CN117304056A - Silybum oil ceramide and its synthesis method and application - Google Patents

Silybum oil ceramide and its synthesis method and application Download PDF

Info

Publication number
CN117304056A
CN117304056A CN202310051508.3A CN202310051508A CN117304056A CN 117304056 A CN117304056 A CN 117304056A CN 202310051508 A CN202310051508 A CN 202310051508A CN 117304056 A CN117304056 A CN 117304056A
Authority
CN
China
Prior art keywords
ceramide
acid
silybum marianum
oil
marianum oil
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN202310051508.3A
Other languages
Chinese (zh)
Inventor
杨超文
叶柳
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Shenzhen Dikeman Biotechnology Co ltd
Original Assignee
Shenzhen Dikeman Biotechnology Co ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Shenzhen Dikeman Biotechnology Co ltd filed Critical Shenzhen Dikeman Biotechnology Co ltd
Priority to CN202310051508.3A priority Critical patent/CN117304056A/en
Priority to PCT/CN2023/133524 priority patent/WO2024109867A1/en
Publication of CN117304056A publication Critical patent/CN117304056A/en
Pending legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C233/00—Carboxylic acid amides
    • C07C233/01—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms
    • C07C233/16—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by singly-bound oxygen atoms
    • C07C233/17—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by singly-bound oxygen atoms with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by an acyclic carbon atom
    • C07C233/20—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by singly-bound oxygen atoms with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by an acyclic carbon atom having the carbon atom of the carboxamide group bound to a carbon atom of an acyclic unsaturated carbon skeleton
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/115—Fatty acids or derivatives thereof; Fats or oils
    • A23L33/12—Fatty acids or derivatives thereof
    • 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
    • A61K31/16—Amides, e.g. hydroxamic acids
    • A61K31/164—Amides, e.g. hydroxamic acids of a carboxylic acid with an aminoalcohol, e.g. ceramides
    • 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
    • A61K8/68—Sphingolipids, e.g. ceramides, cerebrosides, gangliosides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
    • 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
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P39/00—General protective or antinoxious agents
    • A61P39/06—Free radical scavengers or antioxidants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/08—Anti-ageing preparations
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C231/00—Preparation of carboxylic acid amides
    • C07C231/02—Preparation of carboxylic acid amides from carboxylic acids or from esters, anhydrides, or halides thereof by reaction with ammonia or amines
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C233/00—Carboxylic acid amides
    • C07C233/01—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms
    • C07C233/16—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by singly-bound oxygen atoms
    • C07C233/17—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by singly-bound oxygen atoms with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by an acyclic carbon atom
    • C07C233/18—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by singly-bound oxygen atoms with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by an acyclic carbon atom having the carbon atom of the carboxamide group bound to a hydrogen atom or to a carbon atom of an acyclic saturated carbon skeleton
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Medicinal Chemistry (AREA)
  • Dermatology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Epidemiology (AREA)
  • Mycology (AREA)
  • Polymers & Plastics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Food Science & Technology (AREA)
  • Nutrition Science (AREA)
  • Biophysics (AREA)
  • Toxicology (AREA)
  • Pain & Pain Management (AREA)
  • Rheumatology (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Biochemistry (AREA)
  • Gerontology & Geriatric Medicine (AREA)
  • Molecular Biology (AREA)
  • Birds (AREA)
  • Botany (AREA)
  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)

Abstract

The invention belongs to the technical field of biological medicines, and discloses silybum marianum oil ceramide which is obtained by reacting silybum marianum oil fatty acid with a sphingosine compound, wherein the sphingosine compound is selected from sphingosine, phytosphingosine and dihydrosphingosine. The silybum marianum oil ceramide has excellent performances in the aspects of repairing natural skin barriers, resisting inflammation, healing tissues, resisting aging and the like, and has wide application prospects in the fields of cosmetics, health-care products, biological medicines and the like.

