TW201934114A - Use of Lactobacillus plantarum GMNL-6 composition for skin care capable of improving skin condition or aesthetic appearance - Google Patents

Abstract

The present invention provides a use of probiotic for preparing a skin care composition, wherein the probiotic includes Lactobacillus plantarum GMNL-6, and the depository number is BCRC 910777 or CCTCC M 2017765. The composition can be a liniment for external use, a medicine for external use, a skin care product for external use, or a cosmetic. The composition is provided with an effect of improving skin condition or aesthetic appearance by facilitating collagen secretion or expression of ceramide synthase. In addition, the Lactobacillus plantarum GMNL-6 is a dead bacterium obtained by treating a bacterial solution under high temperature and high pressure. The topical skin care product includes a toner, a lotion, a skin protective cream, a moisturizing liquid, a lip balm, a beauty mask and a facial cleanser. The composition may further include pharmaceutically and cosmetologically acceptable carriers.

Description

Use of Lactobacillus plantarum GMNL-6 composition for skin health care

The invention relates to a Lactobacillus isolate and is used in the technical field of skin care.

At present, the market emphasizes that the use of probiotics for skin is quite limited. In retrospective literature, there are two ways that probiotics affect skin characterization. One is to directly apply probiotics to the affected area, which can affect the local immune response or directly inhibit the growth of skin bad bacteria. Second, after oral probiotics enter the intestine, it affects the intestinal flora and immune response, and then affects changes in the distal skin ( Jeong JH et al , 2016 ) . For example, oral probiotics can improve or prevent heterotopic Dermatitis (Atopic Dermatitis AD) has been widely used. In retrospective studies (Baquerizo Nole KL et al , 2014, Fuchs-Tarlovsky V et al , 2016, Jeong JH et al , 2016), it was also found that some lactic acid bacteria use the acetic acid, lactic acid, butanedione, and sphingomyelin produced by them. Phospholipase, hyaluronic acid or its surface substances ... etc, can achieve the effect of promoting skin health. In general, dermal probiotics have several main effects: for example: improving acne (Kim J, Ko Y et al, 2010), the ability to resist UV damage (Ra J et al, 2014), and promoting wounds Healing effect (Peral MC et al , 2009), anti-melanin formation effect (Chan CF et al , 2014).

In the past, there were few literatures on clinical human trials using probiotics to improve skin health, and most of them were mainly oral. Studies on the formulation of probiotics added to the skin for external use were relatively rare. The resulting emulsion has a significant effect on moisturizing sensitive skin after 29 days of wiping (N = 66 persons, female) (Gueniche A et al, 2010), while clinical reports on the application of lactobacillus are still lacking.

The main function of collagen in the skin is to provide skin with complete moisture supplement, increase skin elasticity and surface gloss. Nervamine is a lipid that naturally exists in the intercellular space of the skin stratum corneum, and is the most important and highest proportion of the skin stratum corneum lipid. Its main function is to closely maintain the lipid integrity of the stratum corneum, prevent the evaporation of water, and resist foreign bacteria and injuries. The present invention mainly uses in vitro mode to screen probiotics that stimulate human skin fibroblasts to secrete and produce collagen and synthesize neural crestamine, and further uses skin testing equipment to confirm the health promotion effect of probiotic emulsions on the skin.

Since the previous research on probiotics in the field of skin care is not extensive, it is urgent to find effective probiotics for improvement. Since collagen and neural crestamine have been widely used to explore the skin state and its mechanism of action, this experiment is to use collagen and neural crestamine to explore the effects of probiotics on skin health improvement. Plant milk that may have improved skin condition has been identified Lactobacillus plantarum GMNL-6); further, the skin parameters of the test subjects were detected with a high-order VISIA digital skin texture detector.

The purpose of the present invention is to provide a use of a probiotic composition for skin health care, wherein the probiotic comprises Lactobacillus plantarum GMNL-6, and its registration number is BCRC 910777 or CCTCC M2017765.

