JP2019127486A - 経口用抗アレルギー剤 - Google Patents
経口用抗アレルギー剤 Download PDFInfo
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- JP2019127486A JP2019127486A JP2019008551A JP2019008551A JP2019127486A JP 2019127486 A JP2019127486 A JP 2019127486A JP 2019008551 A JP2019008551 A JP 2019008551A JP 2019008551 A JP2019008551 A JP 2019008551A JP 2019127486 A JP2019127486 A JP 2019127486A
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- Prior art keywords
- fucoxanthin
- treg
- derivative
- salt
- lipopolysaccharide
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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Abstract
Description
項2.リポ多糖及び/又はTLR2アゴニスト活性を有する多糖類を更に含む、項1に記載の抗アレルギー剤。
項3.前記アレルギーが、I型アレルギー及び/又はIV型アレルギーである、項1又は2に記載の抗アレルギー剤。
項4.前記アレルギーが、アレルギー性鼻炎、花粉症、アトピー性皮膚炎、アレルギー性結膜炎、アレルギー性気管支喘息、食物アレルギー、蕁麻疹、及び接触性皮膚炎からなる群から選択される少なくとも1種である、項1〜3のいずれか一項に記載の抗アレルギー剤。
項5.フコキサンチン若しくはその誘導体又はその塩を有効成分とするTh2抑制及びTreg活性化剤。
項6.リポ多糖及び/又はTLR2アゴニスト活性を有する多糖類を更に含む、項5に記載のTh2抑制及びTreg活性化剤。
項7.フコキサンチン若しくはその誘導体又はその塩を有効成分とするGATA3発現抑制及びFoxP3発現促進剤。
項8.フコキサンチン若しくはその誘導体又はその塩、並びに/又はリポ多糖及び/若しくはTLR2アゴニスト活性を有する多糖類を有効成分とするGATA3発現抑制用外用剤。
ファイトロックス社製、Infini-Pureフコキサンチン40を、シリカゲルクロマトグラフィーにより分離し、Journal of Pharmacological Sciences Volume 132, Issue 1, 2016, Pages 55-64に記載のある通りに精製した。
フコキサンチン処理によるGATA3発現への影響をリアルタイムPCRにより検証した。
GATA-1-F 5'-CGCTCCCTGTCACCGGCAGTGC-3'(AN: NM_008089)(配列番号1)
GATA-1-R 5'-CCGCCACAGTGGAGTAGCCGTT-3'(配列番号2)
GATA-2-F 5'-CTCCCGACGAGGTGGATGTCTT-3'(AN: NM_008090)(配列番号3)
GATA-2-R 5'-CCTGGGCTGTGCAACAAGTGTG-3'(配列番号4)
GATA-3-F 5'-TTTACCCTCCGGCTTCATCCTCTT-3'(AN: NM_008091)(配列番号5)
GATA-3-R 5'-TGCACCTGATACTTGAGGCACTCT-3'(配列番号6)
初代CD4+T細胞は、CD4+細胞単離キット及びLS Column (Miltenyi Biotec, Bergisch Gladbach, Germany)を用いてBL6雄マウスの脾臓から単離した。それから、細胞はCD3/CD28 T細胞活性化ビーズ(Life Technologies, Tokyo, Japan)と共に10μMのフコキサンチンの存在下又は非存在下で24時間インキュベートした。細胞は遠心により回収し、試験例1と同様に、qPCR解析を行った。
mIL-4-F 5'-CATCGGCATTTTGAACGAG-3'(AN: NM_021283)(配列番号7)
mIL-4-R 5'-CGAGCTCACTCTCTGTGGTG-3'(配列番号8)
mIL-5-F 5'-TTGACAAGCAATGAGACGATGAG-3'(AN: NM_010558)(配列番号9)
mIL-5-R 5'-GCCCCTGAAAGATTTCTCCAA-3'(配列番号10)
mIL-13-F 5'-GTGCCAAGATCTGTGTCTCTCC-3'(AN: NM_008355)(配列番号11)
mIL-13-R 5'-TTACAGAGGCCATGCAATATCC-3'(配列番号12)
