KR20070102910A - A cosmetic composition preventing a formation of snare complex containing a extract of magnoliae cortx - Google Patents

Abstract

A cosmetic composition containing the extract of Magnoliae Cortex is provided to prevent the formation of SNARE(soluble NSF(N-ethylmaleimide-sensitive factor) attachment protein receptor) complex which is associated with the neurotransmission pathway by functioning at neuromuscular junction, thereby blocking secretion of acetylcholine, inducing paralysis of the terminal motor nerve and inhibiting repetitive contraction and relaxation of skin muscle. A cosmetic composition for improving skin wrinkles contains 0.005-50 wt.% of the extract of Magnoliae Cortex which is prepared by extracting Magnoliae Cortex with a solvent selected from water, C1-C4 lower alcohol and a mixed solvent thereof, inhibits binding of Syntaxin and SNAP-25 and is formulated as skin lotion, skin softener, astringent, lotion, milk lotion, moisture lotion, nutrition lotion, massage cream, nutrition cream, moisture cream, hand cream, essence, nutrition essence, pack, soap, shampoo, cleansing foam, cleansing lotion, cleansing cream, body lotion, body cleanser, emulsion, press powder, loose powder or eye shadow.

Description

A cosmetic composition preventing a formation of SNARE complex containing a extract of Magnoliae Cortx}

1 is a result confirming the inhibitory effect on the formation of snare (SNARE) complex of the present invention.

The present invention relates to a composition for inhibiting snare complex formation containing a glucobacterium extract.

In other words, the present invention is a acetylcholine extract extract acts at the neuromuscular junction (Neuromuscular junction) to interfere with the formation of snares (recognition receptors present on the surface of the vesicle involved in SNARE intracellular transport) complex acetyl By blocking the secretion of choline, it has the effect of paralyzing the extremity of the motor nerve, thereby suppressing the repetitive contraction and relaxation of skin muscles.

In addition, the cosmetic composition containing the extract of Danghubak Park having such an effect can be utilized as a cosmetic composition for preventing skin aging, such as skin wrinkle improvement effect, collagen synthesis promoting effect.

Cosmetics are deeply involved in our daily lives and used by many people. Before the development of human civilization, the purpose of cremation was to protect the body from nature.However, with the development of science and civilization, the purpose of cremation is to keep the body clean and beautiful, and to delay the aging to make a beautiful and pleasant life. Changed. The function of cosmetics is not only clean and aesthetic, but also free radicals and free radicals caused by external factors such as moisture, pollutants and stress, ultraviolet rays, and internal factors such as deterioration of immune cells and deterioration of cell activity. This is to remove or suppress the cause of skin aging such as protein, nucleic acid, and cell membrane lipid destruction by peroxides. In addition, cosmetics having whitening, skin wrinkle improvement and suppression, and sunscreen functions to meet the consumer's desire for high-performance cosmetics continue to be developed. In particular, wrinkle improvement and suppression products are of great interest to consumers seeking to stay younger.

Wrinkles are formed by repetitive movements of muscles in a specific direction for a long time, affected by age, external environment, and UV irradiation. Damage to skin cells caused by these factors causes various aging phenomena. The collagen, elastin, and GAGs constituting the dermal matrix are not easily produced and degraded, resulting in poor firmness and wrinkles of the skin.

Currently, wrinkle improvement cosmetics are known as retinoids, adenosine, animal placental protein, chlorella extract and the like. Retinol, the most well known, promotes collagen synthesis and inhibits elastase enzymes, but it is unstable and has limited use due to safety issues such as irritation and seizures when applied to the skin. Chlorella extract is practically a wrinkle It is difficult to expect improvement effects.

