JP2014169279A - 経皮投与用wt1ペプチド癌ワクチンテープ剤 - Google Patents
経皮投与用wt1ペプチド癌ワクチンテープ剤 Download PDFInfo
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- JP2014169279A JP2014169279A JP2014014805A JP2014014805A JP2014169279A JP 2014169279 A JP2014169279 A JP 2014169279A JP 2014014805 A JP2014014805 A JP 2014014805A JP 2014014805 A JP2014014805 A JP 2014014805A JP 2014169279 A JP2014169279 A JP 2014169279A
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Abstract
【解決手段】本発明は、(i)WT1ペプチドおよび/または改変WT1ペプチド;ならびに(ii)細胞性免疫誘導促進剤を含む、細胞性免疫誘導のための経皮投与用癌ワクチンテープ剤を提供する。
【選択図】図1
Description
(1)(i)WT1ペプチドおよび/または改変WT1ペプチド;ならびに
(ii)TLRリガンド、環状ジヌクレオチド、ヘルパーペプチド、免疫調節低分子薬物、シクロオキシゲナーゼ阻害剤、プロスタグランジン受容体拮抗薬、プロスタグランジン受容体作動薬、TSLP産生抑制剤、アデニル酸シクラーゼ阻害剤、オメガ3脂肪酸、PPAR作動薬、ドーパミン受容体拮抗薬、ドーパミン受容体作動薬、ヒスタミン受容体作動薬、ヒスタミン受容体拮抗薬、セロトニン受容体作動薬、セロトニン受容体拮抗薬、バソプレシン受容体拮抗薬、バソプレシン受容体作動薬、ムスカリン受容体拮抗薬、ムスカリン受容体作動薬、アドレナリン受容体拮抗薬、アドレナリン受容体作動薬、アンジオテンシン受容体作動薬、GABA受容体作動薬、トロンビン受容体拮抗薬、トロンビン受容体作動薬、オピオイド受容体作動薬、ADP受容体作動薬、ロイコトリエン受容体拮抗薬、ロイコトリエン受容体作動薬、メラトニン受容体作動薬、ソマトスタチン受容体作動薬、カンナビノイド受容体作動薬、スフィンゴシン1リン酸受容体作動薬、代謝型グルタミン酸受容体作動薬、ホスホリパーゼA2阻害剤、TGF−β産生抑制剤、Th2サイトカイン阻害剤、およびそれらの2種以上の組合せから選択される第一の細胞性免疫誘導促進剤
を含む、細胞性免疫誘導のための経皮投与用癌ワクチンテープ剤;
(2)さらに第二の細胞性免疫誘導促進剤としての薬理学的に許容される酸またはその薬理学的に許容される塩を含む、(1)の経皮投与用癌ワクチンテープ剤;
(3)第一の細胞性免疫誘導促進剤がTLRリガンドである、(1)または(2)の経皮投与用癌ワクチンテープ剤;
(4)第一の細胞性免疫誘導促進剤が環状ジヌクレオチドである、(1)または(2)の経皮投与用癌ワクチンテープ剤;
(5)第一の細胞性免疫誘導促進剤が免疫調節低分子薬物である、(1)または(2)の経皮投与用癌ワクチンテープ剤;
(6)第一の細胞性免疫誘導促進剤がシクロオキシゲナーゼ阻害剤である、(1)または(2)の経皮投与用癌ワクチンテープ剤;
(7)第一の細胞性免疫誘導促進剤がプロスタグランジン受容体拮抗薬であり、更にプロスタグランジン受容体拮抗薬が、EP2受容体拮抗薬、EP4受容体拮抗薬、DP受容体拮抗薬、IP受容体拮抗薬である、(1)または(2)の経皮投与用癌ワクチンテープ剤;
(8)第一の細胞性免疫誘導促進剤がプロスタグランジン受容体作動薬であり、更にプロスタグランジン受容体作動薬が、EP3受容体作動薬である、(1)または(2)の経皮投与用癌ワクチンテープ剤;
(9)第一の細胞性免疫誘導促進剤がTSLP産生抑制剤である、(1)または(2)の経皮投与用癌ワクチンテープ剤;
(10)第一の細胞性免疫誘導促進剤がアデニル酸シクラーゼ阻害剤である、(1)または(2)の経皮投与用癌ワクチンテープ剤;
(11)第一の細胞性免疫誘導促進剤がオメガ3脂肪酸である、(1)または(2)の経皮投与用癌ワクチンテープ剤;
(12)第一の細胞性免疫誘導促進剤がPPAR作動薬であり、更にPPAR作動薬がPPAR−α作動薬、PPAR−δ作動薬、PPAR−γ作動薬である(1)または(2)の経皮投与用癌ワクチンテープ剤;
(13)第一の細胞性免疫誘導促進剤がドーパミン受容体拮抗薬であり、更にドーパミン受容体拮抗薬が、D1受容体拮抗薬、D5受容体拮抗薬である、(1)または(2)の経皮投与用癌ワクチンテープ剤;
(14)第一の細胞性免疫誘導促進剤がドーパミン受容体作動薬であり、更にドーパミン受容体作動薬が、D2受容体作動薬、D3受容体作動薬、D4受容体作動薬である、(1)または(2)の経皮投与用癌ワクチンテープ剤;
