JP2021503503A - リソソーム誘発性免疫原性細胞死のシステムおよび方法 - Google Patents
リソソーム誘発性免疫原性細胞死のシステムおよび方法 Download PDFInfo
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Abstract
Description
酸トリメトレキセート、トリプトレリン、トロピセトロン、塩酸ツブロゾール、ツロステリド、チロシンキナーゼ阻害剤、チロホスチン、UBC阻害剤、ウベニメクス、ウラシルマスタード、ウレデパ、泌尿生殖洞由来成長阻害因子、ウロキナーゼ受容体拮抗薬、バプレオチド、バリオリンB、ベラレゾール、ベラミン、ベルジン、ベルテポルフィン、硫酸ビンブラスチン、ビンクリスチン硫酸、ビンデシン、硫酸ビンデシン、硫酸ビネピジン、硫酸ビグリシネート、硫酸ビンロイロシン、ビノレルビン、酒石酸ビノレルビン、硫酸ビンロシジン、ビンキサルチン、硫酸ビンゾリジン、ビタキシン、ボロゾール、ザノテロン、ゼニプラチン、ジラスコルブ、ジノスタチン、ジノスタチンスティマラマー、および塩酸ゾルビシン、ならびにそれらの塩、同族体、類似体、多形、誘導体、エナンチオマー、および/または機能的に等価な組成物のいずれか1つまたは複数が含まれるが、これらに限定されない。
免疫原性細胞死(ICD)への進行における1つのステップは、ネオ抗原の生成である。リソソームには、プロテアーゼ、グリコシダーゼ、ホスファターゼなどのさまざまな分解酵素が含まれている。これらの酵素は細胞質に放出された場合、細胞内の内容物を分解し、その過程でネオ抗原を生成する。次に、これらネオ抗原は樹状細胞およびマクロファージによって取り込まれる。これらの細胞はリンパ節に移動し、そこでネオ抗原がT細胞およびB細胞を活性化し、T細胞およびB細胞は適応免疫系の活性化によって細胞死を引き起こす。
この予言的な実施例では、腫瘍を有するヒト対象に対して、腫瘍を認識する抗体および酵素を含む複合体を含む組成物が提供される。場合によっては、抗体は、例えば腫瘍認識抗体またはモノクローナル抗体など、商業的に入手可能なものである。
この予言的な実施例では、ヒト対象は実施例2のように治療されるが、さらに、対象は、複合体の作用を妨害する抗酸化物質を離脱および/または抑制するための治療処置計画にさらされる。一例では、対象は、CH2=CHCOOHなどの不飽和脂肪酸を提供される。脂肪酸は、抗酸化剤の作用を阻害するように作用し、複合体に対するより大きな反応をもたらす。
この予言的な実施例では、ヒト対象は実施例2のように治療されるが、さらに、対象は、アトルバスタチンなどのスタチンにさらされ、スタチンは抗酸化物質の作用を阻害するように作用し、複合体に対するより大きな反応をもたらす。
この予言的な実施例では、epCAM認識抗体と酵素キサンチンオキシダーゼの間のコンジュゲートが、アミノからアミノへの架橋剤を使用して調製される。抗epCAMによって認識されるMC26悪性細胞を96ウェルプレートで培養し、10マイクログラム/ml〜100マイクログラム/mlの適切な濃度のコンジュゲートで処理する。ヒポキサンチンは、1mMの濃度で細胞培養培地に提供された。ICDの認められた特徴がある。これらには、カルレティキュリン曝露、細胞外ATP形成、およびHMGB1形成が含まれる。これらは1時間、2時間、4時間、6時間、12時間および24時間でアッセイされる。結果のグラフを図3に示す。適切にカルレティキュリンが形成され、その後にATP形成が続き、その後にHMGB1形成が続く。これは、細胞がICDのマーカーを産生し、T細胞を活性化して腫瘍に誘導するネオ抗原を死ぬ過程で形成すると予想されることを確立する。
ICDを誘発するプロセスで腫瘍が殺されると、腫瘍を有する動物はその腫瘍に対して「免疫」される。したがって、ICDは将来的にその腫瘍から動物を保護する。
活性酸素種(ROS)は、リソソーム膜透過性(LMP)を引き起こし、リソソーム酵素の放出およびアポトーシスによる細胞死を引き起こすプロセスを開始できることが示されている。ROS、特にスーパーオキシドの供給源は、腫瘍微小環境を含む細胞外および/または細胞内であり得る。両者の結果は、リソソームとその結果生じるアポトーシスに関して同じであるが、メカニズムは多少異なる。この実施例では、これらの2つのメカニズムとその一般的な結果、ICDに必要なネオ抗原を産生する一次細胞死(Primary Cell Death; PCD)について説明する。
Claims (26)
- 以下を含む物品:
腫瘍を認識する抗体と、リソソームの膜透過性を増加させることができる酵素とを含む複合体。 - 以下を含む物品:
腫瘍を認識する抗体と、活性酸素種を生成することができる酵素とを含む複合体。 - 酵素がオキシダーゼである、請求項1または2のいずれか一項に記載の物品。
- オキシダーゼがグルコースオキシダーゼである、請求項3に記載の物品。
- オキシダーゼがキサンチンオキシダーゼである、請求項3に記載の物品。
- 酵素がペルオキシダーゼである、請求項1または2のいずれか一項に記載の物品。
- 抗体および酵素が直接共有結合している、請求項1〜6のいずれか一項に記載の物品。
- 抗体および酵素が連結剤を介して結合される、請求項1〜7のいずれか一項に記載の物品。
- 以下を含む方法:
対象に、腫瘍を認識する抗体と、リソソームの膜透過性を増加させることができる酵素との複合体を含む組成物を投与する。 - 以下を含む方法:
対象に、腫瘍を認識する抗体と、活性酸素種を生成することができる酵素との複合体を含む組成物を投与する。 - 以下を含む方法:
対象に、腫瘍を認識する抗体と、リソソーム漏出を引き起こすことができる酵素との複合体を含む組成物を投与する。 - 酵素がオキシダーゼである、請求項9〜11のいずれか一項に記載の方法。
- オキシダーゼがグルコースオキシダーゼである、請求項12に記載の方法。
- オキシダーゼがキサンチンオキシダーゼである、請求項12に記載の方法。
- 酵素がペルオキシダーゼである、請求項9〜11のいずれか一項に記載の方法。
- 抗体および酵素が直接共有結合している、請求項9〜15のいずれか一項に記載の方法。
- 抗体および酵素が連結剤を介して結合される、請求項9〜16のいずれか一項に記載の方法。
- 脂肪酸を対象に投与することをさらに含む、請求項9〜17のいずれか一項に記載の方法。
- 脂肪酸が不飽和C3-C6脂肪酸を含む、請求項18に記載の方法。
- 組成物が脂肪酸を含む、請求項18または19のいずれか一項に記載の方法。
- 複合体の投与と脂肪酸の投与が同時に起こる、請求項18〜20のいずれか一項に記載の方法。
- 複合体の投与と脂肪酸の投与が連続して行われる、請求項18〜21のいずれか一項記載の方法。
- 対象にスタチンを投与することをさらに含む、請求項8〜22のいずれか一項に記載の方法。
- 組成物がスタチンを含む、請求項23に記載の方法。
- 複合体の投与とスタチンの投与が同時に行われる、請求項23または24のいずれか一項に記載の方法。
- 複合体の投与とスタチンの投与が連続して行われる、請求項23〜25のいずれか一項に記載の方法。
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PCT/US2018/061304 WO2019099687A1 (en) | 2017-11-16 | 2018-11-15 | Systems and methods for lysosome induced immunogenic cell death |
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