Water-soluble vitamin E derivative as well as preparation method and application thereof

Abstract

The invention relates to the technical field of compound preparation, in particular to a water-soluble vitamin E derivative as well as a preparation method and application thereof.The parent nucleus is shown in the following formula: # imgabs0 #, R is naphthenic base or heterocyclic radical, the compound has good anti-inflammatory and anti-oxidation functions, collagen generation is promoted by inhibiting expression of inflammatory factors, and the anti-inflammatory and anti-oxidation effects are achieved. The anti-inflammatory and anti-aging effects are realized through ways of promoting the self-repairing of the skin barrier and the like, and a good application prospect is realized.

Description

Technical Field

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

[0002] Natural products are often used as the structural starting point for the design of new drugs, and structural transformation and modification are carried out on this basis to give the compounds new activities and functions. The structural modification of the natural product α-tocopherol (vitamin E) shows that by replacing the long-chain alkyl group on the side chain with a carboxyl group, a water-soluble vitamin E called Trolox is obtained. Trolox has been reported to have a wide range of biological activities, including anti-cancer, neuroprotection, and antioxidant. Trolox is also often used as a positive control to measure the antioxidant capacity of other antioxidants. By reducing oxidative stress, Trolox can increase wound healing ability; it has a protective effect on lipopolysaccharide-induced oxidative stress and inflammation by scavenging reactive oxygen species (ROS) and is a potential antioxidant against cellular oxidative damage.

[0003] When the skin is exposed to low-dose UVB radiation, intracellular ROS are generated, and at the same time, 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 redox balance of the skin and reducing cellular oxidative toxicity. However, excessive UVB radiation can lead to excessive accumulation of ROS, destroying the balance between the oxidation and antioxidant systems and causing severe oxidative stress. In addition, ROS-mediated oxidative stress can cause oxidative damage to a variety of biological macromolecules, such as proteins, DNA and lipids. This damage causes lipid peroxidation of cell membranes, promotes the release of lactate deoxygenase (LDH), reduces the levels of SOD, CAT and GSH, and increases the levels of lipid peroxides (LPO) and malondialdehyde (MDA). The p38 MAPK signaling pathway can be specifically activated by phosphorylation under the stress stimulation of UVB radiation and oxidative damage. The p38 MAPK signaling pathway is usually involved in the process of light damage and oxidative damage. In general, in response to 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 combat photodamage, but this self-repair is limited for severe photodamage. A single excessive exposure to UVB radiation can cause a variety of skin changes, including erythema, sunburn, vascular hyperpermeability, edema, pain, and fever, which are all characteristics of acute photodamage reactions and are usually closely related to oxidative stress. It can be seen that antioxidants play an important role in the repair process of acute photodamage. General antioxidant methods include applying sunscreens and using various antioxidants. However, most high-SPF sunscreens contain metal oxide nanoparticles and form a variety of organic components that absorb ultraviolet rays. The use of these sunscreens is limited because some of these ingredients are toxic and will 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 may 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, the development of new natural antioxidants to resist photodamage has attracted widespread attention and is very necessary.

[0004] In view of the above-mentioned defects, the inventors of the present invention finally obtained the present invention after a long period of research and practice. Summary of the invention

[0005] The purpose of the present invention is to solve the problem of how to prepare a new natural antioxidant that resists light damage, and provides a water-soluble vitamin E derivative, a preparation method and an application thereof.

[0006] In order to achieve the above object, the present invention discloses a water-soluble vitamin E derivative, wherein the water-soluble vitamin E derivative is a 6-hydroxybenzopyran derivative, and the parent nucleus of the 6-hydroxybenzopyran derivative is shown in the following formula:

[0007]

[0008] Wherein, 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 present invention also discloses a preparation method of the water-soluble vitamin E derivative, which is as follows: an alcohol compound and water-soluble vitamin E undergo a substitution reaction under alkaline conditions to obtain a 6-hydroxybenzopyran derivative.

