Amino acid ceramide derivative and method for synthesizing the same

Abstract

The application provides a novel ceramide derivative based on amino acid synthesis and a preparation method thereof, wherein the ceramide derivative is synthesized by introducing a saturated or unsaturated fatty acid into an amino acid. The ceramide derivative has the ability to repair and maintain a skin barrier, wherein the raw material amino acid is low in price and can be used as a raw material for industrial production, and the amino acid has a large number of applications in food, daily chemicals and the like, is high in activity, is good in skin affinity, and can be added to cosmetics to play a high-efficiency role after testing.

Description

Technical Field

[0001] This invention belongs to the field of compound synthesis technology, and more specifically, relates to an amino acid-based ceramide derivative and its synthesis method. Background Technology

[0002] Ceramides, unsaturated fatty acids added to daily cosmetics, are naturally abundant in the cytoplasm of skin cells and play an important role in maintaining the skin barrier. Ceramides and their analogues are used to rapidly restore skin hydration and repair the barrier, making them an important ingredient in high-end cosmetics.

[0003] The chemical route mainly synthesizes ceramide-like substances, which have similar structures and functions to ceramides and can be used in cosmetics. Currently, several ceramide-like substances have been successfully synthesized. Amorepacific has produced a new type of ceramide-like substance (CN104854081A), which has improved its solubility and stability. The technical route is to dissolve tris(hydroxymethyl)aminomethane in dimethylformaldehyde (DMF), then dissolve it in triethylamine (Et3N), and after 30 minutes, add palmitoyl chloride and introduce fatty acids through a series of reactions. The reaction formula is shown in (1). This method has a relatively stable amidation process, but the two-step addition, especially the second step of amide and alcohol condensation, easily generates multi-substituted ceramide-like derivatives, resulting in reduced yield and product efficacy, and higher cost.

[0004]

[0005] In 2006, Sun Yat-sen Memorial Hospital of Sun Yat-sen University published a patent (CN110041221A) for a ceramide analogue, its preparation method and application. This method synthesized a new class of fatty acid hydroxy fatty acid esters (FAFAs) by reacting FA15 with 9-hydroxyoctadecanoic acid, and synthesized five novel ceramide analogues (FAFACerAE) by reacting sphingosine with the corresponding five FAFAs. This method is lengthy, has a low yield, and is costly, making it unsuitable for industrial production. Its reaction formula is shown in (2):

[0006]

[0007] In 2022, Suzhou Element Collection Chemical Industry Co., Ltd. published a patent (CN115850110A): a method for synthesizing a ceramide-like compound containing a glycerol ester structure. The reaction uses a long-chain hydrocarbon acyl dicarboxylic acid as the starting material. Although the reaction yield is high and the product has good activity, the cost is inevitably high and it is difficult to industrialize. The reaction formula is shown in (3): Summary of the Invention

[0008] The purpose of this invention is to overcome the problems existing in the prior art and provide an amino acid-based ceramide derivative and its synthesis method. The amino acid-based ceramide analog provided by this invention has excellent ability to maintain skin moisture and repair the skin barrier.

[0009] The objective of this invention and the technical problem it solves are achieved by the following technical solutions.

[0010] One aspect of the present invention provides an amino acid-based ceramide derivative of formula (I),

[0011]

[0012] Or its tautomers, or pharmaceutically acceptable salts, prodrugs, hydrates or solvates thereof of said derivatives or tautomers, wherein:

[0013] R1 and R2 are independently C0-C 99 Saturated or unsaturated fatty acid chains;

[0014] R3 is a structure containing hydroxyl, carboxyl, amino, sulfonyl, aromatic, saturated or unsaturated hydrocarbon groups.

[0015] Preferably, R1 and R2 are C 15 H 31 R3 is CH3.

[0016] Another aspect of the present invention provides a method for preparing the aforementioned amino acid-based ceramide derivatives, the method comprising the following steps:

[0017] S1: Dissolve amino acids in DMF, add alkali, add acyl chloride dropwise under nitrogen protection, stir at room temperature for 1-48 h, then add water and stir for 0.5-48 h, finally filter and dry to obtain the amidated product;

[0018] S2: Dissolve the above-obtained amidation product in DMF, add alkali and halogenated hydrocarbon, stir at 50-80℃ for 1-48h, then add water and stir for 0.5-48h, finally filter and dry to obtain the final product.

[0019] Preferably, the chemical formula of the reaction in the above method is:

[0020]

[0021] Preferably, the amino acids in step S1 include serine, valine, threonine and their derivatives.

