Ceramides derived from amino acids, their synthesis method and use

Abstract

The present invention belongs to the technical field of biopharmaceuticals and discloses a ceramide derived from an amino acid, a compound having the structure of general formula I, or an isomer thereof. is JPEG2026513487000069.jpg58170, and the said R 27 , 14 , 29 , 14 , 27 is a residue to which a polypeptide is condensed, and the said R 3 is selected from alanine, threonine, proline, asparagine, glutamine, leucine, tryptophan, serine, valine, methionine, tyrosine, histidine, L-aspartic acid-4-tert-butyl ester, L-aspartic acid-1-tert-butyl ester, glycine, isoleucine, phenylalanine, lysine, arginine, cysteine, L-glutamic acid-5-tert-butyl ester, L-glutamic acid-1-tert-butyl ester, and a residue to which a polypeptide is condensed. The said polypeptide is one formed by condensation of 2 to 10 amino acids. R 2 is -C 15 H 29 、-C 15 H 31 、-C 15 H 27 、-CHOHC 14 H 27 、-CHOHC 14 H 29 is selected from. The present invention further discloses a synthesis method of such a compound. Such a compound has excellent effects in moisturizing, antioxidation, anti-glycation, skin barrier repair, tissue healing, etc.

Description

Technical Field

[0001] The present invention belongs to the technical field of biopharmaceuticals, and specifically relates to ceramides derived from amino acids, their synthesis methods and uses.

Background Art

[0002] Ceramide (also called molecular nail) naturally exists in the skin and is a very important component of the skin barrier (stratum corneum), and its content reaches 40 - 50 wt%. Ceramide is a sphingolipid composed of a sphingosine-based long-chain base and a fatty acid. The sphingosine part, the carbon chain length, the degree of unsaturation and the number of hydroxyl groups of the fatty acid part vary, and ceramide represents a compound. Ceramide exhibits excellent performance in terms of regulating skin barrier function, restoring skin moisture, and enhancing the adhesion force between skin keratinocytes.

[0003] When the amount of ceramide decreases, it causes skin dryness, loses the defense function of the skin surface, makes foreign substances more likely to invade, causes secondary skin infections, and causes skin rejection reactions. Specifically, the invaders cause cytokines to be released from cells such as keratinocytes, Langerhans cells, and melanocytes of the surface cells, causing inflammatory phenomena and the like. Therefore, in order to maintain and improve the skin barrier, skin moisturization is important. A physiological lipid mixture containing ceramide-based compounds can promote the recovery of the damaged skin barrier function compared with general moisturizers. As a result of clinical trials, it showed the same effect as medium to high-grade topical steroid preparations in improving the symptoms of atopic dermatitis patients.

[0004] Due to the importance of ceramide, many cosmetic and pharmaceutical companies are researching and developing corresponding products. However, it is difficult to mass-produce natural ceramide because it is difficult to extract and has a high cost and is not suitable for commercialization. There are also companies working on the development of ceramides that have a similar structure to the ceramide present in the skin and can provide functionally similar effects.

[0005] Therefore, considering the broad market demand for functional ceramides, it is necessary to address the supply shortage and enhance their effectiveness by constructing novel ceramide compounds using naturally derived and readily available raw materials. [Overview of the project]

[0006] The object of the present invention is to provide a ceramide derived from an amino acid with a novel structure. This is produced by the bonding of an amino acid or polypeptide with a sphingoid base compound or a sphingoid base compound to which succinic acid is bonded.

[0007] Another object of the present invention is to provide a method for synthesizing ceramides derived from amino acids. This method uses an amino acid, a polypeptide, and a sphingoid base compound or a sphingoid base compound to which succinic acid is bonded as raw materials.

[0008] Another object of the present invention is to provide the use of ceramides derived from amino acids.

[0009] To achieve one of the above objectives, the present invention employs the following technical solutions. A ceramide derived from an amino acid, having a structure of general formula I, or an isomer thereof. In the JPEG2026513487000002.jpg formula, M is R 3 or The file is JPEG2026513487000003.jpg20170, and the aforementioned R 1 These are residues formed by the condensation of polypeptides, The aforementioned R 3is selected from residues condensed from alanine, threonine, proline, asparagine, glutamine, leucine, tryptophan, serine, valine, methionine, tyrosine, histidine, L-aspartic acid-4-tert-butyl ester, L-aspartic acid-1-tert-butyl ester, glycine, isoleucine, phenylalanine, lysine, arginine, cysteine, L-glutamic acid-5-tert-butyl ester, L-glutamic acid-1-tert-butyl ester, and residues of condensed polypeptides, the polypeptide is one formed by condensation of 2 to 10 amino acids, R 2 is -C 15 H 29 、-C 15 H 31 、-C 15 H 27 、-CHOHC 14 H 27 、-CHOHC 14 H 29 and is one selected therefrom.

[0010] The condensed residue means that after the carboxy group of the corresponding amino acid RCOOH and the amino group of the sphingoid base are condensed to form a peptide bond, the remaining amino acid fragment R, for example, alanine The residue condensed with JPEG2026513487000004.jpg14170 is JPEG2026513487000005.jpg8170, and for threonine JPEG2026513487000006.jpg19170JPEG2026513487000007.jpg18170. Or, the carboxy group of the corresponding polypeptide and the amino group of the sphingoid base are condensed to form a peptide bond, or the amino group of the polypeptide and the sphingoid base to which succinic acid is bonded The residue condensed with JPEG2026513487000008.jpg40170JPEG2026513487000009.jpg17170 is The image is JPEG2026513487000010.jpg15170, and the residue formed by the condensation of the tetrapeptide-5-H2N-(β-Ala)-His-Ser-His-OH is (β-Ala)-His-Ser-His-OH.

[0011] Furthermore, the R 3 The residues are selected from those formed by the condensation of alanine, threonine, proline, asparagine, glutamine, leucine, tryptophan, serine, valine, methionine, tyrosine, histidine, L-aspartate-4-tert-butyl ester, L-aspartate-1-tert-butyl ester, glycine, isoleucine, phenylalanine, lysine, arginine, cysteine, L-glutamate-5-tert-butyl ester, L-glutamate-1-tert-butyl ester, dipeptides, tetrapeptides, hexapeptides, octapeptides, and nonapeptides.

[0012] Furthermore, the R 3 The residues are selected from those formed by the condensation of alanine, threonine, proline, asparagine, glutamine, leucine, tryptophan, serine, valine, methionine, tyrosine, histidine, L-aspartate-4-tert-butyl ester, L-aspartate-1-tert-butyl ester, glycylglycine, snake venom peptide intermediate, alanyl-L-tyrosine, tetrapeptide-5, hexapeptide-1, hexapeptide-8, hexapeptide-9, octapeptide-3, and nonapeptide-1.

[0013] Furthermore, the R 3 The residues are selected from alanine, threonine, proline, asparagine, glutamine, leucine, tryptophan, serine, valine, methionine, tyrosine, histidine, L-aspartate-4-tert-butyl ester, L-aspartate-1-tert-butyl ester, glycylglycine, snake venom peptide intermediates, and residues formed by the condensation of alanyl-L-tyrosine.

[0014] Furthermore, the R 3The residues are selected from those formed by the condensation of alanine, threonine, proline, asparagine, glutamine, leucine, tryptophan, serine, valine, methionine, tyrosine, histidine, L-aspartate-4-tert-butyl ester, L-aspartate-1-tert-butyl ester, glycine, isoleucine, phenylalanine, lysine, arginine, cysteine, L-glutamate-5-tert-butyl ester, and L-glutamate-1-tert-butyl ester.

[0015] Furthermore, the R 3 The residues are selected from those formed by the condensation of alanine, threonine, proline, asparagine, glutamine, leucine, tryptophan, serine, valine, methionine, tyrosine, histidine, L-aspartate-4-tert-butyl ester, and L-aspartate-1-tert-butyl ester.

[0016] Furthermore, the R 3 These are residues formed by the condensation of polypeptides.

[0017] Furthermore, the R 3 The residue is selected from the condensed remains of dipeptides, tetrapeptides, hexapeptides, octapeptides, and nonapeptides.

[0018] A dipeptide is a polypeptide in which two amino acids are linked by one peptide bond. A tetrapeptide is a polypeptide in which four amino acids are linked by three peptide bonds. A hexapeptide is a polypeptide in which six amino acids are linked by five peptide bonds. An octapeptide is a polypeptide in which eight amino acids are linked by seven peptide bonds. A nonapeptide is a polypeptide in which nine amino acids are linked by eight peptide bonds.

[0019] Furthermore, the R 3The residue is selected from the condensed residues of glycylglycine, snake venom peptide intermediate, alanyl-L-tyrosine, tetrapeptide-5, hexapeptide-1, hexapeptide-8, hexapeptide-9, octapeptide-3, and nonapeptide-1.

[0020] Furthermore, the R 3 This is selected from residues formed by the condensation of glycylglycine, a snake venom peptide intermediate, and alanyl-L-tyrosine.

[0021] Furthermore, the R 1 These are residues formed by the condensation of polypeptides.

