KR101882923B1 - Cosmetic composition for protecting skin and inhibiting damage of skin against ultraviolet comprising S-Methylmethionine sulfonium derivative - Google Patents

Abstract

The present invention relates to a composition for inhibiting skin damage and protecting skin caused by ultraviolet rays, which comprises a derivative of S-methylmethionine sulfonium (SMMS) or a pharmaceutically acceptable salt thereof as an active ingredient, Provides cosmetic compositions, quasi-drugs and pharmaceutical compositions for skin damage prevention and skin protection by ultraviolet rays.

Description

[0001] The present invention relates to a cosmetic composition for protecting skin and protecting skin from ultraviolet rays, which contains a derivative of S-methylmethionine sulfonium as an active ingredient,

The present invention relates to a composition for inhibiting skin damage and protecting skin caused by ultraviolet rays, which comprises a derivative of S-methylmethionine sulfonium (SMMS) or a pharmaceutically acceptable salt thereof as an active ingredient, Provides cosmetic compositions, quasi-drugs and pharmaceutical compositions for skin damage prevention and skin protection by ultraviolet rays.

Stimuli such as externally applied ultraviolet rays, environmental pollutants, rapid temperature changes, etc., induce a variety of biochemical changes in skin cells, thereby causing a cell damaging reaction. The double ultraviolet rays are in contact with the skin to increase DNA damage and / or active oxygen production, thereby increasing cell denaturation or apoptosis.

The ultraviolet rays irradiated to the skin pass through one of four routes: direct reflection at the boundary of the skin layer (that is, epidermal reflection, dermal reflection), scattering in the skin tissue, collision with fibers or particles, . Dual absorbed UV rays cause photochemical reactions in the skin, resulting in skin lesions. The penetration degree of the skin differs according to the wavelength. UVA of long wavelength penetrates deeply into the dermis. However, UVB of 315-280 nm and UVC of 280-100 nm have a relatively low penetration degree because of a short wavelength. Thus, UVA can cause severe dermal reactions, and UVB can cause severe skin reactions. UVA is mainly a skin tanning reaction, UVB is the main cause of sunburn and promotes pigmentation.

S-Methylmethionine sulfonium (SMMS) is a methionine derivative and is an active substance abundantly contained in cabbage, cola, turnip, tomato, celery and the like. SMMS has a therapeutic effect to protect gastrointestinal ulceration and is widely known as vitamin U. For example, it has been reported that SMMS can have a rapid therapeutic effect on peptic ulcer. In addition, it has been known that SMMS can exhibit a wound healing effect and a light protection effect on skin, and thus can be used as a raw material for a cosmetic composition. For example, animal studies have shown that topical administration of SMMS for both trauma and chemical wounds can facilitate suturing of the wound and re-epithelialization of the wound.

In addition, treatment of SMMS can promote not only the growth of human dermal fibroblast (hDF) but also migration, and because SMMS activates the ERK1 / 2 pathway, it promotes hDF proliferation and promotes migration . In addition, SMMS can protect skin cells in the UVB irradiation environment by eliminating reactive oxygen species (ROS). Treatment of SMMS can reduce UVB-induced erythema index and Langerhans cell deficiency.

However, despite the fact that SMMS can effectively exhibit skin regeneration effect, it has a disadvantage that stability is not maintained, and thus, there is a limitation in use as a drug. For example, it has been reported that the content of SMMS is reduced to 62% in cabbage that is not under temperature control for storage. Another disadvantage is that the odor is generated from the sulfonium functional group of SMMS. For example, it is known that in the heating step during the melting process for formulating SMMS, odor may occur because the SMMS structure breaks down to form dimethyl sulfide. Accordingly, when SMMS is used in a cosmetic composition, it has a problem that the smell is not good, and it is not stable and thus it can not be used industrially.

As a result, SMMS has a disadvantage that it can not be used as a cosmetic composition even though it has excellent wound restorative effect and skin protection against UVB. Therefore, a need has arisen to present, as an alternative, a novel compound which is more stable among the derivatives having a structure similar to that of SMMS conventionally used and does not generate malodor.

Accordingly, the present inventors have made efforts to select a novel compound capable of exhibiting a skin protection effect by ultraviolet rays. As a result, it has been found that (2S, 4S) and (2R, 4S) -2-phenylthiazolidine- (2S, 4S), (2R, 4S) -2-phenylthiazolidine-4-carboxylic acid, Compound 15) and (2S, 4R) (2S, 4S) -2-phenyl-1,3-thiazin-4-carboxylic acid ((2S, 4R) thiazinane-4-carboxylic acid (Compound 16) promotes cell proliferation not only against human dermal fibroblast (hDF) and dermal keratinocyte but also against cytoprotective effect against ultraviolet (UVB) The present invention has been completed.

