CN117304055A - Perilla seed oil ceramide and synthesis method and application thereof - Google Patents

Perilla seed oil ceramide and synthesis method and application thereof Download PDF

Info

Publication number
CN117304055A
CN117304055A CN202310051491.1A CN202310051491A CN117304055A CN 117304055 A CN117304055 A CN 117304055A CN 202310051491 A CN202310051491 A CN 202310051491A CN 117304055 A CN117304055 A CN 117304055A
Authority
CN
China
Prior art keywords
ceramide
seed oil
perilla seed
acid
weight percent
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
CN202310051491.1A
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 CN202310051491.1A priority Critical patent/CN117304055A/en
Priority to PCT/CN2023/133524 priority patent/WO2024109867A1/en
Publication of CN117304055A publication Critical patent/CN117304055A/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 perilla seed oil ceramide which is obtained by reacting perilla seed oil fatty acid with a sphingosine compound, wherein the sphingosine compound is selected from sphingosine, phytosphingosine and dihydrosphingosine. The perilla seed oil ceramide has excellent performances in the aspects of repairing natural skin barriers, anti-inflammatory, tissue healing, anti-aging and the like, and has wide application prospects in the fields of cosmetics, health-care products, biological medicines and the like.

Description

Perilla seed oil ceramide and synthesis method and application thereof
Technical Field
The invention belongs to the technical field of biological medicines, and particularly relates to perilla seed 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 perilla seed oil is a vegetable oil extracted from perilla seeds by ultrasonic auxiliary solvent extraction and the like, is light yellow oily liquid, and is basically transparent and has no pungent taste. The perilla seed oil contains abundant unsaturated fatty acid, is the currently known natural vegetable oil with the highest alpha-linolenic acid content, has the alpha-linolenic acid content of up to 70 weight percent and shares the name of deep sea fish oil on land. In addition, oleic acid, linoleic acid, stearic acid, arachidonic acid, gamma-tocopherol, phytosterol, polyphenol, flavone, vitamin E, etc. The data show that it can eliminate excessive free radical, reduce the production of allergy inducing matter obviously, and has powerful antioxidant, antisenescent and allergy inhibiting effects. In addition, in the medical field, the perilla seed oil has various physiological functions of promoting brain development, enhancing intelligence, improving memory, protecting vision, reducing blood fat, preventing cardiovascular diseases, resisting tumors and the like, and is also important for the intelligence and vision development of infants and the prevention of brain function decline and Alzheimer disease of middle-aged and elderly people.
Disclosure of Invention
The invention aims to provide ceramide synthesized by utilizing perilla seed oil fatty acid of plant origin.
Another object of the present invention is to provide a method for synthesizing perilla seed oil ceramide, which uses natural plant-derived and readily available perilla seed oil grease or perilla seed oil fatty acid as a raw material.
It is another object of the present invention to provide the use of perilla seed 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, the perilla seed oil ceramide is obtained by reacting a perilla seed 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 taking the perilla seed oil as a raw material, selecting a proper strain, and fermenting to obtain the perilla seed 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 perilla seed oil fatty acid is obtained by hydrolyzing perilla seed oil grease.
Further, the perilla seed oil fatty acid contains 65 to 90wt% of alpha-linolenic acid, preferably 65 to 78wt%.
Further, the perilla seed oil fatty acid contains 4 to 30wt% linoleic acid, preferably 10 to 23wt%.
Further, the perilla seed oil fatty acid contains 4 to 20wt% of oleic acid, preferably 10 to 15wt%.
Further, the perilla seed oil fatty acid contains 1 to 4wt% of stearic acid, preferably 1 to 3wt%.
In addition, the perilla seed oil fatty acid also contains 0 to 3.5 weight percent of palmitic acid and 0 to 0.6 weight percent of arachidonic acid.
The composition of the perilla seed oil fatty acid is as follows: 65 to 90 weight percent of alpha-linolenic acid, 4 to 30 weight percent of linoleic acid, 4 to 20 weight percent of oleic acid, 1 to 4 weight percent of stearic acid, 0 to 3.5 weight percent of palmitic acid and 0 to 0.6 weight percent of arachidonic acid; the preferred composition is: 65 to 78 weight percent of alpha-linolenic acid, 10 to 23 weight percent of linoleic acid, 10 to 15 weight percent of oleic acid, 1 to 3 weight percent of stearic acid, 0 to 3.5 weight percent of palmitic acid and 0 to 0.6 weight percent of arachidonic acid.
The main component of the perilla seed oil fatty acid is alpha-linolenic acid, other fatty acids comprise linoleic acid, oleic acid and stearic 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 palmitic acid and arachidonic acid are not necessarily contained, and are optional components or unnecessary components.
