JP2014108931A - Inhibitor of production of advanced glycation end product - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an inhibitor of production of advanced glycation end product comprising substances capable of inhibiting production of advanced glycation end product that is thought to be a causative substance of various lifestyle diseases, useful for prevention and/or treatment of lifestyle disease, particularly diabetes or diabetic complications, and having few adverse effects.SOLUTION: Provided is an inhibitor of production of advanced glycation end product comprising a Persicaria hydropiper (Polygonum hydropiper) f. purpurascens extract and/or a Persicaria hydropiper (Polygonum hydropiper) var. laetevirens extract as active ingredients.

Description

  TECHNICAL FIELD The present invention relates to a terminal glycation product production inhibitor containing a purple extract and / or an aotade extract as an active ingredient. The present invention also relates to a pharmaceutical composition for the prevention and / or treatment of diabetes or diabetic complications, which comprises a Murasakidae extract and / or an Aotade extract as an active ingredient.

  The phenomenon of browning when a mixture of amino acid and reducing sugar is heated is generally called the Maillard reaction, and is known in the food field as a phenomenon that occurs during heat treatment and storage of food. The Maillard reaction also occurs in vivo, and it was clear that glycosylation hemoglobin (HbA1c) was identified in vivo in 1968, leading to the progress of protein glycation with the progress of diabetes and aging. It was done. In recent years, advanced glycation end products (hereinafter also referred to as “AGEs”) in protein glycation reactions have been reported to be involved in the onset of lifestyle-related diseases such as diabetic complications and arteriosclerosis and the progression of aging. (Non-Patent Documents 1 to 3).

  The details of the protein saccharification reaction in vivo are not clear, but the amino group present in the protein reacts with the aldehyde group present in the reducing sugar to form a Schiff base, which then passes through a stable Amadori compound. It is believed that the transition to AGEs will occur after a long-term reaction.

  AGEs do not refer to specific substances, and the whole of them has not yet been elucidated, but the presence of various substances such as pentosidine, pyropyridine, and pyralin has been confirmed depending on the presence of fluorescence and cross-linking structure. The physicochemical characteristics of AGEs are that they are yellowish brown and fluorescent (mainly Ex: 370 nm, Em: 440 nm), and form crosslinks between proteins. However, AGEs that do not show fluorescence like pyralin and do not have a crosslinked structure have also been confirmed.

  The accumulation of these AGEs is not only due to skin aging, diabetic complications such as renal glomerular tissue sclerosis and renal arteriosclerosis, but also neurodegenerative diseases such as Alzheimer's disease, skin aging, bone aging, ocular aging, albumin It has been reported to be deeply involved in protein aging (Non-patent Document 4).

  Therefore, blocking the in vivo glycation reaction is expected to prevent the onset and progression of diabetic complications, age-related arteriosclerosis, myocardial abnormalities, and the like. For this reason, various saccharification reaction inhibitors and AGEs production inhibitors have been developed and studied.

  Patent Document 1 discloses a Maillard reaction inhibitor containing a compound such as aminoguanidine. Aminoguanidine is a substance exhibiting an anti-glycation effect that has been most studied in the development of pharmaceuticals in this technical field, but has been confirmed to have side effects such as liver damage and has not yet been put into practical use.

  In addition, Maillard reaction inhibitors using plant extracts such as Gambir and birch described in Patent Document 2 are known, but problems remain in terms of effectiveness and safety.

  Therefore, there is a demand for highly safe drugs that do not have serious side effects in suppressing the generation of AGEs involved in the onset and worsening of various diseases.

Japanese Examined Patent Publication No. 6-67827 JP 2005-035911 A

The Journal of Clinical Investigation, 1993, vol.91, pp.2470-2478 The New England Journal of Medicine, 1991, vol.325, pp.836-842 The Biochemical Journal, 2000, vol.350, pp.381-387 New Food Industry, 2011, vol.53, No.6

  The present invention includes substances that can suppress the generation of AGEs that are considered to be a causative substance of various lifestyle-related diseases, and is useful for the prevention and / or treatment of lifestyle-related diseases, particularly diabetes or diabetic complications. And it aims at providing the AGE production | generation inhibitor with few side effects.

  The inventors of the present invention have tried to screen from a plant that is a natural product and has a dietary experience with an excellent anti-glycation effect and has no side effects, and as a result of examining the plant extract and its components, the results of Murasakitade and Aotade The present invention was completed by finding a strong AGEs production inhibitory effect on the extract.

