COSMETIC COMPOSITION INCLUDING GARLIC EXTRACT, BLACK TEA EXTRACT, AND GUAVA EXTRACT

Abstract

Provided is a cosmetic composition which can ameliorate wrinkles and suppress secretion of sweat and sebum, thereby exhibiting a makeup lasting effect.

Claims

1. A cosmetic composition for improving a skin condition, comprising, as an active ingredient, a garlic extract, a black tea extract, and a guava extract.

2. The cosmetic composition of claim 1, wherein the garlic extract, the black tea extract, and the guava extract are mixed at a weight ratio in a range of about 1:0.1:0.1 to about 1:10:10.

3. The cosmetic composition of claim 1, wherein the garlic extract, the black tea extract, or the guava extract is obtained by performing an extraction process on garlic, black tea, or guava by using, as a solvent, water, a C1-C4 lower alcohol, or a mixture thereof.

4. The cosmetic composition of claim 1, wherein the skin condition is selected from skin elasticity amelioration, reduction in fine line appearance, wrinkle amelioration, eye-widening effect, lifting of the corners of the mouth, reduction in muscle mass, and flattening of line unfolding from the upper lip.

5. The cosmetic composition of claim 1, further comprising a guava leaf extract, a Luffa cylindrica extract, and a water lily flower extract.

6. The cosmetic composition of claim 1, wherein the cosmetic composition is for suppressing secretion of sweat or sebum or for improving or enhancing makeup durability.

7. A pharmaceutical composition for preventing or treating neuromuscular-related diseases, comprising, as an active ingredient, a garlic extract, a black tea extract, and a guava extract.

8. The pharmaceutical composition of claim 7, wherein the neuromuscular-related disease is selected from muscle tension, muscle spasm, hemifacial spasm, adult onset spasmodic torticollis, anal fissure, blepharospasm, facial muscle spasm, cerebral palsy, headache, migraine, muscle pain, strabismus, temporomandibular joint disorder, nerve pain, overactive bladder, urgency urinary incontinence, rhinitis, paranasal sinusitis, acne, pore enlargement, dystonia, dystonic contraction, hyperhidrosis, vocal cord dysfunction, and myocardial damage.

9. A cosmetic composition for improving a skin condition, comprising myricetin as an active ingredient.

10. The cosmetic composition of claim 9, wherein myricetin is derived from: chemical synthesis; biosynthesis using a microorganism; or a plant or a mushroom selected from broccoli, cauliflower, cabbage, Chinese cabbage, kalian, green chili, red chili, bell pepper, bird chili, Chinese chives, onion, garlic, belimbi, yam, tapioca, sweet potato, papaya, cashew, fern, soybean, mungbean, okra, winged bean, French bean, pea, string bean, petai, peria, brinjal, angular loofa, snake gourd, pumpkin, sengkuang, guava, carrot, white radish, red spinach, bayam duri, kangkung, wolfberry, drumstick, celery, limau purut, daun turi, betel, pandan, lemon, semambu, kesom, Maman (Gynandropsis gynandra), Kadok, cekur manis, selom, pegaga, bunga kantan, plantain, asam gelugor, turmeric, mint, oyster mushroom, and black tea.

11. The cosmetic composition of claim 9, wherein the skin condition is selected from skin elasticity amelioration, reduction in fine line appearance, wrinkle amelioration, eye-widening effect, lifting of the corners of the mouth, reduction in muscle mass, and flattening of line unfolding from the upper lip.

12. The cosmetic composition of claim 9, wherein the cosmetic composition is for suppressing secretion of sweat or sebum or for improving or enhancing of makeup durability.

13. A pharmaceutical composition for preventing or treating neuromuscular-related diseases, comprising myricetin as an active ingredient.

