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WO2017006407A1 - Inhibiteur de la mort des cellules nerveuses, agent anti-maladie d'alzheimer, inhibiteur de l'hypofonction cérébrale, médicament ou aliment présentant un effet anti-maladie d'alzheimer ou un effet d'inhibition de l'hypofonction cérébrale, et méthode de production d'inhibiteur de la mort des cellules nerveuses - Google Patents

Inhibiteur de la mort des cellules nerveuses, agent anti-maladie d'alzheimer, inhibiteur de l'hypofonction cérébrale, médicament ou aliment présentant un effet anti-maladie d'alzheimer ou un effet d'inhibition de l'hypofonction cérébrale, et méthode de production d'inhibiteur de la mort des cellules nerveuses Download PDF

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Publication number
WO2017006407A1
WO2017006407A1 PCT/JP2015/069363 JP2015069363W WO2017006407A1 WO 2017006407 A1 WO2017006407 A1 WO 2017006407A1 JP 2015069363 W JP2015069363 W JP 2015069363W WO 2017006407 A1 WO2017006407 A1 WO 2017006407A1
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Prior art keywords
cell death
liquid
methanol
distributed
death inhibitor
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English (en)
Japanese (ja)
Inventor
高橋　知也
一紅 潘
淑芳 ▲温▼
亦晃 呂
凱安 ▲荘▼
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AOB KEIOH GROUP CORP
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AOB KEIOH GROUP CORP
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Priority to JP2017526812A priority Critical patent/JP6365914B2/ja
Priority to PCT/JP2015/069363 priority patent/WO2017006407A1/fr
Priority to CN201580081025.1A priority patent/CN107708717B/zh
Priority to TW105118524A priority patent/TWI610676B/zh
Publication of WO2017006407A1 publication Critical patent/WO2017006407A1/fr
Anticipated expiration legal-status Critical
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/21—Esters, e.g. nitroglycerine, selenocyanates
    • A61K31/215—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids
    • A61K31/22—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids of acyclic acids, e.g. pravastatin
    • A61K31/222—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids of acyclic acids, e.g. pravastatin with compounds having aromatic groups, e.g. dipivefrine, ibopamine
    • 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
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/19—Acanthaceae (Acanthus family)

Definitions

  • the present invention relates to a nerve cell death inhibitor, an anti-Alzheimer's disease agent, an anti-brain function-lowering agent, a drug or food having an anti-Alzheimer's disease action or an anti-brain function-lowering action, and a method for producing a nerve cell death inhibitor.
  • Alzheimer's disease Alzheimer's disease
  • Lewy body dementia Lewy body dementia
  • cerebrovascular dementia cerebrovascular dementia
  • amyloid cascade hypothesis is widely supported as the mechanism of Alzheimer's disease.
  • the amyloid cascade hypothesis is that amyloid ⁇ , an amyloid protein, accumulates in the brain, resulting in Alzheimer's neurofibrillary tangles, neuronal death or loss due to direct or indirect toxicity, etc. It is a hypothesis that various symptoms of the disease occur.
  • the hypothesis is widely supported at present, and serves as a guideline for research and development of preventive and therapeutic methods (see, for example, Non-Patent Document 1).
  • the white crane reishi (Rhinacanthus natusus (L.) Kurz. Also called white crane reishi grass) is an evergreen small shrub belonging to the genus Linacanthus vulgaris, which originates in the Deccan Plateau in southern India.
  • Hakutsuru Ganoderma is known to have an antibacterial action against anthelmintic, anti-inflammatory, and skin fungi (see, for example, Non-Patent Document 3), mainly in China, Taiwan, etc., and recently in Japan, Used as food.
  • the white crane reishi has an active oxygen scavenging ability (for example, see Patent Document 1), has an excretion promoting action (for example, see Patent Document 2), and antiallergic. It is disclosed that there is an action (for example, see Patent Document 3) and that it has an antitumor action (for example, see Patent Document 4).
  • linacantin C contained in a predetermined distribution, a predetermined fraction, or a white crane reishi obtained from a white crane reishi extract by a predetermined method has a neuronal cell death inhibitory effect on the toxicity of amyloid ⁇ . unknown.
  • the present invention provides a neuronal cell death inhibitor, an anti-Alzheimer's disease agent, an anti-brain function-lowering agent, and an anti-Alzheimer's disease action or an anti-brain function-lowering drug or food having an excellent neuronal death inhibitory effect.