  That is, the present invention provides a terminal glycation product production inhibitor (hereinafter, also referred to as “AGEs production inhibitor of the present invention”) containing a purple extract and / or aotade extract as an active ingredient.

  The present invention also provides a pharmaceutical composition for the prevention and / or treatment of diabetes or diabetic complications (hereinafter referred to as “for the prevention / treatment of diabetes of the present invention”), which comprises a purple extract and / or an aotade extract as an active ingredient. Also referred to as “pharmaceutical composition”.

  The present invention further relates to a pharmaceutical composition for non-human animals comprising a Murasakidae extract and / or Aotade extract for the prevention and / or treatment of diabetes or diabetic complications in a non-human animal, the pharmaceutical composition or the present invention. A method for preventing and / or treating diabetes or diabetic complications in a non-human animal, comprising administering the AGEs production inhibitor of the present invention to a non-human animal, And the use of the AGE generation inhibitor of the present invention for the production of a supplement.

  AGEs are substances that have been suggested to be associated with various lifestyle-related diseases, and are particularly deeply involved in the onset and / or progression of diabetes and its complications. Therefore, the Murasakidae extract and the Aotade extract, which have been found to have the action of suppressing the generation of AGEs by the present inventors, are pharmaceutical compositions with few side effects for the prevention and / or treatment of diabetes or diabetic complications. Useful as an active ingredient.

FIG. 1 shows the rate of inhibition of saccharification reaction of aminoguanidine hydrochloride, purple lacquer extract and aotade extract. FIG. 2 shows the saccharification reaction inhibition rates of a plurality of fractions obtained by partition extraction from an ethanol extract of purple. FIG. 3 shows the saccharification reaction inhibition rates of a plurality of fractions obtained by partition extraction from the water extract of purple.

  The AGEs production inhibitor of the present invention contains a purple extract and / or an aotade extract as an active ingredient.

  Purple sardine (Persicaria hydropiper (Polygonum hydropiper) f. Purpurascens), also known as “Benitade” or “Akatade”, has a function to eliminate the smell and poison of raw food, so it can be eaten by using young shoots for sashimi Has been. On the other hand, Aotade (Persicaria hydropiper (Polygonum hydropiper) var. Laetevirens), also known as “Aitade”, has been eaten together with salted salmon, etc. A plant. In general, young shoots of Murasaki Kitade are circulated as food ingredients under the name of “red bean paste”, and the true leaves of Aotade are distributed under the name of “鮎 蓼”.

  The plant of purple and / or aotade used as a raw material for the extract may be in the flowering period or in the vegetative period, and may be in a raw state or a dried one. Good. Further, in order to increase the extraction efficiency, a plant body that has been appropriately shredded or pulverized, for example, a powdered one, may be used as an extraction raw material. As for the part of the plant used as a raw material, any of flowers, leaves, stems, roots, or whole plants containing these may be used as an extraction raw material. It is convenient to use such a part because the true leaves are easily available if there are any.

  The Murasakita extract and / or Aotade extract, which are the active ingredients of the AGEs production inhibitor and the pharmaceutical composition for the prevention / treatment of diabetes of the present invention, can be obtained using a general plant component extraction method. Examples of the extraction method include a hot water extraction method, a boiling extraction method, a soaking extraction method, a shaking extraction method, a Soxhlet extraction method, and a steam distillation method.

  Extraction solvents include water, lower alcohols such as methanol and ethanol, polyhydric alcohols such as propylene glycol and 1,3-butylene glycol, ketones such as acetone, esters such as diethyl ether, acetonitrile, and ethyl acetate. These can be selected from among these in consideration of the type of target plant, the subsequent processing, the intended use of the extract, and the like. If the organic solvent is not preferred because the extract is used for food applications, etc., only water should be used. Ethanol that can be easily removed after extraction is used alone or as a mixture with water in any proportion. You may do it. In one embodiment, the preferred extraction solvent is water or ethanol.

  The use amount of the extraction solvent is preferably from 1: 2 to 1:30 (plant raw material: extraction solvent), more preferably from 1: 3 to 1:20, and even more preferably from 1: 4 to 1:10 by weight ratio.

