JP6062632B2 - インビボにおける栄養素持続放出 - Google Patents
インビボにおける栄養素持続放出 Download PDFInfo
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- JP6062632B2 JP6062632B2 JP2011542457A JP2011542457A JP6062632B2 JP 6062632 B2 JP6062632 B2 JP 6062632B2 JP 2011542457 A JP2011542457 A JP 2011542457A JP 2011542457 A JP2011542457 A JP 2011542457A JP 6062632 B2 JP6062632 B2 JP 6062632B2
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Description
本出願は、2008年12月17日に出願された国際特許出願PCT/US2008/087104、及び2008年12月17日に出願された米国特許出願第12/337,022号の優先権を主張するものであり、これら両出願は、2007年12月17日に出願された米国特許仮出願第61/014,251号の優先権の利益を主張している。
本発明は、炭水化物、アミノ酸、及び電解質を含む栄養素の適切に時間指定された放出及び増大された吸収を提供することによる、運動選手のパフォーマンスを増大するための組成物に関する。
スポーツに関連した栄養要求の最新の理解は、炭水化物消費は、運動選手の持久力及びパフォーマンスの向上において重要な役割を果たすことを指摘している。1960年代半ばに始まったGATORADE(登録商標)及びその他の類似のスポーツ飲料を入口とする、運動イベントにおけるヒトパフォーマンスに対する様々な栄養素の役割の理解に、より注目が集まっている。
本発明者らは、驚くべきことにGATORADE(登録商標)などの従来型食品と比べ、上昇されかつ延長された血糖反応を提供する、インビボ投与のための組成物を発見した。本発明は、運動パフォーマンスは、栄養補給剤(炭水化物及び電解質など)の制御放出を提供することにより、改善され得るという前提を基にしている。
以下の詳細な説明は、例として示されているが、説明された具体的実施態様のみに本発明を限定することは意図されず、下記の添付図面と一緒に、理解を最良にすることができる。
本発明の様々な実施態様の実行及び使用は以下に詳細に説明されているが、本発明は、多種多様な具体的状況で実施され得る多くの適用可能な発明の概念を提供することは理解されなければならない。本明細書において説明された具体的実施態様は、本発明の実行及び使用のための具体的方式の単なる例証であり、かつ本発明の範囲又はそれに添付された請求項の範囲の限界を定めるものではない。
本実施例の目的は、ピーク運動パフォーマンスを促進する様式で、運動選手へ炭水化物及び他の栄養素を送達する栄養液(nutrition fluid)を開発することである。提唱されたアプローチは、好適なマイクロスフェア又はナノスフェアの水性分散液から可消化炭水化物の制御放出を操作することである。重要な可消化炭水化物は、単糖−ブドウ糖、果糖及びガラクトース;二糖−ショ糖、麦芽糖及び乳糖;並びに、多糖−デンプンを含む。デンプンは、唾液アミラーゼ(口中)及び膵アミラーゼ(小腸中)により、デキストリンへ分解される。デキストリンは、小腸の刷子縁酵素により作用を受け、これも二糖類を単糖類へ変換する。これらの単糖は最終的には、腸上皮を超えて血流へと輸送される。提唱された研究は、血液への持続した取り込みのために、可消化炭水化物、特に単糖類、ブドウ糖及び果糖の制御放出を探求している。
疎水化された多糖は、水性環境中でのそれらの自己集合特性のために、ナノ粒子の合成を高度に促進する。Akiyoshi及びSunamotoは、コレステロールなどの疎水性物質により官能基化された多糖は、水中に分散された場合に、自発的にナノ粒子を形成したことを認めた(Akiyoshi, K.;Sunamoto, J.の論文、「疎水化された多糖の超分子集合体(Supramolecular assembly of hydrophobized polysaccharides)」、Supramolecular Science, 1996, 3, 157-163)。このナノ粒子のサイズ、密度及びコロイド安定性は、疎水性物質のグラフト密度及び疎水度を調整することにより制御することができる。プルラン、デキストラン及びマンナンなどの多糖は、アルキル長鎖及びコレステロールなどの様々な疎水基により部分的に置換された。