Method and composition for preventing or delaying skin aging and maintaining or improving skin conditions

Abstract

A method for preventing or delaying skin aging and maintaining or improving skin conditions. The method comprises: administering to a subject a composition comprising alitretinoin or a pharmaceutically acceptable salt, hydrate, ester, analog or derivative thereof. The method can effectively relieve skin photoaging, reduce fine lines and wrinkles on the skin, improve skin elasticity, skin firmness and skin glossiness, and whiten the skin and the like.

Description

Methods and compositions for preventing or delaying skin aging and maintaining or improving skin condition Technical Field

[0001] The present invention belongs to the technical field of skin care products, and specifically relates to a method and composition for preventing or delaying skin aging and maintaining or improving skin condition. Background Art

[0002] The skin is the primary barrier that isolates our bodies from the external environment and protects us from environmental stresses. However, as we age, everyone's skin inevitably ages. While all organs of the human body degenerate with age, skin aging is most evident: sagging, dullness, and wrinkles. Generally, after the age of 20, the number of dermal fibroblasts gradually decreases, reducing their ability to synthesize collagen. Simultaneously, increased protease release increases collagen degradation. Although the content of elastic fibers remains unchanged, the elastic fiber network curls, loosens, and loses its elasticity. The elastic fiber network in the papillary dermis decreases, sometimes even disappears, causing epidermal sagging and the formation of fine wrinkles.

[0003] Skin aging is a complex biological phenomenon influenced by both intrinsic factors (such as genetics, cellular metabolism, endocrine function, and excessive oxidative stress) and extrinsic factors (such as gravity, chemicals, toxins, pollutants, ultraviolet (UV) radiation, and ionizing radiation). Furthermore, it is important to note that telomeres are closely linked to aging: shortened telomeres are associated with increased disease incidence and decreased survival. Therefore, telomeres are often considered a biomarker of cellular aging, acting as a "biological clock" that triggers aging. With the rapid development of society, environmental pollution has become increasingly prominent. Long-term exposure to polluted environments makes the skin vulnerable to damage. The fast-paced lifestyle and increasing stress, coupled with the effects of various factors such as ultraviolet light, age, medications, and sugar consumption, can shorten cellular telomerase activity at the microscopic level. This, in turn, can lead to various skin problems at the macroscopic level, including enlarged pores, decreased elasticity, dryness, roughness, sagging, dullness, and even skin health issues. While skin aging is inevitable, proper treatment can significantly improve or maintain healthy skin.

[0004] Therefore, maintaining and improving skin condition is increasingly popular among consumers, and skincare products are emerging in large numbers. However, existing skincare products are not very effective in improving or maintaining healthy skin, and they only treat the symptoms rather than the root cause. Alitretinoin (9-cis RA, CAS-5300-03-8) has multiple biological activities (such as anti-inflammatory activities). Research on the safety and efficacy of compositions containing alitretinoin in alleviating skin aging is of great significance.

[0005] Summary of the Invention

[0006] One aspect of the present invention relates to a method for preventing or delaying skin aging and maintaining or improving skin condition, comprising administering to a subject a composition containing alitretinoin or a pharmaceutically acceptable salt, hydrate, ester, analog or derivative thereof.

[0007] In some embodiments, an effective amount of a composition comprising alitretinoin or a pharmaceutically acceptable salt, hydrate, ester, analog, or derivative thereof is administered to a subject.

[0008] In some embodiments, preventing or delaying skin aging and maintaining or improving skin condition is one or more of: alleviating skin photoaging, reducing fine lines and wrinkles, improving skin elasticity, improving skin firmness, improving skin radiance, brightness or texture, changing skin thickness, improving skin dullness or uneven skin surface, maintaining skin hydration, whitening skin, and improving or evening skin tone. The skin can be any skin surface selected from the group consisting of the scalp, face, neck, shoulders, back, arms, legs, hands, feet, chest, abdomen, and buttocks.

[0009] In some embodiments, the composition of the present invention maintains or improves skin condition by inhibiting tyrosinase activity and / or lightening melanin and / or increasing epidermal thickness and / or delaying telomerase shortening.

[0010] In some embodiments, administration includes internal administration and external application. External application includes application, injection and the like.

[0011] In some embodiments, the composition containing alitretinoin or a pharmaceutically acceptable salt, hydrate, ester, analog or derivative thereof is formulated as a nutritional supplement, food, beverage, cosmetic or skin care product.

[0012] In some embodiments, the composition containing alitretinoin or a pharmaceutically acceptable salt, hydrate, ester, analog or derivative thereof is prepared as a solid preparation or a liquid preparation.