[0022] Preferably, the alkali mentioned in step S1 is a commonly used organic or inorganic alkali in the art, including but not limited to triethylamine, sodium carbonate, potassium carbonate, potassium bicarbonate, sodium bicarbonate, sodium hydroxide, and potassium hydroxide.

[0023] Preferably, the acyl chloride in step S1 is a long-chain acyl chloride, including but not limited to decaacyl chloride, dodecyl chloride, hexadecyl chloride, tetradecyl chloride, octadecyl chloride, icosyl chloride, and docosyl chloride.

[0024] Preferably, the alkali mentioned in step S2 is a commonly used organic or inorganic alkali in the art, including but not limited to triethylamine, sodium carbonate, potassium carbonate, potassium bicarbonate, sodium bicarbonate, sodium hydroxide, and potassium hydroxide.

[0025] Preferably, the haloalkane in step S2 is a brominated long-chain alkane, including but not limited to bromodecane, bromododecane, bromohexadecane, bromooctadecane, bromoeicosane, bromodocosahexadecane, and bromotetradecane.

[0026] Another aspect of the present invention provides the use of the aforementioned amino acid ceramide derivative, or its tautomer, or a pharmaceutically acceptable salt, prodrug, hydrate, or solvate thereof of the derivative or tautomer in the preparation of daily chemical products or medicaments for treating skin diseases.

[0027] Another aspect of the present invention provides the use of an amino acid ceramide derivative, or a tautomer thereof, prepared by the aforementioned method, or a pharmaceutically acceptable salt, prodrug, hydrate thereof, or solvate thereof, of the derivative or tautomer in the preparation of a daily chemical product or a dermatological treatment drug.

[0028] By employing the above technical solution, the present invention has at least the following advantages:

[0029] (1) This invention synthesizes a novel amino acid-type ceramide analog by introducing saturated or unsaturated alkane esters onto amino acids. The synthetic method of the amino acid-type ceramide analog disclosed in this invention is simple and has a high overall yield (over 80%).

[0030] (2) The amino acids used in this invention have the advantages of low price and wide availability. They can be used as raw materials for industrial production. Amino acids have a wide range of applications in food, daily chemicals and other fields. By introducing saturated and unsaturated fatty acids into amino acids, ceramide derivatives are synthesized. They have high activity and good skin affinity. After testing, they can be added to cosmetics to play a high-efficiency role.

[0031] (3) The amino acid-type ceramide analog obtained by the method of the present invention has a good ability to maintain skin moisture and repair the skin barrier, suggesting that it may have a wide range of applications in daily chemical products or drugs for treating skin diseases.

[0032] The above description is merely an overview of the technical solution of the present invention. In order to better understand the technical means of the present invention and to implement it in accordance with the contents of the specification, the preferred embodiments of the present invention are described in detail below. Detailed Implementation

[0033] To make the technical means, creative features, achieved objectives, and effects of this invention readily understandable, the technical solutions in the embodiments of this invention will be clearly and completely described below in conjunction with the embodiments of this invention. Obviously, the described embodiments are merely some embodiments of this invention, and not all embodiments. Based on the embodiments of this invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this invention.

[0034] The ceramide derivative, or its tautomer, or a pharmaceutically acceptable salt, prodrug, hydrate, or solvate thereof according to the present invention, has the structural formula shown in (I):

[0035]

[0036] R1 and R2 are independently C0-C99 saturated or unsaturated aliphatic chains;

[0037] R3 is a structure containing hydroxyl, carboxyl, amino, sulfonyl, aromatic, saturated or unsaturated hydrocarbon groups.

[0038] Preferably, R1 and R2 are C 15 H 31 R3 is CH3.

[0039] The method for preparing the above compounds is as follows:

[0040] S1: Dissolve amino acids in DMF, add alkali, add acyl chloride dropwise under nitrogen protection, stir at room temperature for 1-48 h, then add water and stir for 0.5-48 h, finally filter and dry to obtain the amidated product;

[0041] S2: Dissolve the above-obtained amidation product in DMF, add alkali and halogenated hydrocarbon, stir at 50-80℃ for 1-48h, then add water and stir for 0.5-48h, finally filter and dry to obtain the final product.