例えば分子量55kDaのプルランは、コレステロールにより官能基化される場合(〜1.7コレステロール部分/ブドウ糖100ユニット)、サイズが20〜30nmであるナノ粒子を自発的に形成した(Akiyoshi, K.;Deguchi, S.;Tajima, H.;Nishikawa, T.;Sunamota, J.の論文、「疎水化された多糖の自己集合体:ヒドロゲルナノ粒子の構造及び有機化合物との複合体形成(Self-assembly of hydrophobized polysaccharide: Structure of hydrogel nanoparticle and complexation with organic compounds)」、Proc. Japan Acad. 1995, 71, 15-19)。コレステロールを有するプルランは、自己凝集され、水中の懸濁液の超音波処理後に、単分散の安定したナノ粒子を形成した。90℃で1時間加熱した後であっても、凝固は生じなかった。これらのナノ粒子は、抗腫瘍アドリアマイシンなどの疎水性物質(Akiyoshi, K.;Taniguchi, I;Fukui, H.;Sunamoto, J.の論文、「疎水化された多糖の自己集合により形成されたヒドロゲルナノ粒子:複合体形成によるアドリアマイシンの安定化(Hydrogel nanoparticle formed by self- assembly of hydrophobized polysaccharide. Stabilization of adriamycin by complexation)」、European Journal of Pharmaceutics and Biopharmaceutics, 1996, 42, 286-290)、及び様々な水溶性タンパク質の受入れのために使用されるが、小型の水溶性分子の封入は報告されなかった。
更に先に詳述された本発明の様々な態様に対し、エネルギー源として比較的重要な炭水化物(CHO)の基本的理解は、本発明の送達システムのデザインにおいて有用である。運動選手の持久能及び運動パフォーマンスを向上することができる食品製剤は、スポーツ科学及び運動科学におけるいくつかの研究の中心となっているi。運動選手は、疲労を避けるために、運動期間中に燃料の連続供給を必要としている。タンパク質、脂肪及び炭水化物は全て、運動選手の食事の重要な成分として考えられてきたがii,iii、運動選手にとって炭水化物-豊富な食事の現在の強調は、約90年前に行われた研究を基にしている。これらの初期の研究は、血中ブドウ糖濃度と疲労の間の関係を示唆した。Levineらは1924年に、25マイルマラソンレースの参加者への炭水化物-豊富な食事は、向上したランニングパフォーマンス(レースを完走する時間として測定)を生じ、かつランナーにおける低血糖を防止したことを発見したiv。他方でChristensenとHansenは、運動時の高脂肪食は、低血糖及び神経低糖症を引き起こし、これは運動後の重篤な疲労及び消耗を引き起こしたことを示したv,vi,vii。彼等は、これらの症状は、運動前に炭水化物食を用いることにより、予防することができることを発見した。
ヒドロキシプロピルセルロース(HPC)などの温度反応性ポリマーのマイクロ粒子を、このポリマーの水溶液をその下限臨界溶解温度以上に加熱することにより調製した。これらの粒子内のポリマー鎖を、FDA承認のトリメタリン酸三ナトリウム(TSTMP)を用い、共有結合架橋し、マイクロ粒子ヒドロゲルを得た。これらの粒子を、デキストロース(D-ブドウ糖)で負荷し、捕獲されたデキストロースの放出速度を、様々な化学組成及び粒子濃度を持つ製剤について試験した。水-膨潤性ヒドロゲル粒子内に存在する糖質は、遅延放出に利用可能であった。残りの糖質は、水相に存在し、腸管腔を超える即時吸収に利用可能であった。これらのヒドロゲルマイクロ粒子を、アルギン酸ナトリウムなどのpH反応性で粘膜付着性のポリマーによりコーティングし、胃放出に対する拡散障壁を提供した。インビトロ放出動態及びインビボ放出動態の両方(2つの異なるエネルギー消費速度で)を、実験により決定した。本発明の遅延放出型製剤に関するブドウ糖濃度、対、時間プロファイルは、市販の従来型即時放出型製剤及び他の対照に対する差異及びこれらに勝る利点を明確に示した。
