CN114939120A - Sialic acid composition and application thereof in relieving inflammation - Google Patents
Sialic acid composition and application thereof in relieving inflammation Download PDFInfo
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- CN114939120A CN114939120A CN202210518150.6A CN202210518150A CN114939120A CN 114939120 A CN114939120 A CN 114939120A CN 202210518150 A CN202210518150 A CN 202210518150A CN 114939120 A CN114939120 A CN 114939120A
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- sialic acid
- trehalose
- acid composition
- composition
- inflammatory
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- SQVRNKJHWKZAKO-OQPLDHBCSA-N sialic acid Chemical compound CC(=O)N[C@@H]1[C@@H](O)C[C@@](O)(C(O)=O)OC1[C@H](O)[C@H](O)CO SQVRNKJHWKZAKO-OQPLDHBCSA-N 0.000 title claims abstract description 64
- 239000000203 mixture Substances 0.000 title claims abstract description 36
- 206010061218 Inflammation Diseases 0.000 title claims abstract description 11
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- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 claims abstract description 21
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- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 claims abstract description 21
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Images
Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7016—Disaccharides, e.g. lactose, lactulose
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
- A61K31/19—Carboxylic acids, e.g. valproic acid
- A61K31/195—Carboxylic acids, e.g. valproic acid having an amino group
- A61K31/197—Carboxylic acids, e.g. valproic acid having an amino group the amino and the carboxyl groups being attached to the same acyclic carbon chain, e.g. gamma-aminobutyric acid [GABA], beta-alanine, epsilon-aminocaproic acid or pantothenic acid
- A61K31/198—Alpha-amino acids, e.g. alanine or edetic acid [EDTA]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7012—Compounds having a free or esterified carboxyl group attached, directly or through a carbon chain, to a carbon atom of the saccharide radical, e.g. glucuronic acid, neuraminic acid
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/40—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing nitrogen
- A61K8/44—Aminocarboxylic acids or derivatives thereof, e.g. aminocarboxylic acids containing sulfur; Salts; Esters or N-acylated derivatives thereof
- A61K8/442—Aminocarboxylic acids or derivatives thereof, e.g. aminocarboxylic acids containing sulfur; Salts; Esters or N-acylated derivatives thereof substituted by amido group(s)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/60—Sugars; Derivatives thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q11/00—Preparations for care of the teeth, of the oral cavity or of dentures; Dentifrices, e.g. toothpastes; Mouth rinses
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
- A61K2800/59—Mixtures
- A61K2800/592—Mixtures of compounds complementing their respective functions
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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Abstract
The invention relates to the technical field of sialic acid functional products, in particular to a sialic acid composition and application thereof in relieving inflammation. The sialic acid composition comprises: sialic acid, trehalose, and arginine; the mass ratio of the sialic acid to the trehalose is 1: (8-10); the pH value of the water solution of the composition is 6-8. According to the invention, sialic acid has a certain anti-inflammatory effect, and a composition compounded by sialic acid, trehalose and arginine is provided on the basis of the sialic acid, so that the anti-inflammatory effect is remarkably improved by compounding the sialic acid, the trehalose and the arginine, and the composition has an important significance in the field of anti-inflammatory application, especially in the field of alleviating stomatitis.
Description
Technical Field
The invention relates to the technical field of sialic acid functional products, in particular to a sialic acid composition and application thereof in relieving inflammation.
Background
Sialic acid (Sialic acid) is a derivative of a 9-carbon monosaccharide, also known as Sialic acid, a naturally occurring carbohydrate. Sialic acid is considered to be the transmitter of gangliosides and is one of the constituents of the brain, usually in the form of oligosaccharides, glycolipids or glycoproteins.
The existing research shows that sialic acid plays a very important role in the generation and development of the brain and nervous system, and is important for the normal development of brain functions of children. In addition, sialic acid is widely used in various fields of life, for example, in various kinds of foods, cosmetics, and health products.
However, studies on the function of sialic acid are still insufficient, and their use in a wider range is still under study.
Disclosure of Invention
In order to solve the problems in the prior art, the invention aims to provide a sialic acid composition and application thereof in relieving inflammation, wherein the anti-inflammatory capability of sialic acid is obviously improved by compounding sialic acid, trehalose and arginine.
In a first aspect, the present invention provides a sialic acid composition comprising: sialic acid, trehalose and arginine; the mass ratio of the sialic acid to the trehalose is 1: (8-10); the pH value of the water solution of the composition is 6-8.
