Water-soluble vitamin e derivative, method of preparation and use thereof

Abstract

The present application relates to the technical field of compound preparation, in particular to a water-soluble vitamin E derivative, a preparation method and application thereof, and the parent nucleus is shown in the following formula: wherein R is cycloalkyl or heterocyclic group, the compound has good anti-inflammatory and antioxidant functions, realizes anti-inflammatory and anti-aging through the pathways of inhibiting inflammatory factor expression, promoting collagen production, promoting self-repair of skin barrier, etc., and has good application prospect.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of compound preparation, in particular to a water-soluble vitamin E derivative, a preparation method and application thereof. BACKGROUND

[0002] Natural products are often used as the starting point for the design of new drugs, and on this basis, structural modification and modification are carried out, so that the compound has new activity and function. The structural modification of natural product alpha-tocopherol (vitamin E) shows that the long-chain alkyl group on the side chain is replaced by a carboxyl group to obtain water-soluble vitamin E, which is called Trolox. Trolox is reported to have a wide range of biological activities, including anticancer, neuroprotection, antioxidant and the like. Trolox is often used as a positive control to measure the antioxidant capacity of other antioxidants. Trolox can increase the wound healing ability by reducing oxidative stress; through the mechanism of scavenging reactive oxygen species (ROS), it has a protective effect on lipopolysaccharide-induced oxidative stress and inflammation, and is a potential antioxidant against cell oxidative damage.

[0003] When the skin is exposed to low doses of UVB radiation, ROS is produced in the cells, and intracellular enzymatic antioxidants such as superoxide dismutase (SOD) and catalase (CAT) and non-enzymatic antioxidants such as reduced glutathione (GSH) are activated to counteract or neutralize ROS, thereby maintaining the skin redox balance and reducing cellular oxidative toxicity. However, excessive UVB radiation can cause excessive accumulation of ROS, disrupting the balance between the oxidation and antioxidant systems, and causing severe oxidative stress. In addition, ROS-mediated oxidative stress can cause oxidative damage to various biological macromolecules such as proteins, DNA, and lipids. This damage causes peroxidation of cell membrane lipids, promotes the release of lactate dehydrogenase (LDH), reduces the levels of SOD, CAT, and GSH, and increases the levels of lipid peroxide (LPO) and malondialdehyde (MDA). The p38 MAPK signaling pathway is specifically activated by phosphorylation under the stress of UVB radiation and oxidative damage, and the p38 MAPK signaling pathway is generally involved in the process of photodamage and oxidative damage. Generally, for UVB radiation, the skin can initiate a series of self-repair functions such as DNA repair, epidermal thickening, and the use of antioxidant factors to counteract photodamage, but for severe photodamage, this self-repair is limited. A single excessive exposure to UVB radiation can cause various skin changes, including erythema, sunburn, vascular hyperpermeability, edema, pain, and fever, which are characteristic of acute photodamage and are often closely related to oxidative stress. Therefore, antioxidants play an important role in the repair process of acute photodamage, and general antioxidant methods include applying sunscreen and using various antioxidants. However, most sunscreens with high sun protection factor contain metal oxide nanoparticles and form various organic ingredients that absorb ultraviolet rays, and the use of these sunscreens is limited because some of the ingredients are toxic and affect the environment and health. The most widely used antioxidants, such as vitamin C (Vc) and vitamin E (Ve), also have defects, for example, the use of Vc at high doses can increase the body's sensitivity to harmful substances and cause cell damage. Ve can cause nausea, headache, gastrointestinal discomfort, and the risk of hemorrhagic stroke. Therefore, it is necessary and necessary to develop new natural antioxidants against photodamage.

[0004] In view of the above defects, the inventor of the present application has finally obtained the present application after a long period of research and practice. SUMMARY

[0005] The purpose of the present application is to solve the problem of how to prepare a new natural antioxidant against photodamage, and to provide a water-soluble vitamin E derivative, a preparation method and applications thereof.

[0006] In order to achieve the above-mentioned purpose, the application discloses a water-soluble vitamin E derivative, which is a 6-hydroxybenzopyran derivative, and the parent nucleus of the 6-hydroxybenzopyran derivative is shown in the following formula:

[0007]

[0008] In the formula, R is a cycloalkyl group or a heterocyclic group.

[0009] The water-soluble vitamin E derivative is any one of the following:

[0010]

[0011] The application further discloses a preparation method of the water-soluble vitamin E derivative, and the preparation method is as follows: an alcohol compound and the water-soluble vitamin E are subjected to a substitution reaction under alkaline conditions to obtain the 6-hydroxybenzopyran derivative.

[0012] The reaction process is shown in the following formula:

[0013]

[0014] In the formula, R is a cycloalkyl group or a heterocyclic group.

[0015] The alcohol compound is any one of cyclopropyl alcohol, cyclopropylmethanol, (S)-glycidol, (R)-glycidol, cyclobutanol, 3-hetero oxygen cyclobutanol, cyclopentanol, cyclohexanol, 4-hydroxyfuran, 2-hydroxyfuran, 4-hydroxycyclohexanone and 1-methyl-4-piperidinol.

