Ceramide compound containing conjugated carboxylic acid as well as preparation method and application of ceramide compound

Abstract

The invention belongs to the technical field of biological medicines, and relates to a ceramide compound shown in a chemical formula I: # imgabs0 #, in which R1 is selected from residues after condensation of ferulic acid, sinapic acid, abscisic acid, isotretinoin, tretinoin, fusidic acid, royal jelly acid, sorbic acid, caffeic acid or trans-cinnamic acid, and R2 is selected from one of the following structures:-C15H29,-C15H31,-C15H27,-CHOHC14H27 and-CHOHC14H29. The invention also discloses a preparation method and application of the ceramide compound. Effective carboxylic acid reacts with sphingosine alkali to obtain a ceramide compound with a novel structure, and molecular fragments, with physiological activity, of the effective carboxylic acid are introduced into ceramide, so that the original efficacy of molecules can be improved, and the physicochemical properties of the ceramide compound can be improved, for example, the physical and chemical properties of the ceramide compound can be improved. The oxidation resistance of the ceramide compound is improved.

Description

[0001] This application is a divisional application with application number 202210939967.0, application date August 5, 2022, and invention name "Ceramide compounds containing conjugated carboxylic acids, preparation methods and applications thereof". Technical Field

[0002] The present invention belongs to the field of biomedicine technology, and specifically relates to ceramide compounds containing conjugated carboxylic acids, and preparation methods and applications thereof. Background Art

[0003] Ceramide is one of the main components of the intercellular lipids that make up the stratum corneum of the skin, and is used to prevent excessive water loss caused by evaporation and maintain the structure of the stratum corneum. The stratum corneum acts as a barrier to protect the underlying tissue from harmful substances or microorganisms from the external environment. When differentiated keratinocytes fall off the stratum corneum, the intercellular lipids of the keratin form a thin sheet structure, which helps to maintain the basic functions of the skin. Intercellular lipids are composed of components such as ceramide, cholesterol, and free fatty acids, and among these components, ceramide is known to play an important role in the water retention and barrier function of the stratum corneum. It is known that a decrease in the ceramide content in the stratum corneum increases water evaporation and worsens various skin diseases. In addition, it is known that skin with a reduced ceramide content in the stratum corneum caused by skin aging or external stimulation can be restored to a normal state by external supplementation of ceramide. Therefore, various natural animal and plant products containing ceramide have been studied. The amount of these ceramides present in animals, plants, and microorganisms is extremely small, so there are problems such as difficulty in extraction and high production costs. In addition, naturally derived ceramides have very low solubility in substances such as solvents commonly used in cosmetics, so it is difficult to make products contain an amount that can fully exert their effectiveness.

[0004] Due to the widespread demand for functional ceramides in the market, it is necessary to research and develop ceramide derivatives that can mimic natural ceramides and have enhanced efficacy, which can solve the shortcomings of natural ceramides and can be produced commercially. Summary of the invention

[0005] The purpose of the present invention is to provide a class of novel ceramide compounds, which contain conjugated carboxylic acids. Conjugated carboxylic acids refer to carboxylic acids in which a carboxyl group is conjugated with a carbon-carbon double bond. The carbon-carbon double bond refers to a straight-chain -CH=CH-, and does not include cyclization such as cyclohexene, nor does it include carbon-carbon bonds of aromatic rings and heterocycles.

[0006] Another object of the present invention is to provide a method for synthesizing ceramide compounds.

[0007] Another object of the present invention is to provide uses of ceramide compounds.

[0008] In order to achieve one of the above purposes, the present invention adopts the following technical solution:

[0009] In the first aspect of the present invention, a ceramide compound has a structure of the general formula I or an enantiomer or diastereomer of the general formula I:

[0010]

[0011] Among them, R 1 A residue selected from the condensation of ferulic acid, sinapic acid, abscisic acid, isotretinoin, retinoic acid, fusidic acid, royal jelly acid, sorbic acid, caffeic acid or trans-cinnamic acid,

[0012] R 2 Select one of the following structures:

[0013] -C 15 H 29 , -C 15 H 31 , -C 15 H 27 , -CHOHC 14 H 27 , -CHOHC 14 H 29 .

[0014] The residue after condensation refers to the residual carboxylic acid fragment R after the carboxyl group of the corresponding carboxylic acid RCOOH condenses with the amino group of the sphingoid base to form a peptide bond. For example, the residue after condensation of royal jelly acid is The residue after condensation of ferulic acid is

[0015] Among them, ferulic acid, as a phenolic plant ingredient, has strong antioxidant activity and greatly promotes human health; ferulic acid can significantly increase the content of glutathione and nicotinamide adenine dinucleotide phosphate in irradiated cells, and has a protective effect on irradiated endothelial cells.