[0013] In some embodiments, the composition containing alitretinoin or a pharmaceutically acceptable salt, hydrate, ester, analog or derivative thereof is in the form of a suppository, tablet, pill, granule, powder, film, capsule, beverage, aerosol, elixir, tincture, tonic, liquid suspension, syrup, lotion, serum, emulsion, cream, gel, mask, or dressing.

[0014] In some embodiments, the composition comprising alitretinoin or a pharmaceutically acceptable salt, hydrate, ester, analog or derivative thereof is administered at a dose of 0.1 μM to 1 M or 0.1-500 mg or 0.001 to 50.0 weight % (w / w).

[0015] In some embodiments, the composition is administered at least once daily for 1 week to 7 weeks, 2 weeks to 7 weeks, 3 weeks to 7 weeks, or 4 weeks to 7 weeks.

[0016] In some embodiments, the present method includes oral administration, intravenous injection, intramuscular injection, intraperitoneal injection, local instillation, topical or sublingual administration, etc. For example, the composition is administered in a single dose or multiple divided doses, with a daily dose range of 0.1 mg to 2000 mg, 2 mg to 2000 mg, 5 mg to 1500 mg, 10 mg to 1200 mg, 20 mg to 1000 mg, 20 mg to 300 mg, 30 mg to 800 mg, 30 mg to 500 mg, 40 mg to 300 mg, 0.1 to 400 mg, 0.5 to 300 mg, 1 to 200 mg, 2 to 200 mg, 5 to 200 mg, 10 to 200 mg, 3 to 150 mg, 10 to 150 mg, 3 to 100 mg, 10 to 100 mg, 5 to 90 mg, 8 to 80 mg, 10 to 80 mg, 15 to 70 mg, 20 to 60 mg. In some embodiments, the daily dose range is 0.005% to 45%, 0.005% to 40%, 0.005% to 30%, 0.005% to 25%, 0.01% to 40%, 0.01% to 30%, 0.01% to 25%, 0.01% to 15%, 0.05% to 30%, 0.05% to 20%, 0.05% to 10%, 0.1% to 35%, 0.1% to 25%, 0.1% to 10%, 0.2% to 25%, 0.2% to 10%, 0.5% to 9%, 1% to 7%, 2% to 7%, 0.005% to 7%, 0.005% to 9%, 0.001% to 25%, 0.001% to 10%, 0.001% to 3%, 0.002% to 2%, 0.003% to 1%, 0.005% to 10%, 0.005% to 0.8%, 0.008% to 10%, 0.008% to 0.5%, 0.01% to 20%, 0.01% to 10%, 0.01% to 1%, 0.01% to 0.1%, 0.05% to 20%, 0.05% to 10%, 0.05% to 5% by weight (w / w). In some embodiments, the daily dose range is 0.1 μM to 1 M (e.g., 0.1 μM to 500 μM, 1 μM to 500 μM, 1 μM to 5 mM, 1 μM to 500 mM, 5 μM to 500 μM, 5 μM to 5 mM, 5 μM to 100 mM, 5 μM to 500 mM, 50 μM to 500 μM, 50 μM to 5 mM).

[0017] Another aspect of the present invention relates to the use of a composition containing alitretinoin or a pharmaceutically acceptable salt, hydrate, ester, analog, or derivative thereof in the manufacture of a nutritional supplement, food, beverage, cosmetic, or skin care product for preventing or delaying skin aging and maintaining or improving skin condition. Another aspect of the present invention relates to the use of alitretinoin or a pharmaceutically acceptable salt, hydrate, ester, analog, or derivative thereof in the manufacture of a composition for preventing or delaying skin aging and maintaining or improving skin condition.

[0018] In some embodiments, preventing or delaying skin aging and maintaining or improving skin condition is one or more of alleviating skin photoaging, reducing skin fine lines and wrinkles, improving skin elasticity, improving skin firmness, improving skin gloss, brightness or texture, changing skin thickness, improving skin dullness or uneven skin surface, keeping skin hydrated, whitening skin, and improving or evening out skin tone.

[0019] In some embodiments, the composition of the present invention maintains or improves skin condition by inhibiting tyrosinase activity and / or lightening melanin and / or increasing epidermal thickness and / or delaying telomerase shortening.

[0020] In some embodiments, administration includes internal administration and external application. External application includes application, injection and the like.

[0021] In some embodiments, the composition containing alitretinoin or a pharmaceutically acceptable salt, hydrate, ester, analog or derivative thereof is formulated as a nutritional supplement, food, beverage, cosmetic or skin care product.

[0022] In some embodiments, the composition containing alitretinoin or a pharmaceutically acceptable salt, hydrate, ester, analog or derivative thereof is prepared as a solid preparation or a liquid preparation.