Description

Silybum oil ceramide and its synthesis method and application
Technical Field
The invention belongs to the technical field of biological medicines, and particularly relates to silybum marianum oil ceramide and a synthesis method and application thereof.
Background
Ceramides (ceramides, also known as molecular nails) naturally occur in the skin and are very important components of the skin barrier (stratum corneum), in amounts of up to 40-50 wt.%, ceramides are a class of sphingolipids consisting of long-chain bases of sphingosine and fatty acids, in which the carbon chain length, unsaturation and number of hydroxyl groups of the sphingosine moiety, fatty acid moiety are all variable, and ceramides represent a class of compounds. Ceramide has excellent properties in regulating skin barrier function, recovering skin moisture, enhancing adhesion between skin keratinocytes, and the like.
Because of the importance of ceramides, many cosmetic and pharmaceutical companies are researching and developing corresponding products. The natural plant-derived ceramide can form an effective skin barrier to prevent water loss and resist external damage due to the more sustainable and more environment-friendly raw material source and the characteristics similar to the skin ceramide components, and can become a next-generation environment-friendly, safe and reliable ceramide product.
The silybum marianum oil is an excellent medicinal vegetable oil extracted from silybum marianum. The silybum marianum oil is mainly linoleic acid and contains rich fatty acids such as oleic acid, palmitic acid, stearic acid, arachidic acid, behenic acid, linolenic acid and the like. In addition to fatty acids, silybum marianum oil contains various hydrocarbons, triacontanol, beta-sitosterol, campesterol, etc. The silybum marianum oil has light color, low viscosity and high nutritive value, has good effects of resisting oxidation, inflammation and aging, and is a good material for manufacturing advanced cosmetics. In industry, the silybum marianum oil is better drying oil, has excellent color retention and durability, and is a raw material for manufacturing paint varnish; in traditional medicine, silybum marianum oil is also used for preventing and treating diseases such as atherosclerosis, hypertension, coronary heart disease, light and medium tuberculosis and the like.
Disclosure of Invention
The invention aims to provide ceramide synthesized by using silybum marianum oil fatty acid of plant origin.
Another object of the present invention is to provide a method for synthesizing silybum marianum oil ceramide, which uses silybum marianum oil fat or silybum marianum oil fatty acid which is natural plant source and is easily obtained as raw material.
It is a further object of the present invention to provide the use of silybum marianum oil ceramide.
In order to achieve one of the above purposes, the present invention adopts the following technical scheme:
in a first aspect of the invention, silybum marianum oil ceramide is obtained by reacting silybum marianum oil fatty acid with a sphingosine compound selected from the group consisting of sphingosine, phytosphingosine and sphinganine.
The reaction can be chemical synthesis reaction (as detailed below), or microbial fermentation method, i.e. using Pichia pastoris or Saccharomyces cerevisiae, fermenting under certain environment to obtain sphingosine compound, and adding fatty acid to obtain ceramide; or using silybum marianum oil as raw material, selecting proper strain, and making fermentation so as to obtain silybum marianum oil ceramide.
Sphingosine refers to 2-amino-4-octadecene-1, 3-diol, phytosphingosine refers to 2-amino-octadecane-1, 3, 4-triol, and dihydrosphingosine refers to 2-amino-octadecane-1, 3-diol.
Further, the silybum marianum oil fatty acid is obtained by hydrolyzing silybum marianum oil fat.
Further, the silybum marianum oil fatty acid contains 50-75wt% of linoleic acid.
Further, the silybum marianum oil fatty acid contains 15-45 wt% of oleic acid.
Further, the silybum marianum oil fatty acid contains 1-10wt% of palmitic acid.
Further, the silybum marianum oil fatty acid contains 1-6wt% of stearic acid.
Further, the silybum marianum oil fatty acid contains 0.2-4wt% of arachidic acid.
Further, the silybum marianum oil fatty acid contains 0.5-4wt% of behenic acid.
Further, the silybum marianum oil fatty acid contains 0.01-2.5 wt% of linolenic acid.
In addition, the silybum marianum oil fatty acid also contains 0-1.5 wt% of myristic acid.
The composition of the silybum marianum oil fatty acid is as follows: 50 to 75 weight percent of linoleic acid, 15 to 45 weight percent of oleic acid, 1 to 10 weight percent of palmitic acid, 1 to 6 weight percent of stearic acid, 0.2 to 4 weight percent of arachidic acid, 0.5 to 4 weight percent of behenic acid, 0.01 to 2.5 weight percent of linolenic acid and 0 to 1.5 weight percent of myristic acid.
The main component of the silybum marianum oil fatty acid is linoleic acid, other fatty acids comprise oleic acid, palmitic acid, stearic acid, arachidic acid, behenic acid and linolenic acid, which are essential components, are influenced by plant varieties, soil, climate, production places, picking seasons and extraction processes, the content of each component is different, and myristic acid is not necessarily contained, and is an optional component or an unnecessary component.
Silybum marianum oil ceramide, which comprises the following components: linoleic acid ceramide, oleic acid ceramide, palmitic acid ceramide, stearic acid ceramide, arachidic acid ceramide, behenic acid ceramide, linolenic acid ceramide; since fatty acids all participate in the same reaction, the mass ratio of ceramide after the reaction is not greatly changed, so the composition of silybum marianum oil ceramide is similar to that of silybum marianum oil fatty acid: 50 to 75 weight percent of linoleic acid ceramide, 15 to 45 weight percent of oleic acid ceramide, 1 to 10 weight percent of palmitic acid ceramide, 1 to 6 weight percent of stearic acid ceramide, 0.2 to 4 weight percent of arachidic acid ceramide, 0.5 to 4 weight percent of behenic acid ceramide and 0.01 to 2.5 weight percent of linolenic acid ceramide. The content of each component is different due to the different content of each fatty acid in the silybum marianum oil fatty acid or oil. In addition, the silybum marianum oil ceramide also comprises ceramide obtained by reacting myristic acid with sphingosine compounds, namely 0-1.5 wt% of myristic acid ceramide. Silybum oil ceramide also includes hydrocarbons, triacontanol, beta-sitosterol, campesterol, etc. compounds that are present in the fatty acids of Silybum oil but do not react with sphingosine compounds.
Silybum marianum oil ceramide, which comprises the following components: linoleic acid ceramide, oleic acid ceramide, palmitic acid ceramide, stearic acid ceramide; 50-75wt% of linoleic acid ceramide, 15-45wt% of oleic acid ceramide, 1-10wt% of palmitic acid ceramide and 1-6wt% of stearic acid ceramide.
