JP5268362B2 - 寿命を拡大させ、かつ加齢関連疾患の発症を遅らせるための方法 - Google Patents
寿命を拡大させ、かつ加齢関連疾患の発症を遅らせるための方法 Download PDFInfo
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Description
本発明は、生物における寿命を拡大させて、癌を含む加齢関連疾患に関連した合併症の発症および多くを遅らせる方法に関する。より詳しくは、本発明は、カロリー制限において活性化される場合と同じ有益な遺伝子の発現(すなわち遺伝子の活性化)をアップレギュレートおよびダウンレギュレートするために、化学物質を投与することに関する。遺伝子は、カロリー摂取を低減する必要なく、カロリー制限下において認められる条件と同じ細胞内条件を模倣することによって活性化される。寿命を延長させて、生物を加齢関連疾患から保護するための組成物および方法も同様に提供される。
単細胞生物および多細胞生物における寿命を拡大させるために多くの試みがなされている。これらの試みには、様々な栄養に基づく介入、ビタミンサプリメント、抗酸化剤サプリメント、運動、ホルモン的、薬学的および他の範例が含まれている(Lane, M. et al. Nutritional Modulation of aging in nonhuman primates, 1999 The Journal of Nutrition, Health & Aging, Vol. 3, No. 2 pp 69-76)。これらの試みによって時に、よりよい健康が得られるが、過去70年間において有益な遺伝子の活性化のみが寿命の増加を引き起こした。これらの有益な遺伝子の活性化法は、平均寿命および最大寿命を拡大させることが証明されている:1)カロリー制限(CR)による遺伝子活性化;2)遺伝子操作(遺伝子の人工的な付加または欠失)を受ける動物の特定のタイプ;および3)共基質NAD+[24]に関するSir-2酵素のミカエリス定数Kmを低下させることによってSir2遺伝子を活性化する化学物質の使用。CRは、糖質、脂質、またはタンパク質に由来する総カロリーを、自由に食餌を与えた対照動物のカロリーの25%〜60%低いレベルまで制限することである(Koubova et al, How does calorie restriction work? 2003 Genes & Development. Vol. 17 pp 212-221)。CRによる遺伝子活性化によって寿命を拡大させることの成功には、酵母、輪形動物、グッピー、クモ、ショウジョウバエ、ハムスター、ラット、マウスを含む広範囲の異なる生物が含まれ、現在、霊長類において寿命を拡大させることが示されている(Lane et al.;Koubova et al.;Lane et al, Short-term calorie restriction improves disease-related markers in older male rhesus monkeys (Macaca mulatta) 1999 Mechanisms of Ageing and Development Vol. 112 pp 185-196)。遺伝子操作によって寿命を拡大させることの成功は、酵母、ぜん虫、ショウジョウバエ、およびマウスにおいて成功している(Hekimi, S. et al, Genetics and the Specificity of the Aging Process, Science. 2003 Feb 28;299(5611):1351-4. Review; Guarente, L. SIR2 and aging- the exception that proves the rule, Trends Genet. 2001 Jul;17(7):391-2; Tissenbaum, H et al. Increased dosage of a sir-2 gene extends lifespan in Caenorhabditis elegans, 2001 Nature VoI 410 pp 227-230; Lin, S et al, Requirement of NAD and SIR2 for Life-Span Extension by Calorie Restriction in Saccharomyces cerevisiae 2000 Science Vol. 289 pp 294-297; Lin, S et al, Calorie restriction extends Saccharomyces cerevisiae lifespan by increasing respiration 2002 Nature, Vol. 110 pp 244-248; Guarente, L. Mutant mice live longer, 1999 Nature Vol. 402 pp 243-245)。NADに関するSir-2酵素のミカエリス定数を低下させる化学物質による寿命の延長の成功は、酵母およびぜん虫において示されている(Howitz et al., Small molecule activators of sirtuins extend Saccharomyces cerevisiae lifespan, Nature 425: 191-196; Wood et al., Sirtuin activators mimic caloric restriction and delay ageing in metazoans, Nature, Volume 430, 5 August 2004)。全ての場合において、寿命の拡大は有益な遺伝子の活性化を必要とした。
本発明の開示は、寿命を拡大させるための、およびそれを必要とする個体における加齢関連疾患の発症を遅らせるための方法および組成物に関する。本発明の一つの局面において、生物の寿命を拡大させるための方法が提供される。方法には、オキサロアセテート、オキサロ酢酸、オキサロアセテート塩、またはその代謝的前駆体であるα-ケトグルタレートまたはアスパルテートのような化合物の有効量を投与する段階が含まれる。化合物は、経口、局所、および/または非経口投与することができる。化合物は緩衝剤と共に調剤されることが都合がよい。任意で、生物は哺乳動物である。本発明の一つの局面において、哺乳動物はヒトである。
本発明は、それを必要とする個体における寿命を拡大させるための、および加齢に関連した障害を処置するための組成物および方法に関する。本発明は、オキサロアセテート、ならびにα-ケトグルタレートおよびアスパルテートを含む化学前駆体の投与によって、線虫C.エレガンス(C. elegans)のような単純な動物、より複雑なD.メラノガスター(D. melanogaster)および複雑な哺乳動物を含む多細胞対照生物の類似の集団と比較して、平均集団寿命の36%までの増加、および最高寿命の40%までの用量依存的な寿命の増加が得られる、という驚くべき発見に一部基づいている。特定の理論に拘束されることなく、オキサロアセテート化合物またはその前駆体との細胞外接触およびオキサロアセテートのその後の細胞内への転移によって、生物の寿命を増加させる有益な遺伝子を活性化させる代謝的シグナル伝達の変化が起こると考えられている。本明細書において用いられるように、「オキサロアセテート」という用語には、オキサロアセテート、その塩、ならびにα-ケトグルタレートおよびアスパルテートが含まれるがそれらに限定されるわけではないオキサロアセテートの化学前駆体が含まれる。「それを必要とする個体」という句は、オキサロアセテートの寿命拡大および/または抗加齢効果から恩恵を受けるであろう任意の多細胞生物を指す。個体には、オキサロアセテートの投与に対して感受性がある任意の脊椎動物または無脊椎動物が含まれるがそれらに限定されるわけではない。例としての脊椎動物には、魚類、両生類、は虫類、鳥類、およびヒト、霊長類、イヌ、ネコ、または他の動物のような哺乳動物が含まれる。
オキサロアセテートは、寿命を延長させるため、および/または加齢関連疾患および体重障害を改善するために、治療的有効量で個体に投与することができる。本明細書において用いられるように、「オキサロアセテート」には、オキサロ酢酸、酸の塩、またはオキサロアセテートの緩衝溶液、ならびにそれらの混合物が含まれる。この用語には同様に、α-ケトグルタレートおよびアスパルテートのようなオキサロアセテートの前駆体が含まれる。
オキサロアセテートの毒性および治療効力は、細胞培養または実験動物において、標準的な薬学的技法によって、例えばLD50(集団の50%に対して致死的である用量)およびED50(集団の50%において治療的に有効である用量)を決定することによって決定することができる。毒性対治療効果の用量比は治療指数であり、これは比LD50/ED50として表記されうる。マウスに関するα-ケトグルタレートのLD50は、5 g/kg体重より上である。オキサロアセテートのLD50は、5 g/kg体重より上である。オキサロアセテートは、あらゆる細胞のクエン酸サイクルに関係する化学物質から予想されるように、非常に毒性が低い。
「Studies on Anti-diabetic Effect of Sodium Oxaloacetate」, Tohoku Journal of Experimental Medicine, 1968, volume 96, pp. 127-141)。ヒトにおいて提唱されているより300%〜50,000%高い用量でマウスにおいてオキサロアセテートを用いた本発明者らによるさらなる試験は、毒性が陰性であることを示しており、マウスは対照群より長く生存する。妥当な量における本化合物の安全性は、化合物がヒト代謝物であることから保証される。
本発明に従って用いるための薬学的組成物は、一つまたは複数の生理的に許容される担体または賦形剤を用いて通常のように調剤してもよい。このように、オキサロアセテートおよびその生理的に許容される塩および溶媒和化合物は、吸入もしくは吹送(口または鼻のいずれかを通して)による投与、または経口、口腔内、局所、経皮、非経口、もしくは直腸内投与のために調剤されてもよい。吸入の場合、オキサロアセテートの投与は、たとえ投与されたオキサロアセテートの用量が、生物全体に恩恵を与えるために必要な用量より少ない場合であっても、肺組織に直接加齢恩恵を提供するであろう。オキサロアセテートの吸入は、肺の加齢関連疾患の発症を遅らせて、肺疾患からの保護を提供するであろう。
本明細書の特定の局面は、以下の実施例を参照することによってより容易に理解されうるが、これらの実施例は本明細書の教示を例示することを意図しており、特定の例示された態様にその範囲を制限しない。
シノラブディス・エレガンス線虫の遺伝子学は十分に理解され、基礎細胞エネルギー経路がヒトを含む全ての動物種を通して良好に保存されていることから、シノラブディス・エレガンス線虫が試験動物として一般的に用いられる。代謝エネルギー経路に関するこれらの十分に理解された虫による試験は、動物界における試験である。エネルギー経路の保存は、CRが、調べた全ての動物種において作用する理由である。
実施例1における実験を、線虫成長寒天においてオキサロアセテートの濃度範囲に関して繰り返した。C.エレガンスに関する寿命の増加は、寒天における2 mMオキサロアセテートで認められ、4 mM、6 mM、8 mM、10 mM、12 mM、14 mM、および16 mM濃度で増加した。寿命の最高の増加は、調べた濃度に関して16 mM(約25%)であり、最低の増加は2 mM(約10%)であった。
スプリットマイシン単独と寒天、ならびにスプリットマイシンとオキサロアセテートおよび寒天を含むさらなるプレートも同様に用いたことを除き、実施例1における実験を繰り返した。スプリットマイシンは、Sir2遺伝子(酵母におけるSir2a、ヒトにおけるSirt1)に関する選択的阻害剤である。Sir2機能を阻害したがオキサロアセテートを含まないプレート上の線虫は、図3において反映されるように対照群より生存期間が短かった。図3は、CRと同様に、オキサロアセテートがSilent Information Regulator遺伝子(Sir2)をアップレギュレートして寿命を増加させることを図示している。これは同様に、他の有益なCR活性化遺伝子がオキサロアセテートによってアップレギュレートおよびダウンレギュレートされて、Sir2活性化がなくとも寿命を増加させることを示している。オキサロアセテートを含むプレート上の線虫は、対照群より約36%長く生存した。Sir2機能が阻害されたがオキサロアセテートを含むプレート上の線虫は、対照群より15%の寿命の増加を有した。このことは、Sir2がCRにおける寿命の増加の約1/3を構成するが、他のCR有益遺伝子が、寿命の増加の2/3もの多くに寄与することを証明している。これはまた、オキサロアセテートの添加によって活性化された遺伝子の一つがSir2(酵母におけるSir2a、ヒトにおけるSirt1)であることを証明する。
実施例4における実験を、フードバイアルを3〜4日毎に交換しなかったこと、およびハエ71匹にオキサロアセテートを補給してハエ70匹には補給しなかったことを除き、繰り返した。これによって、ハエを攻撃する微生物病原体の外寄生が起こり、ハエを「ストレス」下に置いた。カロリー制限動物は、自由に飼料を与えた対照動物より良好にストレスから生存することは文献において周知である。実験からのデータを図表にして、図5に表す。
約9ヶ月齢のC57Bl/6系雄性マウスを、Harlan, San Diego, CAから役目を終えた繁殖動物として得た。これらのマウスは全て同じ日に生まれた。C57Bl/6系マウスは、個体間の遺伝子変動を低減させるための近交系である。マウスを個別ケージに収容して、自由な量の栄養強化フードペレット(Kadee)を提供した。摂取された食物量の重量を毎日測定して、マウスの体重を毎週測定した。2週間後、マウスによって摂取された平均食物量のベースラインはマウス1日あたり7.1 gであると確立された。さらに、一晩絶食後、各マウスの血中グルコースの値を各マウスに関して記録した。2週目の時点でマウスを等しい三つの群(初回体重および絶食時血糖測定に基づいて)、I)自由に飼料を与える「対照群」8匹;II)「カロリー制限」マウス8匹、最初は対照群より20%少ない飼料を2週間与え、その後対照群より40%少ない量を与える;およびIII)その飼料中にオキサロアセテートの増加量を与えたが自由に餌をとらせた「オキサロアセテート」補給マウス9匹、に分けた。
実施例6における実験を1ヶ月齢の雄性マウスについて繰り返した(実施例6において用いたマウスが9ヶ月齢であるのに対し)。先の実施例6の場合のように、若いC57Bl/6系マウスを体重によって三群に分けた:I)対照;II)カロリー制限;およびIII)オキサロアセテート補給。しかし、オキサロアセテートの用量を徐々に増加させる代わりに、実験開始からマウスに濃度0.4%のオキサロアセテートを供して、カロリー制限マウスには40%制限食を直ちに与えた。マウスの体重を毎週測定して、結果を図7に表されるようにプロットした。
実施例6のC57Bl/6マウスの3群のそれぞれのマウス3匹を21週間後に屠殺した。各動物の肝臓を抽出して、マウス3匹のそれぞれの組をプールして、Caoらによって記述されるように遺伝子活性に関して分析した。プールしたマウスの群の遺伝子発現は、オキサロアセテートの補給に反応して、対照群のマウスと比較してCR下のマウスと同様に変化することが示された。CRマウスは、遺伝子1,763個の発現において測定可能な変化を示したのに対し、オキサロアセテートマウスは遺伝子765個において測定可能な変化を示した。各群はマウス3匹のプールであることから、個々の反応には変動がある。群の身体の結果(体重減少、健康期間の増加、および寿命の増加)の変化は、群によって共通に発現される遺伝子により、個体間の変動ではない。このように、対照群から発現が変化したが、カロリー制限群とオキサロアセテート補給群の間で共通に発現される遺伝子を観察することが重要である。オキサロアセテート群とカロリー制限群で共通に発現され、対照群と比較してゲノム発現が変化した遺伝子は363個であった。これらの共通の遺伝子363個の方向性分析から、オキサロアセテート群のマウスとカロリー制限群のマウスで共通に発現された変化した遺伝子の98%が、対照群のマウスと比較して同じ方向に移動する(アップレギュレートまたはダウンレギュレート)ことが示された。このように、オキサロアセテートの補給は、カロリー制限下のマウスと同じゲノム変化を模倣し、遺伝子の98%が共通に対照群から発現が変化した。しかし、オキサロアセテートマウスは、その飼料を制限する必要はなく、自由に餌を食べた。オキサロアセテート摂取マウスとカロリー制限マウスとの変化における98%類似性はまた、オキサロアセテート補給マウスが対照マウスほど多くの体重獲得を示さず(先の実施例6および7)、より長生きしてより健康であった(以下の実施例9)という事実と結びつく。オキサロアセテート補給はカロリー制限と同じ不可欠なゲノム反応を模倣する(図8を参照されたい)。C.エレガンスおよびD.メラノガスターに与えたオキサロアセテートの有効量は、寿命を20〜36%増加させて、マウスの寿命の最小の増加は10%である。
実施例8において屠殺されなかった実施例6のC57Bl/6マウスを異なる三群として飼料を与え続けた:1)対照、2)カロリー制限、および3)オキサロアセテート補給(持続用量として飼料中に0.4%/重量)。カロリー制限群およびオキサロアセテート群は実験を通して最も健康な群であることが認められたが、対照群の一部は、間欠性の痒みの発作および皮膚の発赤を特徴とする炎症に罹患していた。本実験は特許のために本出願が提出された際に継続中であるが、オキサロアセテート群は対照群より長く生きて、カロリー制限群と少なくとも等しいことが最初に示されている。
体重約200 kgの肥満個体を同定して、オキサロアセテート1,000 mgを30日間1日おきに経口投与する。個体はカロリー制限を受けていない。体脂肪および体重の低減が処置後に観察される。
心疾患および骨粗鬆症を含む加齢関連障害を示す個体を同定する。個体の半数にオキサロアセテート約7 mg/kg体重の有効量を1日毎に30日間またはそれより多く投与する。残りの半数の個体にはプラセボを投与する。オキサロアセテートを投与された個体は、絶食時血糖値、絶食時インスリンレベル、トリグリセリド、Hs-CRPレベル、総コレステロール、LDLコレステロール、ならびに収縮期および拡張期圧の低減を含む、加齢に関連した症状の低減を示す。さらに、無処置個体と比較してアテローム性動脈硬化症のリスクの低減も同様に観察される。
2群の個体を太陽からのUV照射に曝露した。1群は、濃度8mMのオキサロアセテートを皮膚に塗布したが、対照群は塗布しない。両群の皮膚を、DNA修復の相対速度を測定するために、予定されていないDNA合成(UDS)に関して測定した。オキサロアセテートを有する群は、対照群よりUDSの有意な増加を有する。
2群の個体が、類似のタイプの癌腫瘍を有すると診断される。1群は、オキサロアセテート2,000 mg/日の用量のオキサロアセテートを服用し、対照群は服用しない。オキサロアセテートは、腫瘍が利用できるグルコース量を低減させ、カロリー制限において認められる方法と同様に腫瘍の成長を制限する。対照群は腫瘍の成長に制限を認めない。
Claims (7)
- 生物の寿命を拡大させるための薬剤を製造するための、オキサロアセテート、オキサロ酢酸、およびオキサロアセテート塩からなる群より選択される化合物の有効な寿命拡大量の使用。
- 化合物が経口投与のために調剤される、請求項1記載の使用。
- 化合物が緩衝剤と共に調剤される、請求項1記載の使用。
- 生物が哺乳動物である、請求項1記載の使用。
- 哺乳動物がヒトである、請求項4記載の使用。
- 薬剤が局所投与される、請求項1記載の使用。
- 薬剤が非経口投与される、請求項1記載の使用。
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JP2013005607A Active JP5787911B2 (ja) | 2004-12-17 | 2013-01-16 | 寿命を拡大させ、かつ加齢関連疾患の発症を遅らせるための方法 |
JP2015093124A Withdrawn JP2015145417A (ja) | 2004-12-17 | 2015-04-30 | 寿命を拡大させ、かつ加齢関連疾患の発症を遅らせるための方法 |
JP2016206504A Pending JP2017019864A (ja) | 2004-12-17 | 2016-10-21 | 寿命を拡大させ、かつ加齢関連疾患の発症を遅らせるための方法 |
JP2018096719A Active JP6865193B2 (ja) | 2004-12-17 | 2018-05-21 | 寿命を拡大させ、かつ加齢関連疾患の発症を遅らせるための方法 |
JP2020013377A Pending JP2020075936A (ja) | 2004-12-17 | 2020-01-30 | 寿命を拡大させ、かつ加齢関連疾患の発症を遅らせるための方法 |
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JP2015093124A Withdrawn JP2015145417A (ja) | 2004-12-17 | 2015-04-30 | 寿命を拡大させ、かつ加齢関連疾患の発症を遅らせるための方法 |
JP2016206504A Pending JP2017019864A (ja) | 2004-12-17 | 2016-10-21 | 寿命を拡大させ、かつ加齢関連疾患の発症を遅らせるための方法 |
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JP2020013377A Pending JP2020075936A (ja) | 2004-12-17 | 2020-01-30 | 寿命を拡大させ、かつ加齢関連疾患の発症を遅らせるための方法 |
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Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2017019864A (ja) * | 2004-12-17 | 2017-01-26 | アラン ビー. キャッシュ | 寿命を拡大させ、かつ加齢関連疾患の発症を遅らせるための方法 |
WO2018057737A1 (en) | 2016-09-22 | 2018-03-29 | Cash Alan B | Method to alleviate the symptoms of pms |
US11071722B2 (en) | 2016-09-22 | 2021-07-27 | Alan B. Cash | Method to alleviate the symptoms of PMS |
US11865092B2 (en) | 2016-09-22 | 2024-01-09 | Alan B. Cash | Method to alleviate the symptoms of PMS |
Also Published As
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JP5787911B2 (ja) | 2015-09-30 |
US10016385B2 (en) | 2018-07-10 |
WO2006066244A2 (en) | 2006-06-22 |
US20190008816A1 (en) | 2019-01-10 |
JP2017019864A (ja) | 2017-01-26 |
JP2013100323A (ja) | 2013-05-23 |
CA2589995A1 (en) | 2006-06-22 |
EP1824470B1 (en) | 2014-08-06 |
ES2510690T3 (es) | 2014-10-21 |
JP6865193B2 (ja) | 2021-04-28 |
AU2005316295B2 (en) | 2012-03-22 |
WO2006066244A3 (en) | 2007-02-22 |
EP1824470A4 (en) | 2009-07-01 |
CA2589995C (en) | 2016-01-05 |
EP1824470A2 (en) | 2007-08-29 |
CA2911468C (en) | 2018-04-24 |
CA2911468A1 (en) | 2006-06-22 |
JP2008524256A (ja) | 2008-07-10 |
AU2005316295A1 (en) | 2006-06-22 |
JP2015145417A (ja) | 2015-08-13 |
JP2020075936A (ja) | 2020-05-21 |
US11173139B2 (en) | 2021-11-16 |
JP2018131457A (ja) | 2018-08-23 |
DK1824470T3 (da) | 2014-08-18 |
US20080279786A1 (en) | 2008-11-13 |
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