(2S, 4S), (2S, 4S) -2-phenylthiazolidine-4-carboxylic acid (Compound 15) and (2S, 4R) , (2R, 4R), and (2R, 4S) -2-phenyl-1,3-thiazin-4-carboxylic acid (Compound 16) in skin fibroblasts and skin keratinocytes And the present invention has been completed.

Accordingly, an object of the present invention is to provide a cosmetic composition for skin damage prevention and skin protection by ultraviolet rays.

It is also an object of the present invention to provide a pharmaceutical composition for preventing or treating skin damage caused by ultraviolet rays.

In order to achieve the above object, the present invention provides a method for inhibiting skin damage and protecting skin caused by ultraviolet rays, comprising a derivative of S-methylmethionine sulfonium (SMMS) or a pharmaceutically acceptable salt thereof as an active ingredient Thereby providing a cosmetic composition.

The present invention also provides a pharmaceutical composition for preventing or treating skin damage caused by ultraviolet rays, which comprises a derivative of S-methylmethionine sulfonium or a pharmaceutically acceptable salt thereof as an active ingredient.

In a preferred embodiment of the present invention, the derivative of S-methylmethionine sulfonium can be a compound having the structure of the following formula (1)

[Chemical Formula 1]

.

.

(In the above formula, R ₁ is a heterocycle of 5 or 6 atoms, including either or both of S and N.)

In one preferred embodiment of the present invention, the compound is (2S, 4S) and (2R, 4S) -2-phenylthiazolidine-4-carboxylic acid represented by the following formula (2S, 4R), (2S, 4S), (2R, 4R), (2R, 4S) -2-phenyl-1,3- thiazin-

(2)

; 및

; And

(3)

.

.

In another preferred embodiment of the present invention, the ultraviolet ray may be UVB.

In another preferred embodiment of the present invention, the skin damage caused by ultraviolet rays may be any one of an image, erythema, or pigment deposition which is generated in the epidermis or dermis by ultraviolet rays.

Accordingly, the present invention provides a cosmetic composition for preventing skin damage and protecting skin caused by ultraviolet rays, which comprises a derivative of SMMS or a pharmaceutically acceptable salt thereof as an active ingredient.

(2S, 4S), (2R, 4S) -2-phenylthiazolidine-4-carboxylic acid (Compound 15) and (2S, 4R) , 4R) and (2R, 4S) -2-phenyl-1,3-thiazin-4-carboxylic acid (Compound 16) exhibit cell proliferation promoting effect in human dermal fibroblast and skin keratinocyte, And is effective as an active ingredient of cosmetic compositions, quasi-drugs, and pharmaceutical compositions for skin damaging and skin protection by ultraviolet rays.

In addition, the compound 15 and the compound 16 of the present invention can overcome the limitations of instability and odor of S-methylmethionine sulfonium (SMMS) which is conventionally used in the skin damaging and skin protecting composition by ultraviolet rays, to be.

Figure 1 shows the promoting effect of Compound 15 and 16 of the present invention on dermal fibroblast proliferation and cytoprotective effect against UVB :
FIG. 1A is a graph showing the promoting effect of various SMMS derivatives on skin fibroblast proliferation; And
Figure 1B shows the cytoprotective effect of various SMMS derivatives against UVB.
Figure 2 shows the synthesis steps of compounds 15 and 16 of the present invention:
2A and 2B are diagrams showing the structures of Compound 15 and Compound 16 of the present invention, respectively; And
FIG. 2C is a diagram showing the synthesis process of the compound 15 and the compound 16 of the present invention. FIG.
Figure 3 shows the skin protection effect of Compound 15 and Compound 16 against dermal fibroblast (hDF)
FIG. 3A is a graph showing the cell proliferation promoting effect of Compound 15 on hDF; FIG.
3B is a graph showing the cell proliferation promoting effect of Compound 16 on hDF;
Figure 3c is a graph showing the cytoprotective effect of compound 15 on hDF irradiated with UVB;
Figure 3d is a diagram showing the cytoprotective effect of Compound 16 against hDF irradiated with UVB;
FIG. 3E shows the control of mRNA expression levels of collagen synthase and melanin synthase for 15 and 16 against hDF irradiated with UVB; FIG. And
Figure 3f shows the effect of ROS elimination on Compound 15 and Compound 16 against hDF irradiated with UVB.
Figure 4 shows the skin protection effect of Compound 15 and Compound 16 on HaCaT cells which are dermal keratinocytes:
4A is a graph showing the cell proliferation promoting effect of Compound 15 on HaCaT cells;
4B is a graph showing the cell proliferation promoting effect of Compound 16 on HaCaT cells;
Fig. 4C is a chart showing the cytoprotective effect of Compound 15 on HaCaT cells irradiated with UVB; Fig.
Figure 4d is a diagram showing the cytoprotective effect of Compound 16 on HaCaT cells irradiated with UVB; And
Fig. 4E shows the effect of ROS elimination on Compound 15 and Compound 16 against HaCaT cells irradiated with UVB. Fig.

