CN114939120B - 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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- CN114939120B CN114939120B CN202210518150.6A CN202210518150A CN114939120B CN 114939120 B CN114939120 B CN 114939120B CN 202210518150 A CN202210518150 A CN 202210518150A CN 114939120 B CN114939120 B CN 114939120B
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- trehalose
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- SQVRNKJHWKZAKO-UHFFFAOYSA-N beta-N-Acetyl-D-neuraminic acid Natural products CC(=O)NC1C(O)CC(O)(C(O)=O)OC1C(O)C(O)CO SQVRNKJHWKZAKO-UHFFFAOYSA-N 0.000 title claims abstract description 59
- 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 58
- 239000000203 mixture Substances 0.000 title claims abstract description 30
- 206010061218 Inflammation Diseases 0.000 title claims abstract description 10
- 230000004054 inflammatory process Effects 0.000 title claims abstract description 10
- 230000003110 anti-inflammatory effect Effects 0.000 claims abstract description 30
- 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
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products 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-WSWWMNSNSA-N 0.000 claims abstract description 21
- 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
- 239000004475 Arginine Substances 0.000 claims abstract description 18
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 claims abstract description 18
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 claims abstract description 10
- 239000007864 aqueous solution Substances 0.000 claims abstract description 5
- 239000002537 cosmetic Substances 0.000 claims description 8
- -1 abrasive Substances 0.000 claims description 6
- 239000000872 buffer Substances 0.000 claims description 4
- 230000000694 effects Effects 0.000 claims description 4
- 239000002775 capsule Substances 0.000 claims description 2
- 239000006071 cream Substances 0.000 claims description 2
- 239000003814 drug Substances 0.000 claims description 2
- 239000010408 film Substances 0.000 claims description 2
- 239000000796 flavoring agent Substances 0.000 claims description 2
- 235000013355 food flavoring agent Nutrition 0.000 claims description 2
- 239000000499 gel Substances 0.000 claims description 2
- 239000008187 granular material Substances 0.000 claims description 2
- 239000003906 humectant Substances 0.000 claims description 2
- 239000007788 liquid Substances 0.000 claims description 2
- 239000007937 lozenge Substances 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims description 2
- 239000002674 ointment Substances 0.000 claims description 2
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 2
- 239000000825 pharmaceutical preparation Substances 0.000 claims description 2
- 239000006187 pill Substances 0.000 claims description 2
- 239000000843 powder Substances 0.000 claims description 2
- 239000003755 preservative agent Substances 0.000 claims description 2
- 230000002335 preservative effect Effects 0.000 claims description 2
- 239000002904 solvent Substances 0.000 claims description 2
- 239000007921 spray Substances 0.000 claims description 2
- 239000003381 stabilizer Substances 0.000 claims description 2
- 239000000829 suppository Substances 0.000 claims description 2
- 239000000725 suspension Substances 0.000 claims description 2
- 239000006188 syrup Substances 0.000 claims description 2
- 235000020357 syrup Nutrition 0.000 claims description 2
- 239000003826 tablet Substances 0.000 claims description 2
- 239000002562 thickening agent Substances 0.000 claims description 2
- 239000002552 dosage form Substances 0.000 claims 1
- 239000000839 emulsion Substances 0.000 claims 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims 1
- 238000011160 research Methods 0.000 abstract description 3
- 238000013329 compounding Methods 0.000 abstract description 2
- 208000003265 stomatitis Diseases 0.000 abstract 1
- XEYBRNLFEZDVAW-ARSRFYASSA-N dinoprostone Chemical compound CCCCC[C@H](O)\C=C\[C@H]1[C@H](O)CC(=O)[C@@H]1C\C=C/CCCC(O)=O XEYBRNLFEZDVAW-ARSRFYASSA-N 0.000 description 16
- 229960002986 dinoprostone Drugs 0.000 description 16
- XEYBRNLFEZDVAW-UHFFFAOYSA-N prostaglandin E2 Natural products CCCCCC(O)C=CC1C(O)CC(=O)C1CC=CCCCC(O)=O XEYBRNLFEZDVAW-UHFFFAOYSA-N 0.000 description 16
- 230000000638 stimulation Effects 0.000 description 12
- 210000004027 cell Anatomy 0.000 description 10
- 238000000034 method Methods 0.000 description 9
- 238000001514 detection method Methods 0.000 description 8
- 210000002950 fibroblast Anatomy 0.000 description 8
- 230000028327 secretion Effects 0.000 description 8
- SQVRNKJHWKZAKO-PFQGKNLYSA-N N-acetyl-beta-neuraminic acid Chemical compound CC(=O)N[C@@H]1[C@@H](O)C[C@@](O)(C(O)=O)O[C@H]1[C@H](O)[C@H](O)CO SQVRNKJHWKZAKO-PFQGKNLYSA-N 0.000 description 7
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 6
- 102000004125 Interleukin-1alpha Human genes 0.000 description 6
- 108010082786 Interleukin-1alpha Proteins 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- 238000007619 statistical method Methods 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- 239000003153 chemical reaction reagent Substances 0.000 description 5
- 238000008157 ELISA kit Methods 0.000 description 4