[0012] The reaction process is as follows:

[0013]

[0014] Wherein, R is a cycloalkyl group or a heterocyclic group.

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

[0016] The invention also discloses the application of the water-soluble vitamin E derivative in preparing anti-inflammatory and anti-oxidative products.

[0017] Compared with the prior art, the beneficial effects of the present invention are:

[0018] 1. Simple preparation process, high recovery rate and easy operation;

[0019] 2. The water-soluble vitamin E derivatives prepared by the present invention are alicyclic esters or heterocyclic lipids, which retain the antioxidant activity of Trolox and enhance its anti-inflammatory activity. The synthetic compound has balanced antioxidant and anti-inflammatory activities and better oral bioavailability;

[0020] 3. The water-soluble vitamin E derivatives prepared by the present invention are effective ingredients in relieving inflammation, tissue repair and protection and treatment of ultraviolet damage. The compounds have good anti-inflammatory, antioxidant activity and repair ability, and have effects comparable to or even exceeding those of special drugs in improving skin condition, whitening, sun protection, anti-aging, etc., and have good application prospects and potential. BRIEF DESCRIPTION OF THE DRAWINGS

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

[0022] Figure 2 The repair effect of A5 on UVB-induced mouse skin damage, wherein, (A) is the mouse skin record, (B) is the H&E staining of the mouse skin (100×), (C) is the mouse body weight, (D) is the moisture analysis of the mouse skin tissue content, NC (normal group, no UV irradiation), Model (model group, UV 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 invention are described in more detail below 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 and placed in a 10 mL reaction tube, DMAP (24 mg, 0.2 mmol) and cyclopropanol (125 mg, 2.1 mmol) were added, and finally dicyclohexylcarbodiimide (DCC, 3.0 mmol) was added, and the reaction was carried out at room temperature until Trolox was completely reacted, and ethyl acetate (40 mL) was added to dissolve it, and then it was washed twice with saturated sodium bicarbonate aqueous solution (25 mL) and saturated sodium chloride aqueous solution (25 mL), dried over anhydrous sodium sulfate, and filtered, the solvent was evaporated under reduced pressure, and then purified by silica gel column chromatography to obtain the target compound A1. White powder, yield 87%, melting point: 130.5-131.2°C.

[0028] 1 H NMR (600 MHz, DMSO-d 6)δ7.48(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,CH 3 ),1.97(s,3H,CH 3 ),1.77(m,1H),1.48(s,3H,CH 3 ),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] + Calculate for C 17 H 23 O 4 :291.1591; found:291.1590.

[0029] Example 2

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

[0031]

[0032] Trolox (500 mg, 2.0 mmol) and anhydrous tetrahydrofuran (5.0 mL) were weighed and placed in a 10 mL reaction tube, DMAP (24 mg, 0.2 mmol) and cyclopropylmethanol (151 mg, 2.1 mmol) were added, and finally dicyclohexylcarbodiimide (DCC, 3.0 mmol) was added, and the mixture was reacted at room temperature until Trolox was completely reacted, and ethyl acetate (40 mL) was added to dissolve the mixture, and the mixture was washed twice with saturated aqueous sodium bicarbonate solution (25 mL) and saturated aqueous sodium chloride solution (25 mL), dried over anhydrous sodium sulfate, and filtered, and the solvent was evaporated under reduced pressure and purified by silica gel column chromatography to obtain the target compound A2. White powder, yield 89%, melting point: 126.3-129.5°C.

[0033] 1 H NMR (500 MHz, CDCl 3 )δ7.35(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,CH 3),0.64(m,2H,cyclopropyl),0.56(m,2H,cyclopropyl); TOF-HRMS:m / z[M+H] + calcdfor C 18 H 25 O 4 :305.1748; found:305.1753.

