Compounds capable of inhibiting heparanase and mmp activities simultaneously and uses thereof

Abstract

The present application provides a compound capable of simultaneously inhibiting heparanase and MMP activity, which is a compound shown in the following general formula: wherein A1 is a carbon atom or a nitrogen atom, A2 is a carbon atom; the bond A1-A2 is a single bond, a double bond, or is connected with a benzene ring or a cyclohexene ring; the five-membered ring can also be replaced by a six-membered ring. The compound has the functions of maintaining the integrity of epidermal structure, promoting the differentiation of keratinocytes, and regulating the function of skin barrier.

Description

Technical Field

[0001] This invention relates to the field of new cosmetic materials, specifically to compounds capable of simultaneously inhibiting the activity of heparinase and MMP, and their applications. Background Technology

[0002] The skin is a vital barrier protecting the body from external environmental factors. The epidermal basement membrane (BM), located at the junction of the epidermis and dermis, plays a crucial role in maintaining the integrity of the epidermal structure, promoting keratinocyte differentiation, and regulating skin barrier function. In daily life, facial skin is frequently exposed to sunlight, especially ultraviolet (UV) radiation, which can damage the structure and function of the epidermal basement membrane. This can lead to decreased skin barrier function, increased transepidermal water loss, and problems such as dryness and roughness.

[0003] Studies have shown that ultraviolet (UV) radiation can induce the activation of various basement membrane degrading enzymes, including heparanase and matrix metalloproteinases (MMPs). These enzymes can degrade major components of the basement membrane, such as heparan sulfate proteoglycans and type IV collagen, thereby disrupting the connection between the epidermis and dermis and weakening the skin's barrier function and repair capabilities. Therefore, effectively inhibiting the activity of these basement membrane degrading enzymes and protecting skin structure and barrier function has become an important research topic in dermatology and cosmetics.

[0004] In existing technologies, although some antioxidants, metal chelators, or enzyme inhibitors have been used to slow down skin aging caused by ultraviolet radiation, most of them have a single mechanism of action or only target a certain type of metalloproteinase, making it difficult to effectively inhibit the activity of multiple basement membrane degrading enzymes at the same time, resulting in limited protective effects. Summary of the Invention

[0005] The present invention aims to overcome the above-mentioned defects and develop a compound that can simultaneously inhibit the activity of heparinase and multiple MMPs to prevent or improve sun damage to the epidermal basement membrane, thereby enhancing the skin barrier function. It has important research value and application prospects.

[0006] This invention provides a compound capable of simultaneously inhibiting the activities of heparinase and MMP, which is a compound represented by the following general formula: ; Among them, A1 and A2 are carbon atoms; Bonds A1-A2 can be single or double bonds, or be bonded to a benzene ring, or to a cyclohexene ring; Furthermore, the present invention provides a compound that can simultaneously inhibit the activity of heparinase and MMP, and also has the characteristic that the five-membered ring can be replaced by a six-membered ring.

[0007] Furthermore, the present invention provides a compound capable of simultaneously inhibiting the activity of heparinase and MMP, and also has the characteristic that at least one of the carbonyl groups is reduced to -CH2- or hydroxyl, or the reduced hydroxyl group is etherified to form an alkoxy group.

[0008] Furthermore, the present invention provides a compound capable of simultaneously inhibiting heparinase and MMP activity, and also has the characteristic that it is a compound with the following structure: .

[0009] Furthermore, this invention also suggests the application of the above-mentioned compounds in the preparation of cosmetics.

[0010] Furthermore, this invention also suggests the application of the above-mentioned compounds in the preparation of anti-aging products.

[0011] Furthermore, the present invention also suggests the use of the above-mentioned compounds as inhibitors of heparanase and matrix metalloproteinase.