[0022] Furthermore, the R 1 The residue is selected from the condensed remains of dipeptides, tetrapeptides, hexapeptides, octapeptides, and nonapeptides.

[0023] Furthermore, the R 1 The residue is selected from the condensed residues of glycylglycine, snake venom peptide intermediate, alanyl-L-tyrosine, tetrapeptide-5, hexapeptide-1, hexapeptide-8, hexapeptide-9, octapeptide-3, and nonapeptide-1.

[0024] Furthermore, the R 1 The residue is selected from the condensed residues of tetrapeptide-5, hexapeptide-1, hexapeptide-8, hexapeptide-9, octapeptide-3, and nonapeptide-1.

[0025] Composition R 3 or R 1 The amino acids of the polypeptide are selected from alanine, threonine, proline, asparagine, glutamine, leucine, tryptophan, serine, valine, methionine, tyrosine, histidine, L-aspartate-4-tert-butyl ester, L-aspartate-1-tert-butyl ester, glycine, isoleucine, phenylalanine, lysine, arginine, cysteine, L-glutamate-5-tert-butyl ester, and L-glutamate-1-tert-butyl ester.

[0026] Structural formula of glycylglycine: JPEG2026513487000011.jpg18170

[0027] Structural formula of snake venom peptide intermediate (dipeptide-1): JPEG2026513487000012.jpg20170

[0028] Structural formula of alanyl-L-tyrosine (dipeptide-2): JPEG2026513487000013.jpg29170

[0029] Structural formula of tetrapeptide-5: H-(b-Ala)-His-Ser-His-OH

[0030] Structural formula of hexapeptide-1: H-Nle-Ala-His-(D-Phe)-Arg-Trp-NH2

[0031] Structural formula of hexapeptide-8: H-Glu-Glu-Met-Gln-Arg-Arg-NH2

[0032] Structural formula of hexapeptide-9: H-Gly-Pro-Gln-Gly-Pro-Gln-OH

[0033] Structural formula of octapeptide-3: H-Glu-Glu-Met-Gln-Arg-Arg-Ala-Asp-NH2

[0034] Structural formula of nonapeptide-1: H-Met-Pro-D-Phe-Arg-D-Trp-Phe-Lys-Pro-Val-NH2

[0035] Furthermore, the R 2 It is one of the following structures: JPEG2026513487000014.jpg118170

[0036] Furthermore, the R 2It is one of the following structures: JPEG2026513487000015.jpg32170 corresponds to sphingosine, dihydrosphingosine, and phytosphingosine, respectively.

[0037] Furthermore, the R 3 This is a residue formed by the condensation of alanine.

[0038] Furthermore, the R 3 This is a residue formed by the condensation of threonine.

[0039] Furthermore, the R 3 This is a residue formed by the condensation of proline.

[0040] Furthermore, the R 3 This is a residue formed by the condensation of asparagine.

[0041] Furthermore, the R 3 This is a residue formed by the condensation of glutamine molecules.

[0042] Furthermore, the R 3 This is a residue formed by the condensation of leucine.

[0043] Furthermore, the R 3 This is a residue formed by the condensation of tryptophan.

[0044] Furthermore, the R 3 This is a residue formed by the condensation of serine.

[0045] Furthermore, the R 3 This is a residue formed by the condensation of valine.

[0046] Furthermore, the R 3 This is a residue formed by the condensation of methionine.

[0047] Furthermore, the R 3 This is a residue formed by the condensation of tyrosine molecules.

[0048] Furthermore, the R 3This is a residue formed by the condensation of histidine molecules.

[0049] Furthermore, the R 3 This is a residue obtained by condensing L-aspartate-4-tert-butyl ester or L-aspartate-1-tert-butyl ester.

[0050] Furthermore, the R 3 This is a residue formed by the condensation of glycylglycine.

[0051] Furthermore, the R 3 This is a residue formed by the condensation of a snake venom peptide intermediate.

[0052] Furthermore, the R 3 This is a residue formed by the condensation of alanyl-L-tyrosine.

[0053] Furthermore, the R 3 This is a residue formed by the condensation of tetrapeptide-5.

[0054] Furthermore, the R 3 This is a residue formed by the condensation of hexapeptide-1.

[0055] Furthermore, the R 3 This is a residue formed by the condensation of hexapeptide-8.

[0056] Furthermore, the R 3 This is a residue formed by the condensation of hexapeptide-9.

[0057] Furthermore, the R 3 This is a residue formed by the condensation of octapeptide-3.

[0058] Furthermore, the R 3 This is a residue formed by the condensation of nonapeptide-1.

[0059] Furthermore, the R 1 This is a residue formed by the condensation of glycylglycine.

[0060] Furthermore, the R 1This is a residue formed by the condensation of a snake venom peptide intermediate.

[0061] Furthermore, the R 1 This is a residue formed by the condensation of alanyl-L-tyrosine.

[0062] Furthermore, the R 1 This is a residue formed by the condensation of tetrapeptide-5.

[0063] Furthermore, the R 1 This is a residue formed by the condensation of hexapeptide-1.

[0064] Furthermore, the R 1 This is a residue formed by the condensation of hexapeptide-8.

[0065] Furthermore, the R 1 This is a residue formed by the condensation of hexapeptide-9.

[0066] Furthermore, the R 1 This is a residue formed by the condensation of octapeptide-3.

[0067] Furthermore, the R 1 This is a residue formed by the condensation of nonapeptide-1.

[0068] Furthermore, ceramide derived from amino acids is one of the following compounds to be selected. JPEG2026513487000016.jpg110170JPEG2026513487000017.jpg110170

[0069] The method for synthesizing ceramides derived from amino acids includes the following steps. M is R 3 If that is the case, Step S1: Boc or Fmoc-protected polypeptide or amino acid, sphingoid base Compound C is obtained by reacting JPEG2026513487000018.jpg18170, a condensing agent, and a coupling agent. Step S2: The protecting group Boc or Fmoc is removed from compound C to obtain the product. M In the case of JPEG2026513487000019.jpg20170, the protecting group Fmoc is removed from the Fmoc-protected polypeptide, and then a sphingoid base is bonded to succinic acid. JPEG2026513487000020.jpg37170, reacted with a condensing agent to obtain the product.

[0070] M is R 3 In this case, the condensing agent is EDCI or DCC, the coupling agent is N-hydroxysuccinimide, the molar ratio of the Boc or Fmoc-protected amino acid or polypeptide, sphingoid base, condensing agent, and coupling agent is (1-1.5):1:(1-2):(1-2), and the solvent for the S1 reaction is dichloromethane.

[0071] M In the case of JPEG2026513487000021.jpg20170, the condensing agent is HOBt and DIC, and the polypeptide and succinic acid are bonded to a sphingoid base. The molar ratio of JPEG2026513487000022.jpg37170, HOBt, and DIC is (1~1.5):1:(1~2):(1~2), and the solvent for the reaction is DMF.

[0072] Step S1 specifically involves dissolving Boc or Fmoc-protected amino acids or polypeptides, a coupling agent (EDCI or DIC), and N-hydroxysuccinimide in dichloromethane, reacting them at room temperature for a specified time, adding a sphingoid base, and continuing to react with stirring at room temperature until the sphingoid base has completely reacted, as confirmed by TLC detection. In the workup, the mixture is diluted with water or DCM, separated, filtered to remove the solid, the organic phases are washed with water and saturated brine respectively, the aqueous phase is further extracted with dichloromethane, the organic phases are combined, dried over anhydrous sodium sulfate, filtered, and then evaporated under reduced pressure to obtain the crude product, which is then purified by column chromatography to obtain the final product.

[0073] R 3 If the amino acid is present, in S2, for the Boc protecting group, the crude product is dissolved in MeOH, 6N hydrochloric acid is slowly added dropwise, and the mixture is heated to 50°C and allowed to react until TLC detection confirms that the crude product starting material has reacted completely. In the workup, MeOH is removed by rotary evaporation, the pH of the reaction solution is adjusted to approximately 9 with saturated sodium carbonate solution, diluted with saturated brine, further extracted with dichloromethane, the resulting organic phase is collected, dried over anhydrous sodium sulfate, filtered, and subjected to reduced-pressure rotary evaporation to obtain the crude product.

[0074] For the Fmoc protecting group, the crude product is dissolved in THF, diethylamine is slowly added dropwise, and the mixture is allowed to react at room temperature until the crude product is confirmed to be completely reacted by TLC detection. Workup: THF is removed by rotary evaporation, the mixture is diluted with saturated brine, and further extracted with dichloromethane. The resulting organic phase is collected, dried over anhydrous sodium sulfate, filtered, and subjected to reduced-pressure rotary evaporation to obtain the crude product.

[0075] A method for synthesizing ceramide derived from amino acids further comprises step S3. S3: The crude product is recrystallized with methanol, and the product is obtained by filtration, or the crude product is purified by column chromatography to obtain the product.