[0016] The application further discloses application of the water-soluble vitamin E derivative in preparation of anti-inflammatory and antioxidant products.

[0017] Compared with the prior art, the application has the following beneficial effects:

[0018] 1. The preparation process is simple, the recovery rate is high, and the operation is convenient.

[0019] 2. The water-soluble vitamin E derivative prepared by the application is an alicyclic ester or a heterocyclic fat, retains the antioxidant activity of Trolox, enhances the anti-inflammatory activity, has balanced antioxidant and anti-inflammatory activity, and has better oral bioavailability.

[0020] 3. The water-soluble vitamin E derivative prepared by the application is an effective component for relieving inflammation, tissue repair and ultraviolet damage protection and treatment, and the compound has good anti-inflammatory, antioxidant and repair activities, and can be used for improving skin condition, whitening, sun protection, anti-aging and the like, and has good application prospect and potential. BRIEF DESCRIPTION OF DRAWINGS

[0021] Figure 1 Cell migration was evaluated for wound healing assay in L929 fibroblast cells treated with compound A5 for 0h, 24h and 48h, images were observed under an inverted microscope with phase contrast, wherein (A) is the wound healing of UVB damaged HaCat cells after treatment with compound A5, (B) is the statistical result of the percentage of wound healing, the wound healing is expressed as: mean ± standard deviation (n = 3), (P < 0.05);

[0022] Figure 2 A5 for repair of UVB-induced skin damage in mice, wherein (A) is the record of mouse skin, (B) is the H&E staining of mouse skin (100x), (C) is the body weight of mice, (D) is the moisture analysis of mouse skin tissue content, NC (normal group, no ultraviolet irradiation), Model (model group, ultraviolet irradiation), Control (control group, empty matrix), A5-L (containing 0.5% A5 cream), A5-H (containing 2% A5 cream), mean ± standard deviation (n = 10), (P < 0.05). DETAILED DESCRIPTION

[0023] The above and other technical features and advantages of the present application will be more apparent from the following detailed description taken in conjunction with the accompanying drawings.

[0024] Example 1

[0025] Preparation of 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid cyclopropyl ester (as shown below):

[0026]

[0027] Trolox (500 mg, 2.0 mmol) and anhydrous tetrahydrofuran (5.0 mL) were weighed into a 10 mL reaction tube, DMAP (24 mg, 0.2 mmol) and cyclopropyl alcohol (125 mg, 2.1 mmol) were added, and finally dicyclohexyl carbodiimide (DCC, 3.0 mmol) was added. The reaction was carried out at room temperature until the Trolox reaction was complete. After dissolving in ethyl acetate (40 mL), it was washed with saturated aqueous sodium bicarbonate solution (25 mL) and saturated aqueous sodium chloride solution (25 mL) twice, respectively. After drying with anhydrous sodium sulfate and suction filtration, the solvent was evaporated under reduced pressure, and the target compound A1 was purified by silica gel column chromatography. White powder, yield 87%, melting point: 130.5-131.2°C.

[0028] 1H NMR (600 MHz, DMSO-d6) δ 7.48 (s, 1H, OH), 4.02 (m, 1H, OCH, cyclopropyl), 2.56 (m, 1H), 2.35 (m, 1H), 2.27 (m, 1H), 2.06 (s, 3H), 2.02 (s, 3H, CH3), 1.97 (s, 3H, CH3), 1.77 (m, 1H), 1.48 (s, 3H, CH3), 0.67-0.58 (m, 2H, cyclopropyl), 0.52-0.47 (m, 1H, cyclopropyl), 0.39-0.33 (m, 1H, cyclopropyl); TOF-HRMS: m / z [M+H] + calcd for C 17 H 23 O4: 291.1591; found: 291.1590.

[0029] Example 2

[0030] Preparation of 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid cyclopropyl methyl ester (as shown below):

[0031]

[0032] Trolox (500 mg, 2.0 mmol) and anhydrous tetrahydrofuran (5.0 mL) were weighed into a 10 mL reaction tube, DMAP (24 mg, 0.2 mmol) and cyclopropyl methyl alcohol (151 mg, 2.1 mmol) were added, and finally dicyclohexyl carbodiimide (DCC, 3.0 mmol) was added. The reaction was allowed to proceed at room temperature until the Trolox reaction was complete. After dissolving in ethyl acetate (40 mL), the solution was washed twice with saturated aqueous sodium bicarbonate solution (25 mL) and saturated aqueous sodium chloride solution (25 mL), respectively. After drying over anhydrous sodium sulfate, the solution was filtered under suction, and the solvent was evaporated under reduced pressure. The target compound A2 was obtained by silica gel column chromatography. White powder, yield 89%, melting point: 126.3-129.5 °C.