[0016] Isotretinoin is suitable for severe acne, especially nodular cystic acne. It can also be used for diseases such as pityriasis rubra pilaris.

[0017] Retinoic acid mainly affects bone growth and promotes epithelial cell proliferation, differentiation, keratin dissolution and other metabolic effects; it is used to treat diseases such as acne vulgaris, psoriasis, ichthyosis, lichen planus, pityriasis rubra pilaris, follicular keratosis, squamous cell carcinoma and melanoma.

[0018] Sorbic acid is a food additive and the most widely used preservative in the world. It has high antibacterial properties and can inhibit the growth and reproduction of mold.

[0019] Royal jelly acid, also known as 10-hydroxy-2-decenoic acid, is a component of royal jelly and is an unsaturated fatty acid. Royal jelly acid has the functions of enhancing the body's resistance, anti-bacterial and anti-inflammatory, and strongly inhibiting various cells such as lymphoma and breast cancer. It also has the functions of strengthening the body, preventing hair loss, and treating acute radiation damage and chemical damage.

[0020] Furthermore, the R 2 Select one of the following structures:

[0021]

[0022] Furthermore, the R 2 Select one of the following structures:

[0023]

[0024] They correspond to sphingosine, dihydrosphingosine, and phytosphingosine respectively.

[0025] Furthermore, the R 1 The residue is selected from the condensation residues of ferulic acid, trans-cinnamic acid, royal jelly acid, retinoic acid, sorbic acid, caffeic acid or sinapic acid.

[0026] Furthermore, the R 1 The residue is selected from the condensation residues of ferulic acid, royal jelly acid, retinoic acid, sorbic acid, caffeic acid or sinapic acid.

[0027] Furthermore, the R 1 Selected from the residues after condensation of ferulic acid.

[0028] Furthermore, the R 1 Selected from the residues after condensation of trans-cinnamic acid.

[0029] Furthermore, the R 1 Selected from the residues obtained by condensation of royal jelly acid.

[0030] Furthermore, the R 1 Selected from the residues after condensation of retinoic acid.

[0031] Furthermore, the R 1 Selected from the residues after condensation of sorbic acid.

[0032] Furthermore, the R 1 is selected from the residues after condensation of caffeic acid. 1 The ceramide compound is selected from the residue after condensation of sinapinic acid.

[0033]

[0034]

[0035] A second aspect of the present invention is a method for preparing a ceramide compound, comprising the following steps:

[0036]

[0037] Compound S1 reacts with N-hydroxysuccinimide and a condensation agent to obtain compound S2;

[0038] Compound S2 reacts with a sphingoid base to give compound I.

[0039] Furthermore, the molar ratio of the compound S1, N-hydroxysuccinimide, condensing agent and sphingoid base is 1:(1-2):(1-2):(0.8-1).

[0040] Furthermore, the condensation agent is DCC.

[0041] Furthermore, the solvent of the reaction is THF.

[0042] A method for preparing a ceramide compound comprises the following steps:

[0043]

[0044] Compound S1 reacts with a sphingoid base and a condensing agent to obtain compound I.

[0045] Furthermore, the condensing agent includes EDCI and HOBT.

[0046] Furthermore, the molar ratio of the compound S1, EDCI, HOBT and sphingoid base is 1:(1-2):(1-2):(0.8-1).

[0047] Furthermore, the solvent of the reaction is DCM.

[0048] The third aspect of the present invention is the use of ceramide compounds as antioxidants, especially in cosmetics, health products and medicines.

[0049] The present invention has the following beneficial effects:

[0050] The present invention reacts functional carboxylic acid with sphingosine base to obtain a class of ceramide compounds with novel structures. The introduction of molecular fragments of functional carboxylic acid with physiological activity into ceramide will enhance the original efficacy of such molecules and also improve the physical and chemical properties of ceramide compounds, for example, enhance the antioxidant activity of ceramide compounds. BRIEF DESCRIPTION OF THE DRAWINGS

[0051] Figure 1 The test results of the antioxidant activity of the ceramide compound in Example 10 are shown in FIG. DETAILED DESCRIPTION

[0052] The present invention will be further described below in conjunction with specific embodiments.