- 238000000692 Student's t-test Methods 0.000 description 4
- 230000028709 inflammatory response Effects 0.000 description 4
- 238000012353 t test Methods 0.000 description 4
- 231100000002 MTT assay Toxicity 0.000 description 3
- 238000000134 MTT assay Methods 0.000 description 3
- 239000002253 acid Substances 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- 231100000135 cytotoxicity Toxicity 0.000 description 3
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- 238000002474 experimental method Methods 0.000 description 3
- 230000006870 function Effects 0.000 description 3
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- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- 238000002965 ELISA Methods 0.000 description 2
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- 238000004113 cell culture Methods 0.000 description 2
- 239000012228 culture supernatant Substances 0.000 description 2
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- UREBDLICKHMUKA-CXSFZGCWSA-N dexamethasone Chemical compound C1CC2=CC(=O)C=C[C@]2(C)[C@]2(F)[C@@H]1[C@@H]1C[C@@H](C)[C@@](C(=O)CO)(O)[C@@]1(C)C[C@@H]2O UREBDLICKHMUKA-CXSFZGCWSA-N 0.000 description 2
- 229960003957 dexamethasone Drugs 0.000 description 2
- 239000012091 fetal bovine serum Substances 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 239000008194 pharmaceutical composition Substances 0.000 description 2
- 238000007747 plating Methods 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- 230000000770 proinflammatory effect Effects 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 229940034610 toothpaste Drugs 0.000 description 2
- 239000000606 toothpaste Substances 0.000 description 2
- HVCOBJNICQPDBP-UHFFFAOYSA-N 3-[3-[3,5-dihydroxy-6-methyl-4-(3,4,5-trihydroxy-6-methyloxan-2-yl)oxyoxan-2-yl]oxydecanoyloxy]decanoic acid;hydrate Chemical class O.OC1C(OC(CC(=O)OC(CCCCCCC)CC(O)=O)CCCCCCC)OC(C)C(O)C1OC1C(O)C(O)C(O)C(C)O1 HVCOBJNICQPDBP-UHFFFAOYSA-N 0.000 description 1
- 229930186217 Glycolipid Chemical class 0.000 description 1
- 108090000288 Glycoproteins Chemical class 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 230000003925 brain function Effects 0.000 description 1
- 210000000133 brain stem Anatomy 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 150000002270 gangliosides Chemical class 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 210000004969 inflammatory cell Anatomy 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000002324 mouth wash Substances 0.000 description 1
- 229940051866 mouthwash Drugs 0.000 description 1
- 210000000653 nervous system Anatomy 0.000 description 1
- 108010008217 nidogen Proteins 0.000 description 1
- 239000007764 o/w emulsion Substances 0.000 description 1
- 229920001542 oligosaccharide Polymers 0.000 description 1
- 150000002482 oligosaccharides Chemical class 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000010298 pulverizing process Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 125000005629 sialic acid group Chemical group 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 230000032258 transport Effects 0.000 description 1
- 239000007762 w/o emulsion Substances 0.000 description 1
Classifications
-
- 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
-
- 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]
-
- 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
-
- 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)
-
- 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
-
- 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]
-
- 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
-
- 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
-
- 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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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
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- Pharmacology & Pharmacy (AREA)
- Medicinal Chemistry (AREA)
- Chemical & Material Sciences (AREA)
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- Birds (AREA)
- Pain & Pain Management (AREA)
- Rheumatology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Oral & Maxillofacial Surgery (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
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 sialic acid to trehalose is 1: (8-10); the pH value of the aqueous solution of the composition is 6-8. According to the research, sialic acid has a certain anti-inflammatory effect, and further, the composition compounded by sialic acid, trehalose and arginine is provided on the basis of the sialic acid, the trehalose and the arginine, and the anti-inflammatory effect is remarkably improved by compounding the sialic acid, the trehalose and the arginine, so that the sialic acid has an important significance in the anti-inflammatory application field, particularly in the field of reducing 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), a derivative of 9-carbon monosaccharides, also known as nidus Collocaliae, is a naturally occurring carbohydrate. Sialic acid is considered a transport transmitter of gangliosides and is one of the components of the brain, usually in the form of an oligosaccharide, glycolipid or glycoprotein.
