JP4954564B2 - Formulation for degrading protein-reducing sugar cross-linking substance which is Maillard reaction product - Google Patents

Description

  The present invention relates to a preparation that degrades a protein-reducing sugar cross-linking substance that is a Maillard reaction product.

Maillard reaction is known as a non-enzymatic reaction between glucose or other reducing sugar and an amino acid having an amine group or a protein having an amino acid residue thereof. In the Maillard reaction, glucose or other reducing sugars are attached to the free amino group of the protein (initial glycosylation) to form a Schiff base at an early stage, which gradually undergoes Amadori rearrangement, resulting in an Amadori product (initial glycosyl). Product). The Amadori product further proceeds, and a depolymerized and rearranged reaction is repeated to produce a cross-linked compound (late glucosylation product) (late stage). This cross-linked polymerized compound is called Advanced Glycation End Products (AGEs), and its physical properties are reduced in solubility, less susceptible to the action of protease, fluorescent color is developed and turns brown. This Maillard reaction was first discovered from the appearance of brown pigment in foods, but later studies revealed that it was also carried out in vivo. For example, it is reported in Japanese Patent Publication No. 06-67827 that reaction products with blood hemoglobin and albumin are composed, and that similar reactions occur with lens proteins, collagen, and neuroproteins. (Patent Document 1).
Japanese Patent Publication No. 6-67827

As shown in FIG. 1, the AGEs are formed by binding glucose or other reducing sugar to the side chain amine group of the protein, leaving the water molecule and changing the glucose moiety to diketone, and then binding other protein. (Non-Patent Document 1). By accumulating this AGEs, diabetic complications such as nephropathy, neuropathy, retinopathy, cataract, microvascular disorder, arteriosclerosis, glomerulonephritis, circulatory failure, senile cataract, osteoarthropathy, Diseases associated with aging occur, such as periarticular stiffness, joint sclerosis, scleroderma, senile plaques, and senile dementia. Until now, in order to prevent or ameliorate the above-mentioned various diseases, methods for orally or parenterally administering a component having an action of inhibiting the generation of AGEs to the human body have been studied. Examples include butyric acid derivatives (Patent Document 2), citrus volatile oils (Patent Document 3), aminoguanidine, and α-hydrazinohistidine (Patent Document 4).
Sara V., Xin Z., et.al., Nature, vol. 382, 18, 275-278 (1996) Special Table 2002-541139 JP 2004-35424 A Japanese Patent Publication No. 06-67827

However, the above components only inhibit the generation of AGEs, do not have the property of metabolizing or disappearing the already generated AGEs, it is possible to prevent diabetic complications and age-related diseases But it is impossible to treat them. Therefore, in order to treat various diseases, it is necessary to find a component having an action of degrading generated and accumulated AGEs. As described above, AGEs are substances in which proteins are cross-linked via a reducing sugar, and the action of degrading AGEs is the action of cleaving the cross-linked portion of the protein-reducing sugar cross-linking substance, such as Examples of the component having an action include N-phenanthyl thiazolium bromide (Non-patent Document 1) and extracts of plants such as American witch hazel, apricot and yew (Patent Document 5). However, no active ingredient having an action that exceeds the effectiveness of N-phenanthyl thiazolium bromide has been found so far, and the effectiveness of N-phenanthyl thiazolium bromide exceeds the effectiveness of each plant extract. There was nothing.
Japanese Patent Laid-Open No. 2002-241299

  Accordingly, an object of the present invention is to newly provide an active ingredient having an action of cleaving a cross-linked portion of a protein-reducing sugar cross-linking substance superior to that of N-phenanthyl thiazolium bromide.

  In the present inventors, rosmarinic acid or a salt thereof, myricetin, epigallocatechin, epigallocatechin gallate, baicalin and baicalein have an action of cleaving the cross-linked portion of the protein-reducing sugar cross-linking substance, and the action is Found the same or better than N-phenanthyl thiazolium bromide, and provided a preparation that decomposes the protein-reducing sugar cross-linking substance, which is a new Maillard reaction product, to solve the above problems. .

