Application of hederacoside C and deglycosylated derivative thereof in preparation of drugs or skin care products for UV damage repair

Abstract

The invention discloses application of hederacoside C and a deglycosylated derivative thereof in preparation of a medicine or a skin care product for UV (ultraviolet) injury repair, and cell experiments find that the hederacoside C and the deglycosylated derivative thereof have no obvious cytotoxicity to human immortalized keratinocytes and have a remarkable proliferation promoting effect; and the ROS level in cells after UV irradiation can be obviously reduced. Besides, animal experiment results show that the hederacoside C and the deglycosylated derivative thereof can effectively relieve symptoms such as red and swollen skin, rough skin, deepened wrinkles and reduced elasticity caused by UVB radiation, reduce the thickness increase of epidermis and spinous layers of mice, increase the content of collagenous fibers and enable the collagenous fibers to be arranged neatly, reduce the content of a proinflammatory factor IL-6 in tissues and improve the curative effect of the hederacoside C and the deglycosylated derivative thereof. The effect of repairing skin oxidation caused by UVB radiation is better than that of commercially available sunburn repair milk, and a scientific basis is provided for application of hederacoside C and deglycosylated derivatives thereof in preparation of drugs or skin care products for UV injury repair.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to the use of hederacoside C and its deglycosylated derivatives in the preparation of medicines or skin care products for repairing UV damage. Background Art

[0002] In recent years, with the increasing prevalence of skin damage caused by ultraviolet (UV) radiation, the search for safe and effective skin-repairing ingredients has become a research hotspot in the cosmetics and pharmaceutical fields. UV radiation, a component of sunlight, is divided into three types: UVA, UVB, and UVC, depending on its wavelength. Although UVB (280-320 nm) accounts for only approximately 5% of the UV rays reaching the Earth's surface, its high energy allows it to penetrate the epidermis and penetrate deep into the dermis, approximately 160-180 μm deep. This can directly and indirectly cause a range of skin problems, including redness, dryness, increased wrinkling, hyperpigmentation, decreased elasticity, skin aging, and even skin cancer. UVB radiation not only directly damages DNA in the skin but also induces oxidative stress, inducing the production of reactive oxygen species (ROS), disrupting cellular redox homeostasis. Furthermore, UVB radiation activates pro-inflammatory signaling pathways such as NF-κB, promoting the release of inflammatory factors and triggering skin inflammatory responses. Therefore, the development of active ingredients with antioxidant and anti-inflammatory properties, as well as those that promote skin barrier repair, holds great potential for application.

[0003] Currently, drugs and skincare products used to repair UVB damage primarily rely on the following ingredients: vitamin C, vitamin E, antioxidants, anti-inflammatory ingredients like dipotassium glycyrrhizinate, and DNA repair enzymes. These ingredients suffer from limitations such as limited efficacy, insufficient stability, and insufficient synergistic regulation of skin barrier repair and collagen regeneration. Natural products, due to their multi-target effects and favorable safety profile, have become an important source for the development of UVB repair drugs. Hederacoside C (HDC), an active ingredient derived from the leaves of the traditional medicinal plant Hedera helix L., has shown potential in the treatment of inflammatory diseases due to its significant anti-inflammatory, expectorant, and anti-tumor activities. However, no research has been conducted on its application in repairing UVB-induced skin damage. Summary of the Invention

[0004] To solve the above problems, the present invention provides the use of hederacoside C and its deglycosylated derivatives in the preparation of medicines or skin care products for UV damage repair. It is verified at the cellular level that hederacoside C and its deglycosylated derivatives have no obvious cytotoxicity to human immortalized keratinocytes, have a significant proliferation-promoting effect, and can significantly reduce the level of ROS in cells; at the same time, it is verified at the animal level that hederacoside C and its deglycosylated derivatives can effectively alleviate skin redness, roughness, deepening of wrinkles, decreased elasticity and other symptoms caused by UVB radiation, reduce the increase in the thickness of the mouse epidermis and spinous layer, increase the content of collagen fibers and arrange them neatly, and at the same time reduce the content of the pro-inflammatory factor IL-6 in the tissue.

[0005] Specifically, the following technical solutions are provided:

[0006] The first aspect of the present invention provides the use of hederacoside C and its deglycosylated derivatives in the preparation of drugs or skin care products for UV damage repair, wherein the deglycosylated derivatives of hederacoside C are compounds represented by formula P3 and / or formula P5:

[0007]

[0008] Furthermore, the structure of the hederacoside C (the numbers in the structure represent the carbon numbers at different positions) is shown below:

[0009]

[0010] Furthermore, the UV damage is skin damage caused by UVB radiation.

[0011] Furthermore, the deglycosylated derivative of hederacoside C is a compound represented by formula P5.