[0076] R 3 If the product is a polypeptide, S2 is specifically as follows. The product obtained in the above steps is dissolved in DCM, TFA is added, and the mixture is reacted overnight at room temperature with stirring until it is confirmed by TLC detection that the starting materials have reacted completely. Workup: After removing the reaction mixture by rotary evaporation, saturated sodium carbonate solution is added to adjust the pH of the reaction system to about 10, DCM is added and then separated, the organic phase is washed twice with water, the combined aqueous phase is extracted once with DCM, the organic phases are combined, dried over anhydrous sodium sulfate, filtered, and then the crude product is obtained by rotary evaporation under reduced pressure, the crude product is recrystallized with methanol, and then filtered to obtain a purified white solid product.

[0077] M If the filename is JPEG2026513487000023.jpg20170, the specific steps are as follows: After synthesizing the polypeptide in the resin, piperidine is added to remove the Fmoc protecting group of the polypeptide. JPEG2026513487000024.jpg37170 and HOBt are weighed, dissolved in DMF solvent, activated with DIC under an ice bath, then introduced into a reaction column and reacted at room temperature. After washing the resin with DCM, DMF, and methanol, the resulting crude peptide resin is dried in a vacuum drying oven. Decomposition and separation: The dried resin is placed in a decomposition solution (TFA:EDT:TIPS:H2O = 92.5:2.5:2.5:2.5), decomposed at room temperature, and precipitated with frozen isopropyl ether. After a white solid precipitate is formed, the white solid is filtered by suction and dried, dissolved in water and acetonitrile, and the product is separated using a half-separation apparatus under a gradient of 30-70 (mobile phase: 1 / ‰ trifluoroacetic acid aqueous solution and acetonitrile).

[0078] M If the filename is JPEG2026513487000025.jpg20170, the synthesis method is sphingoid base Steps to react JPEG2026513487000026.jpg19170 with succinic anhydride: Further includes JPEG2026513487000027.jpg39170.

[0079] The organic base DIPEA is added to this reaction, tetrahydrofuran THF is used as the solvent, and the molar ratio of the sphingoid base, succinic anhydride, and DIPEA is 1:(1~1.5):(1.5~3). The specific steps are as follows: Succinic anhydride and sphingosine are added to a necked flask, THF is added to dissolve them, then DIPEA is added, and the reaction is carried out at room temperature with stirring until complete reaction is confirmed by TLC detection. Extraction is performed twice with dichloromethane and hydrochloric acid aqueous solution, and once with dichloromethane and saturated brine solution. The organic phases are combined and purified by column chromatography to obtain a white solid product.

[0080] Ceramides derived from amino acids possess at least one of the following functions: skin barrier repair, tissue healing, antioxidant, anti-glycation, and moisturizing, and can be applied to cosmetics, health foods, pharmaceuticals, and especially cosmetic essence oils or anhydrous formulations.

[0081] Further provided are compositions containing ceramide derived from amino acids, its isomers, pharmaceutically acceptable salts thereof, its hydrates, or its solvates as active ingredients. These compositions have skin barrier repair, tissue healing, antioxidant, anti-glycation, or moisturizing effects.

[0082] This composition contains acceptable excipients. The acceptable excipients include one or more of the following: solubilizers, preservatives, antioxidants, pH adjusters, penetration enhancers, liposomes, humectants, thickeners, chelating agents, texture modifiers, surfactants, emulsifiers, fragrances, and colorants. This composition is available in the form of a cream, emulsion, solution, film, aerosol, or spray.

[0083] As used herein, the term "isomer" includes tautomers and stereoisomers. Tautomers are structural isomers that have different energies and can be converted into each other by low energy barriers, while stereoisomers are compounds that have the same chemical structure but differ in the spatial arrangement of atoms or groups. Stereoiomers include enantiomers, diastereomers, conformational isomers (rotational isomers), geometric isomers (cis / trans), and stereoisomers.

[0084] As used herein, “pharmaceutically acceptable salt” means a salt according to one embodiment of the present invention that is pharmaceutically acceptable and has the desired pharmacological activity of the parent compound. Such salts include (1) an acid addition salt with an inorganic or organic acid, or (2) a salt in which an acidic proton present in the parent compound is substituted. Examples of inorganic acids include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. Examples of organic acids include acetic acid, propionic acid, caproic acid, cyclopentylpropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, hydroxysuccinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid, 2-isethionic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo[2,2,2]-octa-2-ene-1-carboxylic acid, glucoheptonic acid, 3-benzenepropionic acid, trimethylacetic acid, tert-butylacetic acid, dodecyl sulfate, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, and muconic acid.

[0085] As used herein, "hydrate" refers to a compound bonded with water. The bond between the compound and water may include non-covalent bonds.

[0086] As used herein, "solvate" refers to a complex formed by a solute molecule or ion and a solvent molecule or ion.

[0087] Unless otherwise specified, the term "compound of the present invention" includes the compound itself, its pharmaceutically acceptable salts, its hydrates, its solvates, and its isomers.

[0088] The present invention has the following beneficial effects. In this invention, amino acids or polypeptides are used as starting materials and reacted with sphingoid bases having long-chain alkyl groups to construct ceramides derived from amino acids with novel structures. Such compounds improve the lipid solubility of amino acid-based compounds or polypeptides, are advantageous for skin penetration, and exhibit excellent effects in antioxidant, anti-glycation, skin barrier repair, tissue healing, and moisturizing properties, making them applicable to health foods, cosmetics, and pharmaceuticals. The improved lipid solubility of the compounds makes them more suitable for use in oil-based skincare formulations, solving the problem that unmodified peptides cannot be used in oil-phase systems. [Brief explanation of the drawing]

[0089] [Figure 1] This is a histogram of the antioxidant experiment results for Example 30. [Figure 2] This is a histogram of the antioxidant experiment results for Example 30. [Figure 3] This is a histogram of the antioxidant experiment results for Example 30. [Figure 4] This is a histogram of the antioxidant experiment results for Example 30. [Figure 5] This is a histogram of the antioxidant experiment results for Example 30. [Figure 6] This is a histogram of the antioxidant experiment results for Example 30. [Figure 7] This is a histogram of the antioxidant experiment results for Example 30. [Figure 8] This is a histogram of the antioxidant experiment results for Example 30. [Figure 9] This is a histogram of the antioxidant experiment results for Example 30. [Figure 10] This is a histogram of the antioxidant experiment results for Example 30. [Figure 11] This is a histogram of the antioxidant experiment results for Example 30. [Figure 12] This is a histogram of the antioxidant experiment results for Example 30. [Figure 13] This is a histogram of the anti-glycation experiment results for Example 31. [Figure 14]This is a histogram of the anti-glycation experiment results for Example 31. [Figure 15] This is a histogram of the anti-glycation experiment results for Example 31. [Figure 16] This is the mass spectrum of the compound from Example 20. [Figure 17] This is the mass spectrum of the compound from Example 21. [Figure 18] This is the mass spectrum of the compound from Example 22. [Figure 19] This is the mass spectrum of the compound from Example 23. [Figure 20] This is the mass spectrum of the compound from Example 24. [Figure 21] This is the mass spectrum of the compound in Example 25. [Figure 22] This is the mass spectrum of the compound from Example 26. [Figure 23] This is the mass spectrum of the compound from Example 27. [Figure 24] This is the mass spectrum of the compound from Example 28. [Figure 25] This is the mass spectrum of the compound from Example 29. [Figure 26] This shows the moisture absorption rate-time curves for hyaluronic acid, glycylglycine-phytosphingosine, snake venom peptide-phytosphingosine, and dipeptide-2-phytosphingosine in an 81% ambient humidity environment in Example 32. [Figure 27] This is the moisture absorption rate-time curve for tetrapeptide-5-phytosphingosine, tetrapeptide-5-sphingosine, hexapeptide-1-phytosphingosine, and hexapeptide-1-sphingosine at 81% ambient humidity in Example 32. [Figure 28] This shows the moisture absorption rate-time curves for hexapeptide-8-phytosphingosine, hexapeptide-9-phytosphingosine, hexapeptide-9-sphingosine, and octapeptide-3-phytosphingosine at 81% ambient humidity in Example 32. [Figure 29]These are the moisture absorption rate-time curves for nonapeptide-1-phytosphingosine, nonapeptide-1-sphingosine, and hyaluronic acid, glycylglycine-phytosphingosine at 81% ambient humidity in Example 32, and at 43% ambient humidity. [Figure 30] This shows the moisture absorption rate-time curves for snake venom peptide-phytosphingosine, dipeptide-2-phytosphingosine, tetrapeptide-5-phytosphingosine, and tetrapeptide-5-sphingosine in a 43% ambient humidity environment in Example 32. [Figure 31] This shows the moisture absorption rate-time curves for hexapeptide-1-phytosphingosine, hexapeptide-1-sphingosine, hexapeptide-8-phytosphingosine, and hexapeptide-9-phytosphingosine at 43% ambient humidity in Example 32. [Figure 32] This shows the moisture absorption rate-time curves for hexapeptide-9-sphingosine, octapeptide-3-phytosphingosine, nonapeptide-1-phytosphingosine, and nonapeptide-1-sphingosine in a 43% ambient humidity environment in Example 32. [Modes for carrying out the invention]

[0090] The present invention will be further described below with reference to specific examples.