Botulinum toxin called botox has been used for the purpose of wrinkle improvement through muscle paralysis. Botox is a brand name for muscle contraction injections of Allergan Inc., a US pharmaceutical company that is used to remove nerve and muscle diseases and wrinkles, and is made by purifying botulinum toxin. Botulinum toxin is a toxin made by a bacterium called clostridium botulinum that occurs mainly in spoiled canned foods. Injecting Botox into your muscles will slow the movement of your muscles for a period of time by blocking the neurotransmitters that move your muscles. A more detailed mechanism of action is that botulinum toxin acts at the neuromuscular junction, which interferes with the formation of SNARE (recognition receptors present on the surface of the medial receptor involved in intracellular transport) complex acetyl Blocking the secretion of choline causes flaccid paralysis, showing its effect. The mechanism by which action occurs consists of three steps, first by the specific interaction of the heavy chain with cell surface receptors, the toxin binds to the presynaptic membrane of the target neuron. The toxin then passes across the plasma membrane of the poisoned cell. Finally, the light chain chain selectively cleaves proteins necessary for the recognition and neurotransmitter containing vesicles and the cytoplasmic surface of the plasma membrane and for talking and fusion to cause degradation of membrane proteins.

On the other hand, it is not known about the use of Danghubak extract to suppress snare complex formation.

It is an object of the present invention to inhibit the repetitive contraction and relaxation of skin muscles by inhibiting the end of the motor nerve by blocking the secretion of acetylcholine by interfering with the formation of snare complex involved in the neurotransmission pathway containing herbal extracts This is to show the effect.

As a result of researching various herbal extracts in order to achieve this technical problem, it was found that the extract of Danghubak Park has the effect of inhibiting the formation of the snare complex, and to provide it to the cosmetic composition to improve the skin condition for the purpose of the present invention. have.

The present invention provides a composition for inhibiting the formation of snare complex (SNARE) containing the extract.

The glucobacterium extract is characterized in that the polar solvent selected from water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof, most preferably, is hydrothermally extracted by adding ethanol.

In addition, the composition is characterized in that it is utilized to improve the skin condition by inhibiting the formation of the snare complex as a cosmetic.

Magnolia officinalis Rehd. Et Wils. Is a deciduous arbor tree, about 10 meters tall, bark grayish brown with hairs on the surface of young branches. The leaves are rosacea, elliptical obovate, and the ends of the leaves are circular. The plant is relatively large in its leaves, different from other plants, and grows almost straight upright. The tree is unique and exotic, so it is often used as a garden or tree. Yellow-green flowers bloom in May-June, fruits are in September-October, fruits are large and black.

The herbal medicine called hoobak is called ganhubak ( Magnolia Cortex) in China. In Korea, there is a herbal medicine called Hanfubak , which is used for the same purpose as the Machili cortex of the bark of Machilus thunbergil, which belongs to the camphor tree. The main ingredients are β-eudesmol (machilol), α, β-pinene, camphene, limonene, bornylacetate, caryophyllene, epoxide, magnolol, honokiol, neolignans, α-eudesmol, cryptomeridiol, magnocurarine, magnoflorine, anonaine, michelarbine, liriodenine, salici to be.

Danghubak extract according to the present invention can be prepared as follows.

The green pepper was washed thoroughly with water and dried in the shade. The dried sugar flakes are placed in an extraction container, and extracted for a predetermined time at an appropriate temperature in an appropriate amount of extraction solvent. The extract can be filtered and concentrated using a vacuum rotary evaporator or the like.

As the extraction solvent, a polar solvent selected from water (purified water), a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof may be used, and preferably ethanol is used.

The amount of the extraction solvent is 2 to 10 times the dry weight of the herbal medicine.

The extraction method may be an extraction method such as water bath extraction, reflux cooling extraction, ultrasonic extraction, etc., preferably may be extracted 1 to 5 times as an extraction method of water bath extraction.

The deposition temperature is 30 ~ 70 ℃, more preferably 40 ~ 60 ℃, most preferably maintained at about 50 ℃ is most effective in the prevention of decay, discoloration and odor.

The deposition time is 5 to 20 hours, preferably 10 hours.

After obtaining the glucobacterium extract, the solids may be removed using a filter paper, the suspension may be centrifuged, and the supernatant may be filtered under reduced pressure.