Further, the silybum marianum oil ceramide comprises arachidic acid ceramide, and the arachidic acid ceramide accounts for 0.2-4wt%.
Further, the silybum marianum oil ceramide comprises behenic acid ceramide, and the behenic acid ceramide accounts for 0.5-4wt%.
Further, the silybum marianum oil ceramide comprises linolenic acid ceramide with the proportion of 0.01-2.5 wt%.
Further, the silybum marianum oil ceramide comprises not more than 1.5wt% of myristic acid ceramide, especially 0.1 to 1.5wt% of myristic acid ceramide.
The linoleic acid ceramide is obtained by condensation reaction of linoleic acid and sphingosine compounds, and comprises linoleic acid phytosphingosine ceramide, linoleic acid sphingosine ceramide and linoleic acid dihydrosphingosine ceramide; oleic acid ceramide is obtained by condensation reaction of oleic acid and sphingosine compounds, and comprises oleic acid phytosphingosine ceramide, oleic acid sphingosine ceramide and oleic acid dihydrosphingosine ceramide; the palmitic acid ceramide is obtained by condensation reaction of palmitic acid and sphingosine compounds, and comprises palmitic acid phytosphingosine ceramide, palmitic acid sphingosine ceramide and palmitic acid dihydrosphingosine ceramide; stearic acid ceramide is obtained by condensation reaction of stearic acid and sphingosine compound, and comprises stearic acid phytosphingosine ceramide, stearic acid sphingosine ceramide and stearic acid dihydrosphingosine ceramide; eicosanoic acid ceramide, behenic acid ceramide, linolenic acid ceramide, myristic acid ceramide, and the like.
In a second aspect of the invention, a method for synthesizing silybum marianum oil ceramide comprises the following steps:
under the conditions of condensing agent and organic alkali, the silybum marianum oil fatty acid reacts with the sphingosine compound, the condensing agent is EDCI, and the organic alkali is DIPEA.
Further, the molar ratio of the silybum marianum oil fatty acid to the sphingosine compound to the EDCI to the DIPEA is 1: (1-1.5): (1-2): (1-2), wherein the solvent for the reaction is at least one of dichloromethane, tetrahydrofuran, ethyl acetate and acetonitrile.
The commercially available silybum marianum oil is generally in the form of oil and needs to be hydrolyzed into silybum marianum oil fatty acid, and therefore the method further comprises the following steps:
the silybum marianum oil fat is hydrolyzed by saponification reaction to obtain silybum marianum oil fatty acid.
Further, the saponification reaction is hydrolysis of silybum marianum oil fat in potassium hydroxide solution.
Further, the mass ratio of the silybum marianum oil grease to the potassium hydroxide is 1: (1-2).
In a third aspect of the invention, the use of silybum marianum oil ceramide in cosmetics, medicines, dietary or health care products.
Further, the silybum marianum oil ceramide has at least one of skin barrier repair, tissue healing, anti-aging, anti-inflammatory, anti-photoaging, antioxidant, collagen synthesis promoting, elastin activity maintaining and whitening effects.
A composition comprising silybum marianum oil ceramide, said composition having at least one of skin barrier repair, tissue healing, anti-aging, anti-inflammatory, anti-photoaging, anti-oxidant, collagen synthesis promoting, elastin activity maintenance, whitening efficacy.
The composition contains acceptable auxiliary materials, including one or more of solubilizer, antiseptic, antioxidant, pH regulator, penetration enhancer, liposome, humectant, thickener, chelating agent, skin feel regulator, surfactant, emulsifier, essence and pigment; the composition is in the form of cream, emulsion, solution, film, aerosol or spray.
The invention has the following beneficial effects:
the silybum marianum oil fatty acid belongs to naturally-formed fatty acid, the main component is unsaturated fatty acid-linoleic acid, and in addition, oleic acid and saturated fatty acid such as palmitic acid and stearic acid are contained, and the silybum marianum oil ceramide is prepared by mild reaction with a sphingosine compound naturally existing in skin, has excellent performance in the aspects of repairing a natural skin barrier, resisting oxidation, resisting aging and the like, and has wide application prospects in the fields of cosmetics, health-care products, biological medicines and the like.
1. Better results compared to ceramide alone. Different ceramides have different effects due to the structural differences, and ceramides with a single structure generally have difficulty in having comprehensive effects. The scheme is based on a bionic thought, and the natural-source silybum marianum oil grease or fatty acid is used as a raw material to synthesize the compound ceramide so as to make up the difference of different ceramide effects, and trace fatty acid in the silybum marianum oil can form trace ceramide to play a role in efficacy supplement.
2. Compared with the compounded ceramide, the effect is better. Besides fatty acid (or grease), the silybum marianum oil is also rich in various hydrocarbons, triacontanol, beta-sitosterol, campesterol and the like, and the nutrient substances have good antioxidant, anti-inflammatory and anti-aging effects. The ceramide synthesized by the silybum marianum oil has a synergistic effect with other active ingredients contained in the silybum marianum oil, and has better effect compared with the ceramide compounded according to similar proportion.
3. The cost is lower. The method of the invention can rapidly obtain the composition compounded by various ceramides, and the vegetable-source silybum marianum oil or the fatty acid thereof has wide sources, is easy to obtain commercially, has lower cost, is more environment-friendly and economical, is different from the idea of mixing and compounding different single ceramides, has high raw material price, needs to separately produce different ceramides, and then is compounded, thereby increasing the preparation cost.
4. The synthesis method is simple. The method can adopt chemical synthesis to realize one-step preparation of various ceramides, and can also use a microbial fermentation method.
Drawings
FIGS. 1 and 2 are bar graphs showing the results of the cell proliferation activity test of example 4;
FIG. 3 is the result of the cell migration ability test of example 5;
FIG. 4 is a bar graph of elastase inhibition of example 6;
FIG. 5 is a bar graph showing the detection of IL-6 factor expression level in anti-inflammatory repair efficacy in example 7;
FIGS. 6 and 7 are bar charts of MMP1 expression levels in the photo-aging test of example 8;
FIG. 8 is a bar graph of DPPH radical scavenging for oxidation resistance test of example 9;
fig. 9 is a bar graph of the whitening activity test melanin content of example 10.
Detailed Description
The invention will be further illustrated with reference to specific examples.
