - 自己免疫疾患の治療のためのBLyS阻害剤および抗CD20剤の組合せ - Google Patents
自己免疫疾患の治療のためのBLyS阻害剤および抗CD20剤の組合せ Download PDFInfo
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Abstract
【解決手段】BLySアンタゴニストが、TACIの細胞外ドメインまたはその機能的断片を含むTACI−Fc融合タンパク質、BAFF−Rの細胞外ドメインまたはその機能的断片を含むBAFF−R−Fc融合タンパク質、あるいはBCMAの細胞外ドメインまたはその機能的断片を含むBCMA−Fc融合タンパク質であり得るFc融合タンパク質である。意図される抗CD20剤のいくつかとしては、リツキサン(登録商標)、オクレリズマブ、オファツムマブ(HuMax−CD20(登録商標))、TRU−015およびDXL625が挙げられるが、CD20と結合する任意の作用物質が適切であり得る。B細胞レベルの低減を必要とする患者、例えば自己免疫疾患に罹患している患者におけるB細胞レベルを低減する。
【選択図】図2
Description
ポリヌクレオチド、ベクター、宿主細胞
ファージ表示
異種ポリペプチドと融合されるかまたは共役されるポリペプチド
ペプチド−ポリマー共役体の構築
検定
FACS分析
製剤処方物
疾患治療
疾患
1.視覚的アナログスケール(VAS)による患者の疼痛査定、
2.疾患活動性(VAS)についての患者の全体的評価、
3.疾患活動性(VAS)についての医者の全体的評価、
4.健康状態質問票により測定される患者の自己評価能力障害、
5.急性期反応物質、CRPまたはESR。
用量投与
投与経路
製品およびキット
BLySアンタゴニストの製造
TACI−Fcの4つのアミノ末端切頭化バージョンが生成される。4つすべてが、配列番号2のアミノ酸残基30と融合されたWO 02/094852に開示されたような修飾されたヒト組織プラスミノーゲン活性因子シグナル配列(配列番号41)を有する。しかしながら、4つのタンパク質は、Fc5が配列番号2のTACIアミノ酸配列と融合された点の位置が異なった。表1は、4つの融合タンパク質の構造を略記する。
チャイニーズハムスター卵巣細胞によるTACI−Fcタンパク質の産生
TACI−Fc発現構築物を用いて、電気穿孔により、動物タンパク質無含有培地中で増殖させた懸濁液適応チャイニーズハムスター卵巣(CHO)DG44細胞をトランスフェクトした(Urlaub et al., Som. Cell. Molec. Genet. 12: 555 (1986))。CHO DG44細胞は、両方のジヒドロフォレートレダクターゼ染色***置での欠失のため、機能性ジヒドロフォレートレダクターゼ遺伝子を欠く。増大された濃度のメトトレキセートの存在下での細胞の増殖は、発現構築物上でのジヒドロフォレートレダクターゼ遺伝子ならびに連結組換えタンパク質コード遺伝子の増幅を生じる。
モノクローナル抗体
検定手順
血液
脾細胞
フローサイトメトリー分析
結果
血液染色
脾細胞
参考文献
カニクイザルにおけるアタシセプトとリツキサンの組合せ
アタシセプトおよび抗CD20モノクローナル抗体(リツキサン(登録商標))同時投与を、カニクイザルにおける31週試験で試験した。
表8:脾臓中の総Bリンパ球(CD40+リンパ球%として)およびメモリーBリンパ球(CD21+/CD27+であるCD40+脾細胞の%)のフローサイトメトリー免疫表現型分類
実施例5
リツキシマブはマウスCD20と結合せず、したがって正常マウスにおいてB細胞を枯渇するために用いられ得ない。さらにまた、in vivoB細胞枯渇のために有効な市販の抗マウスCD20mAbは存在しない。Gong等(J. Immunol 174: 817-826; 2005)は、彼等が抗ヒトCD20mAb(リツキシマブおよびオクレリズマブ)を用いて処置し、これらのmAbで処置されたヒトで観察されるものと同様にB細胞枯渇を誘導するヒトCD20トランスジェニックマウス(細菌人工染色体/BAC技術を用いて生成)の系統を利用した。その論文で、抗CD20mAb療法とBR3−Fc(BAFFR−Ig、BLyS単独阻害薬)とを組合せると、脾臓B細胞の枯渇を有意に増強する、ということも彼等は示した。この実施例は、エール大学のMark Shlomchik(Ahuja et al., J. Immunol 179: 3351-3361; 2007)から認可を得て入手したhCD20 Tgマウスの系統を利用する。
