WO2020004463A1 - Composition for preventing or treating diabetic peripheral neuropathy - Google Patents

Composition for preventing or treating diabetic peripheral neuropathy Download PDF

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Publication number
WO2020004463A1
WO2020004463A1 PCT/JP2019/025391 JP2019025391W WO2020004463A1 WO 2020004463 A1 WO2020004463 A1 WO 2020004463A1 JP 2019025391 W JP2019025391 W JP 2019025391W WO 2020004463 A1 WO2020004463 A1 WO 2020004463A1
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peripheral neuropathy
preventing
diabetic peripheral
composition
treating diabetic
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PCT/JP2019/025391
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French (fr)
Japanese (ja)
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升三 西田
正寛 椿
朋也 武田
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学校法人近畿大学
小林製薬株式会社
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Publication of WO2020004463A1 publication Critical patent/WO2020004463A1/en

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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
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/06—Fungi, e.g. yeasts
    • A61K36/07—Basidiomycota, e.g. Cryptococcus
    • A—HUMAN NECESSITIES
    • 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
    • A61P25/02—Drugs for disorders of the nervous system for peripheral neuropathies
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor

Definitions

  • the present invention relates to a composition for preventing or treating diabetes-induced peripheral neuropathy (hereinafter, sometimes referred to as diabetic peripheral neuropathy).
  • diabetes-induced peripheral neuropathy hereinafter, sometimes referred to as diabetic peripheral neuropathy.
  • Diabetic peripheral neuropathy is a complication of diabetes. It is known that peripheral neuropathy such as hyperesthesia or hypoesthesia occurs as diabetes progresses. This peripheral neuropathy is a major problem because the quality of daily life of patients is reduced. However, at present, no effective treatment has been established for diabetic peripheral neuropathy. There is a need for a method for preventing and / or treating diabetic peripheral neuropathy.
  • Patent Document 1 discloses that a lactam compound is effective as a sugar transport-enhancing agent, preventing diabetes, diabetic peripheral neuropathy, diabetic nephropathy, diabetic macroangiopathy, impaired glucose tolerance, or preventing obesity and And / or can be used as a therapeutic agent.
  • Patent Document 1 discloses that a lactam compound has a sugar transporting ability, including Examples, but does not specifically describe peripheral neuropathy. Patent Document 1 is limited to the disclosure that it has an effect of lowering the blood sugar level.
  • tricyclic antidepressants pregabalin as an ⁇ 2 ⁇ ligand, duloxetine as a serotonin / noradrenaline reuptake inhibitor, and the like have been recommended as treatments for peripheral neuropathy, but the side effects cannot be ignored. In addition, these drugs cannot be used as a composition for preventing diabetic peripheral neuropathy.
  • An object of the present invention is to provide a composition for preventing or treating diabetic peripheral neuropathy, which is capable of preventing and treating diabetic peripheral neuropathy.
  • the present inventors searched for a material capable of preventing or treating diabetic peripheral neuropathy, and found a shiitake mycelium.
  • the present inventors have clarified that the shiitake mushroom mycelium exhibits an excellent preventive or therapeutic effect on diabetic peripheral neuropathy, and completed the present invention.
  • the first invention is a composition for preventing or treating diabetic peripheral neuropathy, comprising an extract of Shiitake mushroom mycelium.
  • the second invention is a pharmaceutical composition for preventing or treating diabetic peripheral neuropathy, comprising an extract of Shiitake mushroom mycelium.
  • the second invention is a pharmaceutical composition for preventing or treating diabetic peripheral neuropathy, wherein the daily dose of the extract of Shiitake mushroom mycelium is 4 mg / kg body weight or more as a dry powder weight. .
  • the third invention is a processed food having an effect of preventing or treating diabetic peripheral neuropathy, comprising an extract of Shiitake mushroom mycelium.
  • a fourth invention provides a method for preventing or treating diabetic peripheral neuropathy, comprising administering to a subject the pharmaceutical composition for preventing or treating diabetic peripheral neuropathy according to the second invention. Excluding methods of surgery, treatment, and diagnosis).
  • the present invention may also be referred to as prevention or treatment of diabetic peripheral neuropathy using a Shiitake mushroom mycelium extract.
  • the present invention can prevent or treat diabetic peripheral neuropathy. That is, by administering the Shiitake mushroom mycelium extract, numbness of the limbs induced by diabetes, pain in the limbs, decreased deep tendon reflex, decreased muscular strength, allodynia, hyperalgesia, elaborate dysfunction of the fingers, walking disorders, The stumbling, falling, bending disorder (difficult or impossible such as sitting straight, cross-legged, sitting sideways or sitting on a chair), or paralysis of limbs are improved.
  • the composition for preventing or treating diabetic peripheral neuropathy according to the present invention can exert its effects when taken orally. Therefore, there is no need to go to the hospital and the burden on the patient is small.
  • the improvement of diabetic peripheral neuropathy also improves the quality of life of the patient.
  • composition for preventing or treating diabetic peripheral neuropathy according to the present invention exerts a preventive effect on diabetic peripheral neuropathy, so that when taken together with daily food, it can also be used to prevent diabetic peripheral neuropathy. effective.
  • composition for preventing or treating diabetic peripheral neuropathy according to the present invention will be described with reference to the drawings and examples. Note that the following description exemplifies one embodiment and one example of the present invention, and the present invention is not limited to the following description. The following description can be modified without departing from the spirit of the present invention.
  • prevention includes the concepts of prevention, early treatment, and prevention of functional decline.
  • treatment is not necessarily limited to the improvement of the symptoms of the affected patient, but is a concept that also includes the alleviation and improvement of the symptoms of a healthy person (preliminary group).
  • “Lentinus edodes mycelium”, which is a raw material of the shiitake mushroom mycelium extract used in the composition for preventing or treating diabetic peripheral neuropathy according to the present invention, is not particularly limited as long as it is a shiitake mushroom mycelium. Not done.
  • a mycelium in a stage before the edible fruiting body can be used.
  • an extract (Lentinus edodes mycelium extract) obtained by culturing Lentinus edodes on a solid medium can be used.
  • the Shiitake mushroom mycelium extract used in the composition for preventing or treating diabetic peripheral neuropathy according to the present invention can be prepared by a method known in the art.
  • an extract obtained by extracting a shiitake mycelium after pulverization can be suitably used as a shiitake mushroom mycelium extract.
  • an extract obtained by pulverizing and decomposing a solid medium containing Shiitake mycelium in the presence of water can also be used.
  • Examples of the solvent used for the preparation of the extract include water, ethanol, methanol, butanol, isopropanol and the like. Among them, water can be suitably used.
  • the extraction can be performed under heating of the solvent (for example, about 85 to 105 ° C.). However, extraction can also be performed by sonication at lower temperatures (eg, 25-50 ° C., preferably 30-45 ° C.).
  • the “Shitake mushroom mycelium extract” is not limited, but for example, those obtained by the following method can be used. That is, shiitake mushrooms are inoculated on a solid medium based on bagasse (sugar cane pulp) and defatted rice bran. Next, the solid medium containing the shiitake mushroom mycelium obtained by growing the shiitake mushroom mycelium is loosened so that the amount of 12-mesh passage is 30% by weight or less. Water is added to the loosened solid medium, and the solid medium is crushed and ground while maintaining the temperature at 30 to 55 ° C., so that at least 70% by weight or more of the bagasse fiber passes through 12 mesh. Next, the temperature of the treated product is raised to 80 ° C., and the resulting suspension is filtered to obtain a shiitake mushroom mycelium extract.
  • the extract thus obtained may be used as it is as a Shiitake mushroom mycelium extract.
  • it may be stored as a powder by concentration, freeze drying or spray drying.
  • the powder thus prepared is useful because it can be used in various forms at the time of use.
  • the shiitake mushroom mycelium extract thus obtained contains 15 to 50%, preferably 20 to 40% (w / w) of saccharide by saccharide analysis by the phenol-sulfuric acid method. In addition, it contains 10 to 40%, preferably 13 to 30% (w / w) of the protein by the protein analysis by the Lowry method. Further, it contains 1 to 5%, preferably 2.5 to 3.5% (w / w) of polyphenols by the Folin-Denis method using gallic acid as a standard. The shiitake mushroom mycelium extract also contains about 0.1% lipid, about 0.4% fiber, about 20% ash, and the like.
  • composition % by mass
  • the composition may vary depending on the culture conditions and the like: xylose: 15.2; arabinose: 8.2; mannose: 8.4; glucose: 39.4; galactose: 5.4; amino sugar (glucosamine): 12 0.0; uronic acid: 11.3.
  • the composition for preventing or treating diabetic peripheral neuropathy according to the present invention comprises the above-mentioned Shiitake mushroom mycelium extract.
  • the content of the shiitake mushroom mycelium extract in the composition for preventing or treating diabetic peripheral neuropathy according to the present invention is not particularly limited.
  • the content of the Shiitake mushroom mycelium extract can be appropriately changed depending on the administration subject (including an animal) and the symptom (cause).
  • the composition for preventing or treating diabetic peripheral neuropathy contains the shiitake mushroom mycelium extract usually in an amount of 10% by mass or more, preferably 20% by mass or more, more preferably 30% by mass or more.
  • the content of Shiitake mushroom mycelium extract may be 100% by mass.
  • the composition for preventing or treating diabetic peripheral neuropathy according to the present invention can be provided as a therapeutic drug (pharmaceutical composition) for diabetic peripheral neuropathy.
  • the pharmaceutical composition according to the present invention can be provided as an internal preparation (oral preparation).
  • oral preparation for example, it can be provided in the form of a capsule, granule, powder, tablet, liquid, jelly or the like.
  • components such as a binder, a lubricant, a disintegrant, a colorant, a flavor, a preservative, an antioxidant, and a stabilizer.