  • Another object of the present invention is to provide a method for producing a nerve cell death inhibitor capable of producing a nerve cell death inhibitor having an excellent nerve cell death inhibitory action.
  • Ethanol extract of Hakutsuru Reishi is liquid-liquid distributed with hexane and methanol, and the liquid distributed to the methanol is liquid-liquid distributed with methylene dichloride and water and distributed to the methylene dichloride.
  • the partition is subjected to silica gel column chromatography using an elution solvent composed of n-hexane, ethyl acetate and anhydrous methanol, The nerve cell death inhibitor which contains only the predetermined fraction which is a fraction eluted with the said elution solvent from a silica gel column as an active ingredient.
  • the predetermined fraction is eluted in a volume of 0.1 to 4 times the volume of silica gel used in the silica gel column chromatography.
  • a neuronal cell death inhibitor which is a fraction eluted when a solvent is used.
  • a neuronal cell death inhibitor containing only linacantin C as an active ingredient [6] A neuronal cell death inhibitor containing only linacantin C as an active ingredient.
  • An anti-Alzheimer's disease agent comprising the neuronal cell death inhibitor according to any one of [1] to [6] as an active ingredient.
  • a method for producing a nerve cell death inhibitor comprising only the predetermined distribution as an active ingredient.
  • a method for producing a neuronal cell death inhibitor comprising only the predetermined fraction as an active ingredient, comprising the fourth step obtained in this order.
  • a neuronal death inhibitor an anti-Alzheimer's disease agent, an anti-brain function-lowering agent, an anti-Alzheimer's disease action or an anti-brain function having an excellent inhibitory effect on neuronal cell death.
  • a pharmaceutical or food having a lowering action can be provided.
  • the above [1] is, for example, “use of the predetermined distribution as an active ingredient for producing a neuronal death inhibitor containing only the following predetermined distribution as an active ingredient.
  • the ethanol extract of Hakutsuru Ganoderma is liquid-liquid distributed with hexane and methanol, and the partition distributed to methanol is liquid-liquid distributed with methylene dichloride and water, the distribution is distributed to the methylene dichloride. It can also be expressed as “the predetermined distribution which is an object”.
  • the predetermined fraction as an active ingredient for producing a nerve cell death inhibitor containing only the following predetermined fraction as an active ingredient
  • the Hakutsuru Reishi lawn ethanol extract was liquid-liquid distributed with hexane and methanol, and the methanol-distributed distribution was liquid-liquid distributed with methylene dichloride and water and distributed to the methylene dichloride.
  • the predetermined fraction which is a fraction eluted from a silica gel column with the elution solvent when the partition is subjected to silica gel column chromatography using an elution solvent composed of n-hexane, ethyl acetate and anhydrous methanol. "Can also be expressed.
  • the above [6] may be expressed as, for example, “use of linacantin C as an active ingredient for producing a neuronal cell death inhibitor containing only linacantin C as an active ingredient”. it can.
  • [7] above for example, “use of the neuronal cell death inhibitor according to any of [1] to [6] as an active ingredient for producing an anti-Alzheimer's disease agent”. It can also be expressed as follows.
  • [8] for example, “use of the nerve cell death inhibitor according to any of [1] to [6] as an active ingredient for producing an antibrain function-lowering agent”.
  • the anti-Alzheimer's disease agent according to [7] or the above [8] for producing a pharmaceutical or food having an anti-Alzheimer's disease action or an anti-brain function lowering action "Use of the described antibrain function-lowering agent as an active ingredient.”
  • the nerve cell death inhibitor the anti-Alzheimer's disease agent, the anti-brain function-lowering agent, the drug or food having the anti-Alzheimer's disease action or the anti-brain function-lowering action, and the method for producing the nerve cell death inhibitor of the present invention will be described.
  • the neuronal cell death inhibitor containing only a predetermined distribution which is a distribution as an active ingredient is a "first step of obtaining an ethanol extract by extracting white crane reishi with ethanol, Liquid-liquid partition with hexane and methanol to obtain a partition distributed to the methanol, and liquid-liquid partition of the partition distributed to methanol with methylene dichloride and water to the methylene dichloride. And a third step of obtaining a predetermined distribution which is a distribution to be distributed in this order, and a method for producing a neuronal cell death inhibitor containing only the predetermined distribution as an active ingredient ” It can be.
  • the predetermined distribution in the present invention can be obtained under various conditions as long as the conditions described are satisfied.
