JP2023519549A - 炎症性疾患の治療または改善のための竜眼肉含有混合生薬抽出物を含む経口用組成物およびその使用 - Google Patents
炎症性疾患の治療または改善のための竜眼肉含有混合生薬抽出物を含む経口用組成物およびその使用 Download PDFInfo
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Abstract
Description
乾燥竜眼肉(Longanae Arillus)(Buyoung Yakup Co.Ltd.)20g、乾燥藁本(Ligustici Tenuissimi Rhizoma)(Buyoung Yakup Co.Ltd.)20gおよび乾燥オンジ(Polygalae radix)(Buyoung Yakup Co.Ltd.)20gを小片に切断した後、6倍体積(v/w)の20%エタノール水溶液と混合し、その混合物を90±5℃で3日間還流抽出した。抽出物をろ紙(孔径10μm未満)を通して濾過させ、残渣を除去した後、残りの残渣を4倍体積(v/w)の20%エタノール水溶液でさらに2回抽出し、抽出物をろ紙(孔径、10μm未満)を通して濾過した。
実施例1に開示されたものとは異なる混合比および異なる溶媒を用いたことを除いては、全ての手順が実施例1と同様であり、竜眼肉(LA)、藁本(LT)およびオンジ(PR)の様々な本発明の混合抽出物、すなわち、本発明の混合抽出物(2)ないし本発明の混合抽出物(6)を得た。これを以下の実験にて試験試料として使用する。
本発明の抽出物の抗炎症活性を決定するために、文献(Jeong et al.,2019,J.Invest.Dermatol.,May;139 (5):pp1098-1109)に記載された手順に従って、HaCaT細胞を用いた以下のサイトカイン発現抑制試験を行った。
HT-29細胞およびTHP-1細胞に対する本発明の抽出物の細胞毒性を確認するために、HT-29細胞およびTHP-1細胞を用いた以下の細胞生存率は、当技術分野における既知の手順に従った。
HT-29細胞(ヒト結腸上皮細胞、韓国細胞株バンク、韓国細胞株研究財団、ソウル市鍾路区大学路101、03080、韓国)を10%小胎児血清と1%ペニシリン-ストレプトマイシン溶液を含有するDMEM培地に接種し、THP-1細胞(ヒト単核球細胞、韓国細胞株バンク、韓国細胞株研究財団、ソウル市鍾路区大学路101、03080、韓国)を10%小胎児血清および1%ペニシリン-ストレプトマイシン溶液を含有するRPMI培地に接種し、インキュベートした。
表4に示すように、HT-29細胞において本発明の抽出物(25~1000μg/mL)で処理した試験試料群の細胞生存率が培地のみで処理された陰性対照群と同様の細胞生存率を示しており、THP-1細胞では、500および1000μg/mL濃度でそれぞれ90.5%および26.5%を示し、陰性対照群と多少の差を示すことが確認され、500μg/mL未満の試験試料を以下の試験に適用した。
本発明の抽出物の抗炎症活性を決定するために、THP-1細胞を用いた以下のプロ炎症性サイトカインレベルの抑制試験を、当技術分野における既知の手順に従って行った。
3-1-1.試験手順
10、20、50、100、および500μg/mL脂質多糖類類(LPS)をヒト単核球細胞株(THP-1細胞韓国細胞株バンク、韓国細胞株研究財団、ソウル鍾路区大学路101、03080、韓国)に添加し、炎症モデルを作製した。
表5に示すように、実施例で調製された試験試料は、LPSを用量依存的に増加させながら増加したIL-1ベータの数値を大幅に減少させたことが確認できた。
3-2-1.試験手順
10、20、50、100、および500μg/mLの脂質多糖類(LPS)をヒト単核球細胞株(THP-1細胞韓国細胞株バンク、韓国細胞株研究財団、ソウル鍾路区大学路101、03080、韓国)に添加し、炎症モデルを作製した。
表6に示すように、実施例で調製された試験試料は、LPSを用量依存的に増加させながら増加したIL-10の数値を大幅に減少させることが確認できた。
本発明の抽出物が免疫細胞における炎症性因子の発現に及ぼす効果を測定するために、THP-1細胞を用いた以下の試験を、当技術分野における既知の手順に従って行った。
