JP6454592B2 - Acetylcholinesterase inhibitor - Google Patents

Description

  The present invention relates to an inhibitor having an acetylcholinesterase inhibitory action.

  Acetylcholine, a neurotransmitter, plays an important role by binding to receptors on target cells or cholinergic neurons.

  This acetylcholine is synthesized by the synthase choline acetyltransferase and is kept at a constant concentration in the body by being degraded by the degrading enzyme acetylcholinesterase. That is, the concentration of acetylcholine in the body is controlled by the balance between acetylcholine synthase and degrading enzymes. When the degradation of acetylcholine by acetylcholinesterase is promoted, the amount of acetylcholine in the neurotransmitter decreases. Decreasing acetylcholine concentration weakens cholinergic nerve transmission. Insufficient cholinergic nerve transmission is considered to be one of the causes of decreased learning function and the onset of Alzheimer's dementia.

  The effects of non-neuronal acetylcholine on the skin have not yet been fully elucidated, but acetylcholine present on the skin surface has been reported to contribute to maintaining a healthy skin state and promoting the repair of skin damage. It was. The expression level of acetylcholinesterase in the epidermal cells exposed to ultraviolet radiation increased, and the matrix metalloprotease also increased. Addition of acetylcholine or acetylcholinesterase inhibitor decreased the expression level of matrix metalloprotease. Therefore, a skin damage repair effect is expected through inhibition of acetylcholinesterase activity induced by ultraviolet damage (Non-patent Document 1).

  In recent years, it has been reported that Alzheimer's dementia and its peripheral symptoms can be improved or the progression of the disease state can be delayed by inhibiting the acetylcholinesterase activity and increasing the concentration of acetylcholine, which is clinically used. ing.

  As the acetylcholinesterase inhibitor used as a therapeutic drug for Alzheimer, donepezil, galantamine, rivastigmine and the like are known (Non-patent Document 2).

  However, these acetylcholinesterase inhibitors have problems in that they have gastrointestinal side effects such as vomiting, nausea and diarrhea, and dry skin.

  Therefore, the present inventor targeted the above-mentioned acetylcholinesterase, not only for Alzheimer's dementia, but also for the therapeutic and / or prophylactic effect on psychiatric anxiety and depressive symptoms due to a decrease in learning function due to acetylcholine reduction, or acetylcholinesterase activity. In order to obtain substances with no or little side effects, which have a preventive effect on skin problems due to an increase, a new material having an acetylcholinesterase inhibitory action was searched.

Suenobu Noriko, Saito Marie: Inhibition of epidermal acetylcholinesterase activity by essential oil and effect on skin, FRAGRANCE JOURNAL 4, 20, 24-24, 2014. Yu Nakamura: About the Alzheimer type treatment for dementia, Journal of Senile Dementia, 19 (7), 104-108

  The present invention has been made in view of such circumstances, and is an acetylcholinesterase inhibitor having high safety and excellent acetylcholinesterase inhibitory activity, and an effective pharmaceutical composition, cosmetic, bathing agent or The object is to provide a food composition.

The present invention is an acetylcholinesterase inhibitor comprising, as an active ingredient, an extract derived from leaves of the genus Rubiaceae.
The present invention also provides a therapeutic / preventive agent for Alzheimer-type senile dementia characterized by containing an extract derived from the leaves of the genus Rubiaceae, and also for treating / preventing skin troubles caused by increased acetylcholinesterase activity. It also provides an agent.

  According to the present invention, it is possible to provide an acetylcholinesterase inhibitor which is excellent in safety and exhibits a high inhibitory action on acetylcholinesterase activity and a composition relating to the use thereof.

  The present inventors conducted an experiment on the inhibitory action of acetylcholinesterase on an extract of the leaves of the genus Rubiaceae (hereinafter referred to as butterfly (Chitoha)), whereby the extract of butterfly has a strong acetyl content. The present invention has been completed by clarifying that it has a cholinesterase inhibitory action.

  Rubiaceae's Uncaria plant is a species of Uncaria rhynchophylla (Miq.) Miq. Or it is chosen from 2 or more types.

