KR20220168292A - Skin whitening pharmacological composition with pickpeltaraenin - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for skin whitening using Picfeltarraenin IA as an active component. The Picfeltarraenin IA increases the total amount of collagen in skin fibroblasts to promote skin regeneration, and inhibits the production of nitric oxide (NO), which is involved in the inflammatory response to have an excellent anti-inflammatory effect and a whitening effect that significantly reduces the total amount of melanin, thereby being able to be used in pharmaceuticals or cosmetics.

Description

Skin whitening pharmaceutical composition using pickpeltaraenin IA as an active ingredient {Skin whitening pharmacological composition with pickpeltaraenin}

The present invention relates to a composition for skin regeneration, wrinkle improvement, anti-inflammatory or skin whitening.

Collagen is a major matrix protein produced by fibroblasts of the skin and exists in the extracellular matrix.

Mechanical firmness of the skin, resistance and cohesiveness of connective tissue, support of cell adhesion, cell division and differentiation (growth of organisms)

or wound healing), etc. are known. Such collagen is reduced by age and photoaging by UV irradiation.

It is known that it is closely related to the formation of wrinkles in the skin. In addition, in recent years, the light on skin aging

Advances in extensive research are revealing important functions of collagen in the skin.

Active ingredients exhibiting an anti-wrinkle effect by promoting collagen synthesis are known. For example, retinoic acid

acid), TGF (transforming growth factor) [Non-Patent Document 1], protein derived from animal placenta [Patent Document 1], betulinic acid

(betulinic acid) [Patent Document 2] and chlorella extract [Patent Documents 3 and 4] are known as substances promoting collagen synthesis.

all. However, the active ingredients are limited in usage due to safety issues such as irritation and redness when applied to the skin, or

is insignificant, so the effect of improving skin function by promoting collagen synthesis in the skin cannot be expected.

there is.

In addition, inflammation is the body's immune response to injury or disease, and oxidative stress such as ultraviolet rays, active oxygen, and free radicals

Stress activates inflammatory factors and causes various diseases and aging of the skin. Kinin, a vasoactive polypeptide

(kinin), plasmin, or complement, etc., act on vasodilation, contraction, and chemotaxis.

In addition, lymphokines such as interleukin-6 (IL-6) and arachidonic acid induce inflammatory responses.

take charge Arachidonic acid is produced by two enzymes, cyclooxygenase and lipooxygenase.

Through this pathway, it is metabolized into prostaglandin or leukotrienes, which are inflammatory mediators, and various

mediates the inflammatory response.

On the other hand, anti-inflammatory agents are those that act to remove inflammatory sources and reduce biological responses and symptoms in order to disappear inflammation.

all. Substances that have been used for anti-inflammatory purposes so far include non-steroidal flufenamic acid,

There are profen (ibuprofen), benzydamine (benzydamine) or indomethacin (indomethacin), etc.

These include prednisolone or dexamethasone. Also allantoin, azene or hydro

Although cortisone and the like are known to be effective in anti-inflammatory, these substances are not used due to safety issues on the skin.

There are limitations or the effect is insignificant, so there is a problem in that the anti-inflammatory effect cannot be expected.

In addition, it is a common desire to have white and fair skin. The color or lightness of skin is the amount of melanin in a person's skin.

It is genetically determined according to the concentration and distribution of melanin, but environmental or biological factors such as solar ultraviolet rays, fatigue or stress

It is also affected by physical conditions. Melanin is derived from tyrosine, a type of amino acid.

After the enzyme e) acts as a catalyst and sequentially converts into DOPA and dopaquinone, a non-enzymatic acid

It is created through a chemical reaction. As such, the pathway by which melanin is produced is known, but the mechanism that induces melanin synthesis is known.

It is still not clear what causes tyrosinase to be triggered in the mechanism.

On the other hand, as a commonly known whitening ingredient, tyrosinase enzymes such as Kojic acid or Arbutin

Substances that inhibit deactivation, hydroquinone, vitamin C (L-Ascorbic acid) or their derivatives, and various

There are plant extracts. By inhibiting the synthesis of melanin pigment, they can realize skin whitening by brightening the skin tone.

In addition, it is possible to improve skin hyperpigmentation such as spots and freckles caused by ultraviolet rays, hormones, or heredity.

Do. However, when applied to the skin, there is a limit to the amount used due to safety issues such as irritation and redness, or the effect is insignificant, so the actual

There is a problem in which no positive effect can be expected.

Therefore, it is safe for the living body, active ingredients are stable, and above all, existing skin regeneration, wrinkle improvement, anti-inflammatory or whitening effect

There is an urgent need to develop ingredients with skin regeneration, wrinkle improvement, anti-inflammatory or whitening activities that are more effective than substances with excess effects.

It is being desired.

Accordingly, the present inventors have found that Picfeltarraenin IA increases the total amount of collagen in fibroblasts of the skin,

It promotes the regeneration of parts, improves wrinkles, inhibits the production of nitric oxide (NO) to show anti-inflammatory effects, and the total amount of melanin

It was confirmed that the whitening effect was significantly reduced and the present invention was completed.

Therefore, an object of the present invention is skin regeneration, anti-wrinkle, anti-aging,

To provide a composition for inflammation or skin whitening:

[Formula 1]

As a means for solving the above problems, the present invention

A drug for skin regeneration, wrinkle improvement, anti-inflammatory or skin whitening comprising a compound represented by Formula 1 below as an active ingredient

A scientific composition is provided.