  The extraction temperature may be appropriately determined depending on each solvent, but in the case of water extraction, it is preferably 4 to 130 ° C, more preferably 25 to 110 ° C, and further preferably 40 to 100 ° C. When the extraction temperature is less than 4 ° C, the active ingredient tends to be difficult to extract, and when it exceeds 130 ° C, the active ingredient tends to be easily decomposed. In the case of ethanol extraction, 4 to 100 ° C is preferable, 10 to 80 ° C is more preferable, and 20 to 60 ° C is more preferable. When the extraction temperature is less than 4 ° C., the active ingredient tends to be difficult to extract, and when it exceeds 100 ° C., the active ingredient tends to be decomposed.

  In the case of water extraction, the extraction time is preferably 1 minute to 15 days, more preferably 3 minutes to 1 day, and further preferably 5 minutes to 1 hour. When the extraction time is less than 1 minute, the active ingredient tends to be difficult to extract, and when it exceeds 15 days, the active ingredient tends to be easily decomposed. In the case of ethanol extraction, 1 hour to 15 days is preferable, 12 hours to 10 days is more preferable, and 1 to 8 days is more preferable. When the extraction time is less than 1 hour, the active ingredient tends to be difficult to extract, and when it exceeds 15 days, the active ingredient tends to be easily decomposed.

  In the present specification, the term “extract” includes an extract obtained by various extraction methods using a solvent, a concentrated solution obtained by concentrating the extract, a diluted solution obtained by diluting the extract with an appropriate solvent, and the extraction. Solid extract (powdered or granular, etc.) obtained by removing the solvent from the liquid or its concentrated or diluted liquid by various methods such as heat drying, vacuum drying, freeze drying, etc., and solidifying agent or gel A solid or semi-solid extract obtained by addition of an agent is included. In one preferred embodiment, the Murasakita extract and / or Aotade extract used as an active ingredient in the AGEs production inhibitor and the pharmaceutical composition for preventing / treating diabetes of the present invention is in the form of a solid extract obtained by removing the solvent from the extract. It is.

  As an example of the method for extracting the Murasakidae extract and / or the Aotade extract, 2 to 30 parts by weight, preferably 3 to 10 parts by weight of water is added to 1 part by weight of the raw material plant, and the temperature is 40 to 90 ° C. Leave for 10 to 120 minutes, or boil at 100 ° C. for 5 to 15 minutes, or add 2 to 30 parts by weight, preferably 3 to 10 parts by weight of ethanol to 1 part by weight of the starting plant. The method of leaving still at 80 degreeC for 12 hours-10 days, Preferably leaving still at 20-60 degreeC for 1-8 days etc. are mentioned.

  The extract obtained from purple and / or aotade may be used as it is or may be subjected to a purification treatment. As the purification treatment, for example, a chromatographic method, an elution method using an ion exchange resin, partition extraction with a solvent, or the like can be employed alone or in combination. Examples of chromatographic methods include normal phase chromatography, reverse phase chromatography, thin phase chromatography, centrifugal liquid chromatography, high performance liquid chromatography, etc., and any of these or a combination thereof may be used. Can be mentioned. Solvents that can be used for partition extraction include ethyl acetate, hexane, 1-butanol and the like. In one embodiment, the purified extract that is extracted from the crude extract of purple and / or aotade into the ethyl acetate layer by partition extraction and recovered is the AGEs production inhibitor of the present invention or the pharmaceutical composition for prevention / treatment of diabetes of the present invention. It can exhibit excellent effects as an active ingredient.

  The AGEs production inhibitor of the present invention is only required to contain a Murasakidae extract and / or an Aotade extract as an active ingredient, and as long as the AGEs production inhibitory effect by the Murasakidae extract and / or Aotade extract is not hindered. It may contain an excipient or the like. Therefore, the ratio of the purple extract and / or aotade extract in the AGE production inhibitor of the present invention is not particularly limited. For example, in the case of using a solid extract, the AGEs production inhibitor of the present invention contains the Murasakitade extract and / or Aotade extract in an amount of 40% by weight, 45% by weight, 50% by weight, 60% by weight, 70% by weight. % Or more, 80% or more, 85% or more, 90% or more, 95% or more, or 98% or more. Or the AGEs production | generation inhibitor of this invention may consist only of a purple extract and / or an aotade extract.