材料および方法
動物の管理、順化および収容
試験終点の要約:
・全血(150 μL;EDTA)を採取し、TおよびB細胞サブセットに関してフローサイトメトリー分析を実施した(表16参照)。
・血清(〜100 μL全血を血清分離試験管中に入れる)を種々の時点で採取して、その後、Luminex検定により総IgG1、IgG2a、IgM、IgEおよびIgAを分析する。
・現時点で、約半数の動物が屠殺されている。残りは、試験の約20週目に屠殺される。
・屠殺時に、脾臓を採取し、フロー分析とその後のIHC/組織学のために加工処理した。採血前に、イソフルラン麻酔下で脾臓摘出を実施した。全脾臓を計量し、記録した後、切片にした。
・屠殺時に、主要末梢リンパ節(鼠径部、腋窩、上腕、頚部および腸間膜)を、考え得る将来的IHCおよび組織学のために採取した。リンパ節を、ホルマリンまたは亜鉛トリス緩衝液で固定し、考え得る将来的使用のために70%アルコール中に保存した。
・組織学的検査用:1/3の脾臓および1つのLLN(左LN)を亜鉛トリス中に入れて、同一組織(右LN)を10%NBF中に採取した。
血清採取:最少で100 μLの全血を血清分離管中に入れて、最低15分間、凝固させた。次に血液を4000 rpmで10分間回転させた。最少で50 μLの血清を、第二容器中に分取した。結成のアリコートを≦60℃で保存した。
a.IHCのための左リンパ節(腸間膜LNを除いて)を含有するカセットを、亜鉛トリス緩衝液中に入れる。
b.組織学のための右リンパ節(腸間膜LNを除いて)を含有するカセットを、10%NBF中に入れる。
c.全腸管(胃から直腸の真上まで)全体を洗浄せずに採取することにより、腸間膜リンパ節を含有するカセットを、10%NBF中に採取する。
試料はすべて、室温で保存した。
2 表現型[B220+/CD19+/huCD20+]は、B220およびネズミCD19表面抗原を、ヒトCD20導入遺伝子を含めて発現するB細胞の集団を説明する。
結果
結論
参考文献
Claims (19)
- 哺乳類におけるB細胞数の低減方法であって、配列番号26の配列を含むポリペプチドを含むBLySアンタゴニストおよび抗CD20剤を投与することを包含する方法。
- BLySアンタゴニストおよび抗CD20剤が相乗的に作用してB細胞レベルを低減する、請求項1記載の方法。
- 低減されるB細胞レベルがメモリーB細胞の数である、請求項2記載の方法。
- 前記抗CD20剤が、リツキサン(登録商標)、オクレリズマブ、オファツムマブ、TRU−015およびDXL625からなる群から選択される、請求項1記載の方法。
- B細胞調節性自己免疫障害の軽減方法であって、治療的有効量の、配列番号26の配列を含むポリペプチドを含むBLySアンタゴニストおよび抗CD20剤を、前記障害に罹患している患者に投与することを包含する方法。
- 前記抗CD20剤が、リツキサン(登録商標)、オクレリズマブ、オファツムマブ、TRU−015およびDXL625からなる群から選択される、請求項5記載の方法。
- 前記抗CD20剤がリツキサン(登録商標)である、請求項6記載の方法。
- BLySアンタゴニストおよび抗CD20剤が相乗的に作用してB細胞レベルを低減する、請求項5記載の方法。
- 低減されるB細胞レベルがメモリーB細胞の数である請求項8記載の方法。
- 前記自己免疫疾患が関節リウマチ、若年性関節リウマチ、全身性紅斑性狼瘡(SLE)、狼瘡性腎炎(LN)、ウェーゲナー病、炎症性腸疾患、特発性血小板減少性紫斑病(ITP)、血栓性血小板減少性紫斑病(TTP)、自己免疫性血小板減少症、多発性硬化症、乾癬、IgA腎症、IgM多発性神経炎、重症筋無力症、血管炎、真性糖尿病、レーノー症候群、シェーグレン症候群および糸球体腎炎からなる群から選択される、請求項5記載の方法。
- 前記自己免疫疾患が全身性紅斑性狼瘡(SLE)である、請求項10記載の方法。
- 前記抗CD20剤がリツキサン(登録商標)である、請求項11記載の方法。
- BLySアンタゴニストが約1〜約25 mg/kgの投与量で投与され、抗CD20が約1〜約25 mg/kgの投与量で投与される、請求項6記載の方法。
- BLySアンタゴニストが約20 mg/kgの投与量で投与され、抗CD20が約20 mg/kgの投与量で投与される、請求項13記載の方法。