The purple perilla seed oil ceramide comprises the following components: alpha-linolenic acid ceramide, linoleic acid ceramide, oleic acid ceramide, stearic acid ceramide; because fatty acids all participate in the same reaction, the mass ratio of ceramide after the reaction is not greatly changed, so the composition of the ceramide of the perilla seed oil is similar to that of the fatty acid of the perilla seed oil, and the composition of the ceramide of the perilla seed oil is as follows: 65 to 90 weight percent of alpha-linolenic acid ceramide, 4 to 30 weight percent of linoleic acid ceramide, 4 to 20 weight percent of oleic acid ceramide and 1 to 4 weight percent of stearic acid ceramide; the preferred composition is: 65 to 78 weight percent of alpha-linolenic acid ceramide, 10 to 23 weight percent of linoleic acid ceramide, 10 to 15 weight percent of oleic acid ceramide and 1 to 3 weight percent of stearic acid ceramide. The content of each component is different due to the different content of each fatty acid in the perilla seed oil fatty acid or the grease. In addition, the perilla seed oil ceramide also comprises ceramide obtained by the reaction of one or more of palmitic acid and arachidonic acid and sphingosine compounds, namely 0 to 3.5 weight percent of palmitic acid ceramide and 0 to 0.6 weight percent of arachidonic acid ceramide. The perilla seed oil ceramide also includes compounds which exist in the fatty acid of the perilla seed oil but do not react with sphingosine compounds, such as gamma-tocopherol, phytosterol, polyphenol, flavone, vitamin E, and the like.
The purple perilla seed oil ceramide comprises the following components: alpha-linolenic acid ceramide, linoleic acid ceramide, oleic acid ceramide; the alpha-linolenic acid ceramide accounts for 65 to 90 weight percent, preferably 65 to 78 weight percent, the linoleic acid ceramide accounts for 4 to 30 weight percent, preferably 10 to 23 weight percent, and the oleic acid ceramide accounts for 4 to 20 weight percent, preferably 10 to 15 weight percent.
Further, the perilla seed oil ceramide comprises stearic acid ceramide, and the stearic acid ceramide accounts for 1-4wt%.
Further, the perilla seed oil ceramide comprises no more than 3.5wt% of palmitic acid ceramide and no more than 0.6wt% of arachidonic acid ceramide, especially 0.1 to 3.5wt% of palmitic acid ceramide and 0.1 to 0.6wt% of arachidonic acid ceramide.
The alpha-linolenic acid ceramide is obtained by condensation reaction of alpha-linolenic acid and sphingosine compounds, and comprises alpha-linolenic acid phytosphingosine ceramide, alpha-linolenic acid sphingosine ceramide and alpha-linolenic acid dihydro sphingosine 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; 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; palmitic acid ceramide, arachidonic acid ceramide, and the like.
In a second aspect of the invention, a method for synthesizing perilla seed oil ceramide comprises the following steps:
under the conditions of condensing agent and organic alkali, the perilla seed oil fatty acid reacts with sphingosine compound, the condensing agent is EDCI, and the organic alkali is Et 3 N。
Further, the perilla seed oil fatty acid, sphingosine compound, EDCI, et 3 The molar ratio of N 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 perilla seed oil purchased in the market is generally in the form of grease, and needs to be hydrolyzed into perilla seed oil fatty acid, so the method further comprises the following steps:
the perilla seed oil fat is hydrolyzed by saponification reaction to obtain the perilla seed oil fatty acid.
Further, the saponification reaction is hydrolysis of perilla seed oil grease in potassium hydroxide solution.
Further, the mass ratio of the perilla seed oil grease to the potassium hydroxide is 1: (1-2).
In a third aspect of the invention, the use of perilla seed oil ceramide in cosmetics, pharmaceutical products, dietary or health care products.
Further, the perilla seed 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 perilla seed 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 viability maintaining, 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 invention utilizes the excellent characteristics of the organic source perilla seed oil, and the unsaturated fatty acid which is rich in the perilla seed oil, especially up to 70 percent of alpha-linolenic acid, and the perilla seed oil ceramide which is prepared by the mild reaction of the sphingosine compound which is also naturally existing in the skin, has excellent performance in the aspects of repairing, antioxidation, anti-aging and the like of the natural barrier of the skin, and has wide application prospect in the fields of cosmetics, health care products, biological medicines and the like. In addition, the active ingredients such as alpha-linolenic acid-derived ceramide and the like contained in the perilla seed oil ceramide synthesized by the method have obvious price advantages compared with the products prepared from the expensive alpha-linolenic acid raw material.
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 of perilla seed 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 perilla seed 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 perilla seed oil also contains gamma-tocopherol, phytosterol, polyphenol, flavone, vitamin E and other components, and the nutrients have the effects of moistening skin, enhancing cell viability and the like, and the ceramide synthesized by the perilla seed oil has a synergistic effect with other active components contained in the perilla seed oil, so that the ceramide has a better effect compared with the ceramide compounded according to similar proportion.