  The preparation of the present invention is capable of degrading the protein-reducing sugar cross-linking substance, which is a Maillard reaction product, and this action has the same or higher efficacy as a component having the same action known so far. Indicated. By applying this preparation to the human body, it is possible to treat diabetic complications and diseases associated with aging. In particular, the preparation in the form for oral administration is very effective for the treatment of diabetic complications, and the form for transdermal administration is very effective for the treatment of aging-related diseases.

  Rosmarinic acid used in the present invention is a compound represented by the following formula 1. The salt is selected from sodium, potassium, calcium and magnesium salts.

：式１

: Formula 1

  Myricetin used in the present invention is a compound represented by the following formula 2.

：式２

: Formula 2

  Epigallocatechin used in the present invention is a compound represented by the following formula 3.

：式３

: Formula 3

  Epigallocatechin gallate used in the present invention is a compound represented by the following formula 4.

：式４

: Formula 4

  Baicalin used in the present invention is a compound represented by the following formula 5.

：式５

: Formula 5

  The baicalein used in the present invention is a compound represented by the following formula 6.

：式６

: Formula 6

  Each of the above-mentioned compounds can be obtained by individually extracting and purifying from Lamiaceae plants such as Mannenrou, Aojiso, Chilemiso, Kawamidori, Lifelong Grass, etc., bayberry, Koganabana, Cha (Ryokucha, Kocha, Oolongcha, etc.). Examples of the extraction method include a solvent extraction method mainly using water or an organic solvent, or a supercritical fluid extraction method. As a purification method, a chromatogram using a synthetic adsorbent or a resin is mainly used. In addition, a separation-purification method by liquid-liquid distribution, solid-liquid distribution, or the like is used. Each plant extract obtained by solvent extraction is adsorbed onto a chromatographic column packed with a synthetic adsorbent, and the target compound is drained from the target fraction using a specific elution organic solvent to remove the solvent. To get more. Each of the above-mentioned compounds can be obtained using a known organic chemical synthesis method, but it is preferably obtained by extraction from a plant in consideration of the yield and the complexity of the production process. Further, in the process of extracting and purifying each of the above compounds from a plant, a plant extract containing the above compound obtained as a main component by adding a process of further enrichment or a process of increasing the concentration of only a specific component and omitting the process of purification. This form can also be used in the present invention in the same manner as the compound.

  The content of each compound in the preparation is in the range of 0.001 to 50% by mass, and is preferably in the range of 0.01 to 10% by mass in view of the expected resolution of the crosslinked product. If the content is less than 0.001% by mass, the resolution is insufficient, and if the content exceeds 50% by mass, excessive resolution will be imparted, and only the manufacturing cost will increase, and the formulation will be composed Depending on the type of composition component to be used, the stability of the preparation itself may be lowered.

  The dosage form of the preparation of the present invention is not particularly limited as long as it can be administered orally or parenterally. Capsule, powder, granule, solid, liquid, gel, foam, emulsion, cream Forms, ointments, sheets, mousses, powder dispersions, multilayers, aerosols, and the like. One particularly preferred form is the form of oral administration, and specific examples of the dosage form include solutions (including forms used by spraying), granules, powders, tablets, pellets, and oral films. And capsules. Another particularly preferred form is a form of transdermal administration, and specific examples of the dosage form include lotions represented by cosmetics and quasi drugs (including whitening and anti-aging), Latex (including whitening and anti-aging), cream (including whitening and anti-aging), lotion (including whitening and anti-aging), pack (including whitening and anti-aging), soap, In addition to body shampoos, facial cleansers, lipsticks, foundations, etc., ointments and patches, lotions, poultices, liniments and the like represented by pharmaceuticals and quasi-drugs.