EDCI refers to 1-ethyl- (3-dimethylaminopropyl) carbodiimide and DIPEA refers to N, N-diisopropylethylamine. The silica gel column chromatography uses Qingdao ocean silica gel (particle size 0.040-0.063 mm). Thin Layer Chromatography (TLC) using 60F254 silica gel plates was performed using UV light (254 nm) or iodine.
Example 1
Synthesis of ceramide from silybum marianum oil fatty acid and phytosphingosine
The first step: 50g of silybum marianum oil is dissolved in 60mL of tetrahydrofuran, cooled in an ice bath, 100mL of potassium hydroxide (25 wt%) solution is added dropwise, and the mixture is cooled to room temperature for reaction after the dropwise addition until TLC detection is completed.
Post-treatment: adding dilute hydrochloric acid (3N) to adjust the pH value of the reaction system to 3, adding 150mL of ethyl acetate to extract a water phase, adding 100mL of saturated saline water for washing once, and adding anhydrous Na into an organic phase 2 SO 4 Drying, filtering and concentrating in vacuum to obtain 40.8g of silybum marianum oil fatty acid.
And a second step of: silybum marianum oil fatty acid (50 mmol based on main component fatty acid), EDCI (65 mmol), DIPEA (65 mmol) were added to a 250mL round bottom flask, 100mL dichloromethane was added, followed by stirring at room temperature for 1 hour, and phytosphingosine (55 mmol) was added to the reaction system, followed by stirring at room temperature until TLC detection was complete.
Post-treatment: adding water for quenching reaction, separating an organic layer, drying, filtering and concentrating in vacuum, washing by a solvent to obtain silybum marianum oil ceramide, and analyzing a product by HPLC (high performance liquid chromatography) under the condition of HPLC chromatography: using an shimadzu high performance liquid chromatograph (LC-2030 c3d Plus), column temperature with Innoval ODS-2.6x250 mm,5 μm column: 30 ℃, sample injection volume: 10 μl, flow rate: 1.0mL/min, evaporation temperature: 40 ℃, carrier gas flow rate: 2.5L/min, mobile phase: 100% methanol.
The retention time of each component HPLC was: linolenic acid-phytosphingosine ceramide 8.2min, linoleic acid-phytosphingosine ceramide 9.5min, palmitic acid-phytosphingosine ceramide 10.7min, oleic acid-phytosphingosine ceramide 11.3min, stearic acid-phytosphingosine ceramide 13.9min, arachidic acid-phytosphingosine ceramide 16.5min, and behenic acid-phytosphingosine ceramide 18.3min.
The obtained products are analyzed by high performance liquid chromatography, the contents of linoleic acid-phytosphingosine ceramide, oleic acid-phytosphingosine ceramide, palmitic acid-phytosphingosine ceramide, stearic acid-phytosphingosine ceramide, arachidic acid-phytosphingosine ceramide and behenic acid-phytosphingosine ceramide are 65%, 18%, 5%, 3%, 2%, 1% and the balance of other components in sequence, and the contents are less.
Example 2
Synthesis of ceramide from silybum marianum oil fatty acid and sphingosine
The first step: 50g of silybum marianum oil is dissolved in 60mL of tetrahydrofuran, cooled in an ice bath, 100mL of potassium hydroxide (25 wt%) solution is added dropwise, and the mixture is cooled to room temperature for reaction after the dropwise addition until TLC detection is completed.
Post-treatment: adding dilute hydrochloric acid (3N) to adjust the pH value of the reaction system to 3, and adding 150The aqueous phase was extracted with mL of ethyl acetate, 100mL of saturated brine was added and washed once with water, and the organic phase was added to anhydrous Na 2 SO 4 Drying, filtering and concentrating in vacuum to obtain 41g of silybum marianum oil fatty acid.
And a second step of: silybum marianum oil fatty acid (50 mmol, based on main component fatty acid), EDCI (80 mmol), DIPEA (80 mmol) were added to a 250mL round bottom flask, 100mL dichloromethane was added, followed by stirring at room temperature for 1 hour, and then sphingosine (70 mmol) was added to the reaction system, followed by stirring at room temperature until TLC detection was complete.
Post-treatment: adding water for quenching reaction, separating an organic layer, drying, filtering and concentrating in vacuum, washing by a solvent to obtain silybum marianum oil ceramide, and analyzing a product by HPLC (high performance liquid chromatography) under the condition of HPLC chromatography: using an shimadzu high performance liquid chromatograph (LC-2030 c3d Plus), column temperature with Innoval ODS-2.6x250 mm,5 μm column: 30 ℃, sample injection volume: 10 μl, flow rate: 1.0mL/min, evaporation temperature: 40 ℃, carrier gas flow rate: 2.5L/min, mobile phase: 100% methanol.
The retention time of each component HPLC was: linolenic acid-sphingosine ceramide 7.9min, myristic acid-sphingosine ceramide 8.3min, linoleic acid-sphingosine ceramide 8.7min, oleic acid-sphingosine ceramide 10.2min, palmitic acid-sphingosine ceramide 10.4min, stearic acid-sphingosine ceramide 13.6min, arachidic acid-sphingosine ceramide 17.8min, behenic acid-sphingosine ceramide 24.1min.
The obtained products are analyzed by high performance liquid chromatography, and the contents of linoleic acid-sphingosine ceramide, oleic acid-sphingosine ceramide, palmitic acid-sphingosine ceramide, stearic acid-sphingosine ceramide, arachidic acid-sphingosine ceramide, behenic acid-sphingosine ceramide, linolenic acid-sphingosine ceramide and myristic acid-sphingosine ceramide are 52%, 20%, 9%, 6%, 4%, 2% and 1% in sequence, and the rest are other components, and the content is low.
Example 3
Synthesis of ceramide from Silybum oil fatty acid and sphinganine
The first step: 50g of silybum marianum oil is dissolved in 60mL of tetrahydrofuran, cooled in an ice bath, 100mL of potassium hydroxide (25 wt%) solution is added dropwise, and the mixture is cooled to room temperature for reaction after the dropwise addition until TLC detection is completed.
Post-treatment: adding dilute hydrochloric acid (3N) to adjust the pH value of the reaction system to 3, adding 150mL of ethyl acetate to extract a water phase, adding 100mL of saturated saline water for washing once, and adding anhydrous Na into an organic phase 2 SO 4 Drying, filtering and concentrating in vacuum to obtain 40.5g of silybum marianum oil fatty acid.
And a second step of: silybum marianum oil fatty acid (50 mmol, based on main component fatty acid), EDCI (70 mmol), DIPEA (70 mmol) were added to a 250mL round bottom flask, 100mL dichloromethane was added, followed by stirring at room temperature for 1 hour, then sphinganine (60 mmol) was added to the reaction system, and stirring at room temperature was performed until TLC detection was complete.
Post-treatment: adding water for quenching reaction, separating an organic layer, drying, filtering and concentrating in vacuum, washing by a solvent to obtain silybum marianum oil ceramide, and analyzing a product by HPLC (high performance liquid chromatography) under the condition of HPLC chromatography: using an shimadzu high performance liquid chromatograph (LC-2030 c3d Plus), column temperature with Innoval ODS-2.6x250 mm,5 μm column: 30 ℃, sample injection volume: 10 μl, flow rate: 1.0mL/min, evaporation temperature: 40 ℃, carrier gas flow rate: 2.5L/min, mobile phase: 100% methanol.