  • antacids stomachic, digestive, intestinal, antispasmodic, mucosal repair, anti-inflammatory, astringent, antiemetic, antitussive, expectorant, anti-inflammatory enzyme, sedative-hypnotic, antihistamine, caffeine, inotropic diuresis
  • antibacterial agents vasoconstrictors, vasodilators, local anesthetics, crude drugs, herbal extract powders, vitamins, menthols, glucosamine compounds, chitin, chitosan and the like can be appropriately contained.
  • the pharmaceutical composition according to the present invention can be provided as an external preparation (transdermal preparation).
  • it can be provided in the form of a liquid, ointment, cream, gel, patch, aerosol, or the like.
  • components such as water, lower alcohol, polyhydric alcohol, solubilizer, surfactant, emulsion stabilizer, gelling agent, adhesive, stabilizer, pH adjuster, preservative, bactericide, fat, and oil can be contained.
  • an anti-inflammatory agent an analgesic, a local anesthetic, a vasodilator, a corticosteroid, a keratolytic agent, a humectant, a bactericide, an antioxidant, a refreshing agent, a fragrance, a pigment, and the like can be appropriately contained. .
  • the shiitake mushroom mycelium extract is usually contained in an amount of at least 10% by mass, preferably at least 20% by mass, more preferably at least 30% by mass, based on the total amount of the composition. Further, the pharmaceutical composition according to the present invention may have a shiitake mushroom mycelium extract content of 100% by mass.
  • composition for preventing or treating diabetic peripheral neuropathy according to the present invention can also be provided as a processed food.
  • Processed foods include, for example, candy, gum, jelly, biscuits, cookies, rice crackers, bread, noodles, fish and animal meat products, tea, soft drinks, coffee drinks, milk drinks, whey drinks, lactic acid bacteria drinks, yogurt, ice
  • General processed foods including taste foods such as creams and puddings and health foods can be exemplified.
  • the processed foods according to the present invention include functional health foods such as foods for specified health use and nutritional foods specified in the health functional food system of the Ministry of Health, Labor and Welfare. Further, it may contain dietary supplements (supplements), feeds, food additives, and the like.
  • the processed food according to the present invention can be prepared by including the shiitake mushroom mycelium extract in these processed foods.
  • the shiitake mushroom mycelium extract is generally contained in an amount of 10% by mass or more, preferably 20% by mass or more, more preferably 30% by mass or more. Further, the processed food according to the present invention may have a shiitake mushroom mycelium extract content of 100% by mass.
  • the dose of the composition for preventing or treating diabetic peripheral neuropathy according to the present invention can be appropriately selected depending on symptoms, age, body weight, elapsed time after onset, therapeutic measures used in combination, and the like.
  • the dose of the shiitake mycelium extract to a human adult as a dry powder weight is, for example, 4 mg / kg body weight or more, preferably 8 mg / kg body weight or more, more preferably 8 to 1700 mg / kg body weight per day. And more preferably 25 to 1700 mg / kg body weight.
  • the composition for preventing or treating diabetic peripheral neuropathy according to the present invention was subjected to 50 mg / kg body weight, 100 mg / kg body weight, 500 mg / kg per day by experiments using model mice. Body weight, 1000 mg / kg body weight, and 2000 mg / kg body weight are indicated. Such dosages are based on a human equivalent dose (HED) of 12.3 in mice ("Guidance for Industry ⁇ Estimating ⁇ the ⁇ Maximum ⁇ Safe ⁇ Starting ⁇ Dose ⁇ in ⁇ Initial ⁇ ClinicalTrials ⁇ for ⁇ Therapeutic.com).
  • HED human equivalent dose
  • composition for preventing or treating diabetic peripheral neuropathy according to the present invention can be used for animals such as domestic animals and companion animals in addition to humans.
  • the present invention provides a method for preventing, ameliorating, or treating the symptoms (administering a person by surgery or treatment) by administering a shiitake mushroom mycelium extract to a diabetic peripheral neuropathy patient or a subject to be administered as a preliminary group thereof. , Except for the method of diagnosis).
  • ⁇ Culture of Shiitake mycelium> As a medium for cultivating Shiitake mushroom mycelium, a solid medium comprising 80 parts by weight of bagasse and 10 parts by weight of defatted rice bran to which a suitable amount of pure water was added was used. Shiitake mushroom mycelium was inoculated into this medium, and allowed to stand in a culture room where the temperature and humidity were controlled to grow the shiitake mushroom mycelium.
  • Example 1 ⁇ Preventive effect on streptozotocin-induced diabetic mouse peripheral neuropathy>
  • the preventive effect of Shiitake mushroom mycelium extract on hyperesthesia such as allodynia due to mechanical stimuli induced by diabetes (usually severe pain caused by sensory stimuli that do not cause pain) and abnormal sensation in cold stimuli were examined.
  • a shiitake mushroom mycelium extract was orally administered to a mouse as a test substance, and the following tests (Cold plate test and von Frey test) were performed.
  • test substance administration group Using C57BL / 6J male mice of 6 to 7 weeks of age, control group, streptozotocin administration group, streptozotocin and test substance administration group (streptozotocin + test substance administration group) were divided into three groups. Configured.
  • the horizontal axis is the elapsed period (days) after administration, and the vertical axis is the escape reaction time (seconds). Those that can be judged to be significant from the control by a test with a significance level of 5% are marked with “*”. Similarly, in the following graphs, those for which the difference from the control can be determined to be significant by the test at the significance level of 5% are marked with “*”.
  • a white circle (written as “vehicle”), a vertical bar (written as “50 mg / kg LEM”), an asterisk (written as “100 mg / kg LEM”), and a white diamond. (Described as “500 mg / kg LEM”), a white triangle (described as “1000 mg / kg LEM”), and a white square (described as “2000 mg / kg LEM”) are control groups.
  • “Vehicle” is a mouse to which nothing was administered, and “50 mg / kg LEM”, “100 mg / kg LEM”, “500 mg / kg LEM”, “1000 mg / kg LEM”, and “2000 mg / kg LEM” were each treated with a test subject. It means that 50 mg / kg body weight, 100 mg / kg body weight, 500 mg / kg body weight, 1000 mg / kg body weight and 2000 mg / kg body weight were given per day. That is, these show the results when healthy mice were administered the test article.
  • Black circles (described as “200 mg / kg STZ”) are streptozotocin-administered groups. The result of giving 200 mg / kg body weight per day of streptozotocin is shown. Mice suffer from diabetes after administration of streptozotocin.
  • a black cross 50 mg / kg LEM + 200 mg / kg STZ (administered LEM from day 0)), a two-dot chain line only (100 mg / kg LEM + 200 mg / kg STZ (administered LEM from day 0)) and a black diamond (500 mg / kg LEM + 200 mg / kg STZ ( The black triangle (1000 mg / kg LEM + 200 mg / kg STZ (administered LEM from day 0)) and the black square (2000 mg / kg LEM + 200 mg / kg STZ (administered LEM from day 0)) were composed of streptozotocin and It is a test group.
  • the horizontal axis is the elapsed period (days) after administration, and the vertical axis is the avoidance response (score). If the number of avoidances is large, it is considered that the stimulation by the filament is more repelled.
  • the line types in FIG. 2 are the same as those in FIG. However, the vertical bar (shown as “50 mg / kg LEM” in FIG. 1) and the black cross (50 mg / kg LEM + 200 mg / kg STZ in FIG. 1 (administered LEM from day 0)) are not shown in FIG. In the control group, the avoidance response score was 1 or less stably.
  • test substance extract of Shiitake mushroom mycelium
  • streptozotocin has an effect of preventing diabetic peripheral neuropathy caused by streptozotocin.
  • Example 2 ⁇ Therapeutic effect on streptozotocin-induced diabetic mouse peripheral neuropathy> The therapeutic effect of Shiitake mushroom mycelium extract on hyperesthesia such as allodynia caused by mechanical stimulation induced by diabetes and paresthesia caused by low-temperature stimulation was examined. From day 21 after the administration of streptozotocin, a shiitake mushroom mycelium extract was orally administered to a mouse as a test substance, and the following tests (Cold plate test and von Frey test) were performed.
  • test substance Using C57BL / 6J male mice 6 to 7 weeks old, a control group, a streptozotocin administration group, a streptozotocin and a test substance administration group (streptozotocin + test substance administration group; (Administration started on day 3).
  • the horizontal axis is the elapsed period after administration (days), and the vertical axis is the escape reaction time (seconds).
  • the line types in the graph are the same as those in FIG. 2 for the control group and the streptozotocin-administered group.
  • “administration from day 0” in FIG. 2 was replaced with “administration from day 21” in FIG.
  • a white circle (written as “vehicle”), an asterisk (written as “100 mg / kg LEM”), a white diamond (written as “500 mg / kg LEM”), and a white triangle (Indicated as “1000 mg / kg LEM”) and a white square (indicated as “2000 mg / kg LEM”) are control groups.
  • “Vehicle” is a mouse to which nothing is administered, and “100 mg / kg LEM”, “500 mg / kg LEM”, “1000 mg / kg LEM”, and “2000 mg / kg LEM” indicate that the test substance is 100 mg / kg body weight per day. , 500 mg / kg body weight, 1000 mg / kg body weight, and 2000 mg / kg body weight. That is, they show the results when healthy mice were administered the test article.
  • the streptozotocin-administered group was designated as "STZ”.
  • STZ The streptozotocin-administered group
  • the two-dot chain line 100 mg / kg LEM + 200 mg / kg STZ (administered LEM from day 21)
  • the black diamond 500 mg / kg LEM + 200 mg / kg STZ (from day 21)).
  • the control group had an escape reaction time of about 15 to 18 seconds.
  • the escape reaction time was shorter as the administration elapsed time was longer. The escape reaction time became constant in about 14 days.