  The AGEs production inhibitor of the present invention includes fluorescent AGEs such as pentosidine, crosslin, pyropyridine, and pyralin, carboxymethyllysine (CML), 3-deoxyglucosone-derived lysine dimer (DOLD), glyoxal-derived lysine dimer (GOLD), methyl Generation of various AGEs including non-fluorescent AGEs such as glyoxal-derived lysine dimer (MOLD) and imidazolone compounds can be suppressed. In one embodiment, the AGEs production inhibitor of the present invention is used for inhibiting the production of fluorescent AGEs.

The pharmaceutical composition for preventing / treating diabetes of the present invention contains, as with the AGEs production inhibitor of the present invention, a purple extract and / or aotade extract as an active ingredient.

The amount of Murasakidae extract and / or Aotade extract to be included as an active ingredient in the pharmaceutical composition for prevention / treatment of diabetes of the present invention can be appropriately set according to the intended dosage form, etc. When using a thing, it is usually about 1 to 98% by weight, preferably about 2 to 95% by weight, and more preferably about 3 to 90% by weight.

  As used herein, “treatment” of diabetes or diabetic complications includes preventing the progression of symptoms in a patient already suffering from diabetes or diabetic complications.

  The method for administering the pharmaceutical composition for preventing / treating diabetes of the present invention may be either oral administration or parenteral administration. The administration time is not particularly limited and may be any of before, during, after, and between meals. For example, a meal having a saccharide content of 5 g or more, or an amino acid and / or protein content of 20 g or more. It is preferably administered before meal, during meal, after meal and between meals, and before meal, during meal, after meal and between meals of meal containing sugar and amino acid and / or protein above the above content. More preferably it is administered. Examples of the amino acid include lysine, arginine, and the like. Among them, lysine and / or arginine, which is easily converted to AGEs, is preferably administered before, during, after, and between meals. Examples of the protein include ovalbumin, milk albumin, gliadin, milk casein, soybean casein, and the like. Among them, one or more selected from ovalbumin, milk albumin, and milk casein which are easily converted to AGEs are contained in the above content. It is preferably administered before, during, after and between meals. Here, “meal” generally means eating (drinking action) or eating or drinking food as a daily habit for taking the nutrients necessary for survival, but “meal” as used herein. The term "" means a set of food and drink taken in a single eating and drinking act.

  As used herein, “saccharides” shall mean monosaccharides and / or disaccharides. On the other hand, “sugar” refers to carbohydrates other than dietary fiber, and includes oligosaccharides, polysaccharides, sugar alcohols and the like in addition to sugars.

The dosage of the pharmaceutical composition for preventing / treating diabetes according to the present invention is appropriately selected according to conditions such as the degree of diabetes or its complications, the degree of other diseases, age, and sex, but suppresses the generation of AGEs. An amount that can be administered (hereinafter also referred to as “AGEs suppression effective amount”) may be administered. An effective amount for inhibiting the generation of AGEs can be appropriately determined using methods well known to those skilled in the art (including various non-clinical and / or clinical trials).

  The pharmaceutical composition for preventing / treating diabetes according to the present invention may be one in which an AGEs production-inhibiting effective amount of Murasakitade extract and / or Aotade extract is administered at once, and is administered in multiple portions at intervals. It may be what you do. In the case of administration in multiple doses, the total amount of purple extract and / or aotade extract administered per day may be an effective amount for inhibiting the generation of AGEs, and is preferably administered according to the number of meals. .

  The pharmaceutical composition for preventing / treating diabetes according to the present invention can be administered to either a person suffering from diabetes or diabetic complications or a healthy person, but suffering from diabetes having a high association with AGEs. It is preferably administered to a person. A person suffering from diabetes refers to a person whose hemoglobin A1c is 6.1% (JDS value) or more and falls under any of the following (1) to (3): (1) Fasting blood glucose level 126 mg / dL or more, (2) Adequate blood glucose level (whether fasting or after eating) is 200 mg / dL or more, and (3) 2 hours after glucose loading is 200 mg / dL or more. Details of the diagnostic criteria for diabetes are described in the “Report of the Committee on Diabetes Classification and Diagnostic Criteria” by the Japanese Diabetes Association (available from the association's website).

  Further, the pharmaceutical composition for preventing / treating diabetes of the present invention comprises diabetic retinopathy, diabetic nephropathy, diabetic neuropathy, diabetic vascular complications, arteriosclerosis, renal failure, Alzheimer's disease, neurodegeneration, which develops with diabetes. It can be used for the prevention and / or treatment of diabetic complications such as diseases and cancer. Among these diabetic complications, it is preferably used for the prevention and / or treatment of diabetic retinopathy, diabetic nephropathy and / or diabetic neuropathy with a high incidence.