[0023] In some embodiments, the composition containing alitretinoin or a pharmaceutically acceptable salt, hydrate, ester, analog or derivative thereof is in the form of a suppository, tablet, pill, granule, powder, film, capsule, beverage, aerosol, elixir, tincture, tonic, liquid suspension, syrup, lotion, serum, emulsion, cream, gel, mask, or dressing.

[0024] In some embodiments, the composition comprising alitretinoin or a pharmaceutically acceptable salt, hydrate, ester, analog or derivative thereof is administered at a dose of 0.1 μM to 1 M or 0.1-500 mg or 0.001 to 50.0 weight % (w / w).

[0025] In some embodiments, the composition is administered in a single dose or in divided doses with a daily dose range of 0.1 mg to 2000 mg, 2 mg to 2000 mg, 5 mg to 1500 mg, 10 mg to 1200 mg, 20 mg to 1000 mg, 20 mg to 300 mg, 30 mg to 800 mg, 30 mg to 500 mg, 40 mg to 300 mg, 0.1 to 400 mg, 0.5 to 300 mg, 1 to 200 mg, 2 to 200 mg, 5 to 200 mg, 10 to 200 mg, 3 to 150 mg, 10 to 150 mg, 3 to 100 mg, 10 to 100 mg, 5 to 90 mg, 8 to 80 mg, 10 to 80 mg, 15 to 70 mg, 20 to 60 mg. In some embodiments, the daily dose range is 0.005% to 45%, 0.005% to 40%, 0.005% to 30%, 0.005% to 25%, 0.01% to 40%, 0.01% to 30%, 0.01% to 25%, 0.01% to 15%, 0.05% to 30%, 0.05% to 20%, 0.05% to 10%, 0.1% to 35%, 0.1% to 25%, 0.1% to 10%, 0.2% to 25%, 0.2% to 10%, 0.5% to 9%, 1% to 7%, 2% to 7%, 0.005% to 7%, 0.005% to 9%, 0.001% to 25%, 0.001% to 10%, 0.001% to 3%, 0.002% to 2%, 0.003% to 1%, 0.005% to 10%, 0.005% to 0.8%, 0.008% to 10%, 0.008% to 0.5%, 0.01% to 20%, 0.01% to 10%, 0.01% to 1%, 0.01% to 0.1%, 0.05% to 20%, 0.05% to 10%, 0.05% to 5% by weight (w / w). In some embodiments, the daily dose range is 0.1 μM to 1 M (e.g., 0.1 μM to 500 μM, 1 μM to 500 μM, 1 μM to 5 mM, 1 μM to 500 mM, 5 μM to 500 μM, 5 μM to 5 mM, 5 μM to 100 mM, 5 μM to 500 mM, 50 μM to 500 μM, 50 μM to 5 mM).

[0026] Another aspect of the present invention relates to a skin care product or cosmetic comprising a composition of alitretinoin or a pharmaceutically acceptable salt, hydrate, ester, analog or derivative thereof, which is used for maintaining or improving skin conditions.

[0027] In some embodiments, the skin care product or cosmetic maintains or improves skin condition by inhibiting tyrosinase activity and / or lightening melanin and / or increasing epidermal thickness and / or delaying telomerase shortening.

[0028] In some embodiments, the skin care product or cosmetic is in the form of a suppository, tablet, pill, granule, powder, film, capsule, beverage, aerosol, spirit, tincture, tonic, liquid suspension, syrup, lotion, serum, emulsion, cream, gel, mask, or dressing.

[0029] In some embodiments, the amount of the composition comprising alitretinoin or a pharmaceutically acceptable salt, hydrate, ester, analog or derivative thereof in the skin care product or cosmetic is 0.1 μM to 1 M or 0.1-500 mg or 0.001 to 50.0 weight % (w / w).

[0030] In some embodiments, the skin care product or cosmetic is administered in a single dose or multiple divided doses, with a daily dose range of 0.1 mg to 2000 mg, 2 mg to 2000 mg, 5 mg to 1500 mg, 10 mg to 1200 mg, 20 mg to 1000 mg, 20 mg to 300 mg, 30 mg to 800 mg, 30 mg to 500 mg, 40 mg to 300 mg, 0.1 to 400 mg, 0.5 to 300 mg, 1 to 200 mg, 2 to 200 mg, 5 to 200 mg, 10 to 200 mg, 3 to 150 mg, 10 to 150 mg, 3 to 100 mg, 10 to 100 mg, 5 to 90 mg, 8 to 80 mg, 10 to 80 mg, 15 to 70 mg, 20 to 60 mg. In some embodiments, the daily dose range is 0.005% to 45%, 0.005% to 40%, 0.005% to 30%, 0.005% to 25%, 0.01% to 40%, 0.01% to 30%, 0.01% to 25%, 0.01% to 15%, 0.05% to 30%, 0.05% to 20%, 0.05% to 10%, 0.1% to 35%, 0.1% to 25%, 0.1% to 10%, % to 20%, 0.01% to 10%, 0.01% to 1%, 0.01% to 0.1%, 0.05% to 20%, 0.05% to 10%, 0.05% to 5% by weight (w / w). In some embodiments, the daily dose range is 0.1 μM to 1 M (e.g., 0.1 μM to 500 μM, 1 μM to 500 μM, 1 μM to 5 mM, 1 μM to 500 mM, 5 μM to 500 μM, 5 μM to 5 mM, 5 μM to 100 mM, 5 μM to 500 mM, 50 μM to 500 μM, 50 μM to 5 mM).