  • the “partition” means a purified product obtained by liquid-liquid partition.
  • “containing only a predetermined distribution as an active ingredient” refers to containing only a predetermined distribution as an active ingredient having an inhibitory action on nerve cell death, and is not an active ingredient ( It does not deny that it contains various solvents and auxiliary additives.
  • the raw Hakutsuru Reishi leaves, stems, roots, flowers, etc. are collected at an appropriate time and then used as they are or are usually subjected to a drying process such as ventilation drying to extract raw materials. Can be used.
  • the white crane reishi as an extraction raw material is preferably used after being crushed or chopped.
  • ethanol simply means a “solvent containing ethanol as a main component (containing 50% or more of ethanol)”. That is, “ethanol” includes not only so-called anhydrous ethanol but also a mixed solvent of ethanol and another solvent.
  • a mixed solvent of ethanol and water hydroous ethanol
  • the extraction temperature is usually 0 to 100 ° C., preferably 5 to 50 ° C.
  • the extraction time is about 1 hour to 10 days, and the amount of solvent is usually 1 to 30 times by weight, preferably 5 to 10 times by weight per dry raw material.
  • the extraction operation may be stirring or simple immersion.
  • the extraction operation may be repeated a plurality of times as necessary. Moreover, you may combine extraction operation from which conditions differ. Furthermore, it is preferable to perform an operation of removing insoluble residues from the crude extract obtained by the above operation by filtration or centrifugation.
  • Liquid-liquid distribution can also be carried out by known methods usually used industrially and experimentally.
  • Liquid-liquid partitioning is a method called two-phase partitioning, two-phase solvent partitioning, liquid-liquid extraction, etc., and is a method of recovering a target compound by using a difference in solubility using two kinds of solvents having different polarities. .
  • After the liquid-liquid partition it is preferable to obtain a partition in a dry state by removing the solvent. However, this is not the case when the removal of the solvent is unnecessary for the purpose of use.
  • methanol simply means “a solvent containing methanol as a main component (containing 50% or more of methanol)”. That is, “methanol” includes not only so-called anhydrous methanol but also a mixed solvent of methanol and another solvent. When performing liquid-liquid partitioning, it is preferable to use a mixed solvent of methanol and water, particularly 90% methanol.
  • anhydrous methanol refers to anhydrous methanol and does not include a mixed solvent containing methanol. It can be obtained by a “method for producing a nerve cell death inhibitor containing only a predetermined distribution as an active ingredient”.
  • the ethanol extract of Hakutsuru Reishi is liquid-liquid distributed with hexane and methanol
  • the liquid distributed to methanol is liquid-liquid distributed with methylene dichloride and water and distributed to methylene dichloride.
  • Neuronal cell death inhibitor contained as an active ingredient includes “first step of extracting white crane reishi with ethanol to obtain an ethanol extract, and liquid-liquid distribution of the ethanol extract with hexane and methanol to the methanol. A second step of obtaining a partition to be distributed; and a liquid-liquid partition of the partition distributed to methanol with methylene dichloride and water, The third step to obtain a partition distributed to methylene chloride and the partition distributed to methylene dichloride were subjected to silica gel column chromatography using an elution solvent composed of n-hexane, ethyl acetate and anhydrous methanol. And a fourth step of obtaining a predetermined fraction which is a fraction eluted from the silica gel column with the elution solvent in this order, and a neuronal cell death inhibitor containing the predetermined fraction as an active ingredient.
  • the predetermined fraction in the present invention can be obtained under various conditions as long as the described conditions are satisfied.
  • the “fraction” means a purified product obtained by fractionation by column chromatography.
  • “containing only a predetermined fraction as an active ingredient” means containing only a predetermined fraction as an active ingredient having a nerve cell death inhibitory action, and is not an active ingredient. It does not deny that it contains ingredients (various solvents, auxiliary additives, etc.). Since the extraction and the liquid-liquid distribution have already been described, the description thereof will be omitted. After silica gel column chromatography, it is preferable to obtain a fraction in a dry state by removing the elution solvent. However, this is not the case when the removal of the solvent is unnecessary for the purpose of use.
  • the predetermined fraction is more preferably a fraction that elutes when an elution solvent having a volume of 0.1 to 4 times the volume of silica gel used for silica gel column chromatography is used. It is even more preferable that it is a fraction that elutes when an elution solvent having a volume of 0.2 to 2.5 times is used (see test examples described later).