[0033] 1H NMR (500 MHz, CDC13) δ 7.35 (s, 1H, OH), 4.05 (m, 2H, OCH2), 2.76 (m, 1H), 2.58 (m, 1H), 2.43 (m, 1H), 2.26 - 2.15 (m, 7H), 2.09 (s, 3H), 1.63 - 1.58 (m, 1H, cyclopropyl), 1.51 (s, 3H, CH3), 0.64 (m, 2H, cyclopropyl), 0.56 (m, 2H, cyclopropyl); TOF-HRMS: m / z [M+H] + calcd for C 18 H 25 O4: 305.1748; found: 305.1753.

[0034] Example 3

[0035] Preparation of methyl ((R)-oxirane-2-yl) 6-hydroxy-2,5,7,8-tetramethylchroman-2- carboxylate (as shown below):

[0036]

[0037] Trolox (500 mg, 2.0 mmol) and anhydrous tetrahydrofuran (5.0 mL) were weighed into a 10 mL reaction tube, DMAP (24 mg, 0.2 mmol) and (R)-oxirane methanol (160 mg, 2.1 mmol) were added, and finally dicyclohexyl carbodiimide (DCC, 3.0 mmol) was added. The reaction was allowed to proceed at room temperature until the Trolox reaction was complete. After dissolving in ethyl acetate (40 mL), the solution was washed twice with saturated aqueous sodium bicarbonate solution (25 mL) and saturated aqueous sodium chloride solution (25 mL), respectively. After drying over anhydrous sodium sulfate, the solution was filtered, and the solvent was evaporated under reduced pressure. The target compound A3 was obtained by silica gel column chromatography. White powder, yield 83%, melting point: 128.6-130.5 °C.

[0038] 1 H NMR (400 MHz, CDC13) δ 5.45 (s, 1H), 4.37 (s, 1H), 4.34 (m, 1H), 3.62 - 3.56 (m, 1H), 3.30 (m, 1H), 3.08 (m, 1H), 2.97 (m, 1H), 2.94 (m, 1H), 2.43 (m, 1H), 2.19 - 2.16 (m, 4H), 2.11 (d, J = 8.1 Hz, 6H), 1.58 (s, 3H, CH3); TOF-HRMS: m / z [M+H] + calcd for C 17 H23 O5:307.1540; found: 307.1532.

[0039] Example 4

[0040] Preparation of methyl ((S)-oxirane-2-yl)-6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylate (as shown below):

[0041]

[0042] Trolox (500 mg, 2.0 mmol) and anhydrous tetrahydrofuran (5.0 mL) were weighed into a 10 mL reaction tube, DMAP (24 mg, 0.2 mmol) and (S)-oxirane methanol (160 mg, 2.1 mmol) were added, followed by dicyclohexyl carbodiimide (DCC, 3.0 mmol), and the reaction was allowed to proceed at room temperature until Trolox was completely reacted. The reaction mixture was dissolved in ethyl acetate (40 mL), washed with saturated aqueous sodium bicarbonate solution (25 mL) and saturated aqueous sodium chloride solution (25 mL) in turn, and dried over anhydrous sodium sulfate. After filtration, the solvent was evaporated under reduced pressure, and the target compound A4 was obtained by silica gel column chromatography. White powder, yield 91%, melting point: 127.2-129.2 °C.

[0043] 1 H NMR (500 MHz, CDC13) δ 5.29 (s, 1H), 4.61 (s, 1H), 4.45-3.84 (m, 2H), 2.59 (m, 4H), 2.17 (s, 2H), 2.10 (m, 9H, CH3), 1.61 (s, 3H, CH3); TOF-HRMS: m / z [M+H] + calcd for C 17 H 23 O5:307.1540; found: 307.1542.

[0044] Example 5

[0045] Preparation of cyclobutyl 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylate (as shown below):

[0046]

[0047] Trolox (500 mg, 2.0 mmol) and anhydrous tetrahydrofuran (5.0 mL) were weighed into a 10 mL reaction tube, DMAP (24 mg, 0.2 mmol) and cyclobutanol (155 mg, 2.1 mmol) were added, followed by dicyclohexylcarbodiimide (DCC, 3.0 mmol). The reaction was allowed to proceed at room temperature until Trolox was consumed. The reaction mixture was dissolved in ethyl acetate (40 mL) and washed twice with saturated aqueous sodium bicarbonate (25 mL) and saturated aqueous sodium chloride (25 mL), respectively. The organic phase was dried over anhydrous sodium sulfate and filtered. The solvent was removed under reduced pressure and the residue was purified by silica gel column chromatography to give the target compound A5. White powder, yield 82%, melting point: 144.7-146.2 °C.