  The above butterfly is a leaf of the genus Kasakazura from China and Japan. In the present invention, only leaves are used. The leaves of the genus Capillaris used in the acetylcholinesterase agent of the present invention are discarded in large quantities at the time of harvest in the production area and can be easily obtained as a raw material. Furthermore, the leaf has a lower water content than the roots and stems and is easy to dry artificially, so it does not require special equipment and can be handled easily. The use of only leaves is preferable not only because it can be obtained in a large amount and at a lower cost than when other roots and stems are used, but also because it is easy to process food.

  A method for extracting an extract of butterfly that can be used as an acetylcholinesterase inhibitor is not particularly limited. For example, butterfly can be extracted using an extraction solvent of water or a water-soluble solvent. Moreover, it can also hydrolyze after extracting using an extraction solvent. Moreover, after decomposing | disassembling using an enzyme, it can also extract using an extraction solvent.

  Examples of the extraction solvent include water, physiological saline, alcohols (for example, lower alcohols such as methanol, absolute ethanol and ethanol, polyhydric alcohols such as propylene glycol and 1,3-butylene glycol), ketones such as acetone, and ethyl. Examples include ethers such as ether, dioxane, acetonitrile, and ethyl acetate, xylene, benzene, and chloroform. Among these, it is preferable to use water. These may be used alone or in combination of two or more.

  After performing the above extraction, the target extract can be obtained by removing solids by filtration, centrifugation, decantation, or the like.

  The above extract of butterfly does not lose its activity even if the pH is adjusted as necessary and the polymer is removed by gel filtration or ultrafiltration. In addition, the enzyme degradation product or a fractional component thereof may be concentrated by a method such as vacuum concentration, ultrafiltration or freeze concentration, or may be dry powdered by a method such as freeze drying, spray drying or flat plate drying. it can.

  The acetylcholinesterase inhibitor of the present invention is expected to be effective in the treatment and / or prevention of Alzheimer-type senile dementia, decreased learning function, depression, and mental anxiety. The present invention also provides a composition capable of preventing and improving skin troubles caused by acetylcholinesterase.

  An acetylcholinesterase inhibitor blended with butterfly can be used as a blending component in, for example, pharmaceuticals, health foods, drinks, skin cosmetics, skin patch sheets, skin external preparations, bath preparations and the like. The external preparation for skin includes quasi-drugs, designated quasi-drugs, external medicines, and the like, and the dosage form can be arbitrarily selected according to the purpose.

  It does not specifically limit as a dosage form of this invention, Either oral administration or parenteral administration may be sufficient. Specifically, an extract, a capsule, a granule, a pill, a powder, an injection, a decoction / soaking agent, a liquid, a drink and the like can be mentioned. These preparations can be produced by a conventional method in the pharmaceutical technical field, and can be produced, for example, by a method described in the Japanese Pharmacopoeia. These preparations can be safely administered to mammals including humans.

(Preparation method of butterfly extract)
Ten times the amount of water was added to 500 g of butterfly powder, and extracted by heating at 100 ° C. for 30 minutes. The extract was centrifuged and concentrated. The concentrated solution was dried by spray drying to obtain 165.7 g of a butterfly extract.

(Test example) [Measurement of acetylcholinesterase inhibitory activity]
The acetylcholinesterase inhibitory activity was measured using MATP +, which is a substrate having high selectivity for acetylcholinesterase. That is, using a 96-well microplate, 50 μL of various concentrations of butterfly extract dissolved in phosphate buffer (pH 7.4), 50 μL of acetylcholinesterase enzyme solution, 50 μL of substrate MATP + solution, and coloring reagent DNTB (5,5- After adding 50 μL of dithiobis (2-nitrobenzoic acid) solution and mixing, the mixture was reacted at 37 ° C. for 30 minutes, and the absorbance at 415 nm was measured to determine the inhibition rate of butterfly acetylcholinesterase. In the blank, 50 μL of phosphate buffer (pH 7.4) was added instead of the enzyme solution. The acetylcholinesterase inhibition rate was calculated according to Equation 1.
(Formula 1)
Acetylcholinesterase inhibition rate (%) = {1− (A−B) / A} × 100

A = (Absorbance after enzyme reaction without addition of test sample-absorbance of the blank)
B = (absorbance after enzyme reaction of each concentration of test sample-absorbance of each blank)

  As shown in Table 1, the butterfly extract (1250 μg / ml) exhibited an inhibitory effect equivalent to that of the positive control (conventional product) on the acetylcholinesterase activity.