[0012] In addition, the present invention also provides a method for preparing the above-mentioned compound, specifically as follows: The compounds shown in formulas 6, 7, and 10, in an alcohol solvent, are derived from N-(2-hydroxyethyl)diamid derivatives via a reducing agent to produce the target product. Notably, the compound of formula 7 produces a product with a methoxy group on one side. Attached Figure Description

[0013] Figure 1 Expression results of MMP1 in different molecules.

[0014] Figure 2 Expression results of MMP2 in different molecules.

[0015] Figure 3 Expression results of MMP9 in different molecules.

[0016] Figure 4 The expression results of human heparin of different molecules.

[0017] Figure 5 Statistical results for molecule 1.

[0018] Figure 6 Statistical results for molecule 2.

[0019] Figure 7 Statistical results for molecule 3.

[0020] Figure 8 Statistical results for molecule 4.

[0021] Figure 9Statistical results for molecule 5.

[0022] Figure 10 Statistical results for molecule 6.

[0023] Figure 11 Statistical results for molecule 7.

[0024] Figure 12 Statistical results for molecule 8.

[0025] Figure 13 Statistical results for molecule 9.

[0026] Figure 14 Statistical results for molecule 10.

[0027] Figure 15 Collagen IV fluorescence microscopy images of molecules 1 and 6.

[0028] Figure 16 Collagen IV statistics for molecules 1 and 6.

[0029] Figure 17 Collagen VII fluorescence microscopy results of molecules 1 and 6.

[0030] Figure 18 Collagen VII statistics for molecules 1 and 6. Detailed Implementation

[0031] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0032] This embodiment provides a compound that can simultaneously inhibit the activities of heparinase and MMP, and is a compound represented by the following general formula: ; Among them, A1 and A2 are carbon atoms; Bonds A1-A2 can be single or double bonds, or be bonded to a benzene ring, or to a cyclohexene ring; In some cases, at least one of the carbonyl groups is reduced to -CH2- or a hydroxyl group, or the reduced hydroxyl group is etherified to form an alkoxy group.

[0033] In some cases, a five-membered ring can also be replaced by a six-membered ring; The preferred compounds are as follows: .

[0034] Among them, the CAS number of Equation 2 (Molecule 2) is 3445-11-2, the CAS number of Equation 3 (Molecule 3) is 18190-44-8, the CAS number of Equation 4 (Molecule 4) is 3891-07-4, the CAS number of Equation 5 (Molecule 5) is 1585-90-6, the CAS number of Equation 8 (Molecule 8) is 38772-50-8, and the CAS number of Equation 9 (Molecule 9) is 3445-12-3. The compounds represented by formulas 6 (Molecule 6), 7 (Molecule 7), and 10 (Molecule 10) are derived from N-(2-hydroxyethyl)diamid derivatives in an alcohol solvent via the action of a reducing agent to obtain the target product. The molar ratio of N-(2-hydroxyethyl)diamid derivative to reducing agent is 1:2-10.

[0035] The specific synthesis methods are shown in Examples 1-3 below.

[0036] Example 1. Synthesis method of Molecule6

[0037] In an alcohol solvent, N-(2-hydroxyethyl)diamide derivative N-(2-hydroxyethyl)butadiamide and NaBH were mixed in a molar ratio of 1:5-10 and reacted at room temperature for 20+ hours. After acid quenching, the liquid phase was collected and purified to obtain the target product.

[0038] The specific synthesis process is as follows: In a 500ml three-necked flask, 14.3g (0.1mol) of N-(2-hydroxyethyl)butadiamide and 115g of methanol were added and dissolved under magnetic stirring. The mixture was cooled to 0-10℃ in an ice-water bath, and 0.2g (0.8mol) of NaBH4 was added in portions. The temperature was then gradually increased to 20-30℃ and reacted for 24h. After the reaction was completed, the temperature was lowered to below 20℃, and the pH of the reaction solution was quenched to below 3 with 150g of 6M hydrochloric acid. The insoluble matter was removed by filtration, and the filtrate was evaporated until no liquid remained. 150g of acetone was added to dissolve the insoluble matter, and the insoluble matter was filtered. After evaporating the acetone to dryness, column chromatography was performed to obtain 8.1g of product, 1-(2-hydroxyethyl)pyrrolidine-2,5-diol, with a yield of 55% and a purity of 98%.

