JP6385097B2 - Cutting agent - Google Patents

Description

  The present invention relates to a decomposition agent (cleaving agent) having an excellent action of decomposing (cleaving) “AGEs” which are products produced as a result of the Maillard reaction. Here, “AGEs” means “advanced glycation end products”.

(Maillard reaction inhibitor and cutting agent)
-1-
As is well known, the Maillard reaction is a non-enzymatic reaction that produces a brown substance when a reducing sugar and an amino compound (amino acid, peptide, protein, etc.) are heated. .
-2-
As this reaction progresses, it is said that the quality of foods will decrease. Therefore, in order to suppress the deterioration of food quality, it is desired to develop a substance (that is, “inhibitor”) that effectively suppresses the progress of the Maillard reaction.
-3-
Maillard reaction also occurs in vivo, which may contribute to aging and diabetic complications. Therefore, also from this viewpoint, development of an effective and safe Maillard reaction inhibitor is desired.
-4-
In addition, since it is difficult to sufficiently suppress the occurrence of the Maillard reaction product in the living body, a “decomposing agent (that is, a cutting agent)” that effectively decomposes (cuts) the Maillard reaction product that has occurred. It is also as important as the development of Maillard reaction inhibitors.
-5-
The present invention relates to the latter decomposition agent (cutting agent) among the above-described inhibitors and decomposition agents (cutting agents).
The term “decomposing agent” and “cleaving agent” in the latter are synonymous, but since the evaluation is often performed based on the degree of “cutting rate or cutting activity”, the term “cutting agent” is mainly used in this specification. To decide to use,

(Patent Document 1)
-1-
Claim 1 (and paragraph 0009) of Japanese Patent Application Laid-Open No. 2007-119373 (Patent Document 1, filed on October 26, 2005) includes a Maillard reaction product containing an active substance selected from various plant extracts. Degradants (ie, cutting agents) are shown.
Claim 2 shows that the active substance is an extract obtained by extracting plant-derived tissue with water or hot water.
The first to fourth aspects relate to the definition of the cleavage activity of the above-described decomposition agent (active substance).
The fifth aspect relates to a food or drink containing the Maillard reaction product decomposing agent.

-2-
In paragraph 0019, it is described that the above-mentioned active substance may be extracted from various parts of a plant as it is or after pulverization, or may be extracted as it is or after pulverization with a solvent. There is.
In addition, paragraph 0019 describes that examples of the solvent include water, hot water, alcohol, ketone, ether, nitrile, ester, aromatic solvent, halogenated solvent and the like.

-3-
The paragraph 0021 describes the food and drink according to claim 5 (and supplements, pharmaceuticals, cosmetics and other products containing the Maillard reaction product degradation agent), and paragraphs 0039 to 0048 include a plant system. A large number of additives derived from raw materials, inorganic raw materials and animal raw materials are exemplified.

-4-
In the paragraph 0026, there is a description that the above-mentioned food, drink, cosmetics or pharmaceuticals may appropriately contain the components and additives exemplified below as necessary.
Paragraph 0059 mentions propyl gallate as an example of an antioxidant.
Paragraph 0075 mentions gallic acid as an example of a dye agent. However, it describes the possibility of use as a dye agent. (Incidentally, gallic acid turns black in combination with iron ions, as described later in Patent Document 3, and is used as a hair dye.)
The paragraph 0076 mentions rose essential oil as an example of a fragrance.
In paragraph 0999, there is a description that “Table 1 and Table 2 show the carbon-carbon bond cleavage activity (α-dicarbonyl compound decomposition activity) of the oxal group in the control example and Examples 1 to 47”. This states that "AGE-protein crosslinker model compound has an α-dicarbonyl structure".

-5-
In paragraphs 0083 to 0098 according to the examples and comparative examples, the green tea leaf extract is concentrated and then subjected to column chromatography to collect a fraction in which catechins (catechin, epigallocatechin, epigallocatechin gallate) are eluted. There is a description about the result when cutting activity was calculated | required using what was made as an extract or powder.
In Tables 1 and 2 of paragraphs 0100 to 0101, “relative ratio (%)” when various plant extracts are used and “relative” when the reference PTB is 100%. "Cleavage activity" is shown, and there are 17 of 47 cases with relative cleavage activity values exceeding 100, including 694 and 456, in comparison with the control PTB.