[0012] Furthermore, the medicine or skin care product is used to promote the proliferation of human immortalized keratinocytes.

[0013] Furthermore, the medicine or skin care product is used to reduce the level of ROS in cells after UV irradiation.

[0014] Furthermore, the medicine or skin care product is used to inhibit skin epidermal thickening and / or inflammatory infiltration caused by UV radiation.

[0015] Furthermore, the medicine or skin care product is used to promote the increase of collagen fiber content in the dermis and improve its arrangement structure.

[0016] Furthermore, the medicine or skin care product is used to reduce the expression level of the pro-inflammatory factor IL-6 in skin tissue.

[0017] Furthermore, the drug also includes a pharmaceutically acceptable carrier, which includes one or more of a diluent, a solubilizer, a cosolvent, a disintegrant, a dispersant, a lubricant, a flavoring agent, an antioxidant, a binder, an absorbent, a wetting agent, a buffer, and a cross-linking agent, such as cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), cyclodextrins (such as hydroxypropyl cyclodextrin), emulsifiers (such as Tween), wetting agents (such as sodium lauryl sulfate), colorants, flavorings, stabilizers, antioxidants, preservatives, pyrogen-free water, etc., including but not limited to the types of carriers listed above.

[0018] Furthermore, the skin care product also includes excipients acceptable in the field of skin care products, and the excipients include one or more of whitening agents, moisturizers, antioxidants, surfactants, preservatives, fragrances, and solvents; the solvents include water, alcohols, and the like.

[0019] Furthermore, the dosage form of the drug is ointment, patch, pill, tablet, powder, capsule, granule, liquid, injection, gel or suppository; the administration method of the drug includes oral administration, rectal administration, topical administration, transdermal administration, etc.; more preferably, transdermal administration.

[0020] Furthermore, the skin care product is a solution, suspension, emulsion, cream, paste, gel, soap, oil, dry powder, liquid foundation, wet powder or spray.

[0021] Furthermore, the drug can be administered alone or in combination with other therapeutic drugs; the skin care product can be administered alone or in combination with other skin care products.

[0022] Furthermore, the preparation method of the deglycosylated derivative of hederacoside C shown in formula P3 is as follows: hederacoside C is hydrolyzed with α-L-rhamnosidase A (RhaA) in the presence of water to obtain the deglycosylated derivative of hederacoside C shown in formula P3; the temperature of the hydrolysis reaction is 50-60°C and the time is 6-8h.

[0023] Furthermore, the preparation method of the deglycosylated derivative of hederacoside C shown in Formula P5 is as follows: hederacoside C is subjected to a primary hydrolysis reaction with α-L-rhamnosidase A (RhaA) in the presence of water, and is inactivated after the reaction is complete, and then β-glucosidase (BG) is added for a secondary hydrolysis reaction to obtain the deglycosylated derivative of hederacoside C shown in Formula P5; the temperature of the primary hydrolysis reaction is 50-60°C, and the time is 6-8h; the temperature of the secondary reaction is 50-60°C, and the time is 6-8h.

[0024] A second aspect of the present invention provides a cream for repairing skin damage caused by UV radiation, the cream comprising an active ingredient selected from one or more of hederacoside C and its deglycosylated derivatives, wherein the mass proportion of the active ingredient in the cream is 5%-8%.

[0025] The deglycosylated derivative of hederacoside C is a compound represented by Formula P3 and / or Formula P5:

[0026]

[0027] In some preferred embodiments of the present invention, the cream comprises the following components in parts by mass: 200 parts of active ingredient, 375 parts of glycerin, 2.5 parts of ethylparaben, 75 parts of Tween 80, 1847.5 parts of water, 275 parts of stearic acid, 187.5 parts of glyceryl monostearate, and 250 parts of vaseline.

[0028] Beneficial effects of the present invention:

[0029] The present invention provides the use of hederacoside C and its deglycosylated derivatives in the preparation of medicines or skin care products for UV damage repair. It is verified at the cellular level that hederacoside C and its deglycosylated derivatives have no obvious cytotoxicity to human immortalized keratinocytes, have a significant pro-proliferation effect, and can significantly reduce the level of ROS in cells after UVB irradiation. At the same time, it is verified at the animal level that hederacoside C and its deglycosylated derivatives can effectively alleviate the symptoms of skin redness, roughness, deepening of wrinkles, and decreased elasticity caused by UVB radiation, reduce the increase in the thickness of the mouse epidermis and spinous layer, increase the content of collagen fibers and arrange them neatly, and reduce the content of the pro-inflammatory factor IL-6 in the tissue. In addition, compared with hederacoside C, hederacoside C deglycosylated derivatives P3 and P5 are more effective in reducing the reactive oxygen species in cells after UVB radiation and repairing skin damage caused by UVB radiation.