In order to achieve the aforementioned object of the invention, the Lactobacillus plantarum GMNL-6 is a dead cell.

In order to achieve the foregoing object of the present invention, the dead bacteria are obtained by treating the bacteria-containing liquid by high temperature and high pressure.

In order to achieve the foregoing object of the invention, the composition is an external rub, an external medicine, an external skin care product or a cosmetic.

To achieve the foregoing object of the invention, the composition may further comprise a pharmaceutically or cosmetically acceptable carrier.

In order to achieve the foregoing object of the present invention, the external care product is a lotion, an emulsion, a skin cream, a moisturizing liquid, a lip balm, a cosmetic liquid mask or a facial cleanser.

To achieve the foregoing object of the invention, the skin care refers to improving skin condition or aesthetic appearance.

In order to achieve the foregoing object of the present invention, the skin condition is improved by promoting collagen secretion or expression of neural crestamine synthetase.

In order to achieve the foregoing object of the invention, the skin care system is to reduce the number of skin spots, pores, wrinkles, ultraviolet spots, or purple matter.

In order to achieve the foregoing object of the invention, the improvement of human skin condition or aesthetic appearance includes: (1) treating, reducing, and / or preventing fine lines or wrinkles; (2) reducing the size of skin pores; (3) improving skin thickness and fullness Degree, and / or firmness; (4) improve skin softness and / or softness; (5) improve skin tone, gloss, and / or clarity; (6) increase procollagen and / or collagen production; (7) Improve the maintenance and remodeling of elastin; (8) Improve skin texture and / or promote reformation; (9) Improve skin barrier repair and / or function; (10) Improve skin contour appearance; (11) Restore skin Gloss and / or brightness; (12) Supplement essential nutrients and / or components in the skin; (13) Improve skin appearance due to menopause; (14) Improve skin moisturization; (15) Increase skin elasticity and / Or elasticity; (16) treatment, reduction, And / or prevent skin sagging; (17) reduce pigmentation spots; (18) treat, reduce, and / or prevent acne or acne; or any combination thereof.

In order to achieve the foregoing object of the invention, the composition contains 1 × 10 ⁷ to 1 × 10 ¹¹ dead GMNL-6 dead cells per gram.

To achieve the foregoing object of the present invention, the number of preferred dead bacteria per gram in the composition is 1 × 10 ⁸ to 1 × 10 ¹⁰ .

Figure 1A Protein performance of collagen

Figure 1B Gene expression of Neuraminamine synthase

Figure 2 Analysis of the number of spots on the subject with the VISIIA Advanced Digital Skin Detector

Figure 3 Analysis of the number of pores in the subject with the VISIIA advanced digital skin texture detector

Figure 4 Analysis of the number of wrinkles in the subject with a VISIIA high-order digital skin texture detector

Figure 5 Analysis of the number of ultraviolet spots on the subject with the VISIIA Advanced Digital Skin Detector

Figure 6 Analysis of the amount of purple matter in the test subject with the VISIA high-order digital skin texture detector

Unless defined otherwise, all technical and scientific terms described in this specification have the meaning commonly understood by those skilled in the art.

The purpose of the present invention is to provide a use of a probiotic composition for skin health care, wherein the probiotic comprises Lactobacillus plantarum GMNL-6, and its registration number is BCRC 910777 or CCTCC M2017765. In addition, the present invention also relates to the novel use of the composition or the lactobacillus for skin health care, and its action mechanism is to promote collagen secretion and neural crestamine expression to achieve the effect of skin health care.

The Lactobacillus isolate also includes its subcultured progeny, or mutants, but still has the same characteristics, genomics, or uses (for inhibiting enterovirus) as those described in the present invention.

The compositions described herein may include, but are not limited to: food, beverages, health foods, animal water additives, animal feed additives, animal and human medical compositions, food additives, beverage additives, etc. Its application form.

The term "pharmaceutically or cosmetically acceptable carrier" means that the substance or composition must be compatible with the other ingredients of the formulation and not harmful to the patient.