[Formula 1]

As another means for solving the above problems, the present invention

Skin regeneration, wrinkle improvement, anti-inflammatory or skin whitening formulation comprising a compound represented by Formula 1 as an active ingredient

A dressing composition is provided.

[Formula 1]

Picfeltarraenin IA according to the present invention increases the total amount of collagen in fibroblasts of the skin,

It promotes regeneration, suppresses the production of nitric oxide (NO) involved in inflammatory reactions, and has excellent anti-inflammatory effects, and the total amount of melanin

It can be used in pharmaceuticals or cosmetics by showing a whitening effect that significantly reduces

Hereinafter, the configuration of the present invention will be described in detail.

In order for skin regeneration, wrinkle improvement, anti-inflammatory or whitening ingredients to be effective when applied to the actual skin, low concentrations are required.

It exhibits highly active skin regeneration, wrinkle improvement, anti-inflammatory or whitening activity, and its ability to penetrate and absorb through the skin is dominant.

It is desirable to be able to In addition, it stays on for a sufficient time to exhibit skin regeneration, wrinkle improvement, anti-inflammatory or whitening effects.

It has low volatility so that it can be softened, and the active ingredient is stably maintained on the composition or skin, and it can be used in medicine or cosmetics.

It is easy to formulate into the furnace, and it is also desirable that it is safe for the skin. However, among known components, all of the above properties are satisfied.

Ingredients are rare. For example, some skin regenerating, wrinkle-reducing, anti-inflammatory or whitening ingredients have been tested in vitro.

Even at low concentrations, skin regeneration, wrinkle improvement, anti-inflammatory or whitening activities are excellent, but the ability to penetrate and absorb through the skin is poor.

It is difficult to apply to actual skin. Other active ingredients have low hydrophilicity, making them difficult to formulate as medicines or cosmetics.

difficult. In addition, some skin regenerating, wrinkle-reducing, anti-inflammatory or whitening ingredients are damaged when exposed to heat, light or oxygen.

In some cases, the active ingredient is decomposed or transformed into other compounds, and the effect is already lost before application to the skin.

As can be seen in the following examples, Picfeltarraenin IA has a total amount of collagen at low concentrations.

It shows the effect of significantly increasing the amount of melanin, the effect of significantly reducing the total amount of melanin, and the effect of inhibiting the production of nitric oxide.

It can be used as an active ingredient in pharmaceuticals, cosmetics, or health foods for regeneration, wrinkle improvement, anti-inflammatory, or skin whitening.

there is.

Therefore, the present invention is a skin regeneration, wrinkle improvement, anti-inflammatory or

Provides a pharmaceutical composition for skin whitening.

[Formula 1]

The compound of Formula 1 is called Picfeltarraenin IA, and may include all possible isomers.

can

The compound of Formula 1 may be used after being isolated or synthesized from plant extracts, or a commercially available compound may be used.

can

The pharmaceutical composition for skin regeneration, wrinkle improvement, anti-inflammatory or skin whitening of the present invention is a pharmaceutical composition of the compound of Formula 1.

It may contain acceptable salts.

A pharmaceutically acceptable salt of the compound of Formula 1 is an acid addition salt formed using an organic acid or an inorganic acid.

The organic acid may be, for example, formic acid, acetic acid, propionic acid, lactic acid, butyric acid, isobutyric acid, triple

Luoroacetic acid, malic acid, maleic acid, malonic acid, fumaric acid, succinic acid, succinic acid monoamide, glutamic acid, tartaric acid, oxalic acid,

Citric acid, glycolic acid, glucuronic acid, ascorbic acid, benzoic acid, phthalic acid, salicylic acid, anthranilic acid, dichloroacetic acid,

and salts based on minooxy acetic acid, benzenesulfonic acid, p-toluenesulfonic acid or methanesulfonic acid. Inorganic acids are, for example, hydrochloric acid,

hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, carbonic acid or boric acid based salts. preferably in the form of a hydrochloride or acetate salt

And, more preferably, it may be in the form of a hydrochloride salt.

The above-mentioned acid addition salt is obtained by a) directly mixing the compound of formula 1 and an acid, or b) using one of them as a solvent or

is dissolved in a water-containing solvent and mixed, or c) the compound of Formula 1 is placed in a solvent or an acid in a submerged solvent.

It is prepared by a general salt production method of mixing them.

In addition to the above, additionally saltable forms include gaba salt, gabapentin salt, pregabalin salt, nicotinate, adipate salt,

Hemimalonate, cysteine salt, acetylcysteine salt, methionine salt, arginine salt, lysine salt, ornithine salt or aspartate

acid salts, etc.

In addition, when the compound represented by Formula 1 of the present invention is used as a drug, it additionally exhibits the same or similar function.

may contain one or more active ingredients. For example, known skin regeneration, wrinkle improvement, anti-inflammatory or skin whitening ingredients

will be able to include When additional skin regeneration, wrinkle improvement, anti-inflammatory or skin whitening ingredients are included, the present invention

The skin regeneration, wrinkle improvement, anti-inflammatory or skin whitening effects of the composition of may be further enhanced. Add above ingredients

In case of complex use, skin safety, ease of formulation, and stability of active ingredients can be considered. one of the present invention

In embodiments, the composition is a skin regeneration component known in the art, retinoic acid, TGF, protein derived from animal placenta,

betulinic acid or chlorella extract; As a whitening ingredient known in the art, Kojic acid or Arbutin

Substances that inhibit tyrosinase enzyme activity, such as hydroquinone, vitamin-C (L-Ascorbic acid); this

derivatives of; and one or two or more components selected from the group consisting of various plant extracts.

can Additional components may be included in an amount of 0.0001% by weight or more and 10% by weight or less based on the total weight of the composition.

