JP2016519101A - オメガ−3脂肪酸およびトマトリコペンを含む抗炎症性相乗的組み合わせ - Google Patents
オメガ−3脂肪酸およびトマトリコペンを含む抗炎症性相乗的組み合わせ Download PDFInfo
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Abstract
Description
本発明は、特に、オメガ−3脂肪酸とリコペンとの相乗的組み合わせを含む組成物を対象とする。より具体的には、本発明は、特に炎症を阻害/抑制するために用いられ得る前述の化合物の相乗的組み合わせを含む組成物を提供する。
非特異的免疫系の重要な部分を形成する炎症プロセスは、微生物および他の潜在的に有害な環境要因に直面した宿主防御にとって不可欠な化学変化および細胞変化の複雑な組み合わせを特徴とする。しかし多くの例で、炎症は、不適切に惹起されること、および/または宿主にとって有害になる程度に持続することがある。そのような例では、特に非感染性炎症疾患の場合に、炎症プロセスの1つ以上の態様の発生を阻害または予防することが必要になり得る。
一実施形態において、本発明は、トマトリコペンまたはLyc−O−Mato(登録商標)、オメガ−3脂肪酸、およびカルノシン酸を含む組成物を提供する。別の実施形態において、本発明は、トマトリコペンまたはLyc−O−Mato(登録商標)、オメガ−3脂肪酸、ルテインおよびカルノシン酸を含む組成物をさらに提供する。別の実施形態において、本発明は、該組成物がルテイン、フィトエン、フィトフルエン、β−カロテン、トコフェロール、フィトステロール、またはそれらの任意の組み合わせをさらに含むことをさらに提供する。別の実施形態において、本発明は、オメガ−3脂肪酸とトマトリコペンとのモル濃度比が2000:1〜10:1であることをさらに提供する。別の実施形態において、本発明は、オメガ−3脂肪酸とカルノシン酸とのモル濃度比が1500:1〜2:1であることをさらに提供する。別の実施形態において、本発明は、オメガ−3脂肪酸とトマトリコペンとカルノシン酸とのモル濃度比が2000:1:5〜2:1:1であることをさらに提供する。別の実施形態において、本発明は、オメガ−3脂肪酸がROPUFA(登録商標)であることをさらに提供する。別の実施形態において、本発明は、オメガ−3脂肪酸が、ドコサヘキサエン酸、エイコサペンタエン酸またはそれらの組み合わせであることをさらに提供する。別の実施形態において、本発明は、該組成物が相乗的抗炎症効果を有することをさらに提供する。
一実施形態において、本発明は、オメガ−3脂肪酸およびトマトリコペンを含む組成物を提供する。一実施形態において、本発明は、オメガ−3脂肪酸とトマトリコペンとの相乗的組み合わせ(以下にさらに説明される)を含む組成物を提供する。一実施形態において、本発明は、オメガ−3脂肪酸、カルノシン酸、およびトマトリコペンを含む組成物を提供する。一実施形態において、本発明は、オメガ−3脂肪酸とカルノシン酸とトマトリコペンとの相乗的組み合わせ(以下にさらに説明される)を含む組成物を提供する。一実施形態において、本発明は、オメガ−3脂肪酸、ルテイン、およびトマトリコペンを含む組成物を提供する。一実施形態において、本発明は、オメガ−3脂肪酸、ルテイン、カルノシン酸、およびトマトリコペンを含む組成物を提供する。一実施形態において、本発明は、オメガ−3脂肪酸とルテインとカルノシン酸とトマトリコペンとの相乗的組み合わせ(以下にさらに説明される)を含む組成物を提供する。