(15)第一の細胞性免疫誘導促進剤がヒスタミン受容体拮抗薬であり、更にヒスタミン受容体拮抗薬が、H1受容体拮抗薬、H2受容体拮抗薬である、(1)または(2)の経皮投与用癌ワクチンテープ剤;
(16)第一の細胞性免疫誘導促進剤がヒスタミン受容体作動薬であり、更にヒスタミン受容体作動薬が、H1受容体作動薬、H3受容体作動薬、H4受容体作動薬である、(1)または(2)の経皮投与用癌ワクチンテープ剤;
(17)第一の細胞性免疫誘導促進剤がセロトニン受容体拮抗薬であり、更にセロトニン受容体拮抗薬が、5−HT2受容体拮抗薬、5−HT4受容体拮抗薬、5−HT6受容体拮抗薬、5−HT7受容体拮抗薬である、(1)または(2)の経皮投与用癌ワクチンテープ剤;
(18)第一の細胞性免疫誘導促進剤がセロトニン受容体作動薬であり、更にセロトニン受容体作動薬が、5−HT1受容体作動薬、5−HT2受容体作動薬である、(1)または(2)の経皮投与用癌ワクチンテープ剤;
(19)第一の細胞性免疫誘導促進剤がバソプレシン受容体拮抗薬であり、更にバソプレシン受容体拮抗薬が、V2受容体拮抗薬である、(1)または(2)の経皮投与用癌ワクチンテープ剤;
(20)第一の細胞性免疫誘導促進剤がバソプレシン受容体作動薬であり、更にバソプレシン受容体作動薬が、V1受容体作動薬である、(1)または(2)の経皮投与用癌ワクチンテープ剤;
(21)第一の細胞性免疫誘導促進剤がムスカリン受容体拮抗薬であり、更にムスカリン受容体拮抗薬が、M1受容体拮抗薬、M3受容体拮抗薬、M5受容体拮抗薬である、(1)または(2)の経皮投与用癌ワクチンテープ剤;
(22)第一の細胞性免疫誘導促進剤がムスカリン受容体作動薬であり、更にムスカリン受容体作動薬が、M1受容体作動薬、M2受容体作動薬、M3受容体作動薬、M4受容体作動薬、M5受容体作動薬である、(1)または(2)の経皮投与用癌ワクチンテープ剤;
(23)第一の細胞性免疫誘導促進剤がアドレナリン受容体拮抗薬であり、更にアドレナリン受容体拮抗薬が、α1受容体拮抗薬、β1受容体拮抗薬、β2受容体拮抗薬、β3受容体拮抗薬である、(1)または(2)の経皮投与用癌ワクチンテープ剤;
(24)第一の細胞性免疫誘導促進剤がアドレナリン受容体作動薬であり、更にアドレナリン受容体作動薬が、α1受容体作動薬、α2受容体作動薬である、(1)または(2)の経皮投与用癌ワクチンテープ剤;
(25)第一の細胞性免疫誘導促進剤がアンジオテンシン受容体作動薬であり、更にアンジオテンシン受容体作動薬が、AT2受容体作動薬である、(1)または(2)の経皮投与用癌ワクチンテープ剤;
(26)第一の細胞性免疫誘導促進剤がGABA受容体作動薬であり、更にGABA受容体作動薬が、GABAB受容体作動薬である、(1)または(2)の経皮投与用癌ワクチンテープ剤;
(27)第一の細胞性免疫誘導促進剤がトロンビン受容体拮抗薬であり、更にトロンビン受容体拮抗薬が、PAR−1受容体拮抗薬である、(1)または(2)の経皮投与用癌ワクチンテープ剤;
(28)第一の細胞性免疫誘導促進剤がトロンビン受容体作動薬であり、更にトロンビン受容体作動薬が、PAR−1受容体作動薬である、(1)または(2)の経皮投与用癌ワクチンテープ剤;
(29)第一の細胞性免疫誘導促進剤がオピオイド受容体作動薬である、(1)または(2)の経皮投与用癌ワクチンテープ剤;
(30)第一の細胞性免疫誘導促進剤がロイコトリエン受容体拮抗薬であり、更にロイコトリエン受容体拮抗薬が、CysLT1受容体拮抗薬、CysLT2受容体拮抗薬である、(1)または(2)の経皮投与用癌ワクチンテープ剤;
(31)第一の細胞性免疫誘導促進剤がロイコトリエン受容体作動薬であり、更にロイコトリエン受容体作動薬が、BLT受容体作動薬である、(1)または(2)の経皮投与用癌ワクチンテープ剤。
(32)第一の細胞性免疫誘導促進剤がメラトニン受容体作動薬である、(1)または(2)の経皮投与用癌ワクチンテープ剤。
(33)第一の細胞性免疫誘導促進剤がソマトスタチン受容体作動薬である、(1)または(2)の経皮投与用癌ワクチンテープ剤。
(34)第一の細胞性免疫誘導促進剤がカンナビノイド受容体作動薬である、(1)または(2)の経皮投与用癌ワクチンテープ剤。
(35)第一の細胞性免疫誘導促進剤がスフィンゴシン1リン酸受容体作動薬である、(1)または(2)の経皮投与用癌ワクチンテープ剤。
(36)第一の細胞性免疫誘導促進剤が代謝型グルタミン酸受容体作動薬であり、更に代謝型グルタミン酸受容体作動薬が、mGluR2受容体作動薬、mGluR3受容体作動薬、mGluR4受容体作動薬、mGluR6受容体作動薬、mGluR7受容体作動薬、mGluR8受容体作動薬である、(1)または(2)の経皮投与用癌ワクチンテープ剤。
(37)第一の細胞性免疫誘導促進剤がADP受容体作動薬である、(1)または(2)の経皮投与用癌ワクチンテープ剤;
(38)第一の細胞性免疫誘導促進剤がホスホリパーゼA2阻害剤である、(1)または(2)の経皮投与用癌ワクチンテープ剤;
(39)第一の細胞性免疫誘導促進剤がTGF−β産生抑制剤である、(1)または(2)の経皮投与用癌ワクチンテープ剤;