Hereinafter, the present invention will be described in detail.

As described above, conventional S-methylmethionine sulfonium (SMMS) has been widely used as a compound capable of inhibiting skin damage due to ultraviolet rays, but since the structure of the compound is not stable, production of a drug is not easy, So that it is limited to use as a component of a cosmetic composition.

(2S, 4S), (2R, 4S) -2-phenylthiazolidine-4-carboxylic acid (Compound 15) and (2S, 4R) , 4R) and (2R, 4S) -2-phenyl-1,3-thiazin-4-carboxylic acid (Compound 16) exhibit cell proliferation promoting effect in human dermal fibroblast and skin keratinocyte, And is effective as an active ingredient of cosmetic compositions, quasi-drugs, and pharmaceutical compositions for skin damaging and skin protection by ultraviolet rays. In addition, the compound 15 and the compound 16 of the present invention can overcome the limitations of instability and odor of S-methylmethionine sulfonium (SMMS) which is conventionally used in the skin damaging and skin protecting composition by ultraviolet rays, to be.

Accordingly, the present invention provides a cosmetic composition for inhibiting skin damage and skin protection by ultraviolet rays, which comprises a derivative of S-methylmethionine sulfonium or a pharmaceutically acceptable salt thereof as an active ingredient.

The "S-methylmethionine sulfonium derivative" of the present invention is preferably a compound having a structure represented by the following formula (1), wherein R ₁ represents any one or both of S and N , 5 or 6 atoms of a heterocycle, but is not limited thereto.

[Chemical Formula 1]

.

.

More specifically, the "derivative of S-methylmethionine sulfonium" of the present invention is a derivative of (2S, 4S) and (2R, 4S) -2-phenylthiazolidine-4-carboxylic acid (2S, 4S), (2R, 4S) -2-phenylthiazolidine-4-carboxylic acid, or (2S, 4R) (2S, 4S) -2-phenyl-1,3-thiazin-4-carboxylic acid ((2S, 4R) 1,3-thiazinane-4-carboxylic acid.

(2)

; 및

; And

(3)

.

.

The " ultraviolet ray " of the present invention is preferably one of UVA, UVB, and UVC, and more preferably UVB, but is not limited thereto.

The " skin damage " of the present invention is preferably caused by UVB, and is preferably any one of an image, erythema or pigment deposition occurring in the epidermis or dermis, but is not limited thereto.

In a specific embodiment of the present invention, the present inventors selected compounds 15 and 16 from various compounds by selecting compounds capable of promoting the proliferation of skin cells and exhibiting protective effects against ultraviolet rays as SMMS derivatives (Fig. 1 And Fig. 2).

In addition, the present inventors confirmed that the compound 15 and the compound 16, which are the selected compounds, exerted on the skin fibroblast. As a result, both the compound 15 and the compound 16 promoted the proliferation of the dermal fibroblast (Figs. 3A and 3B) (Fig. 3C and Fig. 3D). The results are shown in Fig. In addition, Compound 15 and Compound 16 increase the level of mRNA expression of collagen synthase in UVB irradiated cells, inhibit the mRNA of melanin synthase (MMP-1), and cleave the intracellularly produced reactive oxygen species (Fig. 3E and Fig. 3F).

In addition, the present inventors confirmed that Compound 15 and Compound 16 can exhibit cytoprotective effect on ultraviolet rays, as well as a significant cell proliferation promoting effect on HaCaT cells, which are skin keratinocyte cell lines (FIGS. 4A to 4E).

(2S, 4S), (2S, 4S) -2-phenylthiazolidine-4-carboxylic acid (Compound 15) and (2S, 4R) (2R, 4R), (2R, 4S) -2-phenyl-1,3-thiazin-4-carboxylic acid (Compound 16) promotes cell proliferation in human dermal fibroblasts and dermal keratinocytes And exhibits a cytoprotective effect against ultraviolet rays. Therefore, it is effective as an effective ingredient of a cosmetic composition and a quasi-drug for skin damage prevention and skin protection by ultraviolet rays.