14. The pharmaceutical composition of claim 13, wherein the neuromuscular-related disease is selected from muscle tension, muscle spasm, hemifacial spasm, adult onset spasmodic torticollis, anal fissure, blepharospasm, facial muscle spasm, cerebral palsy, headache, migraine, muscle pain, strabismus, temporomandibular joint disorder, nerve pain, overactive bladder, urgency urinary incontinence, rhinitis, paranasal sinusitis, acne, pore enlargement, dystonia, dystonic contraction, hyperhidrosis, vocal cord dysfunction, and myocardial damage.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0059] The above and other aspects, features, and advantages of certain embodiments of the disclosure will be more apparent from the following description taken in conjunction with the accompanying drawings, in which:

[0060] FIG. 1 shows a result of confirming membrane fusion rate of a composition according to an embodiment;

[0061] FIG. 2 shows a result of confirming whether or not myricetin is detected and amount of detected myricetin;

[0062] FIG. 3 shows a result of confirming the VAMP2 level by treatment with the composition or myricetin according to an embodiment;

[0063] FIG. 4 shows a result of confirming the acetylcholine level by treatment with the composition or myricetin according to an embodiment;

[0064] FIG. 5 shows a result of confirming changes in an amount of skin sebum by the composition according to an embodiment for each time period;

[0065] FIG. 6 shows a result of confirming changes in skin color by the composition according to an embodiment for each time period; and

[0066] FIG. 7 shows a result of confirming rates of skin color change by the composition according to an embodiment for each time period.

DETAILED DESCRIPTION

[0067] Reference will now be made in detail to embodiments, examples of which are illustrated in the accompanying drawings, wherein like reference numerals refer to like elements throughout. In this regard, the present embodiments may have different forms and should not be construed as being limited to the descriptions set forth herein. Accordingly, the embodiments are merely described below, by referring to the figures, to explain aspects of the present description. As used herein, the term and/or includes any and all combinations of one or more of the associated listed items. Expressions such as at least one of, when preceding a list of elements, modify the entire list of elements and do not modify the individual elements of the list.

[0068] Hereinafter, preferable Examples are presented to help understanding of the present disclosure. However, the following examples are only presented for easier understanding of the present disclosure, and the contents of the present disclosure are not limited by the following examples.

PREPARATION EXAMPLES

Preparation Example 1. Preparation of Garlic Extract

[0069] Fresh garlics grown on the farm were harvested and washed thoroughly with distilled water. After drying the garlics for a week to remove moisture, 100 g of the dried garlics was pulverized and immersed in 1,000 g of 70% ethanol (EtOH), followed by stirring and extraction at room temperature for 3 days. Afterwards, the extract was filtered by centrifugation, and the filtrate was repeatedly extracted twice in the same manner as described above. The extract from which the filtrate was removed was concentrated with a rotary vacuum concentrator to prepare a concentrate, and then freeze-dried to prepare a garlic extract.

Preparation Example 2. Preparation of Black Tea Extract

[0070] A black tea extract was prepared in the same manner as in Preparation Example 1, except that 100 g of black tea was used.

Preparation Example 3. Preparation of Guava Extract

[0071] A guava extract was prepared in the same manner as in Preparation Example 1, except that 100 g of guava was used.

Preparation Example 4. Preparation of Guava Leaf Extract

[0072] A guava leaf extract was prepared in the same manner as in Preparation Example 1, except that 100 g of guava leaf was used.

Preparation Example 5. Preparation of Luffa cylindrica Fruit Extract

[0073] A Luffa cylindrica fruit extract was prepared in the same manner as in Preparation Example 1, except that 100 g of Luffa cylindrica fruit was used.

Preparation Example 6. Preparation of Water Lily Flower Extract

[0074] A water lily flower extract was prepared in the same manner as in Preparation Example 1, except that 100 g of water lily flower was used.

Examples

Example 1. Preparation of Compositions Including Garlic Extract, Black Tea Extract, and Guava Extract

[0075] A cosmetic composition was prepared by mixing each of the extracts prepared in Preparation Examples 1 to 3 in the same amount (33.3 wt %). Unless otherwise indicated in the present specification, an amount is expressed in wt %.

Example 2. Preparation of Composition Including Garlic Extract, Black Tea Extract, Guava Extract, Guava Leaf Extract, Luffa cylindrica Fruit Extract, and Water Lily Flower Extract

[0076] A cosmetic composition was prepared by mixing 1.5 wt % of each of the extracts prepared in Preparation Examples 1 to 3, 1.0 wt % of each of the extracts prepared in Preparation Examples 4 and 5, 3.0 wt % of the extract prepared in Preparation Example 6, 2.0 wt % of an adjuvant (e.g., 1,2-hexanediol), 30.0 wt % of butylene glycol, and 58.5 wt % of purified water.