  In addition to the above-mentioned compounds that are essential active ingredients, the preparation of the present invention further includes Maillard reaction inhibitory ingredients, whitening ingredients, tanning ingredients, moisturizing ingredients, cell activation ingredients, antioxidant ingredients, UV protection ingredients, and UV absorption promotion. Extracts derived from ingredients, astringent ingredients, anti-inflammatory ingredients, antiseptic ingredients, MMP activity inhibitory ingredients, peeling ingredients, hair growth / hair nourishing ingredients, plant raw materials / animal raw materials / microbe raw materials / other natural raw materials, pH A base that is acceptable for producing regulators, chelating agents, thickeners, foaming agents, propellants, surfactants, fragrances, deodorizing / deodorizing ingredients, other pharmaceuticals / quasi-drugs, cosmetics, etc. Can be blended. In particular, by incorporating a Maillard reaction inhibiting component at the same time, it is possible to control and prevent all the diseases that occur due to the Maillard reaction. Although content in a formulation is not specifically limited, Usually, the density | concentration range of 0.0001 to 50 weight% is common.

  Hereinafter, the present invention will be described in more detail with reference to examples, but the present invention is not limited thereto.

Protein-reducing sugar cross-linking substance degradation test of each compound Rosmarinic acid (Funakoshi Co., Ltd.), Myricetin (Funakoshi Co., Ltd.), Epigallocatechin (Kurita Kogyo Co., Ltd.), Epigallocatechin Galate (Kurita Kogyo Co., Ltd.) A DMSO solution containing 1 mM of baicalin (manufactured by Toyobo Co., Ltd.) or baicalein (manufactured by Wako Co., Ltd.) was prepared and prepared for use in the test. The test method was the method described in Sara V., Xin Z., et.al., Nature, vol. 382, 18, 275-278 (1996). 100 mL DMSO solution containing 100 mM 1-phenyl-1,2-propanedione, a model of protein-reducing sugar cross-linking substance, 100 μL of each of the above compounds, and 1.5 mL of 500 mM disodium phosphate aqueous solution (pH 7.4) 800 μL The mixture was mixed in a plastic tube and incubated at 37 ° C. for 3 hours with a shaker to react. Next, the amount of benzoic acid contained in the reaction solution was analyzed and quantified using HPLC. The resolution of each compound was calculated based on the quantitative result and 1 molecule of 1-phenyl-1,2-propanedione being decomposed to produce 1 molecule of benzoic acid. N-phenanthyl thiazolium bromide was used as a positive control, HPLC analysis was used Column: RP-18 (manufactured by Kanto Chemical Co., Inc.), mobile phase: 40% methanol-0.1% TFA, flow rate: 1.0 mL / min, column temperature : 40 ° C., detection: UV 210 nm. Table 1 shows the resolution of each compound when the resolution when N-phenanthyl thiazolium bromide was added was taken as 100%.

Results of protein-reducing sugar crosslinking substance degradation test for each compound
[Table 1]
200% rosmarinic acid
Myricetin 200%
Epigallocatechin 168%
Epigallocatechin gallate 175%
Baicalin 108%
Baikarain 98%
* When the resolution of N-phenanthyl thiazolium bromide is 100%.
As shown in Table 1, rosmarinic acid, myricetin, epigallocatechin, epigallocatechin gallate, baicalin, and baicalein used in the present invention were compared to the resolution of N-phenanthylthiazolium bromide, which is a positive control, as well as baicalin and Compounds other than baicalein showed significantly higher 1-phenyl-1,2-propanedione resolution. Baicalin and baicalein showed the same resolution as N-phenanthyl thiazolium bromide. Therefore, rosmarinic acid, myricetin, epigallocatechin, epigallocatechin gallate, baicalin and baicalein may have the same or superior degradation action of protein-reducing sugar cross-linking substance compared to N-phenanthyl thiazolium bromide. found.

(Manufacturing examples of various preparations)
Various dosage forms according to the present invention were prepared. Although the formulation example is shown below, this invention is not necessarily limited to these.

(Prescription Example 1) Capsule
Rosmarinic acid 8.0g
Magnesium ascorbate 0.5g
DL-α-tocopherol acetate 0.5g
Silicone oil 30.0g
Tween 80 0.05g
Corn starch 48.95g
The above ingredients were mixed uniformly and 200 mg was filled into a gelatin capsule to produce a capsule.