ヒドロキシプロピルセルロース(HPC-SL、USP等級)は、Nippon Soda社から入手した。精製大豆レシチンは、MP Biomedicals社(カタログ番号102147)から購入した。アルギン酸ナトリウムポリマー(アルギン酸ナトリウムNF、F-200、SAHMUP、及びアルギン酸ナトリウムNF、SALMUP)は、American International Chemical社から入手した。トリメタリン酸三ナトリウム(TSTMP、試薬等級)及び水酸化ナトリウム(試薬等級、>98%)は、Sigma-Aldrich社から購入した。グルコースオキシダーゼ/ペルオキシダーゼ酵素(PGO酵素カプセル、製品番号P7119)、o-ジアニシジン二塩酸塩(カタログ番号D3252)、デキストロース(カタログ番号D9434)及び塩酸(37%、カタログ番号320331)は、Sigma-Aldrich社から入手した。Thin-N-Thik(登録商標)99デンプン及びResista(登録商標)682デンプン、無水クエン酸、Staleydex(登録商標)333デキストロース、及びKrystar(登録商標)300結晶果糖は、Tate&Lyle社から入手した。食品等級大豆レシチン、UltraLec(登録商標)P脱油レシチンは、Archer Daniels Midland社から入手した。食品等級界面活性剤モノグリセリドのジアセチル酒石酸エステル(DATEM、Panodan(登録商標)150 LP K-A)は、Danisco社から入手した。水酸化ナトリウム(FCC等級)は、VWR社から購入した。安息香酸ナトリウム(FCC等級)は、Fischer Scientific社から購入した。食品等級のソルビン酸カリウム及びトリメタリン酸三ナトリウムは、Spectrum Chemical Mfg社から購入した。全ての化学物質は、更に精製することなく使用した。広く使用された市販のスポーツドリンクであるGATORADE(登録商標)を、インビボ実験の陽性対照として使用した。GATORADE(登録商標)は、水、高果糖コーンシロップ(ブドウ糖-果糖シロップ)、ショ糖シロップ、クエン酸、天然香料、塩分、クエン酸ナトリウム、リン酸一カリウム、食用改質デンプン、赤色色素#40、及びロジンのグリセロールエステルからなる。総糖質濃度は、5.83%(w/v)である。ナトリウム及びカリウム濃度は、各々、0.45mg/mL及び0.125mg/mLである。
ヒドロキシプロピルセルロースは、温度反応性ポリマーである。このポリマー溶液の下限臨界溶解温度(LCST)以上に加熱した場合、水和されたポリマー鎖は、ポリマー−水の水素結合の熱破壊のために、水分を喪失する。このポリマー鎖は、疎水性になり始めるので、溶液から沈殿し、マイクロ粒子を形成する。疎水性相互作用による粒子形成は、可逆性であり−このポリマー分子は、この分散液がLCST以下に冷却された場合に、再度溶解し始める。HPC水溶液の下限臨界溶解温度に対する様々な添加剤の作用を、示差走査熱量測定を用い測定した。HPC水溶液(8%w/v)のLCSTは、48℃であった。3.2%(w/v)大豆レシチン溶液4mLを、8%(w/v)HPC溶液(10mL)へ添加した場合、LCSTの変化は認められなかった。水中TSTMP溶液(1.77%w/v)3gを、HPC及び大豆レシチンを含有する溶液に添加した場合、LCSTは37℃に低下した。最後に、1.36%w/v水酸化ナトリウム溶液0.5gを添加し、かつこの分散液を、300rpmで攪拌しながら、50℃で1時間加熱した。ポリマー粒子の固形物の沈殿が、1時間の加熱後に認められ、これは室温への冷却後に容易に再分散することができた。この分散液のpHは、4N塩酸40μLを添加することにより、約7に調節した。デキストロース(1.75g)をこの分散液に添加し、かつ攪拌により溶解した。デキストロース添加後の、分散液中の架橋されたHPCのLCSTは、約32℃であった。HPCのLCSTに対する添加剤の作用についてのこれらの測定から、架橋剤を使用しなくとも、粒子の形成が生じることは明らかである。しかし広範なイオン強度、温度及びpH条件にわたり粒子の完全性を維持するためには、化学架橋が望ましい。
反応温度50℃で、HPC鎖は凝集し、マイクロ粒子を形成する。粒子内の個々のポリマー鎖を、図13に示された反応を用い、共有結合架橋した。この反応の終了時に、粒子はバイアルの底に沈降した。しかしこれらは、室温への冷却後、穏やかに攪拌することにより、容易に再分散した。
ブドウ糖は、グルコースオキシダーゼにより、グルコン酸と過酸化水素に酸化される(図14)。過酸化水素は、ペルオキシダーゼの存在下で、o-ジアニシジンと反応し、着色生成物を形成する。450nmで測定された茶色の強度は、当初のブドウ糖濃度に比例する。
LifeScan社から市販されているOneTouch Ultra血中ブドウ糖バイオセンサーを使用し、血中ブドウ糖濃度を決定した。OneTouch Ultra血中ブドウ糖モニタリングシステムは、血液わずか1μL及び濃度分析に5秒を必要とする先進の電気化学バイオセンサー試験紙を使用するXL。この試験紙は、血液を試験紙に自動的に引き寄せるデザインを特徴としている。