The existing studies indicate that sialic acid plays a very important role in the production and development of brain and nervous system, and is important for normal development of brain function in children. In addition, sialic acid is widely used in various fields of life, for example, in various foods, cosmetics and health products.
However, studies on sialic acid function are currently inadequate, and their use in a broader range remains to be studied.
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, and 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 sialic acid to trehalose is 1: (8-10); the pH value of the aqueous solution of the composition is 6-8.
The invention discovers that sialic acid has a certain anti-inflammatory effect, and on the basis of the research, after the sialic acid, the trehalose and the arginine are compounded in the process of exploring more functions, the anti-inflammatory capability of the sialic acid is obviously improved. The specific combination mode is that the dosage of sialic acid and 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.
Further, in the sialic acid composition, the mass percent of sialic acid is more than or equal to 9%.
Further, the pharmaceutical composition comprises, by weight, 5-15 parts of sialic acid, 80-100 parts of trehalose and 1-10 parts of arginine.
Further, the pharmaceutical composition comprises 9-11 parts of sialic acid, 80-90 parts of trehalose and 5-6.5 parts of arginine by weight.
The invention further provides the use of the sialic acid composition in the alleviation of inflammation.
The invention further provides application of the sialic acid composition in preparing medicines for relieving inflammation or cosmetics with relieving effects.
In a second aspect, the invention provides an anti-inflammatory product comprising the sialic acid composition.
Further, the anti-inflammatory product is a cosmetic with soothing efficacy or a pharmaceutical preparation with anti-inflammatory function. As a specific cosmetic category, the cosmetic having soothing effect includes toothpaste having anti-inflammatory function.
Further, the method further comprises the following steps: any one or more of a stabilizer, solubilizer, buffer, preservative, thickener, abrasive, humectant, flavoring agent, buffer or other excipient.
Further, the anti-inflammatory product is in the form of one or more of a tablet, powder, granule, capsule, suppository, aqueous solution, ointment, cream, gel, inhalant, spray, film, suspension, lozenge, elixir, syrup, water-in-oil or oil-in-water emulsion, oral liquid or drop pill.
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 obtained sialic acid composition is obviously improved in the anti-inflammatory effect, and compared with an untreated inflammatory cell model, the reduction of the expression level of PGE2 is more than 35%, so that the sialic acid composition has important significance in the field of reducing inflammation. In particular, the invention establishes an inflammatory reaction model on human gingival fibroblasts, and proves that the composition has remarkable anti-inflammatory effect on the model, which proves that the composition can be more particularly applied to cosmetics or sanitary products such as toothpaste, mouthwash and the like.
Drawings
FIG. 1 is a schematic illustration of MTT assay results for sialic acid provided in example 1 of the present invention.
FIG. 2 is a schematic diagram showing the MTT test results of NANANANAPRO provided in example 2 of the present invention.
Detailed Description
The following examples are illustrative of the invention and are not intended to limit the scope of the invention.
The reagents used in the examples described below are commercially available unless otherwise specified.
The test methods described in the examples below can be carried out according to conventional methods in the art unless otherwise specified.
Example 1
The present example establishes an inflammatory response model based on IL-1 alpha stimulation of human gingival fibroblasts, and studies the anti-inflammatory effects of sialic acid, specifically as follows:
1. experimental materials
The cells used in the invention are human gingival fibroblasts, batch number 210708, and are commercially available. The IL-1 alpha is used as a pro-inflammatory factor to induce to obtain an inflammatory response model, the induction method can be referred to the method described in 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 sugar DMEM (Gibco), fetal bovine serum (bright red in langerhans), PBS (bosch), MTT (Sigma), DMSO (Sigma), dexamethasone (Sigma), and PGE2 ELISA kit (Abcam).