For example, 0.0001 wt% or more and 1 wt% or less, 0.0001 wt% or more and 0.1 wt% or less, 0.0001 wt% or more

to 0.001% by weight or less, 0.001% by weight or more to 10% by weight or less, 0.001% by weight or more to 1% by weight or less, 0.001% by weight

It may be greater than or equal to 0.1% by weight, greater than or equal to 0.01% by weight and less than or equal to 10% by weight, or greater than or equal to 0.01% by weight and less than or equal to 1% by weight.

The content range may be adjusted according to requirements such as skin safety and ease of formulation of the compound of Formula 1.

will be

In addition, the pharmaceutical composition for skin regeneration, wrinkle improvement, anti-inflammatory or skin whitening of the present invention may further include a pharmaceutically acceptable carrier.

Pharmaceutically acceptable carriers include buffer, sterile water for injection, normal saline or phosphate buffered saline, sucrose, histi

It may contain several components such as din, or polysorbate.

The pharmaceutical composition of the present invention can be administered orally or parenterally, in the form of a general pharmaceutical preparation, for example, clinical

When administered, it can be administered in various oral or parenteral formulations. In the case of formulation, commonly used fillers,

It may be formulated using diluents or excipients such as extenders, binders, wetting agents, disintegrants or surfactants.

Solid dosage forms for oral administration include tablets, pills, powders, granules or capsules, etc.

At least one excipient in the pharmaceutical composition of the name, for example, starch, calcium carbonate (Calcium carbonate), suc

It may be prepared by mixing sucrose, lactose, or gelatin.

In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. As a liquid formulation for oral use,

Suspensions, solutions for internal use, emulsions, or syrups are applicable. Water or liquid paraffin, which are commonly used simple diluents, are applicable.

In addition, various excipients, for example, wetting agents, sweeteners, fragrances, or preservatives may be included.

Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized formulations or suppositories.

Non-aqueous solvents or suspending solvents such as propylene glycol, polyethylene glycol or olive oil

silver vegetable oil; Injectable esters such as ethyl oleate and the like may be used. The base of the suppository is Witepsol

(witepsol), macrogol, tween 61, cacao fat, laurin fat or glycerogeratin, and the like can be used.

In the present invention, the term 'skin regeneration effect' refers to the recovery of skin tissue against damage caused by external and internal skin causes.

says that Damage caused by external causes may include ultraviolet rays, external contaminants, wounds or trauma,

Damage caused by the internal cause may include stress or the like.

In the present invention, the term 'wrinkle improvement effect' means suppressing or inhibiting the formation of wrinkles on the skin, or

It means to alleviate wrinkles.

In the present invention, 'anti-inflammatory effect' refers to suppressing inflammation. The inflammation is a biological response to any stimulus

As one of the protective responses of the body, it refers to a complex lesion in which three things are combined: tissue deterioration, circulatory disorder, exudation, and tissue proliferation.

all. More specifically, inflammation is part of innate immunity, and in humans, as in other animals, innate immunity is pathogen-specific.

Recognize patterns on the cell surface that exist as Phagocytes recognize cells with such surfaces as non-self and kill pathogens.

attack If a pathogen breaks through the body's physical barrier, an inflammatory response occurs. Inflammatory response at the wound site

It is a non-specific defense action that creates a hostile environment for invading microorganisms. In inflammatory reactions, wounds or external infections

When a body enters the body, leukocytes responsible for the early stage immune response flock to express cytokines. thus

The expression level of intracellular cytokines is an indicator of the activation of the inflammatory response. An example of a skin disease related to inflammation is atopic skin

erythematous diseases caused by inflammation, psoriasis, radiation, chemicals or burns; Acid burns, bullous dermatosis, lichenoid type diseases

or itching due to allergy or the like; Seborrheic eczema, rose acne, pemphigus vulgaris, exudative erythema multiforme, erythema nodosum,

inflammatory hair loss such as balanitis, vulvitis or alopecia areata; Including but limited to cutaneous T-cell lymphoma

is not

In the present invention, 'whitening effect' means not only brightens the skin tone by inhibiting the synthesis of melanin pigment, but also

It refers to improving skin hyperpigmentation such as spots or freckles caused by external rays, hormones, or heredity.

The pharmaceutical composition of the present invention, when containing an effective amount of the compound of Formula 1, is suitable for skin regeneration, wrinkle improvement, and anti-aging.

It may provide effects such as inflammation or whitening. In the present invention, the term 'effective amount' promotes the regeneration of damaged skin.

It refers to the amount of a compound capable of improving wrinkles, inhibiting inflammation, or exhibiting a whitening effect. the present invention

The effective amount of the compound of Formula 1 included in the composition of

It will depend on the method used or how long it stays on the skin. For example, the composition is commercialized as a pharmaceutical

In the case of formula 1 at a higher concentration than when commercialized as a cosmetic that is routinely applied to the skin.

may include a compound of Therefore, the daily dosage is 0.1 to 0.1 based on the amount of the compound of Formula 1 above.