抗炎症活性とオートファジー経路の相関関係を確認するために、10、20、50、100、および500μg/mLの脂質多糖類(LPS)をヒト単核球細胞株(THP-1細胞韓国細胞株バンク、韓国細胞主研究財団、ソウル鍾路区大学路101、03080、韓国)に添加し、炎症モデルを作製した。
表7に示すように、試験試料と100μg/mLの脂質多糖類(LPS)で処理した試験群がLC3B-1とBeclin1の発現に対して用量依存的に増加する効果を示すことが確認された。
本発明の抽出物の関節炎抑制効果を確認するために、関節炎誘導性ラット動物モデルを用いた動物モデル実験を、当技術分野における既知の手順に従って行った。
MIA(ヨード酢酸モノナトリウム)誘導性変形性関節炎ラットモデルに対する試験試料の効能を評価するために、韓国、ソウルST.ソウル聖母病院カトリック大学に位置する「関節免疫疾患T2Bセンター(院長パク・ソンファン)」にて、以下の試験を行った。
5-1-1-1.動物モデル試験手順
(1)試験動物:12時間間隔で明暗サイクル(08:00~20:00)、温度21±2℃、相対湿度50±20%を維持しながら、ラット(オスウィスタラット、中央研究所動物、ソウル、韓国、7~8週齢、200~250g)をポリスルホン製ケージ(ケージ当たり2~3匹のマウス)で適切に管理された飼育室で飼育し、周囲の環境に順応させた。
(2)試験試料:実施例で調製した本発明の抽出物
(3)陽性対照群:セレコシブ(Celecoxib)(ハンリム製薬、ソウル、韓国)
(4)治療経路:変形性関節炎誘発後3日目に経口投与(1日1回)
(5)試験群の設定:表8参照。
(1)投与経路
MIAで変形性関節症を誘発した後、試験試料をビヒクル(食塩水)で処方された用量に応じて均質化し、1日1回1mLの試験試料を経口投与し、試験試料処理群(G2、G3)を調製した。
24匹のラットを6匹当たり4つの群に分け、試験試料と陽性対照群物質を所定の時間に経口投与した。
5-2-1.痛みの閾値の測定
5-2-1-1.痛みの閾値の測定方法
痛みの閾値(侵害受容潜伏期、閾値)は、痛みの閾値を測定するためのvon Freestyle評価方法に従い、一定期間、雄ウィスタラットの肢に加えられる力を徐々に増加させる装置である、ダイナミックプランター・エステシオメータ(Ugo Basile, 37400, Comerio, Italy)を用いて測定を行った(Kwon JY et al., Sci Rep. 2018 Sep 14;8(1):13832)。
痛みの測定試験の結果、100および150mg/kgの試験試料で処理した試験試料群は、用量依存的にビヒクルで処理した陰性対照群と比較して、強力な痛み抑制効果を示すことが確認された(表9-10)。
5-2-2-1.体重負荷の測定
正常な後肢(左)と関節炎誘発後肢(右)間の体重の変化(または体重分布)を測定するために、文献(Kwon JY et al.,Sci Rep.2018 Sep 14;8(1):13832)に記載の方法に従って試験装置(モデル600、IITC、USA)を用いて両後肢の負荷を測定した。
重量比(%)={右後肢の重量/(右後肢の重量+左後肢の重量)}×100
表11に示すように、試験試料群は、陰性対照群と比較して、両肢のバランス能力が大幅に向上されることが確認された。
本発明の抽出物の骨損傷抑制効果を決定するために、文献(Kwon JY et al.,Sci.Rep.2018 Sep 14;8(1):13832)に記載された公知の方法に従って組織学的解析を行った。
本発明の抽出物の変形性関節症による骨損傷に対する抑制効果を確認するために、犠牲にしたラット(雄ウィスタラット)の右膝関節にMIAと試験試料を投与し、ホルマリンで固定した。
表12に示すように、100および150mg/kgの試験試料で処理した試験試料群がマイクロCT撮影の試験結果を通じて陰性対象群(ビヒクル群)と比べて強力な骨損傷抑制効果を示すことが確認された。
本発明の抽出物の骨損傷抑制効果を決定するために、文献(Kwon JY et al.,Sci Rep.2018 Sep.14;8(1):13832)に公知された方法に従って以下の組織病理学的解析を行った。