[0034] Example 3

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

[0036]

[0037] Trolox (500 mg, 2.0 mmol) and anhydrous tetrahydrofuran (5.0 mL) were weighed and placed in a 10 mL reaction tube, DMAP (24 mg, 0.2 mmol) and (R)-ethylene oxide methanol (160 mg, 2.1 mmol) were added, and finally dicyclohexylcarbodiimide (DCC, 3.0 mmol) was added, and the reaction was carried out at room temperature until Trolox was completely reacted, and ethyl acetate (40 mL) was added to dissolve it, and then it was washed twice with saturated sodium bicarbonate aqueous solution (25 mL) and saturated sodium chloride aqueous solution (25 mL) respectively, dried over anhydrous sodium sulfate, and filtered, the solvent was evaporated under reduced pressure, and then purified by silica gel column chromatography to obtain the target compound A3. White powder, yield 83%, melting point: 128.6-130.5°C.

[0038] 1 H NMR (400 MHz, CDCl 3 )δ5.45(s,1H),4.37(s,1H),4.34(m,1H),3.62–3.56(m,1H),3.30(m,1H),3.08(m,1H),2.9 7(m,1H),2.94(m,1H),2.43(m,1H),2.19–2.16(m,4H),2.11(d,J=8.1Hz,6H),1.58(s,3H,CH 3 ); TOF-HRMS: m / z[M+H] + Calculate for C 17 H 23 O 5 :307.1540; found:307.1532.

[0039] Example 4

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

[0041]

[0042] Trolox (500 mg, 2.0 mmol) and anhydrous tetrahydrofuran (5.0 mL) were weighed and placed in a 10 mL reaction tube, DMAP (24 mg, 0.2 mmol) and (S)-ethylene oxide methanol (160 mg, 2.1 mmol) were added, and finally dicyclohexylcarbodiimide (DCC, 3.0 mmol) was added, and the reaction was carried out at room temperature until Trolox was completely reacted, and ethyl acetate (40 mL) was added to dissolve it, and then it was washed twice with saturated sodium bicarbonate aqueous solution (25 mL) and saturated sodium chloride aqueous solution (25 mL), dried over anhydrous sodium sulfate, and filtered, the solvent was evaporated under reduced pressure, and then purified by silica gel column chromatography to obtain the target compound A4. White powder, yield 91%, melting point: 127.2-129.2°C.

[0043] 1 H NMR (500 MHz, CDCl 3 )δ5.29(s,1H),4.61(s,1H),4.45–3.84(m,2H),2.59(m,4H),2.17(s,2H),2.10(m,9H,CH 3 ),1.61(s,3H,CH 3 ); TOF-HRMS: m / z[M+H] + Calculate for C 17 H 23 O 5 :307.1540; found:307.1542.

[0044] Example 5

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

[0046]

[0047] Trolox (500 mg, 2.0 mmol) and anhydrous tetrahydrofuran (5.0 mL) were weighed and placed in a 10 mL reaction tube, DMAP (24 mg, 0.2 mmol) and cyclobutanol (155 mg, 2.1 mmol) were added, and finally dicyclohexylcarbodiimide (DCC, 3.0 mmol) was added, and the reaction was carried out at room temperature until Trolox was completely reacted, and ethyl acetate (40 mL) was added to dissolve it, and then it was washed twice with saturated sodium bicarbonate aqueous solution (25 mL) and saturated sodium chloride aqueous solution (25 mL), dried over anhydrous sodium sulfate, and filtered, the solvent was evaporated under reduced pressure, and then purified by silica gel column chromatography to obtain the target compound A5. White powder, yield 82%, melting point: 144.7-146.2°C.

[0048] 1 H NMR (500 MHz, CDCl 3 )δ4.97–4.88(m,1H,OCH),4.47(s,1H,OH),2.65(ddd,J=16.7,6.3,2.9Hz,1H),2.53(m,1H),2.43(m,1H),2.35–2.25(m,2H),2.20(s,3H,CH 3 ),2.14(s,3H,CH 3 ),2.05(s,3H,CH 3 ),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,CH 3 ); TOF-HRMS: m / z[M+H] + Calculate for C 18 H 25 O 4 :305.1747; found:305.1749.