(40)第一の細胞性免疫誘導促進剤がTh2サイトカイン阻害剤である、(1)または(2)の経皮投与用癌ワクチンテープ剤;
(41)第一の細胞性免疫誘導促進剤がヘルパーペプチドである、(3)〜(40)のいずれか1項の経皮投与用癌ワクチンテープ剤;
(42)第一の細胞性免疫誘導促進剤が、TLRリガンド、環状ジヌクレオチド、免疫調節低分子薬物、シクロオキシゲナーゼ阻害剤、プロスタグランジン受容体拮抗薬、プロスタグランジン受容体作動薬、TSLP産生抑制剤、アデニル酸シクラーゼ阻害剤、オメガ3脂肪酸、PPAR作動薬、ドーパミン受容体拮抗薬、ドーパミン受容体作動薬、ヒスタミン受容体作動薬、ヒスタミン受容体拮抗薬、セロトニン受容体作動薬、セロトニン受容体拮抗薬、バソプレシン受容体拮抗薬、バソプレシン受容体作動薬、ムスカリン受容体拮抗薬、ムスカリン受容体作動薬、アドレナリン受容体拮抗薬、アドレナリン受容体作動薬、アンジオテンシン受容体作動薬、GABA受容体作動薬、トロンビン受容体拮抗薬、トロンビン受容体作動薬、オピオイド受容体作動薬、ADP受容体作動薬、ロイコトリエン受容体拮抗薬、ロイコトリエン受容体作動薬、メラトニン受容体作動薬、ソマトスタチン受容体作動薬、カンナビノイド受容体作動薬、スフィンゴシン1リン酸受容体作動薬、代謝型グルタミン酸受容体作動薬、ホスホリパーゼA2阻害剤、TGF−β産生抑制剤、Th2サイトカイン阻害剤からなる群より選択される1種以上とヘルパーペプチドとの組合せである、(1)または(2)の経皮投与用癌ワクチンテープ剤;
(43)テープ剤の粘着剤がアクリル系粘着剤である、(1)〜(42)のいずれか1項の経皮投与用癌ワクチンテープ剤;
(44)テープ剤の粘着剤がゴム系粘着剤である、(1)〜(42)のいずれか1項の経皮投与用癌ワクチンテープ剤;
(45)テープ剤の粘着剤がシリコーン系粘着剤である、(1)〜(42)のいずれか1項の経皮投与用癌ワクチンテープ剤;
(46)テープ剤の粘着剤層がさらに皮膚透過性増強剤を含む、(43)〜(45)のいずれか1項の経皮投与用癌ワクチンテープ剤;
(47)低刺激条件下で投与されるものである、(1)〜(46)のいずれか1項の経皮投与用癌ワクチンテープ剤;
(48)低刺激条件が、皮膚刺激評価用モデル動物における投与前の経皮水分蒸散量(TEWL)が50g/h・m2以下の条件である、(47)の経皮投与用癌ワクチンテープ剤;および
(49)低刺激条件が、皮膚刺激評価用モデル動物における投与終了時の皮膚内TSLPレベルが10000pg/mgタンパク質以下の条件である、(47)または(48)の経皮投与用癌ワクチンテープ剤。
(50)(i)WT1ペプチドおよび/または改変WT1ペプチド、ならびに(ii)TLRリガンド、環状ジヌクレオチド、ヘルパーペプチド、免疫調節低分子薬物、シクロオキシゲナーゼ阻害剤、プロスタグランジン受容体拮抗薬、プロスタグランジン受容体作動薬、TSLP産生抑制剤、アデニル酸シクラーゼ阻害剤、オメガ3脂肪酸、PPAR作動薬、ドーパミン受容体拮抗薬、ドーパミン受容体作動薬、ヒスタミン受容体作動薬、ヒスタミン受容体拮抗薬、セロトニン受容体作動薬、セロトニン受容体拮抗薬、バソプレシン受容体拮抗薬、バソプレシン受容体作動薬、ムスカリン受容体拮抗薬、ムスカリン受容体作動薬、アドレナリン受容体拮抗薬、アドレナリン受容体作動薬、アンジオテンシン受容体作動薬、GABA受容体作動薬、トロンビン受容体拮抗薬、トロンビン受容体作動薬、オピオイド受容体作動薬、ADP受容体作動薬、ロイコトリエン受容体拮抗薬、ロイコトリエン受容体作動薬、メラトニン受容体作動薬、ソマトスタチン受容体作動薬、カンナビノイド受容体作動薬、スフィンゴシン1リン酸受容体作動薬、代謝型グルタミン酸受容体作動薬、ホスホリパーゼA2阻害剤、TGF−β産生抑制剤、Th2サイトカイン阻害剤およびそれらの2種以上の組合せから選択される第一の細胞性免疫誘導促進剤をテープ剤の形態で対象に経皮投与することを含む、細胞性免疫誘導方法;
(51)治療上有効量の(i)WT1ペプチドおよび/または改変WT1ペプチド、ならびに(ii)TLRリガンド、環状ジヌクレオチド、ヘルパーペプチド、免疫調節低分子薬物、シクロオキシゲナーゼ阻害剤、プロスタグランジン受容体拮抗薬、プロスタグランジン受容体作動薬、TSLP産生抑制剤、アデニル酸シクラーゼ阻害剤、オメガ3脂肪酸、PPAR作動薬、ドーパミン受容体拮抗薬、ドーパミン受容体作動薬、ヒスタミン受容体作動薬、ヒスタミン受容体拮抗薬、セロトニン受容体作動薬、セロトニン受容体拮抗薬、バソプレシン受容体拮抗薬、バソプレシン受容体作動薬、ムスカリン受容体拮抗薬、ムスカリン受容体作動薬、アドレナリン受容体拮抗薬、アドレナリン受容体作動薬、アンジオテンシン受容体作動薬、GABA受容体作動薬、トロンビン受容体拮抗薬、トロンビン受容体作動薬、オピオイド受容体作動薬、ADP受容体作動薬、ロイコトリエン受容体拮抗薬、ロイコトリエン受容体作動薬、メラトニン受容体作動薬、ソマトスタチン受容体作動薬、カンナビノイド受容体作動薬、スフィンゴシン1リン酸受容体作動薬、代謝型グルタミン酸受容体作動薬、ホスホリパーゼA2阻害剤、TGF−β産生抑制剤、Th2サイトカイン阻害剤およびそれらの2種以上の組合せから選択される第一の細胞性免疫誘導促進剤をテープ剤の形態で対象に経皮投与することを含む、癌の処置または予防方法;