[0030] The present invention describes the therapeutic effects and mechanism of hederacoside C and its deglycosylated derivatives on skin damage caused by UVB radiation, thereby providing a scientific basis for the preparation of drugs or skin care products for repairing UV damage using hederacoside C and its deglycosylated derivatives. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 The figure shows the cytotoxicity of hederacoside C and its deglycosylated derivatives to HaCaT cells. Compared with the blank group, **P<0.01, ***P<0.001, ****P<0.0001.

[0032] Figure 2This is a diagram showing the effects of hederacoside C and its deglycosylated derivatives on the changes of ROS in HaCaT cells after UVB irradiation;

[0033] Figure 3 This is a fluorescence intensity analysis of the changes in ROS in HaCaT cells after UVB irradiation by hederacoside C and its deglycosylated derivatives, compared with the blank group: ## P<0.01; compared with the model group: **P<0.01;

[0034] Figure 4 This is a diagram showing the effects of hederacoside C and its deglycosylated derivatives on the morphological changes of mouse skin after UVB irradiation;

[0035] Figure 5 This is an analysis chart of the IL-6 content in mouse skin tissue after ultraviolet B irradiation by hederacoside C and its deglycosylated derivatives, compared with the blank group: #### P < 0.0001; compared with the model group: **P < 0.01, ***P < 0.001, ****P < 0.0001;

[0036] Figure 6 This is a graph showing the effects of hederacoside C and its deglycosylated derivatives on the thickness of the epidermis and stratum spinosum of mice after UVB irradiation;

[0037] Figure 7 This figure shows the effects of hederacoside C and its deglycosylated derivatives on the epidermal collagen content of mice after UVB irradiation. DETAILED DESCRIPTION

[0038] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as commonly understood by those skilled in the art to which this invention pertains. The terms used in this specification of the present invention are for the purpose of describing specific embodiments only and are not intended to limit the present invention. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items. The terms "including" or "comprising" as used herein may also be replaced by the enclosed form "being" or "consisting of."

[0039] The present invention will be further described below with reference to the accompanying drawings and specific embodiments so that those skilled in the art can better understand the present invention and implement it. However, the embodiments are not intended to limit the present invention.

[0040] In the following examples, reagents were all existing products and were of analytical grade or higher. The high-performance liquid chromatograph (Agilent 1260) was manufactured by Agilent Technologies, Inc., USA, and the preparative liquid phase (HT7100A) used for compound purification was manufactured by Suzhou Huitong Separation and Purification Co., Ltd. NMR was recorded using a Bruker 600 MHz nuclear magnetic resonance spectrometer, and chemical shifts are expressed in δ (ppm). Data were statistically analyzed using one-way analysis of variance (ANOVA) and expressed as mean ± standard deviation (SD). P < 0.05 was considered statistically significant.

[0041] Example 1

[0042] This example relates to the preparation of a deglycosylated derivative P2-P5 of hederacoside C, and the specific operations are as follows:

[0043]

[0044] Preparation of the deglycosylated derivative P2 of hederacoside C: Using HDC as the raw material, a 20 mg / mL hederacoside C solution was prepared with deionized water. A 26 mg / mL RhaE solution was prepared using a pH = 6 buffer (made from disodium hydrogen phosphate and citric acid), which was filtered with a vacuum pump to obtain a clear solution. The two solutions were then mixed at a ratio of 1:1 (v / v) and reacted at 60°C for 2 hours. After the reaction, the mixture was inactivated in a boiling water bath for 20 minutes, an equal volume of methanol was added, and the supernatant was collected by centrifugation at 4500 rpm for 15 minutes. The supernatant was concentrated to one-tenth of the original volume, filtered through a 0.45 μm organic microporous filter membrane, and purified by preparative liquid phase separation to obtain product P2. Product P2 was characterized as follows:

[0045] 1H NMR(600MHz,DMSO-d6)δ5.21(dd,J=14.0,6.9Hz,3H),5.18–5.14(m,2H),5.11(s,1H),5.06(d ,J=7.4Hz,2H),4.91(d,J=16.2Hz,2H),4.82(d,J=4.6Hz,1H),4.68–4.58(m,4H),4.48–4.43( m,2H),4.43–4.39(m,1H),4.32(d,J=6.1Hz,1H),4.20(d,J=7.8Hz,1H),3.92(d,J=11.1Hz,2H ),3.76–3.68(m,4H),3.68–3.56(m,9H),3.55–3.41(m,12H),3.30(dd,J=27.3,11.1Hz,1H),3. 25–3.15(m,1H),3.10(d,J=8.3Hz,1H),3.08–2.99(m,1H),2.93(d,J=8.4Hz,1H),2.74(d,J=9 .0Hz,1H),1.99–1.91(m,1H),1.82–1.77(m,2H),1.76–1.67(m,2H),1.66–1.54(m,4H),1.52– 1.42(m,2H),1.38(d,J=9.3Hz,1H),1.34–1.29(m,1H),1.16(t,J=11.8Hz,4H),1.08(dt,J=11 .6, 6.4Hz, 4H), 0.94 (d, J = 13.4Hz, 1H), 0.86 (dd, J = 9.9, 7.4Hz, 5H), 0.68 (s, 1H), 0.56 (s, 1H).