[0048] 1 H NMR (500 MHz, CDC13) δ 4.97 - 4.88 (m, 1H, OCH), 4.47 (s, 1H, OH), 2.65 (ddd, J = 16.7, 6.3, 2.9 Hz, 1H), 2.53 (m, 1H), 2.43 (m, 1H), 2.35 - 2.25 (m, 2H), 2.20 (s, 3H, CH3), 2.14 (s, 3H, CH3), 2.05 (s, 3H, CH3), 2.00 (m, 1H, cyclobutyl-H), 1.92 - 1.84 (m, 2H, cyclobutyl-H), 1.79 - 1.72 (m, 1H, cyclobutyl-H), 1.61 (s, 4H, CH3); TOF-HRMS: m / z [M+H] + calcd for C 18 H 25 O4: 305.1747; found: 305.1749.

[0049] Example 6

[0050] Preparation of 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid-(3-oxacyclobutyl) ester (as shown below):

[0051]

[0052] Trolox (500 mg, 2.0 mmol) and anhydrous tetrahydrofuran (5.0 mL) were weighed into a 10 mL reaction tube, DMAP (24 mg, 0.2 mmol) and 3-oxetanemethanol (160 mg, 2.1 mmol) were added, followed by dicyclohexyl carbodiimide (DCC, 3.0 mmol), and the reaction was allowed to proceed at room temperature until Trolox was consumed. The reaction mixture was dissolved in ethyl acetate (40 mL), washed twice with saturated aqueous sodium bicarbonate (25 mL) and saturated aqueous sodium chloride (25 mL), dried over anhydrous sodium sulfate, and filtered. The solvent was removed under reduced pressure, and the residue was purified by silica gel column chromatography to give the target compound A6. White powder, yield 88%, melting point: 155.2-157.4 °C.

[0053] 1 H NMR (400 MHz, CDC13) δ 5.38 (p, J = 5.8 Hz, 1H, OCH), 4.88 - 4.79 (m, 2H, OCH2, oxetane), 4.55 (dd, J = 7.5, 5.4 Hz, 1H, OCH2, oxetane), 4.45 (dd, J = 7.4, 5.4 Hz, 1H, OCH2, oxetane), 4.33 (s, 1H, OH), 2.71 - 2.64 (m, 1H), 2.54 (dt, J = 12.4, 6.4 Hz, 1H), 2.50 - 2.41 (m, 1H), 2.19 (s, 3H, CH3), 2.16 (s, 2H), 2.06 (s, 3H, CH3), 1.91 (m, 1H), 1.64 (s, 4H, CH3); TOF-HRMS: m / z [M+H] + calcd for C 17 H 23 O5:307.1540; found:307.1539.

[0054] Example 7

[0055] Preparation of 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid cyclopentyl ester (as shown below):

[0056]

[0057] Trolox (500 mg, 2.0 mmol) and anhydrous tetrahydrofuran (5.0 mL) were weighed into a 10 mL reaction tube, DMAP (24 mg, 0.2 mmol) and cyclopentanol (181 mg, 2.1 mmol) were added, and finally dicyclohexylcarbodiimide (DCC, 3.0 mmol) was added. The reaction was allowed to proceed at room temperature until the Trolox reaction was complete. After dissolution in ethyl acetate (40 mL), the solution was washed twice with saturated aqueous sodium bicarbonate (25 mL) and saturated aqueous sodium chloride (25 mL), respectively. After drying over anhydrous sodium sulfate, the solution was filtered and the solvent was evaporated under reduced pressure. The target compound A7 was obtained by silica gel column chromatography. White powder, yield 90%, melting point: 135.6-137.3 °C.

[0058] 1 H NMR (400 MHz, CDC13) δ 7.06 (s, 1H), 4.87 (m, 1H, OCH), 2.83 (m, 1H), 2.65 (m, 1H), 2.45 (dd, J = 7.9, 5.1 Hz, 1H), 2.20 - 2.11 (m, 10H), 1.93 - 1.81 (m, 2H, cyclopentyl), 1.76 - 1.64 (m, 6H, cyclopentyl), 1.53 (s, 3H); TOF-HRMS: m / z [M+H] + calcd for C 19 H 27 O4: 319.1904; found: 319.1911.

[0059] Example 8

[0060] Preparation of 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid cyclohexyl ester (as shown below):

[0061]

[0062] Trolox (500 mg, 2.0 mmol) and anhydrous tetrahydrofuran (5.0 mL) were weighed into a 10 mL reaction tube, DMAP (24 mg, 0.2 mmol) and cyclopentanol (181 mg, 2.1 mmol) were added, and finally dicyclohexylcarbodiimide (DCC, 3.0 mmol) was added. The reaction was allowed to proceed at room temperature until the Trolox reaction was complete. After dissolution in ethyl acetate (40 mL), the solution was washed twice with saturated aqueous sodium bicarbonate (25 mL) and saturated aqueous sodium chloride (25 mL), respectively. After drying over anhydrous sodium sulfate, the solution was filtered and the solvent was evaporated under reduced pressure. The target compound A7 was obtained by silica gel column chromatography. White powder, yield 90%, melting point: 135.6-137.3 °C.