-6
Incidentally, in Patent Document 2 (filed four years and eight months before Patent Document 1) described below, “100 mM PTB” is used as a positive control as described in the paragraph 0115. And when the restoration rate of the control (protein cleavage crosslinking activity according to paragraph 0114) is 100%, the restoration rate of Examples using various plant extracts is 22 to 90%. There is no naturalness.

−7−
Then, returning to this Patent Document 1 and looking at the description of the control PTB, paragraph 0088 includes “10 mM PTB solution” as the reaction solution composition.
That is, in Patent Document 1, since the PTB that is a control is a PTB having a concentration of “10 mM” that is 1/10 of the concentration (100 mM) of Patent Document 2, the tables in paragraphs 0100 to 0101 of Patent Document 1 It is natural that the cleavage rate of Examples 1 and 2 is higher compared to the positive control.

-8-
In addition, in paragraph 0104 according to Comparative Examples 12 to 38 of Patent Document 1, the 27 plant extracts listed in Claim 1 of Patent Document 2 and its paragraph 0123 according to the application of another company preceding that are exactly the same. They are listed as they are, and their degradation activity measurement results are the same as those of Patent Document 2 (90 for the highest pomegranate extract and 22 for the lowest Genkosho extract).
The fact that the same results as in Patent Document 2 were obtained means that the concentration of the control PTB at that time is not 100 mM, but the wrinkles do not match, but as described above, in Patent Document 1, as in paragraph 0088 Since “10 mM PTB solution” is used as a positive control, the description in paragraph 0104 of Patent Document 1 seems to be contradictory.

(Patent Document 2)
-1-
Claim 1 of JP-A-2002-241299 (Patent Document 2, filing date: February 13, 2001) includes at least one plant extract selected from 27 types of plants (including tea tree and rose) Maillard reaction repair agents containing are shown.
In claim 2, the Maillard reaction repair agent according to claim 1 having protein cross-linking cleavage activity is shown.
Paragraph 0032 mentions damask rose as an example of the above “rose”.
In paragraph 0062 there is a reference to Querx Infectria.
That paragraph 0081 mentions propyl gallate as an example of an antioxidant.
Paragraph 0097 mentions gallic acid as an example of a dye agent.
The paragraph 0098 mentions rose essential oil as an example of a fragrance.

-2-
In Production Examples 1 to 6 according to the examples, various plants (also referred to as roses) are sequentially arranged in the order of “water; 50% ethanol solution; 50% 1,3-butylene glycol solution; 50% 1,3-butylene. It is shown that the protein cross-linking cleavage activity (repair rate) was measured using an extract extracted by immersion in a “glycol solution; 50% propylene glycol solution; water”.

-3-
The results at that time are shown in Table 1 of the paragraph 0117. When the restoration rate of the control (100 mM PTB) is 100%, the restoration rate when the above-mentioned various plant extracts are used is 22 to 90. %. In addition, the restoration rate of chanoki is 30%, and the restoration rate of roses is 74%.
According to the description of paragraph 0118 related to Table 1, “each plant extract was confirmed to have significant protein cross-linking cleavage activity (22-74%) compared to blank,” Because the repair rate of the control (100 mM PTB) is 100%, there is no case where the repair rate (22-74%) of each sample exceeds the repair rate of the control.

(Patent Document 3)
-1-
Claim 1 of Japanese Patent Application Laid-Open No. 2014-009160 (Patent Document 3) includes “at least one (a) selected from the group consisting of tannic acid, gallic acid, and gallic acid derivatives; and an ammonium salt (b). The hair dye composition containing the 1st agent (1) containing "and the 2nd agent (2) containing an iron salt" is shown.
-2-
However, the “hair dye” according to Patent Document 3 (the hair dye for the purpose of fixing the pigment to the hair by coloring with the reaction of the first agent and the second agent (iron salt, etc.)) of the present invention. It is unrelated to the target “Maillard reaction product degradation agent (that is, cleavage agent) having an excellent cleavage activity of the AGE-protein crosslinker model”.