[0046] 13 C NMR (151 MHz, DMSO-d6) gave 53 carbon signals, including δ: 175.6 (carbonyl carbon at position 28), 143.8, 122.0 (C-13, C-12, double bond carbon, showing two olefinic carbons sp 2Hybridization signal), 104.9, 103.3, 100.2, 94.3 (there are four sugar end group signals), 79.6 (C-3) is the carbon signal connected to the sugar on the aglycone, in the high field area 38.6 (C-1), 25.6 (C-2), 42.7 (C-4), 46.3 (C-5), 18.1 (C-6), 33.1 (C-7), 40.3 (C-8), 47.4 (C-9), 36.3 (C-10), 41.6 The carbonyl signals of the following compounds are as follows: (C-14), 27.5 (C-15), 22.8 (C-16), 41.0 (C-18), 45.9 (C-19), 30.6 (C-20), 33.5 (C-21), 32.1 (C-22), 63.6 (C-23), 13.4 (C-24), 17.0 (C-25), 17.4 (C-26), 25.9 (C-27), 33.5 (C-29), 23.7 (C-30). Among them, 33.5, 25.9, 23.7, 18.1 (C-11'), 17.4, 17.0 and 13.4 are 7 methyl carbon signals.

[0047] Preparation of the deglycosylated derivative P3 of hederacoside C: Using HDC as the raw material, a 20 mg / mL hederacoside C solution was prepared with deionized water. A 20 mg / mL RhaA solution was prepared using a pH 6 buffer (made from disodium hydrogen phosphate and citric acid), and the rhamnosidase solution was filtered using a vacuum pump to obtain a clear solution. The two solutions were then mixed at a ratio of 1:1 (v / v) and reacted at 60°C for 8 hours. After the reaction, the mixture was inactivated in a boiling water bath for 20 minutes, an equal volume of methanol was added, and the supernatant was collected by centrifugation at 4500 rpm for 15 minutes. The supernatant was concentrated to one-tenth of the original volume, filtered through a 0.45 μm organic microporous filter membrane, and purified by preparative liquid phase separation to obtain product P3. Product P3 was characterized as follows:

[0048] 1H NMR (600MHz, DMSO-d6) δ5.21(t,J=8.3Hz,3H),5.18–5.14(m,2H),4.90(s,1H),4.18(dt,J=10.1,7.4Hz,3H),3.91(d,J=11.3Hz,3H),3.67–3.55( m,16H),3.48(d,J=3.7Hz,8H),3.47–3.38(m,38H),3.34(dd,J=18.5,9.3 Hz,1H),3.31–3.26(m,1H),3.25–3.14(m,1H),3.14–2.99(m,1H),2.96–2 .90(m,1H),2.87(d,J=6.7Hz,1H),2.70(d,J=14.4Hz,1H),1.95(dt,J=1 3.6,6.8Hz,1H),1.87(s,1H),1.83–1.76(m,1H),1.70(dt,J=10.6,8.1Hz ,1H),1.65–1.41(m,3H),1.35(d,J=46.0Hz,1H),1.24–1.11(m,3H),1.07 (s,3H),0.99–0.92(m,1H),0.88–0.79(m,3H),0.67(s,2H),0.57(s,1H).

[0049] 13 C NMR (151 MHz, DMSO-d6) gave 53 carbon signals, δ175.4 (C-28) at low field was the carbon signal of the carboxyl group in the structure, δ121.8 (C-12) and δ143.6 (C-13) showed two alkenyl signals, which were the carbon signals of the double bonds at positions 12 and 13 in the structure, 105.3, 103.4, 94.5 (three sugar end group carbon signals), 80.2 (C-3), 77.2, 77.2, 77.1, 76.9, 73.9, 73.2, 72.7, 73.7 1.5,70.4,69.6,68.2,68.1,65.5,63.1,61.6,61.4,47.6,46.5,46.4,45.9,42.8,41.7,41.1,40.4,39.5,39.3,38.5,36.4,33.7,33.2,32.3,32.1,30.7,27.7,26.0,25.6,23.9,23.4,22.9,17.6,17.2,16.1,13.4. This structure contains only three sugar terminal carbon signals, no rhamnosyl methyl proton signal, and no methyl carbon signal at δ18.1.