Example 3. Preparation of Freeze-Dried Product of Composition Including Garlic Extract, Black Tea Extract, Guava Extract, Guava Leaf Extract, Luffa cylindrica Fruit Extract, and Water Lily Flower Extract

[0077] A freeze-dried product of the composition prepared in Example 2 was prepared by freeze-drying. The freeze-dried product had a yield of about 500 mg/L.

Comparative Examples

Comparative Examples 1 to 3. Preparation of Composition Including Extract

[0078] A garlic extract, a black tea extract, and a guava extract of Comparative Examples 1 to 3 were prepared in the same manner as in Preparation Examples 1 to 3.

Experimental Examples

Experimental Example 1. Confirmation of Inhibitory Effect on Membrane Fusion

[0079] The inhibitory effect of the composition according to an embodiment on the membrane fusion was confirmed. Also, since myricetin was predicted as an active ingredient, the inhibitory effect of 100 M of myricetin (Sigma-Aldrich) on the membrane fusion was also confirmed.

[0080] First, a soluble nethylmaleimide-sensitive factor attachment protein (SNAP) receptor protein was overexpressed in transformed E. coli, and then the expressed protein was selectively purified and obtained by using glutathione agarose beads. Then, 1-palmitoyl-1,2-dioleoyl-sn-glycero-3-phosphatidylcholine (POPC, 62 mol %), 1,2-dioleoyl-sn-glycero-3-phosphatidylserine (DOPS, 35 mol %), 1,2-dioleoyl-sn-glycero-3-phosphoserine-N-(7-nitro-2-1,3-benzoxadiazol-4-yl) (NBD-PS, 1.5 mol %), and a fluorescent material, rhodamine (rhodamin-PE, 1.5 mol %) were mixed to prepare a 10 mM fluorescent substance-labeled liposome (v-vesicle). Meanwhile, DOPS and POPC were mixed to a molar concentration of 35:65 to prepare a 50 mM liposome (t-vesicle) that is not labeled with a fluorescent substance. Then, purified synaptosomal-associated protein, SNAP-25, and syntaxin 1a were mixed at a molar concentration of 1:1 and allowed for a reaction for 1 hour at room temperature, followed by mixing with the liposome that is not labeled with a fluorescent substance at a molar ratio of 100:1. Also, vesicle-associated membrane protein 2 (VAMP-2) was mixed with the fluorescent substance-labeled liposome at a molar ratio of 50:1. These two types of the liposomes were dialyzed and mixed at a molar ratio of 1:9 (v-vesicle:t-vesicle), and treated with 100 g/ml of each of the extracts of Example 1 and Comparative Examples 1 to 3.

[0081] Then, the fluorescence intensity thereof was measured.

[0082] FIG. 1 shows a result of confirming the membrane fusion rate by the treatment with the composition and myricetin according to an embodiment.

[0083] As a result, as shown in FIG. 1, the composition of Example 1 including all of the garlic extract, the black tea extract, and the guava extract showed a significantly reduced membrane fusion rate compared to the extracts of Comparative Examples 1 to 3 each containing the extract of single component, and the membrane fusion rate was most significantly reduced by myricetin.

[0084] That is, the composition according to an embodiment inhibits the formation of SNARE complex by acting specifically on the SNARE protein, and thus may exhibit a synergistic activity on the inhibition of membrane fusion. Also, it was confirmed that the inhibition effect on membrane fusion may be due to myricetin.

[0085] Accordingly, the composition not only inhibits the secretion of sweat and/or sebum by blocking the release of neurotransmitters and inhibits the generation of wrinkles by musculation or sympathetic and parasympathetic nerves, but also ameliorates wrinkles already generated. Such effects may be exhibited also due to the myricetin.

Experimental Example 2. Confirmation of Myricetin Detection

[0086] It was confirmed whether myricetin included in the composition according to an embodiment was detected.