[0063] 1 H NMR (500 MHz, CDC13) δ 4.76 (m, 1H, OCH), 4.34 (d, J = 2.6 Hz, 1H, OH), 2.64 (m, 1H), 2.53 (m, 1H), 2.44 (m, 1H), 2.19 (s, 3H, CH3), 2.15 (s, 3H, CH3), 2.05 (s, 3H, CH3), 1.87 (m, 1H, cyclohexyl), 1.77 - 1.70 (m, 2H), 1.61 (s, 3H, CH3), 1.45 (m, 4H, cyclohexyl), 1.36 - 1.23 (m, 4H); TOF-HRMS: m / z [M+H] + calcd for C 20 H 29 O4: 333.2061; found: 333.2069.

[0064] Example 9

[0065] Preparation of 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid-(4- tetrahydropyranyl) ester (as shown below):

[0066]

[0067] Trolox (500 mg, 2.0 mmol) and anhydrous tetrahydrofuran (5.0 mL) were weighed into a 10 mL reaction tube, DMAP (24 mg, 0.2 mmol) and p-tetrahydropyranyl alcohol (215 mg, 2.1 mmol) were added, and finally dicyclohexyl carbodiimide (DCC, 3.0 mmol) was added. The reaction was allowed to proceed at room temperature until the Trolox reaction was complete. After dissolving in ethyl acetate (40 mL), the solution was washed twice with saturated aqueous sodium bicarbonate solution (25 mL) and saturated aqueous sodium chloride solution (25 mL), respectively. After drying over anhydrous sodium sulfate, the solution was filtered and the solvent was evaporated under reduced pressure. The target compound A9 was obtained by silica gel column chromatography. White powder, yield 76%, melting point: 123.5-125.3 °C.

[0068] 1 H NMR (400 MHz, CDC13) δ 6.97 (s, 1H), 4.82 (m, 1H, OCH), 3.79 (m, 2H, OCH2, pyran), 3.63 (m, 2H, OCH2, pyran), 2.87 (m, 1H), 2.82 (m, 1H), 2.65 (m, 1H), 2.20 - 2.11 (m, 10H), 2.03 (m, 2H, pyran), 1.82 (m, 2H, pyran), 1.53 (s, 3H); TOF-HRMS: m / z [M+H]+ calcd for C 19 H 27 O5: 335.1853; found: 335.1860.

[0069] Example 10

[0070] Preparation of 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid-(2- tetrahydropyran) ester (as shown below):

[0071]

[0072] Trolox (500 mg, 2.0 mmol) and anhydrous tetrahydrofuran (5.0 mL) were weighed into a 10 mL reaction tube, DMAP (24 mg, 0.2 mmol) and ortho-tetrahydropyranol (215 mg, 2.1 mmol) were added, followed by dicyclohexyl carbodiimide (DCC, 3.0 mmol), and the reaction was allowed to proceed at room temperature until Trolox was completely reacted. The reaction mixture was dissolved in ethyl acetate (40 mL), washed with saturated aqueous sodium bicarbonate solution (25 mL) and saturated aqueous sodium chloride solution (25 mL) in this order, and then dried over anhydrous sodium sulfate. After filtration, the solvent was evaporated under reduced pressure, and the target compound A10 was obtained by silica gel column chromatography. White powder, yield 81%, melting point: 126.3-128.5 °C.

[0073] 1 H NMR (400 MHz, CDC13) δ 7.08 (s, 1H), 5.73 (t, J = 3.4 Hz, 1H, OCH), 3.76 (m, 1H, pyran), 3.53 (m, 1H, pyran), 2.84 (m, 1H), 2.73 (m, 1H), 2.41 (m, 1H), 2.16 - 2.08 (m, 10H), 1.99 (m, 1H, pyran), 1.85 (m, 1H, pyran), 1.81 - 1.59 (m, 4H, pyran), 1.46 (s, 3H); TOF-HRMS: m / z [M+H] + calcd for C 19 H 27 O5: 335.1853; found: 335.1857.

[0074] Example 11

[0075] Preparation of 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid-(4- cyclohexanone) ester (as shown below):

[0076]

[0077] Trolox (500 mg, 2.0 mmol) and anhydrous tetrahydrofuran (5.0 mL) were weighed into a 10 mL reaction tube, DMAP (24 mg, 0.2 mmol) and p-hydroxycyclohexanone (240 mg, 2.1 mmol) were added, and finally dicyclohexyl carbodiimide (DCC, 3.0 mmol) was added. The reaction was allowed to proceed at room temperature until the Trolox reaction was complete. After dissolution in ethyl acetate (40 mL), the solution was washed twice with saturated aqueous sodium bicarbonate (25 mL) and saturated aqueous sodium chloride (25 mL), respectively. After drying over anhydrous sodium sulfate, the solution was filtered and the solvent was evaporated under reduced pressure. The target compound A11 was obtained by silica gel column chromatography. White powder, yield 86%, melting point: 134.5-136.2 °C.

