JP5504534B2 - ケトン体生成促進剤組成物 - Google Patents
ケトン体生成促進剤組成物 Download PDFInfo
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- JP5504534B2 JP5504534B2 JP2009507548A JP2009507548A JP5504534B2 JP 5504534 B2 JP5504534 B2 JP 5504534B2 JP 2009507548 A JP2009507548 A JP 2009507548A JP 2009507548 A JP2009507548 A JP 2009507548A JP 5504534 B2 JP5504534 B2 JP 5504534B2
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- hydroxybutyric acid
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Description
以上のように、1回の投与で短時間に血中濃度が大きく数倍以上に上昇し、数時間は維持されるとしている。
本発明の組成物は特に脳細胞損傷に起因する疾患の治療および/または予防、特に脳卒中の予防または治療において有用である。また本発明の組成物は脳卒中の後遺症からの回復の促進に有用である。
健康食品として市販されているスピルリナは通常、その乾燥重量中0.1重量%程度以下のβ−ヒドロキシ酪酸重合体しか含有しない。本発明においては培養条件を調節する等によりβ−ヒドロキシ酪酸重合体含有量を増したスピルリナ(例えばJ.Biotechnol.,172,2791-2792(1990)参照、本文献は引用により本願に含まれる)が好適に用いられる。本発明の組成物においてβ−ヒドロキシ短〜中鎖脂肪酸重合体としては、スピルリナから単離したものを用いても、スピルリナをそのまま配合してもよい。
本発明に用いられるβ−ヒドロキシ短〜中鎖脂肪酸重合体は、真核生物の細胞膜やミトコンドリアに存在することが知られている。動物では血液中にアルブミン等と結合して存在することも既知である。(Eur.J.Biochem.,224,317-328(1994))
本発明の組成物は特に脳細胞損傷に起因する疾患の治療および/または予防、特に脳卒中の予防または治療において有用である。また、本発明の組成物は、脳卒中の後遺症、例えば麻痺、痺れ、痛み、言語障害、視覚障害、感覚障害、尿失禁および情緒障害など、特に麻痺、痺れ、痛み、尿失禁からの回復促進に好適に用いられる。
また、飼料へ配合して動物に投与させるための飼料添加剤として調製してもよい。
さらに、ヌクレオチドなどのリン酸化合物とのリン酸エステルも好適に用いられる。
実施例1:分子量と水溶性・加水分解性
発酵により製造した(β−ヒドロキシ酪酸−β−ヒドロキシ吉草酸)共重合体(GPCポリスチレン換算重量平均分子量Mw=560,000。数平均分子量Mn=238,000。β−ヒドロキシ吉草酸=5.4モル%<NMR測定>。PHBV−1)10gをクロロホルム250mlに溶解させ、濃塩酸2mlを加えて7時間加熱還流した。次いで30%過酸化水素水を6.4ml添加し、加熱環流を更に8時間続けた。水で中性になるまで洗浄した後、クロロホルム層を回収しエバポレーションしてPHBV−2を回収した。ゲルパーミエーションクロマトグラフィー(東ソー(株)GPC HLC-8020/カラムTSKgel MultiporeHXL-M )で重量平均分子量=13,600、数平均分子量=5,600(ポリスチレン換算)と測定された。
一方、PHBV−4は水中へ投入した2時間後には、目視でほぼ全量が溶解したことが確認され、PHBV−4が水溶性であることが確認された。
ペプシン:胃液類似液(塩酸水溶液pH=1.5)
アミラーゼ:胃液類似液(塩酸水溶液pH=5.5、塩化カルシウム0.03重量%)
トリプシン:腸液類似液(炭酸水素ナトリウム水溶液pH=8.0)
リパーゼ:腸液類似液(炭酸水素ナトリウム水溶液pH=7.5、塩化カルシウム0.03重量%)
使用動物:Sprague-Dawley系雄性SPFラット(日本エスエルシー(株)、浜松)8週齢(n=3)
基礎飼料:市販粉末飼料CE−2(日本クレア(株),東京)
給水:自由摂取
飼育環境:温度23℃、湿度50%、毎日8時〜20時に蛍光灯点灯。