[0031] The method of the present invention of administering a composition containing alitretinoin or a pharmaceutically acceptable salt, hydrate, ester, analog or derivative thereof to a subject can maintain or improve skin conditions, including alleviating skin photoaging, reducing skin fine lines and wrinkles, improving skin elasticity, improving skin firmness, improving skin gloss, brightness or texture, changing skin thickness, improving skin dullness or uneven skin surface, keeping skin hydrated, whitening skin, and improving or evening skin tone. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] FIG1 is a graph showing the effects of different concentrations of alitretinoin on cell viability.

[0033] Figure 2 is a graph showing the inhibitory effect of different concentrations of alitretinoin on cellular melanin secretion.

[0034] FIG3 shows the effect of alitretinoin (9-cis RA) of the present invention on skin essential aging after 9 months of use.

[0035] FIG4 shows the results of the percentage of short telomeres in cells treated with alitretinoin (9-cis RA) of the present invention.

[0036] FIG5 shows the results of telomere shortening rate of cells treated with alitretinoin (9-cis RA) of the present invention. DETAILED DESCRIPTION

[0037] Although the present invention will be described in conjunction with the embodiments, it should be understood that they are not intended to limit the present invention to these embodiments. On the contrary, the present invention is intended to cover substitutions, modifications and equivalents, which may be included within the spirit and scope of the present invention as defined in the claims. In addition, in the detailed description of the present invention, many specific details are set forth in order to provide a thorough understanding of the present invention. However, it is apparent to one of ordinary skill in the art that the present invention can be practiced without these specific details. In other cases, well-known methods, procedures, components and other features are not described in detail.

[0038] As used herein, the term "or" is intended to include "and" and "or." In other words, the term "or" can also be replaced with "and / or."

[0039] As used herein, the singular forms "a," "an," and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise.

[0040] As used herein, the term "comprises" or "includes" or variations thereof refers to instances where the term is used in its non-limiting sense, meaning that items following the term are included, but items not specifically mentioned are not excluded. It also includes the more restrictive verbs 'consisting essentially of' and 'consisting of.'

[0041] As used herein, the terms "subject" or "individual" are used interchangeably to refer to any subject to which the methods and compositions of the present disclosure can be applied or administered. The subject may have a disease or condition, but the subject does not need to be ill to benefit from the methods and compositions of the present disclosure. Any subject can take the disclosed compositions or become a recipient of the disclosed methods. Herein, the term "subject" refers to an animal (e.g., birds, reptiles, and mammals). In certain embodiments, the subject can be a mammal including non-primates (e.g., camels, donkeys, zebras, cows, horses, cats, dogs, rats, and mice) and primates (e.g., monkeys, chimpanzees, humans). In certain embodiments, the subject can be a non-human mammal. In other embodiments, the subject can be a human.

[0042] As used herein, the term "administering" refers to directly administering a compound or a pharmaceutically acceptable salt or composition of a compound to a subject, or administering a prodrug derivative or analog of a compound or a pharmaceutically acceptable salt or composition of a compound to a subject, which can form an equivalent amount of an active compound in the subject's body.

[0043] The method of expectation comprises the alitretinoin or alitretinoin substitute of at least 1mg being used every day, and this depends on the specific preparation and form containing alitretinoin.This amount every day can be used once or repeatedly.Typically, the effective amount of alitretinoin or alitretinoin derivative in the composition containing alitretinoin used once or repeatedly every day is 1 milligram (mg) to 2000mg or between 1 milligram (mg) to 2000mg.One or more dosages can be applied once a day in any time period.For example, effective dose can be applied every day, continue one day, several days, many days or apply every day indefinitely, alitretinoin can be applied for 7 days and above in a cycle, depends on the effect of hope. More typically, the amount of alitretinoin or alitretinoin derivatives used every day is 0.1 mg, 0.5 mg, 1 mg, 2 mg, 5 mg, 10 mg, 25 mg, 25 mg, 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 700 mg, 800 mg, 900 mg, 1000 mg, 1100 mg, 1200 mg, 1300 mg, 1400 mg, 1500 mg. More typically, the amount of alitretinoin or alitretinoin derivatives used every day is 1 mg to 1000 mg. Most typically, the amount of alitretinoin or alitretinoin derivatives used every day is 10-50 mg.