  Furthermore, diabetes and its complications can be effectively prevented by administering the pharmaceutical composition for preventing / treating diabetes of the present invention to healthy individuals.

  The pharmaceutical composition for the prevention / treatment of diabetes of the present invention is not limited to the Murasaki Kitade extract and / or the Aotade extract so long as the effect of preventing and / or treating diabetes or diabetic complications is prevented. In addition to the extract, ingredients such as excipients, stabilizers, preservatives, buffering agents, flavoring agents, suspending agents, emulsifiers, flavoring agents, solubilizing agents, coloring agents, and thickeners are added. Various dosage forms can be provided.

  Examples of the dosage form of the pharmaceutical composition for prevention / treatment of diabetes of the present invention include tablets (orally disintegrating tablets, chewable tablets, effervescent tablets, dispersible tablets, dissolving tablets), capsules, granules (expanded granules), powders, Oral solution (elixir, suspension, emulsion, limonade), syrup (syrup), oral jelly, oral tablet (troche, sublingual, buccal, adhesive tablet, gum), oral Spray, oral semi-solid preparation, mouthwash, injection (infusion solution, implantable injection, continuous injection), dialysis agent (peritoneal dialysis agent, hemodialysis agent), inhalant (inhalation powder) , Inhalants, aerosols), suppositories, rectal semisolids, enemas, eye drops, eye ointments, ear drops, nasal drops (nasal powders, nasal drops), vaginal tablets, Vaginal suppository, external solid preparation (external powder), external liquid (liniment, lotion), spray (External aerosols, pump sprays), ointments, creams, gels, patches (tape, cataplasms), and the like. Among these, tablets, capsules, granules, powders, oral solutions, syrups, oral jelly, oral tablets, oral sprays, oral semisolids and mouthwashes are easy to administer. From the viewpoint of being easily absorbed in the living body, tablets, capsules, granules, and powders are more preferable.

  Further, the pharmaceutical composition for preventing / treating diabetes of the present invention can be applied to herbal medicine-related preparations such as extracts, pills, spirits, soaking agents, decoction, teas, tinctures, fragrances, and flow extracts. It can also be used in a dosage form to which an AGEs production inhibitor is added. These dosage forms are appropriately selected according to conditions such as the degree of diabetes or its complications, the degree of other diseases, age, and sex.

  Furthermore, the pharmaceutical composition for preventing / treating diabetes according to the present invention may further contain other drugs having an AGEs production inhibitory effect. Examples of these drugs include insulin resistance improving drugs, dyslipidemic drugs, angiotensin II type 1 receptor antagonists, angiotensin converting enzyme inhibitors, metformin, and the like.

  In addition, the pharmaceutical composition for preventing / treating diabetes of the present invention may further contain a natural product or a naturally-derived material having an AGEs production inhibitory effect. Examples of these natural products or naturally-derived substances include herbs such as dokudami, hawthorn, chamomile, and grape leaves, cherry blossoms, corn stigma and stigmas, plantain seeds, maronier, peonies, rose flowers, plum fruit , Mountain grape, purple chrysanthemum flower, wheat germ, wheat germ-derived polyamine, mangosteen and the like.

  The AGEs production inhibitor of the present invention or its active ingredient, Murasakita extract and / or Aotade extract, can also be used to prevent and / or treat diabetes or diabetic complications in non-human animals. Therefore, in one aspect of the present invention, a pharmaceutical composition for non-human animals (hereinafter referred to as the present invention) comprising a Murasakida and / or Aotade extract for the prevention and / or treatment of diabetes or diabetic complications in a non-human animal. Inventive non-human animal pharmaceutical composition). In addition, a method for preventing and / or treating diabetes or diabetic complications in a non-human animal, comprising administering the non-human animal pharmaceutical composition or the AGEs production inhibitor of the present invention to a non-human animal is also provided. Examples of non-human animals include livestock such as cows, horses, pigs, sheep, goats and chickens, and animals raised as pets such as dogs and cats.

  The amount of Murasakida extract and / or Aotade extract to be contained in the pharmaceutical composition for non-human animals of the present invention, the method of administering the pharmaceutical composition, the timing of administration, the dosage and the dosage form, and the Murasakiitade extract in the pharmaceutical composition And / or other drugs, natural products and naturally derived substances that can be used in combination with the Aotade extract, and the administration method, administration time, dosage, etc. when administering the AGEs production inhibitor of the present invention to a non-human animal, The same as described above for the “pharmaceutical composition for preventing / treating diabetes of the present invention”.