3. The cost is lower. The method of the invention can rapidly obtain a plurality of ceramide compound compositions, and the vegetable source perilla seed oil grease or fatty acid sources thereof are wide in sources, easy to obtain commercially, lower in cost, more environment-friendly and economical, and different from the idea of mixing and compounding different single ceramides, the fatty acid with a single component has high raw material price (especially the price of the alpha-linolenic acid raw material is very expensive), and different ceramides are required to be produced respectively and then compounded, so that the preparation cost is increased.
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, et 3 N refers to triethylamine. 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 fatty acid of perilla seed oil and phytosphingosine
The first step: 40g of perilla seed oil grease is dissolved in 50mL 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 the water phase, and adding 100mL of saturated foodBrine is washed once, and the organic phase is added with anhydrous Na 2 SO 4 Drying, filtering and concentrating in vacuum to obtain 32g of perilla seed oil fatty acid.
And a second step of: the perilla seed oil fatty acid (50 mmol based on the main component fatty acid), EDCI (60 mmol) and Et 3 N (60 mmol) was added to a 250mL round bottom flask, followed by 100mL of dichloromethane, followed by stirring at room temperature for 1 hour, followed by phytosphingosine (60 mmol) added to the reaction system, and 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 the perilla seed 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: alpha-linolenic acid-phytosphingosine ceramide 8.3min, linoleic acid-phytosphingosine ceramide 9.5min, oleic acid-phytosphingosine ceramide 11.3min, stearic acid-phytosphingosine ceramide 13.9min.
The obtained product is analyzed by high performance liquid chromatography, the content of alpha-linolenic acid-phytosphingosine ceramide, linoleic acid-phytosphingosine ceramide, oleic acid-phytosphingosine ceramide and stearic acid-phytosphingosine ceramide is 78%, 12%, 6%, 2% in sequence, and the rest is other components, and the content is less.
Example 2
Synthesis of ceramide from fatty acid and sphingosine of perilla seed oil
The first step: 40g of perilla seed oil grease is dissolved in 50mL 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 pH of the reaction system to 3, adding 150mL ethyl acetate to extract water phase, adding 100mL saturated salt water to wash once, and adding water phasePhase added anhydrous Na 2 SO 4 Drying, filtering and concentrating in vacuum to obtain 32g of perilla seed oil fatty acid.
And a second step of: the perilla seed oil fatty acid (50 mmol based on the main component fatty acid), EDCI (75 mmol) and Et 3 N (75 mmol) was added to a 250mL round bottom flask, followed by 100mL of dichloromethane, followed by stirring at room temperature for 1 hour, followed by sphingosine (50 mmol) added to the reaction system, and 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 the perilla seed 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: alpha-linolenic acid-sphingosine ceramide 7.9min, linoleic acid-sphingosine ceramide 8.7min, oleic acid-sphingosine ceramide 10.2min, palmitic acid-sphingosine ceramide 10.4min, stearic acid-sphingosine ceramide 13.6min.
The obtained product is analyzed by high performance liquid chromatography, the content of alpha-linolenic acid-sphingosine ceramide, linoleic acid-sphingosine ceramide, oleic acid-sphingosine ceramide, stearic acid-sphingosine ceramide and palmitic acid-sphingosine ceramide is sequentially 70%, 15%, 7%, 2% and 2%, and the rest is other components, and the content is less.
Example 3
Synthesis of ceramide from fatty acid and sphinganine of perilla seed oil
The first step: 40g of perilla seed oil grease is dissolved in 50mL 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 the water phase, adding 100mL of saturated salt water to washOnce, the organic phase is added with anhydrous Na 2 SO 4 Drying, filtering and concentrating in vacuo gave 34g of perilla seed oil fatty acid.
And a second step of: the perilla seed oil fatty acid (50 mmol based on the main component fatty acid), EDCI (100 mmol) and Et 3 N (100 mmol) was added to a 250mL round bottom flask, followed by 100mL of dichloromethane, followed by stirring at room temperature for 1 hour, followed by addition of sphinganine (70 mmol) to the reaction system, stirring at room temperature, and detection by TLC 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 the perilla seed 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: alpha-linolenic acid-dihydro sphingosine ceramide 8.3min, linoleic acid-dihydro sphingosine ceramide 9.4min, oleic acid-dihydro sphingosine ceramide 11.1min, stearic acid-dihydro sphingosine ceramide 13.6min, and arachidonic acid-dihydro sphingosine ceramide 14.0min.
The obtained product is analyzed by high performance liquid chromatography, the content of alpha-linolenic acid-dihydrosphingosine ceramide, linoleic acid-dihydrosphingosine ceramide, oleic acid-dihydrosphingosine ceramide, stearic acid-dihydrosphingosine ceramide and arachidonic acid-dihydrosphingosine ceramide is 68%, 13%, 14%, 3%, 0.5% in sequence, and the rest is other components, and the content is 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, the medium was removed after further incubation for 24h, 100. Mu.L of thiazole blue (MTT) was added to each well, and the measurement at 450nmAbsorbance at, cell viability = a calculated Drug delivery hole /A Blank hole ×100％。