[0044] The effective amount of alitretinoin or alitretinoin derivatives in the alitretinoin-containing composition administered once or more daily is 0.001 wt % to 50.0 wt % or between 0.001 and 50.0 wt % (w / w). One or more doses can be administered once daily in any time period. For example, an effective dose can be administered daily for one day, several days, multiple days, or indefinitely, and alitretinoin can be administered for 7 days and above in a cycle, depending on the desired effect. More typically, the amount of alitretinoin or an alitretinoin derivative administered per day is 0.005%, 0.01%, 0.05%, 0.1%, 0.2%, 0.5%, 1%, 2%, 3%, 4%, 5%, 7%, 9%, 10%, 12%, 15%, 16%, 18%, 20%, 25%, 30%, 35%, 40%, 45%, 50% by weight (w / w). The amount of alitretinoin or alitretinoin derivatives administered once or multiple times per day can be 0.1 μM to 1M (e.g., 0.1 μM to 500 μM, 1 μM to 500 μM, 1 μM to 5mM, 1 μM to 100 mM, 1 μM to 500 mM, 5 μM to 500 μM, 5 μM to 5mM, 5 μM to 100 mM, 5 μM to 500 mM, 50 μM to 500 μM, 50 μM to 5mM). One dosage or multiple dosages can be administered in any time period of each day. For example, an effective dose can be given every day for one day, several days, many days, or indefinitely based on each day.

[0045] Various forms and formulations of compositions containing alitretinoin are contemplated. The composition will be formulated into a nutritional supplement or dietary supplement, (medical) food, skin care product, in liquid or solid form, a composition comprising alitretinoin, and may optionally include a nutritionally or pharmaceutically acceptable carrier. For oral administration, the composition of the present invention may be in any suitable form, including a solution, tablet, gel capsule, capsule or alternative nutritional food or nutritional supplement form. For topical application, the composition will be applied to the skin part to be treated, particularly the face, neck or hands, once or several times a day; the form of the composition includes lotion, skin softener, toner, skin cream, sunscreen, essence, facial mask, facial mask sheet, soap, shampoo, cleansing foam, cleansing lotion, cleansing cream, ointment, gel and massage cream, etc.

[0046] The technical features, implementation methods and beneficial effects of the present invention are further described in detail below with reference to specific implementation cases. Unless otherwise specified, the materials and reagents described in the following examples are all common commercial products and can be purchased on the market.

[0047] Example

[0048] Example 1 Whitening Test - Testing the Inhibitory Effect of the Present Invention on Tyrosinase

[0049] Materials and instruments:

[0050] Reagents: tyrosinase, tyrosine, potassium chloride, sodium chloride, potassium dihydrogen phosphate, sodium hydrogen phosphate, alitretinoin (9-cis RA), deionized water, dimethyl sulfoxide (DMSO).

[0051] Main instruments: magnetic stirrer, microplate reader, water purifier.

[0052] Experimental methods:

[0053] Solution Preparation: a) Phosphate Buffered Saline (PBS): 0.2g potassium chloride, 8g sodium chloride, 0.27g potassium dihydrogen phosphate, 3.58g sodium dihydrogen phosphate. First, add 800mL of distilled water and stir with a magnetic stirrer until completely dissolved. Then, adjust the pH to 6.8 and make up the volume to 1000mL. After autoclaving, store at 4°C or at room temperature. b) 2mM L-Tyrosine Solution: Weigh 36mg of L-tyrosine and add 100mL of water. Stir on a magnetic stirrer until completely dissolved to obtain the L-tyrosine solution. c) Tyrosinase Solution: Dissolve in phosphate buffered saline (PBS) to a final enzyme activity concentration of 250U / mL. d) Preparation of the Alitretinoin Test Solution: Dissolve 10mg of alitretinoin in 10mL of DMSO to obtain a 1mg / mL sample solution.

[0054] Tyrosinase inhibition assay: Add tyrosine solution and sample solution to a 96-well plate in the order shown in the table below and incubate at 30°C for 10 minutes. Then, add tyrosinase solution and mix thoroughly. Incubate at 30°C for another 20 minutes and measure absorbance at 475 nm.

[0055] The sample group was zeroed using blank control group 1, and the blank control group was zeroed using blank control group 3. The absorbance of the sample group was A, the absorbance of blank control group 1 was A1, the absorbance of blank control group 2 was A2, and the absorbance of blank control group 3 was A3. The inhibition rate was calculated as follows: Inhibition rate = [1 - [(A - A1) / (A2 - A3)] × 100%.

[0056] The results showed that the average inhibition rate of tyrosinase activity by alitretinoin was 90%.

