JP2009102226A - Therapeutic agent for spinal cord injury - Google Patents

Therapeutic agent for spinal cord injury Download PDF

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JP2009102226A
JP2009102226A JP2006037169A JP2006037169A JP2009102226A JP 2009102226 A JP2009102226 A JP 2009102226A JP 2006037169 A JP2006037169 A JP 2006037169A JP 2006037169 A JP2006037169 A JP 2006037169A JP 2009102226 A JP2009102226 A JP 2009102226A
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spinal cord
cord injury
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Chihiro Higashida
千尋 東田
Natsuki Nakayama
なつき 中山
Masashi Yamada
昌司 山田
Hirohito Suzuki
啓仁 鈴木
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Meiji Dairies Corp
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Meiji Milk Products Co Ltd
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Priority to PCT/JP2007/051458 priority patent/WO2007094166A1/en
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    • C—CHEMISTRY; METALLURGY
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    • C07J—STEROIDS
    • C07J17/00—Normal steroids containing carbon, hydrogen, halogen or oxygen, having an oxygen-containing hetero ring not condensed with the cyclopenta(a)hydrophenanthrene skeleton
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    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
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    • A—HUMAN NECESSITIES
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    • A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00

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Abstract

<P>PROBLEM TO BE SOLVED: To provide a drug for recovery from a spinal cord injury recovery exhibiting simple and high effects, to provide a food, a medicine composition containing the same, and to provide a spinal cord injury recovery method. <P>SOLUTION: Provided is Withanoside IV and its peripheral compounds, which are oxygen-containing heterogeneous ring-having steroid glycosides, can be isolated from Ashwagandha (root of Withania somnifera Dunal) and can be synthesized by chemical synthesis means, and have high spinal cord injury-recovering and treating effects. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

本発明は、脊髄損傷治療剤に関し、特にウィタノシド（Ｗｉｔｈａｎｏｓｉｄｅ）ＩＶ及びその周辺化合物を有効成分とする脊髄損傷治療剤、それを含む食品・医薬品組成物、及びそれを用いた脊髄損傷の回復方法に関する。   The present invention relates to a therapeutic agent for spinal cord injury, and particularly relates to a therapeutic agent for spinal cord injury containing withosideside IV and its peripheral compounds, a food / pharmaceutical composition containing the same, and a method for recovering spinal cord injury using the same. .

日本の脊髄損傷患者は１０万人以上となっており、その数は毎年５，０００人ずつ増えていると言われている。脊髄損傷の原因の多くは交通事故、スポーツ事故、高所からの転落・転倒などが挙げられ、これらの受傷時に屈曲、伸展、回旋、圧迫などの外力が複合的に脊椎に作用し、脊椎、脊髄損傷並びに靱帯損傷がもたらされる。（脊椎・脊髄損傷、［平成１８年１月２０日検索］、インターネット＜ｈｔｔｐ：／／ｗｗｗ．ｒｄ．ｍｍｔｒ．ｏｒ．ｊｐ／〜ｓｕｍｉｈｏｓｐ／ｓｅｋｉｓｏｎｂｕｎ．ｈｔｍ＞）。また外傷以外には、脊髄腫傷、椎間板ヘルニアなどがある。   There are over 100,000 people with spinal cord injury in Japan, and the number is said to increase by 5,000 each year. Many causes of spinal cord injury include traffic accidents, sports accidents, falls / falls from high places, etc.When these injuries occur, external forces such as flexion, extension, rotation and compression act on the spine in combination, Spinal cord injury as well as ligament injury results. (Spine and spinal cord injury, [searched on January 20, 2006], Internet <http://www.rd.mmtr.or.jp/-sumihosp/sekisonbun.htm>). In addition to trauma, there are spinal cord injuries and disc herniation.

脊髄が損傷すると、その損傷の仕方、損傷程度、損傷部位により、四肢の運動や感覚の麻痺や、重篤な場合には呼吸障害すら起こる可能性もある。損傷部位の位置が高いほど（仙随→腰随→胸随→頸随）、麻痺の発生する範囲は広く傷害が重度になる。例えば、胸随の損傷であれば下半身麻痺、頸随の損傷であれば全四肢の麻痺となる可能性もある。   If the spinal cord is damaged, depending on how it is damaged, the degree of damage, and the site of damage, limb movement, paralysis of the sensation, and even severe respiratory problems may occur. The higher the location of the injury site (Sengoku → Lumbar → Chest → Neck), the wider the range of paralysis, the more severe the injury. For example, if the chest is damaged, the lower body can be paralyzed, and if the neck is damaged, all limbs can be paralyzed.

脊髄損傷に対する治療法は現在の所、外科的な方法、薬剤投与による方法などが挙げられるが、完全には確立されておらず、また完全損傷の場合には機能回復はかなり困難である。例えば、急性期脊髄損傷に対して現在行われている治療法としては、ステロイド短期大量投与法である（非特許文献１）。しかしこの方法は、受傷後８時間以内に治療を行う必要があり実質困難である。また、ステロイド大量投与法そのものを否定している文献も見受けられ、未だ完全には確立されていないとも言える。また他の方法として、ＥＳ細胞あるいは骨髄間細胞を移植する等の治療法の開発も現在行われている（非特許文献２、非特許文献３）。   Treatment methods for spinal cord injury currently include surgical methods, drug administration methods, etc., but they are not completely established, and in the case of complete injury, functional recovery is quite difficult. For example, a currently used treatment for acute spinal cord injury is a short-term steroid mass administration method (Non-patent Document 1). However, this method is difficult because it requires treatment within 8 hours after injury. In addition, there are documents that deny the steroid mass administration method itself, and it can be said that it has not been completely established. In addition, other methods such as transplantation of ES cells or bone marrow cells are currently being developed (Non-patent Documents 2 and 3).

一方、神経機能不全疾患に対する薬剤による治療法についても種々検討が進んでいる。例えば、特許文献１には脊髄損傷などの神経損傷の治療薬として樹状細胞からの分泌物質や樹状細胞を誘導・増殖・活性化する物質などを有効成分とする治療薬が提案されている。   On the other hand, various studies are also being conducted on therapeutic methods using drugs for neurological dysfunction diseases. For example, Patent Document 1 proposes a therapeutic agent containing a substance secreted from dendritic cells or a substance that induces, proliferates, or activates dendritic cells as a therapeutic agent for nerve damage such as spinal cord injury. .