  • the predetermined distribution and the predetermined fraction may be obtained by further performing a purification method other than the above as long as the object of the present invention is not impaired.
  • the solvent and the mixed solvent used for obtaining the predetermined distribution and the predetermined fraction can be replaced with a solvent or a mixed solvent having the same degree of polarity.
  • a solvent it can be used to obtain a predetermined distribution and a predetermined fraction.
  • solvents include water, various alcohols, esters such as ethyl acetate, linear, branched and cyclic hydrocarbons, methylene dichloride (dichloromethane), chloroform, acetone, various ethers, etc.
  • those suitable for the method and purpose can be used.
  • the predetermined distribution and the predetermined fraction in the present invention contain linacantin C as one component.
  • the linacantin C in the “nerve cell death inhibitor containing only linacantin C as an active ingredient” of the present invention may be obtained by extracting and purifying white crane reishi as a raw material (see experimental examples described later). It may be obtained from other plants. Also, linacantin C may be obtained by synthesis. Linacantin C is a compound represented by the following chemical formula (1).
  • the linacantin C of the present invention may be linacantin C in the mechanism of action in the body, and may be in the form of a pharmaceutically acceptable salt before administration.
  • the predetermined distribution, the predetermined fraction, and linacantin C have the effect of suppressing neuronal cell death due to the toxicity of amyloid ⁇ , as shown in the experimental examples described later. For this reason, it is thought that the nerve cell death inhibitor of this invention which uses these as an active ingredient has the effect
  • the “anti-Alzheimer's disease agent” in the present specification can be used for at least one of prevention and treatment of Alzheimer's disease.
  • the “anti-brain function-lowering agent” can be used for at least one of prevention and treatment of a decrease in brain function.
  • the route of administration of the neuronal death inhibitor, anti-brain function-lowering agent, anti-Alzheimer's disease agent and pharmaceuticals (herein, drugs for internal use or internal use) of the present invention is not particularly limited, and examples thereof include oral administration and rectal administration. And mucosal administration such as nasal administration, injection administration such as intravenous administration and subcutaneous administration, and the like.
  • Any dosage form can take the form of a formulation suitable for the administration method. For example, tablets, powders, fine granules, granules, capsules, powders, pills, lozenges and other solid agents, solutions , Suspensions, emulsions, syrups, liquids such as injections, gel preparations, and the like.
  • the neuronal cell death inhibitor, anti-brain function-lowering agent, anti-Alzheimer's disease agent and pharmaceutical of the present invention may be administered as they are, or may be administered together with a pharmacologically acceptable excipient.
  • a pharmacologically acceptable excipient any monosaccharides, disaccharides, polysaccharides, inorganic salts, oils and fats, distilled water, etc. that can be generally used as preparations can be used.
  • additives such as binders, lubricants, dispersants, suspending agents, emulsifiers, diluents, buffers, antioxidants, and bacterial inhibitors can be used.
  • the effective dosage of the neuronal cell death inhibitor, anti-brain function-lowering agent, anti-Alzheimer's disease agent and pharmaceutical of the present invention varies depending on the administration route, dosage form, disease symptom, age of the subject, etc.
  • the predetermined fraction or linacantin C is considered to be usually 0.1 to 1000 mg, preferably 0.5 to 300 mg, more preferably 1 to 100 mg per day for an adult.
  • the content of a predetermined distribution, a predetermined fraction or linacantin C is determined based on the data on the dosage form, the effective dosage, and the dosage as the dosage, and the optimal active ingredient content in the dosage form for each dosage form. Can be set.
  • the pharmaceutical product of the present invention may be an external pharmaceutical product.
  • the form of the external medicine is not particularly limited.
  • an ointment, a cream, a foaming agent, a tape agent, an external preparation etc. can be mentioned, for example.
  • the external medicine of the present invention can contain various pharmaceutical ingredients in a predetermined distribution, a predetermined fraction or linacantin C as required.
  • additives such as a binder, a dispersant, a suspending agent, an emulsifier, a diluent, a buffer, an antioxidant, and a bacteria inhibitor can also be used.
  • Examples of the food of the present invention include foods in the form of tea with the addition of active ingredients and foods in the form of processed foods containing the active ingredients.
  • tea it is preferable to use it by mixing with a dried white crane reishi leaf / stem or root or by mixing with other tea ingredients.