The retention time of each component HPLC was: linolenic acid-sphinganine ceramide 8.2min, myristic acid-sphinganine ceramide 8.5min, linoleic acid-sphinganine ceramide 9.5min, palmitic acid-sphinganine ceramide 10.6min, oleic acid-sphinganine ceramide 11.1min, stearic acid-sphinganine ceramide 13.5min, arachidic acid-sphinganine ceramide 16.0min, behenic acid-sphinganine ceramide 21.9min.
The obtained products are analyzed by high performance liquid chromatography, the contents of linoleic acid-dihydrosphingosine ceramide, oleic acid-dihydrosphingosine ceramide, palmitic acid-dihydrosphingosine ceramide, stearic acid-dihydrosphingosine ceramide, arachidic acid-dihydrosphingosine ceramide, behenic acid-dihydrosphingosine ceramide, linolenic acid-dihydrosphingosine ceramide and myristic acid-dihydrosphingosine ceramide are 56%, 29%, 6%, 2%, 1% and 1% in sequence, and the rest are other components, and the contents are less.
Example 4
MTT method for detecting proliferation activity of compound on cell
HaCaT cells were grown at 1X 10 4 The density of individuals/wells was seeded in 96-well plates and the incubator was overnight. After 24h, the supernatant was discarded, 100. Mu.L of medium containing samples of different concentrations (product of example 1) was added, incubation was continued for 24h, medium was removed, 100. Mu.L of thiazole blue (MTT) was added to each well, absorbance at 450nm was measured, and cell viability = A was calculated Drug delivery hole /A Blank hole ×100％。
As shown in figure 1, the silybum marianum oil ceramide has a promoting effect on cell viability, and the cell viability is 114.17%, 110.45%, 109.29%, 109.76%, 105.92%, 105.23%, 108.13%, 111.03% and 130.31% respectively at the concentrations of 3.90625, 7.8125, 15.625, 31.25, 62.5, 125, 250, 500 and 1000mg/L, the effective concentration is as low as 4mg/L, the safety concentration is 1000mg/L, the concentration gradient is relatively stable, the obvious effect of promoting cell proliferation is shown, and the tissue repair capability is good.
The proliferation activity of ceramide 2 on cells was measured in the same manner, and as a result, as shown in FIG. 2, cell viability was 61.49%, 60.03%, 55.41%, 54.64%, 53.37%, 46.95%, 44.05%, 40.35%, 39.42% at concentrations of 3.90625, 7.8125, 15.625, 31.25, 62.5, 125, 250, 500, 1000mg/L, respectively, which had an inhibitory effect on cell proliferation, and tissue repair potential was inferior to that of silybum marianum ceramide.
Example 5
Assessment of skin barrier repair by cell migration
Principle of: when the cells grow to be fused into a single-layer state, a scratch tool is manufactured on the fused single-layer cells, the cells in the blank area are removed by mechanical force, the migration condition of the cells to the cell-free area is observed through a period of culture, and the migration capability of the cells is reflected by measuring the migration distance of the cells.
The operation steps are as follows:
1. the culture plate is streaked. Firstly, a Marker pen is used for uniformly scribing transverse lines by comparing with a straight ruler, and the transverse lines are crossed through the through holes at intervals of about 0.5 cm to 1cm, and each hole at least passes through 5 lines, so that attention lines are not too thick when scribing.
2. And (5) paving cells. About 5X 10 is added to the well 5 Individual cells (the number of different cells is different, and the cell growth speed is regulated), and the inoculation principle is that the fusion rate reaches 100% after overnight.
3. Cell streaking. The next day the tip is used to scratch the cell layer along the line marked on the back of the plate on the first day, perpendicular to the cell plane (the same tip is preferably used between the different wells).
4. Washing cells. After the streaking was completed, cells were washed 3 times with sterile PBS, cells that did not adhere to the wall, i.e., streaked cells at streaking, and the gap left after streaking was clearly visible, followed by replacement of fresh serum-free medium.
5. And (5) culturing and observing the cells. After the sample (product of example 1, ceramide 3B) was diluted with the medium (product of example 1 at 5mg/L, ceramide 3B at 100 mg/L), the cells were placed in a cell culture dish at 37℃and 5wt% CO 2 Incubator culture, after 24 hours, cells were removed, observed with a microscope and the width of scratches was measured, and photographed, and the healing rate was calculated using Image J software.
The results are shown in fig. 3, and the scratch width of the experimental group is narrower than that of the solvent control group, which indicates that the silybum marianum oil ceramide has better tissue healing capacity. The healing rate of the solvent control group after 24 hours is 27.33%, the healing rate of silybum marianum oil ceramide after 24 hours is 90.23%, and the healing rate of ceramide 3B after 24 hours is 59.32%. The compound obviously improves the cell healing rate, has good skin tissue repair activity and has better effect than ceramide 3B.
Example 6
Elastase inhibition experiment tests anti-aging effect
Elastase inhibition method: 2mg/mL elastase solution (product of example 1) is taken, samples with different concentrations (2 mL) are added, vortex mixing is carried out fully, shaking is carried out for 20min at 37 ℃ by a 400r/min shaking table, 5mL of 0.5mol/L phosphate buffer solution with pH of 6.0 is added immediately, vortex mixing is carried out, a proper amount of mixed solution is taken into a 2mL centrifuge tube, centrifugation is carried out for 10min at 9 391×g, 200 mu L of supernatant is sucked into a 96-well plate precisely, absorbance is measured by an enzyme-labeled instrument at a wavelength of 495nm, and spectrum scanning at 400-800 nm is carried out simultaneously.
The substrate enzyme adding solution is used as a blank control group, the substrate enzyme adding and sample solution is used as an enzyme inhibition group, and the substrate enzyme adding and sample solution is used as a background. Each group is provided with 3 multiple holes. Inhibition ratio (%) = [1- (An-An ')/(A0-A0') ] ×100%, where A0 is absorbance with no enzyme added to the sample, A0 'is absorbance with no enzyme added to the substrate and no sample added to the enzyme, an is absorbance with only sample solution, an' is absorbance with no enzyme added to the sample. When An ' > An, the effect is expressed as acceleration, and the acceleration rate (%) = [1- (An ' -An)/(A0-A0 ') ] ×100%.