[0057] Example 2 Whitening Test - Testing the Melanin Lightening Effect of the Present Invention

[0058] Materials and instruments:

[0059] Materials: Alitretinoin (9-cis RA), α-melanocyte-stimulating hormone (α-MSH), B16-F1 cells, MTT kit, DMEM medium, fetal bovine serum, penicillin-streptomycin double antibody, cell culture flasks, 96-well plates, and 6-well plates. All other reagents were commercially available and analytical grade.

[0060] Instruments: microplate reader, cell culture incubator.

[0061] Cell culture

[0062] Mouse melanoma cells (B16-F1) were cultured in Dulbecco's modified Eagle's medium (DMEM) containing 1% penicillin-streptomycin and 10% fetal bovine serum in a 5% CO2 incubator at 37°C.

[0063] Cell viability (MTT method)

[0064] The MTT assay was used to evaluate the effect of alitretinoin (9-cis RA) on cell viability. Briefly, cells (0.5×10 5 Cells were seeded into 96-well plates (100 cells / well) and treated with 9-cis RA (0, 0.5, 1, 2, and 4 μg / mL) for 72 h. MTT reduction was quantified by dissolving the formazine precipitate in dimethyl sulfoxide (DMSO) and measuring absorbance at 570 nm using a microplate reader.

[0065] Figure 1 shows the effect of different concentrations of alitretinoin on cell viability. As can be seen from the figure, there is no decrease in cell viability at each concentration, indicating that the two have no toxic effects on cells within this concentration range.

[0066] Melanin content detection

[0067] B16-F1 cells were grown in 6-well plates at a density of 0.5 × 10 5 Cells were cultured overnight at a density of 100 cells / well and then treated with a non-toxic concentration of 9-cis RA (based on cell viability results) in DMEM containing α-MSH (10 nM). The cells were incubated at 37°C in 5% CO2 for 73 h. The culture supernatant was collected and centrifuged at 16,000 × g for 4 min. Melanin content was determined based on A490 values.

[0068] Figure 2 shows the inhibitory effect of different concentrations of alitretinoin on cell melanin secretion. It can be seen from the figure that alitretinoin at different concentrations has an inhibitory effect on the production of melanin.

[0069] Example 3 Anti-aging test - Testing the effect of the present invention on increasing epidermal thickness, reducing wrinkles, and improving skin tone

[0070] Six women aged 68 to 79 years (mean, 74 years) applied 0.025% alitretinoin (9-cis RA) cream to one inner thigh and a non-9-cis RA cream to the other thigh (nose) once daily for 9 months. The participants were healthy and not taking medications for age-related conditions. Four-millimeter needle biopsies were obtained before and after 9 months of treatment. Initial appearance was mild laxity, slight dryness, and a lighter tone.

[0071] By visual observation, compared with the blank control, the side treated with 9-cis RA showed significant improvement, with reduced skin wrinkles, firmness, and a pinkish hue.

[0072] Figure 3 shows the effect of 9-cis RA (9-cis RA) on intrinsic skin aging after 9 months of use. As shown in Figure 3, the average viable epidermal thickness increased from 60 μm in the blank group to approximately 135 μm after 9-cis RA application.

[0073] Example 4 Anti-aging test - testing the present invention to delay the shortening of telomerase and even extend the length of telomerase

[0074] Primary cultures of human neonatal dermal fibroblasts (with a lifespan of approximately 2 months) were established under standard culture conditions. These cells were chosen because primary cell lines are needed to observe the occurrence of normal cell aging and telomere shortening. Cells were cultured at 5×10 3 cells / cm 2 Cultivate and grow in fibroblast culture medium.

[0075] Cell viability:

[0076] MTT assay: The MTT assay is a colorimetric method that uses a yellow tetrazolium salt (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, or MTT) to measure the metabolic activity of cells as an indicator of cell viability. The MTT reduction rate is an indicator of mitochondrial functional integrity. To determine the cytotoxicity of alitretinoin (9-cis RA) on primary neonatal fibroblasts, cells were incubated in 96-well plates at 0.35 × 10 4 cells / plate and 0.5×10 4Cells / plate, 72h and 1 week. After 24h of incubation, the cells were washed once with phosphate-buffered saline (PBS) and treated with the corresponding compound in cell culture medium. Alitretinoin (9-cis RA) was dissolved in dimethyl sulfoxide (DMSO). Serial half dilutions were used (final concentrations of 0.04, 0.1, 0.3 and 1.0 mg / ml). During the experiment, each condition was tested three times with and without 10μM H2O2 in the culture medium. Because it was observed that 10μM H2O2 causes oxidative damage to cells, maintains cell proliferation, and does not cause cell death during the 2-month expansion period used in the study. Positive and negative controls used 8mM methyl methane sulfonate (MMS) and 100% DMSO, respectively. After re-addition, the cells were incubated for 72h and 1 week, respectively. The culture medium containing the compound was replaced every two days. After the treatment period, wash the cells twice with PBS and replace the culture medium with 0.5 mg / ml MTT reagent in Dulbecco's Modified Eagle Medium (DMEM) without phenol red. Gently shake the culture dish and incubate for 4 hours. After incubation, remove the culture medium and replace it with DMSO. Gently shake the dish to dissolve the formaldehyde crystals. Measure the absorbance at a wavelength of 570 nm using a reader.