  • tea materials green tea, oolong tea, puer tea, black tea, roasted green tea, brown rice tea, Tochu tea, kashiwanoha tea, mulberry leaf tea, etc., as long as they are used as normal teas, any tea can be used. Can do.
  • the dried white crane reishi leaf / stem or root can be roasted and used in the same manner as other tea ingredients.
  • a form of food containing the active ingredient drinks, jelly, biscuits, tablets, pills, soft capsules, hard capsules, powders, fine granules, granules, etc. Either form can be used.
  • additives such as excipients, binders, lubricants, dispersants, suspending agents, emulsifiers, diluents, buffers, antioxidants, and bacterial inhibitors can be used as food auxiliary materials. .
  • the effective intake amount of the food of the present invention varies depending on the intake form, the health condition of the subject, the age of the subject, etc., but for a given distribution, a given fraction or linacantin C, it is usually 0. It is considered to be 1 to 1000 mg, preferably 0.5 to 300 mg, more preferably 1 to 100 mg.
  • the content of the predetermined distribution, the predetermined fraction or linacantin C in the food of the present invention varies depending on the form of the food, but is usually 0.0001 to 1 wt%, preferably 0.001 to 0.5 wt%. More preferably, it is considered to be 0.01 to 0.1 wt%.
  • Example of distribution / fractionation / isolation of the distribution of the present invention, the fraction of the present invention and linacantin C The distribution / fractionation of the distribution of the present invention and the fraction of the present invention follows the flow shown in FIG. I went.
  • 2 kg of dried roots of Rhinacanthus natusus (L.) Kurz was prepared and pulverized with a general-purpose grinder.
  • extraction with 90% ethanol was performed.
  • the extraction was performed by immersing Hakutsuru Reishi in 20 L of 90% ethanol for 3 days and then refluxing the solvent for 1 hour.
  • the extract and the extraction residue were separated by filter paper.
  • the extraction was performed three times on the same raw material, and the solvent was removed from the extract under reduced pressure to obtain 77.82 g of a dry ethanol extract.
  • the remaining 73.82 g ethanol extract was subjected to liquid-liquid partition with hexane and 90% methanol.
  • 500 mL of 90% methanol was added to the ethanol extract, and then 500 mL of hexane was added and shaken for 1 minute.
  • the solvent was separated for about 10 minutes, and a 90% methanol phase was collected.
  • the same operation was further repeated twice, and the obtained 90% methanol phase was combined, and then the solvent was removed under reduced pressure to obtain 54.32 g of a dried product.
  • predetermined distribution A After removing 4 g from the above-mentioned distribution as an analytical sample, the remaining 50.32 g of the entire distribution was subjected to liquid-liquid distribution with methylene dichloride and water. First, 500 mL of water (purified water) was added to the distribution, and then 500 mL of methylene dichloride was added and shaken for 1 minute. The solvent was separated for about 10 minutes and the methylene dichloride phase was collected. Further, the same operation was repeated twice, and the obtained methylene chloride phase was combined, and then the solvent was removed under reduced pressure to obtain 27.55 g of a predetermined distribution. In the following description, the predetermined distribution obtained here is referred to as “predetermined distribution A”.
  • 20.00 g of the predetermined distribution A was subjected to silica gel column chromatography.
  • 20.00 g of a predetermined distribution A and 60 g of silica gel (a general-purpose product of 0.063 to 0.2 mm) were mixed.
  • a column (open column with a diameter of 6 cm, a length of 34 cm, and a volume of about 204 mL) packed with 500 g of silica gel (0.040 to 0.063 mm general-purpose product) was prepared, and an elution solvent (n-hexane: ethyl acetate: anhydrous)
  • a mixture of a predetermined partition A and silica gel was placed, and elution with an elution solvent was performed.
  • fraction 1-2 (solvent amount of about 120 to 480 mL), obtained in the 25th to 39th test tube, fraction 1-3 (solvent amount of about 480 to 780 mL), obtained in the 40th to 70th test tube
  • fraction 1-4 (solvent amount: about 780 to 1400 mL)
  • the product obtained with the remaining elution solvent, acetone and absolute ethanol was designated fraction 1-5.
  • the fractions 1-1 to 1-4 are predetermined fractions.
  • silica gel column chromatography method is different from the method of obtaining the predetermined fraction A, only the portions different from the above method will be described.
  • a predetermined distribution A and 10 g of silica gel (a general-purpose product of 0.063 to 0.2 mm) were mixed.