  • the horizontal axis is the elapsed period (days) after administration, and the vertical axis is the avoidance response (score). If the number of avoidances is large, it is considered that the stimulation by the filament is more repelled.
  • the line types in FIG. 4 are the same as those in FIG. Further, the same symbols in FIG. 3 were used for the symbols in the graph.
  • the avoidance response score was 1 or less stably.
  • the avoidance response score became higher as the administration elapsed time became longer, and the avoidance response score became constant at about 14 days. Thus, it was considered that the administration of streptozotocin caused diabetic peripheral neuropathy.
  • the composition for preventing or treating diabetic peripheral neuropathy according to the present invention shows that peripheral neuropathy is improved and has a therapeutic effect. Is shown.
  • composition according to the present invention can be used for preventing or treating peripheral neuropathy induced by diabetes.

Abstract

To provide a composition for preventing or treating diabetic peripheral neuropathy. A composition for preventing or treating diabetic peripheral neuropathy, said composition being characterized by comprising a Lentinus edodes mycelia extract, makes it possible to prevent or treat diabetes-induced numbness of limbs, pain of limbs, decrease in deep tendon reflex, muscular weakness, allodynia, hyperalgesia, disordered skilled motor function of fingers, gait disorder, stumbling, falling, flexion disorders [difficulty or impossibility in seiza (sitting erect with one's legs folded under one), agura (sitting with one's legs crossed), yokozuwari (sitting with one's legs out to one side), sitting on a chair, etc.], limb paralysis, etc.

Description

糖尿病性末梢神経障害の予防又は治療用組成物Composition for preventing or treating diabetic peripheral neuropathy
　本発明は、糖尿病によって誘発される末梢神経障害（以下、糖尿病性末梢神経障害と記載することがある）を予防又は治療する組成物に関するものである。 (4) The present invention relates to a composition for preventing or treating diabetes-induced peripheral neuropathy (hereinafter, sometimes referred to as diabetic peripheral neuropathy).
　糖尿病性末梢神経障害は、糖尿病の合併症である。糖尿病の進行に伴い、知覚過敏又は知覚鈍麻等の末梢神経障害が生ずることが知られている。この末梢神経障害により、患者の日常生活の質が低下するため、大きな問題となっている。しかしながら、糖尿病性末梢神経障害に対して有効な治療法は確立されていないのが現状である。糖尿病性末梢神経障害を予防および／又は治療方法が求められている。 Diabetic peripheral neuropathy is a complication of diabetes. It is known that peripheral neuropathy such as hyperesthesia or hypoesthesia occurs as diabetes progresses. This peripheral neuropathy is a major problem because the quality of daily life of patients is reduced. However, at present, no effective treatment has been established for diabetic peripheral neuropathy. There is a need for a method for preventing and / or treating diabetic peripheral neuropathy.
　例えば、特許文献１には、ラクタム化合物が糖輸送増強作用剤として効果があり、糖尿病、糖尿病性末梢神経障害、糖尿病性腎症、糖尿病性大血管症、耐糖能異常、又は肥満症の予防および／又は治療薬として利用できることが記載されている。 For example, Patent Document 1 discloses that a lactam compound is effective as a sugar transport-enhancing agent, preventing diabetes, diabetic peripheral neuropathy, diabetic nephropathy, diabetic macroangiopathy, impaired glucose tolerance, or preventing obesity and And / or can be used as a therapeutic agent.
特開２００６－２１３７３２号公報JP 2006-213732 A
　特許文献１には、ラクタム化合物が糖輸送能力を有する点については、実施例を含め開示されているが、末梢神経障害についての具体的な記載はない。特許文献１は、血糖値を下げる効果があるという開示にとどまる。 Patent Document 1 discloses that a lactam compound has a sugar transporting ability, including Examples, but does not specifically describe peripheral neuropathy. Patent Document 1 is limited to the disclosure that it has an effect of lowering the blood sugar level.
　糖尿病によって誘発される末梢神経障害の原因はまだ明らかではない。糖尿病の治療では、まずは糖尿病の進行を止めることが重要とされている。したがって、末梢神経障害に対する直接の治療は行われない場合が多い。そして、一度発症した糖尿病性末梢神経障害は、特許文献１に記載されるように一時的に血糖値を下げても治癒することは少ない。そのため、末梢神経障害を患った患者は、長期にわたる糖尿病の治療の間、継続して手足の痛みに悩まされることになる。 原因 The cause of diabetes-induced peripheral neuropathy is not yet clear. In the treatment of diabetes, it is important to first stop the progress of diabetes. Therefore, there is often no direct treatment for peripheral neuropathy. The diabetic peripheral neuropathy that has once developed is rarely cured even if the blood glucose level is temporarily lowered as described in Patent Document 1. Thus, patients suffering from peripheral neuropathy will continue to suffer from limb pain during long-term diabetes treatment.
　また、末梢神経障害の対処療法として、三環系抗うつ薬、α２δリガンドであるプレガバリン、およびセロトニン・ノルアドレナリン再取り込み阻害剤であるデュロキセチン等が推奨されているが、副作用も無視できない。また、これらの薬剤は糖尿病性末梢神経障害の予防組成物として利用することはできない。 In addition, tricyclic antidepressants, pregabalin as an α 2 δ ligand, duloxetine as a serotonin / noradrenaline reuptake inhibitor, and the like have been recommended as treatments for peripheral neuropathy, but the side effects cannot be ignored. In addition, these drugs cannot be used as a composition for preventing diabetic peripheral neuropathy.
　本発明の目的は、糖尿病性末梢神経障害を予防および治療できる、末梢神経障害の予防又は治療用組成物を提供することである。 An object of the present invention is to provide a composition for preventing or treating diabetic peripheral neuropathy, which is capable of preventing and treating diabetic peripheral neuropathy.
　本発明者らは、上記の課題に鑑みて糖尿病性末梢神経障害を予防又は治療することができる素材を探索し、シイタケ菌糸体を見出した。そして、本発明者らは、シイタケ菌糸体には、糖尿病性末梢神経障害に対して優れた予防又は治療効果を示すことを明らかにし、本発明を完成させた。 In view of the above problems, the present inventors searched for a material capable of preventing or treating diabetic peripheral neuropathy, and found a shiitake mycelium. The present inventors have clarified that the shiitake mushroom mycelium exhibits an excellent preventive or therapeutic effect on diabetic peripheral neuropathy, and completed the present invention.
　すなわち、第１の発明は、シイタケ菌糸体の抽出物を含有する糖尿病性末梢神経障害の予防又は治療用組成物である。 That is, the first invention is a composition for preventing or treating diabetic peripheral neuropathy, comprising an extract of Shiitake mushroom mycelium.
　また、第２の発明は、シイタケ菌糸体の抽出物を含有する糖尿病性末梢神経障害の予防又は治療用医薬組成物である。 The second invention is a pharmaceutical composition for preventing or treating diabetic peripheral neuropathy, comprising an extract of Shiitake mushroom mycelium.
　また、第２の発明はさらに、前記シイタケ菌糸体の抽出物の１日当たりの投与量が、乾燥粉末重量として４ｍｇ／ｋｇ体重以上である糖尿病性末梢神経障害の予防又は治療用医薬組成物である。 Further, the second invention is a pharmaceutical composition for preventing or treating diabetic peripheral neuropathy, wherein the daily dose of the extract of Shiitake mushroom mycelium is 4 mg / kg body weight or more as a dry powder weight. .
　また、第３の発明は、シイタケ菌糸体の抽出物を含有する糖尿病性末梢神経障害の予防又は治療効果を有する加工食品である。 The third invention is a processed food having an effect of preventing or treating diabetic peripheral neuropathy, comprising an extract of Shiitake mushroom mycelium.
　また、第４の発明は、第２の発明に係る糖尿病性末梢神経障害の予防又は治療用医薬組成物を対象に投与することを含む、糖尿病性末梢神経障害を予防又は治療する方法（人を手術・治療・診断する方法を除く）である。 In addition, a fourth invention provides a method for preventing or treating diabetic peripheral neuropathy, comprising administering to a subject the pharmaceutical composition for preventing or treating diabetic peripheral neuropathy according to the second invention. Excluding methods of surgery, treatment, and diagnosis).
　なお、本発明は、シイタケ菌糸体抽出物を用いた糖尿病性末梢神経障害の予防又は治療といってもよい。 Note that the present invention may also be referred to as prevention or treatment of diabetic peripheral neuropathy using a Shiitake mushroom mycelium extract.
　本発明は、糖尿病性末梢神経障害を予防又は治療することができる。すなわち、シイタケ菌糸体抽出物を投与することで、糖尿病によって誘発される四肢のしびれ、四肢の痛み、深部腱反射の低下、筋力の低下、アロディニア、痛覚過敏、手指の巧緻機能障害、歩行障害、躓き、転倒、屈曲障害（正座、あぐら、横座り又は椅子座り等の困難又は不能）、又は四肢の麻痺等が改善される。 The present invention can prevent or treat diabetic peripheral neuropathy. That is, by administering the Shiitake mushroom mycelium extract, numbness of the limbs induced by diabetes, pain in the limbs, decreased deep tendon reflex, decreased muscular strength, allodynia, hyperalgesia, elaborate dysfunction of the fingers, walking disorders, The stumbling, falling, bending disorder (difficult or impossible such as sitting straight, cross-legged, sitting sideways or sitting on a chair), or paralysis of limbs are improved.
　また、本発明に係る糖尿病性末梢神経障害の予防又は治療用組成物は経口摂取することで効果を奏することができる。したがって、通院する必要がなく、患者への負担が少ない。そして、糖尿病性末梢神経障害が改善されることによって患者の生活の質も向上する。 組成 In addition, the composition for preventing or treating diabetic peripheral neuropathy according to the present invention can exert its effects when taken orally. Therefore, there is no need to go to the hospital and the burden on the patient is small. The improvement of diabetic peripheral neuropathy also improves the quality of life of the patient.