(52)治療上有効量の(1)〜(49)のいずれかの癌ワクチンテープ剤を対象に経皮投与することを含む、癌の処置または予防方法;
(53)WT1ペプチドおよび/または改変WT1ペプチドの経皮投与による細胞性免疫誘導のために使用するための、(i)WT1ペプチドおよび/または改変WT1ペプチドと(ii)TLRリガンド、環状ジヌクレオチド、ヘルパーペプチド、免疫調節低分子薬物、シクロオキシゲナーゼ阻害剤、プロスタグランジン受容体拮抗薬、プロスタグランジン受容体作動薬、TSLP産生抑制剤、アデニル酸シクラーゼ阻害剤、オメガ3脂肪酸、PPAR作動薬、ドーパミン受容体拮抗薬、ドーパミン受容体作動薬、ヒスタミン受容体作動薬、ヒスタミン受容体拮抗薬、セロトニン受容体作動薬、セロトニン受容体拮抗薬、バソプレシン受容体拮抗薬、バソプレシン受容体作動薬、ムスカリン受容体拮抗薬、ムスカリン受容体作動薬、アドレナリン受容体拮抗薬、アドレナリン受容体作動薬、アンジオテンシン受容体作動薬、GABA受容体作動薬、トロンビン受容体拮抗薬、トロンビン受容体作動薬、オピオイド受容体作動薬、ADP受容体作動薬、ロイコトリエン受容体拮抗薬、ロイコトリエン受容体作動薬、メラトニン受容体作動薬、ソマトスタチン受容体作動薬、カンナビノイド受容体作動薬、スフィンゴシン1リン酸受容体作動薬、代謝型グルタミン酸受容体作動薬、ホスホリパーゼA2阻害剤、TGF−β産生抑制剤、Th2サイトカイン阻害剤およびそれらの2種以上の組合せから選択される第一の細胞性免疫誘導促進剤との組合せであって、テープ剤の形態で対象に経皮投与されるものである組合せ;
(54)癌の処置または予防に用いるための、(i)WT1ペプチドおよび/または改変WT1ペプチドと(ii)TLRリガンド、環状ジヌクレオチド、ヘルパーペプチド、免疫調節低分子薬物、シクロオキシゲナーゼ阻害剤、プロスタグランジン受容体拮抗薬、プロスタグランジン受容体作動薬、TSLP産生抑制剤、アデニル酸シクラーゼ阻害剤、オメガ3脂肪酸、PPAR作動薬、ドーパミン受容体拮抗薬、ドーパミン受容体作動薬、ヒスタミン受容体作動薬、ヒスタミン受容体拮抗薬、セロトニン受容体作動薬、セロトニン受容体拮抗薬、バソプレシン受容体拮抗薬、バソプレシン受容体作動薬、ムスカリン受容体拮抗薬、ムスカリン受容体作動薬、アドレナリン受容体拮抗薬、アドレナリン受容体作動薬、アンジオテンシン受容体作動薬、GABA受容体作動薬、トロンビン受容体拮抗薬、トロンビン受容体作動薬、オピオイド受容体作動薬、ADP受容体作動薬、ロイコトリエン受容体拮抗薬、ロイコトリエン受容体作動薬、メラトニン受容体作動薬、ソマトスタチン受容体作動薬、カンナビノイド受容体作動薬、スフィンゴシン1リン酸受容体作動薬、代謝型グルタミン酸受容体作動薬、ホスホリパーゼA2阻害剤、TGF−β産生抑制剤、Th2サイトカイン阻害剤およびそれらの2種以上の組合せから選択される第一の細胞性免疫誘導促進剤との組合せであって、テープ剤の形態で対象に経皮投与されるものである組合せ; および
(55)細胞性免疫誘導のための経皮投与用癌ワクチンテープ剤の製造における、(i)WT1ペプチドおよび/または改変WT1ペプチドならびに(ii)TLRリガンド、環状ジヌクレオチド、ヘルパーペプチド、免疫調節低分子薬物、シクロオキシゲナーゼ阻害剤、プロスタグランジン受容体拮抗薬、プロスタグランジン受容体作動薬、TSLP産生抑制剤、アデニル酸シクラーゼ阻害剤、オメガ3脂肪酸、PPAR作動薬、ドーパミン受容体拮抗薬、ドーパミン受容体作動薬、ヒスタミン受容体作動薬、ヒスタミン受容体拮抗薬、セロトニン受容体作動薬、セロトニン受容体拮抗薬、バソプレシン受容体拮抗薬、バソプレシン受容体作動薬、ムスカリン受容体拮抗薬、ムスカリン受容体作動薬、アドレナリン受容体拮抗薬、アドレナリン受容体作動薬、アンジオテンシン受容体作動薬、GABA受容体作動薬、トロンビン受容体拮抗薬、トロンビン受容体作動薬、オピオイド受容体作動薬、ADP受容体作動薬、ロイコトリエン受容体拮抗薬、ロイコトリエン受容体作動薬、メラトニン受容体作動薬、ソマトスタチン受容体作動薬、カンナビノイド受容体作動薬、スフィンゴシン1リン酸受容体作動薬、代謝型グルタミン酸受容体作動薬、ホスホリパーゼA2阻害剤、TGF−β産生抑制剤、Th2サイトカイン阻害剤およびそれらの2種以上の組合せから選択される第一の細胞性免疫誘導促進剤の使用。