1.実験前24時間は激しい運動を避けること。
2.実験前に、一晩、少なくとも10時間は絶食すること。
3.坐位で採血し、血中ブドウ糖濃度を決定するために、従来型ばね押し式使い捨てランセット、及び携帯型ブドウ糖測定器を使用し分析すること。45分間で最大3回、初回測定値を得かつ記録し、ベースラインブドウ糖レベルを確立すること。
4.被験製剤、水(対照)、水性デキストロース溶液又はGATORADE(登録商標)(陽性対照)の400mLをおよそ2分以内に消費すること。
5.坐位で採血し、携帯型ブドウ糖測定器を使用し血中ブドウ糖濃度を分析すること。最初の90分間は5分毎に、及びその後最大240分間は15分毎に、測定値を得かつ記録し、血中ブドウ糖濃度、対、時間プロファイルを得ること。
1.実験前24時間は激しい運動を避けること。
2.実験前に、一晩、少なくとも10時間は絶食すること。
3.45分前には、試験施設に入り、着座し楽にすること。
4.坐位で採血し、血中ブドウ糖濃度の決定を目的として従来型ばね押し式使い捨てランセット、及び携帯型ブドウ糖測定器を使用し分析すること。45分間で最大3回、初回測定値を得かつ記録し、ベースラインブドウ糖レベルを確立すること。
5.ベースラインブドウ糖レベルが確立した後、約5mph(〜60%VO2max)の速度で最大15分間、トレッドミル上を走行すること。所定の時点で、被験者には:短時間走行を停止すること;採血し、血中ブドウ糖レベルを決定するために、従来型ばね押し式使い捨てランセット、及び携帯型ブドウ糖測定器を使用し、分析すること;並びに、ウォーミングアップ期間が完了するまで、走行を直ちに再開すること:が求められた。15分間で最大3回、初回測定値を得かつ記録し、運動ベースラインブドウ糖濃度を確立すること。ウォーミングアップ期間後直ちに、約2分以内で、被験製剤400mLを消費すること。
6.前記ペースで走行を再開し、所定の時点で、被験者には:短時間走行を停止し、採血し、血中ブドウ糖レベルを決定するために、従来型ばね押し式使い捨てランセット、及び携帯型ブドウ糖測定器を使用し、分析し、並びに、運動アームが完了するまで、各試料採取後、走行を直ちに再開すること:が求められた。被験者には、できる限り長く(最大195分間)、予め設定されたペースで走行することが指示された。通常の原則(regular basis)で測定値を得かつ記録し、血中ブドウ糖濃度、対、時間プロファイルを得ること。
ヒドロキシプロピルセルロース(HPC-SL、4g)を、250-mLエーレンマイヤーフラスコ中で、室温で、磁気攪拌機を使用し、蒸留水50gに溶解した。エーレンマイヤーフラスコ内のこのHPC溶液に、大豆レシチンの水溶液を添加し、この混合液を、均質な淡黄色溶液が得られるまで、5分間攪拌した。この溶液に、TSTMP水溶液を、3アリコートで添加し、各添加の間2分間攪拌した。この製剤の濁りは、TSTMPの添加後増大し、これは粒子形成を示している。最後に、水酸化ナトリウム水溶液を添加し、この分散液を5分間攪拌した。この分散液のpHを、pHメーターを用い測定し、約11.6であった。得られた製剤を、ホットプレート(Corning Instruments社、PC 620D)を用い50℃に維持した水浴で、2時間加熱し、磁気攪拌機を用い、300rpmで攪拌した。この反応の間に、固形沈殿物が形成された。エーレンマイヤーフラスコを、水浴から取り外し、室温まで冷却し、この反応時に形成された固形沈殿物が再分散されるまで磁気攪拌機を用いて混合し、均一な均質分散液を形成した。この分散液のpHを測定し、10.9であった。この分散液のpHを、4N HCl溶液(〜30〜50μL)を用いて、7.8に調節した。その後デキストロース粉末(8.95g)を添加し、かつ固形物が溶解するまで、この混合物を室温で5分間攪拌した。この分散液を50℃で20分間再度加熱し、この間300rpmで攪拌した。形成された沈殿を、室温まで冷却後、磁気攪拌機を用いて再分散させた。
8%(w/v)HPC溶液10mLを、ガラスバイアルに入れた。磁気攪拌機(300rpm)及び攪拌棒(長さ5mm×直径2mm)を使用し攪拌しながら、大豆レシチンの水溶液をこのバイアルに添加した。この溶液に、攪拌を続けながら、TSTMP水溶液を滴加した。最後に、NaOH溶液0.5mLを添加し、得られた溶液を50℃で1時間加熱した。1時間後、この混合物を冷却した。白色固形物が溶液の底に沈降した。沈降した固相中のマイクロ粒子を、200rpm(室温で約1時間)混合することにより、再分散させ、透明で均質な溶液を生じた。こうして得られた分散液を、4N HCl溶液数μLで中和した。この分散液の最終HPC濃度は、約4.4%(w/v)であった。デキストロース粉末(1.85g)を、4.4%(w/v)HPC分散液18.5mLに添加した。この分散液を、デキストロースの溶解が完了するまで、攪拌した。放出試験を行う前に、少なくとも48時間経過した。