[0049] Example 6

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

[0051]

[0052] Trolox (500 mg, 2.0 mmol) and anhydrous tetrahydrofuran (5.0 mL) were weighed and placed in a 10 mL reaction tube, DMAP (24 mg, 0.2 mmol) and 3-oxetane (160 mg, 2.1 mmol) were added, and finally dicyclohexylcarbodiimide (DCC, 3.0 mmol) was added, and the reaction was carried out at room temperature until Trolox was completely reacted, and ethyl acetate (40 mL) was added to dissolve it, and then it was washed twice with saturated sodium bicarbonate aqueous solution (25 mL) and saturated sodium chloride aqueous solution (25 mL) respectively, dried over anhydrous sodium sulfate, and filtered, the solvent was evaporated under reduced pressure, and then purified by silica gel column chromatography to obtain the target compound A6. White powder, yield 88%, melting point: 155.2-157.4°C.

[0053] 1 H NMR (400 MHz, CDCl 3 )δ5.38(p,J=5.8Hz,1H,OCH),4.88–4.79(m,2H,OCH 2 ,oxetane),4.55(dd,J=7.5,5.4Hz,1H,OCH 2 ,oxetane),4.45(dd,J=7.4,5.4Hz,1H,OCH 2 ,oxetane),4.33(s,1H,OH),2.71–2.64(m,1H),2.54(dt,J=12.4,6.4Hz,1H),2.50–2.41(m,1H),2.19(s,3H,CH 3 ),2.16(s,2H),2.06(s,3H,CH 3 ),1.91(m,1H),1.64(s,4H,CH 3 ); TOF-HRMS: m / z[M+H] + Calculate for C 17 H 23 O 5 :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 and placed in 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, and the reaction was carried out at room temperature until Trolox was completely reacted, and ethyl acetate (40 mL) was added to dissolve it, and then it was washed twice with saturated sodium bicarbonate aqueous solution (25 mL) and saturated sodium chloride aqueous solution (25 mL), dried over anhydrous sodium sulfate, and filtered, the solvent was evaporated under reduced pressure, and then purified by silica gel column chromatography to obtain the target compound A7. White powder, yield 90%, melting point: 135.6-137.3°C.

[0058] 1 H NMR (400 MHz, CDCl 3 )δ7.06(s,1H),4.87(m,1H,OCH),2.83(m,1H),2.65(m,1H),2.45(dd,J=7.9,5.1Hz,1H),2.20–2.11(m,1 0H),1.93–1.81(m,2H,cyclopentyl),1.76–1.64(m,6H,cyclopentyl),1.53(s,3H); TOF-HRMS:m / z[M+H] + Calculate for C 19 H 27 O 4 :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 and placed in a 10 mL reaction tube, DMAP (24 mg, 0.2 mmol) and cyclohexanol (215 mg, 2.1 mmol) were added, and finally dicyclohexylcarbodiimide (DCC, 3.0 mmol) was added, and the reaction was carried out at room temperature until Trolox was completely reacted, and ethyl acetate (40 mL) was added to dissolve it, and then it was washed twice with saturated sodium bicarbonate aqueous solution (25 mL) and saturated sodium chloride aqueous solution (25 mL) respectively, dried over anhydrous sodium sulfate, and filtered, the solvent was evaporated under reduced pressure, and then purified by silica gel column chromatography to obtain the target compound A8. White powder, yield 83%, melting point: 168.5-170.1°C.