[0039] 1 H NMR (400 MHz, DMSO- d6) δ 4.67 – 4.58 (m, 4H), 4.43 – 4.30 (m, 2H), 3.82 – 3.64 (m, 3H), 3.12 – 2.96 (m, 3H), 1.87 – 1.65 (m, 5H).

[0040] 13 C NMR (100 MHz, DMSO-d6) δ 82.76, 59.06, 45.73, 29.05.

[0041] M / Z:147.18.

[0042] Example 2. Synthesis method of Molecule7

[0043] In methanol solvent, N-(2-hydroxyethyl)diamide derivative N-hydroxyethyl phthalamide and NaBH were mixed in a molar ratio of 1:2-6 and reacted at room temperature for 2+ hours. After acid quenching, the liquid phase was collected and purified to obtain the target product.

[0044] Specific synthesis process: In a 500ml three-necked flask, 19.1g (0.1mol) of N-hydroxyethyl phthalamide and 153g of methanol were added and dissolved under magnetic stirring. The mixture was cooled to 0-10℃ in an ice-water bath. 15.1g (4eq) of NaBH4 was added in portions, and the temperature was gradually increased to 20-30℃ for 5 hours. After the reaction was completed, the temperature was lowered to below 20℃, and the pH of the reaction solution was quenched to below 3 with 90g of 6M hydrochloric acid. The insoluble matter was removed by filtration. The filtrate was evaporated until no liquid remained. 200g of ethyl acetate and 200mL of water were added, and the ethyl acetate layer was collected separately. After evaporating the ethyl acetate to dryness, column chromatography was used to purify the product 13.0g, 2-(2-hydroxyethyl)-3-methoxy-2,3-dihydro-1 H -Isoindol-1-one, yield 63%, purity 98%.

[0045] 1 H NMR (400 MHz, DMSO- d 6) δ 7.77 (dd, J = 7.2, 1.8 Hz, 1H), 7.64 (ddd, J = 7.3, 1.7, 0.7 Hz, 1H), 7.50 (dtd, J = 18.7, 7.5, 1.7 Hz, 2H), 6.15 (tq, J = 1.4, 0.6 Hz, 1H), 4.66 (t,J = 7.4 Hz, 1H), 3.93 – 3.70 (m, 4H), 3.34 (d, J = 1.4 Hz, 3H).

[0046] 13 C NMR (100 MHz, DMSO- d 6) δ 166.15, 140.93, 132.01, 130.73, 126.21, 126.10, 124.67, 90.72, 59.78, 54.00, 45.07.

[0047] M / Z:207.25.

[0048] Example 3. Synthesis method of Molecule10

[0049] In an alcohol solvent, N-(2-hydroxyethyl)diamide derivative N-(2-hydroxyethyl)-1,2,3,6-tetrahydrophthalimide was mixed with NaBH at a molar ratio of 1:4-8 and reacted at room temperature for 10+ hours. After acid quenching, the liquid phase was collected and purified to obtain the target product.

[0050] Specific synthesis process: In a 500ml three-necked flask, 19.5g (0.1mol) of N-(2-hydroxyethyl)-1,2,3,6-tetrahydrophthalimide and 156g of methanol were added and dissolved under magnetic stirring. The mixture was cooled to 0-10℃ in an ice-water bath, and 22.6g (6eq) of NaBH4 were added in portions. The temperature was then gradually increased to 20-30℃ and the reaction was carried out for 15 hours. After the reaction was completed, the temperature was lowered to below 20℃, and the pH of the reaction solution was quenched to below 3 with 120g of 6M hydrochloric acid. The insoluble matter was removed by filtration, and the filtrate was evaporated until no liquid remained. The product was purified by column chromatography to obtain 14.1g of product (3a). R 7a S )-2-(2-hydroxyethyl)-2,3,3a,4,7,7a-hexahydro-1 H -Isoindole-1,3-diol, yield 70.7%, purity 97%.