疎水性に改質された食用デンプン(Thin-N-Thik(登録商標)99デンプン及びResista(登録商標)682デンプン)などの多糖を使用し、ヒドロゲルマイクロ粒子を形成した。デンプン800mgを蒸留水10mL中に室温で溶解することにより、8%(w/v)デンプン溶液を調製し、混濁しているが均質な分散液を得た。その後精製大豆レシチン、TSTMP、及び水酸化ナトリウムの溶液を、蒸留水中に調製した。ガラスバイアル中に入れたデンプン分散液10mLに、大豆レシチン溶液4mLを添加し、この混合液を5分間攪拌した。この混合液に、TSTMP溶液3mLを添加した。5分間攪拌した後、水酸化ナトリウム溶液0.5mLを添加した。このガラスバイアルを、50℃の油浴中に配置し、60分間加熱した。磁気攪拌機を用い攪拌を提供し、かつ反応の間は速度300rpmを維持した。60分後、バイアルを油浴から取り外し、室温まで冷却し、4M塩酸溶液数μLを使用し、pHを7に調節した。得られる分散液中の粒子サイズを、表5に示した。
下記の製剤を、蒸留水及び食品等級の化学物質を用い、2-リットルガラス反応器内で合成した。アルギン酸ナトリウム(SALMUP)、大豆レシチン、トリメタリン酸三ナトリウム、水酸化ナトリウム、クエン酸、及びデキストロースのストック液を、蒸留水中に調製した。蒸留水250mL中に安息香酸ナトリウム及びソルビン酸カリウムを溶解することにより、安息香酸ナトリウム及びソルビン酸カリウムのストック液を、調製した。容量2-リットルの欧州式先細の3-首のジャケット付きフラスコ(Chemglass社、カタログ番号CG-1576-11)において、反応を実行した。この反応器の内容物は、オーバーヘッド攪拌機(IKA(登録商標)RW-20)、及びテフロン攪拌ブレード(Chemglass社、カタログ番号CG-2080)に装着された磨きガラスシャフト(Chemglass社、カタログ番号CG-2078-02)からなるかき混ぜ機を用いて、混合した。再循環式水浴を使用し、反応器のジャケットを通過する水の温度を制御した。
先に説明された手順を用い、製剤の別のバッチを合成し、4つの部に分けた。これらの各々は、異なる量の糖質デキストロース及び果糖を含んだ。これらの試料の全体の組成、粒子サイズ、及び粘度を、表6に示している。
大豆レシチン、トリメタリン酸三ナトリウム(TSTMP)、水酸化ナトリウム、及びアルギン酸ナトリウムの水溶液を、蒸留水中に各化合物を溶解することにより、各々調製した。無水クエン酸も、蒸留水中に溶解した。水中の安息香酸ナトリウム及びソルビン酸カリウムの溶液も、安息香酸ナトリウム及びソルビン酸カリウムを蒸留水中に溶解することにより、調製した。加えて、水中のデキストロースの溶液を、蒸留水172g中にデキストロース151gを溶解することにより調製した。
製剤の合成は、食品等級の化学物質を用い、1.5-リットルスケールで実行した。大豆レシチン、TSTMP、水酸化ナトリウム、アルギン酸ナトリウム、安息香酸ナトリウム及びソルビン酸カリウムの水溶液を、最初に個別に調製した。無水クエン酸も蒸留水中に溶解させた。
実施例2.1の製剤を使用した。HPC粒子からのデキストロースの放出動態は、pH7、及び温度28℃で測定した。この温度は、分散液中のHPCのLCSTよりも低いので、これを選択した。
アルギン酸ナトリウム(SALMUP)粉末を、実施例2.1に説明された製剤20gに添加した。最終分散液のpHは、4N塩酸を用い、3.8に調節した。この製剤は、8.7%(w/w)デキストロース、3.9%(w/w)HPC、SALMUP、TSTMP、及び大豆レシチンを含有した。同じ製剤を、等質量の水により希釈した。ブドウ糖輸送の動態を、拡散セルを用い28℃で測定した。この希釈した製剤は、4.3%(w/w)デキストロース、1.9%(w/w)HPC、SALMUP、TSTMP、及び大豆レシチンを含有した。ブドウ糖輸送の動態に対する希釈の作用は、実施例2.8に説明された手順に従い、拡散セルを用いて調べた。図17は、当初の製剤と希釈した製剤について、アクセプターコンパートメントのブドウ糖濃度プロファイルを比較した。