[0053] All reactions were carried out under a nitrogen atmosphere. Unless otherwise stated, chemicals were purchased from commercial products and were not further purified. Dichloromethane and tetrahydrofuran used in the experiments were anhydrous solvents. Thin layer chromatography (TLC) used 60F254 silica gel plates. Silica gel column chromatography used Qingdao Ocean Silica Gel (particle size 0.040-0.063 mm). TLC color development used UV light (254 nm) or iodine. NMR spectra were characterized using a Bruker DPX 400 nuclear magnetic resonance instrument, 1 H NMR is at 400 MHz, the solvent is deuterated methanol, deuterated DMSO or deuterated tetrahydrofuran, with tetramethylsilane (TMS) as the internal standard. The unit of chemical shift is ppm, and the unit of coupling constant is Hz. 1 In H NMR, δ represents chemical shift, s represents singlet, d represents doublet, t represents triplet, q represents quartet, and m represents multiplet.

[0054] DCC refers to N,N'-dicyclohexylcarbodiimide, THF refers to tetrahydrofuran, DCM refers to dichloromethane, EDCI refers to 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, and HOBT refers to 1-hydroxybenzotriazole.

[0055] General Synthesis Method of Ceramide Compounds

[0056] Method A

[0057]

[0058] Step 1: Compound S1 (50 mmol), N-hydroxysuccinimide (60-75 mmol, preferably 60 mmol), and DCC (60-75 mmol, preferably 60 mmol) were placed in a 250 mL round-bottom flask, 100 mL of tetrahydrofuran was added, and then stirred at room temperature for 4-24 hours, and detected by TLC until compound S1 completely disappeared. Post-treatment: Filter the reaction system, collect the filtrate and use it directly in the next step reaction.

[0059] Step 2: Add sphingoid base (40-50 mmol, preferably 45 mmol) to the filtrate of the previous step, then stir at room temperature for 24-72 hours, and detect by TLC until the sphingoid base disappears completely. Post-treatment: add saturated sodium bicarbonate aqueous solution to quench. Separate the organic layer, dry, filter and concentrate in vacuo. The residue thus obtained is purified by silica gel column to obtain the product (50-70% yield).

[0060] Method B

[0061]

[0062] Compound S1 (50 mmol), EDCI (60-75 mmol, preferably 60 mmol), HOBT (60-75 mmol, preferably 60 mmol) are placed in a 250 mL round-bottom flask, 100 mL of dichloromethane is added, and then stirred at room temperature for 1 hour, then sphingoid base (40-50 mmol, preferably 45 mmol) is added to the reaction system, and stirred at room temperature for 24-72 hours until the sphingoid base disappears completely. Post-treatment: quench with water. Separate the organic layer, dry, filter and concentrate in vacuo. The residue thus obtained is purified by silica gel column to obtain the product (50-65% yield).

[0063] Example 1

[0064] The condensation product of ferulic acid and phytosphingosine by method A.

[0065]

[0066] 1 H NMR (400 MHz, Methanol-d 4 )δ7.45(d,J=15.6Hz,1H),7.12(d,J=2.0Hz,1H),7.03(dd,J=8.3,1.9Hz,1H),6.79(d,J=8.1Hz,1 H),6.53(d,J=15.7Hz,1H),4.23(q,J=5.3Hz,1H),3.88(s,3H),3.80(qd,J=11.2,5.0Hz,2H),3.64 (t,J=5.8Hz,1H),3.56(ddd,J=8.9,5.9,2.5Hz,1H),1.68(dtd,J=12.9,7.2,6.2,3.5Hz,1H),1.5 3(d,J=8.4Hz,1H),1.43(ddd,J=12.4,8.1,3.7Hz,1H),1.36–1.16(m,23H),0.89(t,J=6.7Hz,3H).

[0067] Example 2

[0068] The product of the condensation of ferulic acid and sphingosine by method A.

[0069]

[0070] 1 H NMR (400 MHz, Methanol-d 4)δ7.43(d,J=15.7Hz,1H),7.12(d,J=2.0Hz,1H),7.03(dd,J=8.2,2.0Hz,1H),6.79(d,J=8. 1Hz,1H),6.50(d,J=15.7Hz,1H),5.75–5.62(m,1H),5.55–5.44(m,1H),4.35(t,J=7.0Hz,1 H),4.12(t,J=7.4Hz,1H),4.01–3.94(m,1H),3.88(s,3H),3.78–3.68(m,1H),2.54–2.45(m ,1H),2.30–2.16(m,1H),2.02(p,J=7.2Hz,2H),1.37–1.10(m,20H),0.89(t,J=6.9Hz,3H).