(Prescription Example 2) Tablet
Myricetin 100.0mg
Cornstarch 50.0mg
Hydroxypropyl methylcellulose 150.0mg
Carboxymethylchitin 30.0mg
The above ingredients were uniformly mixed and tableted to produce a tablet.

(Prescription Example 3) Granule
Epigallocatechin gallate 50.0mg
Carboxymethylcellulose 180.0mg
Propylene glycol 20.0mg
Sodium alginate 30.0mg
Sucrose 10.0mg
The above ingredients were uniformly mixed and granulated to produce granules.

(Formulation Example 4) Milk Mass%
Epigallocatechin 3.5
Squalane 5.0
Olive oil 5.0
Jojoba oil 5.0
Cetyl alcohol 1.5
Glycerol monostearate 2.0
Polyoxyethylene (20) cetyl ether 3.0
Polyoxyethylene (20) soorbitan monooleate 2.0
1,3-butylene glycol 1.0
Glycerin 2.0
P-Hydroxybenzoate ester
Residue with 100 purified water

(Formulation Example 5) Lotion Mass%
Baicalin 2.0
Baicalein 2.0
Glycerin 3.0
1,3-butylene glycol 5.0
Ethanol 15.0
Iron oxide 0.2
Zinc oxide 0.5
Kaolin 2.0
Trehalose 2.0
Dextrin 0.5
Photosensitive element 201 Appropriate amount Photosensitive element 401 Appropriate amount Fragrance Appropriate amount
Residue with 100 purified water

(Formulation Example 6) Lotion weight%
Sorbitol 2.0
1,3-butylene glycol 4.0
Rosmarinic acid 0.5
Polyoxyethylene (25) oleyl ether 2.0
Carboxyvinyl polymer 0.2
Ethanol 15.0
Purified water remaining

(Prescription Example 7) Ointment Mass%
Myricetin 2.0
l-Menthol 3.0
DL-Camphor 0.5
Propylene glycol 15.0
Carboxyvinyl polymer 1.0
Hydroxyethyl cellulose 0.1
Triethanolamine 0.7
Purified water 25.0
Ethanol 100 residue

(Prescription Example 8) Patch%
l-Menthol 3.0
DL-Camphor 1.5
Polyacrylic acid 4.5
Sodium polyacrylate 1.5
Sodium carboxymethylcellulose 4.0
Epigallocatechin gallate 1.2
Glycerin 20.0
Sorbitol 5.0
Polyoxyethylene nonylphenyl ether 0.5
Kaolin 5.0
Castor oil 1.0
Purified water remaining The above-mentioned components dissolved, dispersed and kneaded were spread on a polyester non-woven fabric so as to be 1000 g per 1 m ² to produce a patch.

(Prescription Example 9) Body cleaner Mass%
Epigallocatechin 1.5
Sodium citrate 2.0
Potassium laurate 15.0
Potassium myristate 5.0
Propylene glycol 5.0
Sorbitol 3.0
Polyethylene powder 0.5
Hydroxypropyl chitosan solution 0.5
Preservative appropriate amount
pH adjuster appropriate amount perfume appropriate amount
Residue with 100 purified water

(Formulation Example 10) Face wash mass%
Rosmarinic acid 2.8
Glyoxylic acid 0.5
Glyceryl monopalmitate 3.0
Polyoxyethylene behenyl ether 2.0
Octyldodecyl laurate 5.0
Lauryl hydroxysulfobetaine 1.0
Polyoxyalkylene-modified organopolysiloxane 4.0
Glycerin 8.0
Trehalose 2.0
2-phenoxyethanol 0.5
Perfume
Residue with 100 purified water

It is a figure which shows the production | generation pathway of a terminal glycation substance (AGEs). In the figure, HN-Lys represents the side chain amino residue of lysine. In the figure, H-X-Protein represents an arbitrary side chain of the amino acid constituting the protein.

Claims (3)

A preparation for degrading a protein-reducing sugar cross-linking substance, which is a Maillard reaction product, containing rosmarinic acid or a salt thereof as an active ingredient . The preparation according to claim 1, which is in a form for oral administration. The preparation according to claim 1, which is in a form for transdermal administration.

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