[0078] 1 H NMR (400 MHz, CDC13) δ 6.99 (s, 1H), 4.79 (m, 1H, OCH), 2.93 (m, 1H), 2.85 (m, 1H), 2.58-2.43 (m, 1H), 2.41-2.37 (m, 1H), 2.31-2.25 (m, 2H, cyclohexanone), 2.22-2.15 (m, 1H), 2.17-2.08 (m, 12H), 1.97-1.83 (m, 2H, cyclohexanone), 1.39 (s, 3H); TOF-HRMS: m / z [M+H] + calcd for C 20 H 27 O5: 347.1853; found: 347.1846.

[0079] Example 12

[0080] Preparation of 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid-(4-(N- methylpiperidino)) ester (as shown below):

[0081]

[0082] Trolox (500 mg, 2.0 mmol) and anhydrous tetrahydrofuran (5.0 mL) were weighed into a 10 mL reaction tube, DMAP (24 mg, 0.2 mmol) and N-methyl-4-piperidinol (242 mg, 2.1 mmol) were added, and finally dicyclohexyl carbodiimide (DCC, 3.0 mmol) was added. The reaction was carried out at room temperature until the Trolox reaction was complete. After dissolving in ethyl acetate (40 mL), it was washed with saturated aqueous sodium bicarbonate solution (25 mL) and saturated aqueous sodium chloride solution (25 mL) twice, respectively, dried over anhydrous sodium sulfate, and then filtered under reduced pressure. The solvent was evaporated under reduced pressure and purified by silica gel column chromatography to obtain the target compound A12. White powder, yield 85%, melting point: 142.6-144.3°C.

[0083] 1 H NMR (400 MHz, CDC13) δ 7.10 (s, 1H), 4.83 (m, 1H, OCH), 3.09-2.95 (m, 3H), 2.92 (m, 1H), 2.66 (m, 2H, N-methylpiperidin), 2.45 (m, 1H), 2.27 (s, 3H), 2.20-2.11 (m, 10H), 2.06 (m, 2H, N-methylpiperidin), 1.68 (m, 2H, N-methylpiperidin), 1.48 (s, 3H); TOF-HRMS: m / z [M+H] + calcd for C 20 H 30 NO4: 348.2170; found: 348.2166.

[0084] I. Anti-inflammatory activity evaluation

[0085] 1. Experimental principle

[0086] The experiment uses the Griess reagent method, which is to use nitrite, sulfonic acid and azo salt to react to generate azo dye and detect its absorbance. Nitrite first reacts with sulfonic acid to generate nitroso sulfate, and then reacts with nitrogen salt to generate positively charged azo dye. This azo dye can be colored by visible light or ultraviolet spectrum and its absorbance can be measured to indirectly measure the concentration of nitrite.

[0087] 2. Experimental steps

[0088] (1) RAW264.7 cells in the logarithmic growth phase were plated in a 48-well plate, 50,000 cells per well, 300 μL per well, and incubated in an incubator for 24 h;

[0089] (2) Prepare culture solution with concentration gradient (50, 25, 12.5, 6.25, 3.125) by taking the original culture solution and adding culture solution containing drugs, set up blank group and stimulation group;

[0090] (3) After 1 h of drug treatment, add 30 μL LPS (0.5 μg / mL) and continue to culture for 24 h;

[0091] (4) Take out Grerss Reagent I and II reagent in advance, place them in natural conditions and restore to room temperature;

[0092] (5) At the end of drug treatment time, take out the 48-well plate, transfer the supernatant of cells in the wells to a 96-well plate, 50 μL per well;

[0093] (6) After transferring the liquid into the wells, add 50 μL of Griess Reagent I and 50 μL of Griess Reagent II;

[0094] (7) Transfer to the enzyme marker in the dark, detect the absorbance value at 540 nm wavelength;

[0095] (8) Calculate the NO inhibition rate and IC 50 according to the absorbance value.

[0096] 3. Experimental results

[0097] The experimental results are shown in Table 1.

[0098] Table 1: NO inhibition rate statistics table of corresponding numbered compounds in each example (IC 50 , μM)

[0099]

[0100]

[0101] Note: Each IC 50 value is the average value (±SEM) of at least 3 independent experiments.

[0102] II. Evaluation of antioxidant activity

[0103] 1. Experimental principle

[0104] ABTS method is a method widely used for detecting the antioxidant capacity of a substance in vitro. The method uses water-soluble ABTS+free radical initiator as a color developing agent, and ABTS reacts with potassium persulfate to generate stable blue / green cation radical ABTS+, the maximum absorption wavelength of which is 734 nm. When the measured substance is added to ABTS+, if there is an antioxidant component in the substance, the substance will react with ABTS+to cause the reaction system to fade, and then the change in absorbance at the 734 nm absorption wavelength of ABTS+free radicals is detected to reflect the antioxidant capacity of the substance.