[0091] All reactions were carried out under a nitrogen atmosphere. Unless otherwise specified, chemicals were purchased commercially and not further purified. The dichloromethane, tetrahydrofuran, pyridine, and N,N-dimethylformamide used in the experiments were all anhydrous solvents. Thin-layer chromatography (TLC) analysis was performed using 60F254 silica gel plates. Qingdao marine silica gel (particle size 0.040-0.063 mm) was used for silica gel column chromatography. Spot detection in TLC was performed using UV light (254 nm) or iodine. NMR spectra were obtained using a Bruker DPX 400 nuclear magnetic resonance instrument. 1The 1H NMR spectrum was measured at 400 MHz, using deuterated methanol, deuterated DMSO, or deuterated tetrahydrofuran as the solvent, with tetramethylsilane (TMS) as the internal standard. Chemical shifts were measured in ppm, and bond constants were measured in Hz. 1 In 1H NMR, δ represents the chemical shift, s represents a single peak, d represents a double peak, t represents a triple peak, q represents a quadruple peak, and m represents a multi-peak peak.

[0092] EDCI refers to 1-ethyl-(3-dimethylaminopropyl)carbodiimide, HOBt to 1-hydroxybenzotriazole, DCC to N'N-dicyclohexylcarbodiimide, DIC to N'N-diisopropylcarbodiimide, SuOH to N-hydroxysuccinimide, DCM to dichloromethane, THF to tetrahydrofuran, DMF to N,N-dimethylformamide, Boc to tert-butoxycarbonyl, Fmoc to 9-fluorenylmethoxycarbonyl, TFA to trifluoroacetic acid, EA to ethyl acetate, MeOH to methanol, and DIPEA to N,N-diisopropylethylamine.

[0093] Method A Condensation reaction: Boc-protected amino acids (1.0 eq, 50 mmol), EDCI (1.5 eq, 75 mmol), and N-hydroxysuccinimide (1.5 eq, 75 mmol) were dissolved in 100 mL of dichloromethane and reacted at room temperature for 30 minutes. Then, phytosphingosine (1 eq, 50 mmol) was added, and the reaction was continued with stirring at room temperature until complete reaction of phytosphingosine was confirmed by TLC detection.

[0094] The dose of Boc-protected amino acids may be adjusted to 50-75 mmol (1-1.5 eq.), the doses of EDCI and N-hydroxysuccinimide may be adjusted to 50-100 mmol (1-2 eq.), and sphingosine or dihydrosphingosine may be used instead of phytosphingosine.

[0095] Workup: The solution was diluted with 50 mL of water, separated, and the organic phase was washed once each with 100 mL of water and 100 mL of saturated saline solution. The aqueous phase was extracted once with 60 mL of dichloromethane, the organic phase was combined, dried over anhydrous sodium sulfate, filtered, and the crude product was obtained by rotary evaporation under reduced pressure.

[0096] Deprotection reaction: The crude product (1 eq, 30 mmol) was dissolved in 100 mL of MeOH, and 6N hydrochloric acid (4 eq, 120 mmol) was slowly added dropwise. The mixture was heated to 50°C and allowed to react until the crude product starting material had completely reacted, as confirmed by TLC detection.

[0097] Workup: Remove MeOH by rotary evaporation and adjust the pH of the reaction solution to approximately 9 with saturated sodium carbonate solution. Dilute with 100 mL of saturated saline solution, extract with 80 mL of dichloromethane, collect the resulting organic phase, dry with anhydrous sodium sulfate, filter, and obtain the crude product by reduced-pressure rotary evaporation.

[0098] Purification process of the crude product: The crude product was recrystallized with methanol, and then filtered to obtain a purified white solid product.

[0099] Method B Condensation reaction: Boc-protected amino acids (1.3 eq, 50 mmol) and N-hydroxysuccinimide (1.5 eq, 75 mmol) were dissolved in 100 mL of dichloromethane. DIC (1.5 eq, 75 mmol) was added dropwise at room temperature, and the mixture was allowed to react overnight at room temperature. Then, phytosphingosine (1 eq, 50 mmol) was added, and the mixture was stirred at room temperature until TLC detection confirmed that phytosphingosine had completely reacted.

[0100] The dose of Boc-protected amino acids may be adjusted to 50-75 mmol (1-1.5 eq.), the doses of DIC and N-hydroxysuccinimide may be adjusted to 50-100 mmol (1-2 eq.), and sphingosine or dihydrosphingosine may be used instead of phytosphingosine.

[0101] Workup: The solution was diluted with 80 mL of water, separated, and the organic phase was washed once each with 80 mL of water and 80 mL of saturated saline solution. The aqueous phase was extracted once with 80 mL of dichloromethane, the organic phase was combined, dried over anhydrous sodium sulfate, filtered, and the crude product was obtained by rotational evaporation under reduced pressure. The product was then purified by column chromatography to obtain the final product.

[0102] Deprotection reaction: The crude product (1 eq, 30 mmol) was dissolved in 100 mL of MeOH, and 6N hydrochloric acid (4 eq, 120 mmol) was slowly added dropwise. The mixture was heated to 50°C and allowed to react until the crude product was confirmed to be completely reacted by TLC detection.

[0103] Workup: Remove MeOH by rotary evaporation and adjust the pH of the reaction solution to approximately 9 with saturated sodium carbonate solution. Dilute with 100 mL of saturated saline solution, then extract with 80 mL of dichloromethane. Collect the resulting organic phase, dry with anhydrous sodium sulfate, filter, and obtain the crude product by reduced-pressure rotary evaporation.

[0104] Purification process of the crude product: The crude product was recrystallized with methanol and filtered to obtain a white solid product. This product was then slurryed with dichloromethane and filtered to obtain a purified product.

[0105] Method C Condensation reaction: Fmoc-protected amino acids (1.3 eq, 50 mmol) and N-hydroxysuccinimide (1.5 eq, 75 mmol) were dissolved in 100 mL of dichloromethane. DIC (1.5 eq, 75 mmol) was added dropwise at room temperature, and the mixture was allowed to react overnight at room temperature. Then, phytosphingosine (1 eq, 50 mmol) was added, and the mixture was stirred at room temperature until TLC detection confirmed that phytosphingosine had completely reacted.

[0106] The dose of Fmoc-protected amino acids may be adjusted to 50-75 mmol (1-1.5 eq.), the doses of DIC and N-hydroxysuccinimide may be adjusted to 50-100 mmol (1-2 eq.), and sphingosine or dihydrosphingosine may be used instead of phytosphingosine.

[0107] Workup: The solution was diluted with 80 mL of water, separated, and the organic phase was washed once with 80 mL of water and once with 80 mL of saturated salt. The aqueous phase was extracted once with 80 mL of dichloromethane, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the crude product was obtained by rotational evaporation under reduced pressure. The product was then purified by column chromatography to obtain the final product.

[0108] Deprotection reaction: The crude product (1 eq, 20 mmol) was dissolved in 20 mL of THF, and diethylamine (4 eq, 80 mmol) was slowly added dropwise. The reaction was allowed to proceed at room temperature until the crude product was confirmed to be completely reacted by TLC detection.

[0109] Workup: THF was removed by rotary evaporation, diluted with 60 mL of saturated saline solution, extracted with 80 mL of dichloromethane, the resulting organic phase was collected, dried over anhydrous sodium sulfate, filtered, and subjected to reduced-pressure rotary evaporation to obtain the crude product. The crude product was purified by column chromatography to obtain the final product.

[0110] Example 1 Alanine phytosphingosine ceramide was synthesized by method A and obtained as compound 1. JPEG2026513487000028.jpg26170 1 H NMR(400MHz,Methanol-d4)δ4.14(dt,J=6.5,4.5Hz,1H),3.79(dd,J=11.2,4.3Hz,1H),3.68(dd,J=11.3,6.5Hz,1H),3.58-3.43 (m,2H),2.93(t,J=6.5Hz,2H),2.48-2.32(m,2H),1.74-1.62(m,1H),1.57(s,1H),1.29(d,J=9.3Hz,24H),0.89(t,J=6.7Hz,3H)

[0111] Example 2 Threonine phytosphingosine ceramide was synthesized by method B and obtained as compound 2. JPEG2026513487000029.jpg26170 1 H NMR(400MHz,Methanol-d4)δ4.15(dt,J=5.9,4.4Hz,1H),4.01(qd,J=6.4,4.5Hz,1H),3.82-3.68(m,2H),3.57-3.48(m,2H),3.16(d, J=4.5Hz,1H),1.69(dd,J=12.3,9.3Hz,1H),1.54(d,J=10.9Hz,1H),1.34-1.29(s,24H),1.20(d,J=6.4Hz,3H),0.90(t,J=6.7Hz,3H)

[0112] Example 3 Proline phytosphingosine ceramide was synthesized by method A and obtained as compound 3. JPEG2026513487000030.jpg27170 1 H NMR(400MHz,Methanol-d4)δ4.09(q,J=5.1Hz,1H),3.80-3.70(m,2H),3.67(dd,J=8.7,5.3Hz,1H),3.58-3.44(m,2H),3 .02-2.87(m,2H),2.18-2.05(m,1H),1.86-1.62(m,4H),1.60-1.45(m,1H),1.30(d,J=8.6Hz,25H),0.90(t,J=6.7Hz,3H)

