External preparation for skin

Abstract

To provide an external preparation for skin containing a plant-derived lactic acid bacterium and a specified plant extract.SOLUTION: An external preparation for skin comprises bacterial cells of Lactobacillus casei subsp. casei strain 327 and a specified plant extract.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to an external skin preparation containing plant-derived lactic acid bacteria and a specified plant extract, and more specifically to an external skin preparation containing Lactobacillus casei subsp. casei strain 327 cells and a specified plant extract. [Background technology]

[0002] The present applicants have confirmed that adding cells of Lactobacillus casei subsp. casei strain 327 makes it possible to maintain skin moisture (Patent Document 1). [Prior art documents] [Patent documents]

[0003] [Patent Document 1] Patent No. 6805399 Summary of the Invention [Problem to be solved by the invention]

[0004] Therefore, the present inventors have newly confirmed the synergistic effect of combining Lactobacillus casei subsp. casei strain 327 cells with a specific plant extract. [Means for solving the problem]

[0005] Specifically, the means are as follows: (1) A topical skin preparation containing cells of Lactobacillus casei subsp. casei strain 327 deposited under accession number NITE BP-02707 and Melissa extract. (2) A topical skin preparation containing cells of Lactobacillus casei subsp. casei strain 327 deposited under accession number NITE BP-02707 and an aloe vera extract. (3) The agent according to (1) or (2), wherein the bacterial cells are killed bacterial cells. (4) An agent described in any one of (1) to (3), which is for promoting the growth of Staphylococcus epidermidis. DETAILED DESCRIPTION OF THE INVENTION

[0006] The present disclosure will be described below using examples, but the present disclosure is not limited to the following examples and can be implemented with any modifications. Furthermore, the descriptions in this disclosure are mutually applicable unless otherwise specified. In this specification, the expression "to" is used to include the numerical or physical values ​​before and after it. In this specification, the expression "A and / or B" includes "A only," "B only," and "both A and B."

[0007] (Plant lactic acid bacteria) The plant-based lactic acid bacteria used in the present invention are isolated from plant-derived sources, including processed foods such as miso paste, soy sauce, pickles, bran, grass, rice, wheat, and malt, and are capable of producing lactic acid by utilizing carbohydrates. The plant-based lactic acid bacteria are preferably Lactobacillus lactic acid bacteria isolated from fermented foods made from rice. Several plant-based lactic acid bacteria isolated from rice and processed rice products have been reported to have antimutagenic properties ( Journal of the Japanese Society for Food Science and Technology, Vol. 48, No. 9, pp. 693-696, 2001). Among these, the plant-based lactic acid bacteria used as an active ingredient in the present invention are most preferably Lactobacillus casei subsp. casei 327 or its mutant strains. Here, the term "mutant" refers to a specific strain mutated by a method well known to those skilled in the art without altering its properties, or a strain that can be confirmed by those skilled in the art to be equivalent to such a strain.

[0008] Lactobacillus casei subsp. casei strain 327 was deposited domestically at the National Institute of Technology and Evaluation, Patent Microorganisms Depositary (Room 122, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, 292-0818, Japan) on May 8, 2018 (deposit date). The deposit number is NITE BP-02707. (Hereinafter, this strain will be referred to as "strain K-1").

[0009] The lactic acid bacteria used as an active ingredient in the present invention may be either live or killed. However, killed bacteria are preferred, for example, to prevent the generation of unpleasant odors due to the proliferation of live bacteria. The lactic acid bacteria are more preferably heat-sterilized, obtained by sterilizing the lactic acid bacteria using a known heat treatment method. Heat-sterilized cells can be prepared by culturing the lactic acid bacteria in MRS medium (Difco) or similar in a conventional manner, recovering the cells by filtration, centrifugation, or other methods, washing them with water, suspending them in water, or the like, and heat-treating them at 120°C or less (preferably 80 to 120°C) for 30 minutes or less (3 seconds to 30 minutes), followed by concentration and drying, as necessary. Alternatively, the cells may be baked or steamed (for example, at 170°C or less for 60 minutes or less). The K-1 strain of lactic acid bacteria is available from the Food Development Institute of Kameda Seika Co., Ltd., under the name "Plant-derived Lactic Acid Bacteria K-1." Therefore, in the present invention, such commercially available products may be used as the lactic acid bacteria used as an active ingredient.