本明細書において使用するとき、用語「WT1ペプチド」は、癌遺伝子WT1(Wilm’s腫瘍)の産物であるWT1蛋白が断片化された、約8〜約15個、好ましくは約8〜約12個のアミノ酸からなる部分ペプチドを意味し、Db126ペプチド、Db235抗原ペプチド等がこれに含まれる。また、WO−2000/06602にて開示されているWT1産物の部分ペプチド、WO2005/095598に記載のWT1由来HLA−A26結合性癌抗原ペプチド、WO2007/097358に記載のHLA−A*3303拘束性WT1ペプチド、およびWO2008/081701に記載のHLA−A*1101拘束性WT1ペプチドも本発明の「WT1ペプチド」に含まれる。
改変WT1ペプチドには、例えば、
(a)WT1ペプチドのアミノ酸配列において、1個から数個、例えば1個、2個、3個、4個または5個のアミノ酸が置換、欠失または付加されたアミノ酸配列からなるペプチド;および
(b)WT1ペプチドのアミノ酸配列において、全部または一部のアミノ酸、例えば1個または複数個、例えば1個、2個、3個、4個、5個、6個、7個、8個、9個、10個、11個または12個のアミノ酸が修飾されたアミノ酸配列からなるペプチド
が含まれる。
改変WT1ペプチドが有し得るアミノ酸の「修飾」としては、これらに限定されないが、例えば、アセチル化、メチル化などアルキル化、グリコシル化、ヒドロキシル化、カルボキシル化、アルデヒド化、リン酸化、スルホニル化、ホルミル化、ミリストイル化やパルミトイル化やステアロイル化のような脂肪鎖付加修飾、オクタノイル化、エステル化、アミド化、脱アミド化、シスチン修飾やグルタチオン修飾やチオグリコール酸修飾のようなジスルフィド結合形成修飾、糖化、ユビキチン化、スクシンイミド形成、グルタミル化、プレニル化等が挙げられる。改変WT1ペプチドは、1個以上のアミノ酸の置換、欠失または付加と、1個以上のアミノ酸の修飾を組み合わせて含むものであってもよい。
改変ヘルパーペプチドには、例えば、
(a)元のヘルパーペプチドのアミノ酸配列において、1個から数個、例えば1個、2個、3個、4個または5個のアミノ酸が置換、欠失または付加されたアミノ酸配列からなるペプチド;および
(b)元のヘルパーペプチドのアミノ酸配列において、全部または一部のアミノ酸、例えば1個または複数個、例えば1個、2個、3個、4個、5個、6個、7個、8個、9個、10個、11個、12個、13個、14個、15個、16個、17個、または18個のアミノ酸が修飾されたアミノ酸配列からなるペプチド
が含まれる。
改変ヘルパーペプチドが有し得るアミノ酸の「修飾」としては、これらに限定されないが、例えば、アセチル化、メチル化などのアルキル化、グリコシル化、ヒドロキシル化、カルボキシル化、アルデヒド化、リン酸化、スルホニル化、ホルミル化、ミリストイル化やパルミトイル化やステアロイル化のような脂肪鎖付加修飾、オクタノイル化、エステル化、アミド化、脱アミド化、シスチン修飾やグルタチオン修飾やチオグリコール酸修飾のようなジスルフィド結合形成修飾、糖化、ユビキチン化、スクシンイミド形成、グルタミル化、プレニル化等が挙げられる。また、改変ヘルパーペプチドは、1個以上のアミノ酸の置換、欠失または付加と、1個以上のアミノ酸の修飾を組み合わせて含むものであってもよい。
WT1ペプチドおよび/または改変WT1ペプチドが癌ワクチンとして有用なことは、既に明らかにされている(例えば特許文献1)。
さらに、この場合、取り扱い性の点から、25〜100μm程度の厚みを有するものが好ましい。
本明細書において使用するとき、本発明のテープ剤に含有させ得る「薬理学的に許容される塩」とは、投与対象に有害な作用を及ぼさず、かつ、該テープ剤中の成分の薬理活性を消失させない塩を意味する。無機酸塩(例えば塩酸塩やリン酸塩)、有機酸塩(例えば酢酸塩やフタル酸塩、TFA塩)、金属塩(アルカリ金属塩(例えば、ナトリウム塩、カリウム塩)、アルカリ土類金属塩(例えば、カルシウム塩、マグネシウム塩)、アルミニウム塩など)、アミン塩(トリエチルアミン塩、ベンジルアミン塩、ジエタノールアミン塩、t−ブチルアミン塩、ジシクロヘキシルアミン塩、アルギニン塩、ジメチルアンモニウム塩、アンモニウム塩など)を含むが、これらに限定されない。
かかる皮膚透過性増強剤としては例えば、オレイルアルコール、オクチルドデカノールなどの高級アルコール;グリセリン、エチレングリコール、ポリプロピレングリコールなどの多価アルコール;オレイン酸、カプリル酸などの高級脂肪酸;ミリスチン酸イソプロピル、パルミチン酸イソプロピル、オレイン酸エチルなどの脂肪酸エステル;セバシン酸ジエチル、アジピン酸ジイソプロピルなどの多塩基酸エステル;トリイソステアリン酸ジグリセリル、モノオレイン酸ソルビタン、ジカプリル酸プロピレングリコール、モノラウリン酸ポリエチレングリコール、テトラオレイン酸ポリオキシエチレンソルビットなどの多価アルコール脂肪酸エステル;ポリオキシエチレンラウリルエーテルなどのポリオキシエチレンアルキルエーテル;スクアラン、流動パラフィンなどの炭化水素;オリーブ油、ヒマシ油などの植物油;シリコーン油;N−メチルピロリドン、N−ドデシルピロリドンのようなピロリドン類;デシルメチルスルホキシドのようなスルホキシドなどが挙げられ、これらは1種で、または2種以上を混合して使用することができる。