[0063] 1 H NMR (500 MHz, CDCl 3 )δ4.76(m,1H,OCH),4.34(d,J=2.6Hz,1H,OH),2.64(m,1H),2.53(m,1H),2.44(m,1H),2.19(s,3H,CH 3 ),2.15(s,3H,CH 3 ),2.05(s,3H,CH 3 ),1.87(m,1H,cyclohexyl),1.77–1.70(m,2H),1.61(s,3H,CH 3 ),1.45(m,4H,cyclohexyl),1.36–1.23(m,4H); TOF-HRMS:m / z[M+H] + Calculate for C 20 H 29 O 4 :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 and placed in a 10 mL reaction tube, DMAP (24 mg, 0.2 mmol) and p-tetrahydropyranol (215 mg, 2.1 mmol) were added, and finally dicyclohexylcarbodiimide (DCC, 3.0 mmol) was added, and the reaction was carried out at room temperature until Trolox was completely reacted, and ethyl acetate (40 mL) was added to dissolve it, and then it was washed twice with saturated sodium bicarbonate aqueous solution (25 mL) and saturated sodium chloride aqueous solution (25 mL), dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated under reduced pressure and purified by silica gel column chromatography to obtain the target compound A9. White powder, yield 76%, melting point: 123.5-125.3°C.

[0068] 1 H NMR (400 MHz, CDCl 3 )δ6.97(s,1H),4.82(m,1H,OCH),3.79(m,2H,OCH 2 ,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] + Calculate for C 19 H 27 O 5 :335.1853; found:335.1860.

[0069] Example 10

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

[0071]

[0072] Trolox (500 mg, 2.0 mmol) and anhydrous tetrahydrofuran (5.0 mL) were weighed and placed in a 10 mL reaction tube, DMAP (24 mg, 0.2 mmol) and o-tetrahydropyranol (215 mg, 2.1 mmol) were added, and finally dicyclohexylcarbodiimide (DCC, 3.0 mmol) was added, and the reaction was carried out at room temperature until Trolox was completely reacted, and ethyl acetate (40 mL) was added to dissolve it, and then it was washed twice with saturated sodium bicarbonate aqueous solution (25 mL) and saturated sodium chloride aqueous solution (25 mL), dried over anhydrous sodium sulfate, and filtered, the solvent was evaporated under reduced pressure, and then purified by silica gel column chromatography to obtain the target compound A10. White powder, yield 81%, melting point: 126.3-128.5°C.

[0073] 1 H NMR (400 MHz, CDCl 3 )δ7.08(s,1H),5.73(t,J=3.4Hz,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] + Calculate for C 19 H 27 O 5:335.1853; found:335.1857.

[0074] Embodiment 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 and placed in a 10 mL reaction tube, DMAP (24 mg, 0.2 mmol) and p-hydroxycyclohexanone (240 mg, 2.1 mmol) were added, and finally dicyclohexylcarbodiimide (DCC, 3.0 mmol) was added, and the reaction was carried out at room temperature until Trolox was completely reacted, and ethyl acetate (40 mL) was added to dissolve it, and then it was washed twice with saturated sodium bicarbonate aqueous solution (25 mL) and saturated sodium chloride aqueous solution (25 mL) respectively, dried over anhydrous sodium sulfate, and filtered, the solvent was evaporated under reduced pressure, and then purified by silica gel column chromatography to obtain the target compound A11. White powder, yield 86%, melting point: 134.5-136.2°C.

[0078] 1 H NMR (400 MHz, CDCl 3 )δ6.99(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,cycl ohexanone),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] + Calculate for C 20 H 27 O 5 :347.1853; found:347.1846.

[0079] Example 12

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

[0081]

[0082] Trolox (500 mg, 2.0 mmol) and anhydrous tetrahydrofuran (5.0 mL) were weighed and placed in 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 dicyclohexylcarbodiimide (DCC, 3.0 mmol) was added, and the reaction was carried out at room temperature until Trolox was completely reacted, and ethyl acetate (40 mL) was added to dissolve it, and then it was washed twice with saturated sodium bicarbonate aqueous solution (25 mL) and saturated sodium chloride aqueous solution (25 mL), dried over anhydrous sodium sulfate, and filtered, the solvent was evaporated under reduced pressure, and then 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, CDCl 3 )δ7.10(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] + Calculate for C 20 H 30 NO 4 :348.2170; found:348.2166.