[0071] Example 3

[0072] The product of the condensation of ferulic acid and dihydrosphingosine by method A.

[0073]

[0074] 1 H NMR (400 MHz, Methanol-d 4 )δ7.48(d,J=15.7Hz,1H),7.16(d,J=2.0Hz,1H),7.06(dd,J=8.2,1.9Hz,1H),6.82(d,J=8.2Hz,1H),6.55(d,J=15.7Hz,1H),4.0 1–3.93(m,1H),3.91(s,3H),3.79(d,J=5.0Hz,2H),3.77–3.71(m,1H),1.92–1.85(m,2H),1.38–1.23(m,26H),0.96–0.87(m,3H).

[0075] Example 4

[0076] The condensation product of trans-cinnamic acid and phytosphingosine by method A.

[0077]

[0078] 1 H NMR (400 MHz, Methanol-d 4)δ7.60–7.50(m,3H),7.42–7.31(m,3H),6.71(d,J=15.8Hz,1H),4.25(td ,J=5.7,4.2Hz,1H),3.83(dd,J=11.3,4.3Hz,1H),3.77(dd,J=11.3,5.8Hz ,1H),3.64(t,J=5.8Hz,1H),3.56(ddd,J=9.1,5.9,2.5Hz,1H),1.75–1.61 (m,1H),1.53(d,J=9.1Hz,1H),1.48–1.16(m,24H),0.90(t,J=6.8Hz,3H).

[0079] Example 5

[0080] The product of the condensation of royal jelly acid and phytosphingosine by method A.

[0081]

[0082] 1 H NMR (600MHz, Methanol-d 4 )δ6.81(dt,J=15.3,7.0Hz,1H),6.02(dt,J=15.3,1.5Hz,1H),4.19(td,J=5.8,4.3Hz,1H),3.81(dd,J=11.3,4.4Hz,1H),3.76(dd,J=11.2,5. 8Hz,1H),3.62(t,J=5.8Hz,1H),3.58–3.53(m,3H),2.23(qd,J=7.2,1.6Hz,2H),1.57–1.49(m,5H),1.39–1.29(m,32H),0.92(t,J=7.0Hz,3H).

[0083] Example 6

[0084] Condensation product of retinoic acid and phytosphingosine by method B

[0085]

[0086] 1 H NMR (400 MHz, Methanol-d 4)δ7.01(dd,J=15.0,11.4Hz,1H),6.32(dd,J=23.7,15.6Hz,2H),6.21–6.09(m,2H),5.93(s,1H), 4.17(td,J=5.7,4.4Hz,1H),3.86–3.72(m,2H),3.64(t,J=5.8Hz,1H),3.61–3.53(m,1H),2.33(d, J=1.1Hz,3H),2.06(t,J=6.5Hz,2H),2.01(d,J=1.1Hz,3H),1.73(d,J=1.0Hz,3H),1.66(ddp,J=8. 8,5.6,2.9Hz,3H),1.54–1.47(m,3H),1.30(d,J=4.9Hz,25H),1.05(s,6H),0.92(t,J=6.8Hz,3H).

[0087] Example 7

[0088] Condensation product of sinapinic acid and phytosphingosine by method B

[0089]

[0090] 1 H NMR (400 MHz, Methanol-d 4 )δ7.47(d,J=15.6Hz,1H),6.88(s,2H),6.58(d,J=15.6Hz,1H),4.26(td,J=5.6,4.3Hz,1H),3.89(s,6H),3.85–3.78(m,2H),3.67(t,J=5 .8Hz,1H),3.59(ddd,J=9.0,5.9,2.6Hz,1H),1.77–1.61(m,2H),1.61–1.43(m,2H),1.29(dd,J=11.8,5.4Hz,22H),0.92(t,J=6.8Hz,3H).

[0091] Example 8

[0092] The condensation product of caffeic acid and phytosphingosine by method B.

[0093]

[0094] 1 H NMR (400 MHz, Methanol-d 4)δ7.53(d,J=15.9Hz,1H),7.02(d,J=2.1Hz,1H),6.92(dd,J=8.2,2.1Hz,1H),6.76(d,J=8.2H z,1H),6.24(d,J=15.9Hz,1H),4.20(q,J=5.3Hz,1H),3.78(qd,J=11.2,5.0Hz,2H),3.62(t,J= 6.0Hz,1H),3.50(ddd,J=8.9,6.0,2.4Hz,1H),1.64(dtd,J=12.9,7.2,6.2,3.5Hz,1H),1.51(d ,J=8.4Hz,1H),1.43(ddd,J=12.4,8.1,3.7Hz,1H),1.36–1.16(m,23H),0.89(t,J=6.7Hz,3H).