100 mg/kg, preferably 30 to 80 mg/kg, more preferably 50 to 60 mg/kg, 1 to 6 times a day

can be administered.

The pharmaceutical composition of the present invention may be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy or biological response modifiers.

The present invention also relates to skin regeneration, wrinkle improvement, anti-inflammatory or whitening skin comprising the compound of Formula 1 as an active ingredient.

It can be provided in the form of a supplementary external preparation.

When the compound of Formula 1 is used as an external skin preparation, a fatty substance, an organic solvent, a solubilizing agent, a thickening agent and a gelling agent are additionally used.

agent, softener, antioxidant, suspending agent, stabilizer, foaming agent, fragrance, surfactant, water, ionic emulsifier,

Non-ionic emulsifier, filler, sequestering agent, chelating agent, preservative, vitamin, blocker, humectant, essential oil,

Any other dyes, pigments, hydrophilic actives, lipophilic actives or lipid vesicles commonly used in external skin preparations.

It may contain adjuvants commonly used in the field of dermatology, such as ingredients. In addition, the above ingredients are dermatological science

It can be introduced in an amount generally used in the field.

When the compound of Formula 1 is provided as an external skin preparation, it is not limited thereto, but ointments, patches, gels, creams or

It may be a formulation such as a spray.

The present invention also relates to a skin regeneration, wrinkle improvement, anti-inflammatory or whitening agent comprising the compound of Formula 1 as an active ingredient.

It can be provided in the form of a dressing.

When the compound of Formula 1 is used as a cosmetic, it is prepared by containing the compound of Formula 1 as an active ingredient.

Cosmetics can be prepared in the form of a general emulsified formulation or solubilized formulation. For example, softening lotion or nutrition

cosmetic water such as cosmetic water; emulsions such as facial lotions or body lotions; Nourishing Cream, Moisturizing Cream or Eye Cream

creams such as; essence; cosmetic ointment; spray; gel; pack; sunscreen; makeup base; liquid type, solid type

is a foundation such as a spray type; powder; makeup removers such as cleansing creams, cleansing lotions, and cleansing oils;

It may have formulations such as detergents such as cleansing foam, soap, body wash, and the like.

In addition to the compound of Formula 1, the cosmetics include a fatty substance, an organic solvent, a solubilizing agent, a thickening agent, a gelling agent, a softening agent,

Antioxidant, suspending agent, stabilizer, foaming agent, fragrance, surfactant, water, ionic emulsifier, nonionic oil

topical agent, filler, sequestering agent, chelating agent, preservative, vitamin, blocker, humectant, essential oil, dye, pigment, parent

It may contain adjuvants commonly used in the field of cosmetics, such as aqueous actives, lipophilic actives or lipid vesicles.

there is.

When the compound of Formula 1 is commercialized as a cosmetic, the active ingredient stays on the skin for a short period of time.

In the case of wash-off type cosmetics such as up removal agents or detergents, the above chemical formula is relatively high in concentration.

1 may be included. On the other hand, lotion, lotion, cream or

In the case of leave-on type cosmetics such as essence, a lower concentration of the above than wash-off type cosmetics

It may also include the compound of Formula 1. Although not limited thereto, in one embodiment of the present invention, the

The composition is 0.0001% to 10% by weight (preferably 0.0001% by weight) of the compound of Formula 1 based on the total weight of the composition.

weight% to 1% by weight). The composition of the present invention contains less than 0.0001% by weight of the compound of Formula 1.

When included, sufficient skin regeneration, wrinkle improvement, anti-inflammatory or whitening effects cannot be expected, and it exceeds 10% by weight

If it is included, unwanted reactions such as allergies may occur or there may be problems with skin safety.

It is for

The present invention also relates to a health food for skin regeneration, wrinkle improvement, anti-inflammatory or whitening containing the compound of Formula 1.

will be.

In the present specification, 'health food' refers to the compound of Formula 1 in food materials such as beverages, teas, spices, chewing gum, and confectionery.

Food additives, capsules, powders, suspensions, etc., that have certain health effects when ingested.

However, unlike general drugs, it is made from food and does not have side effects that can occur when taking drugs for a long time.

There are advantages.

Since the health food of the present invention obtained in this way can be consumed on a daily basis, high skin regeneration,

Wrinkle improvement, anti-inflammatory or whitening effects can be expected, so it is very useful.

When the compound of Formula 1 is used as a food additive, the compound of Formula 1 is added as it is or other formulas are used.

It can be used together with products or food ingredients, and can be used appropriately according to conventional methods. Mixed amount of active ingredient

may be appropriately determined according to its purpose of use (prevention, health or therapeutic treatment). In general, when preparing food or beverage, the composition of the present invention is added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less, based on the raw material.

do. However, when ingested for a long period of time for health and hygiene purposes, the content below the above range may be included.

Since there is no problem in malleability, the active ingredient may be used in an amount greater than the above range.

There is no particular limitation on the type of food. Examples of foods to which the above substances can be added include meat, sausage, bread, and chocolate.