[0077] The percentage of cell death was calculated as follows: Cell death (%) = OD sample -OD PC =OD NC -OD PC *100. Among them, OD NC Optical density of negative control, OD PC is the optical density of the positive control.

[0078] All 9-cis RA concentrations (0.04, 0.1, 0.3, and 1.0 mg / ml) produced <20% cell death, indicating no significant toxic effects. That is, 9-cis RA concentrations up to 1.0 mg / ml did not produce any deleterious effects.

[0079] Measuring telomere length

[0080] All telomere length measurements were performed using telomere analysis technology. Telomere length was measured using high-throughput (HT) Q-FISH (quantitative fluorescence in situ hybridization) technology. The percentage of short telomeres and the rate of telomere shortening were measured and calculated for the DMSO group (blank group) and the alitretinoin (9-cis RA) group (final concentrations of 0.04, 0.1, 0.3, and 1.0 mg / ml, respectively).

[0081] Compared with the blank group, the percentage of short telomeres (<3 kbp or <3000 bp) in cells treated with 9-cis RA (especially 1.0 mg / ml) at week 4 was significantly reduced, indicating that telomerase shortening was delayed, and the effect was particularly obvious with increasing concentration (Figure 4).

[0082] As shown in Figure 5, compared with the blank control group, the telomere shortening rate of cells treated with 9-cis RA at week 4 significantly decreased with increasing concentrations. More interestingly, the telomere shortening rate of cells treated with 0.3 and 1.0 mg / ml 9-cis RA was negative, which seems to indicate telomere lengthening.

[0083] Briefly, telomeres are hybridized with a fluorescent peptide nucleic acid probe (PNA) that recognizes three telomeric repeats (sequence: Alexa488-OO-CCCTAACCCTAACCCTAA). The intensity of the fluorescent signal from the telomeric PNA probe hybridized to a given telomere is proportional to the length of the telomere. The fluorescence intensity is translated into base pairs (bp) using a standard regression curve generated using control cell lines with known telomere lengths. Not only does it measure telomere length in absolute bp units, it also provides an assessment of the distribution of telomere lengths, the 50th and 20th percentiles, and the percentage of short telomeres, allowing for a more comprehensive analysis of each sample.

[0084] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Any person skilled in the art may make various changes, modifications, substitutions and variations to these embodiments without departing from the principles and purpose of the present invention. The scope of the present invention is defined by the claims and their equivalents.

Claims

1. A method for preventing or delaying skin aging and maintaining or improving skin condition, characterized in that: The method comprises administering to a subject a composition comprising alitretinoin or a pharmaceutically acceptable salt, hydrate, ester, analog or derivative thereof.

2. The method according to claim 1, characterized in that The prevention or delay of skin aging and the maintenance or improvement of skin condition are one or more of alleviating skin photoaging, reducing skin fine lines and wrinkles, improving skin elasticity, improving skin firmness, improving skin gloss, brightness or texture, changing skin thickness, improving skin dullness or uneven skin surface, keeping skin moisturized, whitening skin, improving or even skin tone.

3. The method according to claim 1 or 2, characterized in that: The composition prevents or delays skin aging and maintains or improves skin condition by inhibiting the activity of tyrosinase or / and lightening melanin or / and increasing epidermal thickness or / and delaying the shortening of telomerase.

4. The method according to any one of claims 1 to 3, characterized in that The administration includes internal administration and external application.

5. The method according to any one of claims 1 to 4, characterized in that The composition containing alitretinoin or a pharmaceutically acceptable salt, hydrate, ester, analog or derivative thereof is formulated into a nutritional supplement, food, beverage, cosmetic or skin care product.

6. The method according to any one of claims 1 to 5, characterized in that The composition containing alitretinoin or a pharmaceutically acceptable salt, hydrate, ester, analog or derivative thereof is prepared as a solid preparation or a liquid preparation.

7. The method according to any one of claims 1 to 6, characterized in that The composition containing alitretinoin or its pharmaceutically acceptable salt, hydrate, ester, analog or derivative is in the form of suppository, tablet, pill, granule, powder, film, capsule, beverage, aerosol, spirit, tincture, tonic, liquid suspension, syrup, lotion, essence, emulsion, cream, gel, facial mask or dressing.