飼育:4日間施設馴致後群分けし、対照群・試験群ともに、基礎飼料を10時から17時まで自由摂取させる時間制限給餌に7日間馴致した。試験開始日の給餌直前に尾部より末梢血を採血した。対照群には引続き基礎飼料を、試験群には基礎飼料に(β−ヒドロキシ酪酸−β−ヒドロキシ吉草酸)共重合体(GPCポリスチレン換算重量平均分子量=560,000。β−ヒドロキシ吉草酸=5.4モル%。PHBV−1)を5wt%添加したものを、時間制限給餌した。4時間毎に尾部より末梢血の採血を行い、血中のβ−ヒドロキシ酪酸の濃度変化を観察した。血中β−ヒドロキシ酪酸濃度は常法により得られた末梢血から血清を分離し、これをPBS(pH7.4)で希釈したものを(株)日本医学臨床検査研究所(京都)にて酵素サイクリング法で測定した。
使用動物:Sprague-Dawley系雄性ラット(SPF)(日本エスエルシー(株)、浜松)8週齢(n=3)
基礎飼料:市販粉末飼料CE−2(日本クレア(株),東京)
給水:自由摂取
飼育環境:温度23℃、湿度50%、毎日8時〜20時に蛍光灯点灯。
飼育:4日間施設馴致後群分けし、対照群は基礎飼料を、試験群は基礎飼料にPHBV−1を5重量%添加したものを2週間自由摂取させた。最終日の朝解剖し、腹部大動脈から採血し血清を得た。肝臓は、氷冷下細断してよく混合し、0.1gをマイクロチューブに投入した。PBS(pH7.4)に0.1mMのEDTA2Naを加えた溶液を0.9ml加えて、氷冷下、肝臓組織をホモジナイズした。遠心分離後後、上澄を得た。血清と上澄中のケトン体(アセト酢酸AcAcおよびβ−ヒドロキシ酪酸HB)を(株)日本医学臨床検査研究所(京都)において酵素サイクリング法で測定した。
使用動物:脳卒中易発性高血圧ラットSHRSP/Izm(SPF)(日本エスエルシー(株)、浜松) 雄性9週齢(n=5)
基礎飼料:市販粉末飼料SP−2(日本エスエルシー(株)、浜松)に1wt%の食塩を添加
給水:自由摂取
飼育環境:温度23℃、湿度50%、毎日8時〜20時に蛍光灯点灯。
飼育:4日間施設馴致後群分けし、対照群は基礎飼料を、試験群は基礎飼料にPHBV−1を5重量%添加したものを自由摂取させた。前肢を持上げる、横転するなどの脳卒中の発症が認められるまで飼育し、その発症までの期間を調べた。
対照群の発症までの期間=15,16,27,35,36日
試験群の発症までの期間=23,26,31,37,50日
試験群の方が、常に発症が遅くPHBV−1の脳卒中予防における効果が示唆された。
患者1
出血性脳卒中発症後、4年1ヶ月経過した50歳男性。右半身に麻痺と痺れや疼痛の後遺症があり2級身体障害者認定を受けている。
β−ヒドロキシ短〜中鎖脂肪酸重合体の投与
容量0.3gの1号カプセルに充填したβ−ヒドロキシ酪酸重合体(発酵で製造。重量平均分子量839,000)を1日に1カプセル服用させた。
結果
投与開始4日目から痺れが軽減する時間が現れ始めた。2週間後には痺れや鈍痛が軽くなったことを明白に自覚するようになった。1日のうちで痺れ感や鈍痛のない時間が徐々に増え、6ヶ月後には全く解消した。
この痺れや鈍痛の軽減に並行して、筋肉や身体の緊張が弛むのを感じるようになった。この弛緩(リラクセーション)によって、麻痺している部位を自分で動かしたという主動感を持って動かすことができるようになった。
リハビリテーションを重ねることで動きが徐々によくなり、3ヶ月後には、右腕や右足などの麻痺が軽減したのが実感でき、自力でほぼ通常生活を送ることができるようになった。
さらに痺れと同時に尿失禁があったが、β−ヒドロキシ酪酸重合体服用開始後8日目から失禁回数が急速に減り、2週間後には全く尿失禁が無くなった。
患者2
出血性脳卒中発症後、3年経過した60歳男性。右足に麻痺及び鈍痛、唇の右側に麻痺の後遺症あり。
β−ヒドロキシ短〜中鎖脂肪酸重合体の投与
容量0.3gの1号カプセルに充填した、上記β−ヒドロキシ酪酸重合体を1日に1カプセル服用させた。
結果
投与開始8日後には痺れと鈍痛が軽減するのを自覚した。10日間の服用で止めたが、症状がもとに戻ることはなかった。
スピルリナ(Spirulina platensis)を、18リットルのKosaric培地に0.5W/V%の酢酸ナトリウムを加えた培地に入れ(スピルリナ乾燥重量として0.125g/L,プラスチック製水槽使用)、光照射下(1日16時間、水面で900〜1100ルクス)水温24℃でpHを9.3〜9.6に保つように空気をバブリングしながら25日間培養した。濾過にて採集してオーブン(70℃)で乾燥して43gの乾燥スピルリナを得た。この乾燥スピルリナを用いてクロロホルム抽出法で、β−ヒドロキシ酪酸重合体の含有量を調べたところ3.1重量%であった。抽出物はNMRを用いてβ−ヒドロキシ酪酸重合体であることを確認した。