[0084] 1. Evaluation of anti-inflammatory activity

[0085] 1. Experimental Principle

[0086] The experiment uses the Griess reagent method, which is based on the reaction of nitrite, sulfonic acid and azo salt to generate azo dye and detect its absorbance. Nitrite first reacts with sulfonic acid to generate nitrososulfate, 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 nitrite concentration.

[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 grown naturally in an incubator for 24 h;

[0089] (2) Prepare a culture medium with a concentration gradient (50, 25, 12.5, 6.25, 3.125). Pipette the original culture medium, add the culture medium containing the drug, and set up a blank group and a stimulation group;

[0090] (3) 1 h after drug treatment, 30 μL LPS (0.5 μg / mL) was added and culture was continued for 24 h;

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

[0092] (5) After the drug treatment time is over, take out the 48-well plate and transfer the cell supernatant in the well to the 96-well plate, with 50 μL transferred to each well;

[0093] (6) After the liquid is transferred into the well, add 50 μL of Griess Reagent I and then 50 μL of Griess Reagent II;

[0094] (7) Transfer to a microplate reader away from light and measure the absorbance at a wavelength of 540 nm;

[0095] (8) Calculate NO inhibition rate and IC based on absorbance 50 .

[0096] 3. Experimental results

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

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

[0099]

[0100]

[0101] Note: Each IC 50 Values ​​are means (± SEM) of at least 3 independent experiments.

[0102] 2. Evaluation of Antioxidant Activity

[0103] 1. Experimental Principle

[0104] The ABTS method is a widely used method for detecting the antioxidant capacity of substances in vitro. This method uses a water-soluble ABTS+ free radical initiator as a color developer. ABTS reacts with potassium persulfate to generate a stable blue / green cationic free radical ABTS+ with a maximum absorption wavelength of 734nm. When the substance to be tested is added to ABTS+, if there are antioxidant components in the substance, the substance will react with ABTS+ to fade the reaction system, and then the change in absorbance is detected at the 734nm absorption wavelength of the ABTS+ free radical to reflect the antioxidant capacity of the substance.

[0105] 2. Experimental steps

[0106] (1) Mix equal volumes of ABST solution and oxidant to prepare a mother solution;

[0107] (2) The mother solution was placed at room temperature away from light for 12-16 hours;

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

[0109] (4) preparing the compound;

[0110] (5) Add 190 μL of diluted ABTS solution;

[0111] (6) Prepare a vitamin E standard curve (0.15, 0.3, 0.6, 0.9, 1.2, 1.5);

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

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

[0114] (9) The absorbance was measured by an ELISA reader. After blank control with corresponding PBS or 80% ethanol, A734 was about 0.7 or A405 was about 1.4.

[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 derivatives of the present invention have good antioxidant activity and show good application prospects and potential in skin anti-aging.

[0120] 3. Wound healing experiment of compound A5

[0121] The 6-hydroxybenzopyran derivative A5 prepared in the above example was selected for the cell scratch test.

[0122] 1. Experimental Principle

[0123] The cell scratch assay, also known as the wound healing assay, is an experimental technique widely used in cell biology research, mainly used to evaluate the migration ability, repair ability and cell-to-cell interaction of cells. This experimental technique is of great value in studying cell migration and invasion, the wound healing process, and the effects of drugs on cell migration. The principle of the cell scratch assay is based on artificially creating a "scratch" or blank area on the cell monolayer, and then observing and measuring how the cells fill this blank area. When the cell monolayer is fused, a scratch is made on the cell layer using a gun tip or other tool, and the cells at the edge of the scratch will fill this blank area by migrating. By capturing images at different time points and measuring the changes in the scratch spacing, the migration speed and repair ability of the cells can be quantitatively analyzed.