As shown in FIG. 1, the perilla seed oil ceramide has a promoting effect on cell viability, and the cell viability rates at the concentrations of 0.97657, 1.95313, 3.90625, 7.8125, 15.625, 31.25, 62.5 and 125mg/L are 121.42%, 129.10%, 132.97%, 129.16%, 120.44%, 122.89%, 111.23% and 118.53%, respectively.
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 promoted cell proliferation effect and tissue repair potential inferior to that of stachyose oil 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 perilla seed oil ceramide has better tissue healing capacity. The solvent control group had a healing rate of 48.35% after 24 hours, the perilla seed oil ceramide had a healing rate of 92.41% after 24 hours, and ceramide 3B had a healing rate of 59.32% after 24 hours. 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, the perilla seed oil ceramide has a good effect of inhibiting elastase at various concentrations, specifically, the inhibition rate of elastase at a concentration of 0.25g/L is 9.13%, the inhibition rate of elastase at a concentration of 0.5g/L is 14.33%, the inhibition rate of elastase at a concentration of 1.0g/L is 21.67%, and the inhibition rate of elastase at a concentration of 2.0g/L is 26.67%.
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 the perilla seed 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 0.89, 0.66, 0.59 and 0.25 times of that of an LPS model group respectively, and the dosage dependence is shown, so that the perilla seed 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 purple perilla seed oil ceramide was 16%, 25% and 49% relative to the model group at the concentrations of 125, 250 and 400 mg/L; the expression level of MMP1 in the negative control group was 1, the expression level in the model group was 2.33, and the inhibition ratio of MMP1 expression in the purple perilla seed oil ceramide at 125, 250 and 400mg/L was 30%, 41 and 54% with respect to the model group.
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 perilla seed 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 4.43%, 9.91%, 16.10%, 18.26% and 25.40%, 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 is discarded, and after washing with Phosphate Buffer (PBS) for one to two times, 1mL of NaOH solution (1 mol/L) containing 10% DMSO by mass fraction is added to lyse the cells, and the cells are kept at a constant temperature of 80℃or 100℃for 2 hours until the cells are completely lysed. 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 shown in fig. 9, the melanin content of the blank group was 1, the melanin expression of the model group was 1.51, and the melanin inhibition rates of perilla seed oil ceramide were 1.94%, 12.52%, 20.00% and 27.79% at concentrations of 10, 20, 40, 80 and 100mg/L, respectively, and excellent whitening effect was 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. The perilla seed oil ceramide is obtained by reacting perilla seed oil fatty acid with a sphingosine compound, wherein the sphingosine compound is selected from sphingosine, phytosphingosine and dihydrosphingosine.
2. The perilla seed oil ceramide of claim 1, wherein the perilla seed oil fatty acid is obtained by hydrolysis of perilla seed oil.
3. The perilla seed oil ceramide of claim 1 or 2, wherein the perilla seed oil fatty acid comprises 65 to 90wt% of alpha-linolenic acid, 4 to 30wt% of linoleic acid, 4 to 20wt% of oleic acid, and 1 to 4wt% of stearic acid.
4. The purple perilla seed oil ceramide comprises the following components: alpha-linolenic acid ceramide, linoleic acid ceramide, oleic acid ceramide, stearic acid ceramide.
5. The perilla seed oil ceramide of claim 4, which comprises the following components: 65 to 90 weight percent of linolenic acid ceramide, 4 to 30 weight percent of linoleic acid ceramide, 4 to 20 weight percent of oleic acid ceramide and 1 to 4 weight percent of stearic acid ceramide.
6. The perilla seed oil ceramide of claim 4 or 5, further comprising: 0 to 3.5 weight percent of palmitic acid ceramide and 0 to 0.6 weight percent of arachidonic acid ceramide.
7. The method for synthesizing perilla seed oil ceramide according to any one of claims 1 to 6, comprising the following steps:
under the conditions of condensing agent and organic alkali, the fatty acid of the perilla seed oil reacts with the sphingosine compound,the condensing agent is EDCI, and the organic base is Et 3 N；
The perilla seed oil fatty acid, sphingosine compound, EDCI, et 3 The molar ratio of N 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 the perilla seed oil ceramide of any one of claims 1 to 6 in cosmetics, medicines, dietary foods or health care products.
9. The use according to claim 8, wherein the perilla seed 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 perilla seed oil ceramide of any one of claims 1 to 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.
CN202310051491.1A 2022-11-25 2023-02-02 Perilla seed oil ceramide and synthesis method and application thereof Pending CN117304055A (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
CN202310051491.1A CN117304055A (en) 2023-02-02 2023-02-02 Perilla seed oil ceramide and synthesis method and application thereof
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
CN202310051491.1A CN117304055A (en) 2023-02-02 2023-02-02 Perilla seed oil ceramide and synthesis method and application thereof