The filtered extract can be concentrated under reduced pressure at 20 ~ 100 ℃, preferably 50 ~ 70 ℃ with a vacuum rotary concentrator.

Danghubak extract according to the present invention inhibits the formation of snare complexes, indicating the use of skin condition improvement. The cosmetic composition of the present invention improves wrinkles of the skin by inhibiting snare complex formation inhibitory effect and cell growth promoting effect, which is not toxic compared to Botox, and has excellent stability in the formulation. This fact is specified in the following examples.

In the composition of the present invention, the sanghakbak extract may be contained 0.005 to 50% by weight based on the total weight of the composition.

The composition of the present invention may further contain at least one active ingredient exhibiting the same or similar function in addition to the sugar pak extract.

Cosmetics prepared with a cosmetic composition containing the extract of Danghubak as an active ingredient can be prepared in the form of a general emulsion formulation and solubilization formulation. Cosmetics of the emulsion formulations include nutrient cosmetics, creams, essences, and the like. Cosmetics in the formulation include soft cosmetics. In addition to the cosmetic containing the extract of Danghubak Park of the present invention by containing a dermatologically acceptable medium or base may be prepared in the form of adjuvants that can be applied topically or systemically applied commonly used in the field of dermatology.

Suitable cosmetic formulations include, for example, emulsions, suspensions, microemulsions, microcapsules, microgranules or ionics (liposomes), nonionics obtained by dispersing an oil phase in a solution, gel, solid or pasty anhydrous product, aqueous phase. It may be provided in the form of a vesicle dispersant, in the form of a cream, skin, lotion, powder, ointment, spray or conceal stick. It may also be prepared in the form of a foam or in the form of an aerosol composition further containing a compressed propellant.

In addition, the cosmetic composition of the present invention, in addition to sugar extract, fatty substances, organic solvents, solubilizers, thickening and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, Commonly used in water, ionic or nonionic emulsifiers, fillers, metal ion sequestrants and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or cosmetics And any other ingredients that may be used, such as auxiliaries commonly used in the cosmetic or dermatological arts. In addition, the above components may be introduced in an amount generally used in the dermatology field.

Examples of products to which the cosmetic composition of the present invention may be added include, for example, cosmetics such as astringent cosmetics, soft cosmetics, nourishing cosmetics, various creams, essences, packs, foundations, cleansing agents, face washes, soaps, treatments, Essences and the like.

Specific formulations of the cosmetic composition of the present invention include skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisturizing lotion, nutrition lotion, massage cream, nutrition cream, moisture cream, hand cream, essence, nutrition essence, pack, Formulations such as soaps, shampoos, cleansing foams, cleansing lotions, cleansing creams, body lotions, body cleansers, emulsions, press powders, loose powders, eye shadows and the like.

Hereinafter, preferred examples and experimental examples are presented to help understand the present invention. However, the following Examples and Experimental Examples are provided only to more easily understand the present invention, and the contents of the present invention are not limited by the Examples.

Preparation Example 1 Preparation of Sugar Gourd Extract

50 g of Danghubak was finely crushed in a crusher and immersed in 200 ml of 100% ethanol and warmed in a 50 ° C. water bath for 10 hours. The obtained extract was cooled to room temperature and filtered through a filter paper to give a filtrate. Ethanol was evaporated from the filtrate using nitrogen, concentrated, and then dissolved in 10% ethanol to prepare an extract.

Experimental Example 1 Inhibition Effect of Snare Complex Formation

In this experiment, the effect of inhibiting the formation of the snare complex in the signal transduction of neurons was measured by immunoprecipitation. Neurotransmitters are mediated by the regulation of exocytosis of synaptic vesicles. The snare complex is involved in neuronal exocytosis and consists of the vesicle protein VAMP (synaptobrevin) and the plasma membrane binding protein syntaxin 1 and Snap-25 (SNAP-25).