また、特許文献２、特許文献３、特許文献４では、オキサゾピロロキノリン類、ピロロキノリン類、インドールキノン誘導体などを神経成長因子産出促進剤として提案している。   Patent Document 2, Patent Document 3, and Patent Document 4 propose oxazopyrroloquinolines, pyrroloquinolines, indolequinone derivatives and the like as nerve growth factor production promoters.

クロンバルＥ等（Ｋｒｏｎｖａｌｌ Ｅ． ｅｔ ａｌ．），Ｌａｋａｒｔｉｄｎｉｎｇｅｎ，２００５年６月，１３−２６；１０２（２４−２５）：１８８７−８，１８９０Kronval E. et al., Lakartiddingen, June 2005, 13-26; 102 (24-25): 1887-8, 1890. リロイＰＨ等（Ｌｅｒｏｕ ＰＨ．ｅｔ ａｌ．），Ｂｌｏｏｄ Ｒｅｖ．，２００５年１１月，１９（６），ｐ．３２１−３３１Leroy PH. Et al., Blood Rev. November 2005, 19 (6), p. 321-331 デザワＭ等（Ｄｅｚａｗａ Ｍ． ｅｔ ａｌ．），Ｃｕｒｒ Ｍｏｌ Ｍｅｄ．，２００５年１１月，５（７），ｐ．７２３−７３２Dezawa M. et al., Curr Mol Med. , November 2005, 5 (7), p. 723-732 特開２００４−００２４１２号公報JP 2004-002412 A 特開平０６−００９３９６号公報Japanese Patent Laid-Open No. 06-009396 特開平０６−２１１６６０号公報Japanese Patent Laid-Open No. 06-21660 特開平０７−１１８１５２号公報Japanese Patent Laid-Open No. 07-118152

しかし、上記の特許文献１に示した治療薬及び治療法は樹状細胞あるいはそのサブセットを用いているために簡便性に欠け、規格化も困難である。またそれ以外の上記した方法や治療薬剤の何れについても、未だ完全に確立されたものではなく、その効果についても不十分である。したがって脊髄損傷患者が増加していく背景を考慮すると、脊髄損傷に対して簡便かつ高い効果を示す治療方法及び／又は治療薬剤の開発がより強く望まれている。   However, since the therapeutic agent and the therapeutic method shown in the above-mentioned Patent Document 1 use dendritic cells or a subset thereof, they are not convenient and are difficult to standardize. In addition, any of the other methods and therapeutic agents described above are not completely established yet, and their effects are insufficient. Accordingly, in view of the background of the increase in spinal cord injury patients, development of a therapeutic method and / or therapeutic agent that exhibits a simple and high effect on spinal cord injury is strongly desired.

したがって本発明の目的は、簡便且つ高い効果を示す脊髄損傷を回復する薬剤及び脊髄損傷の回復方法を提供することである。   Therefore, the objective of this invention is providing the chemical | medical agent which recovers the spinal cord injury which shows the simple and high effect, and the recovery method of a spinal cord injury.

上記課題について鋭意検討したところ、本発明者等は脊髄損傷マウスをモデルとして作製し、行動学的評価および免疫組織染色を行った結果、ウィタノシド（Ｗｉｔｈａｎｏｓｉｄｅ）ＩＶ及びその周辺化合物において著しい脊髄損傷回復作用があることを見出し、本発明を完成させた。
すなわち、本発明によれば、ある特定構造を有するウィタノシド（Ｗｉｔｈａｎｏｓｉｄｅ）ＩＶ及びその周辺化合物を提供することで、上記課題を解決することが可能となった。 As a result of intensive studies on the above problems, the present inventors made a spinal cord injury mouse as a model, and performed behavioral evaluation and immunohistochemical staining. As a result, with Witanoside IV and its peripheral compounds, a remarkable spinal cord injury recovery action As a result, the present invention has been completed.
That is, according to the present invention, it is possible to solve the above-mentioned problems by providing withanoside IV having a specific structure and its peripheral compounds.

したがって、本発明のある態様としては、下記構造式１の化合物の少なくとも１種を有効成分とする脊髄損傷治療剤である。   Therefore, as an aspect of the present invention, there is provided a therapeutic agent for spinal cord injury comprising at least one compound represented by the following structural formula 1 as an active ingredient.

構造式１
（式中、Ｒ1及びＲ2はそれぞれ独立して、水素原子、水酸基、アルキル基、またはアルコキシ基であり、Ｒ3は水素原子、水酸基、アルキル基、アルコキシ基、または糖残基である。） Structural formula 1
(In the formula, R 1 and R 2 are each independently a hydrogen atom, a hydroxyl group, an alkyl group, or an alkoxy group, and R 3 is a hydrogen atom, a hydroxyl group, an alkyl group, an alkoxy group, or a sugar residue. )

上記構造式１の脊髄損傷治療剤のうち、特に好ましくは下記構造式２のウィタノシドＩＶを有効成分とする脊髄損傷治療剤である。   Among the therapeutic agents for spinal cord injury of the above structural formula 1, particularly preferred is a therapeutic agent for spinal cord injury containing the vitanoside IV of the following structural formula 2 as an active ingredient.

構造式２
Structural formula 2

また本発明の別の態様としては、上記脊髄損傷治療剤を有効成分として含有し、脊髄損傷の回復作用を有する食品・医薬品組成物である。   Another aspect of the present invention is a food / pharmaceutical composition containing the therapeutic agent for spinal cord injury as an active ingredient and having an action of recovering spinal cord injury.

さらに本発明の別の態様としては、上記脊髄損傷治療剤又は食品・医薬品組成物の有効量を投与することを特徴とする脊髄損傷の回復方法である。   Furthermore, another aspect of the present invention is a method for recovering spinal cord injury, comprising administering an effective amount of the therapeutic agent for spinal cord injury or a food / pharmaceutical composition.

本発明により、簡便かつ高い効果を示す特定の化合物による脊髄損傷治療剤又は食品・医薬品組成物を提供することが可能となる。   According to the present invention, it is possible to provide a therapeutic agent for spinal cord injury or a food / pharmaceutical composition using a specific compound that is simple and highly effective.

また、上記薬剤又はそれの有効量を含む食品・医薬品組成物により、簡便にヒトなどの脊髄治療を目的とした効果的な治療方法を提供することができる。   Moreover, an effective therapeutic method for the purpose of treating spinal cords of humans or the like can be easily provided by the food or pharmaceutical composition containing the drug or an effective amount thereof.