[0010] (External skin preparations and their effects) The "external skin preparation" of the present invention is a preparation or composition having a skin care effect, containing the above-mentioned K-1 strain fungus cells and a predetermined plant extract as active ingredients. Here, "skin care" typically refers to strengthening or restoring the skin's natural function as a barrier against environmental influences such as dust, chemicals, and microorganisms, and against the loss of endogenous substances such as water, natural fats, and electrolytes in skin tissue. Furthermore, the killed fungus cells used in a preferred embodiment are not only expected to have the above-mentioned skin care effects, but also can be used preferably because they do not undergo any further morphological changes, whereas live fungus cells may undergo morphological changes during delivery or display after product production.

[0011] (Promoting the growth of Staphylococcus epidermidis, etc.) Staphylococcus epidermidis, a type of resident skin flora, has an antagonistic effect against pathogenic bacteria such as Staphylococcus aureus and can inhibit the growth of such harmful bacteria. Therefore, the ability to promote the growth of Staphylococcus epidermidis is thought to be one function of a balance adjuster for the resident skin flora (Japanese Patent Publication No. 2021-195341). A "balance improver for resident skin flora" is a formulation or composition that maintains the resident skin flora present on healthy skin and provides a barrier function that prevents the invasion of pathogenic bacteria from outside. When the balance of the resident skin flora is disrupted, excessive proliferation of resident bacteria occurs, and the invasion and proliferation of other harmful bacteria occurs, inducing various skin symptoms. In the present invention, promotion of the growth of Staphylococcus epidermidis can be achieved, for example, by improving the balance of resident skin flora (increasing the proportion of Staphylococcus epidermidis in the resident skin flora, and thereby expected to improve skin quality, such as moisturizing).

[0012] (Plant extract) Plant extracts (also called plant extracts) are solutions and their dried products extracted from plants using various extraction methods. The components dissolved in the solvent (solute) form the solute, and the combination of the solvent and solute forms the solution (see Cosmetic Ingredients Online, Plant Extract Explanation and Ingredient List). Plant extracts have a long history of use in folk medicines and cosmetics, both as medicinal herbs in the West and as traditional Japanese herbal medicines in the East. Some of these extracts are also approved as pharmaceuticals in Japan, and are known to have a variety of beneficial effects depending on the plant species, extraction part, and extraction method. Medicinal herbs and traditional Japanese herbal medicines are formulated to exert synergistic or combined effects in addition to their individual effects. Therefore, in cosmetics, mixed ingredients designed to exert synergistic or combined effects when combined are commonly used.

[0013] (Melissa extract) The melissa extract is an extract of the leaves of Melissa officinalis Linne (Labiatae). The melissa extract is prepared, for example, by extracting the leaves of Melissa officinalis L. (Labiatae) with a 1,3-butylene glycol solution. The melissa extract used in the following examples is Falcorex Melissa B (manufactured by Ichimaru Falcos Co., Ltd. (a product produced by adding a 1,3-butylene glycol solution to the leaves of Melissa officinalis L. (Labiatae) and extracting, followed by cold treatment, adjustment, and filtration)).