As shown in FIG. 4, silybum marianum oil ceramide has good inhibition effect on elastase at different concentrations, specifically, the inhibition rate of elastase at the concentration of 0.25g/L is 7.13%, the inhibition rate of elastase at the concentration of 0.5g/L is 13.60%, the inhibition rate of elastase at the concentration of 1.0g/L is 21.67%, and the inhibition rate of elastase at the concentration of 2.0g/L is 30.67%.
Example 7
LPS induced cell method for detecting anti-inflammatory repair efficacy
B16 mouse melanoma cells were grown at a density of 1X 10 4 The cells/wells were seeded in 96-well plates, placed in an incubator overnight, the supernatant was discarded after 24 hours, 100. Mu.L of samples of different concentrations diluted with DMEM medium (product of example 1) were added, the negative control group was DMEM medium without samples, 3 wells per group, and mixed with CO at 5 wt.% 2 Incubate at 37 ℃. Lipopolysaccharide model group and experimental group were added with 10 μg/mL LPS and incubated together for 24h 2h after dosing. After the reaction, 50. Mu.L of the cell supernatant was collected, and the intracellular IL-6 gene expression was detected using an IL-6ELISA kit.
The results are shown in FIG. 5, where IL-6 levels were 10.16 times the basal levels at a working concentration of 10. Mu.g/mL of LPS stimulation. Under the action of silymarin oil ceramide with the concentration of 50mg/L, 100mg/L, 200mg/L and 400mg/L respectively, the IL-6 factor level is obviously reduced and is 0.70 times, 0.45 times, 0.27 times and 0.09 times of that of an LPS model group respectively, and the silymarin oil ceramide is dose-dependent, so that the silymarin oil ceramide has good anti-inflammatory effect and can promote the repair of inflammatory damaged skin.
Example 8
MMP1 is also called interstitial collagenase and matrix metalloproteinase, belongs to matrix metalloproteinase family, and its main acting substrate is fibrous collagen, which can degrade collagen fiber and gelatin in extracellular matrix and change microenvironment of cells. MMP1 plays an important role in elastin, inhibiting MMP1 can improve the synthesis of fibroblast collagen and elastin, and reducing MMP activity can increase the collagen synthesis speed.
HaCaT cells were grown at 1X 10 5 The density of individuals/wells was seeded in 96-well plates and the incubator was overnight. After 24h, the supernatant was discarded, 100. Mu.L of medium containing samples of different concentrations (product of example 1) was added, no samples were added to the model group, the negative control group was DMEM medium without samples, 3 wells per group, and the mass fraction was 5% CO 2 After incubation for 2h at 37℃either UVA or UVB ultraviolet radiation is irradiated. The distance between the ultraviolet radiation source and the cells was 15cm, and the UVA intensity was 200mJ/cm 2 The irradiation time was 2 hours, and the UVB intensity was 50mJ/cm 2 The irradiation time was 1h. After the end of irradiation, incubation was continued for 12h in the incubator. Intracellular MMP-1 gene expression was detected using an MMP-1ELISA kit. Inhibition = 1- (experimental group MMP1 expression level/model group MMP1 expression level) ×100%.
As shown in fig. 6 and 7, the expression level of MMP1 in the negative control group was 1, the expression level in the model group was 1.90, and the inhibition ratio of MMP1 expression in the model group was 34%, 51%, 69% at the concentrations of 125, 250, 400mg/L of silymarin oil ceramide; for UVB, the MMP1 expression level of the negative control group was 1, the expression level of the model group was 2.33, and the inhibition rate of the silybum marianum oil ceramide expression was 44%, 52% and 65% relative to the model group at the concentrations of 125, 250 and 400 mg/L.
After UVA uv radiation, keratinocytes promote elevated expression of MMP1 by fibroblasts, thereby causing degradation of the extracellular matrix of the skin and collagen of the skin, leading to photoaging of the skin. The results show that the silybum marianum oil ceramide can inhibit the fibroblast from producing MMP1 caused by ultraviolet radiation, and has a certain effect on preventing skin photoaging.
Example 9
DPPH free radical scavenging detection of antioxidant performance
DPPH is 1, 1-diphenyl-2-trinitrophenylhydrazine, and can be used for antioxidant experiments.
Samples (product of example 1) at corresponding concentrations (50, 100, 200, 400, 800 mg/L) were mixed with 0.1mol/L DPPH, absolute ethanol solution at a ratio of 1:1, and DPPH and absolute ethyl alcohol 1:1, and the absorbance at 517 nm. The absorbance of the sample and the reaction solution of DPPH was designated as A1, the absorbance of the sample and the reaction solution of absolute ethyl alcohol was designated as A2, the absorbance of the reaction solution of DPPH and absolute ethyl alcohol was designated as A3, and the clearance rate of DPPH of the sample was = [1- (A1-A2)/A3 ]. Times.100%.
As a result, as shown in FIG. 8, the DPPH radical scavengers at concentrations of 50, 100, 200, 400 and 800mg/L were 10.00%, 11.45%, 13.77%, 19.77% and 23.55%, respectively, and excellent antioxidant effects were exhibited.
Example 10
Whitening Activity test
Taking B16 cells in exponential growth phase, digesting with trypsin-EDTA with mass fraction of 0.25%, blowing uniformly, and mixing the cells according to 3×10 5 Density of individual/well was seeded in 12-well plates. At 37 ℃, the mass fraction of CO is 5 percent 2 Incubated overnight in the environment. Removing supernatant, adding culture solution containing samples with different mass concentrations (product of example 1), incubating with RPMI-1640 culture medium without sample as blank group, incubating with DMEM culture medium as mould group, and incubating with 3 compound holes in each group at mass fraction of 5% CO 2 Incubation was carried out for 24h at 37 ℃. The culture medium in the well plate was discarded, and after washing with Phosphate Buffer (PBS) one to two times, 1mL of N containing 10% by mass of DMSO was addedThe aOH solution (1 mol/L) lyses the cells, and the cells are left to stand at a constant temperature of 80℃or 100℃for 2 hours until the cells are completely dissolved. The absorbance was measured at 405nm in a microplate reader. The melanin inhibition rate=1- (OD value per well/OD value of model group) ×100% was calculated.
As a result, as shown in FIG. 9, the melanin content of the blank group was 1, the melanin expression of the model group was 1.54, and the melanin inhibition rates of silymarin oil ceramide were 22.24%, 23.28%, 26.49%, 37.36%, 44.85% at concentrations of 10, 20, 40, 80, and 100mg/L, respectively, and excellent whitening effects were exhibited.
The foregoing is merely illustrative embodiments of the present invention, and the present invention is not limited thereto, and any changes or substitutions that may be easily contemplated by those skilled in the art within the scope of the present invention should be included in the scope of the present invention. Therefore, the protection scope of the present invention should be subject to the protection scope of the claims.