The term "Cell Lysate" may be a single extract obtained from a particular extraction step or series of extraction steps, or the extract may be a combination of several extracts obtained from several independent extraction steps. These combined extracts are therefore also encompassed by the term "extract".

The pharmaceutical or cosmetically acceptable carrier may include one or more agents selected from the group consisting of a solvent, an emulsifier, a suspending agent, a decomposer, and a binder ( binding agent), excipient, stabilizing agent, chelating agent, diluent, gelling agent, preservative, lubricant , Surfactants, and other carriers similar or applicable to the present invention.

To the above composition, one or more of dissolution aids, buffers, preservatives, colorants, flavors, flavors, and the like, which are generally used in the field of preparations, may also be appropriately added as needed.

In addition, the novel strains discovered in the present invention can also be included in a composition together with other conventional strains.

The composition provided by the present invention and its use for skin health care, the dosage form thereof can be appropriately adjusted according to requirements, and is not particularly limited, and the skin external dosage form is preferred.

The present invention is exemplified by the following examples, but the present invention is not limited by the following examples. The medicines and biological materials used in the present invention are all commercially available and easily available, and the following are merely examples of available pipelines.

The present invention is exemplified by the following examples, but the present invention is not limited by the following examples.

Select the Lactobacillus plantarum with skin health effects from the strain bank. The storage number, date and strain name of the Lactobacillus are shown in Table 1.

Example 1 GMNL-6 can promote collagen secretion and expression of neuraminamine synthase

1-1 Dead bacteria preparation steps:

Lactobacillus plantarum (GMNL-6) was inoculated with 1 ml MRS broth from a freezer tube, and cultured at 37 ° C under aerobic conditions for 20 hours. On the next day, take 15 μl of overnight culture bacterial solution to 1.5 ml MRS broth (1% secondary activation), incubate at 37 ° C for 20 hours under aerobic conditions, wash it once with PBS, estimate the bacterial count at OD600nm, and adjust the bacterial count. It was 1 × 10 ¹⁰ cells / ml and was autoclaved at 121 ° C. It was used after 15 minutes.

Human skin fibroblast (Hs68) cell model:

To analyze the collagen of the cell culture solution: seed Hs68 cells to a 6well plate (2 × 10 ⁵ cells / well), wash them twice with PBS overnight, change to serum free medium, and add 1 × 10 ⁹ cells / ml of GMNL-6 dead bacteria liquid, after 24 hours of treatment, the supernatant was collected.

The supernatant was diluted 10-fold, and then analyzed using a Procollagen type I C-peptide EIA kit (TaKaRa, # MK101).

To analyze the intracellular neuraminamine synthetase gene (Serine palmitoyltransferase small subunit A, SPTSSA) steps:

Hs68 cells were seeded into a 6well plate (2 × 10 ⁵ / well), washed overnight with PBS twice, replaced with serum free medium (medium), and added with 1 × 10 ⁹ cells / ml of dead bacteria bacteria solution, treated After 24 hours, the cell extracts were collected, RNA was extracted and transformed into cDNA, and quantitative PCR analysis of neural aminamine synthase gene (SPTSSA) and housekeeping gene (β-actin) was performed.

Experimental results:

Previously with human skin fibroblasts in vitro screening platform model, from dozens of Lactobacillus strains were screened Lactobacillus (L. plantarum) having a skin care effect of GMNL-6, its capacity to promote secretion of collagen. Furthermore, in the same mode, the experimental results (Figure 1) have found that GMNL-6 has the ability to promote collagen secretion and stimulate the production of intracellular neuraminamine synthetase (SPTSSA). Neuraminamine synthase is a nerve in the body. An essential molecule in the synthesis of scopolamine. Its rise also represents an increase in the production of scopolamine in the body. Scobamine has enhanced skin moisturization, increased skin defense, and inhibited the production of tyrosinase in skin beauty.

Example 2: Lactobacillus plantarum GMNL-6 probiotic emulsion has the function of improving the skin quality of the entire face

2-1 Experimental steps:

1. Number of trials: five women (30-40 years).

2. Evaluation place: Functional Cosmetics Development and Evaluation Research Center of Jianan Pharmaceutical University.