JP 2007-119373 A JP 2002-241299 A JP 2014-009160 A

(Regarding Patent Document 1)
-1-
In paragraph 0026 of Patent Document 1 (the filing date is October 26, 2005), “In addition, the food, beverage, cosmetics or pharmaceuticals of the present invention are exemplified in the following (1) to (45) as necessary. It may contain components and additives as appropriate ". Then, in accordance with the description, in paragraph 0075, 57 dyes are listed as “(40) dye agent”, and 53rd is “gallic acid”.
-2-
However, in the section of [Example] after paragraph 0088, 85 examples of Examples 1 to 47, Comparative Examples 1 to 11, and Comparative Examples 12 to 38 are listed, but “dye agent” is added. There are no examples.
-3-
If it is recognized that a certain kind of dye has an effect as a Maillard reaction product decomposition agent aimed by Patent Document 1, it is described as an active ingredient in claim 1 instead of “(40) Dye agent”. Should have been.

(Regarding Patent Document 2)
-1-
In paragraph 0045 of Patent Document 2 (the filing date is February 13, 2001), “In addition to the above-mentioned essential components, the components exemplified below may be produced by arbitrarily selecting and using additives in combination. The content in the preparation is not particularly specified, but is usually preferably 0.0001 to 50%. "Hereinafter, the additives (1) to (45) are mentioned. The paragraph (0097) “(40) Dye Agent” contains 57 dyes, 53 of which are “gallic acid”.
However, in the [Example] section after paragraph 0107, there is no example in which “dye agent” is added.
-2-
As in the case of Patent Document 1, if it is recognized that a certain type of dye has an effect as a Maillard reaction product decomposition agent aimed by Patent Document 2, it is not an item of “(40) Dye Agent”. And should be described as an active ingredient in claim 1.
-3-
Note that the description of “(40) dye agent” in paragraph 0097 of Patent Document 2 is the same as the description of “(40) dye agent” in paragraph 0075 of Patent Document 1 described above. is there. (Since Patent Document 1 is related to an application that is four years, eight months and a half later than Patent Document 2, the description of Patent Document 1 is the same as the description of Patent Document 2.)

(Regarding Patent Document 3)
The “hair dye” according to Patent Document 3 is for the purpose of fixing the pigment to the hair by developing a color by the reaction of the first agent and the second agent. It is not related to the Maillard reaction product decomposition agent (that is, the cleaving agent) having an excellent cleaving activity in the product model.

(First cutting agent of the present invention)
-1-
The first cleaving agent of the present invention is characterized by comprising free gallic acid as an active ingredient when the Maillard reaction product decomposing agent is referred to as “cleaving agent”.
-2-
Here, free gallic acid means 3,4,5-trihydroxybenzoic acid (3,4,5-trihydroxybenzoic acid), and the effect of being free is as a “cleaving agent”. Demonstrate. ("Free type" means that it is not a gallic acid derivative as described below. Depending on the environmental pH, the COOH moiety may be in an ionic or salt form such as COO-. Is also included in the free form.)
Incidentally, gallic acid derivatives such as alkyl gallate and tannin are naturally not included in the category of free gallic acid.

(Second cutting agent of the present invention)
-1-
When the second cleavage agent of the present invention refers to the Maillard reaction product decomposition agent as a “cleavage agent”,
(A) The cutting agent is composed of a gallic acid-containing composition containing a plant-derived free gallic acid, and
(I) When a plant is carbonized under low temperature and reduced pressure conditions using a carbonization device equipped with a vacuum carbonization mechanism, the dry solid residue remaining in the carbonization device after the low temperature vacuum carbonization operation is cryogenically vacuum distilled. When the residue R is called a residue extract E and the solvent extract from the low temperature vacuum distillation residue R is called residue extract E,
At least a part of the gallic acid-containing composition is a plant-derived gallic acid-containing composition comprising the low-temperature vacuum distillation residue R or / and the residue extract E,
It is characterized by.
-2-
Various kinds of plants can be used as long as free gallic acid is contained on the residue side by low-temperature vacuum distillation operation. An example of a typical (or noteworthy) plant is a rose, in particular a damask rose.
-3-
The low temperature vacuum distillation operation is (A) the pressure is, for example, a reduced pressure condition of 100 mmHg or less (more preferably 50 mmHg or less), and (B) the temperature is, for example, a low temperature condition of about 90 ° C. to room temperature. (C) About the moisture content of the plant to be subjected to dry distillation, for example, it is about the natural moisture content for raw plants, and about the moisture content of the frozen product for frozen storage plants.