[0113] Example 4 Asparagine phytosphingosine ceramide was synthesized by method B and obtained as compound 4. JPEG2026513487000031.jpg26170 1H NMR(400MHz,Methanol-d4)δ4.17-4.09(m,1H),3.79(dd,J=11.3,4.4Hz,1H),3.76-3.64(m,2H),3.56-3.45(m,2H),2.66(dd,J=15. 3,5.1Hz,1H),2.49(dd,J=15.3,7.8Hz,1H),1.76-1.64(m,1H),1.54(d,J=11.3Hz,1H),1.30(d,J=8.5Hz,25H),0.90(t,J=6.7Hz,3H)

[0114] Example 5 Glutamine phytosphingosine ceramide was synthesized by method B and obtained as compound 5. JPEG2026513487000032.jpg27170 1 H NMR(400MHz,Chloroform-d)δ4.65(d,J=5.5Hz,1H),4.54(t,J=5.5Hz,1H),4.37(d,J=6.5Hz,1H),4.03(dd,J=8.5,4.3Hz,1H),3.98-3.84(m,1H), 3.52(dq,J=10.9,5.2Hz,1H),2.30-1.99(m,3H),1.86(ddd,J=12.3,9.0, 4.6Hz,1H),1.55-1.34(m,2H),1.23-1.18(s,24H),0.85(t,J=6.6Hz,3H)

[0115] Example 6 Leucine phytosphingosine ceramide was synthesized by method A and obtained as compound 6. JPEG2026513487000033.jpg26170 1 H NMR(400MHz,Methanol-d4)δ4.08(td,J=5.8,4.2Hz,1H),3.81-3.68(m,2H),3.56(t,J=5.9Hz,1H),3.51(ddd,J=8.7,5. 9,2.5Hz,1H),3.34(tt,J=6.3,3.1Hz,1H),1.75-1.49(m,4H),1.42-1.20(m,25H),0.98-0.91(m,6H),0.91-0.86(m,3H)

[0116] Example 7 Tryptophan phytosphingosine ceramide was synthesized by method A and obtained as compound 7. JPEG2026513487000034.jpg35170 1 H NMR(400MHz,Methanol-d4)δ7.62(d,J=7.9Hz,1H),7.34(d,J=8.0Hz,1H),7.12(s,1H),7.09(ddd,J=8 .2,6.9,1.2Hz,1H),7.04-6.97(m,1H),4.09(q,J=5.1Hz,1H),3.72-3.61(m,3H),3.44(ddd,J=9.0,6.3 ,2.5Hz,1H),3.39(dd,J=6.4,5.0Hz,1H),3.22(dd,J=14.2,5.6Hz,1H),2.95(dd,J=14.2,7.8Hz,1H), 1.63(ddt,J=12.4,10.2,2.4Hz,1H),1.51(q,J=7.7Hz,1H),1.29(d,J=10.1Hz,24H),0.96-0.83(m,3H)

[0117] Example 8 Serine phytosphingosine ceramide was synthesized by method B and obtained as compound 8. JPEG2026513487000035.jpg27170 1 H NMR(400MHz,Methanol-d4)δ4.17(q,J=4.8Hz,1H),3.85-3.66(m,4H),3.59-3.49(m,2H),3.44(t, J=5.5Hz,1H),1.77-1.63(m,1H),1.56(d,J=11.3Hz,1H),1.33-1.30(m,24H),0.92(t,J=6.7Hz,3H)

[0118] Example 9 Valinphytosphingosineceramide was synthesized by method A and obtained as compound 9. JPEG2026513487000036.jpg27170 1H NMR(400MHz,Methanol-d4)δ4.19-4.11(m,1H),3.81(dd,J=11.3,4.3Hz,1H),3.74(dd,J=11.3,5.9Hz,1H),3.61-3.50(m,2H),3.17(d,J=5.7H) z,1H),2.01(dq,J=13.4,6.8Hz,1H),1.73-1.50(m,2H),1.40-1.24(m,2 4H),1.00(d,J=6.8Hz,3H),0.96(d,J=6.9Hz,3H),0.92(t,J=6.7Hz,3H)

[0119] Example 10 Methionine phytosphingosine ceramide was synthesized by method B and obtained as compound 10. JPEG2026513487000037.jpg27170 1 H NMR(400MHz,Methanol-d4)δ4.15(td,J=5.8,4.2Hz,1H),3.82(dd,J=11.3,4.2 Hz,1H),3.74(dd,J=11.3,6.1Hz,1H),3.61-3.49(m,3H),2.58(dd,J=8.6,6.6H z,2H),2.12(s,3H),2.08-1.97(m,1H),1.86(dt,J=14.0,7.1Hz,1H),1.66(d,J =11.1Hz,1H),1.57(d,J=12.1Hz,1H),1.46-1.23(m,24H),0.92(t,J=6.8Hz,3H)

[0120] Example 11 Tyrosine phytosphingosine ceramide was synthesized by method B and obtained as compound 11. JPEG2026513487000038.jpg27170 1H NMR(400MHz,Methanol-d4)δ7.12(d,J=8.5Hz,2H),6.81-6.74(m,2H),4.17(dt,J=6.5 ,4.5Hz,1H),4.01(dd,J=8.3,6.2Hz,1H),3.83(dd,J=11.3,4.3Hz,1H),3.69(dd,J=11 .3,6.6Hz,1H),3.49-3.38(m,2H),3.12(dd,J=14.1,6.3Hz,1H),2.91(dd,J=14.1,8.3 Hz,1H),1.72-1.60(m,1H),1.54(d,J=10.5Hz,1H),1.28(s,24H),0.90(t,J=6.8Hz,3H)

[0121] Example 12 Histidine phytosphingosine ceramide was synthesized by method B and obtained as compound 12. JPEG2026513487000039.jpg33170 1 H NMR(400MHz,Methanol-d4)δ7.61(s,1H),6.92(s,1H),4.13(dt,J=6.4,4.6Hz,1H),3.79(dd,J=11.3,4.3Hz,1H),3.74-3.63(m,2H),3.52-3 .40(m,2H),3.02(dd,J=14.7,5.3Hz,1H),2.88(dd,J=14.6,7.4Hz,1H),1.75-1.63(m,1H),1.55(s,1H),1.28(s,24H),0.90(t,J=6.7Hz,3H)

[0122] Example 13 L-aspartate-4-tert-butyl ester phytosphingosine ceramide was synthesized by method C and obtained as compound 13. JPEG2026513487000040.jpg26170 1H NMR(400MHz,Methanol-d4)δ4.10(q,J=5.0Hz,1H),3.75(qd,J=11.3,5.1Hz,2H),3.63(dd,J=7.2,5.4Hz,1H),3.59-3.47(m,2H),2.70(dd,J =16.5,5.4Hz,1H),2.52(dd,J=16.5,7.2Hz,1H),1.73-1.60(m,1H),1.54(t,J=8.0Hz,1H),1.46(s,9H),1.28(s,24H),0.90(t,J=6.7Hz,3H)

[0123] Example 14 L-1-tert-butyl aspartate phytosphingosine ceramide was synthesized by method C and obtained as compound 14. JPEG2026513487000041.jpg27170 1 H NMR(400MHz,Methanol-d4)δ4.14(dt,J=6.3,4.4Hz,1H),3.79(ddd,J=9.4,4.3,1.7Hz,2H),3.68(dd,J=11.5,6.3Hz,1H),3.57-3.46(m,2H),2.71( dd,J=15.4,4.3Hz,1H),2.60(dd,J=15.5,7.9Hz,1H),1.71-1.67(m,1H), 1.59-1.51(m,1H),1.48(s,9H),1.36-1.22(m,24H),0.90(t,J=6.7Hz,3H)

[0124] Example 15 Alanine sphingosine ceramide was synthesized by method A and obtained as compound 15. JPEG2026513487000042.jpg26170 1H NMR(400MHz,Methanol-d4)δ5.70-5.61(m,1H),5.52-5. 40(m,1H),4.06(t,J=7.4Hz,1H),3.89-3.82(m,1H),3.68(d,J=5.0Hz,2H),2.93(t,J=6.5Hz, 2H),2.48-2.32(m,2H),2.15(t,J=7.6Hz,2H),1.76-1.64(m,1H),1.59-1.55(m,1H),1.28-1. 22(m,20H),0.91(t,J=6.8Hz,3H)

[0125] Example 16 Alanine dihydrosphingosine ceramide was synthesized by method A and obtained as compound 16. JPEG2026513487000043.jpg26170 1 H NMR(400MHz,Methanol-d4)δ4.12(td,J=6.1,4.1Hz,1H),3.91-3.81(m,2H),3.79-3.74(m,1H),2.9 1(t,J=6.5Hz,2H),2.51-2.34(m,2H),1.47-1.39(m,3H),1.28-1.22(m,25H),0.88(t,J=6.8Hz,3H)

[0126] Example 17 Synthesis of glycylglycine-phytosphingosine ceramide Step 1: Dissolve Boc-protected glycylglycine (1.0 eq, 50 mmol) and N-hydroxysuccinimide (1.5 eq, 75 mmol) in 100 mL of dichloromethane, add DCC (1.5 eq, 75 mmol) at room temperature, and react overnight at room temperature. Then add phytosphingosine (1 eq, 50 mmol) and react with stirring until TLC detection confirms that phytosphingosine has reacted completely.