　また、本発明に係る糖尿病性末梢神経障害の予防又は治療用組成物は、糖尿病性末梢神経障害の予防効果を奏するので、日常の食品と共に摂取することで、糖尿病性末梢神経障害の予防にも効果がある。 Further, the composition for preventing or treating diabetic peripheral neuropathy according to the present invention exerts a preventive effect on diabetic peripheral neuropathy, so that when taken together with daily food, it can also be used to prevent diabetic peripheral neuropathy. effective.
本発明に係る糖尿病性末梢神経障害の予防又は治療用組成物をストレプトゾトシンと共に投与した場合のコールドプレート試験の結果を示す図である。It is a figure which shows the result of the cold plate test when the composition for prevention or treatment of diabetic peripheral neuropathy according to the present invention is administered together with streptozotocin. 本発明に係る糖尿病性末梢神経障害の予防又は治療用組成物をストレプトゾトシンと共に投与した場合のフォン・フライ試験の結果を示す図である。It is a figure which shows the result of the von Frey test when the composition for prevention or treatment of diabetic peripheral neuropathy according to the present invention is administered together with streptozotocin. 本発明に係る糖尿病性末梢神経障害の予防又は治療用組成物をストレプトゾトシン投与後２１日目から投与した場合のコールドプレート試験の結果を示す図である。It is a figure which shows the result of the cold plate test when the composition for prevention or treatment of diabetic peripheral neuropathy according to the present invention is administered from day 21 after administration of streptozotocin. 本発明に係る糖尿病性末梢神経障害の予防又は治療用組成物をストレプトゾトシン投与後２１日目から投与した場合のフォン・フライ試験の結果を示す図である。It is a figure which shows the result of the von Frey test when the composition for prevention or treatment of diabetic peripheral neuropathy according to the present invention is administered from day 21 after administration of streptozotocin.
　以下に本発明に係る糖尿病性末梢神経障害の予防又は治療用組成物について図面および実施例を示し説明を行う。なお、以下の説明は、本発明の一実施形態および一実施例を例示するものであり、本発明が以下の説明に限定されるものではない。以下の説明は本発明の趣旨を逸脱しない範囲で改変することができる。 に つ い て Hereinafter, the composition for preventing or treating diabetic peripheral neuropathy according to the present invention will be described with reference to the drawings and examples. Note that the following description exemplifies one embodiment and one example of the present invention, and the present invention is not limited to the following description. The following description can be modified without departing from the spirit of the present invention.
　本発明において「予防」とは、未然防止、早期治療、機能低下防止の概念が含まれる。また、「治療」とは必ずしも罹患者の症状改善に限定されるものではなく、健常者（予備群）の症状の緩和や改善も含まれる概念である。 に お い て In the present invention, “prevention” includes the concepts of prevention, early treatment, and prevention of functional decline. In addition, “treatment” is not necessarily limited to the improvement of the symptoms of the affected patient, but is a concept that also includes the alleviation and improvement of the symptoms of a healthy person (preliminary group).
　本発明に係る糖尿病性末梢神経障害の予防又は治療用組成物において使用されるシイタケ菌糸体抽出物の原料となる「シイタケ菌糸体」（Ｌｅｎｔｉｎｕｓ　ｅｄｏｄｅｓ　ｍｙｃｅｌｉｕｍ）は、シイタケ菌糸体であれば特に限定はされない。例えば食用にされる子実体の前段階の菌糸の状態のものを使用することができる。本発明においては、例えば、シイタケ菌（Ｌｅｎｔｉｎｕｓ　ｅｄｏｄｅｓ）を固体培地で培養して得られる抽出物（シイタケ菌糸体抽出物）を用いることができる。 “Lentinus edodes mycelium”, which is a raw material of the shiitake mushroom mycelium extract used in the composition for preventing or treating diabetic peripheral neuropathy according to the present invention, is not particularly limited as long as it is a shiitake mushroom mycelium. Not done. For example, a mycelium in a stage before the edible fruiting body can be used. In the present invention, for example, an extract (Lentinus edodes mycelium extract) obtained by culturing Lentinus edodes on a solid medium can be used.
　本発明に係る糖尿病性末梢神経障害の予防又は治療用組成物において用いられるシイタケ菌糸体抽出物は、当該技術分野において公知の方法により調製することができる。例えば、シイタケ菌糸体を粉砕後に抽出して得られる抽出物は、シイタケ菌糸体抽出物として好適に用いることができる。さらに、例えば、シイタケ菌糸体を含む固体培地を水の存在下で粉砕、分解して得られる抽出物も、用いることができる。 The Shiitake mushroom mycelium extract used in the composition for preventing or treating diabetic peripheral neuropathy according to the present invention can be prepared by a method known in the art. For example, an extract obtained by extracting a shiitake mycelium after pulverization can be suitably used as a shiitake mushroom mycelium extract. Further, for example, an extract obtained by pulverizing and decomposing a solid medium containing Shiitake mycelium in the presence of water can also be used.
　抽出物の調製に用いられる溶媒としては、例えば、水、エタノール、メタノール、ブタノール、イソプロパノールなどを例示できる。なかでも水は、好適に利用することができる。抽出は溶媒の加熱下（例えば、８５～１０５℃程度）で行うこともできる。しかし、抽出は、より低温（例えば、２５～５０℃、好ましくは３０～４５℃）で超音波処理により行うこともできる。 溶媒 Examples of the solvent used for the preparation of the extract include water, ethanol, methanol, butanol, isopropanol and the like. Among them, water can be suitably used. The extraction can be performed under heating of the solvent (for example, about 85 to 105 ° C.). However, extraction can also be performed by sonication at lower temperatures (eg, 25-50 ° C., preferably 30-45 ° C.).
　この「シイタケ菌糸体抽出物」は、限定されるものではないが、例えば以下の方法により得られたものを使用することができる。すなわち、バガス（サトウキビのしぼりかす）と脱脂米糠を基材とする固体培地上にシイタケ菌を接種する。次いでシイタケ菌糸体を増殖させて得られるシイタケ菌糸体を含む固体培地を、１２メッシュ通過分が３０重量％以下となるようほぐす。ほぐした固体培地に水を添加し、３０～５５℃の温度に保ちながら前記固体培地を粉砕、すりつぶしてバガス繊維の少なくとも７０重量％以上が１２メッシュ通過分であるようにする。次いでその処理物の温度を８０℃にまで上昇させたのち、得られた懸濁状液をろ過することによってシイタケ菌糸体抽出液を得る。 The “Shitake mushroom mycelium extract” is not limited, but for example, those obtained by the following method can be used. That is, shiitake mushrooms are inoculated on a solid medium based on bagasse (sugar cane pulp) and defatted rice bran. Next, the solid medium containing the shiitake mushroom mycelium obtained by growing the shiitake mushroom mycelium is loosened so that the amount of 12-mesh passage is 30% by weight or less. Water is added to the loosened solid medium, and the solid medium is crushed and ground while maintaining the temperature at 30 to 55 ° C., so that at least 70% by weight or more of the bagasse fiber passes through 12 mesh. Next, the temperature of the treated product is raised to 80 ° C., and the resulting suspension is filtered to obtain a shiitake mushroom mycelium extract.
　本発明においては、このようにして得られた抽出液をそのままシイタケ菌糸体抽出物として使用してもよい。しかし、これを濃縮・凍結乾燥やスプレードライにより粉末として保存してもよい。このようにして調製された粉末は、使用時に種々の形態で使用できるので、有用である。 In the present invention, the extract thus obtained may be used as it is as a Shiitake mushroom mycelium extract. However, it may be stored as a powder by concentration, freeze drying or spray drying. The powder thus prepared is useful because it can be used in various forms at the time of use.
　このようにして得られるシイタケ菌糸体抽出物はフェノール－硫酸法による糖質分析により糖質を１５～５０％、好ましくは２０～４０％（ｗ／ｗ）含む。また、Ｌｏｗｒｙ法によるタンパク質分析によりタンパク質を１０～４０％、好ましくは１３～３０％（ｗ／ｗ）含む。また、没食子酸を標準とするＦｏｌｉｎ－Ｄｅｎｉｓ法によりポリフェノール類を１～５％、好ましくは２．５～３．５％（ｗ／ｗ）含む。シイタケ菌糸体抽出物にはそのほかに脂質約０．１％、繊維約０．４％、灰分約２０％などを含む。 シ The shiitake mushroom mycelium extract thus obtained contains 15 to 50%, preferably 20 to 40% (w / w) of saccharide by saccharide analysis by the phenol-sulfuric acid method. In addition, it contains 10 to 40%, preferably 13 to 30% (w / w) of the protein by the protein analysis by the Lowry method. Further, it contains 1 to 5%, preferably 2.5 to 3.5% (w / w) of polyphenols by the Folin-Denis method using gallic acid as a standard. The shiitake mushroom mycelium extract also contains about 0.1% lipid, about 0.4% fiber, about 20% ash, and the like.
　また、上述のようにして得られるシイタケ菌糸体抽出物の構成糖組成（質量％）の一例は、以下の通りであった。なお、この組成は培養条件などによって変動しうる：キシロース：１５．２；アラビノース：８．２；マンノース：８．４；グルコース：３９．４；ガラクトース：５．４；アミノ糖（グルコサミン）：１２．０；ウロン酸：１１．３。 Further, an example of the constituent sugar composition (% by mass) of the shiitake mushroom mycelium extract obtained as described above was as follows. The composition may vary depending on the culture conditions and the like: xylose: 15.2; arabinose: 8.2; mannose: 8.4; glucose: 39.4; galactose: 5.4; amino sugar (glucosamine): 12 0.0; uronic acid: 11.3.