-1-
(First cutting agent)
In the first cleaving agent of the present invention, free gallic acid as a compound exhibits an excellent effect as a “cleaving agent”. This fact is believed to have never been known before.

-2-
(Second cutting agent)
The 2nd cutting agent of this invention consists of a gallic-acid containing composition containing the plant-derived free gallic acid, and also paid its attention to the acquisition source and acquisition means of the target in addition to the target. Is.
The gallic acid-containing composition containing free gallic acid at this time is obtained through a step of carbonizing a plant under a low temperature and reduced pressure using a dry distillation apparatus equipped with a vacuum distillation mechanism.
This route is not only industrial, but also satisfies the safety and security of being derived from plants. In addition to the direct target, free gallic acid, Roses can be selected and used), so that the target product can be obtained in a form that includes other components contained in it and its decomposition products. It is also preferable in that it can be commercialized in accordance with the above.

  Next, the present invention will be further described with reference to examples and reference examples.

  The actual test is based on the "first test" using gallic acid and its derivatives, the "low-temperature vacuum distillation residue R" obtained by low-temperature vacuum distillation of plants (using roses), and the low-temperature vacuum distillation residue R The “secondary test” using “residue extract E”, which is an extract of the above solvent, is performed in a mixed manner. In this example, first, in the case of the “first test” Next, the case of the “secondary test” will be described.

[First test (part 1)]
(1) Reagent used for test As a reagent, the following five types of compounds were prepared.
1: Gallic acid (purity is 98.0% or more)
2: Ethyl gallate (purity is 98.0% or more)
3: Dodecyl gallate (purity unknown)
4: 3,4,5-trimethoxybenzoic acid (purity is 97.0% or more)
5: Syringic acid (purity unknown)
-Remarks-
・ Gallic acid is 3,4,5-trihydroxybenzoic acid ・ Alkyl gallate (ethyl, dodecyl) is a gallic acid whose COOH group is COOR (alkyl ester) ・ 3,4,5-trimethoxybenzoic acid Acid is one in which three OH groups of gallic acid are changed to OCH3 group (methoxy group). Syringic acid is gallic acid-3,5-dimethyl ether (that is, 4-oxy-3,5-dimethoxybenzoic acid).

(2) Method The cleavage activity of the AGE-protein crosslinker model was measured according to the method of Vasaran et al. (Nature, Vol. 382, p. 275-278, 1996).
That is, a reaction solution having the composition shown below was prepared in a 1.5 mL plastic tube and shaken at 37 ° C. for 4 hours).

“Reaction solution composition and addition amount”
-500 mM disodium hydrogen phosphate (pH 7.4) ... 800 μL
・ 100 mM 1-phenyl-1,2-propanedione: 100 μL
・ Sample solution …………………………… 100μL

"Measure cutting rate"
After completion of the reaction, 200 μL of 2N HCl was added and stirred to stop the reaction.
The liquid was filtered through a 0.2 μm filter to obtain an HPLC analysis sample solution.