本発明の抽出物の変形性関節症による骨損傷に対する抑制効果を確認するために、犠牲にしたラット(雄ウィスタラット)の右膝関節にMIAと試験試料を投与し、ホルマリンで固定されたラットの右膝関節をスライスし、Safranin Oで染色した。
表13(総マンキンスコア法)および表14(OARSIスコア)からわかるように、100および150mg/kgの試験試料で処理した試験試料群は、組織病理学的解析結果を通じた陰性対照群群(ビヒクル群)に比べて骨損傷に対する強力な抑制効果を示すことが確認された。
すべてのデータは、平均および標準偏差(平均±SD)で表され、GraphPad PRISMバージョン5.0(USA)解析プログラムにより、二元配置分散分析(two-way ANOVA)、一元配置分散分析(one-way ANOVA)を用いて統計的有意性検証が有意であると判断(P<0.05)された。
WIN-1001Xエキス:100mg
メタ重亜硫酸ナトリウム:3.0mg
メチルパラベン:0.8mg
プロピルパラベン:0.1mg
注入用蒸留水:最適量
有効成分を溶解させ、pHを約7.5に調整した後、全成分を2mlアンプルに充填し、従来の注射剤調製法により滅菌して注射剤を調製した。
WIN-1002Xエキス:500mg
トウモロコシデンプン:100mg
ラクトース:100mg
タルク:10mg
上記の成分を混合し、密封パッケージに充填することにより散剤の製剤を調製した。
WIN-1003Xエキス:200mg
トウモロコシデンプン:100mg
ラクトース:100mg
ステアリン酸マグネシウム:最適量
前記の成分を混合し、錠剤化することにより錠剤の製剤を調製した。
WIN-1004Xエキス:100mg
ラクトース:50mg
トウモロコシデンプン:50mg
タルク:2mg
ステアリン酸マグネシウム:最適量
上記の成分を混合し、ゼラチンカプセルを従来のゼラチン製造方法により充填して錠剤の製剤を調製した。
WIN-1005Xエキス:1000mg
糖:20g
多糖類:20g
レモン風味:20g
有効成分を溶解させた後、全成分を1000mlのアンプルに充填し、従来の液体の調製法で滅菌し、調製液を調製した。
WIN-1001Xエキス:1000mg
ビタミン混合物:最適量
ビタミンAアセテート:70g
ビタミンE:1.0mg
ビタミンB10:13mg
ビタミンB2:0.15mg
ビタミンB6:0.5mg
ビタミンB1:20.2g
ビタミンC:10mg
ビオチン:10g
アミドニコチン酸:1.7mg
葉酸:50g
パントテン酸カルシウム:0.5mg
ミネラル混合物:最適量
硫酸第一鉄:1.75mg
酸化亜鉛:0.82mg
炭酸マグネシウム:25.3mg
リン酸一カリウム:15mg
リン酸二カルシウム:55mg
クエン酸カリウム:90mg
炭酸カルシウム:100mg
塩化マグネシウム:24.8mg
上記のビタミンとミネラルの混合物は、あらゆる方法で多様にできる。このような変形は、本発明の精神および範囲から逸脱するとみなされるべきではない。
WIN-1002Xエキス:1000mg
クエン酸:1000mg
オリゴ糖:100g
アプリコット濃度:2g
タウリン:1g
蒸留水:900ml
有効成分を溶解させて混合し、85℃で1時間撹拌した後、濾過した後、全成分を1000mlアンプルに充填し、既存の健康飲料の調製方法で滅菌し、健康飲料の製剤を調製した。
Claims (8)
- 皮膚炎、アトピー性皮膚炎、結膜炎、歯周炎、鼻炎、中耳炎、咽頭痛、扁桃炎、肺炎、胃潰瘍、胃炎、クローン病、大腸炎、痔、痛風、リウマチ熱、狼瘡、線維筋痛症、腱炎、腱鞘炎、腱周囲炎、筋炎、肝炎、膀胱炎、腎炎、シェーグレン症候群、慢性炎症および急性炎症によって引き起こされる掻痒症の群から選択される炎症性疾患を予防および治療するための竜眼肉(Longanae Arillus)、藁本(Ligustici Tenuissimi Rhizoma)およびオンジ(Polygalae radix)の混合生薬抽出物を有効成分として含む経口用医薬組成物。