  • fractions 2-1 to 2-3 were analyzed by comparing 1 HNMR and 13 CNMR data with literature values (Journal of Natural Products, 59, 808-811, 1996). It was confirmed that the fraction 2-2 was linacantin C. The amount of obtained linacantin C was 444.2 mg.
  • a nuclear magnetic resonance spectrum apparatus As a nuclear magnetic resonance spectrum apparatus, a JEOL JNM-GSX500 type nuclear magnetic resonance spectrum apparatus (manufactured by JEOL Ltd.) was used.
  • test example on neuronal cell death inhibitory action of predetermined partition, predetermined fraction and linacantin C In this test example, cytotoxicity by amyloid ⁇ and predetermined partition, predetermined fraction and neuronal cell death of linacantin C A test for inhibitory action was performed. For the evaluation of neurotoxicity of amyloid ⁇ , amyloid ⁇ (25-35), which is known as the action center of neurotoxicity of amyloid ⁇ , was used.
  • PC12 cells rat adrenal pheochromocytoma-derived cell line
  • BCRC Bioresource Collection and Research Center, Bioresource Conservation and Research Center, Taiwan
  • the obtained PC12 cells were prepared using RPMI-1640 medium supplemented with 5% fetal bovine serum, 10% horse serum, 100 units / ml penicillin and 100 mg / ml streptomycin (both obtained from Invitrogen-Gibco), 5% carbonic acid.
  • the culture was performed at 37 ° C. and saturated humidity in the presence of gas.
  • a 96-well microplate coated with poly-L-lysine was used to evaluate the cytotoxicity of amyloid ⁇ (25-35).
  • the microplate 2 ⁇ 10 4 / 100 ⁇ L of PC12 cells per well were seeded in the presence of 5% carbon dioxide, 37 ° C., and cultured under saturated humidity. After 24 hours of culture, 0.1% DMSO (manufactured by JT Baker) is used for the control group, each sample (see below) is used for the evaluation group, and 10 ⁇ M galantamine hydrobromide (anti-Alzheimer's disease agent) is used for the positive control group. It is known that it has a neuroprotective action, obtained through Sigma-Aldrich.
  • test sections were tested for both those with and without the addition of 15 ⁇ M amyloid ⁇ (25-35) (obtained through Sigma-Aldrich) (described later). 72 hours after the start of the culture, the cell viability for each test group was calculated by measuring the absorbance at 570 nm by a predetermined method using MTT (Molecular Probes).
  • cell viability was calculated for the control, the positive control, and each sample without adding amyloid ⁇ .
  • cell viability was calculated for each sample under the same conditions except that amyloid ⁇ was added.
  • the value of the cell viability when amyloid ⁇ is added is divided by the value of the cell viability when amyloid ⁇ is not added, and the value calculated by the calculation is multiplied by 100 to give% units.
  • the cell viability recovery rate was obtained by subtracting the cell viability in the control from there. This cell survival recovery rate is an index showing how much the cell survival rate has recovered from the control in a state where the influence of the sample on the cell survival rate is corrected.
  • Samples include a predetermined distribution A (methylene dichloride distribution), a predetermined fraction 1-1, a predetermined fraction 1-2, a predetermined fraction.
  • Fraction 1-3 which is a predetermined fraction, and linacantin C (fraction 2-2) were used.
  • concentration of each sample the screening regarding cytotoxicity etc. was performed in advance and the appropriate density
  • FIG. 3 (a) is a table showing test results for positive control
  • FIG. 3 (b) is a table showing test results for each sample of the present invention.
  • amyloid ⁇ has high toxicity to PC12 cells (see the control item in FIG. 3).
  • the predetermined distribution A the predetermined fraction (fractions 1-1 to 1-4) and linacantin C showed a high cell survival recovery rate.
  • the cell viability recovery rate of the fraction 1-1 and the fraction 1-2 was comparable to that of galantamine hydrobromide.
  • the cell viability recovery rate by linacantin C is 30.2% while the concentration is as low as 1 ⁇ M. Since the cell survival recovery rate by 10 ⁇ M galantamine hydrobromide is 17.2%, it was possible to find the fact that linacantin C shows a particularly high cell survival recovery rate.
  • the nerve cell death inhibitor of the present invention has a good effect of suppressing nerve cell death due to the toxicity of amyloid ⁇ , that is, an excellent nerve cell death inhibitory effect. For this reason, it is thought that the nerve cell death inhibitor of this invention which uses these as an active ingredient has the effect
  • Example 10 By the following examples, a method for preparing pharmaceuticals and foods containing linacantin C, which is an active ingredient of the neuronal death inhibitor of the present invention and an active ingredient of the anti-Alzheimer's disease agent or anti-brain function-lowering agent of the present invention. Describe. Of course, it is also possible to change the item of linacantin C to a predetermined distribution or a predetermined fraction.
  • Linacantin C a tablet is prepared according to the following formulation.
  • Linacantin C 0.2g Lactose 95.8g 2.0g dried corn starch Talc 1.8g Calcium stearate 0.2g
  • Linacantin C 0.2 g
  • talc 1.8 g
  • calcium stearate 0.2 g
  • a tablet is produced by a conventional method using a single-shot tablet press.
  • hard capsules (360 mg per capsule) are prepared according to the following formulation.
  • Linacantin C (5 g) is mixed with lactose (220 g) and corn starch (110 g), and an aqueous solution of hydroxypropylcellulose (25 g) is added thereto and kneaded.
  • granules are produced by an ordinary method using an extrusion granulator.
  • a hard capsule is prepared by filling the granule into a gelatin hard capsule.
  • soft capsules (170 mg per capsule) are prepared according to the following formulation.
  • Linacantin C 0.5mg Soybean oil 169.5mg (Preparation method)
  • Linacantin C (0.5 g) is added to and mixed with soybean oil (169.5 g).
  • soft capsules are prepared by filling soft capsules using a rotary soybean automatic molding machine according to a conventional method.
  • Pills Using linacantin C pills (100 mg per capsule) are prepared according to the following prescription.
  • Linacantin C 0.5mg Morohaya powder 20.0mg Starch 30.0mg Molasses 20.0mg Tea extract 15.0mg Soy fiber 14.0mg Shellac 0.5mg (Preparation method) After mixing the raw materials with the above blending, adding a suitable amount of water, producing a homogeneous kneaded product with a kneader, rolling the kneaded product, making a round shape using a round machine and drying it to produce a pill To do.
  • a powder (1000 mg per packet) is prepared by the following method using a conventional method.
  • Linacantin C 1mg Lactose 799mg Cornstarch 200mg
  • jelly (100 g) is prepared by a conventional method with the following prescription.
  • Linacantin C 0.002g Gelatin 2.0g Orange juice 20.0g 77.998 g of water (Preparation method)
  • the above ingredients are mixed and heated to 90 ° C. After confirming the dissolution of gelatin, fill the container and cool.
  • a jelly is prepared by solidifying gelatin.
  • an ointment (100 g) is prepared by a conventional method with the following formulation.
  • (Oil phase component) Linacantin C 0.1g White petrolatum 20.0g Mineral oil 20.0g Stearyl alcohol 5.0g Steareth-2 3.0g Propylparaben 0.1g Natural vitamin E 0.1g (Aqueous phase component) 1,3-butylene glycol 5.0 g 0.4 g of phenoxyethanol Polysorbate 60 4.5g Purified water Appropriate amount 100g (Preparation method)
  • the oil phase component and the water phase component are each heated to 80 ° C. to be uniform, and the water phase is added to the oil phase with stirring. After emulsification, the mixture is cooled to prepare an ointment.
  • tape agent (100 g) is prepared by a conventional method using linacantin C according to the following formulation.
  • the pressure-sensitive adhesive solvent and the medicinal component are made uniform, the medicinal component and the transdermal absorption accelerator are added to the pressure-sensitive adhesive solvent, and the composition is stirred at room temperature. This composition is spread on a silicone-treated polyester film, dried at 120 ° C. and cooled, and then the pressure-sensitive adhesive layer is transferred to a polyethylene film to produce a tape agent.

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Abstract

La présente invention concerne un inhibiteur de la mort des cellules nerveuses, un agent anti-maladie d'Alzheimer, un inhibiteur de l'hypofonction cérébrale et un médicament ou un aliment présentant un effet anti-maladie d'Alzheimer ou un effet d'inhibition de l'hypofonction cérébrale, contenant chacun comme principe actif « une partie partitionnée prédéfinie qui est une partie partitionnée dans du dichlorure de méthylène lorsqu'une séparation liquide-liquide est mise en œuvre sur un extrait éthanolique de Rhinacanthus nasuta (L.) Kurz avec de l'hexane et du méthanol et qu'ensuite une séparation liquide-liquide est mise en œuvre sur la partie partitionnée dans le méthanol avec du dichlorure de méthylène et de l'eau », « une fraction prédéfinie qui est une fraction éluée d'une colonne de gel de silice avec un solvant d'élution, ledit solvant d'élution consistant en du n-hexane, de l'acétate d'éthyle et du méthanol anhydre, lorsque la partie partitionnée prédéfinie est soumise à une chromatographie sur colonne de gel de silice à l'aide du solvant d'élution » ou Rhinacanthin C. Selon la présente invention, un inhibiteur de la mort des cellules nerveuses présentant un excellent effet d'inhibition de la mort des cellules nerveuses, un agent anti-maladie d'Alzheimer, un inhibiteur de l'hypofonction cérébrale et un médicament ou un aliment présentant un effet anti-maladie d'Alzheimer ou un effet d'inhibition de l'hypofonction cérébrale peuvent être fournis.
PCT/JP2015/069363 2015-07-04 2015-07-04 Inhibiteur de la mort des cellules nerveuses, agent anti-maladie d'alzheimer, inhibiteur de l'hypofonction cérébrale, médicament ou aliment présentant un effet anti-maladie d'alzheimer ou un effet d'inhibition de l'hypofonction cérébrale, et méthode de production d'inhibiteur de la mort des cellules nerveuses Ceased WO2017006407A1 (fr)

Priority Applications (4)

Application Number Priority Date Filing Date Title
JP2017526812A JP6365914B2 (ja) 2015-07-04 2015-07-04 神経細胞死抑制剤、抗アルツハイマー病剤、抗脳機能低下剤、抗アルツハイマー病作用又は抗脳機能低下作用を有する医薬品又は食品、神経細胞死抑制剤の製造方法、抗アルツハイマー病剤の製造方法、抗脳機能低下剤の製造方法、及び、抗アルツハイマー病作用又は抗脳機能低下作用を有する医薬品又は食品の製造方法
PCT/JP2015/069363 WO2017006407A1 (fr) 2015-07-04 2015-07-04 Inhibiteur de la mort des cellules nerveuses, agent anti-maladie d'alzheimer, inhibiteur de l'hypofonction cérébrale, médicament ou aliment présentant un effet anti-maladie d'alzheimer ou un effet d'inhibition de l'hypofonction cérébrale, et méthode de production d'inhibiteur de la mort des cellules nerveuses
CN201580081025.1A CN107708717B (zh) 2015-07-04 2015-07-04 白鹤灵芝醌c作为神经细胞凋亡抑制剂的应用
TW105118524A TWI610676B (zh) 2015-07-04 2016-06-14 神經細胞死抑制劑、抗阿茲海默症劑、抗腦機能減退劑、具有抗阿茲海默症作用或抗腦機能減退作用的醫藥品及神經細胞死抑制劑的製造方法

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PCT/JP2015/069363 WO2017006407A1 (fr) 2015-07-04 2015-07-04 Inhibiteur de la mort des cellules nerveuses, agent anti-maladie d'alzheimer, inhibiteur de l'hypofonction cérébrale, médicament ou aliment présentant un effet anti-maladie d'alzheimer ou un effet d'inhibition de l'hypofonction cérébrale, et méthode de production d'inhibiteur de la mort des cellules nerveuses

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JP6497791B2 (ja) * 2016-08-09 2019-04-10 株式会社Ａｏｂ慧央グループ ミクログリアｉｌ−６産生抑制剤、中枢神経系抗炎症剤、抗神経変性疾患剤、抗精神神経疾患剤、医薬品及び食品
CN111671089A (zh) * 2020-06-24 2020-09-18 天津市泉又今生物科技有限公司 生物强化细胞营养素在制备抗beta-淀粉样蛋白细胞毒性功能食品中的应用
CN115385795B (zh) * 2022-10-11 2023-11-17 广西中医药大学赛恩斯新医药学院 白鹤灵芝萘醌类单体白鹤灵芝素-c的制备方法及应用

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JP2011190251A (ja) * 2010-02-19 2011-09-29 Arsoa Honsya Corp 抗肥満剤及び脂肪蓄積抑制作用を有する医薬品、食品又は化粧料
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TWI610676B (zh) 2018-01-11
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TW201705970A (zh) 2017-02-16
JP6365914B2 (ja) 2018-08-01
CN107708717B (zh) 2021-05-14

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