[0124] 2. Experimental steps

[0125] (1) Marking: On the bottom of the six-well plate, use a marker to draw five horizontal lines along the ruler as marking lines;

[0126] (2) Plating: Plating according to the group, using HaCaT cells, with a cell density of 5×10 5 About 100 cells / well, shake to spread the cells evenly;

[0127] (3) Scratching: After the cells have grown fully (about 24 hours later), use the tip of a white gun to draw a vertical line perpendicular to the well plate and the marked line so that the scratch and the marked line intersect, forming 7 areas. Select 3 areas as fixed detection points.

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

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

[0130] (6) Photographing and observation: After scratching, cleaning, and adding liquid, take 4X photos with a microscope as a 0 h control. Place in 37°C, 5% CO 2 Culture in an incubator. Take out the cells at 24h and 48h, observe the width of the scratch at the same position under a microscope and take photos;

[0131] (7) Data analysis: ImageJ software can be used to analyze the scratch area and calculate it using the formula.

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

[0133] 3. Experimental results

[0134] The effect of compound A5 on enhancing the proliferation and migration of HaCaT cells was evaluated by the scratch assay. Figure 1 As shown. The results showed that the scratches in the Normal group slowly narrowed within 48 hours. Through drug intervention, at concentrations of 25μM and 50μM, the scratches narrowed, the cell state was stable, and the scratch area 0h-24h and 24h-48h change ratios were higher than those in the normal group, indicating that compound A5 has good repair ability and has a certain therapeutic effect on wound healing.

[0135] 4. Preparation of cream containing 6-hydroxybenzopyran derivative (taking compound A5 as an example)

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

[0137] Table 3 Preparation prescription

[0138] Stearic acid 200g Vaseline 100g Glyceryl Monostearate 100g Liquid paraffin 300g Compound A5 120g Twain-80 100g glycerin 200g Distilled water 5000mL

[0139] Preparation process:

[0140] The oil phase of stearic acid, vaseline, glyceryl monostearate and liquid paraffin were mixed and melted at 80° C. Then, compound A5 was mixed with the aqueous phase of Tween-80, glycerin and distilled water, and finally the aqueous phase and the oil phase were mixed and stirred at 30° C. for emulsification to obtain a cream.

[0141] 5. Protective effect of compound A5 in a mouse model of UV-induced acute skin damage

[0142] 1. Materials

[0143] The experimental model was 7-8 week old female BALB / c mice, provided by the Experimental Animal Center of Anhui Medical University, and kept in a purified and ventilated animal room at 25±2℃, 12h light / dark conditions, 22-25℃ ambient temperature, with free access to food and water.

[0144] 2. Animal experiments

[0145] Female BALB / c mice were randomly divided into 5 groups (n=10): (1) normal group (no ultraviolet irradiation); (2) model group (ultraviolet irradiation); (3) control group (matrix cream + ultraviolet irradiation); (4) low-dose group (0.5% A5 cream + ultraviolet irradiation); (5) high-dose group (2% A5 cream + ultraviolet irradiation). The mice were anesthetized by intraperitoneal injection of sodium pentobarbital (55 mg / kg), and then the hair on the back was removed. After 24 hours of depilation, 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 ultraviolet light at an intensity of 311 nm for 30 minutes, while the rest of the body was covered with tin foil; the drug was administered one hour after irradiation. The above modeling method was repeated every two days, for a total of three ultraviolet irradiations, and the drug treatment was continued for four days after the last irradiation. The skin of the mouse depilated area was photographed and recorded daily. Finally, the skin of the mouse was removed to evaluate the water content of the skin components. According to China's GB / T5009.3-2010 standard, the back sample was dried in an oven at 105°C for 4 hours until a stable weight was reached to quantify the moisture content of the skin. The formula for determining the moisture content is as follows: Moisture content (%) = (m a -m b ) / m a ×100%

[0146] Among them, m a and m b Refers to the weight of fresh finished leather samples and dried finished leather samples respectively.