[0087] In detail, the composition of Example 3 was dissolved in the same volume of methanol (MeOH) as the original composition, filtered through a 0.45-g PVDF filter, and then quantified by using the reverse-phase HPLC system with a UV system and the INNO C18 column. Here, the column temperature was maintained at 35 C., the flow rate was maintained at 1 mL/min, the sample input was set at 10 microliter, and the measurement wavelength was set at 350 nm. For a fluidized phase, a 0.5% acetic acid aqueous solution (A) and acetonitrile (B) were used, and a concentration gradient was set as 80% A at 0 min, 70% A at 30 min, and 80% A at 35 min. By using a standard curve constructed using 1 ppm to 100 ppm of myricetin, the amount of myricetin included in the sample was calculated.

[0088] As a result, as shown in FIG. 2, the myricetin was detected in Example 3, and when calculated by using the standard curve, it was confirmed that the amount of myricetin in the composition of Example 3 was 13.85 mg/L.

[0089] Therefore, it was confirmed that the composition according to an embodiment contained the myricetin as an active ingredient.

Experimental Example 3. Confirmation of Effects on Inhibition of SNARE Protein and Suppression of Neurotransmitter Release in Human Skin Tissue

[0090] First, the inhibitory effect of the composition and myricetin according to an embodiment on the SNARE protein was confirmed. In detail, the human skin explant, which has been disinfected with povidone and completely underwent fat removal, was evenly cut with an 8-mm biopsy punch, and then cultured for 24 hours in a transwell to which 1.5 mL medium (DMEM+10% FBS+10 g/mL insulin+10 ng/mL hydrocortisone+2 mM L-glutamine) was added. 10 L of each of the composition of Example 3 and the myricetin sample was treated evenly over the entire skin at the respective set concentrations, and then further cultured for 72 hours. As a positive control, botulinum toxin type A (botox cosmetics) was used. The cultured tissues have been completely cultured were fixed with formalin for fluorescence staining with VAMP2 (ab215721) which is one of the SNARE proteins. The fluorescence-stained tissues were photographed under a confocal microscope, and the fluorescence intensity of VAMP2 was analyzed by using Image J.

[0091] Next, the inhibitory effect of the composition and myricetin according to an embodiment on the release of neurotransmitters was confirmed. In detail, the human skin explant, which has been disinfected with povidone and completely underwent fat removal, was evenly cut with an 8-mm biopsy punch, and then cultured for 24 hours in a transwell to which 1.5 mL medium (DMEM+10% FBS+10 g/mL insulin+10 ng/mL hydrocortisone+2 mM L-glutamine) was added. 10 L of each of the composition of Example 3 and the myricetin sample was treated every day for 3 days at the respective set concentrations. As a positive control, botulinum toxin type A (botox cosmetics) was used. The cultured tissues were stored in LN.sub.2. After uniformly homogenizing the tissues stored in LN.sub.2, proteins were quantified by using the Bradford assay. Each sample was adjusted to the same amount of the proteins, and the amount of a neurotransmitter, acetylcholine, was measured at Ex/Em 535/587 nm by using the choline/acetylcholine assay kit (ab65345).

[0092] FIG. 3 shows a result of confirming the VAMP2 level by treatment with the composition or myricetin according to an embodiment.

[0093] FIG. 4 shows a result of confirming the acetylcholine level by treatment with the composition or myricetin according to an embodiment.

[0094] As a result, as shown in FIGS. 3 and 4, the VAMP2 level and the amount of acetylcholine released in the skin tissue were significantly reduced by the composition of Example 3 or the myricetin.

[0095] That is, the composition according to an embodiment inhibited the formation of SNARE complex by acting specifically on the SNARE protein and accordingly inhibited the membrane fusion so that the release of neurotransmitters was inhibited. It was confirmed that such effects may be exhibited by the myricetin.

[0096] Accordingly, the composition not only inhibited the secretion of sweat and/or sebum by blocking the release of neurotransmitters and inhibited the generation of wrinkles by musculation or sympathetic and parasympathetic nerves, but also ameliorated wrinkles already generated. It was confirmed that such effects may be exhibited by the myricetin.

Experimental Example 4. Confirmation of Inhibitory Effect on Sebum Secretion

[0097] The inhibitory effect of the composition according to an embodiment on the sebum secretion was confirmed.