[0127] Workup: Dilute with 80 mL of DCM, filter to remove solids, wash the filtrate once each with 80 mL of water and 80 mL of saturated saline, extract the collected aqueous phase once with 80 mL of dichloromethane, combine with the organic phase, dry over anhydrous sodium sulfate, filter, and obtain the crude product by rotational evaporation under reduced pressure. The product was then purified by column chromatography to obtain the final product.

[0128] Step 2: Dissolve the product obtained in the previous step (1 eq, 30 mmol) in 30 mL of DCM, add 15 mL of TFA, and allow to react overnight at room temperature with stirring until the starting materials have reacted completely, as confirmed by TLC detection. Workup: After removing the reaction mixture by rotary evaporation, the pH of the reaction system was adjusted to approximately 10 by adding saturated sodium carbonate solution. 30 mL of DCM was added, followed by liquid-liquid separation. The organic phase was washed twice with 30 mL of water, and the combined aqueous phase was extracted once with 20 mL of DCM. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and subjected to reduced-pressure rotary evaporation to obtain the crude product. The crude product was recrystallized with methanol, filtered, and purified to obtain a white solid product. 1 H NMR(400MHz,Methanol-d4)δ4.15(dt,J=6.3,4.7Hz,1H),3.91(d,J=1.3Hz,2H),3.78(dd,J=11.2,4.4Hz,1H),3.73-3.62(m,1H),3 .57-3.45(m,2H),3.33(s,2H),1.67(td,J=9.5,8.7,5.1Hz,1H),1.52(d,J=14.9Hz,1H),1.30-1.26(m,24H),0.89(t,J=6.7Hz,3H)

[0129] Example 18 Synthesis of dipeptide-1-phytosphingosine (snake venom peptide-phytosphingosine)ceramide Step 1: Dissolve Boc-protected dipeptide-1 (1.0 eq, 30 mmol), EDCI (1.5 eq, 45 mmol), and N-hydroxysuccinimide (1.5 eq, 45 mmol) in 60 mL of dichloromethane and react at room temperature for 30 minutes. Then add phytosphingosine (1 eq, 45 mmol) and react with stirring at room temperature until TLC detection confirms that phytosphingosine has reacted completely.

[0130] Workup: Dilution with 50 mL of water, liquid-liquid separation, further washing of the organic phase with 80 mL of water and 80 mL of saturated saline, respectively, extraction of the aqueous phase once with 60 mL of dichloromethane, combination with the organic phase, drying over anhydrous sodium sulfate, filtration, and rotational evaporation under reduced pressure to obtain the crude product, which was then purified by column chromatography to obtain the final product.

[0131] Step 2: Dissolve the product obtained in the previous step (1 eq, 10 mmol) in 10 mL of DCM, add 5 mL of TFA, and allow to react overnight at room temperature with stirring until the starting materials have reacted completely, as confirmed by TLC detection.

[0132] Workup: The reaction mixture was removed by rotary evaporation, saturated sodium carbonate solution was added to adjust the pH of the reaction system to approximately 10, 30 mL of DCM was added, and the mixture was separated. The organic phase was washed twice with 30 mL of water, the combined aqueous phase was extracted once with 20 mL of DCM, the organic phase was combined, dried over anhydrous sodium sulfate, filtered, and then subjected to reduced-pressure rotary evaporation to obtain the crude product. The crude product was purified by column chromatography to obtain the dipeptide-1-phytosphingosineceramide product. 1H NMR(400MHz,Methanol-d4)δ4.44(dd,J=8.5,3.9Hz,1H),4.10(td,J=5.9,4.1Hz,1H),3.94-3.42(m,6H),3.11-3.08(m,2H),2.70( dt,J=8.7,6.0Hz,2H),2.24(td,J=8.2,4.9Hz,1H),2.15-1.88(m,3H),1.73-1.50(m,2H),1.32-1.29(s,24H),0.92(t,J=6.7Hz,3H)

[0133] Example 19 Synthesis of dipeptide-2-phytosphingosineceramide JPEG2026513487000046.jpg22170 was synthesized using the method for synthesizing dipeptide-1-phytosphingosineceramide. 1 H NMR(400MHz,Chloroform-d)δ7.07(d,J=8.3Hz,2H),6.69(d,J=8.3Hz,2H),4.55(dd ,J=8.7,5.9Hz,1H),4.12-4.03(m,1H),3.70(dtd,J=21.7,10.7,10.2,3.6Hz,2H),3. 49(ddd,J=22.8,9.6,5.7Hz,3H),3.03(td,J=14.6,14.2,6.4Hz,1H),2.82(dd,J=13. 7,9.0Hz,1H),1.67-1.49(m,2H),1.28-1.24(m,J=9.8Hz,27H),0.89(t,J=6.7Hz,3H)

[0134] Example 20 Synthesis of hexapeptide-1-sphingosineceramide Step 1: Synthesis of sphingosine and succinic anhydride JPEG2026513487000047.jpg351703 In a 1L neck flask, succinic anhydride (3.78g, 37.8 mmol) and sphingosine (9.5g, 31.5 mmol) were added and dissolved in 100 mL of THF solution. DIPEA (8.2g, 63 mmol) was added, and the mixture was allowed to react with stirring at room temperature until complete reaction was confirmed by TLC detection. The mixture was extracted twice with dichloromethane and hydrochloric acid aqueous solution, and once with dichloromethane and saturated saline solution. The organic phases were combined and purified by column chromatography to obtain white solid product I (8.1 g).

[0135] Step 2: Sequential synthesis of hexapeptide-1 1. Synthesis of MBHA Linker 7 mmol (10 g, sample injection volume: 0.69 mmol / g) of MBHA resin was weighed, swollen in 100 mL of DMF for 2 hours, filtered by suction, and dried. It was washed three times with 100 mL of dry DMF and filtered by suction until dry. 7.5 g of Fmoc-Linker and 2.2 g of HOBT were weighed, dissolved in 100 mL of DMF, 2.6 mL of DIC was added under ice bath, and the mixture was activated in the solution for approximately 5 minutes. The mixture was then introduced into a reaction column and reacted at room temperature for 1 hour. The progress of the reaction was confirmed by sampling, the resin was washed three times with DMF (100 mL / 1 time / 3 min), and filtered by suction until dry.

[0136] 2. Synthesis of Fmoc-Trp(Boc)-OH The sample was deprotected twice with a 20% piperidine / DMF solution (100 mL / time), and after 10 min + 10 min deprotection, it was washed six times with DMF (100 mL / time / 3 min), and filtered by aspirate until dry. The ninhydrin test showed that K + The following was demonstrated. 8.9 g of Fmoc-Trp(Boc)-OH and 2.2 g of HOBT were weighed, dissolved in 100 mL of DMF, 2.6 mL of DIC was added under ice bath, and the mixture was activated in the solution for approximately 5 minutes. The mixture was then introduced into a reaction column and reacted at room temperature for 1 hour. The progress of the reaction was confirmed by sampling, the resin was washed three times with DMF (100 mL / 1 time / 3 min), and the mixture was filtered by suction until dry.

[0137] 3. Synthesis of Fmoc-Arg(Pbf)-OH A 20% piperidine / DMF solution was added, and the sample was deprotected twice (100 mL / time). After deprotection for 10 min + 10 min, the sample was washed six times with DMF (100 mL / time / 3 min) and filtered by aspirate until dry. The ninhydrin test showed that K + The following was demonstrated. 9.1 g of Fmoc-Arg(Pbf)-OH and 2.2 g of HOBT were weighed, dissolved in 100 mL of DMF, 2.6 mL of DIC was added under ice bath, and the mixture was activated in the solution for approximately 5 minutes. The mixture was then introduced into a reaction column and reacted at room temperature for 1 hour. The progress of the reaction was confirmed by sampling, the resin was washed three times with DMF (100 mL / 1 time / 3 min), and filtered by suction until dry.

[0138] 4. Synthesis of Fmoc-D-Phe-OH A 20% piperidine / DMF solution was added, and the mixture was deprotected twice (100 mL / time). After deprotection for 10 min + 10 min, the mixture was washed six times with DMF (100 mL / time / 3 min), filtered by suction until dry, and the progress of the reaction was confirmed by sampling. 8.1 g of Fmoc-D-Phe-OH and 2.2 g of HOBT were added, dissolved in 100 mL of DMF, and 2.6 mL of DIC was added under ice bath. The mixture was activated in the solution for approximately 5 min, introduced into a reaction column, reacted at room temperature for 1 hour, and the progress of the reaction was confirmed by sampling. The resin was washed three times with DMF (100 mL / time / 3 min), and filtered by suction until dry.

