A highly efficient transdermal absorption arbutin analogue and a preparation method thereof

Abstract

The application discloses a preparation method of arbutin analogues with high transdermal absorption efficiency, and is characterized in that the method comprises the following steps: under the action of a solvent and a catalyst, compound I and compound II are subjected to esterification to obtain compound III, wherein the compound III is the arbutin analogue. The arbutin (compound I) is used as a raw material, the structure of the arbutin is modified through esterification, the transdermal performance of the arbutin is improved, the bioavailability is increased, and finally the whitening effect is improved.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of organic matter and intermediate synthesis, and particularly relates to a high-efficiency transdermal absorption arbutin analogue and a preparation method thereof. BACKGROUND

[0002] Arbutin is a natural active substance isolated from various plants. In the 1990s, it was first introduced by Shiseido Cosmetics Corporation of Japan as a cosmetic whitening agent. It is another whitening agent after gamma-cysteine and kojic acid. It not only has fading effect on skin freckles, age spots and chloasma, but also has good curative effect on skin moisturizing, healing after skin burns and acne. At present, the developed countries' whitening skin care market has almost been monopolized by arbutin.

[0003] Yang et al. (Bioresour Technol. 2010, 101(1): 1-5.) found that the cell membrane permeability greatly limits the absorption of arbutin when studying the bioavailability of arbutin in the human body. SUMMARY

[0004] In view of the above problems, the present application provides a high-efficiency transdermal absorption arbutin analogue and a preparation method thereof. The present application uses arbutin as raw material, modifies the structure of arbutin through esterification reaction, improves the transdermal performance, increases the bioavailability, and finally achieves the effect of improving whitening.

[0005] The purpose of the present application and the technical problems thereof are realized by adopting the following technical solutions.

[0006] One aspect of the present application provides a preparation method of a high-efficiency transdermal absorption arbutin analogue, which comprises the following steps: compounds I and II are subjected to esterification reaction under the action of a solvent and a catalyst to obtain compound III, and the synthesis route is as follows:

[0007]

[0008] In the formula, X represents halogen; and R represents alkyl.

[0009] Preferably, in the compound II, X represents one of F, Cl, Br and I; and R represents C1-C30 alkyl.

[0010] More preferably, in the compound II, X represents Cl.

[0011] Preferably, the solvent is selected from one or a mixture of several of dimethyl sulfoxide, N,N-dimethylformamide, acetonitrile, dichloromethane, chloroform and carbon tetrachloride.

[0012] Preferably, the solvent is N,N-dimethylformamide.

[0013] Preferably, the catalyst is selected from one or a mixture of several of pyridine, triethylamine, diethylamine, isopropylamine, diisopropylethylamine, N-methylmorpholine, tetramethylethylenediamine.

[0014] Preferably, the catalyst is pyridine.

[0015] Preferably, the esterification reaction temperature is 20-100℃.

[0016] Preferably, the esterification reaction temperature is 60-80℃.

[0017] Preferably, the esterification reaction time is 1-24h.

[0018] Preferably, the esterification reaction time is 3-8h.

[0019] By the above technical solution, the present application has at least the following advantages: the present application uses arbutin (compound I) as raw material, and the product is obtained through esterification, separation and purification. After specific esterification, the liposolubility of arbutin is increased, and the transdermal absorption performance is greatly enhanced. The arbutin (compound I), compound II, catalyst and solvent used in the present application are easy to obtain and low in price, so that the production cost is low. The esterification reaction condition of the present application is mild, the reaction time is short, the production cycle can be effectively reduced, the production efficiency is improved, and the production energy consumption is reduced. The post-treatment process of the present application is simple and easy to operate. The obtained arbutin analogue has obviously enhanced liposolubility, improved transdermal absorption performance, and significantly improved whitening effect.

[0020] The above description is only a summary of the technical solution of the present application. In order to more clearly understand the technical means of the present application, and to implement the content of the description, the following preferred embodiments of the present application and the drawings are described in detail. BRIEF DESCRIPTION OF DRAWINGS

[0021] Figure 1 The liquid chromatogram of the high-efficiency transdermal absorption arbutin analogue obtained according to the preparation method of embodiment 1 of the present application is shown;

[0022] Figure 2 The schematic diagram of Franz diffusion cell is shown;

[0023] Figure 3 The transdermal diffusion test result comparison diagram of the high-efficiency transdermal absorption arbutin analogue obtained according to the preparation method of embodiment 1 of the present application and the arbutin of comparative example 1 on artificial skin membrane is shown;

[0024] Figure 4A comparison chart of the results of the cell melanin inhibition test of the high-efficiency transdermally absorbed arbutin analog obtained according to the preparation method of Embodiment 1 of the present application and arbutin of Comparative Embodiment 1 is shown. DETAILED DESCRIPTION

[0025] In order to make the technical means, creative features, purposes and effects realized by the present application easy to understand, the technical solutions in the embodiments of the present application will be described clearly and completely below in conjunction with the embodiments of the present application and the accompanying drawings. Obviously, the described embodiments are only a part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without any creative effort fall within the scope of protection of the present application.

