JP2013023487A - Anti-glycation agent - Google Patents

Abstract

PROBLEM TO BE SOLVED: To find a substance having an anti-glycation effect from among natural products with high safeness, and to provide an anti-glycation agent containing the substance as an active ingredient.SOLUTION: The anti-glycation agent includes, as an active ingredient, an extract of Osmanthus fragrans var. aurantiacus and/or acteoside. The extract of Osmanthus fragrans var. aurantiacus and/or acteoside preferably has an inhibitory effect on formation of advanced glycation end-products (AGEs) and/or a decomposition promotion effect on AGEs.

Description

  The present invention relates to an anti-glycation agent.

  Carbohydrates are very important as energy sources in living organisms including humans. However, carbohydrates are known to cause glycation reactions with proteins. In the saccharification reaction, the first stage is a non-enzymatic reaction between a carbonyl group of a carbohydrate and an amino group such as a protein, and finally a final glycation product (hereinafter referred to as “AGEs”) from a Schiff base through an Amadori compound. Is a series of reactions to form. Due to the saccharification reaction, the protein is modified non-enzymatically by sugar, and as a result, denaturation of the protein, cross-linking between proteins, and the like occur, resulting in a decrease in protein function.

  The glycation reaction not only causes direct damage by modifying and changing the structure of extracellular matrix constituent proteins such as collagen, but also affects cellular responses by being recognized by receptors with glycated proteins as ligands. Bring. In particular, the effect of the saccharification reaction is serious for diabetic patients whose glucose concentration in the blood is high. It is known that protein glycation contributes to diabetic complications such as diabetic neuropathy, diabetic retinopathy, and diabetic nephropathy. In addition, it is known that the glycation reaction in the blood vessel wall leads to the progression of arteriosclerosis due to the damage of endothelial cells and the accumulation of denatured proteins.

  Further, extracellular matrix components such as collagen occupy a majority of the dry weight in tissues such as bone and skin. Therefore, for example, when collagen is glycated and abnormally crosslinked, osteoporosis, osteoarthritis, etc. occur in bone and cartilage tissue, and elasticity in skin, dullness due to yellowing, etc. occur. . Furthermore, since abnormally cross-linked collagen or the like is less susceptible to degradation by collagenase or the like, the expression of collagenase or the like is induced, causing problems such as degradation to normal collagen.

  Therefore, if the glycation reaction can be suppressed in some way, that is, the formation of AGEs can be suppressed, or the degradation of AGEs can be promoted, the above-mentioned diseases, that is, diabetic complications, arteriosclerosis, osteoporosis, deformability It is expected to be useful for the prevention or treatment of arthropathy. Furthermore, it is expected to be effective in preventing or improving skin elasticity and dullness.

  Conventionally, N-phenacyl thiazolium bromide is known as a compound having an anti-glycation action (Non-patent Document 1). However, this compound has a safety problem and is not suitable as a pharmaceutical or a topical skin preparation. Moreover, as an ingredient derived from a natural product having an anti-glycation action, for example, an extract from mugwort is known (Patent Document 1). However, it is difficult to provide a substance having an anti-glycation action, and there is a strong demand for the development and provision of further new anti-glycation action substances.

  Conventionally, the extracts and acteosides from Buttercups have a matrix metalloproteinase-1 inhibitory action, a cyclic AMP phosphodiesterase inhibitory action, etc., and are known to be useful for the prevention and improvement of skin aging symptoms and obesity. However, it has not been known that an extract or acteoside from Buttercups has an anti-glycation effect (see Patent Document 2).

JP 2001-122758 A JP 2009-67749 A

Vasan S. et al., Nature, July 18, 1996, Vol. 82, No. 6588, p.275-278

  It is an object of the present invention to provide an anti-glycation agent having an anti-glycation action among highly safe natural products and using it as an active ingredient.

  In order to solve the above-mentioned problems, the anti-glycation agent of the present invention is characterized by containing an extract from cinnamon and / or acteoside as an active ingredient. It is preferable that the extract from the cinnamon moss and / or the acteoside has an action of suppressing the formation of final glycation products (AGEs) and / or an action of promoting degradation.