初代CD4+T細胞は、試験例2と同様に調製し、CD3/CD28 T細胞活性化ビーズと共に10μMのフコキサンチンの存在下又は非存在下で24時間インキュベートした。インキュベート後、細胞は中性緩衝ホルマリンを用いて10分間固定化し、各抗体と反応させた。次に、FACS分析を行った(MACSQuant Analyzer 10, Miltenyi Biotech)。
FACSにて、T細胞サブセットのマーカーを指標に、免疫染色後、解析した。すべて、ミルテニー社の抗体を用いた。細胞は、試験例3と同様に調製した。
ラテックスビーズ(カルボキシル基修飾ポリスチレン製、蛍光レッド、平均粒子径2μm)(Merck, Darmstadt, Germany)を用いて、マクロファージ細胞の貧食作用に対するフコキサンチンの効果を試験した。細胞は、10 ng/mlのDer f1の存在下又は非存在下で、10μMのフコキサンチンを添加又は添加せず24時間インキュベートした。
10週齢の雄Nc/Ngaマウスに6日間3 mg/kgの量でLPSを経口投与した。1%カルボキシメチルセルロース溶液(CMC)に懸濁したフコキサンチンを1 mg/kgでマウスに1日1回経口投与した。全ての対照群はCMCを投与した。6日間投与後、マウスを安楽死させ、脾臓を摘出した。脾臓は、1 mlの血清不含RPMI 1640培地(Nakarai Tesque, Kyoto, Japan)に懸濁し、ナイロンメッシュ(70μm)でろ過した後、10 mlの懸濁液とした。細胞濃度を1.0×107 cellsをそれぞれの適切な抗体で染色した。FoxP3/CD4ダブルポジティブの細胞をFACSで測定した。
ヒトケラチノサイトHaCat細胞にダニアレルゲンDer f1 10 ng/mlを適用し、1μMのフコキサンチン及び10 ng/ml LPSの存在下又は非存在下で4日間インキュベートした。細胞は遠心により回収し、試験例1と同様に、qPCR解析を行った。
hGATA3-F 5'-GCTGTCTGCAGCCAGGAGAGC-3' (配列番号13)
hGATA3-R 5'-ATGCATCAAACAACTGTGGCCA-3' (配列番号14)
Claims (7)
- フコキサンチン若しくはその誘導体又はその塩を有効成分とする経口用抗アレルギー剤。
- リポ多糖及び/又はTLR2アゴニスト活性を有する多糖類を更に含む、請求項1に記載の抗アレルギー剤。
- 前記アレルギーが、I型アレルギー及び/又はIV型アレルギーである、請求項1又は2に記載の抗アレルギー剤。
- 前記アレルギーが、アレルギー性鼻炎、花粉症、アトピー性皮膚炎、アレルギー性結膜炎、アレルギー性気管支喘息、食物アレルギー、蕁麻疹、及び接触性皮膚炎からなる群から選択される少なくとも1種である、請求項1〜3のいずれか一項に記載の抗アレルギー剤。
- フコキサンチン若しくはその誘導体又はその塩を有効成分とするTh2抑制及びTreg活性化剤。
- リポ多糖及び/又はTLR2アゴニスト活性を有する多糖類を更に含む、請求項5に記載のTh2抑制及びTreg活性化剤。
- フコキサンチン若しくはその誘導体又はその塩、並びに/又はリポ多糖及び/若しくはTLR2アゴニスト活性を有する多糖類を有効成分とする、GATA3発現抑制用外用剤。
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JP2007084518A (ja) * | 2005-09-26 | 2007-04-05 | Kochi Univ | 海藻由来の免疫抑制物質を製造する方法 |
JP2009108022A (ja) * | 2008-05-01 | 2009-05-21 | Tsujido Kagaku Kk | 治療剤 |
JP2012254959A (ja) * | 2011-06-09 | 2012-12-27 | Kikkoman Corp | Il−17産生抑制剤 |
JP2013129627A (ja) * | 2011-12-21 | 2013-07-04 | Hydrox Kk | フロロタンニン類の簡易な調製方法とその利用 |
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JP2007084518A (ja) * | 2005-09-26 | 2007-04-05 | Kochi Univ | 海藻由来の免疫抑制物質を製造する方法 |
JP2009108022A (ja) * | 2008-05-01 | 2009-05-21 | Tsujido Kagaku Kk | 治療剤 |
JP2012254959A (ja) * | 2011-06-09 | 2012-12-27 | Kikkoman Corp | Il−17産生抑制剤 |
JP2013129627A (ja) * | 2011-12-21 | 2013-07-04 | Hydrox Kk | フロロタンニン類の簡易な調製方法とその利用 |
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