Sakura extract, AGEs decomposing agent and application thereof

Abstract

The present invention provides an oriental cherry flower extract containing 200 mg / kg or more of neochlorogenic acid as an active ingredient. The oriental cherry flower extract provided by the invention can promote the degradation of saccharification products, and can improve cell oxidative stress or inflammatory response caused by the saccharification products.

Description

Technical Field

[0001] This invention relates to the fields of plant extracts and AGEs decomposition agents. Background Technology

[0002] Advanced glycation end products (AGEs) are products of excessive sugar and protein binding. They can combine with and damage the body's tissues and cells, accelerating aging and leading to many chronic degenerative diseases such as diabetes, Alzheimer's disease, and atherosclerosis.

[0003] The formation of AGEs is generally considered to be mainly due to non-enzymatic glycosylation. That is, in the initial reaction, the amino groups in the protein react with the aldehyde groups of reducing sugars such as glucose to produce non-enzymatic glycosylation (Glycation), forming Amadori rearrangement products via Schiff base. In the later reaction, the above rearrangement products undergo long and complex ring-opening and condensation processes to form AGEs.

[0004] Inhibiting the formation of AGEs (Advanced Glycation End Products) can play a role in anti-aging and preventing various chronic degenerative diseases. Numerous research institutions have investigated methods to inhibit AGE formation. Among these studies, some have found that flavonoids in cherry blossoms have a significant inhibitory effect on AGE formation.

[0005] For example, Chinese patent application CN102883733 discloses a cherry blossom extract containing at least one compound selected from 1-O-(E)-caffeoyl-β-D-glucopyranoside, 1-O-(E)-coumaroyl-β-D-glucopyranoside, 1-O-(E)-cinnamyl-β-D-glucopyranoside, kaempferol 3-O-(6”-malonyl)-β-D-glucopyranoside, and quercetin 3-O-(6”-malonyl)-β-D-glucopyranoside as an active ingredient. It also discloses that this cherry blossom extract can effectively inhibit the formation of AGEs in fibroblasts.

[0006] Furthermore, Chinese patent application CN114794473 discloses a method for preparing cherry blossom powder with AGE-inhibiting activity. This method includes: S1 washing, S2 countercurrent extraction, S3 filtration, S4 concentration, S5 sterilization, S6 freeze-drying, and S7 sieving. In the S3 filtration step, a first filter bag of 38–180 μm, a second filter bag of 1–10 μm, a filter membrane of 100–200 nm, a first ultrafiltration membrane of 2–5 WDa, and a second ultrafiltration membrane of 1–5 kDa are used sequentially for filtration. In the S4 concentration step, a reverse osmosis membrane or nanofiltration membrane is used for concentration. The extract obtained by this method has a total flavonoid content exceeding 15%, and its main active components include 1-O-caffeoyl-β-D-glucopyranoside and quercetin 3-O-β-D-glucopyranoside, exhibiting significant AGEs inhibition.

[0007] However, none of the aforementioned prior art records that cherry blossom extract contains neochlorogenic acid, and the prior art focuses on the inhibitory effect of cherry blossom extract on AGEs formation, without studying how to decompose already formed AGEs. Summary of the Invention

[0008] The purpose of this invention is to provide an AGEs cherry blossom extract that can decompose already formed AGEs, and an AGEs decomposing agent containing the extract.

[0009] Through repeated research, this invention discovered that the neochlorogenic acid component in cherry blossom extract has the effect of degrading AGEs, and a cherry blossom extract with high neochlorogenic acid content and excellent AGEs degradation effect was prepared by a specific preparation method, thus completing this invention.

[0010] The present invention includes the following embodiments.

[0011] [1] A cherry blossom extract from Kanzan, which contains more than 200 mg / kg of neochlorogenic acid as an active ingredient.

[0012] [2] The cherry blossom extract of Kanzan described in [1] is a water extract.

[0013] [3] The cherry blossom extract of Guanshan as described in [1] or [2] also contains one or more of the following: astragaloside, rutin, chlorogenic acid, quercetin, protocatechuic acid, succinic acid, and quinic acid.

[0014] Use of any one of the following [4][1] to [3] in food, medicine or cosmetics.

[0015] [5] According to the uses described in [4], the food is in the form of an oral liquid, tablet, capsule or solid beverage.

[0016] [6] According to the uses described in [4], the medicine is a capsule, tablet, pill, granule, oral liquid, or injection.

[0017] [7] According to the use described in [6], the medicine contains any one of [1] to [3] cherry blossom extract and a pharmaceutically acceptable carrier.

[0018] Use of the Kanzan cherry blossom extract as described in any one of [8][1] to [3] in the preparation of a saccharification product decomposition agent.

[0019] Invention Effects

[0020] The cherry blossom extract of Guanshan in this invention can promote the degradation of glycation products and improve cellular oxidative stress or inflammatory response caused by glycation products. Attached Figure Description

[0021] Figure 1 : Standard curve of neochlorogenic acid concentration.

[0022] Figure 2 : Bar chart showing the chlorogenic acid content of the extracts of each embodiment and comparative example.

[0023] Figure 3 : A bar chart showing the AGEs decomposition rates of each embodiment and comparative example.

[0024] Figure 4 : A bar chart showing the CML decomposition rate of each embodiment and comparative example.

[0025] Figure 5 : A bar chart showing the CEL decomposition rate of each embodiment and comparative example.

[0026] Detailed Implementation of the Invention

[0027] The Kanzan cherry blossom extract of the present invention is prepared by a specific method, thereby containing a high proportion of neochlorogenic acid. By containing a high proportion of neochlorogenic acid in the Kanzan cherry blossom extract, excellent glycation product degradation function can be achieved.

[0028] It should be noted that the “glycation products” in this application include one or more of the following: advanced glycation end products (AGEs), human carboxymethyl lysine (CML), human carboxyethyl lysine (CEL), glyoxal (GO), methylglyoxal (MGO), 3-deoxyglucuronide (3-DG), or dimethylglyoxal.

[0029] Furthermore, the "glycation product degradation" mentioned in this application refers to the degradation of glycation products already generated in cells, which is different from the effect of inhibiting the generation of glycation products from the generation end.

[0030] The Kanzan cherry blossom extract of this application contains neochlorogenic acid at a concentration of 200 mg / kg or more as an active ingredient, preferably at a concentration of 300 mg / kg or more. By containing a high proportion of neochlorogenic acid, the Kanzan cherry blossom extract of this application can efficiently degrade already formed glycation products, thereby improving cellular oxidative stress or inflammatory responses caused by glycation products.

[0031] The extract of cherry blossoms from Kanzan in this application is preferably an aqueous extract, and its active ingredients preferably also contain one or more of the following: astragaloside, rutin, chlorogenic acid, quercetin, protocatechuic acid, succinic acid, and quinic acid.

[0032] The *Kanzan cherry blossom* extract of this application can be used in food, pharmaceuticals, cosmetics, and other applications. The food form may include, for example, oral liquid, tablets, capsules, or solid beverages. The pharmaceutical form may include, for example, capsules, tablets, pills, granules, oral liquids, or injections, and the pharmaceutical contains the *Kanzan cherry blossom* extract of this application and a pharmaceutically acceptable carrier.

[0033] The preparation method of the Kanzan cherry blossom extract of this application is described below.

[0034] (1) Extraction process

[0035] Dried Kanzan cherry blossoms or fresh dried cherry blossoms are crushed or ground, or moistened with water and then crushed or ground into a slurry. The resulting slurry is then added to an extractor, with the water-to-cherry blossom raw material mass ratio controlled at 5:1 to 20:1, the extraction temperature at 25 to 45°C, and the extraction time at 30 to 120 minutes to obtain an extract. The extractor can be a commonly used extractor, such as a multi-functional extraction tank.

[0036] (2) Filtration and Concentration Process

[0037] The extract obtained in the above extraction process is filtered through an 80-150 mesh filter to obtain a filtrate. The filtrate is then filtered through a 200-800 nm microfiltration membrane, such as a ceramic membrane. The resulting retentate is then concentrated using a high-temperature resistant reverse osmosis (RO) membrane. A Brix 5-20 concentrate is obtained. In this application, Brix 5-20 represents the content of soluble solids, and Brix 5-20 represents the refractive index determined by refractive index measurement.

[0038] (3) Drying process

[0039] The concentrate is freeze-dried or spray-dried to obtain a powdered product. Example

[0040] The present invention will be described in more detail below through embodiments. The embodiments are only used to illustrate the ideas and effects of the present invention, and the present invention is not limited to the configuration in the embodiments.

[0041] <Preparation of Cherry Blossom Extract>

[0042] Example 1

[0043] (1) Extraction process

[0044] The dried Kanzan cherry blossoms were crushed, moistened with water, and then added to a crusher to grind the cherry blossoms into a slurry. The resulting slurry was then added to a multi-functional extraction tank, and water was added at a mass ratio of 10:1 (water to cherry blossom raw material). Water extraction was carried out at 45°C for 60 minutes to obtain the extract.

[0045] (2) Filtration and Concentration Process

[0046] The extract obtained in the above extraction process is filtered through a 100-mesh filter to obtain a filtrate. The filtrate is then filtered through a 500nm microfiltration membrane (ceramic membrane) to obtain a retentate. This retentate is then concentrated through a high-temperature resistant reverse osmosis (RO) membrane (model: RO5-8040) to obtain a Brix 20 concentrate.

[0047] (3) Drying process

[0048] The concentrate was freeze-dried at -18 to -24°C to obtain a powdered extract.

[0049] Example 2

[0050] Except for using fresh cherry blossoms and directly blending them into a paste, the powdered extract was obtained in the same manner as in Example 1.

[0051] Comparative Example 1

[0052] In step (2), the extract is filtered through a filter screen only. A concentrated Brix 20 solid solution is obtained, otherwise a powdered extract is obtained in the same manner as in Example 1.

[0053] Comparative Example 2

[0054] In step (2), the extract is filtered through a microfiltration membrane to obtain a concentrated Brix20 solid solution. Otherwise, a powdered extract is obtained in the same manner as in Example 1.

[0055] Comparative Example 3

[0056] In step (2), the extract is concentrated only by passing it through a reverse osmosis (RO) membrane to obtain Brix 20 concentrate in solid form. Otherwise, the powdered extract is obtained in the same manner as in Example 1.

[0057] Comparative Example 4

[0058] In step (2), the extract is filtered through a filter screen, and the resulting retentate is concentrated through a reverse osmosis (RO) membrane to obtain Brix 20 concentrate. Otherwise, a powdered extract is obtained in the same manner as in Example 1.

[0059] Comparative Example 5

[0060] In step (2), the extract is filtered through a microfiltration membrane, and the resulting retentate is concentrated through a reverse osmosis (RO) membrane to obtain Brix 20 concentrate. Otherwise, a powdered extract is obtained in the same manner as in Example 1.

[0061] Comparative Example 6

[0062] Cherry blossom powder prepared according to the method in Example 1 of CN114794473.

[0063] In Example 1, the extract was first filtered through a 150μm first filter bag. The filtrate then flowed through a closed pipe into a pipeline filter containing a 5μm second filter bag, followed by filtration through a ceramic membrane device (model KM1030-CM-122) with a built-in 200nm ceramic filter membrane. Next, it was filtered through an ultrafiltration membrane device (model KM8040-NF-G6) with a built-in 5WDa first ultrafiltration membrane, and finally through an ultrafiltration membrane device (model KM8040-NF-G6) with a built-in 3KDa second ultrafiltration membrane to obtain a retentate. This retentate was then concentrated using a reverse osmosis membrane device (model KM8040-NF-G16) to a relative density of 1.07, obtaining a concentrated solution. After sterilization, the solution was transferred to trays via a distributor and freeze-dried in a freeze dryer to obtain cherry blossom powder.

[0064] Comparative Example 7

[0065] Cherry blossom powder prepared according to the method of the embodiment in CN102883773.

[0066] The flowers of cherry blossom [Mountain Cherry (Sekiyama), Prunus lannesiana Wils. cv. Sekiyama] were extracted with 30% (w / w) aqueous ethanol for 1 hour (60°C). Subsequently, the extract was filtered through a 150 μm filter bag to obtain a Brix 20 concentrate of solids, yielding the cherry blossom extract (yield 5 wt%).