[0051] 1 H NMR (400 MHz, DMSO- d 6) δ 5.67 – 5.55 (m, 2H), 4.69 (d, J = 5.2 Hz, 2H), 4.60 (t, J = 7.5 Hz, 1H), 4.38 (dd,J = 7.1, 5.3 Hz, 2H), 3.68 (q, J =7.1 Hz, 2H), 3.11 – 2.96 (m, 2H), 2.26 – 2.14 (m, 2H), 2.10 – 1.98 (m, 2H), 1.97 – 1.85 (m, 2H).

[0052] 13 C NMR (100 MHz, DMSO- d 6) δ 128.68, 85.85, 59.53, 47.10, 41.40, 40.92, 27.86.

[0053] M / Z:199.27.

[0054] Example 4. Performance Test A (MMP1, 2, 9, and human heparin) 1. Materials and Methods 1.1 Cell Culture A431 cells (Shanghai Enzyme-Link Biotechnology Co., Ltd., catalog number: ml082207) were seeded in culture plates and cultured until they reached the logarithmic growth phase. Ten small molecules were added at concentrations of 0.3%, 0.5%, and 1% for incubation, and the cells were examined at 6, 12, and 24 hours. 1.2 Analysis of matrix metalloproteinase MMP-1 According to the reagent operation instructions, the expression of MMP-1 was detected using a human matrix metalloproteinase 1 (MMP-1) enzyme-linked immunosorbent assay kit (catalog number: CSB-E04672h, CUSABIO, China). The control group was treated with DMSO.

[0055] 1.3 Analysis of matrix metalloproteinase MMP-2 According to the reagent instructions, the expression of MMP-2 was detected using the human matrix metalloproteinase 2 / gelatinase A (MMP-2 / Gelatinase A) enzyme-linked immunosorbent assay kit (catalog number: CSB-E04675h, CUSABIO, China). The control group was treated with DMSO.

[0056] 1.4 Analysis of matrix metalloproteinase MMP-9 According to the reagent operation instructions, the expression of MMP-9 was detected using the human matrix metalloproteinase 9 / gelatinase B (MMP-9 / Gelatinase B) enzyme-linked immunosorbent assay kit (catalog number: CSB-E08006h, CUSABIO, China). The control group was treated with DMSO.

[0057] 1.5 Molecular analysis of human heparin According to the reagent operation instructions, the expression of heparin molecules was detected using the Human Low Molecular Weight Heparin (LMWH) ELISA Kit (catalog number: ARD11433, Guangzhou Orida Biotechnology Co., Ltd., China), and the control group was treated with DMSO.

[0058] 1.6. Known reference point for comparison Molecule 1 CAS: 3699-54-5.

[0059] 2 Results 2.1 Expression trends of matrix metalloproteinases and heparin molecules 2.1.1 MMP-1 Expression Trends like Figure 1 As shown, the overall MMP1 expression values ​​of the 10 small molecules ranged from 0.4 ng / ml to 1.0 ng / ml.

[0060] Among them, Molecule 3, 4, 5, 6, and 9 showed high MMP1 expression levels, exceeding 0.8 ng / ml. The remaining small molecule MMP1 expression levels were moderate.

[0061] 2.1.2 MMP-2 Expression Trends like Figure 2 As shown, the MMP2 expression values ​​of the 10 small molecules generally ranged from 0 ng / ml to 2.7 ng / ml.

[0062] Molecules 3, 4, 5, and 9 are molecules that highly express MMP2. Molecules 1 and 2 are molecules that lowly express MMP2.