3. Before the test, cleanse the face with a facial cleanser, perform a full-face skin diagnosis with VISIA Advanced Digital Skin Tester, and record the value (day0)

4. During the test, apply the test sample daily after washing your face in the morning and evening (except for the lotion, the other maintenance conditions on the left and right faces are the same):

-For left half face: control lotion (matrix)

-For right half face: probiotic emulsion (matrix + GMNL-6 dead bacteria)

5. Perform a full face skin diagnosis (day 28) after four weeks of use. First clean the face with a cleanser, then perform a full face skin diagnosis, and record the value.

2-2 emulsion ingredients and probiotics concentration:

1.Emulsion ingredients (matrix): Water, Polyglyceryl-3 / Methyl glucose / Distearate, GlycerylTrioctanoin, Macadamia Ternifolia Seed Oil, Propanediol, PEG / PPG / Polybutylene Glycol-8 / 5/3 Glycerin, Cetyl Alcohol, MyristylMyristate, Glycerin, GlycosylTrehalose / Hydrogenated Starch Hydrolysate, Rice Ferment Filtrate (Sake), Saccharomyces / Daylily Flower Ferment Filtrate, Bis-PEG / PPG-20 / 15 PEG / PPG-20 / 5Dimethicone / Methoxy PEG / PPG-25 / 4 DimethiconeCaprylic / Capric Triglyceride, Sodium Polyacrylate, Phenoxyethanol, o-Cymen-5-ol, Xanthan Gum, Potassium Hydroxide.

2. GMNL-6 concentration in emulsion: 1 × 10 ⁹ cells / g emulsion

2-3 Evaluation method:

1. Use VISIA high-order digital skin texture detector to perform skin quality detection, take high-resolution photos of the face with a 12-megapixel camera and three different light sources (normal light, UV light, polarized light), and then use software to analyze the facial fixation The range of skin quality can accurately obtain skin parameters such as spots, pores, wrinkles, texture, purple spots, and purple matter to evaluate the underlying condition of the skin's basal layer. This will allow you to understand skin problems and evaluate changes in data before and after using the lotion. The test results are compared based on gender, age and a qualified normal range data, which can be used as an evaluation of the effectiveness of the product.

2.Evaluate each person's own left face (control lotion) and right face (probiotics) through the skin texture parameters (including spots, pores, wrinkles, purple spots, and purple matter) measured by the full-face skin texture diagnostic instrument. Emulsion) The difference in scores before (day0) and after (day 28). The calculation formula for the score change (%) is as follows:

Experimental results:

Five subjects (NO.1 ~ 5) used the control lotion (left face) and probiotics Lotion (right face). After 28 days, the skin parameters of five subjects were analyzed via the VISIIA Advanced Digital Skin Detector, including spots (Figure 2), pores (Figure 3), wrinkles (Figure 4), and purple spots (Figure 4). Figure 5) and Violet (Figure 6). After comparing each person with their left face, it is estimated that using the probiotic emulsion group can get better skin quality improvement data. At least three people in each evaluation item have improved, especially the spots and Pores, as much as 80 to 100% of people, have significantly improved. Therefore, in general, probiotics have the effect of improving skin texture, especially in reducing spots and reducing pores.