  According to the present invention, it is possible to provide an anti-glycation agent having an excellent anti-glycation effect and high safety.

Embodiments of the present invention will be described below.
The anti-glycation agent of the present embodiment contains an extract from Buttercup and / or Acteoside as an active ingredient.

  Here, in the present embodiment, the “extract from Snapper” is an extract obtained using Snapper as an extraction material, a diluted or concentrated solution of the extract, a dried product obtained by drying the extract, or Any of these crudely purified products or purified products is included.

  Acteoside is a polyphenol having a chemical structure represented by the following formula (I).

  Acteoside can be produced by isolation and purification from a plant extract containing acteoside. Such a plant extract containing acteoside can be obtained by a method generally used for plant extraction. Examples of the plant containing acteoside include, for example, ummingei (scientific name: Osmanthus fragrans var. Aurantiacus).

  Buttercup (Osmanthus fragrans var. Aurantiacus) is an evergreen small tree belonging to the genus Moxae, also known as Katsura, which is native to southern China and can be easily obtained from such areas. Examples of the constituent parts of the cypress that can be used as the raw material for extraction include the aerial parts such as leaves, branches, bark parts, trunk parts, stem parts, fruit parts, flower parts, root parts, or a mixture of these parts. However, it is preferably a flower part.

  Here, the "flower" generally refers to the whole organ involved in sexual reproduction of a seed plant, and is composed of a flower leaf that is a leaf deformation and a flower axis that is a stem deformation. This includes organs such as pupae, petals, stamens, and heart skin. The “floral part” used as an extraction raw material in the present invention includes all of the organs involved in sexual reproduction of seed plants, as well as parts thereof, such as flower leaves, flower coats (buds and corolla), corolla, petals, etc. Is also included.

  The extract from cinnamon can be obtained by drying the raw material for extraction and then pulverizing the raw material as it is or using a crusher and subjecting it to extraction with an extraction solvent. Drying may be performed in the sun or using a commonly used dryer. Moreover, after performing pretreatment, such as degreasing, with a nonpolar solvent such as hexane, it may be used as an extraction raw material. By performing pretreatment such as degreasing, extraction treatment with a polar solvent of cinnabar can be performed efficiently.

  As the extraction solvent, it is preferable to use a polar solvent, and examples thereof include water and hydrophilic organic solvents. These are used alone or in combination of two or more at room temperature or a temperature below the boiling point of the solvent. It is preferable.

  Examples of water that can be used as the extraction solvent include pure water, tap water, well water, mineral spring water, mineral water, hot spring water, spring water, fresh water, and the like, and those subjected to various treatments. Examples of the treatment applied to water include purification, heating, sterilization, filtration, ion exchange, osmotic pressure adjustment, buffering, and the like. Therefore, the water that can be used as the extraction solvent in this embodiment includes purified water, hot water, ion exchange water, physiological saline, phosphate buffer, phosphate buffered saline, and the like.

  Examples of hydrophilic organic solvents that can be used as extraction solvents include lower aliphatic alcohols having 1 to 5 carbon atoms such as methanol, ethanol, propyl alcohol, and isopropyl alcohol; lower aliphatic ketones such as acetone and methyl ethyl ketone; 1,3-butylene. Examples thereof include polyhydric alcohols having 2 to 5 carbon atoms such as glycol, propylene glycol and glycerin.

  When using the liquid mixture of 2 or more types of polar solvents as an extraction solvent, the mixing ratio can be adjusted suitably. For example, when using a mixed solution of water and a lower aliphatic alcohol, it is preferable to mix 1 to 90 parts by volume of a lower aliphatic alcohol with respect to 10 parts by volume of water. When using a mixed solution, it is preferable to mix 1 to 40 parts by volume of a lower aliphatic ketone with 10 parts by volume of water, and when using a mixed solution of water and a polyhydric alcohol, water 10 It is preferable to mix 10 to 90 parts by volume of a polyhydric alcohol with respect to the volume part.