In this experiment, to determine the inhibition of binding to Syntaxin 1 and Snap-25 (SNAP-25) by Danghubak extract, we synthesized a snare complex protein from PC-12 cells and synthesized Syntaxin 1 After immunoprecipitation using an antibody against the purified protein, the precipitated protein was purified, and then Western blotting was performed using an antibody against syntaxin 1 and snap-25 (SNAP-25). It was.

PC-12 cells (Korea Cell Line Bank) were inoculated at 3 × 10 ⁶ cells / well in T75 flasks containing DMEM medium (sigma-aldrich) containing 10% bovine serum and 5% CO ₂ at 37 ° C for 24 hours. Incubate. After removing the medium and replacing with a fresh medium, the sugar gourd extract prepared in Preparation Example 1 was added at an appropriate concentration, followed by incubation for 48 hours to recover each cell. Wash cells twice with PBS, homogenize 50 times with 23-gauge needle in homogenization buffer containing complete protease inhibitor mixture (Roche): 23mga needle, 150mM NaCl, 2.5mM KCl, 10mM Na2HPO4, pH 7.4 Centrifugation at rpm for 10 minutes yielded plasma membrane pellets. The pellet was dissolved in 1% Triton X-100, 20mM HEPES-KOH, pH 7.2, 2mM EDTA, 100mM KCl, 1mM DTT containing a complete protease inhibitor mixture and reacted at 4 ° C for 10 minutes, followed by centrifugation at 13,000 rpm for 20 minutes. The supernatant was taken off.

200 μg of the protein was reacted with antibody (monoclonal antibodies against syntaxin 1 (Gttingen) or Snap-25 (SNAP-25, Gttingen) for 30 minutes at 4 ° C.) Protein A-Separose (Amersham) was added and 4 The reaction was shaken gently for 12 hours at 0 ° C. The beads were precipitated by centrifugation and washed four times with digestion buffer, SDS-sample buffer was added to the precipitate, and SDS-PAGE and immunoprecipitation were performed by standard methods. Immobilon-P membrane (Millipore) was used for electroblotting and horseradish peroxidase-conjugated IgG was used as a secondary antibody.

As shown in FIG. 1, it was found that addition of glucobacterium extract was inhibited in the composition of the snare complex, that is, the binding of syntaxin 1 and snap-25 (SNAP-25). This inhibitory mechanism was interpreted to be due to the ability of Danghubak extract to bind to syntaxin 1 competitively with Snap-25 (SNAP-25).

Experimental Example 2 Cell Proliferation Promoting Effect

This experiment measured the cell proliferation promoting effect of Danghubak extract. Danghubak extract was dissolved in an appropriate concentration in a buffer solution and used as a sample solution. Mouse-derived NIH / 3T3 fibroblasts (Korea Cell Line Bank) were inoculated in DMEM medium containing 4.5 g / L glucose, 10% serum, and 1% antibiotics and cultured at 37 ° C. in a 75 cm 2 T-flask. After incubation for 24 hours in 5% CO ₂ and treated with 0.02% trypsin to recover the cells were inoculated with 2X10 ³ cells / well in a 96well plate and incubated for 6 hours. The sample solution of appropriate concentration was diluted in DMEM medium, treated with cultured fibroblasts, and cultured at 37 ° C for 4 days. After incubation, a portion of the medium is removed and the reaction is performed for 4 hours by adding MTT solution. The MTT solution reacts in the mitochondria of living cells to form yellow to purple formazan crystals. After removing the sample, DMSO solution was added to dissolve the formazan crystal, and the absorbance was measured at 570 nm to compare the cell proliferation.

Final experimental concentration Absorbance (570nm) 0 0.952 25 μg / ml 0.956 50 μg / ml 1.103 100 μg / ml 1.255 200 μg / ml 1.549

As can be seen in the table, the extract of Danghubak Park was shown to promote the proliferation of cells depending on the concentration.