- シクロホスファミド(CYC)、アザチオプリン(AZA)、シクロスポリンA(CSA)、ミコフェノール酸モフェチル(MMF)、非ステロイド性抗炎症薬(NSAID)、糖質コルチコイド、プレドニソンおよび疾患修飾性抗リウマチ薬(DMARD)からなる群から選択される免疫抑制薬を用いる療法と併用してBLySアンタゴニストおよび抗CD20剤が投与される、請求項6記載の方法。
- 配列番号26の配列を含むポリペプチドを含むBLySアンタゴニストおよび抗CD20剤を含む、組成物。
- 前記抗CD20剤がリツキサン(登録商標)である、請求項16記載の組成物。
- 配列番号26の配列を含むポリペプチドを含むBLySアンタゴニストおよび抗CD20剤、ならびにラベルを含む製品であって、ここで該ラベルは、該組成物がB細胞調節性自己免疫障害の治療用であることを示す、製品。
- 前記抗CD20剤がリツキサン(登録商標)である、請求項18記載の製品。
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- 2011-03-29 HK HK11103143.7A patent/HK1148948A1/zh unknown
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2014
- 2014-07-03 JP JP2014137551A patent/JP5960756B2/ja active Active
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Patent Citations (2)
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JP2004535182A (ja) * | 2001-05-24 | 2004-11-25 | ザイモジェネティクス,インコーポレイティド | Taci−免疫グロブリン融合タンパク質 |
WO2006068867A1 (en) * | 2004-12-22 | 2006-06-29 | Genentech, Inc. | Combination therapy for b cell disorders |
Non-Patent Citations (1)
Title |
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JPN6015027749; Eur J Clin Pharmacol Vol.63, 20070501, p.647-656 * |
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CA2701329A1 (en) | 2009-04-23 |
EP2233149B1 (en) | 2016-02-10 |
US20090148442A1 (en) | 2009-06-11 |
PL2233149T3 (pl) | 2016-08-31 |
AU2008312406B2 (en) | 2014-03-06 |
JP5960756B2 (ja) | 2016-08-02 |
WO2009052293A1 (en) | 2009-04-23 |
IL204849A (en) | 2017-10-31 |
HK1148948A1 (zh) | 2011-09-23 |
CA2701329C (en) | 2017-08-22 |
AU2008312406A1 (en) | 2009-04-23 |
JP2011500715A (ja) | 2011-01-06 |
ES2565202T3 (es) | 2016-04-01 |
EP2233149A1 (en) | 2010-09-29 |
US8956611B2 (en) | 2015-02-17 |
IL204849A0 (en) | 2010-11-30 |
US20160051671A1 (en) | 2016-02-25 |
HUE030134T2 (en) | 2017-04-28 |
DK2233149T3 (en) | 2016-05-17 |
EP2200631A1 (en) | 2010-06-30 |
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