Lit, candy, snack, confectionery, pizza, ramen, other noodles, chewing gum, dairy products including ice cream, various soups, beverages

There are water, tea, drinks, alcoholic beverages or vitamin complexes, and all health foods in the usual sense

include

The health beverage composition of the present invention contains various flavoring agents or natural carbohydrates as additional components, like conventional beverages.

can be useful The aforementioned natural carbohydrates include monosaccharides such as glucose and fructose; Disaccharides such as maltose and sucrose

ride; polysaccharides such as dextrins and cyclodextrins; Sugar alcohols such as xylitol, sorbitol, and erythritol

can Sweeteners include natural sweeteners such as thaumatin and stevia extract; Synthetic sweeteners such as saccharin and aspartame

can be used. The proportion of the natural carbohydrate is generally about 0.01 to 0.04 g per 100 mL of the composition of the present invention,

Preferably it is about 0.02-0.03 g.

In addition to the above, the health food of the present invention contains various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid, pectic acid salts, eggs

Gic acid, salts of alginic acid, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols or charcoal

It may contain an oxidizing agent and the like. In addition, the health food of the present invention is a natural fruit juice, fruit juice beverage, or vegetable beverage

May contain fruit flesh for manufacturing. These components may be used independently or in combination. these additives

Although the ratio is not very important, it is selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

Common.

Hereinafter, the present invention will be described in detail by examples. However, the following examples are only illustrative of the present invention,

The content is not limited to the following examples.

Reference Example 1: Picfeltarraenin IA (Picfeltarraenin IA) material information]

Designation: Picfeltarraenin IA

CAS No.

: 97230-47-2

Where to buy: Tauto Biotech Co., Ltd.

Example 1: Effect of increasing the total amount of collagen

The materials in Table 1 below were added to the culture medium of human-derived fibroblasts to measure the total amount of collagen at the cellular level. Cola

The total amount of gen was quantified using PICP EIA kit (Procollagen Type I C-Peptide Enzyme ImmunoAssay KIT).

all. Concentrations of test substances 10 ppm, 1 ppm, 0.1 ppm, 0.01 ppm, 0.001 ppm for human-derived fibroblasts before the experiment

Evaluation of cytotoxicity in (method of MTT test by culturing fibroblasts [Reference: Mossman T. (1983). Rapid

Colorimetric Assay for Cellular Growth & Survival: application to proliferation & cytotoxicity assays.

Journal of Immunological Methods 65, 55-63], and the total amount of collagen was measured by selecting a concentration without cytotoxicity.

In the experiment, the final concentrations of the compound of Formula 1 were 1 ppm and 10 ppm, respectively, and each sample was human fibroblast

After being added to the culture medium and cultured for 1 day, the culture medium was taken and the total amount of collagen at each concentration was analyzed with the PICP EIA kit.

It was measured at 450 nm using a photophotometer. Culture of fibroblasts without adding anything for comparison of effects

The total amount of collagen was determined in the same way for the sample in which the medium (control group) and vitamin C were added to a final concentration of 52.8 ppm.

measured. The total amount of collagen was measured as UV absorbance, and the increase rate of the total amount of collagen was calculated as the relative collagen to the control group.

Calculated by the ratio of the total amount of gen, and the results are summarized in Table 1 below.

Table 1

The effect of increasing the total amount of collagen of the compound of Formula 1 (number of repetitions = 3)

Sample absorbance average collagen increase rate (%)

Compound of Formula 1

(1ppm)

551 123.0

Compound of Formula 1

(10 ppm)

770 171.8

control group

(no additives)

448 -

vitamin C

(52.8 ppm)

581 129.7

As can be seen from the results of Table 1, the compound of Formula 1 contains vitamin C, which is known to have the ability to synthesize collagen.

A better increase in the total amount of collagen was shown at a lower concentration than when applied.

Therefore, it was found that the compound of Formula 1 can be used for skin regeneration and wrinkle improvement.

Example 2: Collagenase activity inhibitory effect

For the substances in Table 2 below, the effect of inhibiting collagenase activity was confirmed as follows.

Prior to the experiment, human-derived fibroblasts were tested at concentrations of 10 ppm, 1 ppm, 0.1 ppm, 0.01 ppm, and 0.001 ppm.

Evaluation of cytotoxicity (method for MTT test by culturing fibroblasts [Reference: Mossman T. (1983). Rapid

Colorimetric Assay for Cellular Growth & Survival: application to proliferation & cytotoxicity assays.

Journal of Immunological Methods 65, 55-63], and a method for evaluating collagenase by selecting a concentration without cytotoxicity

was performed.

Fibroblasts, which are human normal skin cells, were cultured in a 24-well microplate at 2.5 × 10 per well.

4

Inoculate to become cells,

After culturing for 24 hours in 10% serum DMEM medium and 37 ° C, the 10% serum DMEM medium was removed and phosphate buffered

After washing once with the serum-free DMEM medium to which the compound of formula 1 was added and the compound of formula 1 was not included as a control.

They were further incubated for 30 minutes in serum-free DMEM medium.

TNF-α (tumor necrosis factor-α), a substance known to produce MMP-1, a collagenase, after 30 minutes of sample treatment

After stimulation with 50 ng/ml, it was cultured for 24 hours. At this time, the TNF-α untreated group and the treated group did not contain the compound of Formula 1.

Among the control group, a non-treated group not treated with TNF-α and a treated group treated with TNF-α were used.

The supernatant of each well was collected and the amount (ng/ml) of newly synthesized MMP-1 was measured using an MMP-1 assay kit (Amersham, USA).

High, collagenase activity inhibition rate was calculated as MMP-1 production inhibition rate (%) according to Equation 1 below, the results are shown in the table below

As shown in 2.

The amount of MMP-1 in the experimental group refers to the group to which the substance was added at each concentration.