  Furthermore, the AGEs production | generation inhibitor of this invention can also be used as a food additive. Therefore, the functional food / beverage products which suppress the production | generation of AGEs can be provided by mix | blending or adding the AGEs production | generation inhibitor of this invention to food / beverage products. Moreover, the supplement which suppresses AGEs production | generation can also be manufactured by using the AGEs production | generation inhibitor of this invention as a component. In this case, excipients and the like can be appropriately blended according to the dosage form of the supplement to be produced.

  EXAMPLES Hereinafter, although an Example is given and this invention is demonstrated concretely, this invention is not limited to these description.

Example 1
Extraction of purple (degradation with ethanol)
Purple shoots (cotyledons) (400 g) were soaked in ethanol (2.0 L), left to stand at 25 ° C. for 7 days for extraction, and separated into an extract and a residue by filtration to obtain an extract (material) And the weight ratio of the extraction solvent is 1: 5). The extract was dried under reduced pressure to obtain an extract (11.03 g). Hereinafter, such an ethanol extract is referred to as a purple extract (E).

Extraction of Murasakikatade (extraction with water)
Water (1.2L) is added to purple shoots (cotyledons) (300 g), boiled at 100 ° C. for 5 minutes, extracted, and separated into an extract and a residue by filtration to obtain an extract (raw material and extract) The weight ratio of the solvent is 1: 4). The extract was dried by lyophilization to obtain an extract (11.30 g). Hereinafter, such a hot water extract is referred to as a purple extract (W).

Aotade extraction (extraction with ethanol)
The real leaves and petioles (334 g) of Aotade shredded to a size of 2 to 10 mm square are immersed in ethanol (1.67 L), left to stand at 25 ° C. for 7 days, extracted, and filtered with the extract. Divided into residues, an extract was obtained (weight ratio of material to extraction solvent is 1: 5). The extract was dried under reduced pressure to obtain an extract (11.75 g). Hereinafter, such an ethanol extract is referred to as an Aotade extract (E).

Aotade extraction (water extraction)
Water (0.6 L) is added to real leaves and petioles (150 g) shredded to a size of 2 to 10 mm square, boiled at 100 ° C. for 5 minutes, extracted, and the extract and residue are filtered. An extract was obtained (weight ratio of material to extraction solvent was 1: 4). The extract was dried by lyophilization to obtain an extract (9.25 g). Hereinafter, such a hot water extract is referred to as an Aotade extract (W).

Measurement of Saccharification Reaction Inhibition Rate Using Murasakidea extract and Aotade extract, the effect on AGE formation was tested by the following method. To 0.9 mL of 67 mM phosphate buffer (pH 7.2) containing 10 wt% D-glucose (manufactured by SIGMA-ALDRICH) and 1 wt% bovine serum albumin (fraction V, manufactured by SIGMA-ALDRICH), 0.1 mL of a sample solution having the composition shown was added, and the mixture was allowed to stand at 60 ° C. for 2 days. The fluorescence intensity (excitation wavelength: 370 nm, fluorescence wavelength: 440 nm) of the reaction solution was measured using Varioskan Flash (manufactured by Thermo), and this value was defined as a1 in the saccharification reaction inhibition rate calculation formula shown below. B1 to d1 in the calculation formula were also reacted under the same conditions, the fluorescence intensity was measured, and the saccharification reaction inhibition rate was calculated.

Saccharification reaction inhibition rate (%) = 100 − {(a1−b1) / (c1−d1)} × 100
a1: Sample solution of each test section 0.1 mL + (10 wt% D-glucose + 1 wt% bovine serum albumin + 67 mM phosphate buffer solution) 0.9 mL
b1: 0.1 mL of sample solution in each test group + 0.9 mL of 67 mM phosphate buffer
c1: Solvent 0.1 mL in each test group + (10 wt% D-glucose + 1 wt% bovine serum albumin + 67 mM phosphate buffer) 0.9 mL
d1: Solvent 0.1 mL + 67 mM phosphate buffer 0.9 mL in each test group

ＤＭＳＯ：ジメチルスルホキシド

DMSO: Dimethyl sulfoxide

  The Murasakidae extract and / or Aotade extract, which are the active ingredients of the AGEs production inhibitor of the present invention, showed a glycation reaction inhibition rate equal to or higher than that of aminoguanidine hydrochloride. The results are shown in Table 2 and FIG.