[0050] Preparation of the deglycosylated derivative P4 of hederacoside C: Using HDC as the raw material, a 20 mg / mL hederacoside C solution was prepared in deionized water. A 26 mg / mL RhaE solution was prepared using a pH 6 buffer (made from disodium hydrogen phosphate and citric acid). The rhamnosidase solution was vacuum filtered to obtain a clear solution. The two solutions were then mixed in a 1:1 (v / v) ratio, reacted at 60°C for 2 hours, and inactivated in a boiling water bath for 20 minutes. β-glucosidase was added to a β-glucosidase concentration of 80 mg / mL. The reaction was continued for 8 hours. After the reaction was completed, an equal volume of methanol was added to terminate the reaction. The supernatant was centrifuged at 4500 rpm for 15 minutes and concentrated to one-tenth of the original volume. After filtration through a 0.45 μm organic microporous filter, the supernatant was purified by preparative liquid phase separation to obtain product P4. Product P4 was characterized as follows:

[0051] 1 H NMR (600MHz, DMSO-d6) δ5.21(t,J=8.3Hz,3H),5.18–5.14(m,2H),4.90(s,1H),4.18(dt,J=10.1,7.4Hz,3H),3.91(d,J=11.3Hz,3H),3.67–3.55( m,16H),3.48(d,J=3.7Hz,8H),3.47–3.38(m,38H),3.34(dd,J=18.5,9.3 Hz,1H),3.31–3.26(m,1H),3.25–3.14(m,1H),3.14–2.99(m,1H),2.96–2 .90(m,1H),2.87(d,J=6.7Hz,1H),2.70(d,J=14.4Hz,1H),1.95(dt,J=1 3.6,6.8Hz,1H),1.87(s,1H),1.83–1.76(m,1H),1.70(dt,J=10.6,8.1Hz ,1H),1.65–1.41(m,3H),1.35(d,J=46.0Hz,1H),1.24–1.11(m,3H),1.07 (s,3H),0.99–0.92(m,1H),0.88–0.79(m,3H),0.67(s,2H),0.57(s,1H).

[0052] 13C NMR (151 MHz, DMSO-d6) δ175.5 (C-28), 143.8 (C-13), 122.0 (C-12), 103.3, 100.2, 94.4 (three sugar end carbon signals), 79.6, 78.1, 77.0, 74.5, 73.3, 72.7, 72.3, 70.8, 70.7, 69.8, 68.4, 68.2, 64.8, 6 2.7,61.0,47.4,46.5,46.2,45.8,42.6,41.6,41.1,40.3,39.5,39.2,38.6,36.3,33.5,33.1,32.1,31.9,30.6,27.5,25.9,25.6,23.7,23.3,22.8,18.1,17.4,17.0,15.9,13.4.

[0053] Preparation of the deglycosylated derivative P5 of hederacoside C: Using HDC as the raw material, a 20 mg / mL hederacoside C solution was prepared in deionized water. A 20 mg / mL RhaA solution was prepared using a pH 6 buffer (made from disodium hydrogen phosphate and citric acid). The rhamnosidase solution was vacuum filtered to obtain a clear solution. The two solutions were then mixed in a 1:1 (v / v) ratio, reacted at 60°C for 8 hours, and inactivated in a boiling water bath for 20 minutes. β-glucosidase was added to a β-glucosidase concentration of 80 mg / mL. The reaction was continued for 8 hours. After the reaction was completed, an equal volume of methanol was added to terminate the reaction. The supernatant was centrifuged at 4500 rpm for 15 minutes and concentrated to one-tenth of the original volume. After filtration through a 0.45 μm organic microporous filter, the supernatant was purified by preparative liquid phase separation to obtain product P5. Product P5 was characterized as follows:

[0054] 1H NMR(600MHz,DMSO-d6)δ5.23(d,J=8.1Hz,2H),5.18–5.14(m,1H),4.18(d,J=5.6Hz,1H),3.68–3.54(m,5H),3.51–3.35(m,5H),3.30(ddd,J=14.2,7.7,4.0Hz,3H),3.19(dd,J=16.8,8.0Hz,1H),3.16–3.05(m,1H),2.73(dd,J=13.8,4.7Hz,1H),1.99–1.91(m,2H),1.87(s,2H),1.79(dd,J=9.9,3.4Hz,2H),1.77–1.61(m,3H),1.61–1.52(m,3H),1.52–1.48(m,2H),1.47(d,J=2.8Hz,1H),1.37(dd,J=21.8,18.6Hz,1H),1.24–1.11(m,5H),1.07(d,J=6.3Hz,2H),0.97–0.91(m,2H),0.87(d,J=6.9Hz,3H),0.68(s,2H),0.57(s,1H).

