JP3858080B2 - アレルゲン分解剤、および抗アレルゲン羽毛 - Google Patents
アレルゲン分解剤、および抗アレルゲン羽毛 Download PDFInfo
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- JP3858080B2 JP3858080B2 JP2005515390A JP2005515390A JP3858080B2 JP 3858080 B2 JP3858080 B2 JP 3858080B2 JP 2005515390 A JP2005515390 A JP 2005515390A JP 2005515390 A JP2005515390 A JP 2005515390A JP 3858080 B2 JP3858080 B2 JP 3858080B2
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- Prior art keywords
- metal phthalocyanine
- formula
- allergen
- feather
- salt
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- LRBQNJMCXXYXIU-NRMVVENXSA-N tannic acid Chemical compound OC1=C(O)C(O)=CC(C(=O)OC=2C(=C(O)C=C(C=2)C(=O)OC[C@@H]2[C@H]([C@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)O2)OC(=O)C=2C=C(OC(=O)C=3C=C(O)C(O)=C(O)C=3)C(O)=C(O)C=2)O)=C1 LRBQNJMCXXYXIU-NRMVVENXSA-N 0.000 description 1
- 229940033123 tannic acid Drugs 0.000 description 1
- 235000015523 tannic acid Nutrition 0.000 description 1
- 229920002258 tannic acid Polymers 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 230000002110 toxicologic effect Effects 0.000 description 1
- 231100000027 toxicology Toxicity 0.000 description 1
- 230000001052 transient effect Effects 0.000 description 1
- 238000002562 urinalysis Methods 0.000 description 1
- 101150005573 uvrA gene Proteins 0.000 description 1
- 230000008673 vomiting Effects 0.000 description 1
- 239000002023 wood Substances 0.000 description 1
- RKQOSDAEEGPRER-UHFFFAOYSA-L zinc diethyldithiocarbamate Chemical compound [Zn+2].CCN(CC)C([S-])=S.CCN(CC)C([S-])=S RKQOSDAEEGPRER-UHFFFAOYSA-L 0.000 description 1
- 150000003754 zirconium Chemical class 0.000 description 1
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Description
本発明のアレルゲン分解剤の有効性を確かめるため、金属フタロシアニンの誘導体として鉄フタロシアニンテトラカルボン酸につき、試験管実験による薬効の確認と動物実験、その他の実験による安全性の確認をおこなった。
アレルゲンとしてダニ抗原DerfII(アサヒビール株式会社製)を選び、このダニアレルゲンを鉄フタロシアニンテトラカルボン酸と混合した場合の電気泳動と、アレルゲンのみの電気泳動を比較した。
各実験調製例の所定濃度(w/v%)の鉄フタロシアニンテトラカルボン酸カリウム(Fe−Pc−COOK)水溶液およびDerfII溶液と、IPG(Immobilized pH Gradient: 固定化pH勾配)バッファーを含んだ膨潤用ストック溶液(Urea 8mol/L、CHAPS 2w/v%、IPG Buffer 2%、Bromophenol blue 適量、蒸留水)との混合溶液を調製し、表4の実験調製例1〜4の試験溶液とした。
一次元電気泳動用ゲルのフィルム(以下Stripという)を2種類、Strip pH4〜7およびStrip pH3〜10.5を用意し、各試験溶液に浸漬して乾燥防止のシリコンオイルを適量添加し、10時間静置した。試験溶液から各Stripを取り出して水洗し、一次元電気泳動装置にセットし、乾燥防止のシリコンオイルを添加し、一次元電気泳動を16時間行った。