HPC粒子からのデキストロースの放出の動態を、pH7及び温度37℃で測定した。この温度は分散液中のHPCのLCSTを上回り、かつヒト体の温度に近いので、これを選択した。実施例2.1の製剤を使用した。約10時間後には、封入されたブドウ糖のほとんど全てが、放出された(図18)。このHPC粒子は、温度がLCSTを上回って上昇した場合(例えば37℃)、親水性状態から疎水性状態への転移を受ける。これらの粒子は縮小されるので、粒子内に存在するブドウ糖分子は、水と共に排出される。HPC分散液が、ブドウ糖(対照)と同じ濃度でブドウ糖溶液の代わりにドナーコンパートメント内に存在する場合、レセプターコンパートメント中のブドウ糖濃度は、より遅い速度で増加することは、図18から明らかである。
HPC粒子からのデキストロース放出の動態を、pH2及び温度37℃で測定した。pH値2は、空腹時の胃液のpHに似ている。実施例2.4の分散液を使用した。この分散液のpHは、4N塩酸を用い、2に調節した。拡散セルのジャケット内の水は、水浴を用い37℃で維持した。このHPC分散液7mLを、ドナーコンパートメントに添加し、酸性水(pH2、塩酸)7mLを、レセプターコンパートメントに添加した。選択された時間間隔で、レセプターコンパートメント内の液体の0.1mLアリコートを採取し、これを等量の蒸留水で置き換えた。採取したアリコートは、容量フラスコを用い10倍希釈し、このブドウ糖濃度を、GM8分析装置(Analox Instruments社)を用いて決定した。図19は、本分散液及びブドウ糖溶液(対照)に関する濃度プロファイルを示している。明らかに、放出速度は、ブドウ糖溶液よりも、HPC分散液の方が遅い。
HPC粒子からのデキストロース放出の動態に対するpHの作用を、実施例2.1の分散液を用い更に例証した。当初の分散液のpHを、4N塩酸を用い、〜7に調節した。アルギン酸ナトリウム(SALMUP)粉末を、この分散液20gに添加し、磁気攪拌機を用いて溶解した。ブドウ糖放出動態の決定(拡散セルを使用する)の数分前に、アルギン酸ナトリウムを含有する分散液約10gを採取し、そのpHを4N塩酸を用い3.8に調節した。この分散液の残り10gのpHを、4N塩酸を用い2に調節した。そのようにして得られた3種の分散液は全て、約8.7%(w/w)デキストロース、3.9%(w/w)HPC、SALMUP、TSTMP、及び大豆レシチンを含有した。28℃でのブドウ糖放出の動態を、実施例2.8において考察したように、拡散セルを用いて決定した。ドナーコンパートメントは、この分散液7mLを含有した。レセプターコンパートメントは、蒸留水7mLを含有し、ドナーコンパートメント中の分散液のpHと同じpHに酸性化した。レセプターコンパートメント内のブドウ糖の濃度は、グルコースオキシダーゼ比色法を用い決定した。図20は、3種の分散液について、レセプターコンパートメントブドウ糖濃度プロファイルを比較した:pH7、3.8、及び2。pH値7と3.8の濃度プロファイルに、ほとんど差異はなかった。しかしこの分散液のpHが、カルボン酸基(アルギン酸塩の)のpKaよりも実質的により低い場合、有意に異なる濃度時間プロファイルが認められた。このデータは、HPC粒子内でブドウ糖分子の拡散障壁を作製するアルギン酸ナトリウムの役割を明確に示している。この障壁特性は、この分散液が消費後に胃環境において遭遇するであろう酸性pHで増強される。
実施例2.4の分散液約12.85oz(380mL、450g)は、一晩絶食後(〜10時間)に消費された。これらの製剤は、米国食品医薬品局(FDA)により「一般に安全と認められる物質(GRAS)」である食品添加物と考えられる物質で構成された。全ての成分の量は、米国FDA及びWHOにより決定された許容レベル内に収まる。HPC分散液の消費の前1時間に、空腹時血中ブドウ糖濃度を測定し、ベースラインを確立した。この血中ブドウ糖濃度は、分散液の消費後規定間隔で決定した(図21a)。図21aは、GATORADE(登録商標)380mLの消費後の、同じ被験者に関する血中ブドウ糖濃度プロファイルも示している。図21bにおいて、血中ブドウ糖濃度は、空腹時ベースラインブドウ糖濃度を減算することにより、標準化されている。HPC分散液及びGATORADE(登録商標)の両方共、高いグリセミック指数(血中ブドウ糖濃度の上昇速度に反映された、摂取された炭水化物が腸管吸収に利用可能になる速度の測定値)を示した。
図22は、実施例2.5のHPC分散液の血中ブドウ糖濃度プロファイルを、GATORADE(登録商標)対照のそれと比較している。本製剤は、米国食品医薬品局(FDA)により「一般に安全と認められる物質(GRAS)」である食品添加物と考えられる物質で構成された。全ての成分の量は、米国FDA及びWHOにより決定された許容レベル内に収まる。等容量(380mL)のこれら2種の製剤を消費した。両方の製剤は、類似した糖質濃度を含んだ。本実施例のHPC分散液は、実施例2.13のHPC分散液中のHPC粒子の総数のほぼ半分を有する。従って、血中ブドウ糖濃度は、わずかに最大でも約75分間、空腹時濃度を上回り維持され、それでもこれは対照よりも約25分間長い。