[0055] 13C NMR (151 MHz, DMSO-d6) gave 41 carbon signals. At low field, δ175.4 (C-28) was the carbon signal of the carboxyl group in the structure. δ121.8 (C-12) and δ143.6 (C-13) showed two alkenyl signals, which were the carbon signals of the double bonds at positions 12 and 13 in the structure. δ105.0 and 94.2 were the carbon signals of the two sugar end groups. 79.9 (C-3) was the carbon signal of the sugar connected to the aglycone. In the high field region, δ: 38.2 (C-1), 25.7 (C-2), 42.5 (C-4), 46.1 (C-5), 32.9 (C-7), 39.8 (C-8), 47.3 (C-9), 36.1 (C-10), 41.5 (C-14), 27.4 (C-15), 22. 7(C-16), 46.2(C-17), 40.9(C-18), 45.7(C-19), 30.5(C-20), 13.1(C-24), 23 .7(C-30), 16.9(C-25), 17.3(C-26), 25.9(C-27), 175.4(C-28), 33.4(C-29), 2 3.5 (C-30), 105.0 (C-1'), 94.2 (C-31), 77.9 (C-33), 76.8 (C-32), 72.9 (C-3'), 72.5 (C-35), 71.2 (C-2'), 69.7 (C-34), 67.8 (C-4'), 65.2 (C-5'), 62.8 (C-36). This structure contains only two sugar terminal carbon signals and no methyl carbon signal at δ18.1.

[0056] Example 2

[0057] In this example, cytotoxicity tests were performed on HDC and the hederacoside C deglycosylated derivatives P2, P3, P4, and P5 prepared in Example 1. The specific procedures were as follows:

[0058] The density of HaCaT cells in the logarithmic growth phase was adjusted to 1×10 5 / mL, 100μL of each well was inoculated into a 96-well plate, and the periphery of the plate was blocked with PBS. After the cells were completely attached overnight, the culture medium was aspirated and replaced with culture medium containing 200, 100, 50, 20, 10, and 5μg / mL sample solutions and incubated for 24 hours. The cell viability was then detected using a CCK-8 kit. 100μL of ten-fold diluted CCK-8 was added to each well, incubated in an incubator for 1-2 hours, and the absorbance was measured at 450nm on a microplate reader. The cell viability was calculated as follows:

[0059]

[0060] Where A0 is the absorbance value of the blank group, A1 is the absorbance value of the normal group, and A2 is the absorbance value of the sample group.

[0061] The test results are as follows Figure 1 As shown, except for compound P2, the cell survival rates of the other compounds after administration to HaCaT cells were all greater than 90%. Among them, the cell survival rates of components P3 and P5 were significantly higher than 100% at high concentrations, indicating that they were non-cytotoxic and exhibited a good proliferation-promoting effect.

[0062] Example 3

[0063] Ultraviolet radiation irradiates the skin, generating a large amount of ROS. The accumulation of excessive ROS can induce oxidative stress, thereby disrupting the normal skin condition. This example studies the effects of HDC and the deglycosylated derivatives of hederacoside C, P3 and P5, prepared in Example 1, on the reactive oxygen species content in HaCaT cells after ultraviolet radiation. The specific procedures are as follows:

[0064] HaCaT cells in the logarithmic growth phase were cultured at a rate of 5×10 5 After the cells were completely attached to the wall, the culture medium was removed and replaced with 1 mL of PBS to cover the bottom. The cells were then placed under UVB light for 5 minutes (irradiation dose of 162 mJ × cm -2 ) After irradiation, the solution was replaced with sample solutions containing 200, 100, and 50 μM. After incubation for another 24 hours, 10 μM DCFH-DA working solution (prepared in serum-free medium) was added to each well. The cells were then incubated in the dark in an incubator for 20 minutes and rinsed three times with serum-free medium. After treatment, the cells were photographed using an inverted fluorescence microscope, with five images taken per group. Fluorescence intensity was measured using Image J software.

[0065] The test results are as follows Figure 2 and Figure 3 As shown in the figure, compared with the normal group, the fluorescence intensity of the model group cells after UVB ultraviolet light irradiation increased significantly, while HaCaT cells administered with HDC, P3 and P5 respectively could significantly reduce this effect. Moreover, compared with the HDC group, the fluorescence intensity reduction effect of the P3 and P5 groups was more significant, indicating that P3 and P5 can effectively inhibit the increase of intracellular ROS content induced by UVB radiation and maintain the cellular redox balance.

[0066] Example 4

[0067] This example involves the preparation of different cream samples and the study of the effects of different cream samples on the epidermal appearance, inflammatory factors, and tissue of UVB-irradiated mice, as follows:

[0068] Different cream samples were weighed according to the proportions in the following table:

[0069]

[0070] The aqueous phase samples were mixed in a beaker, heated with stirring at 65°C until completely melted, and kept warm. The oil phase samples were mixed in a beaker, stirred at 65°C until completely melted, and kept warm. Using a glass rod to drain, the aqueous phase was slowly added to the oil phase while continuing to stir. Heating was stopped, and the mixture was cooled to room temperature while stirring until the cream was formed. This resulted in a blank group and creams for each sample group.