一般に、本明細書で用いられる命名法および本発明で用いられる実験手順は、化学、分子学、生化学、および細胞生物学の技術を含む。そのような技術は、文献に完全に説明されている。例えば、”Molecular Cloning: A laboratory Manual” Sambrook et al., (1989); ”Current Protocols in Molecular Biology” Volumes I−III Ausubel, R. M., ed. (1994); ”Cell Biology: A Laboratory Handbook”, Volumes I−III Cellis, J. E., ed. (1994); The Organic Chemistry of Biological Pathways by John McMurry and Tadhg Begley (Roberts and Company, 2005); Organic Chemistry of Enzyme−Catalyzed Reactions by Richard Silverman (Academic Press, 2002); Organic Chemistry (6th Edition) by Leroy ”Skip” G Wade; Organic Chemistry by T. W. Graham Solomons and, Craig Fryhleを参照されたい。
細胞
マクロファージ単離物および培養腹腔マクロファージを、採取4日前にチオグリコラートブロス(4%)1.5mlの腹腔注射後の6〜8週齢の雄ICRマウス(Harlan,イスラエル所在)の腹腔から採取した。腹腔マクロファージは、リン酸緩衝生理食塩水(PBS)で3回洗浄し、適宜、赤血球の低張溶解を実施して、高濃度(90〜95%)マクロファージ細胞集団を得た。マクロファージを、フローマイクロフルオリメトリー(flow microfluorimetry)によるFITCコンジュゲート化抗マウスF4/80(MCA497F; Serotec,英国オックスフォード所在)を用いたFACS(Becton−Dickinson,米国カリフォルニア州マウンテンビュー所在)分析により同定した。各試料で、10,000個の光散乱でゲートされた生存細胞を分析した。腹腔マクロファージ(1×106細胞/ウェル)を10%ウシ胎仔血清、2mMのL−グルタミン、100U/mlペニシリン、100mg/mlストレプトマイシン(Beit−Haemek,イスラエル所在)を含むRPMI1640培地中、5%CO2雰囲気および37℃で96ウェルプレートにて培養した。細胞を、本明細書に記載された異なる組み合わせの非存在下または存在下で、ネズミチフス菌由来の1mg/mlのLPSで刺激した(実施例の節の図を参照)。植物栄養素をジメチルスルホキシド(DMSO;最終濃度5mM)に溶解した。混合物をボルテックスにかけて、37℃の水浴で10分間インキュベートし(振とうしながら)、ソニケータバス中で15秒ずつ3回音波処理した。該化合物の希釈標準濃度を、原液から、加温した培地に適切な容量を添加することにより調製した。溶液の最終濃度を、イソプロパノール0.5mlと、0.025%BHTを含むヘキサン/ジクロロメタン(1/5v/v)1.5mlを培地1mlに添加することにより計算した。溶液をボルテックスにかけて、液相を3000rpmでの10分間の遠心分離により分離した。分光光度法を実施して、リコペン、アスタキサンチン、ルテイン、β−カロテンの濃度を測定した。対照に適切な容量のDMSO(0.1〜0.2%)を添加し、各試験管における阻害率を、その対照を基準として計算した。
1.スーパーオキシドアニオン(O2 −)の放出を、マイクロタイタープレート技術によりフェリシトクロムcのスーパーオキシドジスムターゼで阻害可能な還元として測定した。フェリシトクロムcの還元は、Thermomaxマイクロプレートリーダー(Molecular Devices,米国カリフォルニア州メンローパーク所在)により550nmでの吸光度変化を2分間隔で30分間測定することにより追跡した。スーパーオキシド産生の最大速度を決定して、吸光係数E550=21mM/cmを用いてnmol O2 −/106細胞/10分間として表した。