　本発明に係る糖尿病性末梢神経障害の予防又は治療用組成物は上述のシイタケ菌糸体抽出物を含む。本発明に係る糖尿病性末梢神経障害の予防又は治療用組成物中のシイタケ菌糸体抽出物の含有量は特に限定されない。シイタケ菌糸体抽出物の含有量は、投与対象者（動物を含む）と、症状（原因）によって適宜変更することができる。一例としては、糖尿病性末梢神経障害の予防又は治療用組成物中にシイタケ菌糸体抽出物は、通常１０質量％以上、好ましくは２０質量％以上、より好ましくは３０質量％以上含有される。また、本発明に係る糖尿病性末梢神経障害の予防又は治療用組成物は、シイタケ菌糸体抽出物の含有量が１００質量％であってもよい。 は The composition for preventing or treating diabetic peripheral neuropathy according to the present invention comprises the above-mentioned Shiitake mushroom mycelium extract. The content of the shiitake mushroom mycelium extract in the composition for preventing or treating diabetic peripheral neuropathy according to the present invention is not particularly limited. The content of the Shiitake mushroom mycelium extract can be appropriately changed depending on the administration subject (including an animal) and the symptom (cause). As an example, the composition for preventing or treating diabetic peripheral neuropathy contains the shiitake mushroom mycelium extract usually in an amount of 10% by mass or more, preferably 20% by mass or more, more preferably 30% by mass or more. In the composition for preventing or treating diabetic peripheral neuropathy according to the present invention, the content of Shiitake mushroom mycelium extract may be 100% by mass.
　本発明に係る糖尿病性末梢神経障害の予防又は治療用組成物は、糖尿病性末梢神経障害用治療薬（医薬組成物）として提供することが可能である。本発明に係る医薬組成物は、内用剤（経口投与製剤）として提供することができる。例えば、カプセル剤、顆粒剤、散剤、錠剤、液剤、ゼリー剤等に製剤化して提供されうる。その際、結合剤、滑沢剤、崩壊剤、着色剤、矯味剤、防腐剤、抗酸化剤、安定化剤といった成分を含有させることが可能である。また、制酸剤、健胃剤、消化剤、整腸剤、鎮痙剤、粘膜修復剤、抗炎症剤、収斂剤、鎮吐剤、鎮咳剤、去痰剤、消炎酵素剤、鎮静催眠剤、抗ヒスタミン剤、カフェイン類、強心利尿剤、抗菌剤、血管収縮剤、血管拡張剤、局所麻酔剤、生薬、生薬エキス末、ビタミン類、メントール類、グルコサミン化合物、キチン、キトサン等を適宜含有させることができる。 The composition for preventing or treating diabetic peripheral neuropathy according to the present invention can be provided as a therapeutic drug (pharmaceutical composition) for diabetic peripheral neuropathy. The pharmaceutical composition according to the present invention can be provided as an internal preparation (oral preparation). For example, it can be provided in the form of a capsule, granule, powder, tablet, liquid, jelly or the like. At that time, it is possible to contain components such as a binder, a lubricant, a disintegrant, a colorant, a flavor, a preservative, an antioxidant, and a stabilizer. In addition, antacids, stomachic, digestive, intestinal, antispasmodic, mucosal repair, anti-inflammatory, astringent, antiemetic, antitussive, expectorant, anti-inflammatory enzyme, sedative-hypnotic, antihistamine, caffeine, inotropic diuresis Agents, antibacterial agents, vasoconstrictors, vasodilators, local anesthetics, crude drugs, herbal extract powders, vitamins, menthols, glucosamine compounds, chitin, chitosan and the like can be appropriately contained.
　さらに、本発明に係る医薬組成物は、外用剤（経皮投与製剤）として提供することができる。例えば、液剤、軟膏剤、クリーム剤、ゲル剤、貼付剤、エアゾール剤等に製剤化して提供されうる。その際、水、低級アルコール、多価アルコール、溶解補助剤、界面活性剤、乳化安定剤、ゲル化剤、粘着剤、安定化剤、ｐＨ調整剤、防腐剤、殺菌剤、脂、油といった成分を含有させることができる。また、抗炎症剤、鎮痛剤、局所麻酔剤、血管膨張剤、副腎皮質ホルモン、角質溶解剤、保湿剤、殺菌剤、抗酸化剤、清涼化剤、香料、色素などを適宜含有させることができる。 Furthermore, the pharmaceutical composition according to the present invention can be provided as an external preparation (transdermal preparation). For example, it can be provided in the form of a liquid, ointment, cream, gel, patch, aerosol, or the like. At that time, components such as water, lower alcohol, polyhydric alcohol, solubilizer, surfactant, emulsion stabilizer, gelling agent, adhesive, stabilizer, pH adjuster, preservative, bactericide, fat, and oil Can be contained. In addition, an anti-inflammatory agent, an analgesic, a local anesthetic, a vasodilator, a corticosteroid, a keratolytic agent, a humectant, a bactericide, an antioxidant, a refreshing agent, a fragrance, a pigment, and the like can be appropriately contained. .
　なお、本発明に係る医薬組成物において、シイタケ菌糸体抽出物は、組成物全量に対して通常１０質量％以上、好ましくは２０質量％以上、より好ましくは３０質量％以上含有される。また、本発明に係る医薬組成物は、シイタケ菌糸体抽出物の含有量が１００質量％であってもよい。 In the pharmaceutical composition according to the present invention, the shiitake mushroom mycelium extract is usually contained in an amount of at least 10% by mass, preferably at least 20% by mass, more preferably at least 30% by mass, based on the total amount of the composition. Further, the pharmaceutical composition according to the present invention may have a shiitake mushroom mycelium extract content of 100% by mass.
　また、本発明に係る糖尿病性末梢神経障害の予防又は治療用組成物は加工食品として提供することも可能である。 The composition for preventing or treating diabetic peripheral neuropathy according to the present invention can also be provided as a processed food.
　加工食品としては、例えば、飴、ガム、ゼリー、ビスケット、クッキー、煎餅、パン、麺、魚肉・畜肉練製品、茶、清涼飲料、コーヒー飲料、乳飲料、乳清飲料、乳酸菌飲料、ヨーグルト、アイスクリーム、プリン等といった嗜好食品や健康食品を含む一般加工食品が例示できる。また、これらの一般加工食品だけでなく、本発明に係る加工食品は、厚生労働省の保健機能食品制度に規定された特定保健用食品や栄養機能食品などの保健機能食品を含む。さらに、栄養補助食品（サプリメント）、飼料、食品添加物等を含んでもよい。 Processed foods include, for example, candy, gum, jelly, biscuits, cookies, rice crackers, bread, noodles, fish and animal meat products, tea, soft drinks, coffee drinks, milk drinks, whey drinks, lactic acid bacteria drinks, yogurt, ice General processed foods including taste foods such as creams and puddings and health foods can be exemplified. In addition to these general processed foods, the processed foods according to the present invention include functional health foods such as foods for specified health use and nutritional foods specified in the health functional food system of the Ministry of Health, Labor and Welfare. Further, it may contain dietary supplements (supplements), feeds, food additives, and the like.
　これらの加工食品に、シイタケ菌糸体抽出物を含有することで本発明に係る加工食品を調製することができる。なお、本発明に係る加工食品において、シイタケ菌糸体抽出物は、通常１０質量％以上、好ましくは２０質量％以上、より好ましくは３０質量％以上含有される。また、本発明に係る加工食品は、シイタケ菌糸体抽出物の含有量が１００質量％であってもよい。 加工 The processed food according to the present invention can be prepared by including the shiitake mushroom mycelium extract in these processed foods. In the processed food according to the present invention, the shiitake mushroom mycelium extract is generally contained in an amount of 10% by mass or more, preferably 20% by mass or more, more preferably 30% by mass or more. Further, the processed food according to the present invention may have a shiitake mushroom mycelium extract content of 100% by mass.
　本発明に係る糖尿病性末梢神経障害の予防又は治療用組成物の投与量は、症状、年齢、体重、発症後の経過時間、併用される治療的措置などにより、適宜選択することができる。シイタケ菌糸体抽出物の乾燥粉末重量としてヒト成人への投与量は、例えば１日あたり、４ｍｇ／ｋｇ体重以上であり、好ましくは８ｍｇ／ｋｇ体重以上であり、より好ましくは８～１７００ｍｇ／ｋｇ体重であり、更に好ましくは２５～１７００ｍｇ／ｋｇ体重である。 投 与 The dose of the composition for preventing or treating diabetic peripheral neuropathy according to the present invention can be appropriately selected depending on symptoms, age, body weight, elapsed time after onset, therapeutic measures used in combination, and the like. The dose of the shiitake mycelium extract to a human adult as a dry powder weight is, for example, 4 mg / kg body weight or more, preferably 8 mg / kg body weight or more, more preferably 8 to 1700 mg / kg body weight per day. And more preferably 25 to 1700 mg / kg body weight.