[0026] Example 1

[0027] The arbutin analog in the present embodiment is prepared according to the following method:

[0028] A 1000 mL reaction bottle is sequentially added with 50 g of compound I, 400 mL of N,N-dimethylformamide, 20 mL of pyridine and 50 g of compound II, heated to 60°C for 4 h of reaction, after the reaction is completed, cooled to room temperature, drop 400 mL of purified water, filter, filter cake is blown at 50°C for 8 h of drying to obtain 68 g of arbutin analog (compound III) (calculated yield is 69.5%).

[0029] The above reaction chemical equation is:

[0030]

[0031] In the embodiments of the present application, high performance liquid chromatography is used to determine the purity of the product, and the method and conditions are as follows:

[0032] Chrompock C18 (250 mm x 4.6 mm, 5 um) is used as the chromatographic column;

[0033] The mobile phase is methanol: 0.1 trifluoroacetic acid (TFA) aqueous solution = 10:90;

[0034] The flow rate is 0.8 ml / min;

[0035] The injection volume is 20 uL;

[0036] The detection wavelength is UV 280 nm;

[0037] The column temperature is 40°C.

[0038] Figure 1 The liquid chromatogram of the obtained arbutin analog (compound III) is shown in Figure 1It can be seen that the product obtained in this embodiment has high purity.

[0039] Example 2

[0040] The arbutin analogue in this embodiment was prepared according to the following method:

[0041] Compound I 50g, dimethyl sulfoxide 400mL, pyridine 20mL and compound II 50g were added sequentially to a 1000mL reaction flask. The mixture was heated to 70℃ and reacted for 6h. After the reaction was completed, the mixture was cooled to room temperature, 400mL of purified water was added dropwise, and the mixture was filtered. The filter cake was dried at 50℃ for 8h under forced air to obtain 59g of arbutin analog (compound III) (the yield was calculated to be 60.2%).

[0042] The chemical equation for the above reaction is:

[0043]

[0044] Example 3

[0045] The arbutin analogue in this embodiment was prepared according to the following method:

[0046] Compound I 50g, dichloromethane 400mL, triethylamine 20mL and compound II 50g were added sequentially to a 1000mL reaction flask. The mixture was heated to 40℃ and reacted for 8h. After the reaction was completed, the mixture was concentrated to dryness, 400mL of purified water was added and stirred for 0.5h, filtered, and the filter cake was dried at 50℃ for 8h to obtain 39g of arbutin analog (compound III) (the yield was calculated to be 39.7%).

[0047] The chemical equation for the above reaction is:

[0048]

[0049] Comparative Example 1

[0050] Commercially available arbutin (West Asia Reagent, catalog number: A12023)

[0051] Test Example 1 Cell Transdermal Absorption Test

[0052] Experimental subjects: Arbutin analogue (compound III) of Example 1 and arbutin of Comparative Example 1.

[0053] Experimental Methods: Transdermal diffusion experiments were conducted using Strat-M artificial skin membranes. Specifically, the sample was dissolved in ethanol, and ethanol was used as the solvent in the receiving cell. The principle is explained below. Figure 2 As shown. The results are attached. Figure 3 .

[0054] Figure 3 A comparison chart of the results of the transdermal diffusion test of the high-efficiency transdermally absorbed arbutin analog according to the preparation method of Example 1 of the present application and the artificial skin membrane of arbutin of Comparative Example 1 is shown. From the chart, it can be seen that the sample concentration of the arbutin analog of Example 1 (Compound III) and arbutin of Comparative Example 1 through the artificial membrane gradually increased with the extension of time (1 h-5 h), and the transdermal performance of the arbutin analog of Example 1 (Compound III) was obviously stronger than that of arbutin of Comparative Example 1. Figure 3

[0055] Test Example 2 Cell Melanin Inhibition Test

[0056] Experimental subjects: arbutin analog of Example 1 (Compound III) and arbutin of Comparative Example 1.