テープ剤に用いる粘着剤を重合(アクリル系粘着剤A、B)、調合(PIBゴム系粘着剤、SIS系粘着剤A、SIS−PIB系粘着剤A)した。
(アクリル系粘着剤Aの重合)
不活性ガス雰囲気下、アクリル酸2−エチルへキシル75部、N−ビニル−2−ピロリドン22部、アクリル酸3部およびアゾビスイソブチロニトリル0.2部を酢酸エチル中60℃にて溶液重合させてアクリル系粘着剤A溶液を得た。
(アクリル系粘着剤Bの重合)
不活性ガス雰囲気下、アクリル酸2−エチルへキシル70部、N−ビニル−2−ピロリ
ドン25部、N−(2−ヒドロキシエチル)アクリルアミド5部およびアゾビスイソブチロニトリル0.2部を酢酸エチル中60℃にて溶液重合させてアクリル系粘着剤B溶液を得た。
(PIBゴム系粘着剤の調合)
ポリイソブチレン(オパノールB200、BASF社製)24部、ポリイソブチレン(オパノールB12、BASF社製)36部および脂環族系石油樹脂(アルコンP−100、荒川化学社製)40部をトルエンに溶解して、PIBゴム系粘着剤溶液を得た。
(SIS系粘着剤Aの調合)
スチレン-イソプレン-スチレンブロック共重合体(SIS5002、JSR社製)60部、および脂環族系石油樹脂(アルコンP−100、荒川化学社製)40部をトルエンに溶解して、SIS系粘着剤A溶液を得た。
(SIS−PIB系粘着剤Aの調合)
スチレン-イソプレン-スチレンブロック共重合体(SIS5002、JSR社製)30部、ポリイソブチレン(オパノールB100、BASF社製)30部および脂環族系石油樹脂(アルコンP−100、荒川化学社製)40部をトルエンに溶解して、SIS−PIB系粘着剤A溶液を得た。
上記の通りに製造したテープ剤について、免疫評価用モデル動物を用いてマウス免疫試験を行った。免疫誘導レベルの評価は、ELISPOT法によって行った。具体的には、マウス背部を毛刈りし、毛刈りによる皮膚ダメージを回復させるための飼育期間を設けた後、マウスの背部皮膚にサンプルを所定時間投与して除去し、所定日数の飼育を行い、抗原特異的な細胞性免疫の誘導レベルを評価した。投与から所定日数経過後に脾臓を摘出し、脾細胞懸濁液を調製した。抗マウスIFN−γ抗体を固定化したELISPOTプレートのウェルに、脾細胞(3×106cells/well)と抗原ペプチド(100μM)とを培養液とともに入れ、37℃、5%CO2の培養条件にて20時間、共培養し、ELISPOT法にてIFN−γ産生細胞スポット数(スポット数/3×106cells)を評価した。
テープ剤の投与量は上記の通り0.7cm2、投与回数は(24hr/週)×1回、脾臓摘出は投与から6日後とした。用いたマウスは、HLA−A*0201型MHC拘束性ペプチドによる細胞性免疫誘導を評価可能な遺伝子改変マウスである。
皮膚刺激評価用モデル動物としてのC57BL/6マウスを用いてTSLPレベルを評価した。免疫評価用モデル動物に対する投与条件と同じ条件にて、皮膚刺激評価用モデル動物に対して製剤投与を行い、製剤投与終了時に、マウス背部皮膚を摘出し、抽出溶媒中(プロテアーゼ阻害剤(Protease Inhibitor Cocktail for general use,SIGMA―ALDRICH製)と10μMインドメタシン(和光純薬製)を含むPBS溶液)でホモジナイザー(ヒスコトロン、マイクロテック・ニチオン製)を用い皮膚を破砕した。破砕した皮膚を4℃、9000gで10分間遠心分離した後、上清を回収した。上清中のTSLP量をELISA(Mouse TSLP Quantikine ELISA Kit、R&D Systems製)により測定した。また、上清中の総タンパク量をBCA法(Pierce BCA Protein Assay Kit、Thermo SCIENTIFIC)により測定し、TSLP量を総タンパク量で除することで標準化した。
携帯型閉鎖チャンバー方式水分蒸散量測定装置(アサヒバイオメッド製、VAPO SCAN AS−VT100RS)を用い、マウス皮膚に当該機器を5〜15秒程度接触させる事により測定を行った。マウス皮膚に前処理を行った10分後に測定した値を経皮水分蒸散量(TEWL)(g/h・m2)とした。
フランツ型拡散セルを用いて、Db126ペプチドならびにイミキモドの皮膚透過試験を行った。予め毛刈りをしたマウスの背部より摘出した皮膚を、37℃のリン酸バッファー(pH7.4等張緩衝液)を循環させたフランツ型拡散セル(適用面積4.91cm2)に装着した。該装着した皮膚上に0.7cm2の製剤を貼付し、24時間後にセル内の試料を採取した。採取した試料は、高速液体クロマトグラフ・タンデム型質量分析計に供し、予め定めておいた検量線より、24時間後の皮膚を透過したDb126ペプチドの量(Db126ペプチド透過量、μg/cm2/24hr)ならびにイミキモドの量(イミキモド透過量、μg/cm2/24hr)を算出した。
最終免疫投与から7日後に下記の要領で脾臓細胞(ターゲット細胞あるいはコントロール細胞)を移殖し、その18時間後に脾臓を採取し、FACS測定を行うことにより % 特異的溶解を求めた。