[0014] (Aloe vera extract) The aloe vera extract is an extract obtained from the sap of the leaves of Aloe barbadensis Miller (Aloe vera Linne) or Aloe arborescens Mill. (Liliaceae) or its variants. If necessary, aloin and other components may be removed from this extract. The aloe vera extract used in the following examples is Falcorex Aloe KB (manufactured by Ichimaru Falcos Co., Ltd. (a product obtained by adding 1,3-butylene glycol solution to the leaves of Aloe arborescens Mill. (Liliaceae) and its variants for extraction, followed by filtration, cold treatment, conditioning, and filtration). [Example]

[0015] Next, examples of the present disclosure will be described. However, the present disclosure is not limited by the following examples. Commercially available reagents were used according to their protocols unless otherwise specified. Note that "mol / l" may also be abbreviated as "M." In the following examples, the unit % in numerical values ​​means % by mass.

[0016] [Experiment 1] Experiment using a culture system of Staphylococcus epidermidis To screen for substances that promote the growth of S. epidermidis in vitro, a culture system for S. epidermidis was established. The media and bacterial strains used for this co-culture were as follows: NB liquid medium (Nutrient Broth No. 2) was prepared by dissolving NB medium (Kanto Chemical Co., Ltd., 711067-5 Nutrient Broth No. 2) in water and then sterilizing it in an autoclave. The dissolution was carried out at a ratio of 25 g of NB medium to 1000 ml of water. NB agar medium is made by adding agar to NB medium to make it 1.5% and solidifying it. Staphylococcus epidermidis (S. epidermidis): ATCC12228 Plant extract: Falcorex Aloe KB (manufactured by Ichimaru Falcos Co., Ltd.)

[0017] (Culture preparation) A frozen stock of the bacteria to be used was prepared in advance. This frozen stock was appropriately diluted with NB liquid medium and plated on an agar medium. The viable bacterial concentration in the original stock was calculated from the number of colonies. 9 CFU / mL frozen stocks were obtained. Next, NB liquid medium was added to a 96-well plate, followed by the samples to be used in the test. In this experiment 1, the following samples were added as samples to be used in the test. Sample addition group: Killed cells of Lactobacillus casei subsp. casei strain 327 (killed lactic acid bacteria) at a final concentration of 0.05% and Falcorex Aloe KB (plant extract) at a final concentration of 1% were added to make the final volume 100 μL. Control group: Instead of killed lactic acid bacteria and plant extract, an aqueous solution containing the same amount of 1,3-butylene glycol as the corresponding sample was used.

[0018] (culture) A frozen stock of Staphylococcus epidermidis was thawed and cooled on ice. In this experiment, the final concentration of Staphylococcus epidermidis was 5 × 10 6 The 96-well plate containing Staphylococcus epidermidis was gently shaken to mix, and then cultured at 37°C for 20 hours.

[0019] (Plating of culture medium) After 20 hours of incubation, the 96-well plate was removed from 37°C and cooled on ice. The S. epidermidis culture was diluted with pre-prepared, autoclaved PBS (Fujifilm Wako Pure Chemical Industries, Ltd., 162-19321, phosphate-buffered saline powder). An appropriate amount of the diluted culture was added dropwise to NB agar medium, spread with a Conlarger stick as needed, and incubated overnight at 37°C.

[0020] (Results tabulation) After overnight culture, the number of colonies grown on the NB agar medium was counted. The results are shown in Table 1 below. The counting was performed three times for each group (each addition group). The values ​​shown in Table 1 below represent the average values ​​(relative values, actual measured values) of the results of the three counting operations.

[0021] [Table 1]

[0022] [Experiment 2] Experiment using a culture system of Staphylococcus epidermidis To screen for substances that promote the growth of S. epidermidis in vitro, a culture system for S. epidermidis was established. The media and strains used for this culture are as follows: NB liquid medium (Nutrient Broth No. 2) was prepared by dissolving NB medium (Kanto Chemical Co., Ltd., 711067-5 Nutrient Broth No. 2) in water and then sterilizing it in an autoclave. The dissolution was carried out at a ratio of 25 g of NB medium to 1000 ml of water. NB agar medium is made by adding agar to NB medium to make it 1.5% and solidifying it. Staphylococcus epidermidis (S. epidermidis): ATCC12228 Plant extract: Falcorex Melissa B (manufactured by Ichimaru Falcos Co., Ltd.)