一次元電気泳動の終了したStripを泳動装置より取り出し水洗した。SDS(sodium dodecyl sulfate:ドデシル硫酸ナトリウム)平衡化用バッファー(1.5mol/L pH8.8 Tris−Cl 50mmol/L、Urea 6mol/L、87v/v% Glycerol 30v/v%、SDS 2v/v%、Bromophenol blue 適量、蒸留水)にDTT(dithiothreitol:ジチオトレイトール)100mg/10mLを加えた溶液10mLにStripを浸漬し10分間浸透させた。Strip を取り出し、SDS平衡化用バッファーにiodoacetamide(250mg/10mL)を加えた溶液10mLに浸漬し10分間浸透させた。Strip を取り出して水洗しふき取った後、二次元電気泳動ゲル、ろ紙、電極ゲル、分子量マーカーをセットした二次元電気泳動装置に取り付け、1時間40分電気泳動を行った。
資料の作成には、Amersham Biosciences製のSilver Staining Kit, Proteinを使用した。
本発明のアレルゲンの分解剤が、安全に使用できることを実証するために、鉄フタロシアニンテトラカルボン酸について以下の実験を行った。
鉄フタロシアニンテトラカルボン酸を1重量%含有するニット生地を、アレルゲン分解の機能性繊維の試料とした。
前記同一のアレルゲン分解の機能性繊維を試料とし、同様にウサギの損傷皮膚とした。正常皮膚と損傷皮膚の各1ヵ所に注射用水で湿らせた機能性繊維を貼付し、ガーゼを被せてからテーピングテープで固定した。他の各1ヵ所は無処置部位とし、ガーゼとテーピングテープの貼付を同様に行った。さらに布製カバー及びチューブ型ネット包帯を用いて被覆固定した。23時間後に機能性繊維、ガーゼ及びテーピングテープを除去し、投与部位を微温湯にて清拭した。投与期間は21日間とし、毎日の除去後1時間に刺激性を肉眼的に評価した。判定はDraizeの判定基準に準拠して、紅斑(痂皮形成)及び浮種についてそれぞれ評価した。正常皮膚と損傷皮膚に分けて、各観察時点ごとに紅斑(痂皮形成)、浮種及びそれらの合計評点(TS)について平均値と標準偏差を算出した。投与期間終了後に動物をペントバルビタールナトリウム麻酔下で放血致死させ、各投与部位皮膚を10%中性緩衝ホルマリン液で固定後、HE染色を施し病理組織学的検査を行った。
10週齢の日本白色種雄性ウサギ各3匹からなる非洗浄群及び洗浄群の2群を設定した。ウサギは7日間の予備飼育の後、体重推移及び一般状態に異常のないことを確認した。また、投与前日に両眼について肉眼的に異常のないこと、フルオレセイン染色(フルオル試験紙、ロット番号3990849、ワイス・アイアースト ラボラトリーズ、アメリカ)を施し、スリットランプ(SL−5型、興和(株))を用いて角膜に損傷のないことを確認して試験に供した。
塩化ナトリウム844mg、塩化カリウム1200mg、塩化カルシウム146mg、塩化マグネシウム52mg、リン酸二カリウム342mg、精製水1000mLで調製した生理食塩液に、被験物質である機能性繊維を0.2g/lmLの割合で浸し、オートクレーブ内で121±2℃で1時間抽出した後、20〜30℃に冷却し室温で保存し24時間以内に使用した。
5週齢のCrj:CD(SD)IGS(SPF)ラット雌雄各26匹を1週間予備検疫飼育し、異常は認められなかったことから全数を試験に供した。予備飼育前に測定した体重を基準として層物連続無作為化法により各群に割り付けた。群に割り付けられなかった余剰動物は群分けした。投与時の週齢は雌雄とも6週齢、体重は雄が178〜197g、雌が122〜142gであった。
5週齢のCrj:CD(SD)IGS(SPF)ラット雌雄各26匹を1週間検疫飼育し、異常が認められなかったことから全数を試験に供した。検疫飼育の直前に測定した体重を基準として層物連続無作為化法により各群に割り付けた。群に割り付けられなかった余剰動物は群分け実施日に試験から除外し、安楽死させ処分した。投与時の週齢は雌雄とも6週齢、体重は雄が172〜186g、雌が130〜151gであった。
5週齢の雄性Crj;Hartley系モルモットを媒体対照群、被験物質感作群に分け、Maximization法により機能性繊維の皮膚感作性について試験した。被験物質として細切した機能性繊維に10倍量のメタノールを加えて室温にて抽出した後、メタノールを留去し、機能性繊維151.88gから5.482gの抽出物が得られた。
7の皮膚感作性試験に使用した機能性繊維メタノール抽出物について、5週齢の雄性Crj:Hartley系モルモットを用い、Adjuvant and Strip法により皮膚光感作性試験をした。各投与物質0.1mLを除毛したモルモット背部に開放塗布し、30分後より約10.2Joules/cm2の紫外線を照射した。同様の処置を5日間連続して行い、光感作した。最終感光感作後17日に各投与物質0.1mLを除毛した背部皮膚の左右対称に開放塗布し、動物の右半分をアルミホイルで被覆して約10.2Joules/cm2の紫外線を照射して光惹起した。光惹起後24時間及び48時間に皮膚反応を判定した。
7の皮膚感作性試験に使用した機能性繊維メタノール抽出物について、5週齢の雄性Crj:Htley系モルモットに対し、Morikawaらの方法により光毒性試験をした。被験物質投与群については10%の濃度でジメチルスルホキシドに溶解した機能性繊維のメタノール抽出物を、媒体対照群についてはジメチルスルホキシドを、陽性対照群については0.05% 8−methoxypsoralenを、それぞれ投与物質として、除毛したモルモットの背部に各投与物質0.03mLを開放塗布した。開放塗布後30分に約11.2Joules/cm2の紫外線を照射した。照射後24及び48時間に皮膚反応を判定した。