The collagen and neural crestamine in the skin will gradually disappear with the increase of age or bad living habits. Therefore, supplement and restore the collagen and neural crestamine content in the skin, which is good for skin health, anti-wrinkle and anti-UV. Damage and anti-aging are very important. The evaluation of this patent found that Lactobacillus plantarum GMNL-6 can promote collagen secretion and expression of neural crestamine synthetase at the same time, which represents that GMNL-6 has special effects on the skin. Observed by human clinical experiments via the VISIIA high-order digital skin texture detector, Lactobacillus plantarum GMNL-6 probiotic emulsion has the function of improving the skin quality of the entire face, including: spots, pores, wrinkles, purple spots, and purple matter, each of which has at least three people The above improvement is especially effective for people who dilute spots and reduce pores, up to 80 ~ 100%; In addition, the decrease of purple matter means that the growth of acne bacteria is inhibited, which should be helpful for improving acne and acne. The clinical application of probiotics in skin and human body is mostly oral, and the clinical trials of directly adding dead lactic acid bacteria to the emulsion are few, and the evaluation method that compares their left and right faces at the same time is used to reduce the interference of other maintenance products and other individual differences. Cause of change. And five people who used GMNL-6 probiotic emulsion did not show any allergies and discomfort after four weeks of use. Therefore, the use of Lactobacillus plantarum (GMNL-6) in skin care products is quite safe and has no side effects. It should be a better choice to promote skin health and prevent skin aging in the future.

The above-mentioned multiple effects are actually in line with the statutory invention patent requirements of novelty and advancement. I applied for it in accordance with the law, and I urge your office to approve this invention patent application to encourage invention.

[Biological Material Storage]

Domestic Deposit Information [Please note according to the order of deposit organization, date, and number]

Hsinchu Food Industry Development Institute Biological Resources Preservation and Research Center, the deposit date is April 14, 2017, and the deposit number is BCRC 910777.

Information on foreign deposits [Please note according to the order of the depositing country, institution, date, and number]

China, deposit date is November 3, 2017, deposit number is CCTCC M 2017765

Claims (12)

A use of a probiotic for preparing a skin health composition, wherein the probiotic comprises Lactobacillus plantarum GMNL-6, and its registration number is BCRC 910777 or CCTCC M2017765. The use as described in item 1 of the patent application scope, wherein the Lactobacillus plantarum GMNL-6 is a dead bacterium. The use as described in item 1 of the scope of patent application, wherein the dead bacteria body is obtained by treating the bacteria-containing liquid by high temperature and high pressure. The use as described in item 1 of the scope of the patent application, wherein the composition is an external rub, an external medicine, an external care product or a cosmetic. The use as described in item 1 of the patent application scope, wherein the composition may further comprise a pharmaceutically or cosmetically acceptable carrier. The use as described in item 1 of the scope of the patent application, wherein the external care product is a lotion, lotion, skin cream, moisturizer, lip balm, facial mask or facial cleanser. The use as described in item 1 of the scope of patent application, wherein the skin care refers to improving skin condition or aesthetic appearance. The use as described in item 7 of the scope of the patent application, wherein the improvement of skin condition is through promotion of collagen secretion or expression of neural crestamine synthetase. The use according to item 1 of the scope of patent application, wherein the skin care system is to reduce the number of skin spots, pores, wrinkles, ultraviolet spots or purplish. The use according to item 1 of the scope of patent application, wherein the improvement of human skin condition or aesthetic appearance includes: (1) treating, reducing, and / or preventing fine lines or wrinkles; (2) reducing the size of skin pores; (3) ) Improve skin thickness, fullness, and / or firmness; (4) improve skin softness and / or Softness; (5) Improve skin tone, gloss and / or transparency; (6) Increase procollagen and / or collagen production; (7) Improve elastin maintenance and remodeling; (8) Improve skin texture and / Or promote reformation; (9) improve skin barrier repair and / or function; (10) improve skin contour appearance; (11) restore skin gloss and / or brightness; (12) supplement essential skin nutrients and / or Components; (13) improving skin appearance due to menopause; (14) improving skin moisturizing; (15) increasing skin elasticity and / or elasticity; (16) treating, reducing, and / or preventing skin sagging; ( 17) reducing pigmentation spots; (18) treating, reducing, and / or preventing acne or acne; or any combination thereof. The use as described in item 1 of the scope of patent application, wherein each gram of the composition contains 1 × 10 ⁷ to 1 × 10 ¹¹ dead GMNL-6 bacterial cells. The use according to item 1 of the scope of patent application, wherein the number of better dead bacteria per gram in the composition is 1 × 10 ⁸ to 1 × 10 ¹⁰ .