以下、本発明を詳細に説明するが、本発明は以下に述べる個々の形態には限定されない。   Hereinafter, the present invention will be described in detail, but the present invention is not limited to the individual forms described below.

前述の通り上記課題を解決するため、本発明は下記構造式１の化合物の少なくとも１種を有効成分とする脊髄損傷治療剤を提供するものである。   In order to solve the above-described problems as described above, the present invention provides a therapeutic agent for spinal cord injury comprising at least one compound of the following structural formula 1 as an active ingredient.

構造式１
Structural formula 1

上記構造式１において、Ｒ1及びＲ2はそれぞれ独立して、水素原子、水酸基、アルキル基、またはアルコキシ基であることが好ましく、Ｒ3は水素原子、水酸基、アルキル基、アルコキシ基、または糖残基であることが好ましい。Ｒ1及びＲ2は特に好ましくは、それぞれ独立して、水素原子、水酸基のいずれかである。また、Ｒ3は特に好ましくは糖残基である。 In Structural Formula 1, R 1 and R 2 are each independently preferably a hydrogen atom, a hydroxyl group, an alkyl group, or an alkoxy group, and R 3 is a hydrogen atom, a hydroxyl group, an alkyl group, an alkoxy group, or a sugar It is preferably a residue. R 1 and R 2 are particularly preferably each independently a hydrogen atom or a hydroxyl group. R 3 is particularly preferably a sugar residue.

上記置換基としてのアルキル基の例は、本発明はこれらに限定されないが、好ましくは炭素数が１〜６の直鎖状又は分枝状のアルキル基が挙げられ、例えばメチル基、エチル基、プロピル基、イソプロピル基、ブチル基、ペンチル基、ヘキシル基などが挙げられる。また、アルコキシ基の例は、本発明はこれらに限定されないが、好ましくは炭素数が１〜６の直鎖状又は分枝状のアルコキシ基が挙げられ、例えばメトキシ基、エトキシ基、プロポキシ基、イソプロポキシ基、ブトキシ基、イソブトキシ基、ペンチルオキシ基、ヘキシルオキシ基などが挙げられる。   Examples of the alkyl group as the substituent are not limited thereto, but preferably include a linear or branched alkyl group having 1 to 6 carbon atoms, such as a methyl group, an ethyl group, Examples include propyl group, isopropyl group, butyl group, pentyl group, hexyl group and the like. Examples of the alkoxy group include, but are not limited to, the present invention, preferably a linear or branched alkoxy group having 1 to 6 carbon atoms, such as a methoxy group, an ethoxy group, a propoxy group, Examples thereof include an isopropoxy group, a butoxy group, an isobutoxy group, a pentyloxy group, and a hexyloxy group.

上記置換基としての糖残基の例は、本発明はこれらに限定されないが、単糖、糖誘導体、二〜七糖からなるオリゴ糖又はオリゴ糖誘導体等からのグリコシル基が好ましい。結合については、本発明はこれらに限定されないが、α結合、β結合を問わず、またα結合とβ結合が混在していてもよい。具体的な糖残基としては、本発明はこれらに限定されないが、例えばグルコース、ガラクトース、マンノース、フルクトース、フコース、アラビノース等の単糖の残基；グルクロン酸、Ｎ−アセチルグルコサミン、Ｎ−アセチルガラクトサミン、シアル酸等の糖誘導体の残基；マルトース、マルトトリオース、マルトテトラオース、マルトペンタオース、マルトヘキサオース、マルトヘプタオース、ラクトース、ゲンチオビオース、キシロビオース、プリメベロース、メリビオース、ルチノース、ビシアノース、ラミナリビオース、セロビオース、セロトリオース、パノース等の二〜七糖からなるオリゴ糖の残基等が挙げられ、特に好ましくはグルコース及びグルコースで構成される二〜七糖からなるオリゴ糖の残基である。また、２，３，４，６位の水酸基は、例えばアセチル基、マロニル基、リン酸基、メチル基等により修飾されていても良い。   Examples of the sugar residue as the substituent are not limited to these examples, but a glycosyl group from a monosaccharide, a sugar derivative, an oligosaccharide composed of 2 to 7 sugars, an oligosaccharide derivative, or the like is preferable. With respect to the bonds, the present invention is not limited to these, but α bonds and β bonds may be used, and α bonds and β bonds may be mixed. Specific examples of sugar residues include, but are not limited to, the residues of monosaccharides such as glucose, galactose, mannose, fructose, fucose, and arabinose; glucuronic acid, N-acetylglucosamine, N-acetylgalactosamine Residues of sugar derivatives such as sialic acid; maltose, maltotriose, maltotetraose, maltopentaose, maltohexaose, maltoheptaose, lactose, gentiobiose, xylobiose, primebellose, melibiose, lutinose, vicyanose, laminaribiose Residues of oligosaccharides composed of 2 to 7 sugars such as cellobiose, cellotriose and panose, and the like, and particularly preferred are residues of oligosaccharides composed of glucose and glucose. The hydroxyl groups at the 2, 3, 4, and 6 positions may be modified with, for example, an acetyl group, a malonyl group, a phosphate group, or a methyl group.

上記構造式１にて表される化合物の内、下記構造式２にて表されるウィタノシドＩＶが、その効果が高く、特に好ましく用いることができる。   Of the compounds represented by Structural Formula 1, Vitanoside IV represented by Structural Formula 2 below is highly effective and can be particularly preferably used.

構造式２
Structural formula 2

本発明の脊髄損傷治療剤として用いられるウィタノシド（Ｗｉｔｈａｎｏｓｉｄｅ）ＩＶ及びその周辺化合物は、アシュワガンダ（Ａｓｈｗａｇａｎｄｈａ：Ｗｉｔｈａｎｉａ ｓｏｍｎｉｆｅｒａ Ｄｕｎａｌの根）から単離することができる。例えば、Ｃｈｅｍ．Ｐｈａｒ．Ｂｕｌｌ．，第５０巻，ｐ７６０−７６５（２００２年）に記載の方法により単離することが可能である。また、化学合成手法により合成したものでも用いることができ、その方法は特に限定されない。   Withanoside IV and its peripheral compounds used as a therapeutic agent for spinal cord injury of the present invention can be isolated from Ashwagandha (root of Whitania soniifera Dunal). For example, Chem. Phar. Bull. 50, p760-765 (2002). Moreover, what was synthesize | combined with the chemical synthesis method can also be used, and the method is not specifically limited.