Table 2

Collagenase activity inhibitory effect of the compound of Formula 1 (number of repetitions = 4)

Sample Amount of MMP-1 (ng/ml) Inhibition rate (%)

Compound of Formula 1 10 ppm 6.89 93.03%

Compound of Formula 1 1 ppm 17.02 13.17%

Negative control group (TNF-α untreated) 5.99 -

Positive control group (TNF-α treatment) 18.90 -

As can be seen from the results of Table 2, when the compound of formula 1 and TNF-α were treated, the compound of formula 1 was treated

It can be seen that the production of MMP-1 was decreased compared to the positive control group treated with TNF-α without treatment. That is, the compound of Formula 1

exhibited an excellent collagenase activity inhibitory effect.

Therefore, it was found that the compound of Formula 1 can be used for skin regeneration and wrinkle improvement.

Example 3: Whitening effect - melanin total amount reduction effect

By adding the compound of formula 1 to the culture medium of mouse melanoma cells (B-16 mouse melanoma cells), melanoma at the cellular level

The whitening effect was confirmed by measuring the total amount of nin (Lotan R., Lotan D. Cancer Res. 40:3345-3350, 1980). Experiment

Toxicity was evaluated for melanoma cells of all mice, and a non-toxic concentration was selected to perform whitening evaluation.

The compound of Formula 1 was added to the culture medium to a final concentration of 5 ppm, 10 ppm, and 20 ppm, and arbutin, a control, was 200

It was added to the medium so as to be ppm, treated with B-16 melanoma cells, respectively, and cultured for 3 days.

Thereafter, the cells were treated with trypsin, detached from the culture vessel, centrifuged, and then melanin was extracted. removed

Cells were boiled for 10 minutes by adding 1 mL of sodium hydroxide solution (1N concentration) to dissolve melanin, and using a spectrophotometer, 400

The amount of melanin produced was measured by measuring the absorbance at nm.

The amount of melanin per unit cell number (1 × 10

6

It was measured by the method of expressing the absorbance of cell), and the phase relative to the control

The total amount of melanin was calculated as an inhibition rate (%), and the results are summarized in Table 3. The experiment was repeated 3 times.

Table 3

The effect of reducing the total amount of melanin at the cellular level according to the concentration

Total amount of sample melanin (Abs.) Inhibition rate (%)

Control (no additives) 0.189 -

Control 1: Arbutin (200 ppm) 0.120 36.5

Compound of Formula 1 (20 ppm) 0.088 53.4

Compound of Formula 1 (10 ppm) 0.101 46.6

Compound of Formula 1 (5 ppm) 0.150 20.6

[0092] As can be seen from the results of Table 3, the compound of Formula 1 exhibits a lower total amount of melanin for mouse melanoma cells cultured at a lower concentration than arbutin, known as a whitening substance, and is thus used for whitening purposes. know you can use

could

Example 4: Anti-inflammatory effect-NO production inhibitory effect

In order to confirm the anti-inflammatory effect and skin trouble relieving effect of the compound of Formula 1, the RAW264.7 cell line (ATCC number:

CRL-2278) was used to test nitric oxide (NO) formation inhibition by the GRIESS method.

Specifically, mouse macrophage RAW264.7 cells were subcultured several times, and 3 × 10 cells were cultured in one well.

5

24-

After being put in the well plate, it was cultured for 24 hours. Then, the compound of Formula 1 was diluted at the concentration shown in Table 4 below.

was analyzed and replaced with the cell medium containing At this time, L-NMMA (L-NG-Monomethylarginine), an NO-production inhibitor, was positive.

It was treated together as a control and incubated for 30 minutes, and 1 μg of LPS (Lipopolysaccharide) was treated as a stimulus for 24

incubated for hours. 100 μl of the supernatant was transferred to a 96-well plate, and 100 μl of GRIESS solution was added to room temperature.

by reacting for 10 minutes and measuring the absorbance at 540 nm to determine the NO inhibitory effect of the compound of Formula 1,

The results are shown in Table 4 below.

Table 4

NO production inhibitory effect

Sample NO production inhibition rate (%)

L-NMMA (positive control, 0.001%) 42.01

Compound of Formula 1 (0.001%) 24.31

Compound of Formula 1 (0.01%) 28.23

Compound of Formula 1 (0.1%) 33.80

Compound of Formula 1 (1%) 38.18

[0098] As can be seen in Table 4, the compound of Formula 1 has a low activity compared to L-NMMA, a representative anti-inflammatory drug.

However, as it shows excellent activity as a natural substance, its anti-inflammatory effect plays a secondary role in whitening or improving wrinkles.

A synergistic effect can be expected.

Formulation Example 1: Preparation of pharmaceutical formulation

1. Preparation of powder

0.001 g of the compound of Formula 1

1 g lactose

A powder was prepared by mixing the above components and filling in airtight bags.

2. Preparation of tablets

Compound of Formula 1 0.2 mg

Corn Starch 100mg

lactose 100mg

Magnesium Stearate 2mg

After mixing the above ingredients, tablets were prepared by tableting according to a conventional tablet manufacturing method.

3. Manufacture of capsules

Compound of Formula 1 0.2 mg

Corn Starch 100mg

lactose 100mg

Magnesium Stearate 2mg

After mixing the above ingredients, capsules were prepared by filling gelatin capsules according to a conventional capsule preparation method.