ネズミチフス菌Salmonella typhimurium TA100、TA1535、TA98、TA1537及び大腸菌Escherichia coli WP2 uvrAを使用して、機能性繊維のメタノール抽出物の突然変異誘発能の有無を検索した。メタノール抽出物は、細切した機能性繊維に10倍量のメタノールを加え、室温で24時間攪拌して抽出し、ロータリーエバポレーターでメタノールを留去した残留物である。
機能性繊維のメタノール抽出物の染色体異常誘発性の有無を検索した。DMSOを媒体として試験を実施した。細胞増殖抑制試験における被験物質の最高試験用量は、5.0mg/mLとした。細胞増殖抑制試験の結果、50%細胞増殖抑制濃度は、短時間処理の代謝活性化系非存在下(以下−S9mix法と略す)で、5.0mg/mL以上となった。また、短時間処理代謝活性化系存在下(以下+S9mix法と略す)で、0.840mg/mLとなった。被験物質の沈澱が、0.313mg/mL以上で認められた。したがって、染色体異常試験の試験用量は、−S9mix法の最高試験用量を0.313mg/mLとし、公比2で希釈した3用量を設定した。また、+S9mix法の最高試験用量を1.25mg/mLとし、公比2で希釈した4用量を設定した。
染色分体型切断(ctb)
染色分体型交換(四放射状交換など、cte)
染色体型切断(csb)
染色体型交換(ニ動原体、環状など、cse)
その他(断片化、frg)
その他の染色体異常
ギャップ(g)
数的異常
倍数性(polyploid、endoreduplication)
被験物質である機能性繊維8gを、高圧蒸気滅菌(121℃、20分)した。冷却、乾燥後、培地80mLを加えて軽く栓をした後、被験物質が培地中に十分に浸漬していることを確認して、炭酸ガスインキュベーター内に静置して24時間インキュベートした。ガラス瓶から培地のみを取り出し、この培地を100%被験物質抽出液とした。培地を用いて100、80、70、50、30及び10%に希釈した。なお、上記の被験物質抽出液の濃度は、予備検討の結果からIC50値を算出できる適切な範囲で6濃度設定した。
前除塵処理、洗浄前除塵処理および水での洗浄処理を行ったグース羽毛(ダウン85質量%、スモールフェザー15質量%)(かさ高性153mm)を準備した。
コバルトフタロシアニンポリスルホン酸ナトリウムは大部分、羽毛に吸尽された。
前除塵処理、洗浄前除塵処理および水での洗浄処理を行ったグース羽毛(ダウン85質量%、スモールフェザー15質量%)を準備した。
1.ダニアレルゲン吸着力試験
実施例2および3で得た、金属フタロシアニン化合物(II)またはその塩が担持された抗アレルゲン羽毛を試料とし、ダニアレルゲンの吸着力を以下のようにして測定した。対照試料としては、実施例2および3において、試薬溶液処理を行っていない羽毛、すなわち、前除塵処理、洗浄前除塵処理および水での洗浄処理を行ったグース羽毛(ダウン85質量%、スモールフェザー15質量%)を用いた。
(1)抗原のコーティング
マイクロプレート(塩化ビニル製96ウエルプレート、Dynatech社製)に、前記上澄み液を100μl/ウエル注入し、25℃で2時間維持した。その後、マイクロピペットを用いて、前記上澄み液をウエルから除去した。PBS溶液で3回ウエルを洗浄した。
BSA溶液(VECTOR 社製、BOVINESERUMALBUMIN(商品名))(1%(w/v))を、前記マイクロプレートに100μl/ウエル注入し、25℃で1時間維持した。その後、マイクロピペットを用いて、前記上澄み液をウエルから除去した。0.05%Tween−PBS溶液で1回ウエルを洗浄した。
前記マイクロプレートにダニアレルゲン抗体溶液(アサヒビール社製、抗Derf2モノクローナル抗体(商品名))(5μg/mL)を、100μl/ウエル注入し、25℃で2時間維持した。
ビオチン(Biotin)標識された抗マウスIgG(H+L)抗体溶液(VECTOR社製、BIOTINYLATED ANTI−MOUSE IgG(H+L)(商品名))(1μg/mL)を、前記マイクロプレートに100μl/ウエル注入し、25℃で2時間維持した。その後、マイクロピペットを用いて、前記抗体溶液をウエルから除去した。0.05%Tween−PBS溶液で1回ウエルを洗浄した。
アビジン溶液(VECTOR社製、ALKALINE PHOSPHATASE AVIDIN D(商品名))(100unit)を、前記マイクロプレートに100μl/ウエル注入し、25℃で0.5時間維持した。その後、マイクロピペットを用いて、前記上澄み液をウエルから除去した。0.05%Tween−PBS溶液で3回ウエルを洗浄した。
PNPP溶液(VECTOR社製、P−NITROPHENYL PHOSPHATE(商品名))(500μg/mL)を、前記マイクロプレートに100μl/ウエル注入し、25℃で10分間維持した。その後、5N NaOH水溶液を100 μl/ウエルに注入し、反応を停止させた。
マイクロプレートリーダー(BIO−RAD社製、MICROPLATE READER Model 550(商品名))を用いて、405nm以上の吸光度を測定し、各試料に含まれるダニアレルゲンの濃度を定量した。試料を浸漬する前のダニアレルゲンの濃度(1μg/ml)と、試料を浸漬した後のダニアレルゲンの濃度から、以下の式に従い、試料のダニアレルゲン吸着率を算出した。