[0023] (Culture preparation) A frozen stock of the bacteria to be used was prepared in advance. This frozen stock was appropriately diluted with NB liquid medium and plated on an agar medium. The viable bacterial concentration in the original stock was calculated from the number of colonies. 9 CFU / mL frozen stocks were obtained. Next, NB liquid medium was added to a 96-well plate, followed by the samples to be used in the test. In this experiment 2, the following samples were added as samples to be used in the test. Sample addition group 1: Killed cells of Lactobacillus casei subsp. casei strain 327 (killed lactic acid bacteria) at a final concentration of 0.05% and Falcorex Melissa BB (manufactured by Ichimaru Falcos Co., Ltd.) at a final concentration of 1% were added to a final volume of 100 μL. Sample addition group 2: Killed cells of Lactobacillus casei subsp. casei strain 327 (killed lactic acid bacteria) at a final concentration of 0.05% and Falcorex Melissa B (manufactured by Ichimaru Falcos Co., Ltd.) at a final concentration of 2% were added to a final volume of 100 μL. Control group: An aqueous solution prepared to contain the same amount of 1,3-butylene glycol as the corresponding sample instead of killed lactic acid bacteria cells and plant extract. Control group 1 corresponds to sample addition group 1, and control group 2 corresponds to sample addition group 2.

[0024] (culture) A frozen stock of Staphylococcus epidermidis was thawed and cooled on ice. In this experiment, the final concentration of Staphylococcus epidermidis was 5 × 10 6 The 96-well plate containing Staphylococcus epidermidis was gently shaken to mix, and then cultured at 37°C for 20 hours.

[0025] (Plating of culture medium) After 20 hours of incubation, the 96-well plate was removed from 37°C and cooled on ice. The S. epidermidis culture was diluted with pre-prepared, autoclaved PBS (Fujifilm Wako Pure Chemical Industries, Ltd., 162-19321, Phosphate Buffered Saline Powder). An appropriate amount of the diluted culture was added dropwise to NB agar medium, spread with a Conlarger as needed, and incubated overnight at 37°C.

[0026] (Results tabulation) After overnight culture, the number of colonies grown on the NB agar medium was counted. The results are shown in Table 2 below. The count was performed three times for each group (each addition group). Based on the count results, the relative values ​​of the colony counts for sample addition group 1 and sample addition group 2 were calculated, with control group 1 or control group 2 being set at 100. The values ​​shown in Table 2 below indicate the average values ​​(relative values, measured values) of the count results performed three times.

[0027] [Table 2]

[0028] The following topical skin preparations were prepared in Formulation Examples 1 to 12. The formulations of the topical skin preparations are shown below. In the following formulation examples, the lactobacillus is, for example, killed cells of the 327 strain (killed lactic acid bacteria cells).

[0029] Prescription example 1. Lotion [Ingredients] Part Jojoba oil 0.1~1.0 PEG-60 Hydrogenated Castor Oil 0.1~5.0 Phenoxyethanol 0.01~1.0 Ceramide 0.0001~1.0 Ethanol 0.01 to 10.0 Glycerin 0.01~10.0 1,3-butylene glycol 0.01~10.0 pH adjuster (appropriate amount) Lactobacillus 0.0001~0.1 Plant extract (Melissa extract or Aloe vera extract) 0.0001~2.0 Active ingredient (GK2 (dipotassium glycyrrhizinate), etc.) Amount of active ingredient Active ingredients (plant extracts, etc.) 0.0001~5.0 Purified water (enough to make the total volume 100 parts)