[0147] 3. Experimental results

[0148] In vivo experiments were conducted using a UVB-induced mouse skin damage model to evaluate the anti-aging, moisturizing, and repair effects of A5. Figure 2 As shown in A, after one exposure to ultraviolet light, the mice developed mild skin damage and erythema symptoms, while after the second exposure, the mice developed severe sunburn, skin damage and peeling symptoms. After initial treatment with an ointment containing A5, the above symptoms were alleviated. After completing three exposures to ultraviolet light, treatment with an ointment containing A5 significantly repaired the above symptoms. Figure 2 In B, epidermal density is an objective indicator for evaluating premature skin aging and inflammation. After observing the stained skin, the epidermal density of mice irradiated with UVB increased significantly compared with the normal group. In contrast, the increase in epidermal thickness of mice exposed to UVB was alleviated after using the ointment containing A5, especially when using a high concentration of A5 ointment, the therapeutic effect was more obvious. Figure 2As shown in Figure C, the weight of mice in the normal group has been increasing at a certain level, but after UVB irradiation, the weight of mice in the model group and the control group showed a downward trend; while the weight loss of mice treated with the ointment containing A5 was improved after UVB-induced skin damage, which also shows the safety and effectiveness of A5. According to previous studies, moisture retention can enhance skin defense capabilities. Figure 2 D shows the change in water content in mouse skin. Treatment with a cream containing A5 can significantly improve the skin's ability to lock in moisture that has been reduced due to ultraviolet radiation, and has a moisturizing effect.

[0149] In summary, the 6-hydroxybenzopyran derivatives prepared by the present invention are effective ingredients in relieving inflammation, tissue repair and protection and treatment of ultraviolet damage. The compound has good anti-inflammatory, antioxidant activity, and repair ability, and has effects comparable to or even exceeding those of special drugs in improving skin conditions, whitening, sun protection, and anti-aging, and has good application prospects and potential.

[0150] The above description is only a preferred embodiment of the present invention, which is only illustrative and not restrictive of the present invention. Those skilled in the art understand that many changes, modifications, and even equivalences may be made to the present invention within the spirit and scope defined by the claims of the present invention, but all of them will fall within the scope of protection of the present invention.

Claims

1. A water-soluble vitamin E derivative, characterized in that: The water-soluble vitamin E derivative is a 6-hydroxybenzopyran derivative, and the parent nucleus of the 6-hydroxybenzopyran derivative is shown in the following formula: Wherein, R is a cycloalkyl group or a heterocyclic group.

2. A water-soluble vitamin E derivative according to claim 1, characterized in that: The water-soluble vitamin E derivative is any one of the following:

3. A method for preparing a water-soluble vitamin E derivative as claimed in claim 1 or 2, characterized in that: The details are as follows: an alcohol compound and water-soluble vitamin E undergo a substitution reaction under alkaline conditions to obtain a 6-hydroxybenzopyran derivative.

4. The method for preparing a water-soluble vitamin E derivative according to claim 3, characterized in that: The reaction process is as follows: Wherein, R is a cycloalkyl group or a heterocyclic group.

5. The method for preparing a water-soluble vitamin E derivative according to claim 3, characterized in that: The alcohol compound is any one of cyclopropanol, cyclopropylmethanol, (S)-glycidol, (R)-glycidol, cyclobutanol, 3-heterocyclobutanol, cyclopentanol, cyclohexanol, 4-hydroxyfuran, 2-hydroxyfuran, 4-hydroxycyclohexanone and 1-methyl-4-piperidinol.

6. Use of the water-soluble vitamin E derivative according to claim 1 or 2 in preparing anti-inflammatory and antioxidant products.

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