According to the invention, sialic acid has a certain anti-inflammatory effect, and on the basis of the research, the process of exploring more functions of sialic acid is unexpectedly found that the anti-inflammatory capability of sialic acid is remarkably improved after sialic acid, trehalose and arginine are compounded. The specific combination mode is that the dosage of the sialic acid and the trehalose is within a certain mass ratio range, and then arginine is added to enable the pH value to reach 5-8, so that the composition is obtained.
Furthermore, in the sialic acid composition, the mass percentage of the sialic acid is more than or equal to 9%.
Further, the food comprises, by weight, 5-15 parts of sialic acid, 80-100 parts of trehalose and 1-10 parts of arginine.
Further, the beverage comprises, by weight, 9-11 parts of sialic acid, 80-90 parts of trehalose and 5-6.5 parts of arginine.
The invention further provides the application of the sialic acid composition in relieving inflammation.
The invention further provides application of the sialic acid composition in preparing a medicament for relieving inflammation or a cosmetic with a relieving effect.
In a second aspect, the invention provides an anti-inflammatory product comprising said sialic acid composition.
Further, the anti-inflammatory product is a cosmetic with soothing effect, or a pharmaceutical preparation with anti-inflammatory function. As a specific cosmetic category, the soothing cosmetics include toothpaste having an anti-inflammatory function.
Further, the method also comprises the following steps: any one or more of stabilizers, solubilizers, buffers, preservatives, thickeners, abrasives, humectants, flavoring agents, buffers, or other excipients.
Furthermore, the dosage form of the anti-inflammatory product is one or more of tablets, powders, granules, capsules, suppositories, aqueous solutions, ointments, creams, gels, inhalants, sprays, membranes, suspensions, troches, elixirs, syrups, water-in-oil or oil-in-water emulsions, oral liquids or dropping pills.
The invention has the following beneficial effects:
according to the research of the invention, sialic acid has a certain anti-inflammatory effect, but the anti-inflammatory effect is limited, trehalose and arginine are further compounded on the basis of the sialic acid, the anti-inflammatory effect of the obtained sialic acid composition is obviously improved, and compared with an untreated inflammatory cell model, the expression level of PGE2 is reduced by more than 35%, so that the sialic acid composition has an important significance in the field of inflammation alleviation. Particularly, the invention establishes an inflammatory reaction model on human gingival fibroblasts, and proves that the composition has a remarkable anti-inflammatory effect on the model, which indicates that the composition can be more specifically applied to cosmetics or sanitary products such as toothpaste, mouthwash and the like.
Drawings
FIG. 1 is a schematic diagram of MTT assay results of sialic acid provided in example 1 of the present invention.
Fig. 2 is a schematic diagram of MTT experimental results of the composition NANAPRO provided in example 2 of the present invention.
Detailed Description
The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention.
The reagents mentioned in the following examples are commercially available unless otherwise specified.
The test methods described in the following examples can be carried out according to a method conventional in the art unless otherwise specified.
Example 1
This example establishes an inflammatory response model based on IL-1 α stimulation of human gingival fibroblasts to study the anti-inflammatory effects of sialic acid as follows:
1. experimental Material
The cells used in the present invention are human gingival fibroblasts, batch No. 210708, commercially available. An inflammatory response model is obtained by induction with IL-1 alpha as a proinflammatory factor, the induction method can refer to the method described in the CN111139219A patent, and the anti-inflammatory effect of the composition can be effectively judged by detecting the content of PGE 2.
2. Experimental reagent
The present invention relates to reagents such as high-glucose dmem (gibco), fetal bovine serum (bright Lanzhou), PBS (doctor's Ded), mtt (Sigma), dmso (Sigma), dexamethasone (Sigma), and PGE2 ELISA kit (Abcam).
3. Testing instrument
The invention relates to CO 2 Incubator (Thermo, 150I), clean bench (Suzhou Antai, SW-CJ-1F), microplate reader (BioTek, Epoch).
4. Test method
(1) According to 4X 10 4 Cell/well seeding Density cells were plated onto 24-well plates, incubators (37 ℃, 5% CO) 2 ) And incubated overnight.
(2) The solutions of each experimental and control group were prepared according to the protocol shown in the following table, wherein the MTT experiment (shown in fig. 1) showed that the sample N-acetylneuraminic acid (cubilose, cubilotin, sialic acid) was based on gingival fibroblasts and showed no significant cytotoxicity in the concentration range of 6.25 mg/mL.