[0030] Prescription example 2: Emulsion [Ingredients] Part Squalane 0.1~10.0 Sunflower oil 0.1~10.0 Polyoxyethylene (20) sorbitan monostearate 0.01~5.0 Soy lecithin 0.01~2.0 Ceramide 0.0001~1.0 Glycerin 0.01~10.0 1,3-butylene glycol 0.01~10.0 Xanthan gum 0.01 to 2.0 Phenoxyethanol 0.01~1.0 pH adjuster (appropriate amount) Lactobacillus 0.0001~0.1 Plant extract (Melissa extract or Aloe vera extract) 0.0001~2.0 Active ingredient (GK2, etc.) Amount of active ingredient Active ingredients (plant extracts, etc.) 0.0001~5.0 Purified water (enough to make the total volume 100 parts)

[0031] Prescription example 4. Cream [Ingredients] Part Squalane 0.1~10.0 Triethylhexanoin 0.1~10.0 Cetyl alcohol 0.01~5.0 Decaglyceryl stearate 0.01~5.0 Glyceryl stearate 0.01~5.0 Soy lecithin 0.01~2.0 Glycerin 0.01~10.0 Ceramide 0.0001~1.0 1,3-butylene glycol 0.01~10.0 Sodium hyaluronate 0.01~2.0 Carbomer 0.2~0.4 Phenoxyethanol 0.01~1.0 pH adjuster (appropriate amount) Lactobacillus 0.0001~0.1 Plant extract (Melissa extract or Aloe vera extract) 0.0001~2.0 Active ingredient (GK2, etc.) Amount of active ingredient Active ingredients (plant extracts, etc.) 0.0001~5.0 Purified water (enough to make the total volume 100 parts)

[0032] Prescription example 5. Sunscreen [Ingredients] Part Ethylhexyl methoxycinnamate 0.01~10.0 Diethylaminohydroxybenzoylhexyl benzoate 0.01~10.0 Polyoxyethylene sorbitan monooleate (20E.O.) 0.01-5.0 Triethylhexanoin 0.1~10.0 Glycerin 0.01~10.0 1,3-butylene glycol 0.01~10.0 Phenoxyethanol 0.01~1.0 Carboxyvinyl polymer 0.01~1.5 pH adjuster (appropriate amount) Lactobacillus 0.0001~0.1 Plant extract (Melissa extract or Aloe vera extract) 0.0001~2.0 Active ingredient (GK2, etc.) Amount of active ingredient Active ingredients (plant extracts, etc.) 0.0001~5.0 Purified water (enough to make the total volume 100 parts)

[0033] Prescription example 6. Sunscreen [Ingredients] Part Polyhydroxystearic acid 0.1~40.0 Polyglyceryl-3 Polyricinoleate 0.01~5.0 Polyglyceryl monoisostearate 3 0.01~5.0 Disteardimonium Hectorite 2 0.01~5.0 Titanium oxide 0.1~20.0 Zinc oxide 0.1~20.0 Hydroxylated AL 0.01~10.0 Isostearic acid 0.01~10.0 1,3-butylene glycol 0.1~10.0 Phenoxyethanol 0.01~1.0 pH adjuster (appropriate amount) Lactobacillus 0.0001~0.1 Plant extract (Melissa extract or Aloe vera extract) 0.0001~2.0 Active ingredient (GK2, etc.) Amount of active ingredient Active ingredients (plant extracts, etc.) 0.0001~5.0 Purified water (enough to make the total volume 100 parts)

[0034] Prescription example 7. Balm [Ingredients] Part Polyglyceryl-2 triisostearate 0.1~10.0 Beeswax 0.1~10.0 Carnauba wax 0.1~10.0 1,3-butylene glycol 0.1~5.0 Lactobacillus 0.0001~0.1 Plant extract (Melissa extract or Aloe vera extract) 0.0001~2.0 Active ingredient (GK2, etc.) Amount of active ingredient Active ingredients (plant extracts, etc.) 0.0001~5.0 Squalane Amount that makes the total amount 100 parts