Example 2
Dispersion extraction of purple purple extract (E) The purple purple extract (E) produced by the same method as in Example 1 was extracted in the order of a low polarity solvent with three kinds of solvents having different polarities.
Disperse 1.71 g of purple purple extract (E) (dry solid) in 100 mL of water, stir for 10 minutes using 100 mL of hexane, perform distribution, and further extract the water-soluble part with hexane for a total of 3 times The hexane transfer part and the water-soluble part were separated. The obtained hexane transition part was evaporated under reduced pressure to obtain 0.54 g of a hexane-soluble part (E). Next, the obtained water-soluble part was extracted three times using the same amount of ethyl acetate as the water-soluble part (stirring 10 minutes / times), and divided into an ethyl acetate transfer part and a water-soluble part. . The obtained ethyl acetate transition part was evaporated under reduced pressure to obtain 50.98 mg of ethyl acetate soluble part (E). Further, the obtained water-soluble part was extracted three times using the same amount of 1-butanol as that of the water-soluble part (stirring 10 minutes / times), and divided into a 1-butanol transition part and a water-soluble part. The obtained 1-butanol transition part was evaporated under reduced pressure to obtain 0.25 g of 1-butanol soluble part (E). The water-soluble part finally obtained by this operation of partition extraction was lyophilized to obtain 0.74 g of a water-soluble part (E).

Measurement of inhibition rate of saccharification reaction Purple extract (E) and hexane soluble part (E), ethyl acetate soluble part (E), 1-butanol soluble part (E), water soluble obtained by partition extraction Using the part (E), the effect on the AGE formation was tested by the following method.
0.5 M phosphorus containing 0.1 MDL-glyceraldehyde (manufactured by Hanai Co., Ltd.), 5 mg / mL each of Boc-Lys-OH (manufactured by Watanabe Chemical Co., Ltd.), Boc-Arg-OH (manufactured by Watanabe Chemical Co., Ltd.) 0.25 mL of a sample solution having the composition shown in Table 3 was added to 1.25 mL of an acid buffer (pH 7.2), and the mixture was allowed to stand at 37 ° C. for 7 days. The fluorescence intensity (excitation wavelength: 380 nm, measurement range: 390-900 nm) of the reaction solution was measured using Varioskan Flash (manufactured by Thermo), and this value was defined as a2 in the saccharification reaction inhibition rate calculation formula shown below. Fluorescence intensity was measured by reacting b2 to d2 in the calculation formula under the same conditions. For e2 and f2, the fluorescence intensity was measured immediately after mixing, and the saccharification reaction inhibition rate was calculated.

Saccharification reaction inhibition rate (%) = 100 − {(a2−b2) − (e2 + f2) / (c2−d2) − (e2 + f2)} × 100

a2: 0.25 mL of sample solution in each test group + (1.25 mL of 0.1 MDL-glyceraldehyde + 5 mg / mL Boc-Lys-OH + 5 mg / mL Boc-Arg-OH + 0.5 M phosphate buffer)
b2: 0.25 mL of sample solution in each test group + 1.25 mL of 0.5 M phosphate buffer
c2: 0.25 mL of the solvent in each test group + (0.1 MDL-glyceraldehyde + 5 mg / mL Boc-Lys-OH + 5 mg / mL Boc-Arg-OH + 0.5 M phosphate buffer) 1.25 mL
d2: 0.25 mL of solvent in each test group + 1.25 mL of 0.5 M phosphate buffer
e2: 5 mg / mL Boc-Lys-OH + 5 mg / mL Boc-Arg-OH + 0.5 M phosphate buffer f2: 0.1 MDL-glyceraldehyde + 0.5 M phosphate buffer

ＤＭＳＯ：ジメチルスルホキシド

DMSO: Dimethyl sulfoxide

  The ethyl acetate-soluble part (E) obtained by performing partition extraction on the Murasakidae extract (E), which is an active ingredient of the AGEs production inhibitor of the present invention, is superior in saccharification compared to other fractions. The reaction inhibition rate was shown. The results are shown in Table 4 and FIG.