3. Test instrument
The present invention relates to CO 2 Incubator (Thermo, 150I), ultra clean bench (Suzhou Antai, SW-CJ-1F), microplate reader (BioTek, epoch).
4. Test method
(1) According to 4X 10 4 Seed density of cells/well cells were seeded into 24-well plates, incubator (37 ℃, 5% co) 2 ) Incubate overnight.
(2) 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 (nidulanic acid, nidogen, sialic acid) showed no significant cytotoxicity in the 6.25mg/mL concentration range based on gingival fibroblasts.
Table 1 configuration scheme for experimental and control groups
(3) Administration and stimulation
According to the configuration scheme of Table 1, when the cell plating rate in the 24-well plate reaches 40% -50%, group administration and stimulation are carried out, the administration amount of each well is 1mL, and 3 compound wells are arranged in each group. Then in incubator (37 ℃, 5% CO) 2 ) The culture was continued for 24 hours.
(4) Collecting cell supernatant
After 24h of incubation, the cell culture supernatant was collected in an EP tube and stored in a freezer at-80 ℃.
(5) ELISA detection
Detection was performed according to the instructions of the PGE2 ELISA kit.
5. Result statistics and analysis
GraphPad Prism was used to map and the results were expressed as mean±sd. Comparisons between groups were performed using t-test statistical analysis. Statistical analysis was double tailed. P <0.05 was considered to have significant differences and P <0.01 was considered to have very significant differences.
6. Detection result
The present invention gives the results shown in the following table:
TABLE 2 summary of PGE2 detection results for the experimental group and the respective control groups
Remarks: when the statistical analysis is performed by the t-test method, compared with the BC group, the significance is expressed as # and the P-value <0.05 is expressed as # and the P-value <0.01 is expressed as #; compared with NC groups, significance is expressed as x, P-value <0.05 is expressed as x, and P-value <0.01 is expressed as x.
The results showed that PGE2 secretion levels were significantly increased in NC group compared to BC group (P < 0.01), indicating that IL-1α stimulation could significantly increase PGE2 secretion, indicating that the present test IL-1α stimulation conditions were effective.
PGE2 secretion levels were significantly reduced in the PC group compared to NC group (P < 0.01), indicating that the positive control of this test was effective.
Compared with NC group, sample nidus Collocaliae acid (nidus Collocaliae acid, nidus Collocaliae extract, sialic acid) -6.25mg/mL can significantly inhibit PGE2 secretion caused by IL-1α stimulation.
Example 2
The present example establishes an inflammatory response model based on IL-1α stimulation of human gingival fibroblasts, and the same batch experiment as in example 1 is performed to study the anti-inflammatory effect of the composition, specifically as follows:
1. experimental materials
The cells used in the invention are human gingival fibroblasts, batch number 210708, and are commercially available. The IL-1 alpha is used as a pro-inflammatory factor to induce to obtain an inflammatory response model, the induction method can be referred to the method described in 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 sugar DMEM (Gibco), fetal bovine serum (bright red in langerhans), PBS (bosch), MTT (Sigma), DMSO (Sigma), dexamethasone (Sigma), and PGE2 ELISA kit (Abcam).
3. Test instrument
The present invention relates to CO 2 Incubator (Thermo, 150I), ultra clean bench (Suzhou Antai, SW-CJ-1F), microplate reader (BioTek, epoch).
4. Test method
(1) According to 4X 10 4 Seed density of cells/well cells were seeded into 24-well plates, incubator (37 ℃, 5% co) 2 ) Incubate overnight.
(2) Solutions of each of the experimental and control groups were prepared according to the protocol shown in the following table, and based on the MTT assay results (shown in fig. 2), it was considered that the sample NANA PRO showed no significant cytotoxicity in the 12.5mg/mL concentration range based on gingival fibroblasts.
Table 3 configuration scheme for experimental and control groups
The configuration method of NANA PRO is as follows:
experiment group 1 sialic acid, trehalose and arginine were prepared as per SA: trehalose: the arginine ratio is: 10:84.3:5.7; mixing the materials according to a certain proportion, and pulverizing.
And in the experimental group 2, arginine and trehalose are weighed according to the proportion of the experimental group 1, uniformly mixed and crushed to obtain the compound.
(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%, group administration and stimulation are carried out, the administration amount of each well is 1mL, and 3 compound wells are arranged in each group. In incubator (37 ℃, 5% CO) 2 ) The culture was continued for 24 hours.