  The extraction treatment is not particularly limited as long as the soluble component contained in the extraction raw material can be eluted in the extraction solvent, and can be performed according to a conventional method. For example, the extraction raw material is immersed in an extraction solvent 5 to 15 times (mass ratio) of the extraction raw material, the soluble components are extracted at room temperature or under reflux, and then filtered to remove the extraction residue. A liquid can be obtained. When the solvent is distilled off from the obtained extract, a paste-like concentrate is obtained, and when this concentrate is further dried, a dried product is obtained.

  In addition, although the extract obtained as mentioned above can be used as an active ingredient of an anti-glycation agent as it is, a concentrated solution or a dried product is easier to use.

  In addition, since the extract from cinnamon moss has a unique odor, it can be purified for the purpose of decolorization, deodorization, etc. within a range that does not cause a decrease in its physiological activity. Since it is not used in a large amount when it is blended in, there is no practical problem even if it is not purified.

  The method for isolating and purifying acteoside from the extract obtained as described above, the concentrate of the extract or the dried product of the extract is not particularly limited, and can be performed by a conventional method. . For example, the extract is dissolved in a developing solvent, and subjected to column chromatography using a porous material such as silica gel or alumina, a porous resin such as a styrene-divinylbenzene copolymer or polymethacrylate, and the like. Examples include a method for collecting fractions. In this case, the developing solvent may be appropriately selected according to the stationary phase to be used. For example, when the extract is separated by normal phase chromatography using silica gel as the stationary phase, the developing solvent is chloroform / methanol / water = 10: 5: 1 etc. are mentioned. Further, the fraction containing acteoside obtained by column chromatography is subjected to any organic phase such as reverse phase silica gel chromatography using ODS, recrystallization, liquid-liquid countercurrent extraction, column chromatography using ion exchange resin, etc. You may refine | purify using a compound refinement | purification means.

  Since the extract and acteoside obtained from the above-described beetle have an excellent anti-glycation effect, they can be used as an active ingredient of an anti-glycation agent. The anti-glycation agent of this embodiment can be used for a wide range of uses such as pharmaceuticals, quasi drugs, and cosmetics. Here, acteoside is particularly preferred as an active ingredient of an anti-glycation agent because it exhibits a stronger anti-glycation effect than an extract from cinnamon.

  Here, the anti-glycation action which the extract from Actinoside and acteoside has is exhibited based on, for example, the formation inhibition action and / or the decomposition promotion action of final glycation products (AGEs). However, the anti-glycation action which the extract or acteoside from Buttercups has is not limited to the anti-glycation action exhibited based on the said action.

  Moreover, you may use the extract or acteoside from a Snapper as an active ingredient of an AGEs formation inhibitor or an AGEs decomposition promoter, respectively, using the AGEs formation inhibitory action or the decomposition promotion action which they have. Furthermore, the extract or acteoside from the umbilical oak is used for diabetic complications such as diabetic neuropathy, diabetic retinopathy, and diabetic nephropathy; Caused by arteriosclerosis; may be used as an active ingredient in prophylactic / therapeutic agents for diseases such as osteoporosis and osteoarthritis caused by glycation reaction of protein, and further decrease in skin elasticity caused by glycation reaction of protein It may be used as an active ingredient of preventive / improving agents such as dullness.

  In the present embodiment, any one of an extract from cinnamon or acteoside may be used as the active ingredient, or these may be mixed and used as the active ingredient. In the case of using an extract or acteoside from Buttercups as an active ingredient, the blending ratio may be appropriately adjusted depending on the degree of anti-glycation action of the extract or acteoside from Buttercups.

  The anti-glycation agent of the present embodiment may be composed only of an extract from cinnamon, acteoside or a mixture thereof, or may be an extract from cinnamon, acteoside or a mixture thereof.