[0063] 2.1.3 MMP-9 Expression Trends like Figure 3 As shown, the overall MMP1 expression values ​​of the 10 small molecules ranged from 0 ng / ml to 0.35 ng / ml.

[0064] Molecules 3, 4, and 9 were highly expressed. Molecules 1, 2, 5, and 6 were consistently expressed at the lowest levels, essentially not expressed at all.

[0065] 2.1.4 Trends in Heparin Molecular Expression like Figure 4 As shown, the expression levels of Heparin for the 10 small molecules ranged from 40 ng / ml to 130 ng / ml.

[0066] Among them, Molecule 1, 5, 7, 9, and 10 showed high Heparin expression levels, exceeding 80 ng / ml. The remaining small molecule Heparin showed moderate expression levels.

[0067] 2.2 Effects of different small molecules on matrix metalloproteinases and heparin molecules 2.2.1 Small molecule No. 1 like Figure 5 As shown, MMP1 showed concentration-dependent differences, with statistically significant differences between 0.3% and 0.5%, and between 0.3% and 1%, with 0.3% enhancing its expression level.

[0068] MMP2: There was a time-dependent difference, with a statistically significant difference between 12h and 24h.

[0069] MMP9: Completely inhibit its expression level.

[0070] Heparin: There was a concentration-dependent difference; a statistically significant difference was observed between 0.3% and 1%, with 0.3% enhancing its expression level. Figure 5 ) 2.2.2 Small molecule No. 2 like Figure 6 As shown, MMP1 exhibits time-dependent differences, with statistically significant differences between 6h and 12h, and between 6h and 24h.

[0071] MMP2: There were time-dependent differences, with statistically significant differences between 12h and 24h, and between 6h and 12h.

[0072] MMP9: Completely inhibit its expression level. Heparin: There are time-dependent differences, with statistically significant differences between 12h and 24h, and between 6h and 24h, with an increase in expression levels at 24h.

[0073] 2.2.3 Small molecule No. 3 like Figure 7 As shown, MMP1 showed concentration-dependent differences, with statistically significant differences between 0.3% and 0.5%, and between 0.3% and 1%, indicating high overall expression levels.

[0074] MMP2: There are time-dependent differences, with statistically significant differences between 12h and 6h, and between 12h and 24h. Overall expression levels are high.

[0075] MMP9: No statistically significant differences were found in concentration and time.

[0076] Heparin: showed concentration- and time-dependent differences, with statistically significant differences between 6h and 24h, and between 0.3% and 1%. Overall expression levels decreased over time.

[0077] 2.2.4 Small molecule number 4 like Figure 8 As shown, MMP1 showed concentration-dependent differences, with a statistically significant difference between 0.3% and 1%, and the overall expression levels were both high.

[0078] MMP2: There were concentration-dependent differences, with statistically significant differences between 0.3% and 1%, and between 0.5% and 1%, and the overall expression levels were high.

[0079] MMP9: No statistically significant differences were found in concentration and time.

[0080] Heparin: There are time-dependent differences, with statistically significant differences between 12h and 6h, and between 24h and 6h. Overall expression levels are moderate.

[0081] 2.2.5 Small molecule No. 5 like Figure 9 As shown, MMP1 exhibited concentration- and time-dependent differences, with statistically significant differences between 12h and 24h, 6h and 24h, and 0.3% and 1%, respectively, and overall expression levels were high.

[0082] MMP2: There are concentration- and time-dependent differences, with statistical differences between 6h and 24h, and between 0.3% and 1%. High concentrations over a long period of time increase its expression level.

[0083] MMP9: Completely inhibit its expression level.

[0084] Heparin: showed concentration-dependent differences, with statistically significant differences between 0.3% and 0.5%, and between 0.3% and 1%, with overall expression levels being high.

[0085] 2.2.6 Small molecule number 6 like Figure 10 As shown, MMP1 exhibits time-dependent differences, with statistically significant differences between 12h and 24h, and between 6h and 24h, and overall expression levels are high.