Preparation Example 2 Examples and Comparative Examples

To prepare a cosmetic containing the extract of Danghukbak of Preparation Example 1, in order to compare it with a cosmetic composition lacking a dougakbak extract was prepared in the composition ratio of Table 2.

division Content (% by weight) ingredient Example Comparative example Moroccan Extract 1.0 - Butylene glycol 2.0 2.0 Propylene glycol 6.0 6.0 Carboxy Vinyl Polymer 0.5 0.5 Beeswax 2.0 2.0 vaseline 7.0 7.0 Polysorbate 60 2.0 2.0 Sorbitan sesquioleate 2.5 2.5 Squalane 3.0 3.0 Liquid paraffin 10.0 10.0 Triethanolamine 0.5 0.5 Tocopheryl Acetate 0.1 0.1 Spices, preservatives a very small amount a very small amount Purified water Remaining amount Remaining amount Sum 100 100

Experimental Example 3 Clinical Effect of External Wrinkle Improvement

Clinical effects of skin wrinkle improvement on cosmetics containing Danghubak extract of the present invention were measured. Twenty women aged 40 years or older (mean age 44.5 years) were divided into two groups (A and B), and an example for group A and a comparative example for group B were prepared according to Table 2, respectively. After 12 weeks of application (2 times / day, 0.2 g / time), the improvement of skin wrinkles (ΔW) was measured by classifying the subject's objective evaluation and subject's subjective evaluation into the following 7 levels. . The results are shown in Tables 3 and 4 below.

In addition, the skin wrinkle effect (Skin Visiometer SV 600 FireWire, CK Electronic GmbH) and the elasticity measuring instrument (Cutometer MPA 580, CK Electronic GmbH) were used to measure wrinkle improvement and elasticity of the skin. Is shown in Table 5.

* Skin wrinkle evaluation criteria

Deteriorate very badly -3 worse -2 Slightly worse -One No change 0 Slightly improved One Improving 2 Very improved 3

Objective evaluation of the skilled person Average wrinkle improvement (W) Example Comparative example T0 T12 T0 T12 Subject 1 0 2 0 One Subject 2 0 3 0 0 Subject 3 0 One 0 0 Subject 4 0 2 0 0 Subject 5 0 3 0 0 Subject 6 0 2 0 One Subject 7 0 One 0 0 Subject 8 0 2 0 0 Subject 9 0 3 0 0 Subject 10 0 3 0 One W (T12-T0) 2.2 0.3

Subjective Evaluation of the Subject division Average wrinkle improvement (W) Example Comparative example T0 T12 T0 T12 Subject 1 0 One 0 One Subject 2 0 3 0 0 Subject 3 0 2 0 0 Subject 4 0 3 0 0 Subject 5 0 One 0 One Subject 6 0 3 0 0 Subject 7 0 2 0 0 Subject 8 0 3 0 One Subject 9 0 2 0 0 Subject 10 0 2 0 One W (T12-T0) 2.2 0.4

As a result of the test, after 12 weeks, the Examples containing the Danghubak extract was found to have a significant wrinkle improvement effect as the average wrinkle improvement of the subjects' objective evaluation and the subject's subjective evaluation of 2.2, respectively.

division Wrinkle improvement effect Elasticity Average depth of wrinkles (%) Roughness average (%) Skin adsorption degree (%) Example T0 100.0 100.0 100.0 T12 75.5 62.4 66.5 Comparative example T0 100.0 100.0 100.0 T12 99.1 96.4 97.5

In Table 5, the average depth of the wrinkles is divided into five equal parts of the sample made of silicone rubber at the site where wrinkles are to be measured, and represents the average value of the difference between the highest and lowest points of the wrinkle height in each of five places. The roughness mean represents the mean value of the difference in wrinkle depth at each measurement site (dividing the whole into 255 measurement site points) from the average value of the total wrinkle depth in the sample. The lower the average depth and roughness average of the wrinkles, the smaller the wrinkles are formed, which means that the degree of skin aging is small.

      From the results shown in Table 5, when looking through the relative numerical calculation before / after the application of the embodiment, the wrinkles improved by 22.1% after the application of the example when the 100% before the application of the average depth value of wrinkles, and looked at the roughness average Since the effect was found to be 37.4% improved, it can be seen that the cosmetic composition containing Danghubak extract according to the present invention has a considerable effect on the improvement of wrinkles of the skin.