本実施例は、2つの異なる種類の炭水化物、すなわちデキストロース及び果糖の遅延放出の、血中ブドウ糖濃度プロファイルに対する影響を例証する。果糖は、迅速に筋肉グリコーゲン合成を促進することはできないがviii、これは、インスリン反応を制御し、かつ炭水化物消費後2時間たっても、空腹時値を有意に上回る血中ブドウ糖濃度を維持するために、使用することができる(図23参照)。実施例2.5の2種の異なるHPC分散液(2.5A及び2.5D)を、図23において比較した。一方の分散液は、糖質としてデキストロースのみを含有するのに対し、他方の分散液は、デキストロース及び果糖の両方を含有した。第二の分散液の総糖質濃度は、第一の分散液のそれと同じであった。同じ容量(380mL)の両方の分散液を消費し、血糖反応を測定した。これらの製剤は、米国食品医薬品局(FDA)により「一般に安全と認められる物質(GRAS)」である食品添加物と考えられる物質で構成された。全ての成分の量は、米国FDA及びWHOにより決定された許容レベル内に収まる。図23において、主に果糖を含有する製剤は、予想されたように、より低いグリセミック指数を有することが認められる。この血中ブドウ糖濃度は、最大約230分間、空腹時レベルを上回った。本HPC分散液により提供された遅延放出機構は、果糖のブドウ糖への肝臓での変換において代謝時間のずれがあり、ベースライン値よりも大きい血中ブドウ糖レベルを生じる。
実施例2.7のHPC分散液に関する血中ブドウ糖濃度プロファイルを、対照として使用したブドウ糖の水溶液と比較した(図24参照)。これらの製剤は、米国食品医薬品局(FDA)により「一般に安全と認められる物質(GRAS)」である食品添加物と考えられる物質で構成された。全ての成分の量は、米国FDA及びWHOにより決定された許容レベル内に収まる。これらの製剤は両方共、同様の全体の糖質濃度(〜10重量%)を含んだ。血中ブドウ糖濃度は、ブドウ糖対照(〜60分間)よりも、HPC分散液に関して、より長い期間(〜115分間)空腹時レベルを上回り維持された。このインスリン反応は、即時放出の対照と比べ、遅延放出型製剤に関して明白に同じく有意に低い。
本HPC分散液の摂取のタイミングの作用を、実施例2.7の分散液を用い例証する。本製剤は、米国食品医薬品局(FDA)により「一般に安全と認められる物質(GRAS)」である食品添加物と考えられる物質で構成された。全ての成分の量は、米国FDA及びWHOにより決定された許容レベル内に収まる。本製剤のボーラス380mL(450g)が消費された。このボーラス投与量に関する血中ブドウ糖濃度、対、時間プロファイルを、同量の製剤が3部分で摂取される実験の濃度プロファイルと比較し−これら各部分は、本分散液127mL(150g)からなった。図25から、これらの戦略は両方共、少なくとも最大110分間について、血中ブドウ糖濃度のベースラインレベルを上回ったことが認められた。この血糖反応は、予想されたように、より少量の炭水化物分散液が頻回に消費される場合に、より低かった。
HPC分散液の摂取のタイミングの作用は、対照としてGATORADE(登録商標)を用い、実施例2.7の分散液を用いて例証される。2つの個別の実験において、各製剤380mL(450g)は、3部分で摂取され−各部分は、本製剤127mL(150g)からなった。この血中ブドウ糖濃度は、図26において比較した。
血中ブドウ糖濃度プロファイルに対する中程度の強度の運動(〜60%VO2max)の前に供給された遅延放出型炭水化物の作用を、本実施例において例証している。実施例2.4のHPC分散液380mLを、運動の前に消費した。この実験の対照は、実施例2.4のHPC分散液のデキストロース濃度に近づけるためにデキストロース17.5gを添加したGATORADE(登録商標)製剤380mLであった。図27に認めることができるように、血中ブドウ糖濃度は、HPC分散液において、対照と比べ、50分で有意に高かった。本試験は、被験者が可能な限り長く〜60%VO2maxと等しい速度で走る場合の疲労に対する経過時間の関数としてのエネルギー出力も測定した。HPC分散液は、水対照よりも55%より大きい経過時間、及びデキストロース17.5gが添加された陽性対照GATORADE(登録商標)製剤よりも31%より大きい経過時間を生じた。