  The anti-glycation agent of this embodiment is an arbitrary pharmaceutically acceptable carrier such as dextrin or cyclodextrin, and any other auxiliary agent such as powder, granule, tablet, liquid, etc. according to a conventional method. It can be formulated into a dosage form. In this case, as an auxiliary agent, for example, an excipient, a binder, a disintegrant, a lubricant, a stabilizer, a flavoring / flavoring agent, and the like can be used. The anti-glycation agent can be used by being blended with other compositions (for example, skin external preparations, cosmetic foods and drinks), and can also be used as an ointment, an external liquid, a patch and the like.

  When the anti-glycation agent of this embodiment is formulated, the content of the extract from cinnamon moss, acteoside, or a mixture thereof is not particularly limited, and can be appropriately set depending on the purpose.

  In addition, the anti-glycation agent of the present embodiment is used as an active ingredient, if necessary, by blending other natural extracts having an anti-glycation effect together with an extract from cinnamon, acteoside or a mixture thereof. Can do.

  Examples of the method for administering the anti-glycation agent of the present embodiment to a patient include transdermal administration, oral administration, and the like, and a suitable method for its prevention / treatment may be appropriately selected depending on the type of the disease. .

  In addition, the dose of the anti-glycation agent of the present embodiment may be appropriately increased or decreased depending on the type of disease, severity, individual differences among patients, administration method, administration period, and the like.

  The anti-glycation agent of this embodiment is a combination of diabetes mellitus such as diabetic neuropathy, diabetic retinopathy, diabetic nephropathy, etc. through the action of inhibiting the formation of AGEs and / or the action of promoting the degradation of AGEs possessed by an extract from cinnamon or acteoside The disease can be prevented or treated, and arteriosclerosis due to protein glycation can also be prevented or treated. In addition, the anti-glycation agent of the present embodiment reduces the skin elasticity and dullness caused by the glycation reaction of the protein through the action of inhibiting the formation of AGEs and / or the action of promoting the degradation of the AGEs possessed by the extract from Actinoside. Can be prevented or ameliorated. Furthermore, the anti-glycation agent of the present embodiment can also prevent or treat diseases such as osteoporosis and osteoarthritis caused by protein glycation reaction. However, the anti-glycation agent of the present embodiment can be used for all uses other than these uses, which are meaningful for exerting an AGEs formation inhibitory action and / or a decomposition promoting action.

  Moreover, since the anti-glycation action agent of this embodiment has the outstanding anti-glycation action, it is suitable for mix | blending with a skin external preparation or food-drinks, for example. In this case, the extract, acteoside, or a mixture thereof from cinnamon can be blended as it is, or an anti-glycation agent formulated from an extract, acteoside, or a mixture thereof from cinnamon can be blended.

  Here, the topical skin preparation is not limited in its category, and includes a wide range of skin cosmetics, quasi-drugs, pharmaceuticals, and the like used transdermally. Creams, milky lotions, beauty essences, lotions, packs, foundations, lip balms, bath salts, hair nicks, hair lotions, soaps, body shampoos and the like.

  There is no restriction on the category of food and drink, and it includes a wide range of foods such as general foods, health foods, and functional health foods that are taken orally.

  In addition, since the anti-glycation agent of the present embodiment has an excellent anti-glycation action, it is also suitably used as a reagent for research related to saccharification reaction, for example, research related to the formation mechanism and degradation mechanism of AGEs. be able to.

  In addition, although the anti-glycation agent of this embodiment is suitably applied to humans, as long as each effect is exhibited, animals other than humans (for example, mouse, rat, hamster, dog) , Cats, cattle, pigs, monkeys, etc.).

  Hereinafter, although a manufacture example and a test example are shown and this invention is demonstrated concretely, this invention is not restrict | limited to each following example at all.

[Production Example 1] Manufacture of an extract of flower portion of cinnamomum 1500 ml of 50% by weight ethanol (mass ratio of water to ethanol = 1: 1) is added to 150 g of a roughly pulverized flower portion of an antrum, and the mixture is gently stirred at 80 ° C. And filtered for 2 hours. Subsequently, 1500 mL of 50% by mass ethanol was added to the residue, kept at 80 ° C. for 2 hours with gentle stirring, and filtered while hot. The obtained filtrates were combined, concentrated under reduced pressure, and dried with a vacuum dryer to obtain an extract of flowering florets (71.4 g, sample 1).