When the SMMS derivative compound of the present invention is used as an active ingredient of a cosmetic composition or a quasi-drug composition, the SMMS derivative compound exhibiting a skin-protecting effect by ultraviolet rays can be directly added or used together with other quasi-drugs or quasi-drugs, And the like. The amount of the active ingredient to be mixed can be appropriately determined depending on the purpose of use.

The cosmetic composition of the present invention may be in the form of a solution, a gel, a solid or a paste anhydrous product, an emulsion obtained by dispersing an oil phase in water, a suspension, a microemulsion, a microcapsule, a microgranule or an ionic (liposome) In the form of a foliar dispersant, in the form of creams, skins, lotions, powders, ointments, essences, sprays or conical sticks. It can also be prepared in the form of a foam or an aerosol composition further containing a compressed propellant.

The quasi-drug for skin damage prevention and skin protection by ultraviolet rays of the present invention is not particularly limited in its formulation and may be variously formulated in the form of a quasi-drug known in the art which shows skin protection effect by ultraviolet rays. The formulated product may be selected from the group consisting of sunscreen, sunscreen, sun lotion, sun spray, hair tonic, hair lotion, hair cream, hair spray, hair mousse, hair gel, hair conditioner, hair shampoo, hair rinse, Such as eyebrows, eyelashes, eyelashes, eyelashes, pet shampoo, pet rinse, hand cleaner, detergent soap, soap, disinfectant cleaner, wet tissue, mask, ointment, patch or filter filler, .

In addition, cosmetic compositions and quasi-drugs for skin damage prevention and skin protection by ultraviolet rays in each formulation may be blended with other ingredients arbitrarily selected depending on the formulation of other quasi-drugs, the purpose of use, and the like. The amount of the active ingredient to be mixed may be appropriately determined depending on the purpose of use, and may include, for example, customary adjuvants such as thickeners, stabilizers, solubilizers, vitamins, pigments and flavors, and carriers.

The content of the composition is preferably 0.0001 to 10% by weight based on the total weight of the composition. If it is more than 10% by weight, the color and stability of the composition are poor. When the composition is less than 0.0001% by weight, . The quasi-drug containing the SMMS derivative compound of the present invention as an active ingredient has little toxicity and side effects to cells as confirmed by the treatment index, and thus can be useful as a quasi-drug material.

The present invention also relates to a pharmaceutical composition for preventing or treating skin damage caused by ultraviolet rays, comprising a derivative of S-methylmethionine sulfonium (SMMS) or a pharmaceutically acceptable salt thereof as an active ingredient to provide.

The "S-methylmethionine sulfonium derivative" of the present invention is preferably a compound having a structure represented by the following formula (1), wherein R ₁ represents any one or both of S and N , 5 or 6 atoms of a heterocycle, but is not limited thereto.

[Chemical Formula 1]

.

.

More specifically, the "derivative of S-methylmethionine sulfonium" of the present invention is a derivative of (2S, 4S) and (2R, 4S) -2-phenylthiazolidine-4-carboxylic acid (2S, 4S), (2R, 4S) -2-phenylthiazolidine-4-carboxylic acid, or (2S, 4R) (2S, 4S) -2-phenyl-1,3-thiazin-4-carboxylic acid ((2S, 4R) 1,3-thiazinane-4-carboxylic acid.

(2)

; 및

; And

(3)

.

.

The " ultraviolet ray " of the present invention is preferably one of UVA, UVB, and UVC, and more preferably UVB, but is not limited thereto.

The " skin damage " of the present invention is preferably caused by UVB, and is preferably any one of an image, erythema or pigment deposition occurring in the epidermis or dermis, but is not limited thereto.

Compounds 15 and 16, derivatives of SMMS of the present invention, exhibit cell proliferation promoting effect in human dermal fibroblast and skin keratinocyte and exhibit cytoprotective effect against ultraviolet rays, so that prevention and treatment of skin damage by ultraviolet rays ≪ / RTI > is effective as an active ingredient of a pharmaceutical composition for < / RTI >

 The pharmaceutical composition for prevention and treatment of skin damage by ultraviolet rays of the present invention may contain one or more active ingredients which contain SMMS derivatives alone or exhibit similar functions to SMMS derivatives. The inclusion of additional ingredients may further enhance the skin protection effect of the composition of the present invention. When the above ingredients are added, skin safety, easiness of formulation, and stability of effective ingredients can be considered according to the combined use.

The pharmaceutical compositions of the present invention may include a pharmaceutically acceptable salt of the SMMS derivative. As used herein, the term " pharmaceutically acceptable " refers to those that are physiologically acceptable and do not normally cause an allergic reaction or similar reaction when administered to humans, wherein the salt includes a pharmaceutically acceptable free acid acid is preferred.