[0086] MMP2: There were statistically significant time-dependent differences between 12h and 24h, and between 6h and 24h, with overall expression levels being low.

[0087] MMP9: Completely inhibit its expression level.

[0088] Heparin: No statistically significant differences were observed in concentration and time.

[0089] 2.2.7 Small molecule No. 7 like Figure 11 As shown, MMP1 exhibits time-dependent differences, with statistically significant differences between 12h and 24h, and between 6h and 24h, and its overall expression level is moderate.

[0090] MMP2: There are time-dependent differences, with statistically significant differences between 12h and 24h, and between 6h and 24h, and the overall expression level is moderate.

[0091] MMP9: There are time-dependent differences, with statistical differences between 6h and 24h, and the overall expression level is moderate.

[0092] Heparin: showed concentration-dependent differences, with statistically significant differences between 0.3% and 0.5%, and between 0.3% and 1%, with overall expression levels being high.

[0093] 2.2.8 Small molecule No. 8 like Figure 12 As shown, MMP1 exhibits time-dependent differences, with statistically significant differences between 12h and 24h, and between 6h and 24h, and its overall expression level is moderate.

[0094] MMP2: There were time-dependent differences, with statistically significant differences between 12h and 24h, 6h and 24h, and 6h and 12h, and the overall expression level was low.

[0095] MMP9: There are time-dependent differences, with statistically significant differences between 6h and 12h, and between 6h and 24h, and the overall expression level is moderate.

[0096] Heparin: showed time-dependent differences, with statistically significant differences between 6h and 24h, and overall expression levels were moderate.

[0097] 2.2.9 Small molecule number 9 like Figure 13 As shown, MMP1 showed concentration-dependent differences, with statistically significant differences between 0.5% and 1%, and between 0.3% and 1%, with overall expression levels being high.

[0098] MMP2: There were concentration-dependent differences, with statistically significant differences between 0.5% and 0.3%, and between 0.3% and 1%, and the overall expression levels were high.

[0099] MMP9: There were concentration-dependent differences, with statistical differences between 0.5% and 0.3%, and between 0.5% and 1%. At 0.3%, it was expressed at all time points.

[0100] Heparin: showed concentration-dependent differences, with statistically significant differences between 0.3% and 1%, and between 0.5% and 1%, with the highest expression level at 0.5%.

[0101] 2.2.10 Small molecule No. 10 like Figure 14 As shown, MMP1 showed concentration-dependent differences, with statistically significant differences between 0.5% and 1%, and between 0.3% and 1%, with overall expression levels being high.

[0102] MMP2: There were concentration- and time-dependent differences, with statistically significant differences between 6h and 24h, and between 0.3% and 1%, with overall expression levels being low.

[0103] MMP9: There are concentration-dependent differences, with statistical differences between 0.5% and 1%, and between 0.3% and 1%. At 1%, it is expressed at all time points.

[0104] Heparin: showed concentration-dependent differences, with statistically significant differences between 0.3% and 1%, 0.5% and 1%, and 0.5% and 0.3%. Overall expression levels were high, with the highest expression level at 1%.

[0105] Example 5. Performance Test B (Collagen IV, Collagen VII, Transepidermal Water Loss (TEWL) Test) 5.1. Test methods: The tests were conducted according to the "Detection Method of Type IV Collagen and Type VII Collagen Content Based on UVA Irradiation of Fibroblasts" and the "Detection Method of Transepidermal Water Loss (TEWL) Based on Barrier-Weakened 3D Epidermal Skin Model (EpiKutis®)".

[0106] 5.2. Grouping Blank control group (BC) Negative control group (NC) Positive control group (PC), TGF-β1, administered at a concentration of 100 ng / mL Sample group 1 molecule, administered at concentrations of 0.01% (m / m), 0.05% (m / m), and 0.1% (m / m). Sample group 6 molecule, administered at concentrations of 0.01% (m / m), 0.05% (m / m), and 0.1% (m / m).