ナイーブマウス(免疫試験に用いたのと同じ種類の免疫評価用モデル動物)から脾臓を摘出し、RPMI1640培地を入れたシャーレ中でスライドガラスを用いてすりつぶした。50 mLチューブに回収後、10℃、1100rpm、5分遠心分離した後、上清を捨て、Lysis Buffer 20 mLを加え、室温で5分インキュベートした。培地を20 mL加えた後、遠心分離した後、培地を加え、セルストレーナーに通した。
手順1で調製した脾臓細胞を10℃、1100rpm、5分遠心分離した後、上清を捨て、HBSS bufferを加えて2×107 cells/mLとした。この細胞液を2本の50mLチューブに分注し、一方の細胞液に最終濃度10μMとなるように100μM抗原溶液(抗原は各免疫投与物に配合した抗原)を加えてターゲット細胞とした。もう一方の細胞はコントロール細胞とした。両方の細胞を37℃で1時間インキュベートした後、遠心分離し、上清を捨て、培地を加えた。
手順2で抗原標識した細胞を遠心分離し、0.1% BSA-PBSを1×107 cells/mLとなるように加えた。ターゲット細胞液に5 mM CFSE溶液を最終濃度10μMとなるように加え、コントロール細胞液に5 mM CFSE溶液を最終濃度1μMとなるように加え、ボルテックスした後37℃で10分インキュベートした。その後遠心分離し、上清を捨て、培地を加えた。
手順3でCFSE標識した細胞を遠心分離し、上清を捨て、HBSS bufferを用い、5×107 cells/mLとなるように調製した。ターゲット細胞液とコントロール細胞液を等量混合し、免疫したマウスに200 μLずつ眼窩静脈から投与した(移植細胞数:1×107 cells/匹)。
脾臓細胞移植の18時間後、脾臓を摘出し、手順1と同様に脾臓細胞を調製した。その後、FACSによりCFSE陽性細胞を検出し、CFSE high細胞(ターゲット細胞)とCFSE low細胞(コントロール細胞)の比率から、以下の式より細胞傷害活性を評価した。この指標により抗原特異的に誘導した免疫が生体内で抗原提示細胞を特異的に攻撃する能力を評価し、本発明の投与物の効果が高いことを確認した。
r= (% CFSE low 細胞)/(% CFSE high 細胞)
% 特異的溶解=(1-(r非免疫/r免疫))×100
下記表4の組成を有する皮内注射剤を製造した。具体的には、表4に記載される配合量のDb126ペプチドおよびアジュバントとしてのMontanide ISA51VG(フロイント産業)に、基剤としての生理食塩水を添加して合計100重量部とし、ホモジナイザーにて混和してエマルジョン状の注射剤を調製した。
上記の通りに製造した注射剤を用いて、上記マウス免疫試験1と同様の方法によりマウス免疫試験を行った。注射剤をマウスの背部に皮内注射投与した。投与量は30μL、投与回数は1回、脾臓摘出は投与から6日後とした。使用したマウスはHLA−A*0201型MHC拘束性ペプチドによる細胞性免疫誘導を評価可能な遺伝子改変マウスである。免疫試験の結果を下記表4に示す。
好ましくは、TLRリガンド、環状ジヌクレオチド、シクロオキシゲナーゼ阻害剤、TSLP産生抑制剤、プロスタグランジン受容体拮抗薬、プロスタグランジン受容体作動薬、PPAR作動薬、TGF-β産生抑制剤、ヒスタミン受容体作動薬、ヒスタミン受容体拮抗薬、セロトニン受容体作動薬、セロトニン受容体拮抗薬、ムスカリン受容体拮抗薬、アドレナリン受容体拮抗薬、オピオイド受容体作動薬、メラトニン受容体作動薬、代謝型グルタミン酸受容体作動薬およびそれらの2種以上の組合せから選択される第一の細胞性免疫誘導促進剤、並びにヘルパーペプチド以外のそれらの第一の細胞性免疫誘導促進剤とヘルパーペプチドとの組合せが有効であった。
さらに好ましくは、TLR7および/またはTLR8リガンド、環状ジヌクレオチド、シクロオキシゲナーゼ阻害剤、TSLP産生抑制剤、プロスタグランジン受容体拮抗薬、プロスタグランジン受容体作動薬、PPAR作動薬、ヒスタミン受容体拮抗薬、セロトニン受容体作動薬、オピオイド受容体作動薬、メラトニン受容体作動薬、およびそれらの2種以上の組合せから選択される第一の細胞性免疫誘導促進剤、または並びにヘルパーペプチド以外のそれらの第一の細胞性免疫誘導促進剤とヘルパーペプチドとの組合せが特に有効であった。
Claims (11)
- (i)WT1ペプチドおよび/または改変WT1ペプチド;ならびに