[0071] Construction of UVB radiation mouse model and intervention method: SPF Balb / c male mice, weighing 22±4g and aged 4-5 weeks, were randomly divided into 7 groups after one week of adaptive feeding, with 6 mice in each group, namely normal group, model group, blank cream group (negative control group), positive control group (Jiayankang sunburn repair cream), HDC group, P3 group, and P5 group. Before the formal experiment, the back of the mice was shaved with a shaver and depilatory cream, and then UVB irradiation was performed. The irradiation method was as follows: the mice in the model group, blank cream group, positive control group, HDC group, P3 group, and P5 group were placed 40cm under a UVB ultraviolet lamp (peak value at 313nm), irradiated for 1 hour every day for 6 consecutive days, and the total irradiation dose (irradiation dose = UVB irradiation intensity × time) was 2.9J×cm -2 Within 30 minutes after each irradiation, 60 mg of each cream was applied to the back skin of mice in the blank, positive, HDC, P3, and P5 groups. Twenty-four hours after the final irradiation and drug application, the mice were anesthetized and sacrificed by cervical dislocation, and skin tissue from the depilated back area was collected.

[0072] (1) Observation of mouse skin appearance

[0073] During the modeling and drug administration period, photographs of the experimental animals in each group were taken on days 0, 2, 4, and 6 after irradiation, and the recovery of the back skin was recorded.

[0074] The experimental results are as follows Figure 4 As shown, UVB irradiation caused a series of skin reactions on the back skin of mice, manifested as redness, swelling, desquamation, wrinkles, roughness, and dull luster. After 6 consecutive days of administration, the NC and PC groups still showed obvious redness, swelling, deepened wrinkles, and roughness. The HDC, P3, and P5 groups showed significant symptom relief, with slight redness and swelling and reduced wrinkles. Among them, the P5 group showed the best effect in repairing UVB radiation damage. These results indicate that, as evaluated by the appearance of mouse skin, hederacoside C and its derivatives can repair UVB radiation damage to mouse skin to a certain extent.

[0075] (2) Analysis of pro-inflammatory factors in skin tissue

[0076] Long-term and repeated exposure to ultraviolet rays will increase the expression level of IL-6 in local skin tissue. Excessive release of IL-6 will aggravate local inflammatory reactions and cause redness and swelling of the skin. Therefore, the IL-6 content in skin tissue is tested to reflect the inflammation of the skin tissue. The specific operation is as follows:

[0077] The collected skin tissue from the depilated back area was washed with physiological saline, dried with filter paper, weighed, added with physiological saline, homogenized in an ice bath, and centrifuged at 10,000 rpm for 5 minutes. The supernatant of the mouse skin tissue homogenate was collected and the IL-6 cytokine content was determined according to the instructions of the IL-6 kit.

[0078] The experimental results are as follows Figure 5 As shown in the results, compared with the normal group, the IL-6 content in the skin tissue of the model group was significantly increased, and the IL-6 in the mice of the PC, HDC, P3 and P5 groups showed a significant downward trend; compared with the HDC group, the decrease in IL-6 in the P3 and P5 groups was more obvious, indicating that the deglycosylated derivative of ivy glycoside C can reduce the release of the pro-inflammatory factor IL-6 caused by UVB radiation.

[0079] It can be seen that in the later stage of wound healing, compared with HDC and positive control drugs, P3 and P5 can better inhibit the release of proinflammatory cytokines, show better effects in eliminating inflammation, and are more conducive to wound healing.

[0080] (3) Histological analysis of mice

[0081] HE staining allows the nuclei, cytoplasm, and extracellular matrix in tissues to be clearly displayed, allowing for observation of internal tissue morphology. Therefore, mouse skin tissue was fixed with a tissue fixative for 24 hours, then embedded in paraffin and sectioned. Hematoxylin-eosin (HE) staining was then used to observe skin damage under a microscope.

[0082] The results are as follows Figure 6 As shown in the figure, in the skin of mice in the normal group, the skin texture was orderly arranged, and the collagen fibers were dense and neatly distributed. Compared with the normal group, the epidermis of the mice in the model group was hyperkeratinized, the spinous layer was significantly thickened, the collagen fibers were broken, and there was even inflammatory cell infiltration, all of which indicated that the skin had been severely damaged. After applying the medicated cream, it was found that the skin condition of mice in the NC group, PC group, and HDC group was slightly improved compared with the model group, but the epidermis still showed significant thickening. In contrast, the thickness of the epidermis and spinous layer of mice in the P3 and P5 groups was significantly reduced, and the collagen fibers were more neatly arranged. This shows that P3 and P5 can effectively inhibit the epidermal thickening and inflammatory infiltration of mouse skin caused by UVB radiation.