  Examples of pharmaceutical compositions, food and beverage compositions, cosmetics, and the like containing the acetylcholinesterase inhibitory action of the butterfly extract of the present invention are shown below. The composition is expressed in mass%.

(Example 1) Soft capsule wt%
Butterfly Extract 1.0
Vitamin E 20.0
Wheat germ oil 15.0
Glycerin fatty acid ester 5.0
Beeswax 2.0
Soybean oil to 100.0

The above ingredients were mixed and filled into a ceratin film made of gelatin and glycerin to obtain a soft capsule according to a conventional method.

(Example 2) Granules wt%
Butterfly Extract 1.0
Starch 30.0
Lactose 49.0
Crystalline cellulose to 100.0

The above respective parts by weight were uniformly mixed and prepared by a conventional method to prepare granules.

(Example 3) Tablet wt%
Butterfly Extract 1.0
Crystalline cellulose 1.5
Vitamin C 20.0
Fragrance 1.3
Guam gum 0.06
DK ESTER 1.5
Powdered sugar to 100.0

(Example 4) Hard capsule wt%
Butterfly Extract 1.0
Dextrin 24.0
Glycerin fatty acid ester 1.0
Powdered sugar to 100.0

The above components were mixed and filled into a capsule container made of gelatin to obtain a hard capsule according to a conventional method.

(Example 5) Pack wt%
Butterfly Extract 0.5
Talc 25.0
Propylene glycol 6.0
KCl 0.1
Perfume proper amount kaolin to 100.0

(Example 6) Nutritional cream wt%
Butterflyfish extract 0.1
Stearic acid 2.5
Stearyl alcohol 8.0
Squalane 5.0
Octyldodecanol 7.0
Glyceryl monostearate 4.0
Preservative Appropriate amount of perfume Appropriate amount of glycerin 6.0
Sorbitol 13.0
Potassium lactate 0.5
Purified water to 100.0

(Example 7) Latex wt%
Butterflyfish extract 0.1
Stearic acid 2.0
Cetanol 2.0
Vaseline 2.0
Lanolin alcohol 1.5
Liquid paraffin 9.0
Squalane 3.5
Sorbitol 10.0
Triethanolamine 1.0
Propylene glycol 5.0
Preservative appropriate amount perfume appropriate amount purified water to 100.0

  The production methods of Examples 5 to 7 were in accordance with ordinary cosmetic methods.

(Example 8) Bath agent composition wt%
Butterfly leaf extract 0.5
Sodium bicarbonate 10.0
Sodium chloride 5.0
Dye 0.2
Fragrance 0.95
1,3-butylene glycol 0.02
Monooleic acid polyoxyethylene sorbitan (20.E.0) 0.05
Silicic anhydride 0.5
Anhydrous sodium sulfate to 100.0

  The production method of Example 8 was according to the method of a normal bath agent composition.

  From this result, butterfly extract has an excellent acetylcholinesterase inhibitory action, Alzheimer type senile dementia caused by acetylcholinesterase, decreased learning function, depression, mental anxiety or UV photo-aging, It can be expected that skin troubles can be delayed. As a result, the composition containing the butterfly extract has an inhibitory action on acetylcholinesterase, Alzheimer's dementia caused by acetylcholinesterase, decreased learning function, depression, mental anxiety or UV photo-aging, skin aging It is considered to have prevention and improvement of trouble.

Pharmaceuticals, cosmetics and food compositions containing an extract of butterfly that has been found to have an acetylcholinesterase inhibitory action according to the present invention are not only for Alzheimer's dementia, but also for the treatment of reduced learning function, depression, mental anxiety and / or It is thought that it can contribute to prevention. In addition, a preventive effect can be expected for skin troubles caused by acetylcholinesterase.

Claims (2)

An acetylcholinesterase inhibitor comprising as an active ingredient an extract derived from the leaves of the genus Rubiaceae. A therapeutic / preventive agent for Alzheimer-type senile dementia, comprising as an active ingredient an extract derived from leaves of the genus Rubiaceae.