4. Preparation of pills

0.003 g of the compound of Formula 1

lactose 1.5 g

1 g of glycerin

0.5 g xylitol

After mixing the above components, it was prepared to be 4 g per ring according to a conventional method.

5. Preparation of granules

2 mg of a compound of Formula 1

Soybean Extract 50 mg

Glucose 200 mg

Starch 600 mg

After mixing the above components, 100 mg of 30% ethanol was added and dried at 60 ° C to form granules, and then

Filled.

Formulation Example 2: Manufacturing of cosmetics

1. Manufacturing of softening lotion (skin lotion)

As shown in the following composition, a softening lotion containing the compound of Formula 1 as an active ingredient was prepared according to a conventional method.

0.1% by weight of the compound of Formula 1

Beta-1,3-glucan 1.0% by weight

Butylene glycol 2.0% by weight

2.0% by weight of propylene glycol

Carboxyvinyl polymer 0.1% by weight

PEG-12 nonylphenyl ether 0.2% by weight

Polysorbate 80 0.4% by weight

Ethanol 10.0% by weight

Triethanolamine 0.1% by weight

Preservative 0.05% by weight

Pigment 0.05% by weight

Perfume 0.05% by weight

Purified water 100% by weight

2. Manufacturing of nutritional lotion (milk lotion)

As shown in the following composition, a nutrient lotion containing the compound of Formula 1 as an active ingredient was prepared according to a conventional method.

0.1% by weight of the compound of Formula 1

Beta-1,3-glucan 1.0% by weight

4.0% by weight of beeswax

Polysorbate 60 1.5% by weight

Sorbitan sesquioleate 1.5% by weight

0.5% by weight of liquid paraffin

Caprylic/capric triglyceride 5.0% by weight

Glycerin 3.0% by weight

Butylene glycol 3.0% by weight

Propylene glycol 3.0% by weight

Carboxyvinyl polymer 0.1% by weight

Triethanolamine 0.2% by weight

Preservative 0.05% by weight

Pigment 0.05% by weight

Perfume 0.05% by weight

Purified water 100% by weight

3. Manufacture of nutritional cream

As shown in the following composition, a nourishing cream containing the compound of Formula 1 as an active ingredient was prepared according to a conventional method.

0.2% by weight of the compound of Formula 1

Beta-1,3-glucan 5.0% by weight

10.0% by weight of beeswax

Polysorbate 60 1.5% by weight

PEG 60 hydrogenated castor oil 2.0% by weight

Sorbitan sesquioleate 0.5% by weight

10.0% by weight of liquid paraffin

Squalane 5.0% by weight

Caprylic/capric triglyceride 5.0% by weight

Glycerin 5.0% by weight

Butylene glycol 3.0% by weight

Propylene glycol 3.0% by weight

Triethanolamine 0.2% by weight

Preservative 0.05% by weight

Pigment 0.05% by weight

Perfume 0.05% by weight

Purified water 100% by weight

4. Manufacture of massage cream

As shown in the following composition, a massage cream containing the compound of Formula 1 as an active ingredient was prepared according to a conventional method.

0.1% by weight of the compound of Formula 1

Beta-1,3-glucan 3.0% by weight

10.0% by weight of beeswax

Polysorbate 60 1.5% by weight

PEG 60 hydrogenated castor oil 2.0% by weight

Sorbitan sesquioleate 0.8% by weight

40.0% by weight of liquid paraffin

Squalane 5.0% by weight

Caprylic/Capric Triglyceride 4.0% by weight

Glycerin 5.0% by weight

Butylene glycol 3.0% by weight

Propylene glycol 3.0% by weight

Triethanolamine 0.2% by weight

Preservative 0.05% by weight

Pigment 0.05% by weight

[0198] Perfume 0.05% by weight

Purified water 100% by weight

5. Manufacture of packs

As shown in the following composition, a pack containing the compound of Formula 1 as an active ingredient was prepared according to a conventional method.

0.2% by weight of the compound of Formula 1

Beta-1,3-glucan 1.0% by weight

Polyvinyl alcohol 13.0% by weight

Sodium carboxymethyl cellulose 0.2% by weight

Glycerin 5.0% by weight

Allantoin 0.1% by weight

Ethanol 6.0% by weight

PEG-12 nonylphenyl ether 0.3% by weight

Polysorbate 60 0.3% by weight

Preservative 0.05% by weight

Pigment 0.05% by weight

Perfume 0.05% by weight

Purified water 100% by weight

Formulation Example 3: Preparation of external skin preparation

1. Preparation of the gel

As shown in the following composition, a gel containing the compound of Formula 1 as an active ingredient was prepared according to a conventional method.

0.1% by weight of the compound of Formula 1

Beta-1,3-glucan 0.1% by weight

Sodium ethylenediamine acetate 0.05% by weight

Glycerin 5.0% by weight

Carboxyvinyl polymer 0.3% by weight

Ethanol 5.0% by weight

PEG-60 hydrogenated castor oil 0.5% by weight

Triethanolamine 0.3% by weight

Preservative 0.05% by weight

Pigment 0.05% by weight

Perfume 0.05% by weight

Purified water 100% by weight

2. Preparation of ointment

As shown in the following composition, an ointment containing the compound of Formula 1 as an active ingredient was prepared according to a conventional method. 0.5% by weight of the compound of Formula 1

Beta-1,3-glucan 10.0% by weight

10.0% by weight of beeswax

Polysorbate 60 5.0% by weight

PEG 60 hydrogenated castor oil 2.0% by weight

Sorbitan sesquioleate 0.5% by weight

Vaseline 5.0% by weight

10.0% by weight of liquid paraffin

Squalane 5.0% by weight

Shea butter 3.0% by weight

5.0% by weight of caprylic/capric triglycerides

Glycerin 10.0% by weight

Propylene glycol 10.2% by weight

Triethanolamine 0.2% by weight

Preservative 0.05% by weight

Pigment 0.05% by weight

Perfume 0.05% by weight

100% by weight of purified water

3. Preparation of drugs for topical administration (gel ointment)

As shown in the following composition, a gel ointment containing the compound of Formula 1 as an active ingredient was prepared according to a conventional method.