       In addition, the skin adsorption degree in Table 5 represents the degree of adsorption as the skin depth (the penetration depth of the skin into the aperture) into the measuring apparatus, the smaller the value is preferable. When viewed as 100% before application, it can be confirmed that the elasticity of the skin is improved to a significant extent by 32.23% after the application of the above example.

      As a result, it can be confirmed through the results shown in Table 5 that the cosmetics containing the herbal extract prepared by the extraction example according to the present invention to improve the skin wrinkle effect and elasticity effect, excellent effect of preventing skin aging It can be seen that.

Formulation Example 1 Flexible Lotion

     Based on the results of the previous experimental example, to present a cosmetic composition containing the sugar extract of the present invention. However, the composition of the present invention is not intended to be limited to the following formulation examples. Formulation examples of the flexible lotion containing the sanghakbak extract was prepared as shown in Table 6.

Raw material Content (% by weight) Moroccan Extract 1.00 1,3-butylene glycol 1.00 Disodium ID 0.05 Allantoin 0.10 Dipotassium glycylizate 0.05 Citrix Acid 0.01 Sodium citrate 0.02 Glyceres-26 1.00 Arbutin 2.00 Hydrogenated Castor Oil 1.00 ethanol 30.00 Preservative a very small amount coloring agent a very small amount Flavor a very small amount Purified water Remaining amount

Formulation Example 2 Nutrition Cream

Formulation example of the nutrient cream containing the extract of Danghubak was prepared as shown in Table 7.

Raw material Content (% by weight) Moroccan Extract 2.0 1,3-butylene glycol 7.0 glycerin 1.0 D-panthenol 0.1 Plant extracts 3.2 Magnesium Aluminum Silicate 0.3 PEG-40 Stearate 1.2 Stearic acid 2.0 Polysorbate 60 1.5 Lipophilic glyceryl stearate 2.0 Sorbitan sesquioleate 1.5 Cetearyl Alcohol 3.0 Mineral oil 4.0 Squalane 3.8 Carlylic / Capric Triglycerides 2.8 vegetable oil 1.8 Dimethicone 0.4 Dipotassium glycylizate a very small amount Allantoin a very small amount Sodium hyaluronate a very small amount Tocopheryl Acetate Quantity Triethanolamine Quantity Preservative Quantity Flavor Quantity Purified water Remaining amount

As described above, the present invention exhibits the effect of paralyzing the ends of the motor nerve by blocking the secretion of acetylcholine through the composition for inhibiting the formation of snare complexes containing a glucobacterium extract, thereby repeatedly contracting and relaxing the skin muscle. It shows the effect of suppressing. When the composition containing the extract of Danghubak Park is used in the cosmetic composition, it is useful for cosmetic compositions for preventing skin aging, such as wrinkle improvement effect and collagen synthesis promoting effect.

Claims (8)

Snare complex containing the extract for snare (SNARE) complex formation. The composition of claim 1, wherein the composition inhibits the binding of syntaxin 1 and snap-25 (SNAP-25). The composition of claim 1, wherein the extract is Magnoliae Cortex . The composition of claim 1, wherein the extract is hydrothermally extracted. The composition of claim 4, wherein the hydrothermal extraction is performed by adding a polar solvent selected from water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. A cosmetic composition for improving skin wrinkles, comprising the composition of any one of claims 1 to 5. 7. The cosmetic composition according to claim 6, wherein the content of the saccharide extract is 0.005 to 50 wt% based on the total weight of the cosmetic composition. According to claim 6, wherein the cosmetic composition is a skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisturizing lotion, nutrition lotion, massage cream, nutrition cream, moisture cream, hand cream, essence, nutrition essence, pack Cosmetic composition, characterized in that the selection of any one selected from the group consisting of soap, shampoo, cleansing foam, cleansing lotion, cleansing cream, body lotion, body cleanser, emulsion, press powder, loose powder and eye shadow.

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