[0083] Collagen fibers, a key component of the dermis, are not only a hallmark of skin aging but also a key manifestation of photodamage. Therefore, Masson staining was used to assess changes in collagen fiber structure and content in damaged mouse skin. The procedure was as follows: Skin tissue was fixed with a tissue fixative for 24 hours, then embedded in paraffin and sectioned. The sections were then stained with Masson's trichrome. Skin damage was observed microscopically.

[0084] The results are as follows Figure 7 As shown in the figure, compared with the normal group, the blue-dyed collagen fiber structures in the skin of mice in the model, NC, and PC groups became lighter in color, with a reduced blue area and a disordered and irregular distribution in the skin, indicating that the collagen content in the skin of these mice was reduced and damaged. Compared with the model group, the collagen fiber content in the HDC group increased slightly, but the collagen fibers were locally loosely arranged. The collagen fiber content in the P3 and P5 groups increased significantly, with regular and dense arrangement. This suggests that P3 and P5 can alleviate the disordered collagen fiber structure and reduced collagen content in mice induced by UVB radiation.

[0085] In summary, it can be seen that hederacoside C and its deglycosylated derivatives P3 and P5 have no cytotoxicity and show a good proliferative effect on HaCaT cells. They can also effectively inhibit the increase in intracellular ROS content induced by UVB radiation and maintain cellular redox balance. And from the results of animal experiments, it can be seen that when hederacoside C or its deglycosylated derivatives P3 and P5 are topically applied in the form of a cream, hederacoside C or its deglycosylated derivatives P3 and P5 can significantly improve the apparent symptoms of skin redness, roughness, deepening of wrinkles and decreased elasticity caused by UVB radiation, effectively inhibit abnormal thickening of the epidermis and spinous layer, promote the increase in collagen fiber content in the dermis and improve its arrangement structure, and at the same time reduce the expression level of the pro-inflammatory factor IL-6 in skin tissue. It can be seen that hederacoside C and its deglycosylated derivatives P3 and P5 have good application prospects in the preparation of drugs or cosmetics for repairing skin damage caused by UVB radiation.

[0086] The above embodiments are merely preferred embodiments for the purpose of fully illustrating the present invention, and the scope of protection of the present invention is not limited thereto. Equivalent substitutions or modifications made by those skilled in the art based on the present invention are within the scope of protection of the present invention. The scope of protection of the present invention shall be subject to the claims.

Claims

1. Use of hederacoside C and its deglycosylated derivatives in the preparation of drugs or skin care products for UV damage repair, characterized in that: The deglycosylated derivative of hederacoside C is a compound represented by Formula P3 and / or Formula P5:

2. The use according to claim 1, characterized in that The UV damage is skin damage caused by UVB radiation.

3. The use according to claim 1, characterized in that The deglycosylated derivative of hederacoside C is a compound represented by formula P5.

4. The use according to claim 1, characterized in that The medicine or skin care product is used to promote the proliferation of human immortalized keratinocytes; The medicine or skin care product is used for reducing the intracellular ROS level after UV irradiation.

5. The use according to claim 1, characterized in that The medicine or skin care product is used to inhibit skin epidermal thickening and / or inflammatory infiltration caused by UV radiation; The medicine or skin care product is used for promoting the increase of the content of collagen fibers in the dermis and improving the arrangement structure thereof.

6. The use according to claim 1, characterized in that The medicine or skin care product is used to reduce the expression level of the pro-inflammatory factor IL-6 in skin tissue.

7. The use according to claim 1, characterized in that The drug further comprises a pharmaceutically acceptable carrier, which comprises one or more of a diluent, a solubilizer, a latent solvent, a disintegrant, a dispersant, a lubricant, a flavoring agent, an antioxidant, a binder, an absorbent, a wetting agent, a buffer, and a cross-linking agent.

8. The use according to claim 1, characterized in that The skin care product further comprises excipients acceptable in the field of skin care products, and the excipients comprise one or more of whitening agents, moisturizers, antioxidants, surfactants, preservatives, fragrances, and solvents.

9. The use according to claim 1, characterized in that The dosage form of the drug is ointment, patch, pill, tablet, powder, capsule, granule, liquid, injection, gel or suppository; The skin care product is a solution, suspension, emulsion, cream, ointment, gel, soap, oil, dry powder, liquid foundation, wet powder or spray.

10. A cream for repairing skin damage caused by UV radiation, characterized in that: The cream contains an active ingredient, which is selected from one or more of hederacoside C and its deglycosylated derivatives, and the mass proportion of the active ingredient in the cream is 5%-8%; The deglycosylated derivative of hederacoside C is a compound represented by Formula P3 and / or Formula P5:

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