JP2016074740A - 単球免疫応答を誘発するための、組換えlag−3タンパク質またはその誘導体の使用 - Google Patents
単球免疫応答を誘発するための、組換えlag−3タンパク質またはその誘導体の使用 Download PDFInfo
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Abstract
【解決手段】本発明は、単球媒介性免疫応答を増加させるための、特に、血中の単球数の増加を引き起こすための、組換えLAG−3タンパク質またはその誘導体の使用に関する。これによって、感染症治療または癌治療のための新しい治療薬の開発に用いることができる。本発明者らは、ヒトLAG−3またはその誘導体を、転移性乳癌(MBC)または転移性腎細胞癌(MRCC)などの極めて悪性の腫瘍を有する患者に接種したところ、ヒトLAG−3またはその誘導体が単球依存性の強力な免疫力を生じさせるという結果を、全く予想外に、見出した。
【選択図】なし
Description
本発明は、単球媒介免疫応答を促進するための、組換えLAG−3またはその誘導体の使用に関する。
活性化ヒトCD4+およびCD8+T細胞および活性化NK細胞において発現しているリンパ球活性化遺伝子3(hlag−3)は、4つの細胞外免疫グロブリンスーパーファミリー(IgSF)ドメインを有する、503アミノ酸のI型膜タンパク質(LAG−3)をコードしている[1]。マウスリンパ球活性化遺伝子3(mlag−3)がクローン化され、hlag−3との間に約70%の類似性が確認され、細胞質内テールに、同じプロリンリッチモチーフが見られた。
、TRIEBELet. al.[3]、およびHUARD et. al.[4]において、LAG−3のさらなる
情報および免疫賦活剤としての使用が開示されている。
・IgGのFc断片の受容体(CD16、CD32、CD64)、
・IgEのFc断片の受容体(CD23)、
・補体断片の受容体(CD11b、CD21/CD35)、
・白血球付着タンパク質(CD11a、CD11c)、
・グラム−菌のLPS結合促進タンパク質(CD14)、
・チロシンフォスタファーゼ活性を有するタンパク質(CD45)。
本発明者らは、ヒトLAG−3またはその誘導体を、転移性乳癌(MBC)または転移性腎細胞癌(MRCC)などの極めて悪性の腫瘍を有する患者に接種したところ、ヒトLAG−3またはその誘導体が単球依存性の強力な免疫力を生じさせるという結果を、全く予想外に、見出した。
・全LAG−3タンパク質、
・LAG−3の可溶性ポリペプチド断片であり、4つの免疫グロブリン細胞外ドメインのうち少なくとも1つを有するLAG−3の可溶性ポリペプチド断片。すなわち、フランス特許出願番号FR9000126に開示されているLAG−3配列の23番目のアミノ酸から448番目のアミノ酸までの細胞外領域を含んでいるLAG−3の可溶性部分、
・第1および第2のドメインの実質的に全てからなるLAG−3断片;
・第1および第2のドメインの実質的に全てもしくは4つのドメインの全てからなるLAG−3断片(例えば、国際公開WO95/30750)、
・D1およびD2細胞外ドメインを含み、以下の置換からなる可溶性LAG−3の変異体またはその断片:
*以下のいずれかの位置におけるアミノ酸の置換:
ARGがGLUに置換されている73位のアミノ酸、
ARGがALAまたはGLUに置換されている75位のアミノ酸、
ARGがGLUに置換されている76位のアミノ酸、
もしくはこれらのうち2つ以上の組合せによるアミノ酸の置換、
*以下のいずれかの位置におけるアミノ酸の置換:
ASPがALAに置換されている30位のアミノ酸、
HISがALAに置換されている56位のアミノ酸、
TYFがPHEに置換されている77位のアミノ酸、
ARGがALAに置換されている88位のアミノ酸、
ARGがALAに置換されている103位のアミノ酸、
ASPがGLUに置換されている109位のアミノ酸、
ARGがALAに置換されている115位のアミノ酸、
もしくは54位のアミノ酸と66位のアミノ酸の間の領域の欠失、
もしくはこれらのうち2つ以上の組合せによるアミノ酸の置換。
・D1、D2、D3を含有する可溶性52kDaタンパク質を含んでなるLAG−3の生理学的変種。
・ヒトIgG1 Fcに融合したhLAG−3の細胞外ドメインをコードするプラスミドを形質移入したチャイニーズハムスター卵巣細胞において生成される200kDA二量体である、組換え可溶性ヒトLAG−3lg融合タンパク質(IMP321)。
例えば、本願発明は以下の項目を提供する。
(項目1)
単球数の増加を促す感染症治療用または癌治療用の医薬を製造するための、単球媒介免疫応答を引き起こす組換えLAG−3タンパク質またはその誘導体の使用。
(項目2)
上記医薬が、組換えLAG−3タンパク質またはその誘導体の効果的な複数回投与単位からなることを特徴とする項目1に記載の使用。
(項目3)
組換えLAG−3タンパク質またはその誘導体の上記複数回投与単位が、少なくとも12週間、好ましくは少なくとも24週間にわたって1週間〜数週間ごとに1回、13日±2日の無投与期間をあけて1回の投与単位を投与できるように調剤されていることを特徴とする項目2に記載の使用。