[0042] The chemical formula for the reaction described above is:

[0043]

[0044] Example 1:

[0045] 1 mol of threonine was dissolved in 200 mL of DMF at room temperature. 1.5 mol of triethylamine was added, followed by 1 mol of hexadecyl chloride, which was added dropwise over 1 hour. The mixture was stirred at room temperature for 3 hours. Then, 1 L of water was added and stirred for 0.5 hours. The mixture was filtered, and the filter cake was dried to obtain the amidated product with a yield of 95%.

[0046] 0.95 mol of the amidated product was dissolved in 190 mL of DMF at room temperature. 1.43 mol of triethylamine and 0.95 mol of hexadecane bromide were added, and the mixture was reacted at 50 °C for 3 h. Then, 1 L of water was added, and the mixture was stirred for 0.5 h. The mixture was filtered, and the filter cake was dried to obtain a threonine-type ceramide analog. Yield: 98%. Overall yield: 93.1%.

[0047] Example 2:

[0048] 1 mol of threonine was dissolved in 200 mL of DMF at room temperature. 1.5 mol of triethylamine was added, followed by 1 mol of tetradecyl chloride, which was added dropwise over 1 hour. The mixture was stirred at room temperature for 3 hours. 1 L of water was added, and the mixture was stirred at room temperature for 0.5 hours. The mixture was then filtered, and the filter cake was dried to obtain the amidated product with a yield of 90%.

[0049] 0.90 mol of the amidated product was dissolved in 180 mL of DMF at room temperature. 1.35 mol of potassium carbonate and 0.90 mol of bromooctadecane were added, and the mixture was reacted at 50 °C for 3 h. Then, 1 L of water was added, and the mixture was stirred for 0.5 h. The mixture was filtered, and the filter cake was dried to obtain a threonine-type ceramide analog. Yield: 95%. Overall yield: 85.5%.

[0050] Example 3:

[0051] 1 mol of serine was dissolved in 200 mL of DMF at room temperature. 1.5 mol of sodium carbonate was added, and 1 mol of hexadecyl chloride was added dropwise over 1 hour. The mixture was stirred at room temperature for 3 hours. Then, 1 L of water was added and stirred for 0.5 hours. The mixture was filtered, and the filter cake was dried to obtain the amidated product with a yield of 90%.

[0052] 0.90 mol of the amidated product was dissolved in 180 mL of DMF at room temperature. 1.35 mol of triethylamine and 0.90 mol of hexadecane bromide were added, and the mixture was reacted at 60 °C for 3 h. Then, 1 L of water was added, and the mixture was stirred for 0.5 h. The mixture was filtered, and the filter cake was dried to obtain a serine-type ceramide analog. Yield: 95%. Overall yield: 85.5%.

[0053] The NMR data of the obtained product are as follows:

[0054] H NMR(500MHz; CDCl3),d 0.9(t,6H,CH3); 1.3(s,50H,CH2); 1.6(m,4H,b-CH2); 2.2(t,2H,CH2CONH); 3.9(d,2H ,HOCH2CHNH);4.15(t,2H,CH2OOC);4.65(m,1H,CH6(NH));6.4(d,1H,NH6).Elemental analysis,calculated for C35H69NO4,C,74.07;H,12.17;N,2.47;found,C,74.03;H,11.94;N,2.43.

[0055] Example 4:

[0056] 1 mol of serine was dissolved in 200 mL of DMF at room temperature. 1.5 mol of potassium bicarbonate was added, and 1 mol of hexadecyl chloride was added dropwise over 1 hour. The mixture was stirred at room temperature for 3 hours. Then, 1 L of water was added and stirred for 0.5 hours. The mixture was filtered, and the filter cake was dried to obtain the amidated product with a yield of 98%.

[0057] 0.98 mol of the amidated product was dissolved in 198 mL of DMF at room temperature. 0.98 mol of triethylamine and 0.98 mol of bromodocoane were added, and the mixture was reacted at 80 °C for 3 h. Then, 1 L of water was added, and the mixture was stirred for 0.5 h. The mixture was filtered, and the filter cake was dried to obtain a serine-type ceramide analog. Yield: 90%. Overall yield: 88.2%.

[0058] Example 5:

[0059] 1 mol of tyrosine was dissolved in 200 mL of DMF at room temperature. 1.5 mol of sodium bicarbonate was added, followed by the dropwise addition of 1 mol of hexadecyl chloride over 1 hour. The mixture was stirred at room temperature for 3 hours. Then, 1 L of water was added, and the mixture was stirred for 0.5 hours. The mixture was then filtered, and the filter cake was dried to obtain the amidated product with a yield of 91%.