　後述する本実施例において、モデルマウスを用いた実験により、本発明に係る糖尿病性末梢神経障害の予防又は治療用組成物を、１日あたり、５０ｍｇ／ｋｇ体重、１００ｍｇ／ｋｇ体重、５００ｍｇ／ｋｇ体重、１０００ｍｇ／ｋｇ体重、および２０００ｍｇ／ｋｇ体重投与したことが示されている。かかる投与量は、マウスにおけるヒト等価用量（ＨＥＤ）１２．３（「Ｇｕｉｄａｎｃｅ　ｆｏｒＩｎｄｕｓｔｒｙ　Ｅｓｔｉｍａｔｉｎｇ　ｔｈｅ　Ｍａｘｉｍｕｍ　Ｓａｆｅ　Ｓｔａｒｔｉｎｇ　Ｄｏｓｅ　ｉｎ　Ｉｎｉｔｉａｌ　ＣｌｉｎｉｃａｌＴｒｉａｌｓ　ｆｏｒ　Ｔｈｅｒａｐｅｕｔｉｃｓ　ｉｎ　Ａｄｕｌｔ　Ｈｅａｌｔｈｙ　Ｖｏｌｕｎｔｅｅｒｓ」参照）に基づき、ヒト成人（体重６０ｋｇ）への１日あたりの投与量に換算した場合、それぞれ、約４ｍｇ／ｋｇ体重、約８ｍｇ／ｋｇ体重、約４１ｍｇ／ｋｇ体重、約８１ｍｇ／ｋｇ体重、約１６３ｍｇ／ｋｇ体重である。この変換は、より一般的な式ＨＥＤ＝動物用量ｍｇ／ｋｇ×（動物体重（ｋｇ）／ヒト体重（ｋｇ））０．３３に基づく。 In the below-described Examples, the composition for preventing or treating diabetic peripheral neuropathy according to the present invention was subjected to 50 mg / kg body weight, 100 mg / kg body weight, 500 mg / kg per day by experiments using model mice. Body weight, 1000 mg / kg body weight, and 2000 mg / kg body weight are indicated. Such dosages are based on a human equivalent dose (HED) of 12.3 in mice ("Guidance for Industry \ Estimating \ the \ Maximum \ Safe \ Starting \ Dose \ in \ Initial \ ClinicalTrials \ for \ Therapeutic.com). When converted to daily doses, they are about 4 mg / kg body weight, about 8 mg / kg body weight, about 41 mg / kg body weight, about 81 mg / kg body weight, and about 163 mg / kg body weight, respectively. This conversion is based on the more general formula HED = animal dose mg / kg x (animal body weight (kg) / human body weight (kg)) 0.33.
　本発明に係る糖尿病性末梢神経障害の予防又は治療用組成物は、ヒトの他、家畜・愛玩動物などの動物用に使用することもできる。 組成 The composition for preventing or treating diabetic peripheral neuropathy according to the present invention can be used for animals such as domestic animals and companion animals in addition to humans.
　なお、本発明は、シイタケ菌糸体抽出物を、糖尿病性末梢神経障害患者又はその予備群である投与対象者に投与することで、当該症状を予防、改善又は治療する方法（人を手術、治療、診断する方法を除く）であるといってよい。 The present invention provides a method for preventing, ameliorating, or treating the symptoms (administering a person by surgery or treatment) by administering a shiitake mushroom mycelium extract to a diabetic peripheral neuropathy patient or a subject to be administered as a preliminary group thereof. , Except for the method of diagnosis).
　＜シイタケ菌糸体の培養＞
　シイタケ菌糸体の培養を行う培地は、バガス８０重量部、脱脂米糠１０重量部からなる固体培地に適度な純水を加えたものを用いた。この培地にシイタケ菌糸体を接種し、温度および湿度を管理した培養室内に放置し、シイタケ菌糸体を増殖させた。 <Culture of Shiitake mycelium>
As a medium for cultivating Shiitake mushroom mycelium, a solid medium comprising 80 parts by weight of bagasse and 10 parts by weight of defatted rice bran to which a suitable amount of pure water was added was used. Shiitake mushroom mycelium was inoculated into this medium, and allowed to stand in a culture room where the temperature and humidity were controlled to grow the shiitake mushroom mycelium.
　＜シイタケ菌糸体抽出物の乾燥粉末の製造＞
　シイタケ菌糸体が蔓延した培地に８等量の純水を加えて粉砕し、８０℃±３℃で２時間加熱を行い、液循環抽出をした後１００メッシュろ過した。ろ過抽出物をＢｒｉｘ値が２７％±２％となるように濃縮した。濃縮した抽出物をヘリコイド式滅菌機を用いて１３５℃１５秒で滅菌した。滅菌抽出物に滅菌水を加えてＢｒｉｘ値が２５％±１％となるように調整した後、凍結乾燥を行った。この凍結乾燥粉末を粉砕後４２メッシュ通過させ、シイタケ菌糸体の乾燥粉末を得た。これを以下の実施例で披検物（シイタケ菌糸体の抽出物）とした。 <Production of dry powder of shiitake mycelium extract>
8 equivalents of pure water was added to the medium in which Shiitake mushroom mycelium spread, and the mixture was pulverized, heated at 80 ° C. ± 3 ° C. for 2 hours, subjected to liquid circulation extraction, and then subjected to 100 mesh filtration. The filtered extract was concentrated to a Brix value of 27% ± 2%. The concentrated extract was sterilized at 135 ° C. for 15 seconds using a helicoid sterilizer. The Brix value was adjusted to 25% ± 1% by adding sterilized water to the sterilized extract, and then freeze-dried. This lyophilized powder was pulverized and passed through a 42 mesh to obtain a dried powder of Shiitake mycelium. This was used as a test substance (extract of shiitake mushroom mycelium) in the following examples.
　（実施例１）＜ストレプトゾトシン誘発糖尿病マウス末梢神経障害に対する予防効果＞
　糖尿病によって誘発される、機械的刺激によるアロディニア（通常痛みを引き起こさない感覚刺激で惹起される激痛）等の知覚過敏および低温刺激における知覚異常に対するシイタケ菌糸体抽出物の予防効果を調べた。ストレプトゾトシン投与と同時にシイタケ菌糸体抽出物を被検物としてマウスに経口投与し、以下の試験（Ｃｏｌｄ　ｐｌａｔｅ　ｔｅｓｔおよびｖｏｎ　Ｆｒｅｙ　ｔｅｓｔ）を行った。 (Example 1) <Preventive effect on streptozotocin-induced diabetic mouse peripheral neuropathy>
The preventive effect of Shiitake mushroom mycelium extract on hyperesthesia such as allodynia due to mechanical stimuli induced by diabetes (usually severe pain caused by sensory stimuli that do not cause pain) and abnormal sensation in cold stimuli were examined. Simultaneously with the administration of streptozotocin, a shiitake mushroom mycelium extract was orally administered to a mouse as a test substance, and the following tests (Cold plate test and von Frey test) were performed.
　（１）被検物の投与
　６～７週齢のＣ５７ＢＬ／６Ｊ雄性マウスを用い、コントロール群、ストレプトゾトシン投与群、ストレプトゾトシンおよび被検物投与群（ストレプトゾトシン＋被検物投与群）の３群に群構成した。 (1) Administration of test substance Using C57BL / 6J male mice of 6 to 7 weeks of age, control group, streptozotocin administration group, streptozotocin and test substance administration group (streptozotocin + test substance administration group) were divided into three groups. Configured.
　（２）コールドプレート試験（Ｃｏｌｄ　ｐｌａｔｅ　ｔｅｓｔ）
　コールドプレート試験を行い、低温刺激における知覚異常に対するシイタケ菌糸体抽出物の効果を試験した。上記（１）の３群のマウスを４℃に設定したコールドプレート上にのせ、逃避までの反応時間（潜時）を測定した。潜時が短いほどコールドプレートによる低温刺激を、より忌避していると考えられる。結果を図１に示す。 (2) Cold plate test
A cold plate test was performed to test the effect of shiitake mushroom mycelium extract on paresthesia in cold stimulation. The three groups of mice of the above (1) were placed on a cold plate set at 4 ° C., and the reaction time (latency) until escape was measured. It is considered that the shorter the latency, the more the cold plate stimulus due to the cold plate is repelled. The results are shown in FIG.
　図１の、横軸は投与後経過期間（日）であり、縦軸は逃避反応時間（秒）である。有意水準５％の検定でコントロールとの差が有意であると判断できるものには「＊」を付した。以下のグラフでも同様に有意水準５％の検定でコントロールとの差が有意であると判断できるものには「＊」を付した。 の In FIG. 1, the horizontal axis is the elapsed period (days) after administration, and the vertical axis is the escape reaction time (seconds). Those that can be judged to be significant from the control by a test with a significance level of 5% are marked with “*”. Similarly, in the following graphs, those for which the difference from the control can be determined to be significant by the test at the significance level of 5% are marked with “*”.
　グラフの線種において、白丸（「ｖｅｈｉｃｌｅ」と記した。）と、縦棒（「５０ｍｇ／ｋｇＬＥＭ」と記した。）と、米印（「１００ｍｇ／ｋｇＬＥＭ」と記した。）と、白菱形（「５００ｍｇ／ｋｇＬＥＭ」と記した。）と、白三角（「１０００ｍｇ／ｋｇＬＥＭ」と記した。）と、白四角（「２０００ｍｇ／ｋｇＬＥＭ」と記した。）は、コントロール群である。 In the line type of the graph, a white circle (written as “vehicle”), a vertical bar (written as “50 mg / kg LEM”), an asterisk (written as “100 mg / kg LEM”), and a white diamond. (Described as “500 mg / kg LEM”), a white triangle (described as “1000 mg / kg LEM”), and a white square (described as “2000 mg / kg LEM”) are control groups.
　「ｖｅｈｉｃｌｅ」は何も投与しないマウスであり、「５０ｍｇ／ｋｇＬＥＭ」、「１００ｍｇ／ｋｇＬＥＭ」、「５００ｍｇ／ｋｇＬＥＭ」、「１０００ｍｇ／ｋｇＬＥＭ」、「２０００ｍｇ／ｋｇＬＥＭ」は、それぞれ被検物を、１日あたり５０ｍｇ／ｋｇ体重、１００ｍｇ／ｋｇ体重、５００ｍｇ／ｋｇ体重、１０００ｍｇ／ｋｇ体重、２０００ｍｇ／ｋｇ体重与えたという意味である。つまり、これらは健康なマウスが被検物を投与された場合の結果を示す。 “Vehicle” is a mouse to which nothing was administered, and “50 mg / kg LEM”, “100 mg / kg LEM”, “500 mg / kg LEM”, “1000 mg / kg LEM”, and “2000 mg / kg LEM” were each treated with a test subject. It means that 50 mg / kg body weight, 100 mg / kg body weight, 500 mg / kg body weight, 1000 mg / kg body weight and 2000 mg / kg body weight were given per day. That is, these show the results when healthy mice were administered the test article.