また、ウィタノシドの代謝物として得ることも出来る。たとえば構造式１で包含される化合物の１つであるソミノン（ｓｏｍｉｎｏｎｅ、Ｒ1＝Ｈ、Ｒ2＝ＯＨ、Ｒ3＝ＯＨ）はウィタノシドＩＶの体内における代謝物であり、例えばＨｅｔｅｒｏｃｙｃｌｅｓ，第３４巻，ｐ６８９−６９８（１９９２年）に記載の方法で得ることができる。具体的には、ウィタノシドＩＶを酵素、例えばナリンギナーゼ（ｎａｒｉｎｇｉｎａｓｅ）で、ｐＨ５．２５、３７℃の条件で３日間処理し、分配高速液体クロマトグラフィーなどにより分離、精製することで得ることができる。 It can also be obtained as a metabolite of withanoside. For example, one of the compounds encompassed by structural formula 1 (somone, R 1 = H, R 2 = OH, R 3 = OH) is a metabolite of witanoside IV, for example, Heterocycles, Vol. 34. , P689-698 (1992). Specifically, it can be obtained by treating Witanoside IV with an enzyme such as naringinase at pH 5.25 and 37 ° C. for 3 days, and separating and purifying it by partition high performance liquid chromatography.

本発明の食品組成物は、特に脊髄損傷患者用食品に適用することができる。本発明の薬剤を含有する特定保健用食品等の特別用途食品や栄養機能食品として直接摂取することにより脊髄損傷回復治療を簡便に行うことができる。   The food composition of the present invention can be applied particularly to food for patients with spinal cord injury. Spinal cord injury recovery treatment can be easily performed by ingesting directly as a special-purpose food such as a food for specified health use or a food with a nutritional function containing the drug of the present invention.

具体的には、食品組成物として使用する場合には、各種飲食品（牛乳、清涼飲料、発酵乳、ヨーグルト、チーズ、パン、ビスケット、クラッカー、ピッツァクラスト、調製粉乳、流動食、病者用食品、幼児用粉乳等食品、授乳婦用粉乳等食品、栄養食品等）に本発明の薬剤を添加し、これを摂取してもよい。本有効成分をそのまま使用したり、他の食品ないし食品成分と混合するなど、通常の食品組成物における常法にしたがって使用できる。また、その性状についても、通常用いられる飲食品の状態、例えば、固体状（粉末、顆粒状その他）、ペースト状、液状ないし懸濁状のいずれでもよい。   Specifically, when used as a food composition, various foods and drinks (milk, soft drinks, fermented milk, yogurt, cheese, bread, biscuits, crackers, pizza crusts, prepared milk powder, liquid foods, food for the sick The drug of the present invention may be added to foods such as infant milk powder, foods such as lactating milk powder, and nutritional foods. The present active ingredient can be used as it is, or can be used according to a conventional method for ordinary food compositions such as mixing with other foods or food ingredients. Moreover, about the property, the state of the food / beverage products normally used, for example, any of solid (powder, granule, etc.), paste, liquid or suspension may be sufficient.

その他の成分についても特に限定されないが、本発明の薬剤を含有する飲食物には、水、タンパク質、糖質、脂質、ビタミン類、ミネラル類、有機酸、有機塩基、果汁、フレーバー類等を主成分として使用することができる。タンパク質としては、例えば全脂粉乳、脱脂粉乳、部分脱脂粉乳、カゼイン、ホエイ粉、ホエイタンパク質、ホエイタンパク質濃縮物、ホエイタンパク質分離物、α―カゼイン、β―カゼイン、κ−カゼイン、β―ラクトグロブリン、α―ラクトアルブミン、ラクトフェリン、大豆タンパク質、鶏卵タンパク質、肉タンパク質等の動植物性タンパク質、これら加水分解物；バター、乳性ミネラル、クリーム、ホエイ、非タンパク態窒素、シアル酸、リン脂質、乳糖等の各種乳由来成分などが挙げられる。糖類、加工澱粉（テキストリンのほか、可溶性澱粉、ブリティッシュスターチ、酸化澱粉、澱粉エステル、澱粉エーテル等）、食物繊維などが挙げられる。脂質としては、例えば、ラード、魚油等、これらの分別油、水素添加油、エステル交換油等の動物性油脂；パーム油、サフラワー油、コーン油、ナタネ油、ヤシ油、これらの分別油、水素添加油、エステル交換油等の植物性油脂などが挙げられる。ビタミン類としては、例えば、ビタミンＡ、カロチン類、ビタミンＢ群、ビタミンＣ、ビタミンＤ群、ビタミンＥ、ビタミンＫ群、ビタミンＰ、ビタミンＱ、ナイアシン、ニコチン酸、パントテン酸、ビオチン、イノシトール、コリン、葉酸などが挙げられ、ミネラル類としては、例えば、カルシウム、カリウム、マグネシウム、ナトリウム、銅、鉄、マンガン、亜鉛、セレンなどが挙げられる。有機酸としては、例えば、リンゴ酸、クエン酸、乳酸、酒石酸などが挙げられる。これらの成分は、２種以上を組み合わせて使用することができ、合成品及び／又はこれらを多く含む食品を用いてもよい。   Other ingredients are not particularly limited, but foods and drinks containing the drug of the present invention mainly include water, proteins, carbohydrates, lipids, vitamins, minerals, organic acids, organic bases, fruit juices, flavors and the like. Can be used as an ingredient. Examples of proteins include whole milk powder, skim milk powder, partially skim milk powder, casein, whey powder, whey protein, whey protein concentrate, whey protein isolate, α-casein, β-casein, κ-casein, β-lactoglobulin , Α-lactalbumin, lactoferrin, soy protein, chicken egg protein, meat protein and other animal and plant proteins, hydrolysates thereof; butter, milk minerals, cream, whey, non-protein nitrogen, sialic acid, phospholipid, lactose, etc. And various milk-derived components. Examples include saccharides, processed starch (in addition to text phosphorus, soluble starch, British starch, oxidized starch, starch ester, starch ether, etc.), dietary fiber, and the like. Examples of the lipid include animal oils such as lard, fish oil, etc., fractionated oils, hydrogenated oil, transesterified oil, etc .; palm oil, safflower oil, corn oil, rapeseed oil, coconut oil, fractionated oils thereof, Examples include vegetable oils such as hydrogenated oils and transesterified oils. Examples of vitamins include vitamin A, carotene, vitamin B group, vitamin C, vitamin D group, vitamin E, vitamin K group, vitamin P, vitamin Q, niacin, nicotinic acid, pantothenic acid, biotin, inositol, choline. Examples of minerals include calcium, potassium, magnesium, sodium, copper, iron, manganese, zinc, and selenium. Examples of the organic acid include malic acid, citric acid, lactic acid, and tartaric acid. These components can be used in combination of two or more, and synthetic products and / or foods containing a large amount thereof may be used.

本発明の脊髄損傷治療剤これを有効成分とする食品・医薬組成物の投与量は、投与経路、ヒトを含む投与対象動物の年齢、体重、症状など、種々の要因を考慮して、適宜設定することができる。本発明はこれに限定されないが、好ましくは、有効成分として０．０１〜１００μｍｏｌ／ｋｇ／ｄａｙが適当である。
また本発明の脊髄損傷治療剤、医薬組成物は、経口投与又は非経口投与（筋肉内、皮下、静脈内、坐薬、経皮等）のいずれでも投与できる。 The therapeutic agent for spinal cord injury of the present invention The dosage of the food / pharmaceutical composition comprising this as an active ingredient is appropriately set in consideration of various factors such as the route of administration and the age, weight and symptoms of animals to be administered including humans. can do. Although this invention is not limited to this, Preferably 0.01-100 micromol / kg / day is suitable as an active ingredient.
The therapeutic agent for spinal cord injury and the pharmaceutical composition of the present invention can be administered either orally or parenterally (intramuscular, subcutaneous, intravenous, suppository, transdermal, etc.).

本発明による脊髄損傷治療剤、医薬組成物の投与形態としては、例えば錠剤、被覆錠剤、カプセル剤、顆粒剤、散剤、溶液、シロップ剤、乳液等による経口投与をあげることができる。これらの各種製剤は、常法に従って主薬である本発明の脊髄損傷治療剤に賦形剤、結合剤、崩壊剤、滑沢剤、着色剤、矯味矯臭剤、溶解補助剤、懸濁剤、コーティング剤などの医薬の製剤技術分野において通常使用しうる既知の補助剤を用いて製剤化することができる。   Examples of the administration form of the spinal cord injury therapeutic agent and pharmaceutical composition according to the present invention include oral administration using tablets, coated tablets, capsules, granules, powders, solutions, syrups, emulsions and the like. These various preparations are prepared according to conventional methods, including the excipients, binders, disintegrants, lubricants, coloring agents, flavoring agents, solubilizers, suspension agents, and coatings. It can be formulated using known adjuvants that can be generally used in the pharmaceutical formulation technical field such as pharmaceuticals.

本発明の脊髄損傷治療剤の量は、その目的、用途（予防剤、治療剤等の医薬品組成物）に応じて任意に定めることができ。本発明はこれに限定されないがその含量としては、全体量に対して通常、０．００１〜１００％（ｗ／ｗ）、特に０．１〜１００％が好ましい。   The amount of the spinal cord injury therapeutic agent of the present invention can be arbitrarily determined according to the purpose and application (pharmaceutical composition such as prophylactic agent and therapeutic agent). Although this invention is not limited to this, As the content, 0.001 to 100% (w / w) normally with respect to the whole quantity, Especially 0.1 to 100% is preferable.

以下、本発明について実施例を挙げて説明するが、本発明はこれらにより限定されるものではない。   EXAMPLES Hereinafter, although an Example is given and this invention is demonstrated, this invention is not limited by these.

［脊髄損傷モデルマウスの作製］
マウス（ｄｄＹ、雄、６週齢）の背部皮膚および筋肉をエーテル麻酔下で切開し、脊椎を露出させた。次いで、胸椎第８−１１番レベルの椎弓を電気ドリルで切除した。露出した胸椎に、錘（２．５ｇ）を２ｃｍ垂直上方から３回落下させ、脊髄を損傷させた。
一方、椎弓切除まで施術したものをＳｈａｍ群とした。
この時、脊髄損傷モデルマウスは、両方の後肢が麻痺しており、後肢で体重を支えて動くことは出来ない状態であった。また、Ｓｈａｍ群は正常な運動機能が保持されていることを確認した。 [Production of spinal cord injury model mice]
The dorsal skin and muscle of a mouse (ddY, male, 6 weeks old) were dissected under ether anesthesia to expose the spine. Next, the vertebral arch at level 8-11 of the thoracic vertebra was excised with an electric drill. A weight (2.5 g) was dropped 3 times from 2 cm vertically above the exposed thoracic vertebra, and the spinal cord was damaged.
On the other hand, the sham group was treated to the laminectomy.
At this time, in the spinal cord injury model mouse, both hind limbs were paralyzed, and the hind limbs were unable to support their weight and move. In addition, it was confirmed that the Sham group maintained normal motor function.

［薬物投与］
脊髄を損傷させた後１時間経過後に、第１回の薬物投与を行った。次いで、翌日から１日１回、２０日間の連続経口投与を行った。ウィタノシドＩＶは蒸留水に溶解し、１０ｍｏｌ／ｋｇ／ｄａｙの用量で投与した。なお、Ｓｈａｍ群及び対照群にはビヒクルとして水を経口投与した。なお、ここで使用したウィタノシドＩＶはアシュワガンダからのメタノール抽出後の単離品であり、ＨＰＬＣによりその純度を確認した（検出：ＵＶ２２０ｎｍ）。 [Drug administration]
The first drug administration was performed 1 hour after the spinal cord was damaged. Subsequently, continuous oral administration for 20 days was performed once a day from the next day. Withanoside IV was dissolved in distilled water and administered at a dose of 10 mol / kg / day. In addition, water was orally administered as a vehicle to the Sham group and the control group. The witanoside IV used here is an isolated product after methanol extraction from Ashwagandha, and its purity was confirmed by HPLC (detection: UV 220 nm).

［行動学的評価］
以下の行動学的評価を脊髄損傷の翌日より２０日目まで行った。
１．Ｂａｓｓｏ，Ｂｅａｔｔｉｅ，ａｎｄ Ｂｒｅｓｎａｈａｎ（ＢＢＢ）ｓｃａｌｅ
現在脊髄損傷動物の評価では最も一般的なスコアリング手法であるＢＢＢｓｃａｌｅ（バッソ等（Ｂａｓｓｏ，Ｂｅａｔｔｉｅ，Ｂｒｅｓｎａｈａｎ），Ｅｘｐｅｒｉｍｅｎｔａｌ Ｎｅｕｒｏｌｏｇｙ，１９９６年，第１３９巻，ｐ．２４４−２５６）によって評価した。すなわち、ケージ内でのマウスの後肢機能を５分間目視にて観察し、ＢＢＢｓｃａｌｅに基づいて０〜２１点にスコアリングした。
２．立ち上がり行動
ケージ内での後肢による立ち上がり回数を５分間計測した。 [Behavioral evaluation]
The following behavioral evaluation was performed from the day after the spinal cord injury to the 20th day.
1. Basso, Beattie, and Bresnahan (BBB) scale
BBBscale (Basso, Beattie, Bresnahan, Experimental Neurology, 1996, Vol. 139, pp. 244-256), which is currently the most common scoring technique, is used to evaluate animals with spinal cord injury. That is, the hind limb function of the mouse in the cage was visually observed for 5 minutes and scored from 0 to 21 points based on the BBBscale.
2. Standing behavior The number of standing by the hind limbs in the cage was measured for 5 minutes.

［免疫組織染色］
脊髄損傷処理後の２１日目のマウスを麻酔し、左心室から生理食塩水を注入し脱血し、続けて４％パラホルムアルデヒド−ＰＢＳ（ＰＦＡ）を注入し還流固定した。脊髄を脊椎ごと摘出し４％ＰＦＡ中で一晩固定した。脊椎を開け、脊髄を摘出し、損傷部位を中心に２ｃｍ長の胸椎部をカットした。１０％、２０％、３０％スクロース溶液で順次置換した後、ＯＣＴ ｃｏｍｐｏｕｎｄにて包埋し、クリオスタットにより１２μｍ厚のｓａｇｉｔａｌ ｓｌｉｃｅを作製し、ゼラチンコーティングしたスライドグラスに貼り付けた。 [Immunohistochemical staining]
The mouse on the 21st day after the spinal cord injury treatment was anesthetized, and saline was injected from the left ventricle to remove blood, followed by infusion of 4% paraformaldehyde-PBS (PFA) and fixation under reflux. The spinal cord was removed and fixed overnight in 4% PFA. The spine was opened, the spinal cord was removed, and a 2 cm long thoracic vertebra was cut around the damaged site. Subsequent replacement with 10%, 20%, and 30% sucrose solutions was followed by embedding with an OCT compound, and a 12 μm thick sagittal slice was prepared with a cryostat and attached to a gelatin-coated slide glass.

ｓｌｉｃｅの周囲をＤＡＫＯ ＰＥＮ（ダコ・ジャパン株式会社製）で囲み、４％ＰＦＡを滴下し３０分間、固定を行った。一次抗体反応には、マウス抗リン酸化型ｎｅｕｒｏｆｉｌａｍｅｎｔ−Ｈ（ＮＦ−Ｈ）モノクローナル抗体（希釈倍率 １：１０００）（Ｃｌｏｎｅ ＳＭＩ３１２，Ｓｔｅｒｎｂｅｒｇｅｒ Ｍｏｎｏｃｌｏｎａｌｓ，Ｌｕｔｈｅｒｖｉｌｅ，メリーランド州、米国）、ウサギ抗ｍｙｅｌｉｎ ｂａｓｉｃ ｐｒｏｔｅｉｎ（ＭＢＰ）ポリクローナル抗体（希釈倍率 １：１００）（Ｃｈｅｍｉｃｏｎ，Ｔｅｍｅｃｕｌａ，カリフォルニア州、米国）、ウサギ抗ｐｅｒｉｐｈｅｒａｌ ｍｙｅｌｉｎ ｐｒｏｔｅｉｎ（ＰＭＰ）−２２ポリクローナル抗体（希釈倍率 １：５００）（Ｌａｂ Ｖｉｓｉｏｎ，Ｆｒｅｍｏｎｔ，カリフォルニア州、米国）をそれぞれ用いた。４℃で一晩反応後、２次抗体にはＡｌｅｘａ Ｆｌｕｏｒ ４８８標識ヤギ抗マウスＩｇＧ抗体と、Ａｌｅｘａ Ｆｌｕｏｒ ５６８標識ヤギ抗ウサギＩｇＧ抗体（いずれも希釈倍率 １：２００）（Ｍｏｌｅｃｕｌａｒ Ｐｒｏｂｅｓ，Ｃａｒｌｓｂａｄ，カリフォルニア州、米国）とを用いた。二重蛍光免疫染色された試料の灰白質、白質の損傷部位とその前後を蛍光顕微鏡ＡＸ−８０（オリンパス株式会社製）を用いて拡大観察し、またその蛍光画像を取得し、抗体に陽性の領域の積算値をＡＴＴＯ Ｄｅｎｓｉｔｏｇｒａｐｈ（アトー株式会社製）によって測定した。   The periphery of the slice was surrounded by DAKO PEN (manufactured by Dako Japan Co., Ltd.), and 4% PFA was dropped and fixed for 30 minutes. For the primary antibody reaction, mouse anti-phosphorylated neurofilament-H (NF-H) monoclonal antibody (dilution ratio 1: 1000) (Clone SMI312, Sternberger Monoclonals, Lutherville, Md., USA), rabbit anti-myelin basic protein ( MBP) polyclonal antibody (dilution ratio 1: 100) (Chemicon, Temecula, CA, USA), rabbit anti-peripheral myelin protein (PMP) -22 polyclonal antibody (dilution ratio 1: 500) (Lab Vision, Fremont, CA) US) was used. After overnight reaction at 4 ° C., the secondary antibodies were Alexa Fluor 488-labeled goat anti-mouse IgG antibody and Alexa Fluor 568-labeled goat anti-rabbit IgG antibody (both dilution ratio 1: 200) (Molecular Probes, Carlsbad, CA) , USA). Double-fluorescence immunostained sample gray matter, white matter damage site and its front and back are magnified using a fluorescence microscope AX-80 (manufactured by Olympus Co., Ltd.), and the fluorescence image is acquired and positive for the antibody. The integrated value of the area was measured by ATTO Densitograph (manufactured by ATTO Corporation).

［データ解析］
データは平均値±標準誤差（ｓ．ｅ．ｍ．）で表した。有意差検定は分散分析（ＡＮＯＶＡ）、あるいはｔｗｏ ｗａｙ ｒｅｐｅａｔｅｄ ｍｅａｓｕｒｅｄ ＡＮＯＶＡにより行い、ｐｏｓｔ ｈｏｃ ｔｅｓｔ はＤｕｎｎｅｔｔ’ｓ ｔｅｓｔにより行った。有意水準は５％とした。 [Data analysis]
Data are expressed as mean ± standard error (sem). Significance test was performed by analysis of variance (ANOVA) or two way repeated measured ANOVA, and post hoc test was performed by Dunnett's test. The significance level was 5%.

［実験結果］
以下、得られた結果を図示のグラフ並びに組織の顕微鏡写真を参照しながら説明する。
なお、各群あたりの個体数は４〜６匹とした。
脊髄損傷モデルマウス（対照群）の後肢機能障害は、図１に示すＢＢＢ ｓｃａｌｅによっても、図２に示す後肢による立ち上がり回数によっても、Ｓｈａｍ群と比べて有意に低下していることが分かる。一方、脊髄損傷モデルマウスのウィタノシドＩＶ経口投与群では、後肢機能障害が、対照群と比べてＢＢＢ ｓｃａｌｅによる判定においても、後肢による立ち上がり回数においても有意に改善されていることが分かった。 [Experimental result]
Hereinafter, the obtained results will be described with reference to the illustrated graph and the micrograph of the tissue.
The number of individuals per group was 4-6.
It can be seen that the hindlimb dysfunction of the spinal cord injury model mouse (control group) is significantly lower than that of the Sham group, both by the BBB scale shown in FIG. 1 and by the number of rises by the hindlimb shown in FIG. On the other hand, in the Witanoside IV oral administration group of spinal cord injury model mice, it was found that hind limb dysfunction was significantly improved both in the determination by BBB scale and in the number of rises by the hind limb as compared with the control group.

また、図３に示す軸索組織については、Ｓｈａｍ群では水平方向に伸展（ＮＦ−Ｈ陽性）していることが分かる（図３Ａ、図３Ｇ参照）。また、中枢性ミエリンの発現（ＭＢＰ陽性）が認められた（図３Ｄ、図３Ｇ参照）。
これに対して、対照群では軸索組織が断裂し、特に損傷中心部では断裂した軸索線維が固まっていることが分かる（図３Ｂ、図３Ｈ参照）。また、対照群では中枢性ミエリンの脱落が見られた（図３Ｅ、図３Ｈ参照）。
一方、本発明の脊髄損傷治療剤を投与したウィタノシドＩＶ経口投与群では、損傷中心部に軸索線維の塊は見られず、代わりにＳｈａｍ群のような水平方向の軸索伸展が観察された（図３Ｃ）。一方、中枢性ミエリンは脱落したままであった（図３Ｆ、図３Ｉ参照）。 Moreover, about the axon tissue shown in FIG. 3, it turns out that it is extended in the horizontal direction (NF-H positive) in the Sham group (refer FIG. 3A and FIG. 3G). Moreover, the expression of central myelin (MBP positive) was observed (see FIGS. 3D and 3G).
On the other hand, in the control group, it can be seen that the axonal tissue is torn, and in particular, the torn axon fibers are solidified at the damage center (see FIGS. 3B and 3H). In the control group, central myelin was eliminated (see FIGS. 3E and 3H).
On the other hand, in the oral administration group of Withanoside IV administered with the therapeutic agent for spinal cord injury of the present invention, no axonal fiber mass was observed in the center of the injury, and instead horizontal axon extension was observed as in the Sham group. (FIG. 3C). On the other hand, central myelin remained missing (see FIGS. 3F and 3I).

末梢性ミエリン検出を試みたところ、対照群では末梢性ミエリンがわずかに増加していた（図８Ｅ、図８Ｈ参照）。これに対して、本発明のウィタノシドＩＶ経口投与群では損傷部位、特に灰白質の軸索の周囲に末梢性ミエリンが増加していることが分かった（図８Ｆ、図８Ｉ参照）。   Attempts were made to detect peripheral myelin, and there was a slight increase in peripheral myelin in the control group (see FIGS. 8E and 8H). On the other hand, it was found that peripheral myelin was increased in the woundanoside IV oral administration group of the present invention, particularly around the axon of gray matter (see FIGS. 8F and 8I).

以上の結果から、明らかに対照群に比べて本発明のウィタノシドＩＶ経口投与群では損傷部位の修復作用が見られた。   From the above results, it was clear that the repaired effect of the damaged site was observed in the oral administration group of the Witanoside IV of the present invention as compared with the control group.

上記したように本発明によるウィタノシド（Ｗｉｔｈａｎｏｓｉｄｅ）ＩＶ及びその周辺化合物により、簡便かつ高い効果を示す脊髄損傷治療剤又は食品・医薬品組成物を提供することが可能となった。
また、上記薬剤又は食品・医薬品組成物により、簡便にヒトなどの脊髄治療目的とした効果的な治療方法を行うことができる。 As described above, it is possible to provide a therapeutic agent for spinal cord injury or a food / pharmaceutical composition that shows a simple and high effect by using the witanoside IV and its peripheral compounds according to the present invention.
Moreover, an effective treatment method for the purpose of treating spinal cords of humans or the like can be easily performed with the above-mentioned drug or food / pharmaceutical composition.

マウスのＢＢＢｓｃａｌｅの判定を示すスコアグラフ図（脊髄損傷後２０日目まで）である。○はＳｈａｍ群を、●は対照群を、□は本発明によるウィタノシドＩＶ投与群を示す。It is a score graph figure (until the 20th day after spinal cord injury) which shows determination of BBBscale of a mouse | mouth. ◯ indicates the Sham group, ● indicates the control group, and □ indicates the Witanoside IV administration group according to the present invention. マウスのケージ内での後肢による立ち上がり回数のグラフ図（脊髄損傷後２０日目まで）である。○はＳｈａｍ群を、●は対照群を、□は本発明によるウィタノシドＩＶ投与群を示す。It is a graph (up to the 20th day after spinal cord injury) of the number of rising by the hind limb in the cage of the mouse. ◯ indicates the Sham group, ● indicates the control group, and □ indicates the Witanoside IV administration group according to the present invention. 脊髄損傷後２１日目のマウスの胸椎切片を抗リン酸化ＮＦ−Ｈモノクローナル抗体（Ａ，Ｂ，Ｃ）と抗ＭＢＰポリクローナル抗体（Ｄ，Ｅ，Ｆ）を用いて二重蛍光免疫染色した組織の顕微鏡写真である。左側が頭側、右側が尾側、矢印は圧挫部を示す。「Ｓｈａｍ」はＳｈａｍ群を、「Ｃｏｎｔ」は対照群を、「ＷＳ−ＩＶ」は本発明によるウィタノシドＩＶ投与群を示す（以下の図面において同様記述は同様の意味を示す）。なお、Ｇ，Ｈ，Ｉは、それぞれＡにＤを、ＢにＥを、またＣにＦを重ね合わせたもので観察倍率をあげた写真である。A tissue of a thoracic spine section of a mouse 21 days after spinal cord injury was subjected to double fluorescent immunostaining using anti-phosphorylated NF-H monoclonal antibody (A, B, C) and anti-MBP polyclonal antibody (D, E, F). It is a micrograph. The left side is the head side, the right side is the caudal side, and the arrow indicates the crushing part. “Sham” indicates the Sham group, “Cont” indicates the control group, and “WS-IV” indicates the Witanoside IV administration group according to the present invention (similar descriptions in the following drawings have the same meaning). G, H, and I are photographs in which the magnification of observation is increased by superposing D on A, E on B, and F on C, respectively. 抗リン酸化ＮＦ−Ｈモノクローナル抗体に対する灰白質の陽性領域の蛍光強度を示すグラフ図である。Ｒｏｓｔｒａｌは頭側、Ｃａｕｄａｌは尾側を示す。It is a graph which shows the fluorescence intensity of the positive region of the gray matter with respect to an anti- phosphorylated NF-H monoclonal antibody. “Rostral” indicates the head side and “Caudal” indicates the caudal side. 抗リン酸化ＮＦ−Ｈモノクローナル抗体に対する白質の陽性領域の蛍光強度を示すグラフ図である。Ｒｏｓｔｒａｌは頭側、Ｃａｕｄａｌは尾側を示す。It is a graph which shows the fluorescence intensity of the positive region of the white matter with respect to an anti- phosphorylated NF-H monoclonal antibody. “Rostral” indicates the head side and “Caudal” indicates the caudal side. 抗ＭＢＰポリクローナル抗体に対する灰白質及び白質の陽性領域の蛍光強度を示すグラフ図である。It is a graph which shows the fluorescence intensity of the gray matter and white matter positive area | region with respect to an anti-MBP polyclonal antibody. 脊髄損傷後２１日目のマウスの胸椎切片を抗リン酸化ＮＦ−Ｈモノクローナル抗体（左）と抗ＭＢＰポリクローナル抗体（右）を用いて二重蛍光免疫染色した組織の顕微鏡写真である。Ｗは白質部を、Ｇは灰白質部を示す。It is the microscope picture of the structure | tissue which carried out the double fluorescence immunostaining of the thoracic-vertebral section of the mouse | mouth 21st after spinal cord injury using the anti-phosphorylated NF-H monoclonal antibody (left) and the anti-MBP polyclonal antibody (right). W represents a white matter portion and G represents a gray matter portion. 脊髄損傷後２１日目のマウスの胸椎切片を抗リン酸化ＮＦ−Ｈモノクローナル抗体（Ａ，Ｂ，Ｃ）と抗ＰＭＰ−２２ポリクローナル抗体（Ｄ，Ｅ，Ｆ）を用いて二重蛍光免疫染色した組織の顕微鏡写真である。左側が頭側、右側が尾側、矢印は圧挫部を示す。ＳｈａｍはＳｈａｍ群を、Ｃｏｎｔは対照群を、ＷＳ−ＩＶは本発明によるウィタノシドＩＶ投与群を示す。なお、Ｇ，Ｈ，Ｉは、それぞれＡにＤを、ＢにＥを、またＣにＦを重ね合わせたもので観察倍率をあげた写真である。Thoracic vertebral sections of mice 21 days after spinal cord injury were double fluorescently immunostained using anti-phosphorylated NF-H monoclonal antibodies (A, B, C) and anti-PMP-22 polyclonal antibodies (D, E, F). It is a microscope picture of a structure | tissue. The left side is the head side, the right side is the caudal side, and the arrow indicates the crushing part. Sham represents a Sham group, Cont represents a control group, and WS-IV represents a Witanoside IV administration group according to the present invention. G, H, and I are photographs in which the magnification of observation is increased by superposing D on A, E on B, and F on C, respectively. 抗ＰＭＰ−２２ポリクローナル抗体に対する灰白質及び白質の陽性領域の蛍光強度を示すグラフ図である。It is a graph which shows the fluorescence intensity of the gray matter and white matter positive area | region with respect to an anti- PMP-22 polyclonal antibody. 脊髄損傷後２１日目のマウスの胸椎切片をＰＭＰ−２２ポリクローナル抗体を用いて蛍光免疫染色した組織の顕微鏡写真である。Ｗは白質部を、Ｇは灰白質部を示す。It is a microscope picture of the structure | tissue which carried out the fluorescence immunostaining of the thoracic spine section of the mouse | mouth 21st after spinal cord injury using the PMP-22 polyclonal antibody. W represents a white matter portion and G represents a gray matter portion.

Claims (4)

下記構造式１の化合物の少なくとも１種を有効成分とする脊髄損傷治療剤。
構造式１
（式中、Ｒ1及びＲ2はそれぞれ独立して、水素原子、水酸基、アルキル基、またはアルコキシ基であり、Ｒ3は水素原子、水酸基、アルキル基、アルコキシ基、または糖残基である。） A therapeutic agent for spinal cord injury comprising at least one compound of the following structural formula 1 as an active ingredient.
Structural formula 1
(In the formula, R 1 and R 2 are each independently a hydrogen atom, a hydroxyl group, an alkyl group, or an alkoxy group, and R 3 is a hydrogen atom, a hydroxyl group, an alkyl group, an alkoxy group, or a sugar residue. ) 下記構造式２のウィタノシドＩＶを有効成分とする脊髄損傷治療剤。
構造式２
A therapeutic agent for spinal cord injury comprising withanoside IV of the following structural formula 2 as an active ingredient.
Structural formula 2
請求項１又は２に記載の脊髄損傷治療剤を有効成分として含有し、脊髄損傷の回復作用を有する食品・医薬品組成物。   A food / pharmaceutical composition comprising the therapeutic agent for spinal cord injury according to claim 1 or 2 as an active ingredient and having an action of recovering spinal cord injury. 請求項１若しくは２に記載の脊髄損傷治療剤又は請求項３に記載の食品・医薬品組成物の有効量を投与することを特徴とする脊髄損傷の回復方法。
A method for recovering spinal cord injury, comprising administering an effective amount of the therapeutic agent for spinal cord injury according to claim 1 or 2 or the food / pharmaceutical composition according to claim 3.
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