[0105] 2. Experimental steps

[0106] (1) ABST solution and oxidant were mixed in equal volume to prepare a mother liquor;

[0107] (2) The mother liquor was placed at room temperature in the dark for 12-16 h;

[0108] (3) The mother liquor was diluted 35-55 times with 80% ethanol (or 30-50 times with PBS);

[0109] (4) The compound was prepared;

[0110] (5) 190 μL of the diluted ABTS solution was added;

[0111] (6) A standard curve of vitamin E (0.15, 0.3, 0.6, 0.9, 1.2, 1.5) was prepared;

[0112] (7) After adding the compound, the concentration was 0.5 μM;

[0113] (8) Incubate for 2-6 min;

[0114] (9) The absorbance was detected by an enzyme marker, and the A734 was about 0.7 or the A405 was about 1.4 after blank control with the corresponding PBS or 80% ethanol.

[0115] 3. Experimental results

[0116] The results are shown in Table 2:

[0117] Table 2 Antioxidant capacity of the corresponding numbered compounds in each example

[0118] Compound Trolox Equivalent Antioxidant Capacity (TEAC) A1 1.09±0.08 A2 1.58±0.05 A3 0.80±0.13 A4 0.68±0.02 A5 1.88±0.07 A6 1.17±0.01 A7 0.55±0.01 A8 1.01±0.05 A9 0.39±0.03 A10 0.61±0.01 A11 0.69±0.03 A12 1.08±0.01

[0119] As can be seen from Table 2, the 6-hydroxybenzopyran derivative in the application has good antioxidant activity and good application prospect and potential in skin anti-aging.

[0120] III. Wound healing experiment of compound A5

[0121] The 6-hydroxybenzopyran derivative A5 prepared in the above examples was selected to perform a cell scratch experiment.

[0122] 1. Experimental principle

[0123] Cell scratch assay, also known as wound healing assay, is a widely used experimental technique in cell biology research, mainly for evaluating cell migration ability, repair ability and cell-cell interaction. This experimental technique has important value in studying cell migration and invasion, wound healing process, and the effect of drugs on cell migration. The principle of cell scratch assay is based on artificially creating a "scratch" or blank area on a cell monolayer, and then observing and measuring how cells fill in the blank area. When the cell monolayer is confluent, a scratch is made on the cell layer using a gun head or other tools, and the cells at the edge of the scratch will fill in the blank area through migration. By capturing images at different time points and measuring the change in scratch distance, the migration speed and repair ability of cells can be quantitatively analyzed.

[0124] 2. Experimental steps

[0125] (1) Marking lines: Draw five horizontal lines as marking lines on the bottom of the six-well plate using a straight ruler and a marker pen;

[0126] (2) Plating: According to the grouping, use HaCaT cells with a cell density of about 5×10 5 cells per well, shake to evenly distribute the cells;

[0127] (3) Scratch: After the cells have grown (about 24h later), use a white gun head to draw a vertical line perpendicular to the well plate and the marking lines, so that the scratch intersects with the marking lines, forming 7 regions, and select 3 regions as fixed detection points;

[0128] (4) Washing: Discard the old culture medium and gently rinse with PBS for two to three times until the scraped cells are washed clean;

[0129] (5) Add liquid: According to the grouping, add compound I culture medium (6.25, 12.5μM) to the drug administration group, and add the same volume of serum-free culture medium to the control group;

[0130] (6) Photographing and observation: After the scratch, washing and liquid addition are completed, take 4X photos under the microscope as the 0h control. Place in a 37℃, 5% CO2 incubator for culture. At 24h and 48h time points, take out the cells and observe the scratch width at the same position under the microscope and take photos;

[0131] (7) Data analysis: ImageJ software can be used to analyze the scratch area, and the formula can be used for calculation.

[0132] 24h / 48h scratch area(%) = (0h scratch area-24h / 48h scratch area) ÷ 0h scratch area x 100

[0133] 3. Experimental results

[0134] The effect of compound A5 on enhancing HaCaT cell proliferation and migration was evaluated by scratch method, and the results are shown in Figure 1 The results show that the Normal group is slowly narrowed in 48 hours. Through drug intervention, at the concentration of 25 μM and 50 μM, the scratch is narrowed, the cell state is stable, and the change ratios of scratch area 0h-24h and 24h-48h are higher than those of the normal group, indicating that compound A5 has good repair ability and plays a certain curative effect on wound healing.

[0135] Four, preparation of 6-hydroxybenzopyran derivative-containing cream (taking compound A5 as an example)

[0136] The formulation prescription is shown in Table 3:

[0137] Table 3 Formulation prescription

[0138] Stearic acid 200g Petrolatum 100g Glyceryl monostearate 100g Liquid paraffin 300g Compound A5 120g Tween-80 100g Glycerol 200g Distilled water 5000 mL

[0139] Preparation process:

[0140] The oil phase stearic acid, vaseline, glycerol monostearate and liquid paraffin are mixed and melted at 80°C. Then compound A5 is mixed with the water phase Tween-80, glycerol, distilled water, and finally the water phase is mixed with the oil phase at 30°C and stirred for emulsification, to obtain the cream.