Acid addition salts include those derived from inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid or phosphorous acid, and aliphatic mono- and dicarboxylates, phenyl-substituted alkanoates, hydroxyalkanoates, Dioleate, aromatic acid, aliphatic and aromatic sulfonic acids. Such pharmaceutically innocuous salts include, but are not limited to, sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate chloride, bromide, Butyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, succinate, maleic anhydride, maleic anhydride, , Sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, Methoxybenzoate, phthalate, terephthalate, benzene sulfonate, toluene sulfonate, chlorobenzene sulfide Propyl sulphonate, naphthalene-1-yne, xylenesulfonate, phenylsulfate, phenylbutyrate, citrate, lactate,? -Hydroxybutyrate, glycolate, maleate, Sulfonate, naphthalene-2-sulfonate or mandelate.

The acid addition salt according to the present invention can be prepared by a conventional method, for example, by dissolving the SMMS derivative compound in an excess amount of an acid aqueous solution, and using this salt in a water-miscible organic solvent such as methanol, ethanol, acetone or acetonitrile Followed by precipitation. By heating the same amount of the SMMS derivative compound and the acid or alcohol in water, and then evaporating and drying the mixture, or by filtering the precipitated salt by suction filtration.

In addition, bases can be used to make pharmaceutically acceptable metal salts. The alkali metal or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess amount of an alkali metal hydroxide or an alkaline earth metal hydroxide solution, filtering the insoluble compound salt, and evaporating and drying the filtrate. At this time, it is preferable for the metal salt to produce sodium, potassium or calcium salt. The corresponding silver salt is also obtained by reacting an alkali metal or alkaline earth metal salt with a suitable salt (such as silver nitrate).

In addition, the SMMS derivative compound of the present invention includes not only pharmaceutically acceptable salts, but also all salts, hydrates and solvates which can be prepared by conventional methods.

The addition salt according to the present invention can be prepared by a conventional method, for example, by dissolving the SMMS derivative compound in a water-miscible organic solvent such as acetone, methanol, ethanol, acetonitrile or the like, adding an excess of organic acid, And then precipitating or crystallizing the acid solution. Subsequently, in this mixture, a solvent or an excess acid is evaporated and dried to obtain an additional salt, or the precipitated salt may be produced by suction filtration.

When the composition of the present invention is used as a medicament, the pharmaceutical composition containing the SMMS derivative compound or a pharmaceutically acceptable salt thereof as an active ingredient may be formulated into various oral or parenteral dosage forms at the time of clinical administration, But is not limited thereto.

Examples of the formulations for oral administration include tablets, pills, light / soft capsules, liquids, suspensions, emulsions, syrups, granules and elixirs. These formulations may contain, in addition to the active ingredient, a diluent (e.g., lactose, dextrose, (E.g., silica, talc, stearic acid and magnesium or calcium salts thereof and / or polyethylene glycols), such as, for example, water, rosin, sucrose, mannitol, sorbitol, cellulose and / or glycine. The tablets may also contain binders such as magnesium aluminum silicate, starch paste, gelatin, methylcellulose, sodium carboxymethylcellulose and / or polyvinylpyrrolidine and may optionally contain additives such as starch, agar, alginic acid or its sodium salt A disintegrating or boiling mixture and / or an absorbent, a colorant, a flavoring agent, and a sweetening agent.

Examples of transdermal dosage forms include ointments, creams, lotions, gels, solutions for external use, pastes, liniments, and air lozenges. In the above, 'transdermal administration' means that the pharmaceutical composition is locally administered to the skin, whereby an effective amount of the active ingredient contained in the pharmaceutical composition is delivered into the skin.

The pharmaceutical composition comprising the SMMS derivative compound of the present invention or a pharmaceutically acceptable salt thereof as an active ingredient can be administered parenterally, and parenteral administration can be carried out by injecting subcutaneous injection, intravenous injection, intramuscular injection, It depends on the method. In this case, in order to formulate the formulation for parenteral administration, the SMMS derivative compound or a pharmaceutically acceptable salt thereof is mixed with water or a stabilizer or a buffer to prepare a solution or suspension, which is prepared into an ampule or vial unit dosage form . The compositions may contain sterilized and / or preservatives, stabilizers, wettable or emulsifying accelerators, adjuvants such as salts and / or buffers for the control of osmotic pressure, and other therapeutically useful substances, Or may be formulated according to the coating method.