8. The method according to any one of claims 1 to 7, characterized in that The composition containing alitretinoin or a pharmaceutically acceptable salt, hydrate, ester, analog or derivative thereof is administered at a dosage of 0.1 μM to 1 M or 0.1-500 mg or 0.001 to 50.0 weight % (w / w).

9. The method according to any one of claims 1 to 8, characterized in that The composition containing alitretinoin or a pharmaceutically acceptable salt, hydrate, ester, analog or derivative thereof is administered at a dosage of 1 μM to 500 mM or 20 to 300 mg or 0.005 to 9 weight % (w / w).

10. Use of alitretinoin or a pharmaceutically acceptable salt, hydrate, ester, analog or derivative thereof in the manufacture of a composition for preventing or delaying skin aging and maintaining or improving skin condition, characterized in that: The composition comprises an effective amount of alitretinoin or a pharmaceutically acceptable salt, hydrate, ester, analog or derivative thereof.

11. The use according to claim 10, characterized in that The prevention or delay of skin aging and maintenance or improvement of skin condition is one or more of alleviating skin photoaging, reducing skin fine lines and wrinkles, improving skin elasticity, improving skin firmness, improving skin gloss, brightness or texture, changing skin thickness, improving skin dullness or uneven skin surface, keeping skin moisturized, whitening skin, improving or even skin tone.

12. The use according to claim 10 or 11, characterized in that The composition prevents or delays skin aging and maintains or improves skin condition by inhibiting the activity of tyrosinase or / and lightening melanin or / and increasing epidermal thickness or / and delaying the shortening of telomerase.

13. The use according to any one of claims 10 to 12, characterized in that The administration includes internal administration and external application.

14. The use according to any one of claims 10 to 13, characterized in that The composition containing alitretinoin or a pharmaceutically acceptable salt, hydrate, ester, analog or derivative thereof is formulated into a nutritional supplement, food, beverage, cosmetic or skin care product.

15. The use according to any one of claims 10 to 14, characterized in that The composition containing alitretinoin or a pharmaceutically acceptable salt, hydrate, ester, analog or derivative thereof is prepared as a solid preparation or a liquid preparation.

16. The use according to any one of claims 10 to 15, characterized in that The composition containing alitretinoin or its pharmaceutically acceptable salt, hydrate, ester, analog or derivative is in the form of suppository, tablet, pill, granule, powder, film, capsule, beverage, aerosol, spirit, tincture, tonic, liquid suspension, syrup, lotion, essence, emulsion, cream, gel, facial mask or dressing.

17. The use according to any one of claims 10 to 16, characterized in that The composition containing alitretinoin or a pharmaceutically acceptable salt, hydrate, ester, analog or derivative thereof is administered at a dosage of 0.1 μM to 1 M or 0.1-500 mg or 0.001 to 50.0 weight % (w / w).

18. The use according to any one of claims 10 to 17, characterized in that The composition containing alitretinoin or a pharmaceutically acceptable salt, hydrate, ester, analog or derivative thereof is administered at a dosage of 1 μM to 500 mM or 20 to 300 mg or 0.005 to 9 weight % (w / w).

19. A skin care product or cosmetic comprising a composition of alitretinoin or a pharmaceutically acceptable salt, hydrate, ester, analog or derivative thereof, characterized in that: The skin care products or cosmetics are used to prevent or delay skin aging and maintain or improve skin conditions.

20. The skin care product or cosmetic according to claim 19, characterized in that: The skin care product or cosmetics prevents or delays skin aging and maintains or improves skin condition by inhibiting the activity of tyrosinase or / and lightening melanin or / and increasing epidermal thickness or / and delaying the shortening of telomerase.

21. The skin care product or cosmetic according to claim 19 or 20, characterized in that: The skin care products or cosmetics are in the form of suppositories, tablets, pills, granules, powders, films, capsules, beverages, aerosols, spirits, tinctures, tonics, liquid suspensions, syrups, lotions, essences, emulsions, creams, gels, facial masks, and dressings.

22. The skin care product or cosmetic according to any one of claims 19 to 21, characterized in that: The amount of the composition comprising alitretinoin or a pharmaceutically acceptable salt, hydrate, ester, analog or derivative thereof in the skin care product or cosmetic is 0.1 μM to 1 M or 0.1-500 mg or 0.001 to 50.0 weight % (w / w).

23. The skin care product or cosmetic according to any one of claims 19 to 22, characterized in that: The amount of the composition comprising alitretinoin or a pharmaceutically acceptable salt, hydrate, ester, analog or derivative thereof in the skin care product or cosmetic is 1 μM to 500 mM or 20 to 300 mg or 0.005 to 9 weight % (w / w).

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