[0098] In detail, the composition of Example 2 was applied to one side of the face of 22 women in their 20s to 50s to be absorbed, and then a reference color sample (Dr. Jart+ The Makeup Fit Cushion 01 Light product) was applied thereto (excluding the forehead). Thereafter, after washing the face with an existing cleanser, the face was lightly patted with a paper towel to remove moisture, and then rested for 30 minutes under constant temperature and humidity conditions (20 C. to 24 C., 40% to 60% RH). After the face of a test subject was photographed, sites for the measurement were divided, and the amount of sebum secreted on the skin was measured 5 times each by using a sebumeter before the application, immediately after the application, and 3 hours, 6 hours, and 8 hours after the application. The average value of the measured values is shown in FIG. 5.

[0099] FIG. 5 shows a result of confirming changes in the amount of skin sebum by the composition according to an embodiment for each time period.

[0100] As a result, as shown in FIG. 5, it was confirmed that the amount of sebum secreted was 56.77 g/cm.sup.2 immediately after the application, 116.27 g/cm.sup.2 after 3 hours of the application, 140.86 g/cm.sup.2 after 6 hours of the application, and 174.64 g/cm.sup.2 after 8 hours of the application, in the case where the composition of Example 2 was not applied. Meanwhile, in the case where the composition of Example 2 was applied, it was confirmed that the amount of sebum secreted was 40.59 g/cm.sup.2 immediately after the application, 70.91 g/cm.sup.2 after 3 hours of the application, 95.32 g/cm.sup.2 after 6 hours of the application, and 115.27 g/cm.sup.2 after 8 hours of the application, and accordingly that there was a statistically significant difference from immediately after the start of the test after 8 hours.

[0101] That is, when the composition of Example 2 was applied, it was confirmed that the amount of sebum secreted was low at a statistically significant level (p<0.05) until 8 hours of the application, compared to the case where the same composition was not applied.

[0102] Therefore, the composition according to an embodiment was able to decrease or inhibit the sebum secretion, which may be also due to myricetin as an active ingredient.

Experimental Example 5. Confirmation of Makeup Durability Effect

[0103] The effect of the composition according to an embodiment on the makeup durability was confirmed.

[0104] In detail, the composition of Example 2 was applied to one side of the face of 22 women in their 20s to 50s to be absorbed. After washing the face with an existing cleanser, the face was lightly patted with a paper towel to remove moisture, and then rested for 30 minutes under constant temperature and humidity conditions (20 C. to 24 C., 40% to 60% RH). Next, a reference color sample (Dr. Jart+ The Makeup Fit Cushion 01 Light product) was applied to the testing site (except for the forehead), and the face of the test subject was photographed immediately after the application and after 3 hours, 6 hours, and 8 hours of the application. Here, the changed in the skin color (L-value) of the test subject was measured by using a chromameter.

[0105] FIGS. 6 and 7 show a result of confirming changes and rates of skin color change by the composition according to an embodiment for each time period.

[0106] As a result, as shown in FIGS. 6 and 7, in both cases where the composition of Example 2 was applied or not applied, the measured values for the skin color before and after the application showed the same pattern. After the application of the standard color sample, it was confirmed that the skin color changed at a statistically significant level (p<0.05) by the makeup effect of the colored cosmetics. However, compared to the case where the composition of Example 2 was not applied, the skin color in the case where the same composition was applied increased at a high level, and it was confirmed that such an increase was statistically significant from 3 hours after application to 8 hours after application.

[0107] That is, the composition according to an embodiment not only exhibited an effect on the skin color improvement by color makeup, but also improved makeup durability, and such effects may be due to myricetin which is an active ingredient.

[0108] According to the one or more embodiments, a composition including a garlic extract, a black tea extract, and a guava extract, or a composition including myricetin may inhibit membrane fusion by the formation of SNARE complex and inhibit the release of neurotransmitters, so that the wrinkles may be ameliorated and the secretion of sweat and sebum may be inhibited, thereby exhibiting effects of makeup durability and preventing or treating neuromuscular-related diseases.

[0109] It should be understood that embodiments described herein should be considered in a descriptive sense only and not for purposes of limitation. Descriptions of features or aspects within each embodiment should typically be considered as available for other similar features or aspects in other embodiments. While one or more embodiments have been described with reference to the figures, it will be understood by those of ordinary skill in the art that various changes in form and details may be made therein without departing from the spirit and scope of the disclosure as defined by the following claims.