[0107] 5.3. Results 5.3.1. Collagen IV like Figure 15-16 As shown in Table 1 below, the Collagen IV content in the NC group was significantly lower than that in the BC group, indicating that the test stimulus conditions were effective.

[0108] Compared with the NC group, the Collagen IV content in the PC group was significantly increased, indicating that the positive control in this test was effective.

[0109] Compared with the NC group, the Collagen IV content of samples with molecules 1-0.01%, 1-0.05%, 1-0.1%, 6-0.05%, and 6-0.1% increased significantly, with increases of 18.60%, 67.44%, 76.74%, 34.88%, and 83.72%, respectively. This indicates that molecules 1 and 6 can effectively promote the production of type IV collagen.

[0110] Table 1. Statistical results of Collagen IV Group Relative IOD / average cell count SD -value Lift Rate (NC) BC 1 0.08 / / NC 0.43 0.03 0.000 ## / PC 0.78 0.05 0.000 ** 81.40% Molecular weight 1-0.01% 0.51 0.02 0.020 * 18.60% Molecular weight 1-0.05% 0.72 0.05 0.001 ** 67.44% Molecular weight 1-0.1% 0.76 0.01 0.000 ** 76.74% Molecular weight 6-0.01% 0.48 0.02 0.089 / Molecular weight 6-0.05% 0.58 0.01 0.001 ** 34.88% Molecular weight 6-0.1% 0.79 0.08 0.002 ** 83.72% Note: When performing statistical analysis using the t-test method, significance compared with the BC group is indicated by #, with P-value < 0.05 indicated by # and P-value < 0.01 indicated by ##; significance compared with the NC group is indicated by *, with P-value < 0.05 indicated by * and P-value < 0.01 indicated by **.

[0111] 5.3.2. Collagen VII like Figure 17-18 As shown in Table 2 below, the Collagen VII content in the NC group was significantly lower than that in the BC group, indicating that the stimulation conditions in this test were effective.

[0112] Compared with the NC group, the Collagen VII content in the PC group was significantly increased, indicating that the positive control in this test was effective.

[0113] Compared with the NC group, the Collagen VII content of samples with molecules 1-0.01%, 1-0.05%, 1-0.1%, 6-0.01%, 6-0.05%, and 6-0.1% increased significantly, with increases of 42.86%, 97.14%, 145.71%, 31.43%, 108.57%, and 171.43%, respectively. This indicates that molecules 1 and 6 can effectively promote the production of type IV collagen.

[0114] Table 2. Statistical results of Collagen VII Group Relative IOD / average cell count SD -value Lift Rate (NC) BC 1 0.07 / / NC 0.35 0.03 0.000 ## / PC 0.68 0.02 0.000 ** 94.29% Molecular weight 1-0.01% 0.5 0.08 0.038 * 42.86% Molecular weight 1-0.05% 0.69 0.15 0.017 * 97.14% Molecular weight 1-0.1% 0.86 0.1 0.001 ** 145.71% Molecular weight 6-0.01% 0.46 0.04 0.021 * 31.43% Molecular weight 6-0.05% 0.73 0.06 0.001 ** 108.57% Molecular weight 6-0.1% 0.95 0.02 0.000 ** 171.43% Note: When performing statistical analysis using the t-test method, significance compared with the BC group is indicated by #, with P-value < 0.05 indicated by # and P-value < 0.01 indicated by ##; significance compared with the NC group is indicated by *, with P-value < 0.05 indicated by * and P-value < 0.01 indicated by **.

[0115] 5.3.3. Transepidermal water loss (TEWL) The results are shown in Table 3. Compared with the BC group, the TEWL value of the PC group did not change significantly.