Claims (20)
- 前記金属フタロシアニンの誘導体が、金属フタロシアニンジカルボン酸、金属フタロシアニンテトラカルボン酸、金属フタロシアニンオクタカルボン酸、金属フタロシアニンジスルホン酸、金属フタロシアニンテトラスルホン酸、金属フタロシアニンオクタスルホン酸、またはこれら酸の塩であることを特徴とする請求項1に記載のアレルゲンの分解剤。
- 前記金属フタロシアニンの誘導体を、担体に担持または混合してあることを特徴とする請求項1に記載のアレルゲンの分解剤。
- 前記金属フタロシアニンの誘導体が、金属フタロシアニンジカルボン酸、金属フタロシアニンテトラカルボン酸、金属フタロシアニンオクタカルボン酸、金属フタロシアニンジスルホン酸、金属フタロシアニンテトラスルホン酸、金属フタロシアニンオクタスルホン酸、またはこれら酸の塩であることを特徴とする請求項5に記載のアレルゲンの分解方法。
- 前記ダニ由来のアレルゲンの分解剤は、前記金属フタロシアニンの誘導体を、担体に担持または混合してあることを特徴とする請求項5に記載のアレルゲンの分解方法。
- 前記塩が、ナトリウム塩または銅(II)塩であることを特徴とする請求項9に記載の抗アレルゲン羽毛。
- 前記金属フタロシアニンの誘導体の担持量が、前記羽毛の重量に対して、0.1質量%以上10質量%以下であることを特徴とする請求項9に記載の抗アレルゲン羽毛。
- 前記塩が、ナトリウム塩または銅(II)塩であることを特徴とする請求項13に記載の羽毛構造物。
- 前記金属フタロシアニンの誘導体の担持量が、前記羽毛重量に対して、0.1質量%以上10質量%以下であることを特徴とする請求項13に記載の羽毛構造物。
- 前記塩が、ナトリウム塩または銅(II)塩であることを特徴とする請求項17に記載の羽毛製品。
- 前記金属フタロシアニンの誘導体の担持量が、前記羽毛重量に対して、0.1質量%以上10質量%以下であることを特徴とする請求項17に記載の羽毛製品。
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JP2003382565 | 2003-11-12 | ||
JP2003382565 | 2003-11-12 | ||
JP2003387100 | 2003-11-17 | ||
JP2003387100 | 2003-11-17 | ||
PCT/JP2004/004445 WO2005047414A1 (ja) | 2003-11-12 | 2004-03-29 | アレルゲン分解剤、および抗アレルゲン羽毛 |
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JP2005337435A Division JP3885096B2 (ja) | 2003-11-12 | 2005-11-22 | アレルゲン分解繊維素材 |
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JP3858080B2 true JP3858080B2 (ja) | 2006-12-13 |
JPWO2005047414A1 JPWO2005047414A1 (ja) | 2007-08-23 |
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US (2) | US20080108593A1 (ja) |
EP (1) | EP1683847B1 (ja) |
JP (1) | JP3858080B2 (ja) |
KR (1) | KR101137731B1 (ja) |
CN (1) | CN1878845B (ja) |
AT (1) | ATE520756T1 (ja) |
WO (1) | WO2005047414A1 (ja) |
Cited By (1)
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KR101798676B1 (ko) | 2010-01-22 | 2017-11-16 | 세인트 조지스 하스피탈 메디컬 스쿨 | 먼지 진드기 그룹 1 펩티타제 알레르겐의 억제제로서 피루바미드 화합물 및 그의 용도 |
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JP4732123B2 (ja) * | 2005-10-28 | 2011-07-27 | ヤマセイ株式会社 | 加工羽毛の製造方法 |
KR20130086611A (ko) * | 2010-10-01 | 2013-08-02 | 가부시키가이샤 아이포레 | 화장료 조성물 |
CN103755713B (zh) * | 2014-01-27 | 2015-08-12 | 福州大学 | 一种八磺酸基酞菁及其制备方法和应用 |
US10617894B2 (en) * | 2016-04-05 | 2020-04-14 | Innonix Technologies, Incorporated | Compositions for reducing inhalation of toxic air pollution components |