`` HPLC analysis conditions ''
Column: YMC-Pack ODSA-312 150 × 6 mmI. D.
Eluent: 40% MeOH / 0.1% trifluoroacetic acid (TFA)
-Flow rate: 1.0 mL / min.
-Column temperature: 40 ° C
・ Detector: UV wavelength 273nm
・ Injection volume: 20μL
・ Retention time: about 12 minutes

"How to determine the cutting rate"
-1-
Since it can be assumed that 10 mM benzoic acid is liberated when all 1-phenyl-1,2-propanedione is cleaved, the cleavage rate was calculated according to the following equation.
Cleavage rate (%) = 100 × (peak area of benzoic acid generated from each analysis sample / peak area of 10 mM benzoic acid)
-2-
As a reminder, the concentration of benzoic acid at the above cleavage rate is annotated.
When 1-phenyl-1.2-propanedione is cleaved, 1-phenyl-1.2-propanedione and an equimolar amount of benzoic acid are produced, and the resulting benzoic acid is quantified by HPLC.
As in the above “HPLC analysis conditions”, the total amount of the reaction solution is 1 mL, and 100 μL of 100 mM 1-phenyl-1.2-propanedione is added to the reaction solution. .2-Propanedione concentration is 10 mM. Therefore, when the total amount of 1-phenyl-1.2-propanedione is decomposed, the benzoic acid concentration in the reaction solution becomes 10 mM.

(2) Measurement result of cutting rate The measurement result of the cutting rate for the sample solution prepared above is shown in Table 1 below.

(3) Discussion The cleavage activity of the AGE-protein cross-linking product model was as follows when the compound amount was 100 mM.
1: Gallic acid had a strong activity almost equal to that of the positive control.
2: The cleavage activity of ethyl gallate was about 1/3 of the positive control.
3: The cleavage rates in the case of dodecyl gallate, 3,4,5-trimethoxybenzoic acid, and syringic acid were all very low compared to the positive control.

[First test (2)]
(1) Reagent used in the test Since the tendency of the cleavage rate for gallic acid and a compound similar to gallic acid was grasped in the first test (part 1), the reagent was used in the first test (part 1). Cleavage activity was examined by focusing only on “gallic acid” for which favorable results were obtained, and at two concentrations of 100 mM and 10 mM.

(2) Measurement result of cutting rate Table 2 shows the results.

(3) Consideration-1-
When the amount of gallic acid was 100 mM (Example 2), the cleavage activity of the AGE-protein cross-linker model was as strong as the positive control.
-2-
When the amount of gallic acid was 10 mM (Example 3), the cleavage rate was about 1/3, but in Example 3, the amount of gallic acid was 1/10 than in Example 2. Therefore, it can be seen that the cutting rate exceeding the expectation or expectation was obtained.

[Third] Secondary test-1-
Rose (damask rose), which is a raw material plant, was dry-distilled under low-temperature decompression conditions of 40 ° C. × 20 mmHg. And the residue R of the dry state which remains in a carbonization apparatus after the low temperature vacuum distillation process was acquired.
Moreover, the residue extract E was acquired by extracting a part of the residue R using 50% ethanol.
The low-temperature vacuum distillation residue R includes various components, and the residue extract E includes various components, both of which include gallic acid. It was confirmed that
And it has been qualitatively confirmed that both of these low-temperature reduced-pressure dry distillation residues R and the residue extract E have cutting characteristics.

If the cutting agent of the present invention (that is, Maillard reaction product decomposition agent) is used, a reliable and preferable cutting rate is achieved.
Therefore, the cutting agent of the present invention is useful as cosmetics, foods, supplements, pharmaceuticals, etc., or as additives added to them. In particular, it is optimal as an effective supplement for anti-aging and anti-lifestyle diseases.

Claims (1)

When the Maillard reaction product decomposition agent is referred to as a “cleaving agent”,
(A) The cutting agent is composed of a gallic acid-containing composition containing a plant-derived free gallic acid, and
(I) When a plant is carbonized under low temperature and reduced pressure conditions using a carbonization device equipped with a vacuum carbonization mechanism, the dry solid residue remaining in the carbonization device after the low temperature vacuum carbonization operation is cryogenically vacuum distilled. When the residue R is called a residue extract E and the solvent extract from the low temperature vacuum distillation residue R is called residue extract E,
At least a part of the gallic acid-containing composition is a plant-derived gallic acid-containing composition comprising the low-temperature vacuum distillation residue R or / and the residue extract E,
The plant is damask rose ,
A cutting agent characterized by.