The dose of the compound of the present invention to the human body may vary depending on the patient's age, weight, sex, dosage form, health condition, and disease severity. In general, when referring to an adult patient weighing 60 kg, And may be 0.01 to 500 mg / day, preferably 0.01 to 500 mg / day, and may be administered once or several times a day at regular intervals according to the judgment of a doctor or pharmacist.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these examples are for illustrative purposes only and that the scope of the present invention is not construed as being limited by these examples.

hDF  Preparation and screening of novel compounds with cell proliferation promoting effect

<1-1> hDF  Selection of compound having proliferation effect

Novel compounds were synthesized for use as the subject compounds of the present invention to produce a total of 50 synthetic compounds.

Human dermal fibroblast (hDF) was then incubated with 1% fetal bovine serum (FBS; Gibco, Invitrogen, Carlsbad, CA, USA), 1% penicillin and streptomycin (Hyclone, Thermo Scientific, Logan, UT, USA), and the cells were cultured in a 5% carbon dioxide incubator at 37 ° C. For cell culture, the medium was replaced with fresh medium once every two days. The prepared hDF cells were inoculated into DMEM containing 10% FBS at a concentration of 3 × 10 ⁴ / well. After overnight incubation, starvation was induced by exchanging the culture medium with DMEM medium containing no serum. Each of the synthetic compounds was then treated with a concentration of 10 [mu] M and further incubated for 48 hours. During the cultivation, UVB (400 mJ) was treated or not treated and cultured separately. Then, 5 mg / ml MTT solution in PBS buffer was added at 5% volume of the medium added to each well, incubated at 37 캜 for 2 hours, and then the supernatant was removed. The precipitated insoluble formazan crystals of the wells from which the supernatant was removed were dissolved by adding dimethyl sulfoxide and the absorbance was measured at 595 nm using an ELISA reader to confirm the extent of the viable cells Respectively. Using the measured absorbance values, the degree of cell viability of the synthetic compound was calculated for a control without treatment of the synthetic compound. Using the calculated relative cell viability, the relative cell viability in the experimental group without UVB irradiation was confirmed as proliferation (%), and the relative cell viability in the UVB-irradiated experimental group was evaluated as cell viability ,%).

As a result, when the synthetic compound was treated at a concentration of 10 μM as shown in FIG. 1A and FIG. 1B, it was confirmed that the proliferation promoting effect was shown in the hDF cells significantly in the compounds 15, 16 and 22 (FIG. In addition, the survival rate of hDF was decreased when treated with UVB, but the survival rate of hDF was increased in the experimental group treated with compounds 5, 9, 15, 16, 17, 20 or 22 (FIG.

However, although Compound 5 and Compound 9 showed an effect of protecting hDF cells against UVB, it was confirmed that they had a disadvantage that they did not decompose and stink. Compounds 17, 20 and 22 were similar in structure to Compound 15 and Compound 16, but did not significantly exhibit cell proliferation promoting effect in an environment not irradiated with UVB, and as a more effective new compound, Compound 15 and Compound 16 Respectively.

<1-2> New  Synthesis and Characterization of Compound 15 and Compound 16

Among the various synthetic compounds, the structure of the compound was analyzed by NMR analysis for Compound 15 and Compound 16, which were found to be most effective in promoting proliferation of skin fibroblast cells and cytoprotective effect against UVB.

Specifically, Compound 15 and Compound 16 were synthesized while leading to the process of FIG. 0.74 mmol of L-cysteine or DL-homocysteine (DL-homocysteine) was mixed with 4 mL of ethanol and 1 mL of distillation, and then 0.74 mmol of benzaldehyde was added. The mixture was mixed at room temperature for 3 days and then filtered to remove the supernatant and a precipitate was obtained, washed with ether and dried under vacuum to give a white crystalline compound.

As a result, it was confirmed that the NMR analysis result of Compound 15 of the present invention was as follows: White solid; (D, J = 4.8, 4.8 Hz, 0.6H), 5.50 (s, 0.4H), 4.23 , 3.90 (dd, J = 7.6, 8.0Hz, 0.4H), 3.28-3.40 (m, 1H), 3.06-3.16 (m, 1H). That is, the compound 15 is (2S, 4S) and (2R, 4S) -2-phenylthiazolidine-4-carboxylic acid ((2S, 4S) -2-phenylthiazolidine-4-carboxylic acid):

(2)

.

In addition, it was confirmed that the NMR analysis results of Compound 16 of the present invention were as follows: White solid; 1H NMR (400 MHz, DMSO-d6)? 7.28-7.45 (m, 5H), 5.27 (s, IH), 3.60 (dd, J = 3.2,12 Hz, IH), 3.19-3.26 2.88-2.93 (m, 1 H), 2.04-2.08 (m, 1 H), 1.50 (m, 1 H). Accordingly, Compound 16 is a (2S, 4R), (2S, 4S), (2R, 4R) (2S, 4R) -2-phenyl-1,3-thiazinane-4-carboxylic acid (2S, 4R)

(3)

.