[Production Example 2] Manufacture of Acteoside 8.9 g of the flower extract (sample 1) obtained from Production Example 1 was dissolved in a mixed solution of chloroform / methanol / water = 10: 5: 1 and silica gel (trade name) : Silica gel 60, manufactured by Merck & Co., Inc.) and was adsorbed on silica gel. Chloroform / methanol / water = 10: 5: 1 was passed as a moving bed through a glass column, the eluate was collected, and the solvent was distilled off to obtain a purified product (2.5 g, sample 2).

The purified product obtained by purification as described above was subjected to mass spectrum analysis, ¹ H-NMR analysis and ¹³ C-NMR analysis. The analysis results are shown below.

<Mass spectrum (ESI-MS)>
^{[M-H] - m /} z 623 ( theoretical _{value: C 29 H 36 O 15 -H} = 623)
[M + Na] ⁺ m / z 647 (theoretical value: C ₂₉ H ₃₆ O ₁₅ + Na = 647)

< ¹ H-NMR chemical shift δ (assigned hydrogen)>
6.69 (1H, d, J = 2.0 Hz, H-2), 6.67 (1H, d, J = 8.1 Hz, H-5), 6.55 (1H, dd, J = 2.0, 8.1Hz, H-6), 2.78 (2H, t-like, H-7), 4.01 (1H, overlapped, H-8a), 3.70 (1H, overlapped, H-8b), 7.05 (1H, d, J = 2.2 Hz, H-2 ' ), 6.77 (1H, d, J = 8.3 Hz, H-5 '), 6.94 (1H, dd, J = 2.2, 8.3Hz, H-6'), 7.58 (1H, d, J = 15.9 Hz, H -7 '), 6.27 (1H, d, J = 15.9 Hz, H-8'), 4.36 (1H, d, J = 7.8Hz, GlcH-1), 3.39 (1H, dd, J = 7.8, 8.1Hz , GlcH-2), 3.80 (1H, br.t, J = 9.0 Hz, Glc H-3), 4.88 (1H, br.t, J = 9.2 Hz, Glc H-4), 3.58 (1H, overlapped, Glc H-5), 3.48 (2H, overlapped, Glc H-6), 5.18 (1H, br.s, Rha H-1), 3.90 (1H, m, Rha H-2), 3.58 (2H, overlapped, Rha-H-3 and Rha H-5), 3.30 (1H, overlapped, Rha-H-4), 1.08. (3H, d, J = 6.1 Hz, Rha H-6)

< ¹³ C-NMR chemical shift δ (assigned carbon)>
131.1 (s, C-1), 116.1 (d, C-2), 146.5 (s, C-3), 144.3 (s, C-4), 116.8 (d, C-5), 121.0 (d, C -6), 36.4 (t, C-7), 72.0 (t, C-8), 127.3 (s, C-1 '), 114.9 (d, C-2'), 145.8 (s, C-3 ' ), 149.4 (s, C-4 '), 116.2 (d, C-5'), 122.9 (d, C-6 '), 147.7 (d, C-7'), 114.4 (d, C-8 ' ), 167.9 (s, C-9 '), 103.9 (d, Glc C-1), 75.9 (d, Glc C-2), 81.4 (d, Glc C-3), 70.7 (d, Glc C-4 ), 75.8 (d, Glc C-5), 62.1 (t, Glc C-6), 102.7 (d, Rha C-1), 72.1 (d, Rha C-2), 71.8 (d, Rha C-3 ), 73.5 (d, Rha C-4), 70.2 (d, RhaC-5), 18.3 (q, Rha C-6)

  From the above analysis results, it was confirmed that the compound obtained from the extract of the flowering part of the beetle was Acteoside represented by the following formula (I).