TABLE 1 protocol for the configuration of the experimental and control groups
(3) Administration and stimulation
According to the configuration scheme of table 1, when the cell plating rate in a 24-well plate reaches 40% -50%, the administration amount per well is 1mL, and each group is provided with 3 multiple wells. Then in an incubator (37 ℃, 5% CO) 2 ) The medium was continued for 24 h.
(4) Collecting cell supernatant
After 24h of culture, the cell culture supernatant was collected in an EP tube and stored frozen at-80 ℃ in a refrigerator.
(5) ELISA detection
Detection was performed according to the instructions for the PGE2 ELISA kit.
5. Result statistics and analysis
The results are expressed as Mean ± SD using GraphPad Prism mapping. The comparison between groups was performed by statistical analysis of t-test. Statistical analysis was two-tailed. P <0.05 was considered to have significant differences, and P <0.01 was considered to have very significant differences.
6. The result of the detection
The present invention gives the results shown in the following table:
TABLE 2 summary of PGE2 assay results for the experimental and control groups
Remarking: when the statistical analysis is carried out by using the t-test method, compared with the BC group, the significance is represented by #, the P-value <0.05 is represented by #, and the P-value <0.01 is represented by # #; significance was expressed as x, P-value <0.05 and P-value <0.01 compared to NC group.
The result shows that compared with the BC group, the secretion level of PGE2 in the NC group is extremely obviously increased (P <0.01), which indicates that the IL-1 alpha stimulation can extremely obviously improve the secretion amount of PGE2, and indicates that the IL-1 alpha stimulation condition is effective in the test.
Compared with the NC group, the PGE2 secretion level of the PC group was extremely significantly decreased (P <0.01), indicating that the positive control of this test was effective.
Compared with the NC group, the sample of the cubilose acid (cubilose acid, cubilotin and sialic acid) -6.25mg/mL can obviously inhibit the secretion of PGE2 caused by IL-1 alpha stimulation.
Example 2
In this example, an inflammatory response model was established based on IL-1 α stimulation of human gingival fibroblasts, and the same batch experiment as in example 1 was performed to study the anti-inflammatory effect of the composition, specifically as follows:
1. experimental Material
The cells used in the present invention are human gingival fibroblasts, batch No. 210708, commercially available. An inflammatory response model is obtained by induction with IL-1 alpha as a proinflammatory factor, the induction method can refer to the method described in the CN111139219A patent, and the anti-inflammatory effect of the composition can be effectively judged by detecting the content of PGE 2.
2. Experimental reagent
The present invention relates to reagents such as high-glucose DMEM (Gibco), fetal bovine serum (Lanzhou bright-colored), PBS (doctor's serum), MTT (Sigma), DMSO (Sigma), dexamethasone (Sigma), PGE2 ELISA kit (Abcam).
3. Testing instrument
The invention relates to CO 2 Incubator (Thermo, 150I), clean bench (Suzhou Antai, SW-CJ-1F), microplate reader (BioTek, Epoch).
4. Test method
(1) According to 4X 10 4 Cell/well seeding Density cells were plated onto 24-well plates, incubators (37 ℃, 5% CO) 2 ) And incubated overnight.
(2) The solutions of each experimental group and the control group were prepared according to the protocol shown in the following table, and based on the MTT test result (as shown in fig. 2), it was considered that the sample NANA PRO, which was based on gingival fibroblasts, did not exhibit significant cytotoxicity in the concentration range of 12.5 mg/mL.
TABLE 3 configuration schemes for the experimental and control groups
The configuration method of NANA PRO is as follows:
experimental group 1 sialic acid, trehalose and arginine were mixed according to SA: trehalose: the arginine proportion is as follows: 10:84.3: 5.7; mixing at a certain proportion, and pulverizing.
The experimental group 2 is obtained by weighing arginine and trehalose separately according to the proportion of the experimental group 1, mixing uniformly and then crushing.
(3) Administration and stimulation
According to the configuration scheme shown in table 3, when the cell plating rate in the 24-well plate reaches 40% -50%, the administration amount per well is 1mL, and 3 multiple wells are arranged in each group. In an incubator (37 ℃ C., 5% CO) 2 ) The medium was continued for 24 h.
4) Collecting cell supernatant: after 24h of culture, the cell culture supernatant was collected in an EP tube and stored frozen at-80 ℃ in a refrigerator.
5) And (3) ELISA detection: detection was performed according to the instructions for the PGE2 ELISA kit.
5. Result statistics and analysis
The results are expressed as Mean ± SD using GraphPad Prism mapping. The comparison between groups was performed by statistical analysis of t-test. Statistical analysis was two-tailed. P <0.05 was considered to have significant differences, and P <0.01 was considered to have very significant differences.