微量ミネラル水溶液(成分(g/L)):H2BO4(2.86),MnCl2・4H2O(1.81),ZnSO4・7H2O(0.22),CuSO4・5H2O(0.08),MoO3(0.01)
Claims (10)
- β−ヒドロキシ酪酸の単独重合体およびβ−ヒドロキシ酪酸と炭素数3〜12のβ−ヒドロキシ飽和脂肪酸との共重合体からなる群から選択される重合度10以上の水不溶性重合体を含む、ヒトを含む動物においてケトン体生成促進するための経口投与用、経鼻投与用、または消化管内投与用医薬組成物。
- 炭素数3〜12のβ−ヒドロキシ飽和脂肪酸が、β−ヒドロキシプロピオン酸、β−ヒドロキシ吉草酸、β−ヒドロキシカプロン酸、β−ヒドロキシカプリル酸、β−ヒドロキシカプリン酸およびこれらの混合物からなる群から選択される、請求項1記載の組成物。
- β−ヒドロキシ酪酸の単独重合体を含む、請求項1または2に記載の組成物。
- β−ヒドロキシ酪酸とβ−ヒドロキシ吉草酸の共重合体を含む、請求項1〜3いずれかに記載の組成物。
- β−ヒドロキシ酪酸の単独重合体およびβ−ヒドロキシ酪酸と炭素数3〜12のβ−ヒドロキシ飽和脂肪酸との共重合体からなる群から選択される重合度10以上の水不溶性重合体の重量平均分子量が1,000〜20,000,000である、請求項1〜4いずれかに記載の組成物。
- β−ヒドロキシ酪酸の単独重合体およびβ−ヒドロキシ酪酸と炭素数3〜12のβ−ヒドロキシ飽和脂肪酸との共重合体からなる群から選択される重合度10以上の水不溶性重合体がスピルリナにより産生されたものである、請求項1〜5いずれかに記載の組成物。
- アルツハイマー病、ピック病関連前頭側頭骨変性、血管性痴呆、レーヴィ体型の老人性痴呆、前頭萎縮を伴う振せん麻痺の痴呆、進行性核上性麻痺および大脳皮質基底核変性症、ダウン症候群関連アルツハイマー、重症筋無力症ならびに筋ジストロフィーからなる群から選択される神経変性疾患、筋萎縮性側索硬化症、てんかん、パーキンソン病、フリーラジカル疾患、心不全、心筋梗塞、狭心症、II型糖尿病、ピルビン酸デヒドロゲナーゼの欠乏もしくは遮断、1又はそれ以上の種類の細胞型における解糖の実施不能及びデュシェンヌ型筋ジストロフィー、細胞代謝にとって有害な遺伝的欠陥、インシュリン耐性又は他のグルコース代謝欠陥もしくは欠陥誘発性状態、GABA予防発作からなる群から選択される疾患乃至症状の治療または予防;虚血または頭部外傷が介在する脳疾患状態の抑制または治療;または脳浮腫の抑制、脳機能の保護、脳エネルギー代謝の調整、および脳卒中の程度の軽減から選択される脳機能改善のために用いられる、請求項1〜6いずれかに記載の組成物。
- 脳細胞損傷に起因する疾患の治療および/または予防のために用いられる、請求項1〜6いずれかに記載の組成物。
- 脳卒中の予防または治療のために用いられる、請求項8記載の組成物
- 脳卒中の後遺症からの回復促進のために用いられる、請求項8記載の組成物。
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CA3223325A1 (en) * | 2021-06-28 | 2023-01-05 | Roberto Vianna Nonato | Methods to produce therapeutic formulations comprising hydroxybutirate and hydroxyvalerate, therapeutic formulations and uses thereof |
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WO2008120778A1 (ja) | 2008-10-09 |
US20100210726A1 (en) | 2010-08-19 |
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EP2143434A1 (en) | 2010-01-13 |
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