BR102019020257A2 (pt) * | 2019-09-27 | 2021-04-20 | Mmf&T Desenvolvimento Tecnológico E Inovação Ltda | composição para saúde oral e processo de elaboração de composição para saúde oral |
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GB810548A (en) * | 1955-03-31 | 1959-03-18 | Hoechst Ag | Process for fixing pigments on fibrous materials or foils |
JPS5663355A (en) | 1979-10-31 | 1981-05-29 | Nippon Carbide Kogyo Kk | Deodorant |
JPS61258806A (ja) | 1985-05-11 | 1986-11-17 | Aasu Kuriin:Kk | 高分子物質を有効成分に含む消臭剤 |
JPS6468578A (en) * | 1987-09-02 | 1989-03-14 | Daiwa Spinning Co Ltd | Fiber having excellent washing fastness and deodorizing function |
JP2642106B2 (ja) * | 1987-10-20 | 1997-08-20 | 白井 汪芳 | 消臭繊維とその製造方法 |
JPH10117657A (ja) * | 1996-10-16 | 1998-05-12 | Masashi Fujii | ダニ類の誘引物質 |
JPH10273875A (ja) * | 1997-03-28 | 1998-10-13 | Kohjin Co Ltd | 抗菌防臭ならびに消臭性能を有する組成物 |
CN1197626C (zh) * | 1999-04-30 | 2005-04-20 | 吴羽化学工业株式会社 | 除臭结构体及除臭剂 |
JP4149111B2 (ja) * | 2000-02-02 | 2008-09-10 | 住化エンビロサイエンス株式会社 | 抗アレルゲン繊維及び繊維製品 |
US6465446B1 (en) * | 2001-03-12 | 2002-10-15 | John C. Dykstra | Treatment of dermatitis by the topical application of Δ5-androstene-3β-ol-7,17 dione and metabolizable precursors thereof |
JP2003213548A (ja) * | 2002-01-18 | 2003-07-30 | Atsushi Komiyama | 治療補助具 |
US6811796B2 (en) * | 2002-04-22 | 2004-11-02 | Matsuura Yakugyo Co., Ltd. | Preventive or therapeutic agent for pollen allergy, allergic rhinitis, atopic dermatitis, asthma or urticaria, or health food for prevention or improvement or reduction of symptoms thereof |
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- 2004-03-29 CN CN2004800332887A patent/CN1878845B/zh not_active Expired - Lifetime
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KR101798676B1 (ko) | 2010-01-22 | 2017-11-16 | 세인트 조지스 하스피탈 메디컬 스쿨 | 먼지 진드기 그룹 1 펩티타제 알레르겐의 억제제로서 피루바미드 화합물 및 그의 용도 |
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US20080108593A1 (en) | 2008-05-08 |
ATE520756T1 (de) | 2011-09-15 |
US20090291934A1 (en) | 2009-11-26 |
EP1683847B1 (en) | 2011-08-17 |
EP1683847A4 (en) | 2007-11-07 |
KR101137731B1 (ko) | 2013-11-28 |
CN1878845B (zh) | 2011-02-02 |
WO2005047414A1 (ja) | 2005-05-26 |
CN1878845A (zh) | 2006-12-13 |
EP1683847A1 (en) | 2006-07-26 |
JPWO2005047414A1 (ja) | 2007-08-23 |
KR20060112658A (ko) | 2006-11-01 |
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