Claims (10)

1. Silybum oil ceramide, it is obtained by reacting fatty acid of Silybum oil with sphingosine compound, said sphingosine compound is selected from sphingosine, phytosphingosine, sphinganine.
2. Silybum marianum oil ceramide according to claim 1, wherein the silybum marianum oil fatty acid is obtained by hydrolyzing silybum marianum oil.
3. Silybum marianum oil ceramide according to claim 1 or 2, wherein the silybum marianum oil fatty acid contains 50-75wt% linoleic acid, 15-45wt% oleic acid, 1-10wt% palmitic acid, 1-6wt% stearic acid, 0.2-4wt% arachidic acid, 0.5-4wt% behenic acid and 0.01-2.5wt% linolenic acid.
4. Silybum marianum oil ceramide, which comprises the following components: linoleic acid ceramide, oleic acid ceramide, palmitic acid ceramide, stearic acid ceramide, arachidic acid ceramide, behenic acid ceramide, linolenic acid ceramide.
5. The silybum marianum oil ceramide of claim 4, which comprises the following components: 50 to 75 weight percent of linoleic acid ceramide, 15 to 45 weight percent of oleic acid ceramide, 1 to 10 weight percent of palmitic acid ceramide, 1 to 6 weight percent of stearic acid ceramide, 0.2 to 4 weight percent of arachidic acid ceramide, 0.5 to 4 weight percent of behenic acid ceramide and 0.01 to 2.5 weight percent of linolenic acid ceramide.
6. Silybum marianum oil ceramide according to claim 4 or 5, which further comprises: 0 to 1.5wt% of myristic acid ceramide.
7. The synthesis method of silybum marianum oil ceramide as claimed in any one of claims 1 to 6, comprising the following steps:
under the conditions of condensing agent and organic alkali, the silybum marianum oil fatty acid reacts with a sphingosine compound, wherein the condensing agent is EDCI, and the organic alkali is DIPEA;
the molar ratio of the silybum marianum oil fatty acid to the sphingosine compound to the EDCI to the DIPEA is 1: (1-1.5): (1-2): (1-2), wherein the solvent for the reaction is at least one of dichloromethane, tetrahydrofuran, ethyl acetate and acetonitrile.
8. Use of silybum marianum oil ceramide according to any one of claims 1-6 in cosmetics, pharmaceuticals, dietary or health care products.
9. The use according to claim 8, wherein the silybum marianum oil ceramide has at least one of skin barrier repair, tissue healing, anti-aging, anti-inflammatory, anti-photoaging, antioxidant, promoting collagen synthesis, maintaining elastin activity, whitening efficacy.
10. A composition comprising the silybum marianum oil ceramide of any one of claims 1-6, which has at least one of skin barrier repair, tissue healing, anti-aging, anti-inflammatory, anti-photoaging, anti-oxidation, promotion of collagen synthesis, maintenance of elastin activity, whitening efficacy.
CN202310051508.3A 2022-11-25 2023-02-02 Silybum oil ceramide and its synthesis method and application Pending CN117304056A (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
CN202310051508.3A CN117304056A (en) 2023-02-02 2023-02-02 Silybum oil ceramide and its synthesis method and application
PCT/CN2023/133524 WO2024109867A1 (en) 2022-11-25 2023-11-23 Vegetable oil ceramides, synthesis method therefor, and use thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202310051508.3A CN117304056A (en) 2023-02-02 2023-02-02 Silybum oil ceramide and its synthesis method and application