6. The result of the detection
The present invention gives the results shown in the following table:
TABLE 4 summary of PGE2 assay results for the experimental and control groups
Remarking: when the statistical analysis is carried out by using the t-test method, compared with the BC group, the significance is represented by #, the P-value <0.05 is represented by #, and the P-value <0.01 is represented by # #; significance was expressed as P-value <0.05 and P-value <0.01 compared to the NC group.
The results show that:
compared with the BC group, the secretion level of PGE2 in the NC group is extremely obviously increased (P <0.01), which indicates that the IL-1 alpha stimulation can extremely obviously improve the secretion amount of PGE2, and indicates that the IL-1 alpha stimulation condition is effective in the test.
The PGE2 secretion level was very significantly decreased in the PC group compared to the NC group (P <0.01), indicating that the positive control was effective in this test.
Compared with the NC group, the sample NANA PRO-12.5mg/mL can obviously inhibit the secretion of PGE2 caused by IL-1 alpha stimulation.
According to the MTT detection results, the NANA sample in the concentration range of 6.25mg/mL, the trehalose and arginine mixture in the concentration range of 12.5mg/mL and the NANA PRO in the concentration range of 12.5mg/mL are considered to be based on gingival fibroblasts, and no obvious cytotoxicity is shown.
The results of PGE2 tests in example 2 and example 1 were combined to show that sialic acid alone actually has a certain anti-inflammatory effect, but the present invention combines trehalose and arginine to obtain NANA PRO composition, in which the amount of sialic acid is reduced by one fifth, but the composition shows a more significant anti-inflammatory effect than the original effect of sialic acid and the blank group without sialic acid, indicating that the three components have a certain synergistic effect in anti-inflammatory.
Although the invention has been described in detail with respect to the general description and the specific embodiments thereof, it will be apparent to those skilled in the art that modifications and improvements can be made based on the invention. Accordingly, it is intended that all such modifications and alterations be included within the scope of this invention as defined in the appended claims.
Claims (10)
1. A sialic acid composition, comprising: sialic acid, trehalose, and arginine; the mass ratio of the sialic acid to the trehalose is 1: (8-10); the pH value of the water solution of the composition is 6-8.
2. The sialic acid composition of claim 1, wherein the sialic acid is present in the sialic acid composition at a weight percentage of at least 9%.
3. The sialic acid composition of claim 1, comprising 5 to 15 parts of sialic acid, 80 to 100 parts of trehalose, and 1 to 10 parts of arginine by weight.
4. The sialic acid composition of claim 3, comprising 9-11 parts of sialic acid, 80-90 parts of trehalose, and 5-6.5 parts of arginine by weight.
5. Use of the sialic acid composition of any one of claims 1 to 4 to alleviate inflammation.
6. Use of a sialic acid composition as claimed in any one of claims 1 to 4 in the manufacture of a medicament for reducing inflammation or a cosmetic product with soothing effect.
7. An anti-inflammatory product comprising a sialic acid composition as claimed in any one of claims 1 to 4.
8. An anti-inflammatory product according to claim 7, characterized in that it is a cosmetic product with soothing effect or a pharmaceutical preparation with anti-inflammatory function.
9. An anti-inflammatory product as in claim 7 or 8, further comprising: any one or more of stabilizers, solubilizers, buffers, preservatives, thickeners, abrasives, humectants, flavoring agents, buffers, or other excipients.
10. An anti-inflammatory product as claimed in claim 7 or 8, wherein the dosage form of the anti-inflammatory product is one or more of tablets, powders, granules, capsules, suppositories, aqueous solutions, ointments, creams, gels, inhalants, sprays, films, suspensions, lozenges, elixirs, syrups, water-in-oil or oil-in-water emulsions, oral liquids or drop pills.
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| AU2003200016A1 (en) * | 1998-07-27 | 2003-04-10 | E-L Management Corp. | Topical compositions containing sialyl sugars and their derivatives |
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| CN111139219A (en) * | 2020-01-09 | 2020-05-12 | 广东博溪生物科技有限公司 | Method and application of IL-1a for establishing human gingival fibroblast inflammation model |
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| CN113876665A (en) * | 2021-11-12 | 2022-01-04 | 湖北省麦诗特生物科技有限公司 | Face cream composition with anti-wrinkle effect and preparation method and application thereof |
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| AU2003200016A1 (en) * | 1998-07-27 | 2003-04-10 | E-L Management Corp. | Topical compositions containing sialyl sugars and their derivatives |
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