(項目4)
組換えLAG−3タンパク質またはその誘導体の上記1回の投与単位が、投与を必要としている肥満度指数(体重/身長 2 )が18〜30kg/m 2 の患者に対して、0.25〜30mg、好ましくは6〜30mgの範囲で、組換えLAG−3タンパク質またはその誘導体を皮下投与または静脈投与できるように調剤されていることを特徴とする項目2
または3に記載の使用。
(項目5)
組換えLAG−3タンパク質またはその誘導体の上記投与単位が、抗癌もしくは抗感染症の免疫治療特性または化学治療特性を有する化合物とともに投与できるように調剤されていることを特徴とする項目2〜4のいずれか1項に記載の使用。
(項目6)
組換えLAG−3タンパク質またはその誘導体の1回の投与単位が、0.25〜30mg、好ましくは6〜30mgの組換えLAG−3タンパク質またはその誘導体を含んでなることを特徴とする項目5に記載の使用。
(項目7)
上記化合物が、化学療法薬剤からなる群より選択されることを特徴とする項目5また
は6に記載の使用。
(項目8)
上記化学療法薬剤が、パクリタキセルおよびドセタキセルなどのタキサン、ドキソルビシンなどのアントラサイクリン、ならびにゲムシタビンからなる群より選択されることを特徴とする項目7に記載の使用。
(項目9)
上記化合物が、ADCC(抗体依存性細胞障害作用)によって腫瘍細胞を殺傷する治療抗体と、該治療抗体の混合物とからなる群より選択されることを特徴とする項目5また
は6に記載の使用。
(項目10)
上記治療抗体が、リツキシマブ、セツキシマブ、エドレコロマブおよびトラスツズマブからなる群より選択されることを特徴とする項目9に記載の使用。
(項目11)
同時に、別々に、または連続して使用するための、組換えLAG−3タンパク質またはその誘導体と、項目9または10に記載の治療抗体とを含む、部品のキット。
(項目12)
同時に、別々に、または連続して使用するための、組換えLAG−3タンパク質またはその誘導体と、項目7または8に記載の抗癌性の化学治療特性を有する化合物とを含む
、部品のキット。
<実施例1:低用量IMP321を用いた場合の、転移性乳癌(MBC)患者における単球数増加>
腫瘍細胞のアポトーシスを誘導することで知られている化学療法を受けている5人のMBC患者に対して、それぞれ次のようにIMP321の皮下投与を行なった。すなわち、1回につき0.25mgのIMP321を、隔週で行なわれる化学療法を受けてから1〜2日後に皮下注射にて投与する。以上のような方法で、24週にわたって投与を行なった。各IMP321は14日間あけて投与される。
えた。
3人のMRCC患者に対して、6.25mgのIMP321を隔週で皮下注射にて投与した。投与期間は12週間で、各投与は14日の間隔をおいて行なわれた。
乳癌に対する一次化学療法として、1回28日サイクルの1日目、8日目および15日目にパクリタキセル(80mg/m2を静注で投与)の投与を受けるサイクルを6サイクル受ける患者に対して、各28日サイクルの2日目および16日目に1〜30mgのIMP321の皮下投与を行なった。IMP321は、3日目または17日目に投与してもよい。
進行性膵臓癌(あるいは腫瘍の摘出手術を受けられない患者)の一次化学療法として、1回28日サイクルの1日目、8日目および15日目に標準的なゲムシタビン投与(1gm/m2を30分にわたり静注で投与)を受けるサイクルを6サイクル受ける患者に対して、各28日サイクルの2日目および16日目に6〜30mgのIMP321の皮下投与を行なった。IMP321は、3日目または17日目に投与してもよい。
まず、PBMCを、IMP321とともにまたはIMP321を含めずに(IMP321の濃度を、0μg/m、0.03μg/mlまたは0.1μg/mlとして)、IL−2(100U/ml)とともに40時間培養する。その後、PBMCを、標的分子(すなわち、ヒトCD20+ラジB細胞)の存在下で、濃度が増加傾向にあるリツキシマブ(0μg/ml、0.5μg/mlおよび5μg/ml)とともにインキュベートする。
[1]TRIEBELet. al.,J. Exp. Med.,171:1393-1405,1990
[2]BRIGNONEet. al., J. Immune Based Ther Immunotherapies, 5: 5, 2007
[3]TRIEBELet. al., Trends Immunol., 24: 619-622, 2003
[4]HUARDet.al., Proc. Natl. Acad. Sci. USA, 94: 5744-5749, 1997
[5]PRIGENTet. al., Eur. J. Immunol., 29: 3867-3876, 1999
[6]BRIGNONEet. al., J. Immunol., 179: 4202-4211, 2007。
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