2.スーパーオキシドの内部産生を、ジヒドロローダミン123(DHR123)の還元により検出した:ハンクス液(pH7.4)490ml中の細胞3×105個を、ポリプロピレン試験管中で最終濃度1mMのDHR123 5mlと共に37℃で5分間インキュベートした。刺激された細胞および非刺激の細胞を、FACS(BeckmanCoulter,米国カリフォルニア州フラートン所在)により直ちに分析した。
オメガ−3によるNO産生の用量依存的阻害
本明細書に記載された細胞への抗炎症効果を、LPSに刺激されたマクロファージによるNO産生の阻害により検出した。
オメガ−3およびトマトリコペンによるNO産生の用量依存性相乗的阻害
本明細書に記載された細胞への抗炎症効果を、LPSに刺激されたマクロファージによるNO産生の阻害により検出した。
オメガ−3、カルノシン酸、およびトマトリコペンによるNO産生の用量依存性相乗的阻害
本明細書に記載された細胞への抗炎症効果を、LPSに刺激されたマクロファージによるNO産生の阻害により検出した。
オメガ−3、カルノシン酸、およびルテインによるNO産生の用量依存性相乗的阻害
本明細書に記載された細胞への抗炎症効果を、LPSに刺激されたマクロファージによるNO産生の阻害により検出した。
オメガ−3、カルノシン酸、ルテイン、およびトマトリコペンによるNO産生の用量依存性相乗的阻害
本明細書に記載された細胞への抗炎症効果を、LPSに刺激されたマクロファージによるNO産生の阻害により検出した。
さらなるオメガ−3単独による、またはトマトリコペンとの組み合わせによるNO産生の用量依存性相乗的阻害
本明細書に記載された細胞への抗炎症効果を、LPSに刺激されたマクロファージによるNO産生の阻害により検出した。
Claims (17)
- オメガ−3脂肪酸、カルノシン酸、およびトマトリコペンを含む組成物。
- 前記組成物が、フィトエン、フィトフルエン、β−カロテン、トコフェロール、フィトステロール、またはそれらの任意の組み合わせをさらに含む、請求項1に記載の組成物。
- オメガ−3脂肪酸とトマトリコペンとのモル濃度比が2000:1〜10:1である、請求項1に記載の組成物。
- オメガ−3脂肪酸とカルノシン酸とのモル濃度比が1500:1〜2:1である、請求項1に記載の組成物。
- オメガ−3脂肪酸とトマトリコペンとカルノシン酸とのモル濃度比が2000:1:5〜2:1:1である、請求項1に記載の組成物。
- 前記組成物が相乗的抗炎症効果を有する、請求項1に記載の組成物。
- ルテインをさらに含む、請求項1に記載の組成物。
- 前記組成物が経口組成物である、請求項1に記載の組成物。
- 医薬的に許容し得る賦形剤をさらに含む、請求項1に記載の組成物。
- 前記オメガ−3脂肪酸が、ドコサヘキサエン酸、エイコサペンタエン酸、またはそれらの組み合わせである、請求項1に記載の組成物。
- 炎症に見舞われた対象を処置する方法であって、オメガ−3脂肪酸と、トマトリコペンとを含む組成物の治療有効量を前記対象に投与し、それにより炎症に見舞われた対象を処置する工程を含む、前記方法。
- オメガ−3脂肪酸とトマトリコペンとのモル濃度比が2000:1〜10:1である、請求項11に記載の方法。
- 前記オメガ−3脂肪酸が、ドコサヘキサエン酸、エイコサペンタエン酸、またはそれらの組み合わせである、請求項11に記載の方法。
- 前記組成物が、経口組成物である、請求項11に記載の方法。
- 炎症に見舞われた対象を処置する方法であって、請求項1〜10のいずれか1項に記載の組成物の治療有効量を前記対象に投与し、それにより炎症に見舞われた対象を処置する工程を含む、前記方法。
- 前記炎症に見舞われた対象を処置することが、前記対象におけるNO、PGE、TNF−α、またはそれらの任意の組み合わせの産生を阻害することである、請求項11〜15のいずれか1項に記載の方法。
- 前記炎症に見舞われた対象を処置することが、前記対象における炎症部位での好中球の動員を阻害すること、炎症部位での好中球の活性化を阻害すること、またはそれらの組み合わせである、請求項11〜15のいずれか1項に記載の方法。
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