(ii)TLRリガンド、環状ジヌクレオチド、ヘルパーペプチド、免疫調節低分子薬物、シクロオキシゲナーゼ阻害剤、プロスタグランジン受容体拮抗薬、プロスタグランジン受容体作動薬、TSLP産生抑制剤、アデニル酸シクラーゼ阻害剤、オメガ3脂肪酸、PPAR作動薬、ドーパミン受容体拮抗薬、ドーパミン受容体作動薬、ヒスタミン受容体作動薬、ヒスタミン受容体拮抗薬、セロトニン受容体作動薬、セロトニン受容体拮抗薬、バソプレシン受容体拮抗薬、バソプレシン受容体作動薬、ムスカリン受容体拮抗薬、ムスカリン受容体作動薬、アドレナリン受容体拮抗薬、アドレナリン受容体作動薬、アンジオテンシン受容体作動薬、GABA受容体作動薬、トロンビン受容体拮抗薬、トロンビン受容体作動薬、オピオイド受容体作動薬、ADP受容体作動薬、ロイコトリエン受容体拮抗薬、ロイコトリエン受容体作動薬、メラトニン受容体作動薬、ソマトスタチン受容体作動薬、カンナビノイド受容体作動薬、スフィンゴシン1リン酸受容体作動薬、代謝型グルタミン酸受容体作動薬、ホスホリパーゼA2阻害剤、TGF−β産生抑制剤、Th2サイトカイン阻害剤、およびそれらの2種以上の組合せから選択される第一の細胞性免疫誘導促進剤
を含む、細胞性免疫誘導のための経皮投与用癌ワクチンテープ剤。 - さらに第二の細胞性免疫誘導促進剤としての薬理学的に許容される酸またはその薬理学的に許容される塩を含む、請求項1の経皮投与用癌ワクチンテープ剤。
- 第一の細胞性免疫誘導促進剤がヘルパーペプチドである、請求項1または2の経皮投与用癌ワクチンテープ剤。
- 第一の細胞性免疫誘導促進剤が、TLRリガンド、環状ジヌクレオチド、免疫調節低分子薬物、シクロオキシゲナーゼ阻害剤、プロスタグランジン受容体拮抗薬、プロスタグランジン受容体作動薬、TSLP産生抑制剤、アデニル酸シクラーゼ阻害剤、オメガ3脂肪酸、PPAR作動薬、ドーパミン受容体拮抗薬、ドーパミン受容体作動薬、ヒスタミン受容体作動薬、ヒスタミン受容体拮抗薬、セロトニン受容体作動薬、セロトニン受容体拮抗薬、バソプレシン受容体拮抗薬、バソプレシン受容体作動薬、ムスカリン受容体拮抗薬、ムスカリン受容体作動薬、アドレナリン受容体拮抗薬、アドレナリン受容体作動薬、アンジオテンシン受容体作動薬、GABA受容体作動薬、トロンビン受容体拮抗薬、トロンビン受容体作動薬、オピオイド受容体作動薬、ADP受容体作動薬、ロイコトリエン受容体拮抗薬、ロイコトリエン受容体作動薬、メラトニン受容体作動薬、ソマトスタチン受容体作動薬、カンナビノイド受容体作動薬、スフィンゴシン1リン酸受容体作動薬、代謝型グルタミン酸受容体作動薬、ホスホリパーゼA2阻害剤、TGF−β産生抑制剤、およびTh2サイトカイン阻害剤からなる群より選択される1種以上とヘルパーペプチドとの組み合わせである、請求項1または2の経皮投与用癌ワクチンテープ剤。
- テープ剤の粘着剤がアクリル系粘着剤である、請求項1〜4のいずれか1項の経皮投与用癌ワクチンテープ剤。
- テープ剤の粘着剤がゴム系粘着剤である、請求項1〜4のいずれか1項の経皮投与用癌ワクチンテープ剤。
- テープ剤の粘着剤がシリコーン系粘着剤である、請求項1〜4のいずれか1項の経皮投与用癌ワクチンテープ剤。
- テープ剤の粘着剤層がさらに皮膚透過性増強剤を含む、請求項5〜7のいずれか1項の経皮投与用癌ワクチンテープ剤。
- 低刺激条件下で投与されるものである、請求項1〜8のいずれか1項の経皮投与用癌ワクチンテープ剤。
- 低刺激条件が、皮膚刺激評価用モデル動物における該テープ剤の投与前の経皮水分蒸散量(TEWL)が50g/h・m2以下の条件である、請求項9の経皮投与用癌ワクチンテープ剤。
- 低刺激条件が、皮膚刺激評価用モデル動物における該テープ剤の投与終了時の皮膚内TSLPレベルが10000pg/mgタンパク質以下の条件である、請求項9または10の経皮投与用癌ワクチンテープ剤。
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- 2014-01-29 CN CN201410042748.8A patent/CN103961305B/zh not_active Expired - Fee Related
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Also Published As
Publication number | Publication date |
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US10195258B2 (en) | 2019-02-05 |
RU2014102937A (ru) | 2015-08-10 |
CA2841014A1 (en) | 2014-08-05 |
US20140220105A1 (en) | 2014-08-07 |
CN103961305B (zh) | 2019-10-18 |
CN103961305A (zh) | 2014-08-06 |
KR102050931B1 (ko) | 2019-12-02 |
EP2762156A1 (en) | 2014-08-06 |
RU2697443C2 (ru) | 2019-08-14 |
IN2014CH00393A (ja) | 2015-04-03 |
JP6512569B2 (ja) | 2019-05-15 |
KR20140100421A (ko) | 2014-08-14 |
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