[0095] Example 9

[0096] The condensation product of sorbic acid and phytosphingosine by method B.

[0097]

[0098] 1 H NMR (400 MHz, Methanol-d 4 )δ7.70(td,J=6.8,3.8Hz,1H),7.32–7.23(m,1H),7.14(dd,J=15.1,10.6Hz,1H),6.25(ddd,J=15.1 ,10.6,1.7Hz,1H),6.13(dq,J=15.2,6.5Hz,1H),6.00(d,J=15.1Hz,1H),4.20(td,J=5.7,4.3Hz,1H ),3.79(qd,J=11.2,5.1Hz,2H),3.62(t,J=5.8Hz,1H),3.55(ddd,J=8.9,5.9,2.5Hz,1H),1.86(dd, J=6.6,1.4Hz,3H),1.74–1.61(m,1H),1.54(d,J=9.0Hz,1H),1.39–1.30(m,24H),0.95–0.86(m,3H).

[0099] Example 10

[0100] The antioxidant activity of the ceramide compounds prepared in Examples 1, 2, 3 and 5 was tested by the antioxidant-ABTS method.

[0101] Experimental principle: The method of evaluating the antioxidant capacity of samples using ABTS (2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonicacid)diammonium salt) was first proposed by Miller et al. (1993), and the method currently used is generally the improved method by Re et al. (1999). This method uses the fact that ABTS can be oxidized by a series of compounds such as potassium persulfate, hydrogen peroxide, and manganese dioxide to generate blue-green ABTS+ cationic free radicals with a maximum absorption peak at 734nm. Under the action of antioxidants, ABTS+ is reduced to colorless ABTS. By measuring the absorbance value at 734nm, the antioxidant capacity of the reactant can be determined.

[0102] ABTS free radical scavenging test for antioxidant performance: 3 mL of ABTS aqueous solution (12 mmol / L) and 3 mL of potassium persulfate solution (2.45 mmol / L) were mixed evenly and kept stable at room temperature in the dark for 12 to 16 hours. DMSO was used as the dilution medium, and the appropriate dilution ratio was selected to adjust the absorbance of the potassium persulfate solution at 734 nm to 0.700 ± 0.025. Samples of different concentrations were added and shaken, kept in the dark for 10 minutes, and the absorbance was measured at 734 nm.

[0103] The results are as follows Figure 1 As shown, control 1 is ceramide 3B and control 2 is ceramide 3.

[0104] It can be seen that the compounds of the present invention have better antioxidant effects than existing ceramide compounds, and the scavenging rate of free radicals is generally 15-75% higher.

[0105] The above is only a specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any changes or substitutions that can be easily thought of by a person skilled in the art within the technical scope disclosed by the present invention should be included in the protection scope of the present invention. Therefore, the protection scope of the present invention should be based on the protection scope of the claims.

Claims

1. A ceramide compound having a structure of general formula I or an enantiomer or diastereomer of general formula I: in, R 1 Selected from the residues after condensation of royal jelly acid or sorbic acid, The R 2 Select one of the following structures:

2. The ceramide compound according to claim 1, characterized in that Select one of the following compounds:

3. A method for preparing the ceramide compound according to claim 1 or 2, characterized in that: The following steps are involved: Compound S1 reacts with N-hydroxysuccinimide and a condensation agent to obtain compound S2; Compound S2 reacts with sphingoid base to obtain compound I; R 1 is the residue after condensation of royal jelly acid, R 2 As defined in claim 1 or 2.

4. The method according to claim 3, characterized in that The molar ratio of the compound S1, N-hydroxysuccinimide, condensing agent and sphingoid base is 1:(1-2):(1-2):(0.8-1); the condensing agent is DCC; and the solvent of the reaction is THF.

5. A method for preparing the ceramide compound according to claim 1 or 2, characterized in that: The following steps are involved: Compound S1 reacts with a sphingoid base and a condensing agent to obtain compound I; R 1 is the residue after condensation of sorbic acid, R 2 As defined in claim 1 or 2.

6. The method according to claim 5, characterized in that The condensing agent includes EDCI and HOBT, and the molar ratio of the compound S1, EDCI, HOBT and sphingoid base is 1:(1-2):(1-2):(0.8-1); the solvent of the reaction is DCM.

7. Use of the ceramide compound according to claim 1 or 2 in the preparation of an antioxidant.

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