[Test Example 1] AGEs formation inhibitory action test For the cinnamon flower part extract (sample 1) and acteoside (sample 2) obtained in Production Examples 1 and 2, the final glycation product (AGEs) was analyzed as follows. The formation inhibitory action was tested.

  To a 96-well type I collagen-coated plate (Asahi Glass Co., Ltd.) 0.2 M D (-)-ribose prepared in PBS buffer and test samples (samples 1 and 2, see Table 1 for sample concentrations) 100 μL each of the mixed solution, PBS buffer only as a negative control, or 0.2 M D (−)-ribose solution prepared in PBS buffer as a positive control was added to each well and allowed to stand at 37 ° C. for 2 weeks. AGEs were formed. Two weeks later, the amount of AGEs was measured by an ELISA method using an anti-AGE antibody (manufactured by Transgenic) to evaluate the AGEs formation inhibitory action. From the obtained results, the AGE formation inhibition rate (%) was calculated by the following formula.

AGE formation inhibition rate (%) = {(BC) / (BA)} × 100
In the formula, A represents “absorbance at a wavelength of 405 nm of a negative control”, B represents “absorbance at a wavelength of 405 nm of a positive control”, and C represents “absorbance at a wavelength of 405 nm when a test sample is added”.
The results are shown in Table 1.

  As shown in Table 1, Samples 1 and 2 both inhibited the formation of AGEs in a concentration-dependent manner. Therefore, it was recognized that the extract of the flower part of the genus oleum and acteoside has an excellent AGEs formation inhibitory action.

[Test Example 2] AGEs degradation promoting action test With respect to the cinnamon flower part extract (sample 1) and acteoside (sample 2) obtained in Production Examples 1 and 2, the final glycation product (AGEs) was obtained as follows. The degradation promoting effect was tested.

  To a 96-well type I collagen-coated plate (Asahi Glass Co., Ltd.), 100 μL each of 0.2 M D (−)-ribose solution prepared in PBS buffer or PBS buffer alone as a negative control was added. AGEs were formed by allowing to stand at 2 ° C. for 2 weeks. Two weeks later, 100 μL each of a test sample solution prepared in PBS buffer (samples 1 and 2, see Table 2 for final concentrations), PBS buffer alone as a positive control, or PBS buffer alone as a negative control. After each addition, the mixture was further allowed to stand at 37 ° C for 2 weeks. After the reaction was completed, the amount of AGEs was measured by ELISA using an anti-AGE antibody (manufactured by Transgenic), and the AGEs degradation promoting action was evaluated. From the obtained results, the AGE decomposition rate (%) was calculated by the following formula.

AGE decomposition rate (%) = {(BC) / (BA)} × 100
In the formula, A represents “absorbance at a wavelength of 405 nm of a negative control”, B represents “absorbance at a wavelength of 405 nm of a positive control”, and C represents “absorbance at a wavelength of 405 nm when a test sample is added”.
The results are shown in Table 2.

  As shown in Table 2, both samples 1 and 2 promoted the degradation of AGEs. Therefore, it was recognized that the extract of the flower part of the hornbill and acteoside has an excellent AGEs degradation promoting action.

  The anti-glycation agent of the present invention is used for the prevention or treatment of diabetic complications such as diabetic neuropathy, diabetic retinopathy, diabetic nephropathy, prevention or treatment of arteriosclerosis caused by protein glycation reaction, protein glycation It can greatly contribute to the prevention or improvement of skin elasticity reduction and dullness caused by the reaction, and the prevention or treatment of diseases such as osteoporosis and osteoarthritis caused by the protein glycation reaction.

Claims (3)

  An anti-glycation agent characterized by containing an extract from rosaceae as an active ingredient.   An anti-glycation agent characterized by containing Acteoside as an active ingredient.   3. The anti-glycation agent according to claim 1 or 2, wherein the extract from the cynomolgus or the acteoside has an action of inhibiting the formation of final glycation products (AGEs) and / or an action of promoting degradation. .