0.5% by weight of the compound of Formula 1

Beta-1,3-glucan 10.0% by weight

Polyacrylic acid (Carbopol 940) 1.5% by weight

5.0% by weight isopropanol

Hexylene glycol 25.0% by weight

Triethanolamine 1.7% by weight

100% by weight of deionized water

4. Preparation of drugs for topical administration (patches)

As shown in the following composition, a patch containing the compound of Formula 1 as an active ingredient was prepared according to a conventional method.

0.5% by weight of the compound of Formula 1

Beta-1,3-glucan 3.0% by weight

Hexylene glycol 20.0% by weight

Diethylamine 0.7% by weight

Polyacrylic acid (Carbopol 934P) 1.0% by weight

Sodium sulfite 0.1% by weight

Polyoxyethylene lauryl ether (E.O = 9) 1.0% by weight

1.0% by weight of polyhydroxyethylene cetylstearyl ether (Cetomacrogol 1000)

2.5% by weight viscous paraffinic oil

Caprylic ester/capric acid ester (Cetiol LC) 2.5% by weight

Polyethylene glycol 400 3.0% by weight

100% by weight of deionized water

Formulation Example 4: Preparation of food

Foods containing the compound of Formula 1 of the present invention were prepared as follows.

1. Manufacture of flour food

0.05 to 1.0 parts by weight of the compound of Formula 1 is added to flour, and the mixture is used to make bread, cakes, cookies, and cakes.

Lacquer and noodles were prepared to prepare health food.

2. Manufacture of dairy products

0.2 parts by weight of the compound of Formula 1 was added to milk, and various products such as butter and ice cream were used using the milk.

A dairy product was prepared.

3. Manufacture of wire

Brown rice, barley, glutinous rice, and adlay are alphanized by a known method, dried, roasted, and then ground into powder with a particle size of 60 mesh.

was made with Black soybeans, black sesame seeds, and perilla seeds are also steamed and dried by a known method, roasted, and grinded to a particle size of 60 mesh.

It was prepared from the powder of The compound of Formula 1 was concentrated under reduced pressure in a vacuum concentrator, and dried by spraying and hot air dryer.

The obtained dried material was pulverized to a particle size of 60 mesh with a grinder to obtain a dry powder.

The following ratio with respect to 100 parts by weight of the mixed powder of the grains, seeds and dry powder of the compound of Formula 1 prepared above

It was prepared by blending at a rate.

Cereals (brown rice 30 parts by weight, adlay 15 parts by weight, barley 20 parts by weight),

Seeds (7 parts by weight of perilla seeds, 8 parts by weight of black beans, 7 parts by weight of black sesame seeds),

A compound of Formula 1 (0.1 parts by weight);

Reishiji (0.5 parts by weight),

Rehmannia glutinosa (0.5 parts by weight)

Formulation Example 5: Preparation of beverage

1. Manufacture of health drinks

Compound of Formula 1 0.1 mg

Citric Acid 1000 mg

100 g of oligosaccharides

2 g plum concentrate

1 g of taurine

Add purified water to total 900 mL

After mixing the above ingredients according to the usual health drink manufacturing method, stirring and heating at 85 ° C. for about 1 hour,

The resulting solution was filtered and obtained in a sterilized 2L container, sealed and sterilized, and then refrigerated, and then the health drink composition of the present invention

used for manufacturing.

Although the above composition ratio is a mixture of ingredients suitable for relatively favorite beverages in a preferred embodiment, the demand class, the country of demand,

It is okay to modify the mixing ratio arbitrarily according to regional and ethnic preferences such as usage.

2. Manufacture of vegetable juice

Vegetable juice for health promotion was prepared by adding 1 g of the compound of Formula 1 of the present invention to 1,000 mL of tomato or carrot juice.

all.

3. Manufacture of fruit juice

1 g of the compound of Formula 1 was added to 1,000 mL of apple or grape juice to prepare fruit juice for health promotion.

Claims (6)

A pharmaceutical composition for skin regeneration or wrinkle improvement comprising a compound represented by Formula 1 as an active ingredient:
[Formula 1] A cosmetic composition for skin regeneration or wrinkle improvement comprising a compound represented by Formula 1 as an active ingredient:
[Formula 1] An anti-inflammatory pharmaceutical composition comprising a compound represented by Formula 1 as an active ingredient:
[Formula 1] An anti-inflammatory cosmetic composition comprising a compound represented by Formula 1 as an active ingredient:
[Formula 1] A pharmaceutical composition for skin whitening comprising a compound represented by Formula 1 below as an active ingredient:
[Formula 1] A cosmetic composition for skin whitening comprising a compound represented by Formula 1 as an active ingredient:
[Formula 1]