[0060] 0.91 mol of the amidated product was dissolved in 182 mL of DMF and 1.37 mol of triethylamine at room temperature. 0.91 mol of tetradecane was added, and the mixture was reacted at 60 °C for 3 h. Then, 1 L of water was added, and the mixture was stirred for 0.5 h. The mixture was filtered, and the filter cake was dried to obtain a tyrosine-type ceramide analog. Yield: 95%. Overall yield: 86.45%.

[0061] Example 6:

[0062] Take 1 mol of tyrosine, dissolve it in 200 mL of DMF at room temperature, add 1.5 mol of sodium hydroxide, and add 1 mol of octadecyl chloride dropwise over 1 hour. Stir the reaction at room temperature for 3 hours, add 1 L of water, stir for 0.5 hours, filter, dry the filter cake to obtain the amidated product, yield: 95%.

[0063] 0.95 mol of the amidated product was dissolved in 190 mL of DMF and 1.425 mol of triethylamine at room temperature. 0.95 mol of bromooctadecane was added, and the mixture was reacted at 70 °C for 3 h. Then, 1 L of water was added, and the mixture was stirred for 0.5 h. The mixture was filtered, and the filter cake was dried to obtain a tyrosine-type ceramide analog. Yield: 95%. Overall yield: 90.25%.

[0064] Experimental Example: Performance Testing of Ceramide-like Compounds

[0065] Experimental methods: The ceramide-like compounds synthesized in Examples 1-6 were subjected to performance testing experiments. Specific testing methods included:

[0066] 1) Determine the safe concentration of keratinocytes based on the MTT assay principle;

[0067] 2) The inhibition rate of collagenase and hyaluronidase was determined by enzyme activity assay;

[0068] 3) Using qPCR technology, fluorescent dyes or fluorescently labeled specific probes are used to label and track the fluorescent quantitative products, monitor the reaction process in real time, and detect their expression of skin barrier-related proteins (K10, IVL, LOR, FASN) and inflammatory factors (IL-6).

[0069] Experimental results are shown in Table 1 below:

[0070] Table 1 Summary of performance tests of the ceramide-like compounds synthesized in Examples 1-6

[0071]

[0072] As shown in Table 1, the safe concentration of ceramide-like keratinocytes obtained by the method according to the present invention meets the requirements. It has good effects on collagenase inhibition rate, hyaluronidase inhibition rate, expression of skin barrier-related proteins and inflammatory factor proteins, indicating that it has good anti-aging, soothing and skin barrier repair capabilities, suggesting that it may have a wide range of application prospects in daily chemical products or drugs for treating skin diseases.

[0073] In summary, the novel ceramide derivatives based on amino acid synthesis provided by this invention have the ability to repair and maintain the skin barrier. Amino acids are inexpensive and can be used as raw materials for industrial production. Furthermore, amino acids have numerous applications in food and daily chemical products. Introducing saturated and unsaturated fatty acids into amino acids to synthesize ceramide derivatives results in high activity and good skin affinity. Testing has shown that these derivatives can be added to cosmetics to exert a highly effective effect.

[0074] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art can make some modifications or alterations to the methods and techniques disclosed above without departing from the scope of the present invention to create equivalent embodiments. Any simple modifications, equivalent changes and alterations made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the scope of the present invention.

Claims

1. A method for preparing a threonine-type ceramide analog, characterized in that, Includes the following steps: 1 mol of threonine was dissolved in 200 mL of DMF at room temperature. 1.5 mol of triethylamine was added, followed by the dropwise addition of 1 mol of hexadecyl chloride over 1 hour. The mixture was stirred at room temperature for 3 hours. Then, 1 L of water was added, and the mixture was stirred for 0.5 hours. The mixture was filtered, and the filter cake was dried to obtain the amidated product with a yield of 95%. 0.95 mol of the amidation product was dissolved in 190 mL of DMF at room temperature, 1.43 mol of triethylamine was added, and 0.95 mol of hexadecane bromide was added. The mixture was reacted at 50 °C for 3 h, then 1 L of water was added, and the mixture was stirred for 0.5 h. The mixture was filtered, and the filter cake was dried to obtain a threonine-type ceramide analog. The yield was 98%, and the overall yield was 93.1%.

2. The use of the amino acid ceramide compound or its tautomer or a pharmaceutically acceptable salt thereof obtained by the preparation method of claim 1 in the preparation of daily chemical products or medicaments for treating skin diseases.

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