[0067] <Determination of Chlorogenic Acid Content>

[0068] The chlorogenic acid content of the extracts in the examples and comparative examples was determined by the standard curve method.

[0069] Standard curve plotting

[0070] Neochlorogenic acid standards of different concentrations were prepared, and the chromatograms of neochlorogenic acid in each standard were determined by liquid chromatography. The peak area of ​​neochlorogenic acid was calculated, and a standard curve was plotted with the standard concentration as the x-axis and the peak area as the y-axis. Figure 1 As shown.

[0071] Determination of chlorogenic acid content

[0072] A certain amount of the extracts from the examples and comparative examples were prepared into solutions. The chromatograms of each solution sample were then determined by liquid chromatography, and the peak area corresponding to neochlorogenic acid was calculated. The corresponding neochlorogenic acid concentration was then found in the standard curve using this peak area. Finally, the neochlorogenic acid concentration was divided by the concentration of the extract solution sample to obtain the neochlorogenic acid content in the extract.

[0073] The content of neochlorogenic acid in the extracts of Examples 1-2 and Comparative Examples 1-7 is as follows: Figure 2 As shown.

[0074] <Degradation Test of Glycoproducts>

[0075] Experimental procedures for the degradation of AGEs and glycation products

[0076] HSF cells were administered at a dose of 1×10 5 Cells were seeded at a density of 1 / mL in 6-well cell culture plates and incubated at 37°C and 5% CO2 for 6 h. Then, 0.4 mM acetone aldehyde solution was added to induce glycosylation. After 24 hours of culture, 150 μg / mL of cherry blossom extract was added to each sample. After another 24 h of culture, the degradation rates of AGEs and other glycation products were measured. The content of AGEs was determined according to the ELISA kit method for human advanced glycation end products (AGEs), human carboxymethyl lysine (CML), and human carboxyethyl lysine (CEL). The degradation rates of AGEs, CML, and CEL in each group were calculated using the following formula:

[0077] Glycosylation product degradation rate = (concentration of glycosylation products in the untreated group - concentration of glycosylation products in the extract-treated group) / concentration of glycosylation products in the untreated group

[0078] The AGEs decomposition rate, CML decomposition rate, and CEL decomposition rate of the extracts from Examples 1-2 and Comparative Examples 1-7 are as follows: Figures 3-5 As shown.

[0079] Depend on Figures 2-5 It can be seen that the chlorogenic acid content of the extracts obtained in Comparative Examples 1-5 (which had fewer filtration steps than those in the present invention) and Comparative Example 7 (which had more filtration steps than those in the present invention) was lower than that of the extracts in the present invention.

[0080] Furthermore, the AGEs decomposition rate, CML decomposition rate, and CEL decomposition rate of the extracts in the embodiments of this application are all higher than those in Comparative Examples 1-7. The decomposition rate of saccharified products is significantly correlated with the neochlorogenic acid content in the cherry blossom extract.

[0081] Therefore, the extract of this application can achieve excellent decomposition effect of glycosylated products.

Claims

1. A cherry blossom extract from Kanzan, which contains more than 200 mg / kg of neochlorogenic acid as an active ingredient.

2. The Kanzan cherry blossom extract according to claim 1 is an aqueous extract.

3. The cherry blossom extract of Guanshan according to claim 1 or 2, further comprising one or more of astragaloside, rutin, chlorogenic acid, quercetin, protocatechuic acid, succinic acid, and quinic acid.

4. Use of the Kanzan cherry blossom extract according to any one of claims 1 to 3 in food, medicine or cosmetics.

5. The use according to claim 4, wherein the food is in the form of an oral liquid, tablet, capsule or solid beverage.

6. According to the use described in claim 4, the medicine is a capsule, tablet, pill, granule, oral liquid, or injection.

7. The use according to claim 6, wherein the pharmaceutical product contains the Kanzan cherry blossom extract as described in any one of claims 1 to 3 and a pharmaceutically acceptable carrier.

8. Use of the Kanzan cherry blossom extract according to any one of claims 1 to 3 in the preparation of a saccharification product decomposition agent.

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