　黒丸（「２００ｍｇ／ｋｇＳＴＺ」と記した。）は、ストレプトゾトシン投与群である。ストレプトゾトシンを、１日あたり２００ｍｇ／ｋｇ体重与えた結果を示す。ストレプトゾトシンの投与によって、マウスは糖尿病を患うことになる。 Black circles (described as “200 mg / kg STZ”) are streptozotocin-administered groups. The result of giving 200 mg / kg body weight per day of streptozotocin is shown. Mice suffer from diabetes after administration of streptozotocin.
　黒バツ印（５０ｍｇ／ｋｇＬＥＭ＋２００ｍｇ／ｋｇＳＴＺ（０日目よりＬＥＭ投与））と、二点鎖線のみ（１００ｍｇ／ｋｇＬＥＭ＋２００ｍｇ／ｋｇＳＴＺ（０日目よりＬＥＭ投与））と、黒菱形（５００ｍｇ／ｋｇＬＥＭ＋２００ｍｇ／ｋｇＳＴＺ（０日目よりＬＥＭ投与））と、黒三角（１０００ｍｇ／ｋｇＬＥＭ＋２００ｍｇ／ｋｇＳＴＺ（０日目よりＬＥＭ投与））と、黒四角（２０００ｍｇ／ｋｇＬＥＭ＋２００ｍｇ／ｋｇＳＴＺ（０日目よりＬＥＭ投与））は、ストレプトゾトシン及び被検物投与群である。これらは、一日あたり、それぞれストレプトゾトシン２００ｍｇ／ｋｇ体重と共に、被検物を５０ｍｇ／ｋｇ体重、１００ｍｇ／ｋｇ体重、５００ｍｇ／ｋｇ体重、１０００ｍｇ／ｋｇ体重、２０００ｍｇ／ｋｇ体重投与した結果を示す。 A black cross (50 mg / kg LEM + 200 mg / kg STZ (administered LEM from day 0)), a two-dot chain line only (100 mg / kg LEM + 200 mg / kg STZ (administered LEM from day 0)) and a black diamond (500 mg / kg LEM + 200 mg / kg STZ ( The black triangle (1000 mg / kg LEM + 200 mg / kg STZ (administered LEM from day 0)) and the black square (2000 mg / kg LEM + 200 mg / kg STZ (administered LEM from day 0)) were composed of streptozotocin and It is a test group. These show the results of administering 50 mg / kg body weight, 100 mg / kg body weight, 500 mg / kg body weight, 1000 mg / kg body weight, and 2000 mg / kg body weight of the test substance together with streptozotocin 200 mg / kg body weight per day.
　コントロール群は全て逃避反応時間が約１５秒から１８秒であり、安定していた。ストレプトゾトシン投与群は、６日目から逃避反応時間が短くなり、１４日目には、コールドプレートにおける冷刺激に対してストレプトゾトシン投与群（グラフ中「ＳＴＺ」と記した。）では著しく潜時が短縮された。なお、グラフ中ストレプトゾトシン２００ｍｇ／ｋｇ体重と共に被検物を５０ｍｇ／ｋｇ体重投与したマウスを「５０ＬＳ」と記した。 All control groups were stable with escape reaction times of about 15 to 18 seconds. In the streptozotocin-administered group, the escape reaction time was shortened from the sixth day, and on the 14th day, the latency was significantly shortened in the streptozotocin-administered group (denoted by “STZ” in the graph) in response to the cold stimulation on the cold plate. Was done. In addition, the mouse | mouth which administered the test substance with 50 mg / kg body weight with streptozotocin 200 mg / kg body weight in a graph was described as "50LS."
　（３）フォン・フライ試験（ｖｏｎ　Ｆｒｅｙ　ｔｅｓｔ）
　ケージに、上記（１）の３群のマウスを入れ、強度０．１６ｇのフィラメントを後肢裏に押し付けて回避反応の回数を測定した。ストレプトゾトシン投与群では、コントロール群に比べて著しく回避反応スコアが上昇し、ストレプトゾトシン＋被検物投与群では、コントロール群と同程度の回避反応スコアを示した。ストレプトゾトシン＋被検物投与群は、ストレプトゾトシン投与群と比較して有意に回避反応スコアの上昇を抑制した。結果を図２に示す。 (3) von Frey test
The three groups of mice described in (1) above were placed in a cage, and a filament having a strength of 0.16 g was pressed against the back of the hind limb to measure the number of avoidance reactions. In the streptozotocin-administered group, the avoidance response score was significantly increased as compared with the control group, and in the streptozotocin + test group, the avoidance response score was almost the same as that of the control group. The streptozotocin + test substance administration group significantly suppressed the increase in the avoidance response score as compared with the streptozotocin administration group. FIG. 2 shows the results.
　図２の、横軸は投与後経過期間（日）であり、縦軸は回避反応（スコア）である。回避回数が多ければ、フィラメントによる刺激をより忌避していると考えられる。図２中の線種については、図１の場合と同じである。ただし、縦棒（図１で「５０ｍｇ／ｋｇＬＥＭ」と記した。）と黒バツ印（図１で５０ｍｇ／ｋｇＬＥＭ＋２００ｍｇ／ｋｇＳＴＺ（０日目よりＬＥＭ投与））は図２にはない。コントロール群では、安定して回避反応スコアが１以下であった。 In FIG. 2, the horizontal axis is the elapsed period (days) after administration, and the vertical axis is the avoidance response (score). If the number of avoidances is large, it is considered that the stimulation by the filament is more repelled. The line types in FIG. 2 are the same as those in FIG. However, the vertical bar (shown as “50 mg / kg LEM” in FIG. 1) and the black cross (50 mg / kg LEM + 200 mg / kg STZ in FIG. 1 (administered LEM from day 0)) are not shown in FIG. In the control group, the avoidance response score was 1 or less stably.
　一方、ストレプトゾトシン投与群は、６日目から回避反応スコアが高くなった。ストレプトゾトシン＋被検物投与群は、ほとんどコントロール群と同じ回避反応スコアであった。 On the other hand, in the streptozotocin administration group, the avoidance response score increased from day 6. The streptozotocin + subject administration group had almost the same avoidance response score as the control group.
　以上のことより、被検物（シイタケ菌糸体の抽出物）は、ストレプトゾトシンによる糖尿病性末梢神経障害を予防する効果がある。 From the above, the test substance (extract of Shiitake mushroom mycelium) has an effect of preventing diabetic peripheral neuropathy caused by streptozotocin.
　（実施例２）＜ストレプトゾトシン誘発糖尿病マウス末梢神経障害に対する治療効果＞
　糖尿病よって誘発される、機械的刺激によるアロディニア等の知覚過敏および低温刺激における知覚異常に対するシイタケ菌糸体抽出物の治療効果を調べた。ストレプトゾトシン投与後２１日目からシイタケ菌糸体抽出物を被検物としてマウスに経口投与し、以下の試験（Ｃｏｌｄ　ｐｌａｔｅ　ｔｅｓｔおよびｖｏｎ　Ｆｒｅｙ　ｔｅｓｔ）を行った。 (Example 2) <Therapeutic effect on streptozotocin-induced diabetic mouse peripheral neuropathy>
The therapeutic effect of Shiitake mushroom mycelium extract on hyperesthesia such as allodynia caused by mechanical stimulation induced by diabetes and paresthesia caused by low-temperature stimulation was examined. From day 21 after the administration of streptozotocin, a shiitake mushroom mycelium extract was orally administered to a mouse as a test substance, and the following tests (Cold plate test and von Frey test) were performed.
　（１）被検物の投与
　６～７週齢のＣ５７ＢＬ／６Ｊ雄性マウスを用い、コントロール群、ストレプトゾトシン投与群、ストレプトゾトシンおよび被検物投与群（ストレプトゾトシン＋被検物投与群；被検物は２１日目から投与開始）の３群に群構成した。 (1) Administration of test substance Using C57BL / 6J male mice 6 to 7 weeks old, a control group, a streptozotocin administration group, a streptozotocin and a test substance administration group (streptozotocin + test substance administration group; (Administration started on day 3).
　（２）コールドプレート試験（Ｃｏｌｄ　ｐｌａｔｅ　ｔｅｓｔ）
　コールドプレート試験を行い、低温刺激における知覚異常に対するシイタケ菌糸体抽出物の効果を試験した。上記（１）の３群のマウスを４℃に設定したコールドプレート上にのせ、逃避までの反応時間（潜時）を測定した。潜時が短いほどコールドプレートによる低温刺激を、より忌避していると考えられる。結果を図３に示す。 (2) Cold plate test
A cold plate test was performed to test the effect of shiitake mushroom mycelium extract on paresthesia in cold stimulation. The three groups of mice of the above (1) were placed on a cold plate set at 4 ° C., and the reaction time (latency) until escape was measured. It is considered that the shorter the latency, the more the cold plate stimulus due to the cold plate is repelled. The results are shown in FIG.
　図３を参照して、横軸は投与後経過期間（日）であり、縦軸は逃避反応時間（秒）である。また、グラフ中の線種はコントロール群とストレプトゾトシン投与群は図２の場合と同じである。ストレプトゾトシン及び被検物投与群については、図２で「０日より投与」としたのを、図３では「２１日目より投与」に読み替えた。 を Referring to FIG. 3, the horizontal axis is the elapsed period after administration (days), and the vertical axis is the escape reaction time (seconds). The line types in the graph are the same as those in FIG. 2 for the control group and the streptozotocin-administered group. Regarding the streptozotocin and test substance administration groups, “administration from day 0” in FIG. 2 was replaced with “administration from day 21” in FIG.