本HPC分散液の血糖の影響を、血中ブドウ糖濃度プロファイル曲線下面積を用い、特徴付けた。図28は、実施例2.13から2.19に例示された様々な製剤に関する血中ブドウ糖濃度プロファイルの比較に使用された、濃度時間プロファイルのパラメータを示している。C-Cbは、空腹時(本実験の開始前)の血中ブドウ糖濃度であるベースライン値Cbに対する血中ブドウ糖濃度である。時点t=0は、本実験の開始(例えばCHO製剤の摂取)に対応している。時点t実験は、その間血中ブドウ糖濃度が測定される本実験期間の総時間である。時点tベースラインは、血中ブドウ糖濃度がベースラインを超える、すなわち、空腹時値以下に下落する時点である。血液中のブドウ糖は、CHO摂取後、長期間にわたり、エネルギー源として利用可能であるので、より高いtベースライン値が望ましい。
AUC+は、図28において「+」により示された領域の面積であり、下記式を用い、算出され:
Claims (7)
- 水性懸濁液を含むインビボにおける消費のための組成物であって、
(a) 栄養補給剤;及び
(b) 1以上の(a)の栄養補給剤を封入した1以上のヒドロゲル粒子を含み、
該1以上のヒドロゲル粒子が、
(i) 500ミクロン未満の直径を有するナノ粒子又はマイクロ粒子であり;
(ii) 1以上の化合物であって、共有結合的に架橋され、かつ栄養補給剤のインビボにおける時間制御されかつ持続された放出で、封入された1以上の栄養補給剤を放出する該化合物を含み;
(iii) pH応答性であり、ここで、1以上のヒドロゲル粒子は、胃の酸性環境においては膨潤しないが、小腸へ進入した時点で膨潤し;かつ
(iv) 温度反応性であり、ここで、該ヒドロゲル粒子の1以上の化合物は、水溶液中で下限臨界溶解温度を有し;
ここで、該組成物の1以上の栄養補給剤の炭水化物の放出及び吸収の動態が、該栄養補給剤のインビボにおける時間制御されかつ持続された放出のための化合物を含まない組成物とは異なる、前記組成物。 - 前記ヒドロゲルが、多糖を含む、請求項1記載の組成物。
- 前記多糖が、熱反応性多糖、疎水化多糖、pH反応性多糖、及びそれらの組合せからなる群から選択される、請求項2記載の組成物。
- 前記多糖が、ヒドロキシプロピルセルロースである、請求項3記載の組成物。
- 前記多糖が、アルギン酸ナトリウムである、請求項3記載の組成物。
- 前記組成物が、炭水化物に加えて更に栄養補給剤を含み、該栄養補給剤が、アミノ酸、脂質、電解質、及びビタミンからなる群から選択される、請求項1記載の組成物。
- 前記電解質が、ナトリウム、カリウム、マグネシウム、塩化物、カルシウム、炭酸水素塩、リン酸塩、及び硫酸塩からなる群から選択される、請求項6記載の組成物。
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IL213637B (en) | 2018-07-31 |
KR20110135918A (ko) | 2011-12-20 |
BRPI0922989A2 (pt) | 2015-08-11 |
IL213637A0 (en) | 2011-07-31 |
CN102316752B (zh) | 2014-12-24 |
CA2747659C (en) | 2017-05-23 |
AU2009335743B2 (en) | 2015-10-01 |
WO2010080557A1 (en) | 2010-07-15 |
JP2016144448A (ja) | 2016-08-12 |
US8563066B2 (en) | 2013-10-22 |
CN102316752A (zh) | 2012-01-11 |
BRPI0922989A8 (pt) | 2017-05-23 |
US20090155409A1 (en) | 2009-06-18 |
US20140242212A1 (en) | 2014-08-28 |
AU2009335743A1 (en) | 2011-08-11 |
MX2011006519A (es) | 2011-09-22 |
CA2747659A1 (en) | 2010-07-15 |
US20120015039A1 (en) | 2012-01-19 |
HK1165962A1 (en) | 2012-10-19 |
EP2389075A1 (en) | 2011-11-30 |
KR101791836B1 (ko) | 2017-10-31 |
EP2389075B1 (en) | 2017-11-15 |
US9554586B2 (en) | 2017-01-31 |
US9538775B2 (en) | 2017-01-10 |
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