Publications (1)

Publication Number Publication Date
CN117304055A true CN117304055A (en) 2023-12-29

Family

ID=89296026

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202310051491.1A Pending CN117304055A (en) 2022-11-25 2023-02-02 Perilla seed oil ceramide and synthesis method and application thereof

Country Status (1)

Country Link
CN (1) CN117304055A (en)

Similar Documents

Publication Publication Date Title
US20090170787A1 (en) Cosmetic composition containing hydrolysates of icariin
CN1905886B (en) A composition containing ginsenoside F1 or compound K for skin external application
CN115974714A (en) Shea butter ceramide, and its synthesis method and use
CN115894279A (en) Olive oil ceramide and synthesis method and application thereof
CN115947666A (en) Rice bran oil ceramide and synthesis method and application thereof
CN115304509A (en) Ceramide compound containing chain carboxylic acid and preparation method and application thereof
CN116003289A (en) Watermelon seed oil ceramide and synthesis method and application thereof
CN116023288A (en) Grape seed oil ceramide and synthesis method and application thereof
CN116003287A (en) Sunflower seed oil ceramide and synthesis method and application thereof
CN116041206A (en) Safflower seed oil ceramide and its synthesis method and use
CN116023289A (en) Avocado oil ceramide and synthesis method and application thereof
CN116003288A (en) Nanmei oleo ceramide and its synthesis method and use
CN115974715A (en) Peony seed oil ceramide, and synthesis method and application thereof
CN116003286A (en) Prinsepia utilis royle oil ceramide and synthesis method and application thereof
CN116041205A (en) Borage oil ceramide and synthetic 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
CN117304055A (en) Perilla seed oil ceramide and synthesis method and application thereof
JP6979919B2 (en) Human normal cell activator
CN117304059A (en) Tomato seed oil ceramide and synthetic method and application thereof
CN117304058A (en) Walnut oil ceramide and synthesis method and application thereof
CN117326965A (en) Oenothera biennis oil ceramide and its synthesis method and use
CN117285432A (en) Linseed oil ceramide and synthesis method and application thereof
CN115894278A (en) Linolenic acid-derived ceramide, and preparation method and application thereof
CN117304061A (en) Acer truncatum seed 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