JP2011105749A - 組み合わせ免疫賦活薬 - Google Patents
組み合わせ免疫賦活薬 Download PDFInfo
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- JP2011105749A JP2011105749A JP2011015347A JP2011015347A JP2011105749A JP 2011105749 A JP2011105749 A JP 2011105749A JP 2011015347 A JP2011015347 A JP 2011015347A JP 2011015347 A JP2011015347 A JP 2011015347A JP 2011105749 A JP2011105749 A JP 2011105749A
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Abstract
【解決手段】一般に組み合わせ免疫賦活薬は、TLR作動薬およびTNF/R作動薬を含む。特定の組み合わせ免疫賦活薬はまた、抗原を含んでも良い。
【選択図】なし
Description
(a)例えばアデノウィルス、ヘルペスウィルス(例えばHSV−I、HSV−II、CMV、またはVZV)、ポックスウィルス(例えば痘瘡またはワクシニア、または伝染性軟属腫などのオルトポックスウィルス)、ピコルナウィルス(例えばライノウィルスまたはエンテロウィルス)、オルソミクソウィルス(例えばインフルエンザウィルス)、パラミクソウィルス(例えばパラインフルエンザウィルス、おたふく風邪ウィルス、はしかウィルス、呼吸器合胞体ウイルス(RSV))、コロナウィルス(例えばSARS)、パポバウィルス(例えば生殖器疣、尋常性肬贅、または足底疣贅を引き起こすものなどの乳頭腫ウィルス)、ヘパドナウィルス(例えば肝炎Bウィルス)、フラビウイルス(例えば肝炎Cウィルスまたはデングウィルス)、またはレトロウィルス(例えばHIVなどのレンチウイルス)による感染から罹る疾患などのウィルス疾患、
(b)例えばエシェリキア属、エンテロバクター、サルモネラ、ブドウ球菌、赤痢菌、リステリア、アエロバクター、ヘリコバクター、クレブシエラ、プロテウス、シュードモナス、連鎖球菌、クラミジア、マイコプラズマ、肺炎球菌、ナイセリア、クロストリジウム、バシラス、コリネバクテリウム、マイコバクテリウム、カンピロバクター、ビブリオ、セラチア、プロビデンシア、クロモバクテリウム、ブルセラ、エルシニア、ヘモフィルス、またはボルデテラなどの細菌感染から罹る疾患などの細菌疾患、
(c)クラミジアと、カンジダ症、アスペルギルス症、ヒストプラスマ症、クリプトコックス髄膜炎をはじめとするがこれに限定されるものではない真菌疾患と、またはマラリア、ニューモシステイスカリニ肺炎、レーシュマニア症、クリプトスポリジウム症、トキソプラズマ症、およびトリパノソーマ感染をはじめとするがこれに限定されるものではない寄生虫性疾患などのその他の感染性疾患、
(d)例えば上皮内新生物形成、子宮頚部異形成、光線性角化症、基底細胞癌、扁平細胞癌、腎細胞癌、カポジ肉腫、メラノーマ、腎細胞癌などの新生物疾患、骨髄性白血病、慢性リンパ性白血病、多発性骨髄腫、非ホジキンリンパ腫、皮膚T−細胞リンパ腫、B−細胞リンパ腫、および毛様細胞白血病をはじめとするが、これに限定されるものではない白血病、およびその他の癌(例えば上で同定された癌)、および
(e)アトピー性皮膚炎または湿疹、好酸球増加症、喘息、アレルギー、アレルギー性鼻炎、全身性エリテマトーデス、本態性血小板血症、多発性硬化症、オメン症候群、円板状エリテマトーデス、円形脱毛症などのTH2媒介アトピー性および自己免疫疾患と、ケロイド形成およびその他のタイプの瘢痕の阻害と、慢性創傷をはじめとする創傷治癒の増進などであるが、これに限定されるものではない病状を処置するのに有用であることを示唆する。
2〜5匹のマウスを(A)100μgの卵白アルブミンペプチド、(B)100μgの卵白アルブミンペプチド+100μgの抗−CD40抗体(1C10)、(C)100μgの卵白アルブミンペプチド+200μgのIRM1、または(D)100μgの卵白アルブミンペプチド+100μgの1C10抗−CD40抗体+200μgのIRM1で静脈内免疫した。免疫化の5日後、マウスから脾臓を切除して均質化した。均質化した細胞懸濁液を卵白アルブミン−特異的T細胞(Kedlら、JEM 192(8):1105〜1113(2000))を検出するために主要組織適合性複合体(MHC)四量体試薬で染色し、カリフォルニア州サンディエゴのBDバイオサイエンシズ・ファーミンゲン(BD Biosciences Pharmingen(San Diego、CA))からのCD8染色、およびバイオサイエンシズ・ファーミンゲン(BD Biosciences Pharmingen(San Diego、CA))からのCD44染色で染色した。フローサイトメトリーにかけると、卵白アルブミン−特異的CD8+T細胞は、図1に示すドットプロットの右上4分の1区中に示される。抗−CD40抗体およびIRMの組み合わせによる刺激後の卵白アルブミン−特異的CD8+T細胞集団の増殖は、抗−CD40抗体またはIRMいずれか単独での刺激後の卵白アルブミン−特異的CD8+T細胞集団の増殖を超えた。
マウスに、ミズーリ州セントルイスのシグマ・ケミカル(Sigma Chemical Co.(St.Louis、MO))からの卵白アルブミン5mg、50μgのFGK4.5抗−CD40抗体、および220μgのIRM1を腹腔内注射した。マウスをそれぞれ4、5、6、9、および12日目に屠殺した。屠殺マウスから脾臓を切除して均質化した。実施例1で述べるようにして、均質化した細胞懸濁液を染色し分析した。フローサイトメトリーにかけると、卵白アルブミン−特異的CD8+T細胞(上)および卵白アルブミン−特異的CD8+/CD44+T細胞(下)が同定され、各ドットブロットの右上4分の1区中に示される。右上4分の1区中の数値は、その4分の1区中の細胞の百分率を示す。これらのデータは、実施例1で観察されたCD8+T細胞増殖における相乗効果が、(a)異なるCD40作動薬、および(b)抗原としてのフルサイズの卵白アルブミンタンパク質によっても観察されたことを示す。
マウスを100μgのFGK4.5抗−CD40抗体+200μgのIRM1と、(A)ペプチドなし、(B)100μgの卵白アルブミンペプチド、または(C)100μgのTRP2−ΔVペプチドのいずれかとで静脈内免疫した。免疫化の5日後、マウスから脾臓を切除して均質化した。TRP2−ΔV−特異的T細胞を検出するためにMHC四量体試薬を調製したこと以外は、実施例1のようにして均質化した細胞懸濁液を染色した。フローサイトメトリーにかけると、TRP2−ΔV−特異的CD8+T細胞は、図3に示すドットブロットの右上4分の1区中に示される。右上4分の1区中の数値は、その4分の1区中の細胞の百分率を示す。データは、抗−CD40抗体およびIRMとさらに別の抗原との組み合わせによる刺激後の抗原−特異的CD8+T細胞の相乗的増殖を示す。
0日目に、マウスを100μgの卵白アルブミンペプチド+200μgのIRM1+100μgの1C10抗−CD40抗体で静脈内免疫した。28日目に、マウスを(A)未チャレンジ、(B)100μgの卵白アルブミンペプチドで静脈内チャレンジ、または(C)100μgの卵白アルブミンペプチド+200μgのIRM1で静脈内チャレンジした。33日目にマウスを屠殺して、脾臓を切除して脾臓細胞を均質化した。実施例1で述べるようにして、均質化した細胞を染色し分析した。データを図4に示す。抗原、CD40作動薬、およびTLR作動薬(実施例1に示す)による免疫化の結果として起きたCD8+T細胞の相乗的増殖は、(C)に示すIRMおよび抗原での処置により再活性化できる長命のCD8+メモリT細胞のプールを産生した。
表2に示すように、マウスを静脈内免疫した。5日目にマウスを屠殺して脾臓を採取し、実施例1のようにして細胞を均質化して染色し分析した。データを図5に示す。右上4分の1区中の数値は、その4分の1区における細胞の百分率を示す。
0日目に、マウスをPBS中の1×105メラノーマB16卵巣腫瘍細胞で皮内チャレンジした(ケドル(Kedl)らPNAS 98(19):10811〜10816)。7日目に、マウスを(A)100μgの卵白アルブミンペプチド、(B)100μgの卵白アルブミンペプチド+200μgのIRM1、または(C)100μgの卵白アルブミンペプチド+200μgのIRM1+100μgの1C10抗−CD40抗体のいずれかによって免疫した。21日目にマウスを屠殺して、ノギスによって腫瘍を二次元で測定した。データを図6に示す。抗原、IRM、およびCD40作動薬による免疫化は、IRM単独での免疫化よりも緩慢な腫瘍生育をもたらした。
実施例6のようにして、マウスを0日目に腫瘍でチャレンジした。5匹のマウスをそれぞれ7日目に、(A)1×106細胞同等物(CE)の腫瘍溶解産物、(B)1×106CE腫瘍溶解産物+200μgのIRM1、(C)1×106CE腫瘍溶解産物+100μgのFGK4.5抗−CD40抗体、または(D)1×106CE腫瘍溶解産物+200μgのIRM1+100μgのFGK4.5抗−CD40抗体で免疫した。14および20日目に、マウスにおいてノギスで腫瘍サイズを測定した。データを図7に示す。IRMおよび抗−CD40作動薬の組み合わせによる免疫化は、IRM単独またはCD40作動薬単独による免疫化よりも緩慢な腫瘍生育をもたらす。
0日目に、マウスに500μgの卵白アルブミン、50μgのCD40作動薬(FGK4.5)と、500μgのIRM3、200μgのIRM4、800μgのIRM5、800μgのIRM2、またはIRMなし(対照)のいずれかとを腹腔内注射した。6日目にマウスを屠殺して、脾臓細胞を採取して実施例2で述べるようにして分析した。図8は、マウス(各群でn=3)の各群で観察されたCD8+T細胞の平均百分率を示す。CD40作動薬を異なるIRM化合物との組み合わせで使用して、CD8+T細胞の相乗的増殖が実証される。
0日目に、マウスを1mgの卵白アルブミン、200μgのIRM1と、200μgのCD40リガンド(FGK4.5)、200μgの4−1BBリガンド(カリフォルニア州サンディエゴのイーバイオサイエンスからの抗−マウス4−1BB抗体、クローン17B3(eBioscience、(San Diego、CA))、またはTNF/R作動薬なし(対照)のいずれかとによって免疫した。6日目に、マウスを屠殺して脾臓細胞を採取し、実施例2で述べるようにして分析した。結果を図9に示す。IRM1を異なるTNF/R作動薬との組み合わせで使用して、CD8+T細胞の相乗的増殖が実証される。
0日目に、一組の野生型マウス(ニューヨーク州ジャーマンタウンのタコニック(Taconic(Germantown、NY))からのB6/129 F1)および一組のIFNαβ受容体ノックアウトマウス(コロラド州デンバーのナショナル・ジュウイッシュ・メディカル・アンド・リサーチセンター(National Jewish Medical and Research Center(Denver、CO))に、100μgのSIINFEKLペプチド、50μgのFGK45(CD40作動薬)と、(a)なし(CD40のみ)、(b)100μgのIRM1(+TLR7)、(c)50μgのポリIC(+TLR3)、100μgのCpG(+TLR9)、30μgのLPS(+TLR4)、または25μgのMALP−2(+TLR2)のいずれかとを腹腔内に注射した。6日目にマウスを屠殺し脾臓細胞を採取して、実施例2で述べるようにして分析した。
一組の野生型マウス(ニューヨーク州ジャーマンタウンのタコニック(Taconic(Germantown、NY))からのB6/129F1)および一組のIFNαβ受容体ノックアウトマウス(コロラド州デンバーのナショナル・ジュウイッシュ・メディカル・アンド・リサーチセンター(National Jewish Medical and Research Center(Denver、CO))に、0日目に50μgのFGK45(CD40作動薬)を腹腔内注射した。4時間後、マウスに100μgのSIINFEKLを単独で、または100μgのIRM1(TLR7作動薬)、25μgのMALP−2、カリフォルニア州サンディエゴのアレクシス・バイオケミカルズ(Alexis Biochemicals、Corp.(San Diego、CA))からの50μgのPam3cys、100μgのPam3cys、または250μgのPam3cysのいずれかを静脈注射した。6日目にマウスを屠殺し脾臓細胞を採取して、実施例2で述べるようにして分析した。結果を図11に示す。TLR2/6作動薬であるMALP−2を含む組み合わせ免疫賦活薬によって観察されたインターフェロン−非依存性相乗的免疫応答は、TLR2作動薬であるPam3cysを含む組み合わせ免疫賦活薬を使用しても観察された。
Claims (57)
- TLR作動薬およびTNF/R作動薬を、それぞれ、被験者の抗原に対する免疫応答を増大させるのに他方との組み合わせにおいて有効な量で含む、組み合わせ免疫賦活薬。
- 上記TLR作動薬が、TLR1、TLR2、TLR3、TLR4、TLR5、TLR6、TLR7、TLR8、TLR9、TLR10、または前述の任意の作動薬の任意の組み合わせ、の少なくとも1つの作動薬である、請求項1に記載の組み合わせ免疫賦活薬。
- 上記TLR作動薬が、IRM化合物またはTLR2の作動薬を含む、請求項2に記載の組み合わせ免疫賦活薬。
- 上記TLR作動薬が、IRM化合物、MALP−2、LPS、ポリIC、CpG、または前述の任意のものの任意の組み合わせを含む、請求項1に記載の組み合わせ免疫賦活薬。
- 上記IRM化合物が、イミダゾキノリンアミン、テトラヒドロイミダゾキノリンアミン、イミダゾピリジンアミン、1,2−架橋イミダゾキノリンアミン、6,7−縮合シクロアルキルイミダゾピリジンアミン、イミダゾナフチリジンアミン、テトラヒドロイミダゾナフチリジンアミン、オキサゾロキノリンアミン、チアゾロキノリンアミン、オキサゾロピリジンアミン、チアゾロピリジンアミン、オキサゾロナフチリジンアミン、またはチアゾロナフチリジンアミンを含む、請求項3に記載の組み合わせ免疫賦活薬。
- 上記TNF/R作動薬が、TNFスーパーファミリーメンバーの作動薬を含む、請求項1に記載の組み合わせ免疫賦活薬。
- 上記TNF/R作動薬が、CD40リガンド、OX40リガンド、4−1BBリガンド、CD27、CD30リガンド(CD153)、TNF−α、TNF−β、RANKリガンド、LT−α、LT−β、GITRリガンド、またはLIGHTの作動薬を含む、請求項6に記載の組み合わせ免疫賦活薬。
- 上記TNF/R作動薬が、TNFRスーパーファミリーメンバーの作動薬を含む、請求項1に記載の組み合わせ免疫賦活薬。
- 上記TNF/R作動薬が、CD40、OX40、4−1BB、CD70(CD27リガンド)、CD30、TNFR2、RANK、LT−βR、HVEM、GITR、TROY、またはRELTの作動薬を含む、請求項8に記載の組み合わせ免疫賦活薬。
- 上記TNF/R作動薬が、作動薬抗体を含む、請求項1に記載の組み合わせ免疫賦活薬。
- 被験者に、TLR作動薬およびTNF/R作動薬を、それぞれ、TH1免疫応答を誘導するのに他方との組み合わせにおいて有効な量で同時投与するステップを含む、被験者においてTH1免疫応答を誘導する方法。
- 上記TLR作動薬がTLR2の作動薬を含む、請求項11に記載の方法。
- 上記TLR作動薬がTLR9の作動薬を含む、請求項11に記載の方法。
- 上記TLR作動薬がTLR8の作動薬を含む、請求項11に記載の方法。
- 上記TLR作動薬がTLR7の作動薬を含む、請求項11に記載の方法。
- 被験者において免疫応答を誘導するのに有効な量で、抗原を同時投与するステップをさらに含む、請求項11に記載の方法。
- 被験者に、TLR作動薬およびTNF/R作動薬を、それぞれ、抗原特異的CD8+T細胞を活性化するのに他方との組み合わせにおいて有効な量で同時投与するステップを含む、被験者において抗原特異的CD8+T細胞を活性化する方法。
- 抗原を被験者において免疫応答を誘導するのに有効な量で同時投与するステップをさらに含む、請求項17に記載の方法。
- CD8+T細胞の活性化が、CD8+エフェクターT細胞の増殖を含む、請求項17に記載の方法。
- CD8+T細胞の活性化が、CD8+メモリT細胞の産生を含む、請求項17に記載の方法。
- 上記TLR作動薬がTLR2の作動薬を含む、請求項17に記載の方法。
- 上記TLR作動薬がTLR9の作動薬を含む、請求項17に記載の方法。
- 上記TLR作動薬がTLR8の作動薬を含む、請求項17に記載の方法。
- 上記TLR作動薬がTLR7の作動薬を含む、請求項17に記載の方法。
- 上記TNF/R作動薬が、TNFスーパーファミリーメンバーの作動薬を含む、請求項17に記載の方法。
- 上記TNF/R作動薬が、TNFRスーパーファミリーメンバーの作動薬を含む、請求項17に記載の方法。
- 上記TNF/R作動薬が作動薬抗体を含む、請求項17に記載の方法。
- 抗原を抗原−特異的CD8+メモリT細胞を活性化させるのに有効な量で被験者に投与することによって、抗原−特異的CD8+エフェクターT細胞を産生するステップを含む、抗原にあらかじめ曝露された上記被験者において抗原−特異的メモリCD8+T細胞を活性化する方法。
- TLR作動薬を抗原−特異的CD8+メモリT細胞を活性化させるのに有効な量で同時投与することで、抗原−特異的CD8+エフェクターT細胞を産生するステップをさらに含む、請求項28に記載の方法。
- 上記TLR作動薬がTLR2の作動薬を含む、請求項28に記載の方法。
- 上記TLR作動薬がTLR9の作動薬を含む、請求項28に記載の方法。
- 上記TLR作動薬がTLR8の作動薬を含む、請求項28に記載の方法。
- 上記TLR作動薬がTLR7の作動薬を含む、請求項28に記載の方法。
- 他方との組み合わせにおいて、細胞媒介免疫応答を刺激するのに有効な量でそれぞれ投与されるTLR作動薬およびTNF/R作動薬を被験者に同時投与するステップを含む、上記被験者において病状を処置する方法。
- 上記TLR作動薬がTLR2の作動薬を含む、請求項34に記載の方法。
- 上記TLR作動薬がTLR9の作動薬を含む、請求項34に記載の方法。
- 上記TLR作動薬がTLR8の作動薬を含む、請求項34に記載の方法。
- 上記TLR作動薬がTLR7の作動薬を含む、請求項34に記載の方法。
- 上記TNF/R作動薬が、TNFスーパーファミリーメンバーの作動薬を含む、請求項34に記載の方法。
- 上記TNF/R作動薬が、TNFRスーパーファミリーメンバーの作動薬を含む、請求項34に記載の方法。
- 上記TNF/R作動薬が作動薬抗体を含む、請求項34に記載の方法。
- 病状に関連する抗原を細胞媒介免疫応答を誘導するのに有効な量で同時投与するステップをさらに含む、請求項34に記載の方法。
- 上記病状が新生物疾患を含む、請求項34に記載の方法。
- 上記TLR作動薬および上記TNF/R作動薬を同時投与するステップが、予防的な処置を提供する、請求項43に記載の方法。
- 上記TLR作動薬および上記TNF/R作動薬を同時投与するステップが、治療的な処置を提供する、請求項43に記載の方法。
- 上記病状が感染性疾患を含む、請求項34に記載の方法。
- 上記TLR作動薬および上記TNF/R作動薬を同時投与するステップが、予防的な処置を提供する、請求項46に記載の方法。
- 上記TLR作動薬および上記TNF/R作動薬を同時投与するステップが、治療的な処置を提供する、請求項46に記載の方法。
- ワクチンであって、TLR作動薬、TNF/R作動薬、および抗原を、それぞれ、前記ワクチンで免疫された被験者において抗原に対する免疫応答を誘導するのに他のものとの組み合わせにおいて有効な量で含むワクチン。
- 上記TLR作動薬がTLR2の作動薬を含む、請求項49に記載のワクチン。
- 上記TLR作動薬がTLR9の作動薬を含む、請求項49に記載のワクチン。
- 上記TLR作動薬がTLR8の作動薬を含む、請求項49に記載のワクチン。
- 上記TLR作動薬がTLR7の作動薬を含む、請求項49に記載のワクチン。
- 上記TNF/R作動薬が、TNFスーパーファミリーメンバーの作動薬を含む、請求項49に記載のワクチン。
- 上記TNF/R作動薬が、TNFRスーパーファミリーメンバーの作動薬を含む、請求項49に記載のワクチン。
- 上記TNF/R作動薬が作動薬抗体を含む、請求項49に記載のワクチン。
- 上記抗原が、腫瘍抗原、ウィルス抗原、細菌抗原、または寄生虫抗原を含む、請求項49に記載のワクチン。
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EP1578419A2 (en) | 2005-09-28 |
US20110280903A1 (en) | 2011-11-17 |
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US10105426B2 (en) | 2018-10-23 |
JP2011016811A (ja) | 2011-01-27 |
US20160129095A1 (en) | 2016-05-12 |
JP2014101371A (ja) | 2014-06-05 |
AU2003300184B2 (en) | 2009-08-06 |
WO2004060319A3 (en) | 2004-11-04 |
US20190083592A1 (en) | 2019-03-21 |
US20090123460A1 (en) | 2009-05-14 |
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