[0057] Experimental method: The principle is that the mouse B16 melanocyte can release melanin after alkaline lysis, and the melanin of the cells after 72 h of action of different experimental samples (same concentration 1%, 2%, 3%) is extracted, and the high and low of the inhibition effect is reacted by OD.

[0058] Specific method is as follows:

[0059] I. Cell culture and subculture

[0060] 1. The super-clean bench is sterilized for 30 min, and after sterilization, it is ventilated for 10 min.

[0061] 2. Wear protective articles (mask, gloves, protective clothing).

[0062] 3. Observe the cells, and when the cell confluence reaches 90%, the cells can be subcultured.

[0063] 4. After removing the supernatant, wash the cells with PBS for 1-2 times.

[0064] 5. Add an appropriate amount of trypsin to digest the cells for about 8 min.

[0065] 6. Observe whether the cells are detached, and then add complete culture medium to stop digestion.

[0066] 7. At this time, the cells can be subcultured at 1:3-1:6.

[0067] 8. After subculture, the cells are placed in a 5% CO2, 37°C incubator for culture.

[0068] II. Cell plating

[0069] 1. Observe the cell state and confluence rate before cell plating.

[0070] 2. Centrifuge after cell digestion.​

[0071] 3. Stain cells with trypan blue and count cells under microscope.

[0072] 5. Cell plating density 1*10^6 cells / well, 2 mL / well.

[0073] 6. Plated cells are incubated in the incubator for 24 h.

[0074] III. Add test sample

[0075] 1. Add active at appropriate concentration and incubate for 72 h.

[0076] 2. Remove supernatant.

[0077] 3. Wash twice with PBS.

[0078] 4. Digest cells with trypsin.

[0079] 5. Centrifuge to remove supernatant.

[0080] 6. Wash once with PBS.

[0081] 7. Prepare 1% triton-100 in PBS and add 200 μL.

[0082] 8. Freeze sample at -80°C for 30 min.

[0083] 9. Incubate at room temperature for 10 min.

[0084] 10. Centrifuge at 12,000 rpm for 5 min.

[0085] 11. Test: add 90 μL supernatant and 10 μL of 5 mg / mL dopamine solution.

[0086] 12. Incubate at 37°C for 1-2 h (2 h for this experiment).

[0087] 13. Read at 475 nm.

[0088] Results of the experiment are shown in the accompanying Figure 4 .

[0089] Figure 4 A comparison of the results of the cell melanin inhibition test of the high-permeability-through-skin arbutin analog obtained according to the preparation method of Example 1 of the present application and arbutin of Comparative Example 1 is shown in the accompanying Figure 4 As can be seen, the arbutin analog of Example 1 (Compound III) and arbutin of Comparative Example 1 have little difference in melanin inhibition effect, and as the sample concentration increases, the melanin inhibition effect of the arbutin analog of Example 1 (Compound III) is significantly stronger than that of arbutin of Comparative Example 1.

[0090] The above merely describes preferred embodiments of the present application, and is not intended to limit the present application in any form. Although the present application has been disclosed with the preferred embodiments as above, it is not intended to limit the present application. Any person skilled in the art can make some changes or modifications to the above disclosed methods and technical contents to make equivalent embodiments with equivalent changes, without departing from the technical solution of the present application. Any simple modification, equivalent change and modification made to the above embodiments according to the technical essence of the present application, without departing from the technical solution of the present application, still belongs to the scope of the technical solution of the present application.

Claims

1. A method for preparing a highly efficient transdermal arbutin analogue, characterized in that, This method includes the following steps: Compound I and Compound II undergo esterification in the presence of a solvent and a catalyst to obtain Compound III. The synthetic route is as follows: ； in, In compound II, X represents one of F, Cl, Br, and I; R represents a C1-C30 alkyl group. The esterification reaction temperature is 60~80℃; The esterification reaction time is 3-8 hours; The catalyst is selected from one or a mixture of several of pyridine, triethylamine, diethylamine, isopropylamine, diisopropylethylamine, N-methylmorpholine, and tetramethylethylenediamine.

2. The preparation method according to claim 1, characterized in that, The solvent is selected from one or a mixture of several of dimethyl sulfoxide, N,N-dimethylformamide, acetonitrile, dichloromethane, chloroform, and carbon tetrachloride.

3. The preparation method according to claim 1, characterized in that, The solvent is N,N-dimethylformamide.

4. The preparation method according to claim 1, characterized in that, The catalyst is pyridine.

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