- 前記混合生薬抽出物は、(a)0.01-100:0.01-100:0.01-100重量部(w/w)の範囲の竜眼肉、藁本およびオンジの乾燥重量(w/w)を基準とした混合比を有する竜眼肉、藁本およびオンジの混合生薬抽出物、または(b)0.01-100:0.01-100:0.01-100重量部(w/w)の範囲の竜眼肉、藁本およびオンジの乾燥重量(w/w)を基準とした混合比を有する竜眼肉、藁本およびオンジの各抽出物の混合物であることを特徴とする、請求項1に記載の経口用医薬組成物。
- 前記抽出物は、水、C1-C4低級アルキルアルコール、例えば、メタノール、エタノール、プロパノール、ブタノール、アセトン、酢酸エチル、クロロホルム、ヘキサン、ブチレングリコール、プロピレングリコールまたはグリセリンから選択される1つ以上の溶媒で抽出されることを特徴とする、請求項1に記載の経口用医薬組成物。
- 皮膚炎、アトピー性皮膚炎、結膜炎、歯周炎、鼻炎、中耳炎、咽頭痛、扁桃炎、肺炎、胃潰瘍、胃炎、クローン病、大腸炎、痔、痛風、リウマチ熱、狼瘡、線維筋痛症、腱炎、腱鞘炎、腱周囲炎、筋炎、肝炎、膀胱炎、腎炎、シェーグレン症候群、慢性炎症および急性炎症によって引き起こされる掻痒症の群から選択される炎症性疾患を予防および改善するための竜眼肉(Longanae Arillus)、藁本(Ligustici Tenuissimi Rhizoma)およびオンジ(Polygalae radix)の混合生薬抽出物を有効成分として含む健康機能食品。
- 前記健康機能食品は、散剤、顆粒剤、錠剤、カプセル剤、丸剤、懸濁剤、乳剤、シロップ、ティーバッグ、浸出茶、または飲料の形態で提供されることを特徴とする、請求項4に記載の健康機能食品。
- 皮膚炎、アトピー性皮膚炎、結膜炎、歯周炎、鼻炎、中耳炎、咽頭痛、扁桃炎、肺炎、胃潰瘍、胃炎、クローン病、大腸炎、痔、痛風、リウマチ熱、狼瘡、線維筋痛症、腱炎、腱鞘炎、腱周囲炎、筋炎、肝炎、膀胱炎、腎炎、シェーグレン症候群、慢性炎症および急性炎症によって引き起こされる掻痒症の群から選択される炎症性疾患を予防または改善するための竜眼肉(Longanae Arillus)、藁本(Ligustici Tenuissimi Rhizoma)およびオンジ(Polygalae radix)の混合生薬抽出物を有効成分としてまたは主要成分として含む健康補助食品。
- 竜眼肉(Longanae Arillus)、藁本(Ligustici Tenuissimi Rhizoma)およびオンジ(Polygalae radix)の有効量の混合生薬抽出物またはその薬学的に許容される担体を哺乳動物に経口投与する段階を含む、哺乳動物の皮膚炎、アトピー性皮膚炎、結膜炎、歯周炎、鼻炎、中耳炎、咽頭痛、扁桃炎、肺炎、胃潰瘍、胃炎、クローン病、大腸炎、痔、痛風、リウマチ熱、狼瘡、線維筋痛症、腱炎、腱鞘炎、腱周囲炎、筋炎、肝炎、膀胱炎、腎炎、シェーグレン症候群、慢性炎症および急性炎症によって引き起こされる掻痒症の群から選択される炎症性疾患を治療または改善する方法。
- 有効成分であって、皮膚炎、アトピー性皮膚炎、結膜炎、歯周炎、鼻炎、中耳炎、咽頭痛、扁桃炎、肺炎、胃潰瘍、胃炎、クローン病、大腸炎、痔、痛風、リウマチ熱、狼瘡、線維筋痛症、腱炎、腱鞘炎、腱周囲炎、筋炎、肝炎、膀胱炎、腎炎、シェーグレン症候群、慢性炎症および急性炎症によって引き起こされる掻痒症の群から選択される炎症性疾患を治療または改善するために使用される経口用製剤を調製するための 竜眼肉(Longanae Arillus)、藁本(Ligustici Tenuissimi Rhizoma)およびオンジ(Polygalae radix)の混合生薬抽出物の使用。
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KR1020210032875A KR102374384B1 (ko) | 2020-04-03 | 2021-03-12 | 용안육 포함 조합 생약 추출물을 유효성분으로 함유하는 염증질환의 치료 또는 예방용 조성물 |