The effects of Compound 15 and Compound 16 on promoting dermal fibroblast proliferation and On UVB  Confirmation of cell protection effect

&Lt; 2-1 > Confirmation of promoting effect of the compound of the present invention on dermal fibroblast proliferation

In order to confirm the effect of the compound of the present invention in confirming the promoting effect of the proliferation of the dermal fibroblasts in an environment not irradiated with UVB, it was confirmed whether or not the dermal fibroblast proliferation promoting effect can be exhibited in a concentration dependent manner.

Specifically, MTT analysis was performed in the same manner as in Example <1-1> above and in a medium environment in which Compound 15 or 16 was treated at a concentration of 0.1, 1, 10, 50 or 100 μM, And whether or not the proliferation promoting effect of the cells was observed.

As a result, it was confirmed that Compound 15 and Compound 16 both stimulated the proliferation of dermal fibroblast cells in a concentration-dependent manner at a concentration of 0.1 to 100 μM as shown in FIGS. 3A and 3B (FIGS. 3A and 3B).

<2-2> UVB  Identification of the cytoprotective effect of the compounds of the present invention in an irradiation environment

In order to confirm the effect of the compound of the present invention in confirming the protective effect of dermal fibroblasts in an environment irradiated with UVB, it was confirmed whether or not the dermal fibroblast protection effect could be shown in a concentration-dependent manner.

Specifically, the same method as in the above Example <1-2> was conducted to irradiate 400 mJ of UVB, and the compound 15 or 16 was treated in a medium of 0.1, 1, 10, 50 or 100 μM MTT analysis was performed to determine whether the survival rate of dermal fibroblasts could be increased.

In addition, intracellular expression levels of collagen and MMP-1 enzyme were confirmed to confirm skin cell regeneration effect in cells. HDF cells treated with Compound 15 or Compound 16 were obtained, from which total RNA was extracted using a Trizol reagent. MRNA of collagen type I, MMP-1, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was synthesized by cDNA through reverse transcription PCR using the primers shown in Table 1 below. Then, 30 μl of the synthesized cDNA was used as a template, and PCR was performed again to amplify the corresponding DNA. PCR conditions were as follows: denaturation at 95 ° C for 5 min; 0 s at 95 째 C, 20 s at 54 째 C and 30 s at 72 째 C (35 total cycles); And final extension at 72 ° C for 10 minutes. GAPDH mRNA expression levels were used for strandarization of samples.

Untreated control (control) was a control group that was not treated with UVB, and the positive control (+) was a control group that did not treat the compound in the same environment irradiated with UVB.

The sequences of the primers used for confirmation of expression of collagen synthase and MMP-1 Name of the primer direction order SEQ ID NO: collagen_F Forward  5'-TAGG-GTCTAGACATGTTCAGCTTTGT-3 ' SEQ ID NO: 1 collagen_R Reverse  Gt; SEQ ID NO: 2 MMP-1_F Forward  5'-AGATGTGGAGTGCCTGATGT-3 ' SEQ ID NO: 3 MMP-1_R Reverse  5'-AG-CTAGGGTACATCAAAGCC-3 ' SEQ ID NO: 4 GAPDH_F Forward  5'-CGAGA-TCCCTCCAAAATCAA-3 ' SEQ ID NO: 5 GAPDH_R Reverse  5'-TGTGGTCATGAGTCCTCCCA-3 ' SEQ ID NO: 6

As a result, as shown in Figs. 3C, 3D and 3E, it was confirmed that Compound 15 and Compound 16 both showed an effect of protecting skin fibroblasts in a concentration-dependent UVB irradiation environment (Fig. 3C and Fig. 3D). In addition, when Compound 15 or Compound 16 was treated in an environment irradiated with UVB, the level of mRNA transcription of collagen enzyme was significantly increased as compared with the untreated control and positive control, and the MMP-1 enzyme synthesizing the melanin pigment Expression was significantly reduced in the Compound 15 or Compound 16 treated group compared to the positive control and restored to express MMP-1 at a similar level as the untreated control, so that Compound 15 and Compound 16 inhibited cell damage by UVB irradiation (Fig. 3E). &Lt; RTI ID = 0.0 &gt;

< 2-3> UVB  In the irradiation environment, Compound 15 and Compound 16 Generated Active oxygen species (ROS) reduction effect

Compounds 15 and 16 of the present invention were found to exhibit significant cytoprotective effects in the UVB irradiation environment and it was confirmed whether or not ROS generated in the UVB environment can be reduced.