[0141] Five, protective effect of compound A5 in a mouse model of acute skin damage induced by ultraviolet rays

[0142] 1. Materials

[0143] The experimental model is 7-8 week old female BALB / c mice. They are provided by the Experimental Animal Center of Anhui Medical University and are raised in a 25±2°C clean and ventilated animal room under 12h light / dark conditions, 22-25°C ambient temperature, and free access to food and water.

[0144] 2. Animal experiment

[0145] Female BALB / c mice were randomly divided into 5 groups (n=10): (1) normal group (without UV irradiation); (2) model group (UV irradiation); (3) control group (base cream + UV irradiation); (4) low dose group (0.5% A5 cream + UV irradiation); (5) high dose group (2% A5 cream + UV irradiation). Mice were anesthetized by intraperitoneal injection of sodium pentobarbital (55 mg / kg) and then the hair on their backs was removed. After 24 hours of depilation, the depilatory cream was applied to the depilated area on the back for three consecutive days. On the fourth day, the mice were anesthetized again and a circular area with a diameter of 1 cm in the depilated area was irradiated with UV light at a wavelength of 311 nm for 30 minutes, while the rest of the body was shielded with tin paper; the drug was administered one hour after irradiation. The above-mentioned modeling method was performed twice a day, and UV irradiation was performed three times, and drug treatment was continued for four days after the last irradiation. The skin of the mice was photographed daily to record the depilated area. Finally, the skin of the mice was taken down to evaluate the water content of the skin components. According to the Chinese standard GB / T5009.3-2010, the dorsal samples were dried in an oven at 105°C for 4 hours until a stable weight was reached, thereby quantifying the water content of the skin. The formula for determining the water content is as follows: Water content (%) = (m a -m b ) / m a ×100%

[0146] where m a and m b represent the weight of the fresh finished skin sample and the dried finished skin sample, respectively.

[0147] 3. Experimental results

[0148] In the in vivo experiment, a UVB-induced mouse skin damage model was used to evaluate the anti-aging, moisturizing and repairing effects of A5. As shown in Figure 2 A, after the first UV irradiation, the mice showed mild skin damage and erythema symptoms, and after the second irradiation, the mice showed severe sunburn, skin damage and peeling symptoms, which were alleviated after preliminary treatment with A5-containing ointment. After completing three UV irradiations, treatment with A5-containing ointment significantly repaired the above symptoms. In Figure 2 B, epidermal density is an objective indicator for evaluating premature skin aging and inflammation. After staining, the skin of the mice exposed to UVB was observed to have a significantly increased epidermal density compared to the normal group. In contrast, the increase in epidermal thickness in mice exposed to UVB was alleviated after treatment with A5-containing ointment, especially with high concentrations of A5 ointment, and the treatment effect was more obvious. According to Figure 2As shown in C, the body weight of mice in the normal group kept a certain level of growth, but after UVB irradiation, the body weight of mice in the model group and the control group showed a downward trend; and the body weight of mice treated with the ointment containing A5 was improved after UVB-induced skin damage, thereby also indicating the safety and effectiveness of A5. It has been found through previous studies that the retention of water can enhance the defense ability of the skin. Figure 2 D illustrates the change of the water content in the skin of mice, and the use of the cream containing A5 can significantly improve the water retention ability of the skin due to the decrease of water content caused by ultraviolet irradiation, and has a moisturizing effect.

[0149] In summary, the 6-hydroxybenzopyran derivative prepared by the present application is an effective component for relieving inflammation, tissue repair and protection and treatment of ultraviolet damage. The compound has good anti-inflammatory, antioxidant activity and repair capacity, and can improve the skin condition, whiten, sunscreen and anti-aging, and has a comparable or even superior effect to special drugs, and has good application prospect and potential.

[0150] The above description is only the preferred embodiment of the present application, which is only illustrative but not limiting. It is understood by those skilled in the art that many changes, modifications and even equivalents can be made within the spirit and scope of the present application as defined in the claims, but all will fall within the protection scope of the present application.

Claims

1. A water-soluble vitamin E derivative, characterized by, The water-soluble vitamin E derivative is any one of the following: 、 、 、 、 、 。 2. A method for preparing the water-soluble vitamin E derivative according to claim 1, characterized by, The alcohol compound is any one of the following: cyclopropyl alcohol, cyclopropyl methyl alcohol, cyclobutyl alcohol, 3-hetero oxygen cyclobutyl alcohol, cyclohexyl alcohol, and 1-methyl-4-piperidyl alcohol.

3. The method of claim 2, wherein the water-soluble vitamin E derivative is prepared by the reaction of the vitamin E derivative of the formula (I) with the compound of the formula (II) in the presence of a base.

4. Use of the water-soluble vitamin E derivative of claim 1 in the preparation of an anti-inflammatory and anti-oxidation product. ​

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