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PCT/KR2021/003728 WO2021201502A1 (en) | 2020-04-03 | 2021-03-25 | An oral pharmaceutical composition comprising an extract of combined herbs comprising longanae arillus for the treatment or alleviation of inflammatory disease and the use thereof. |
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JP4105498B2 (ja) * | 2002-08-02 | 2008-06-25 | 株式会社ノエビア | アトピー性疾患の症状の予防・緩和に有効な組成物 |
KR100728069B1 (ko) * | 2004-07-09 | 2007-06-13 | 서미자 | 한방 기능성 음료의 제조방법 및 그 음료 |
KR100742378B1 (ko) * | 2005-05-17 | 2007-07-24 | 김정진 | 생약재를 함유하는 아토피성 피부염 치료용 조성물 |
KR101051362B1 (ko) * | 2007-08-31 | 2011-07-22 | (주)콤비메드 | 생약재 발효산물을 포함하는 아토피성 피부염 치료 및/또는예방용 조성물 |
KR101047644B1 (ko) * | 2008-08-06 | 2011-07-07 | (주)아모레퍼시픽 | 포공영, 승마, 산약 또는 고본 추출물을 함유하는 피부외용제 조성물 |
CN101670080A (zh) * | 2008-09-11 | 2010-03-17 | 北京天科仁祥医药科技有限公司 | 一种用于慢性胃炎中药组合及其制备工艺 |
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CN103083376B (zh) * | 2012-06-01 | 2015-08-19 | 吉林大学 | 可用于治疗鼻炎的藁本提取物 |
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CN103520681B (zh) * | 2013-10-18 | 2016-01-20 | 河南中医学院 | 一种治疗脾肾阳虚型慢性结肠炎的中药 |
KR20150086982A (ko) * | 2014-01-21 | 2015-07-29 | 경희대학교 산학협력단 | 원지 추출물을 유효성분으로 함유하는 아토피 피부염의 예방 또는 치료용 조성물 |
CN104147378B (zh) * | 2014-07-21 | 2018-06-12 | 广西梧州三鹤药业有限公司 | 治疗血虚风燥型牛皮癣的中药组合物 |
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CN104887859A (zh) * | 2015-06-18 | 2015-09-09 | 王峰 | 一种治疗结膜炎的中药剂 |
CN106237208A (zh) * | 2016-08-24 | 2016-12-21 | 韦克康 | 治疗慢性鼻炎的中药丸及其制备方法 |
KR101867530B1 (ko) * | 2016-12-30 | 2018-06-14 | 한국과학기술연구원 | 용안육 추출물을 함유하는 숙취 예방 또는 해소용 조성물 |
KR102126470B1 (ko) * | 2018-05-28 | 2020-06-24 | 주식회사 엘지생활건강 | 용안육 추출물을 포함하는 화장료 조성물 |
CN108936675A (zh) * | 2018-08-16 | 2018-12-07 | 刘守峰 | 一种具有治疗痛风作用的保健食品及其制备方法 |
CN109453157B (zh) * | 2018-10-21 | 2020-10-09 | 魏群 | 一种重症急性胰腺炎肠黏膜屏障保护药物及其用途 |
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