4) Cell supernatant was collected: after 24h of incubation, the cell culture supernatant was collected in an EP tube and stored in a freezer at-80 ℃.
5) ELISA detection: detection was performed according to the instructions of the PGE2 ELISA kit.
5. Result statistics and analysis
GraphPad Prism was used to map and the results were expressed as mean±sd. Comparisons between groups were performed using t-test statistical analysis. Statistical analysis was double tailed. P <0.05 was considered to have significant differences and P <0.01 was considered to have very significant differences.
6. Detection result
The present invention gives the results shown in the following table:
TABLE 4 summary of PGE2 detection results for the experimental and control groups
Remarks: when the statistical analysis is performed by the t-test method, compared with the BC group, the significance is expressed as # and the P-value <0.05 is expressed as # and the P-value <0.01 is expressed as #; compared with NC groups, significance is expressed as x, P-value <0.05 is expressed as x, and P-value <0.01 is expressed as x.
The results show that:
compared with the BC group, the PGE2 secretion level of the NC group is extremely obviously increased (P < 0.01), which shows that the IL-1 alpha stimulation can extremely obviously increase the secretion amount of PGE2, and the condition of the IL-1 alpha stimulation tested in this time is effective.
PGE2 secretion levels were significantly reduced in the PC group compared to NC group (P < 0.01), indicating that the positive control of this test was effective.
Compared with NC group, the NANA PRO-12.5mg/mL can significantly inhibit PGE2 secretion caused by IL-1 alpha stimulation.
According to the MTT assay results, none of the samples NANA in the 6.25mg/mL concentration range, the trehalose and arginine mixture in the 12.5mg/mL concentration range, and NANA PRO in the 12.5mg/mL concentration range were considered to exhibit significant cytotoxicity based on gingival fibroblasts.
As can be seen from the results of PGE2 in comparative examples 2 and 1, sialic acid alone had a certain anti-inflammatory effect, but the composition NANA PRO of the present invention, on the basis of which trehalose and arginine were compounded, was reduced to one fifth in the amount of sialic acid component, but showed a more remarkable anti-inflammatory effect than the original effect of sialic acid and the blank group in which sialic acid component was omitted, indicating that the three had a certain synergistic effect in anti-inflammatory aspect.
While the invention has been described in detail in the foregoing general description and with reference to specific embodiments thereof, it will be apparent to one skilled in the art that modifications and improvements can be made thereto. Accordingly, such modifications or improvements may be made without departing from the spirit of the invention and are intended to be within the scope of the invention as claimed.
Claims (7)
1. A sialic acid composition with anti-inflammatory function, which is characterized by comprising sialic acid, trehalose and arginine; the mass ratio of sialic acid to trehalose is 1: (8-10); the pH value of the aqueous solution of the composition is 6-8;
9-11 parts of sialic acid, 80-90 parts of trehalose and 5-6.5 parts of arginine.
2. The sialic acid composition of claim 1, wherein the sialic acid is present in the sialic acid composition at a level of greater than or equal to 9% by mass.
3. Use of the sialic acid composition of claim 1 or 2 in the manufacture of a medicament for alleviating inflammation or a cosmetic having soothing efficacy.
4. An anti-inflammatory product comprising the sialic acid composition of claim 1 or 2.
5. The anti-inflammatory product according to claim 4, wherein the anti-inflammatory product is a cosmetic having soothing effect or a pharmaceutical preparation having anti-inflammatory function.
6. The anti-inflammatory product according to claim 4 or 5, further comprising: any one or more of a stabilizer, solubilizer, buffer, preservative, thickener, abrasive, humectant, flavoring agent, buffer or other excipient.
7. The anti-inflammatory product according to claim 4 or 5, wherein the anti-inflammatory product is in a dosage form selected from the group consisting 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 |
| CN113768808A (en) * | 2021-09-13 | 2021-12-10 | 武汉中科光谷绿色生物技术有限公司 | Sialic acid-containing composition and preparation method and application thereof |
| 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 |
| CN104586862A (en) * | 2015-02-06 | 2015-05-06 | 武汉中科光谷绿色生物技术有限公司 | Application of N-acetylneuraminic acid monomer, N-acetylneuraminic acid hydrate or N-acetylneuraminate in external anti-inflammation medicines |
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