Specifically, ROS generation was measured using a DCFnDA kit (Molecular Probes, Eugene, OR). The αMEM medium containing 0.2% FBS was dispensed into a 6-well plate, and hDF cells were inoculated thereto. Inoculated cells were treated with 10 [mu] M Compound 15 or Compound 16, and 20 [mu] M DCFDDA in the UVB irradiation environment and cultured. Culturing was carried out under the usual culture conditions, and the fluorescence of DCF-DA was measured by IncuCyte (Essen Bioscience, MI) every 10 minutes, and each well was imaged. The fluorescence intensity of the measured DCF-DA was quantitatively calculated using IncuCyteTM software.

As a result, as shown in FIG. 3F, the generation of ROS was induced by UVB in hDF, but when the compound 15 or compound 16 was treated, the level of ROS in the cells was significantly decreased, It was confirmed that Compound 15 and Compound 16 can exhibit scavenging activity against ROS produced in hDF (Fig. 3F).

The dermal keratinocytes of compound 15 and compound 16 keratinocyte ) Proliferation promoting effect and On UVB  Confirmation of cell protection effect

It was confirmed that Compound 15 and Compound 16 of the present invention can exhibit significant proliferation promoting effect and cytoprotective effect against UVB damage on dermal fibroblast cell line, And HaCaT cells were confirmed to have the same effect as above.

Specifically, human-derived HaCaT cells, which are dermal keratinocyte cell lines, were cultured in a DMEM medium containing 10% FBS, and then cultured in KGM2 (Keratinosute Basal Medium 2), and cultured in the same manner as in Example <1-3> Compound 15 or Compound 16 was treated and MTT assay was performed to confirm cell proliferation promoting effect and cytoprotective effect against UVB. In addition, ROS scavenging activity of Compound 15 and Compound 16 against intracellular ROS produced by UVB irradiation was confirmed by the same method as Example <2-3.

As a result, Compound 15 did not significantly increase HaCaT cell proliferation as shown in FIGS. 4A to 4D (FIG. 4A). However, for cell damage by UVB irradiation of 500 mJ, (Fig. 4C). On the contrary, it was confirmed that Compound 16 not only promoted the proliferation of HaCaT cells in a concentration-dependent manner (FIG. 4B) but also showed a significant cytoprotective effect in the UVB irradiation environment (FIG. 4D). In addition, it was confirmed that both of Compound 15 and Compound 16 can erase ROS generated in HaCaT cells significantly in the ROS scavenging activity generated in the cells by UVB (Fig. 4E).

<110> Industry-Academic Cooperation Foundation, Yonsei University <120> Cosmetic composition for protecting skin and inhibiting damage          skin against ultraviolet comprising S-Methylmethionine sulfonium          derivative <130> 1061033 <160> 6 <170> Kopatentin 2.0 <210> 1 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> collagen_R <400> 1 tagggtctag acatgttcag ctttgt 26 <210> 2 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> collagen_R <400> 2 gtgattggtg ggatgtcttc gt 22 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> MMP-1_F <400> 3 agatgtggag tgcctgatgt 20 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> MMP-1_R <400> 4 agctagggta catcaaagcc 20 <210> 5 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> GAPDH_F <400> 5 cgagatccct ccaaaatcaa 20 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> GAPDH_R <400> 6 tgtggtcatg agtcctccca 20

Claims (10)

delete delete (2S, 4R), (2S, 4S), (2R, 4R) or (2R, 4S) -2-phenyl-1,3-thiazin- (2R, 4S) -2-phenyl-1,3-thiazinane-4-carboxylic acid, or a pharmaceutically acceptable salt thereof, is effective And is characterized in that no odor is generated. The cosmetic composition for skin damage prevention and skin protection by ultraviolet rays:
(3)

.
The cosmetic composition according to claim 3, wherein the ultraviolet ray is UVB.
4. The cosmetic composition according to claim 3, wherein the skin damage caused by the ultraviolet rays is any one of an image, erythema or pigment deposition generated in the epidermis or dermis by ultraviolet rays.
delete delete (2S, 4R), (2S, 4S), (2R, 4R) or (2R, 4S) -2-phenyl-1,3-thiazin-4-carboxylic acid represented by the following formula A pharmaceutical composition for preventing or treating skin damage caused by ultraviolet rays, which comprises a pharmaceutically acceptable salt thereof as an active ingredient and does not generate an offensive odor.
(3)

.
The pharmaceutical composition according to claim 8, wherein the ultraviolet light is UVB.
The pharmaceutical composition for preventing or treating skin damage caused by ultraviolet rays according to claim 8, wherein the skin damage caused by ultraviolet rays is any one of an image, erythema or pigment deposition generated on the epidermis or dermis by ultraviolet rays .

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