[0035] Formulation example 8. Liquid foundation [Ingredients] Part Triethylxanoin 0.1~40.0 Polyglyceryl-3 Polyricinoleate 0.1~10.0 Polyglyceryl monoisostearate 3 0.1~10.0 Disteardimonium Hectorite 2 0.1~10.0 1,3-butylene glycol 0.1~10.0 Phenoxyethanol 0.01~1.0 Titanium oxide 0.1~10.0 Mica 0.1~10.0 Color pigment (appropriate amount) Lactobacillus 0.0001~0.1 Plant extract (Melissa extract or Aloe vera extract) 0.0001~2.0 Active ingredient (GK2, etc.) Amount of active ingredient Active ingredients (plant extracts, etc.) 0.0001~5.0 Purified water (enough to make the total volume 100 parts)

[0036] Formulation example 9. Powder foundation [Ingredients] Part Iron oxide 0.1~5.0 Silica 0.1~20.0 Titanium oxide 0.1~20.0 Jojoba oil 0.1~10.0 Lactobacillus 0.0001~0.1 Plant extract (Melissa extract or Aloe vera extract) 0.0001~2.0 Active ingredient (GK2, etc.) Amount of active ingredient Active ingredients (plant extracts, etc.) 0.0001~5.0 Mica Amount that makes the total amount 100 parts

[0037] Prescription example 10. Body shampoo [Ingredients] Part Sodium lauroylmethyl-β-alanine solution 0.1~40.0 Coconut oil fatty acid diethanolamide 0.1~10.0 Phenoxyethanol 0.01~1.0 1,2-Hexanediol 1.5~3 Lactobacillus 0.0001~0.1 Plant extract (Melissa extract or Aloe vera extract) 0.0001~2.0 Active ingredient (GK2, etc.) Amount of active ingredient Active ingredients (plant extracts, etc.) 0.0001~5.0 Purified water (enough to make the total volume 100 parts)

[0038] Prescription example 11. Hair shampoo [Ingredients] Part Sodium N-cocoyl methyl taurate 0.1~40.0 Coconut oil fatty acid amidopropyl betaine solution 0.1~20.0 Coconut oil fatty acid diethanolamide 0.01~10.0 Phenoxyethanol 0.01~1.0 Citric acid 0.1~1.0 O-[2-hydroxy-3-(trimethylammonio)propyl]hydroxyethylcellulose chloride 0.1~2.0 1,2-Hexanediol 1.5~3 Lactobacillus 0.0001~0.1 Plant extract (Melissa extract or Aloe vera extract) 0.0001~2.0 Active ingredient (GK2, etc.) Amount of active ingredient Active ingredients (plant extracts, etc.) 0.0001~5.0 Purified water (enough to make the total volume 100 parts)

[0039] Prescription example 12. Hair conditioner [Ingredients] Part Distearyldimethylammonium chloride 0.1~10.0 Cetyl alcohol 0.1~10.0 Stearyl alcohol 0.1~10.0 Phenoxyethanol 0.01~1.0 1,3-butylene glycol 0.1~10.0 Lactobacillus 0.0001~0.1 Plant extract (Melissa extract or Aloe vera extract) 0.0001~2.0 Active ingredient (GK2, etc.) Amount of active ingredient Active ingredients (plant extracts, etc.) 0.0001~5.0 Purified water (enough to make the total volume 100 parts) [Industrial Applicability]

[0040] As explained above, it is possible to provide a new external skin preparation containing Lactobacillus casei subsp. casei strain 327 cells and a predetermined plant extract.

Claims

1. A skin external preparation comprising cells of Lactobacillus casei subsp. casei strain 327 deposited under accession number NITE BP-02707 and a Melissa extract.

2. A topical skin preparation comprising cells of Lactobacillus casei subsp. casei strain 327 deposited under accession number NITE BP-02707 and an aloe vera extract.

3. The agent according to claim 1 or 2, wherein the bacterial cells are killed bacterial cells.

4. The agent according to claim 1 or 2, which is for promoting the growth of Staphylococcus epidermidis.

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