[0139] 5. Synthesis of Fmoc-His(Trt)-OH The solution was deprotected twice by adding a 20% piperidine / DMF solution (100 mL / time), and after deprotection for 10 min + 10 min, the mixture was washed six times with DMF (100 mL / time / 3 min), filtered by suction until dry, and the progress of the reaction was confirmed by sampling. 12.8 g of Fmoc-His(Trt)-OH and 2.2 g of HOBT were weighed, dissolved in 100 mL of DMF, activated in solution for approximately 5 min by adding 2.6 mL of DIC under an ice bath, introduced into a reaction column, reacted at room temperature for 1 hour, the progress of the reaction was confirmed by sampling, the resin was washed three times with DMF (100 mL / time / 3 min), and filtered by suction until dry.

[0140] 6. Synthesis of Fmoc-Ala-OH The reaction was performed twice by adding a 20% piperidine / DMF solution (100 mL / time), and after deprotection for 10 min + 10 min, the mixture was washed six times with DMF (100 mL / time / 3 min), filtered by suction until dry, and the progress of the reaction was confirmed by sampling. 5.2 g of Fmoc-Ala-OH and 2.3 g of HOBT were weighed, dissolved in 100 mL of DMF, activated in solution for approximately 5 min by adding 2.6 mL of DIC under an ice bath, introduced into a reaction column, reacted at room temperature for 1 hour, the progress of the reaction was confirmed by sampling, the resin was washed three times with DMF (100 mL / time / 3 min), and filtered by suction until dry.

[0141] 7. Synthesis of Fmoc-Nle-OH The reaction was performed twice by adding a 20% piperidine / DMF solution (100 mL / time), and after deprotection for 10 min + 10 min, the mixture was washed six times with DMF (100 mL / time / 3 min), filtered by suction until dry, and the progress of the reaction was confirmed by sampling. 7.4 g of Fmoc-Nle-OH and 2.3 g of HOBT were weighed, dissolved in 100 mL of DMF, activated in solution for approximately 5 min by adding 2.6 mL of DIC under an ice bath, introduced into a reaction column, reacted at room temperature for 1 hour, the progress of the reaction was confirmed by sampling, the resin was washed three times with DMF (100 mL / time / 3 min), and filtered by suction until dry.

[0142] Step 3: Synthesis of hexapeptide-1-sphingosineceramide by sequential synthesis of compound I and hexapeptide-1. JPEG2026513487000048.jpg32170 Add a 20% piperidine / DMF solution and deprotect twice (100 mL / time), deprotect for 10 min + 10 min, wash six times with DMF (100 mL / time / 3 min), filter by suction until dry, and confirm the progress of the reaction by sampling. Weigh 5.8 g of compound I synthesized in step 1 and 2.3 g of HOBt, dissolve in 100 mL of DMF, add 2.6 mL of DIC under an ice bath, activate in solution for about 5 min, introduce into a reaction column, react at room temperature for 1 hour, confirm the progress of the reaction by sampling, wash the resin three times with DMF (100 mL / time / 3 min), and filter by suction until dry. Wash the resin three times with methanol (100 mL / time / 3 min), filter by suction until dry to obtain crude peptide resin, and dry in a vacuum drying oven. The product obtained by the sequential synthesis of compound I and hexapeptide-1 was named hexapeptide-1-399.

[0143] Decomposition and preparative separation of hexapeptide-1-399: The weight of the dried resin obtained by vacuum drying was approximately 17.8 g. A 200 mL decomposition solution (TFA:EDT:TIPS:H2O = 92.5:2.5:2.5:2.5) was prepared and decomposed at room temperature for 2 hours. 2 L of frozen isopropyl ether was added to precipitate, and a white solid was released. The white solid was filtered by suction and dried, then dissolved in water and acetonitrile. Separation was performed using a half-separation apparatus under a 30-70 gradient (mobile phase: 1‰ trifluoroacetic acid aqueous solution and acetonitrile). The resulting pure fraction was concentrated under reduced pressure and freeze-dried. The mass spectrometry results of the obtained compound are shown in Figure 16.

[0144] Example 21 Synthesis of hexapeptide-1-phytosphingosineceramide Step 1: Synthesis of phytosphingosine and succinic anhydride JPEG2026513487000049.jpg361703 In a 1L neck flask, succinic anhydride (3.78g, 37.8 mmol) and phytosphingosine (10g, 31.5 mmol) were added and dissolved in 100 mL of THF solution. DIPEA (8.2g, 63 mmol) was added, and the mixture was stirred at room temperature until complete reaction was confirmed by TLC detection. The mixture was extracted twice with dichloromethane and hydrochloric acid aqueous solution, and once with dichloromethane and saturated saline solution. The organic phases were combined and purified by column chromatography to obtain white solid product I (8.2g).

[0145] Step 2: Synthesize hexapeptide-1-phytosphingosinecerami by sequential synthesis of compound II and hexapeptide-1. The reaction was carried out according to Step 3 of Example 20, and the mass spectrometry results of the obtained compound are shown in Figure 17.

[0146] Example 22 Synthesis of hexapeptide-8-phytosphingosineceramide The sequential synthesis of hexapeptide-8 was carried out according to the synthesis method for hexapeptide-1. The reaction was carried out according to Example 21, and the results of the mass spectrometry analysis of the obtained compound are shown in Figure 18.

[0147] Example 23 Synthesis of hexapeptide-9-sphingosineceramide The sequential synthesis of hexapeptide-9 was carried out according to the synthesis method of hexapeptide-1. The reaction was carried out according to Example 20, and the results of the mass spectrometry analysis of the obtained compound are shown in Figure 19.

[0148] Example 24 Synthesis of hexapeptide-9-phytosphingosineceramide The reaction was carried out according to Example 21, and the results of the mass spectrometry analysis of the obtained compound are shown in Figure 20.

[0149] Example 25 Synthesis of tetrapeptide-5-sphingosineceramide The sequential synthesis of tetrapeptide-5 was carried out according to the synthesis method for hexapeptide-1. The reaction was carried out according to Example 20, and the results of the mass spectrometry analysis of the obtained compound are shown in Figure 21.

[0150] Example 26 Synthesis of tetrapeptide-5-phytosphingosineceramide The reaction was carried out according to Example 21, and the results of the mass spectrometry analysis of the obtained compound are shown in Figure 22.

[0151] Example 27 Synthesis of Octapeptide-3-PhytosphingosineCeramide The sequential synthesis of octapeptide-3 was carried out according to the synthesis method for hexapeptide-1. The reaction was carried out according to Example 21, and the results of the mass spectrometry analysis of the obtained compound are shown in Figure 23.

[0152] Example 28 Synthesis of nonapeptide-1-sphingosineceramide The sequential synthesis of nonapeptide-1 was carried out according to the synthesis method for hexapeptide-1. The reaction was carried out according to Example 20, and the mass spectrometry results of the obtained compound are shown in Figure 24.

[0153] Example 29 Synthesis of nonapeptide-1-phytosphingosineceramide The reaction was carried out according to Example 21, and the results of the mass spectrometry analysis of the obtained compound are shown in Figure 25.

[0154] Example 30 Antioxidant experiment: Nitrogen radical scavenger DPPH and oxygen radical scavenger PTIO were used. Measurement of DPPH radical removal rate Samples of corresponding concentrations were mixed with 0.1 mol / L DPPH and anhydrous ethanol solutions in a 1:1 volume ratio, while the blank group consisted of an equivolume mixture of DPPH and anhydrous ethanol in a 1:1 ratio. After reacting at room temperature in the dark for 30 minutes, the absorbance at 517 nm was measured. If the absorbance of the sample reaction solution with a different concentration than DPPH is denoted as A1, the absorbance of the sample reaction solution with a different concentration than anhydrous ethanol is denoted as A2, and the absorbance of the DPPH and anhydrous ethanol reaction solution is denoted as A3, then the DPPH removal rate of the sample = [1 - (A1 - A2) / A3] × 100%.

[0155] Measurement of PTIO radical removal rate Samples of corresponding concentrations were mixed with 0.6 mmol / L PTIO and anhydrous ethanol solution in a 1:2 volume ratio, while the blank group consisted of an equivolume mixture of PTIO and anhydrous ethanol in a 1:2 ratio. After reacting at room temperature in the dark for 30 minutes, the absorbance at 557 nm was measured. If the absorbance of the sample reaction solution with a different concentration than PTIO is denoted as A1, the absorbance of the sample reaction solution with a different concentration than anhydrous ethanol is denoted as A2, and the absorbance of the PTIO and anhydrous ethanol reaction solution is denoted as A3, then the PTIO removal rate of the sample = [1 - (A1 - A2) / A3] × 100%.

[0156] Figures 1-12 are histograms of the antioxidant experiment results for compounds 1, 2, 3, 4, 6, 7, 8, 9, 10, 11, 13, and 14, respectively. The amino acid ceramide compounds of the present invention have good antioxidant performance and higher scavenging performance against oxygen radicals than against nitrogen radicals.