Example 3
Dispersion extraction of purple purple extract (W) The purple purple extract (W) produced by the same method as in Example 1 was extracted in the order of the solvents having the least polarity with three kinds of solvents having different polarities.
Murasaki Akitade extract (W) (dry solid) 3.18g was dissolved in 100mL of water, and after stirring for 10 minutes using 100mL of hexane, the water-soluble part was further extracted three times with hexane. The hexane transfer part and the water-soluble part were separated. The obtained hexane transition part was evaporated under reduced pressure to obtain 7.30 mg of a hexane-soluble part (W). Next, the obtained water-soluble part was extracted three times using the same amount of ethyl acetate as the water-soluble part (stirring 10 minutes / times), and divided into an ethyl acetate transfer part and a water-soluble part. . The resulting ethyl acetate transition part was evaporated under reduced pressure to obtain 1.04 g of ethyl acetate soluble part (W). Further, the obtained water-soluble part was extracted three times using the same amount of 1-butanol as that of the water-soluble part (stirring 10 minutes / times), and divided into a 1-butanol transition part and a water-soluble part. The obtained 1-butanol transition part was evaporated under reduced pressure to obtain 0.68 g of 1-butanol soluble part (W). The water-soluble part finally obtained by this operation of partition extraction was lyophilized to obtain 1.38 g of a water-soluble part (W).

Measurement of inhibition rate of saccharification reaction Murasakikita extract (W) and hexane soluble part (W), ethyl acetate soluble part (W), 1-butanol soluble part (W), water soluble obtained by partition extraction Using part (W), the effect on AGE formation was tested using the sample solution having the composition shown in Table 5 in the same manner as the measurement of the saccharification reaction inhibition rate of the purple extract (E). .

  The ethyl acetate soluble part (W) obtained by partition extraction of the Murasakidae extract (W), which is an active ingredient of the AGEs production inhibitor of the present invention, is superior in saccharification compared to other fractions. The reaction inhibition rate was shown. The results are shown in Table 6 and FIG.

Production of powder The raw materials shown in Table 7 were mixed to produce a powder. The powder is one form of the AGE production inhibitor of the present invention or the pharmaceutical composition for preventing / treating diabetes of the present invention.

Preparation of tablets After mixing the raw materials shown in Table 8, using a continuous tableting machine (Piccola B-10 / RIVA), a die (φ8mm, R12mm), weight 150-200mg per tablet, rotation Tablets were produced directly by tableting under the tableting conditions of 12 rpm and tableting pressure of 4 kN. The tablet is one form of the AGE production inhibitor of the present invention or the pharmaceutical composition for preventing / treating diabetes of the present invention.

Production of oral solution The raw materials shown in Table 9 were dissolved in distilled water to make a total volume of 500 mL to produce an oral solution. The oral solution is one form of the pharmaceutical composition for preventing / treating diabetes of the present invention.

Claims (10)

  A terminal saccharification product production inhibitor containing a purple peach extract and / or an aotade extract as an active ingredient.   A pharmaceutical composition for the prevention and / or treatment of diabetes or diabetic complications, comprising an extract of purple and / or aotade as an active ingredient.   The pharmaceutical composition according to claim 2, comprising an effective amount of glycation end product production-inhibiting purple extract and / or aotade extract.   The pharmaceutical composition according to claim 2 or 3, wherein the diabetic complication is one or more diseases selected from the group consisting of diabetic retinopathy, diabetic nephropathy, diabetic neuropathy, diabetic vascular complication and arteriosclerosis. .   The saccharide content is 5 g or more, and the amino acid content and / or protein content is 20 g or more, which is administered before, during, after or between meals. A pharmaceutical composition according to any one of the above.   An agent selected from the group consisting of tablets, capsules, granules, powders, oral solutions, syrups, oral jelly, oral tablets, oral sprays, oral semisolids, mouthwashes, injections and inhalants A pharmaceutical composition according to any of claims 2 to 5, which is in the form.   A pharmaceutical composition for a non-human animal, comprising an extract of purple and / or aotade as an active ingredient for the prevention and / or treatment of diabetes or diabetic complications in a non-human animal.   A method for preventing and / or treating diabetes or diabetic complications in a non-human animal, comprising administering the glycation end product production inhibitor according to claim 1 or the pharmaceutical composition according to claim 7 to the non-human animal. .   The manufacturing method of functional food / beverage products characterized by mix | blending or adding the glycation end product production inhibitor of Claim 1 to food / beverage products.   Use of the terminal glycation product production inhibitor according to claim 1 for the production of a supplement.

Images