[0116] Compared with the BC group, the TEWL values ​​of samples with molecule 1-0.3% and molecule 6-0.3% decreased significantly, with inhibition rates of 33.73% and 32.87%, respectively. This means that molecules 1 and 6 can effectively alleviate epidermal water loss.

[0117] Table 3. Summary of Transepidermal Water Loss (TEWL) Test Results Group Average value (g / m2·h) SD -value Inhibition rate (BC) BC 20.87 2.32 / / PC 21.04 4.7 0.958 / Molecular weight 1-0.3% 13.83 0.18 0.006** 33.73% Molecular weight 6-0.3% 14.01 0.83 0.008** 32.87% Note: Use t When performing statistical analysis using the -test method, significance compared to the BC group is indicated by *. P -value < 0.05 is represented as *. P -value < 0.01 It is represented as **.

[0118] In summary, based on UVA irradiation of fibroblasts, compared with the control group, sample molecule 6 showed a significant increase in type VII collagen content at a concentration of 0.01% (m / m), with an increase rate of 31.43%, indicating that the sample at this concentration can increase type VII collagen content and has anti-wrinkle effects. At a concentration of 0.05% (m / m), the contents of both type IV and type VII collagen increased significantly, with increase rates of 34.88% and 108.57%, respectively, indicating that the sample at this concentration can increase the contents of type IV and type VII collagen and has firming and anti-wrinkle effects. At a concentration of 0.1% (m / m), the content of type IV collagen and type VII collagen increased significantly, with increases of 83.72% and 171.43%, respectively. This indicates that the sample can increase the content of type IV collagen and type VII collagen at this concentration, and has firming and anti-wrinkle effects.

[0119] Based on the 3D epidermal skin model (EpiKutis®), compared with the control group, the transepidermal water loss (TEWL) of sample molecule 6 at a concentration of 0.3% (m / m) was significantly reduced, with an inhibition rate of 32.87%, indicating that the sample can downregulate transepidermal water loss (TEWL) at this concentration and has a repairing effect.

Claims

1. A compound capable of simultaneously inhibiting the activities of heparinase and MMP, characterized in that: Compounds represented by the following general formula: ； Among them, A1 and A2 are carbon atoms; Bonds A1-A2 can be single or double bonds, or can be bonded to a benzene ring or a cyclohexene ring.

2. The compound as described in claim 1, capable of simultaneously inhibiting heparinase and MMP activity, characterized in that: At least one of the carbonyl groups is reduced to -CH2- or hydroxyl.

3. The compound as described in claim 2, capable of simultaneously inhibiting heparinase and MMP activity, characterized in that: The reduced hydroxyl group is then etherified to form an alkoxy group.

4. The compound as described in claim 2, capable of simultaneously inhibiting heparinase and MMP activity, characterized in that: The five-membered ring is replaced by a six-membered ring.

5. A compound capable of simultaneously inhibiting the activity of heparinase and MMP, characterized in that... The compound has the following structure: 。 6. The use of the compound as described in any one of claims 1-4, capable of simultaneously inhibiting heparinase and MMP activity, in the preparation of cosmetics.

7. The use of the compound as described in any one of claims 1-4, which can simultaneously inhibit the activity of heparinase and MMP, in the preparation of anti-aging products.

8. The use of the compound as described in any one of claims 1-4, which can simultaneously inhibit the activity of heparinase and MMP, in the preparation of products that maintain the integrity of the epidermal structure, promote keratinocyte differentiation, and regulate the skin barrier function.

9. The use of the compound as described in any one of claims 1-4, which is capable of simultaneously inhibiting the activity of heparinase and MMP, as an inhibitor of heparanase and matrix metalloproteinase.

10. A method for preparing a compound capable of simultaneously inhibiting heparinase and MMP activity as described in any one of claims 1-4, characterized in that: In an alcohol solvent, the target product is obtained by reacting an N-(2-hydroxyethyl)diamide derivative as a raw material with a reducing agent.

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