[0157] Example 31 Anti-glycation experiment Under sterile conditions, 4 mL each of sterilized bovine serum albumin solution and glucose solution, lined with sterile film, were added to a sterile flask and mixed uniformly. Then, 4 mL each of sample solution and phosphate buffer of different concentrations were added, and the mixture was incubated at 37°C in the dark for 10 days. Aminoguanidine was used instead of the sample as the positive control group, and phosphate was used instead of the sample as the blank control group. AGEs content was measured on day 10 of the reaction. Three parallel measurements were performed for each group.

[0158] Three mL of glycosylated material was collected, and the fluorescence value Fs of the final glycosylated product was measured at an excitation wavelength of 340 nm and an emission wavelength of 420 nm, representing the total fluorescence AGEs content. The value measured in a phosphoric acid solution is F0. Suppression rate %=(F0-Fs) / F0×100 Figure 13 shows histograms of the anti-glycation experiment results for compounds 1, 2, 3, and 4. Figure 14 shows histograms of the anti-glycation experiment results for compounds 6, 7, 8, and 9. Figure 15 shows histograms of the anti-glycation experiment results for compounds 10, 11, 13, and 14. All of the amino acid ceramide compounds of the present invention have good anti-glycation effects.

[0159] Example 32 Moisturizing function test Equipment: Constant temperature and humidity waterproof incubator, thermometer and hygrometer, sealed dryer, volumetric bottles, precision electronic balance (0.0001 g increments), dryer. Reagents: Hyaluronic acid (HA), compounds synthesized in Examples 1-13, dried fine silica gel, potassium carbonate, ammonium sulfate. Solution preparation: Saturated potassium carbonate solution (43% humidity), saturated ammonium sulfate solution (81% humidity). Moisture absorption rate: Labels were attached to the weighing bottles, samples were placed inside, the bottom of the bottles was lined with a lining, and the weighing bottles were placed in a vacuum drying box and dried for 1 hour. The weighing bottles were removed and each was weighed on an electronic balance, and the initial weight at 0h was recorded. The weighing bottles and saturated ammonium sulfate solution were placed in the drying box and sealed, and placed in a waterproof incubator (20-25°C). The weighing bottles were removed and weighed at 4, 8, 12, and 24h, and the data was recorded. Hyaluronic acid (HA) solution was measured in the same manner as a control sample. The above steps were repeated using saturated potassium carbonate solution instead of saturated ammonium sulfate solution. Calculation: Moisture absorption rate (%) = (M-M0) ÷ M0 × 100% M: Sample weight after moisture absorption M0: Sample weight after vacuum drying and when the weight becomes constant

[0160] Figure 26 shows the moisture absorption rate-time curves for hyaluronic acid, glycylglycine-phytosphingosine, snake venom peptide-phytosphingosine, and dipeptide-2-phytosphingosine (from left to right, top to bottom, and so on) at an ambient humidity of 81%. Figure 27 shows the moisture absorption rate-time curves for tetrapeptide-5-phytosphingosine, tetrapeptide-5-sphingosine, hexapeptide-1-phytosphingosine, and hexapeptide-1-sphingosine at an ambient humidity of 81%. Figure 28 shows the moisture absorption rate-time curves for hexapeptide-8-phytosphingosine, hexapeptide-9-phytosphingosine, hexapeptide-9-sphingosine, and octapeptide-3-phytosphingosine at an ambient humidity of 81%. Figure 29 shows the moisture absorption rate-time curves for nonapeptide-1-phytosphingosine, nonapeptide-1-sphingosine, and hyaluronic acid and glycylglycine-phytosphingosine at 81% ambient humidity and 43% ambient humidity. Figure 30 shows the moisture absorption rate-time curves for snake venom peptide-phytosphingosine, dipeptide-2-phytosphingosine, tetrapeptide-5-phytosphingosine, and tetrapeptide-5-sphingosine at 43% ambient humidity. Figure 31 shows the moisture absorption rate-time curves for hexapeptide-1-phytosphingosine, hexapeptide-1-sphingosine, hexapeptide-8-phytosphingosine, and hexapeptide-9-phytosphingosine at 43% ambient humidity. Figure 32 shows the moisture absorption rate-time curves for hexapeptide-9-sphingosine, octapeptide-3-phytosphingosine, nonapeptide-1-phytosphingosine, and nonapeptide-1-sphingosine at an ambient humidity of 43%. The ceramide compounds of the present invention exhibit good moisture absorption effects under two humidity conditions.

[0161] The above are merely specific embodiments of the present invention, and the scope of protection of the present invention is not limited thereto. Modifications or substitutions that a person skilled in the art can easily conceive within the technical scope disclosed herein should be included within the scope of protection of the present invention. Therefore, the scope of protection of the present invention is determined by the content described in the claims.

Claims

1. A ceramide derived from an amino acid, having a structure of general formula I, or an isomer thereof. And the R １ These are residues formed by the condensation of polypeptides, The aforementioned R ３ These are selected from residues formed by the condensation of alanine, threonine, proline, asparagine, glutamine, leucine, tryptophan, serine, valine, methionine, tyrosine, histidine, L-aspartate-4-tert-butyl ester, L-aspartate-1-tert-butyl ester, glycine, isoleucine, phenylalanine, lysine, arginine, cysteine, L-glutamate-5-tert-butyl ester, L-glutamate-1-tert-butyl ester, and polypeptides. The polypeptide is formed by the condensation of 2 to 10 amino acids. R ２ is selected from -C １５ H ２９ , -C １５ H ３１ , -C １５ H ２７ , -CHOHC １４ H ２７ , -CHOHC １４ H ２９ .)

2. The aforementioned R ３ The ceramide is derived from an amino acid according to claim 1, selected from residues obtained by the condensation of alanine, threonine, proline, asparagine, glutamine, leucine, tryptophan, serine, valine, methionine, tyrosine, histidine, L-aspartate-4-tert-butyl ester, L-aspartate-1-tert-butyl ester, glycine, isoleucine, phenylalanine, lysine, arginine, cysteine, L-glutamate-5-tert-butyl ester, L-glutamate-1-tert-butyl ester, dipeptide, tetrapeptide, hexapeptide, octapeptide, and nonapeptide.

3. The aforementioned R ３ The ceramide is derived from an amino acid according to claim 2, selected from residues obtained by the condensation of alanine, threonine, proline, asparagine, glutamine, leucine, tryptophan, serine, valine, methionine, tyrosine, histidine, L-aspartate-4-tert-butyl ester, L-aspartate-1-tert-butyl ester, glycylglycine, snake venom peptide intermediate, alanyl-L-tyrosine, tetrapeptide-5, hexapeptide-1, hexapeptide-8, hexapeptide-9, octapeptide-3, and nonapeptide-1.

4. The aforementioned R １ The ceramide is derived from the amino acid described in claim 1, wherein the residue is a condensed form of a dipeptide, tetrapeptide, hexapeptide, octapeptide, or nonapeptide.

5. The aforementioned R １ The ceramide is derived from the amino acid according to claim 4, wherein the residue is a condensed residue of glycylglycine, snake venom peptide intermediate, alanyl-L-tyrosine, tetrapeptide-5, hexapeptide-1, hexapeptide-8, hexapeptide-9, octapeptide-3, and nonapeptide-1.

6. The aforementioned R ２ The structure is as follows: A ceramide derived from the amino acid described in claim 1, which is one selected from the above.

7. The aforementioned R ２ The structure is as follows: A ceramide derived from the amino acid described in claim 6, which is one selected from the above.

8. The following compounds: A ceramide derived from the amino acid described in claim 7, which is one selected from the above.

9. A method for synthesizing ceramide derived from an amino acid according to any one of claims 1 to 8, M is R ３ If that is the case, Step S1: Polypeptides or amino acids, sphingoid bases protected with Boc or Fmoc The steps include: reacting a condensing agent and a coupling agent to obtain compound C, Step S2: A step of removing the protecting group Boc or Fmoc from compound C to obtain the product, Includes, M

10. M is R ３ In this case, the condensing agent is EDCI or DCC, the coupling agent is N-hydroxysuccinimide, the molar ratio of the Boc or Fmoc-protected amino acid or polypeptide, sphingoid base, condensing agent, and coupling agent is (1-1.5):1:(1-2):(1-2), and the solvent for the S1 reaction is dichloromethane. M In this case, the condensing agent is HOBt and DIC, and the polypeptide and succinic acid are bonded to a sphingoid base. The synthesis method according to claim 9, wherein the molar ratio of HOBt and DIC is (1-1.5):1:(1-2):(1-2), and the solvent for the reaction is DMF.

11. Use of ceramide derived from an amino acid according to any one of claims 1 to 8 in cosmetics, health foods, or pharmaceuticals.

12. The use according to claim 11, wherein the cosmetic is a cosmetic essence oil or a cosmetic in an anhydrous form.

13. The use according to claim 11, wherein the ceramide derived from the amino acid has at least one of the functions of moisturizing, skin barrier repair, tissue healing, antioxidant, and anti-glycation.

14. A composition comprising, as an active ingredient, ceramide derived from an amino acid according to any one of claims 1 to 8, an isomer thereof, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof.

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