　グラフの線種において、白丸（「ｖｅｈｉｃｌｅ」と記した。）と、米印（「１００ｍｇ／ｋｇＬＥＭ」と記した。）と、白菱形（「５００ｍｇ／ｋｇＬＥＭ」と記した。）と、白三角（「１０００ｍｇ／ｋｇＬＥＭ」と記した。）と、白四角（「２０００ｍｇ／ｋｇＬＥＭ」と記した。）は、コントロール群である。 In the line type of the graph, a white circle (written as “vehicle”), an asterisk (written as “100 mg / kg LEM”), a white diamond (written as “500 mg / kg LEM”), and a white triangle (Indicated as “1000 mg / kg LEM”) and a white square (indicated as “2000 mg / kg LEM”) are control groups.
　「ｖｅｈｉｃｌｅ」は何も投与しないマウスであり、「１００ｍｇ／ｋｇＬＥＭ」、「５００ｍｇ／ｋｇＬＥＭ」、「１０００ｍｇ／ｋｇＬＥＭ」、「２０００ｍｇ／ｋｇＬＥＭ」は、それぞれ被検物を、１日あたり１００ｍｇ／ｋｇ体重、５００ｍｇ／ｋｇ体重、１０００ｍｇ／ｋｇ体重、２０００ｍｇ／ｋｇ体重与えたという意味である。つまり、これらは健康なマウスが被検物を投与された場合の結果を示す。 “Vehicle” is a mouse to which nothing is administered, and “100 mg / kg LEM”, “500 mg / kg LEM”, “1000 mg / kg LEM”, and “2000 mg / kg LEM” indicate that the test substance is 100 mg / kg body weight per day. , 500 mg / kg body weight, 1000 mg / kg body weight, and 2000 mg / kg body weight. That is, they show the results when healthy mice were administered the test article.
　また、図３では、グラフ中に以下の記号を付した。ストレプトゾトシン投与群を「ＳＴＺ」と示した。また、ストレプトゾトシン及び被検物投与群については、二点鎖線のみ（１００ｍｇ／ｋｇＬＥＭ＋２００ｍｇ／ｋｇＳＴＺ（２１日目よりＬＥＭ投与））を「１００ＬＳ」とし、黒菱形（５００ｍｇ／ｋｇＬＥＭ＋２００ｍｇ／ｋｇＳＴＺ（２１日目よりＬＥＭ投与））を「５００ＬＳ」とし、黒三角（１０００ｍｇ／ｋｇＬＥＭ＋２００ｍｇ／ｋｇＳＴＺ（２１日目よりＬＥＭ投与））を「１０００ＬＳ」とし、黒四角（２０００ｍｇ／ｋｇＬＥＭ＋２００ｍｇ／ｋｇＳＴＺ（２１日目よりＬＥＭ投与））を「２０００ＬＳ」とした。 In FIG. 3, the following symbols are given in the graph. The streptozotocin-administered group was designated as "STZ". For the streptozotocin and test substance administration groups, only the two-dot chain line (100 mg / kg LEM + 200 mg / kg STZ (administered LEM from day 21)) was set to “100LS”, and the black diamond (500 mg / kg LEM + 200 mg / kg STZ (from day 21)). LEM)) is set to “500LS”, the black triangle (1000 mg / kg LEM + 200 mg / kg STZ (administered LEM from day 21)) is set to “1000LS”, and the black square (2000 mg / kg LEM + 200 mg / kg STZ (administered LEM from day 21)) Was set to "2000LS".
　図３を参照して、コントロール群は約１５秒から１８秒程度の逃避反応時間であった。一方、ストレプトゾトシン＋被検物投与群およびストレプトゾトシン投与群は投与経過期間が延びるほど逃避反応時間が短くなった。そしてほぼ１４日で逃避反応時間は一定となった。 を Referring to FIG. 3, the control group had an escape reaction time of about 15 to 18 seconds. On the other hand, in the streptozotocin + test substance administration group and the streptozotocin administration group, the escape reaction time was shorter as the administration elapsed time was longer. The escape reaction time became constant in about 14 days.
　２１日目に被検物の投与を開始し（図３中では、「ＬＥＭ投与」と示した。）、この日よりストレプトゾトシン＋被検物投与群は、逃避反応時間が延びた。 投 与 Administration of the test substance was started on day 21 (indicated as “LEM administration” in FIG. 3), and from that day on, the escape reaction time of the streptozotocin + test substance administration group was prolonged.
　（３）フォン・フライ試験（ｖｏｎ　Ｆｒｅｙ　ｔｅｓｔ）
　ケージに、上記（１）の３群のマウスを入れ、強度０．１６ｇのフィラメントを後肢裏に押し付けて回避反応の回数を測定した。ストレプトゾトシン投与群では、コントロール群に比べて著しく回避反応スコアが上昇した。ストレプトゾトシン＋被検物投与群では、被検物を投与しない間は、ストレプトゾトシン投与群同様に、回避反応スコアは上昇した。しかし、投与日以後は、回避反応スコアは減少し、コントロール群と同程度の回避反応スコアまで回復した。ストレプトゾトシン＋被検物投与群は、ストレプトゾトシン投与群と比較して有意に回避反応スコアの上昇を抑制した。結果を図４に示す。 (3) von Frey test
The three groups of mice described in (1) above were placed in a cage, and a filament having a strength of 0.16 g was pressed against the back of the hind limb to measure the number of avoidance reactions. The avoidance response score was significantly increased in the streptozotocin-administered group compared to the control group. In the streptozotocin + test substance administration group, the avoidance response score increased while the test substance was not administered, as in the streptozotocin administration group. However, after the administration day, the avoidance response score decreased and recovered to the same level as that of the control group. The streptozotocin + test substance administration group significantly suppressed the increase in the avoidance response score as compared with the streptozotocin administration group. FIG. 4 shows the results.
　図４を参照して、横軸は投与後経過期間（日）であり、縦軸は回避反応（スコア）である。回避回数が多ければ、フィラメントによる刺激をより忌避していると考えられる。図４中の線種については、図３の場合と同じである。また、グラフ中の記号についても図３と同じものを用いた。 を Referring to FIG. 4, the horizontal axis is the elapsed period (days) after administration, and the vertical axis is the avoidance response (score). If the number of avoidances is large, it is considered that the stimulation by the filament is more repelled. The line types in FIG. 4 are the same as those in FIG. Further, the same symbols in FIG. 3 were used for the symbols in the graph.
　コントロール群では、安定して回避反応スコアが１以下であった。一方、ストレプトゾトシン＋被検物投与群およびストレプトゾトシン投与群は投与経過期間が延びるほど回避反応スコアが高くなり、約１４日で回避反応スコアは一定となった。これにより、ストレプトゾトシンの投与によって、糖尿病性末梢神経障害を発症したと考えられた。 回避 In the control group, the avoidance response score was 1 or less stably. On the other hand, in the streptozotocin + test substance administration group and the streptozotocin administration group, the avoidance response score became higher as the administration elapsed time became longer, and the avoidance response score became constant at about 14 days. Thus, it was considered that the administration of streptozotocin caused diabetic peripheral neuropathy.
　２１日目に被検物の投与を開始し（図４中では、「ＬＥＭ投与」と示した。）、この日よりストレプトゾトシン＋被検物投与群は、回避反応スコアが減少した。 投 与 Administration of the test substance was started on day 21 (indicated as “LEM administration” in FIG. 4), and from that day on, the avoidance response score of the group treated with streptozotocin and the test substance decreased.
　以上のことから、本発明に係る糖尿病性末梢神経障害の予防又は治療用組成物は、糖尿病性末梢神経障害を発症した後であっても、末梢神経障害が改善され、治療効果があることを示している。 From the above, the composition for preventing or treating diabetic peripheral neuropathy according to the present invention, even after the onset of diabetic peripheral neuropathy, shows that peripheral neuropathy is improved and has a therapeutic effect. Is shown.
　本発明に係る組成物は、糖尿病によって誘発される末梢神経障害の予防又は治療に利用することができる。

  The composition according to the present invention can be used for preventing or treating peripheral neuropathy induced by diabetes.

Claims (5)

  1. 　シイタケ菌糸体の抽出物を含有することを特徴とする、糖尿病性末梢神経障害の予防又は治療用組成物。 (4) A composition for preventing or treating diabetic peripheral neuropathy, comprising an extract of Shiitake mushroom mycelium.
  2. 　シイタケ菌糸体の抽出物を含有することを特徴とする、糖尿病性末梢神経障害の予防又は治療用医薬組成物。 A pharmaceutical composition for preventing or treating diabetic peripheral neuropathy, comprising an extract of Shiitake mushroom mycelium.
  3. 　前記シイタケ菌糸体の抽出物の１日当たりの投与量が、乾燥粉末重量として４ｍｇ／ｋｇ体重以上であることを特徴とする請求項２に記載された糖尿病性末梢神経障害の予防又は治療用医薬組成物。 The pharmaceutical composition for preventing or treating diabetic peripheral neuropathy according to claim 2, wherein a daily dose of the extract of Shiitake mycelium is 4 mg / kg body weight or more as a dry powder weight. object.
  4. 　シイタケ菌糸体の抽出物を含有し、糖尿病性末梢神経障害の予防又は治療効果を有する加工食品。 (4) A processed food containing an extract of Shiitake mushroom mycelium and having an effect of preventing or treating diabetic peripheral neuropathy.
  5. 　請求項２に記載された糖尿病性末梢神経障害の予防又は治療用医薬組成物を対象に投与することを含む、糖尿病性末梢神経障害を予防又は治療する方法（人を手術・治療・診断する方法を除く）。

          A method for preventing or treating diabetic peripheral neuropathy (a method for operating, treating, or diagnosing a human being), comprising administering to a subject the pharmaceutical composition for preventing or treating diabetic peripheral neuropathy according to claim 2. except for).

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