Publications (1)

Publication Number Publication Date
CN117304056A true CN117304056A (en) 2023-12-29

Family

ID=89236077

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202310051508.3A Pending CN117304056A (en) 2022-11-25 2023-02-02 Silybum oil ceramide and its synthesis method and application

Country Status (1)

Country Link
CN (1) CN117304056A (en)

Similar Documents

Publication Publication Date Title
CN115974714A (en) Shea butter ceramide, and its synthesis method and use
CN1905886B (en) A composition containing ginsenoside F1 or compound K for skin external application
CN115947666A (en) Rice bran oil ceramide and synthesis method and application thereof
JP4939766B2 (en) Basement membrane stabilizer
CN115894279A (en) Olive oil ceramide and synthesis method and application thereof
KR20150100288A (en) Cosmetic composition comprising the extract of crude drug fermentation using the black yeast
CN116023288A (en) Grape seed oil ceramide and synthesis method and application thereof
CN116041206A (en) Safflower seed oil ceramide and its synthesis method and use
KR100680584B1 (en) Hydroxybenzamide derivatives, the method for preparing thereof and the cosmetic composition containing the same
CN116003287A (en) Sunflower seed oil ceramide and synthesis method and application thereof
CN116003289A (en) Watermelon seed oil ceramide and synthesis method and application thereof
CN116003288A (en) Nanmei oleo ceramide and its synthesis method and use
CN116041205A (en) Borage oil ceramide and synthetic method and application thereof
CN116003286A (en) Prinsepia utilis royle oil ceramide and synthesis method and application thereof
CN116023289A (en) Avocado oil ceramide and synthesis method and application thereof
CN115974715A (en) Peony seed oil ceramide, and synthesis method and application thereof
CN115974716A (en) Cottonseed oil ceramide and synthesis method and application thereof
CN115947667A (en) Coconut oil ceramide and synthesis method and application thereof
CN117304056A (en) Silybum oil ceramide and its synthesis method and application
CN117304059A (en) Tomato seed oil ceramide and synthetic method and application thereof
CN117342970A (en) Wheat germ oil ceramide and synthesis method and application thereof
CN117326965A (en) Oenothera biennis oil ceramide and its synthesis method and use
CN117326967A (en) Pterocarpus santalinus seed oil ceramide and synthesis method and application thereof
CN117304055A (en) Perilla seed oil ceramide and synthesis method and application thereof
CN117304058A (en) Walnut oil ceramide and synthesis method and application thereof

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination