JP5972732B2 - Antibacterial agent and external preparation for skin - Google Patents

Description

  The present invention relates to an antibacterial agent and a skin external preparation.

In human skin, there are various microorganisms called skin resident bacteria. Although skin resident bacteria are useful in terms of preventing the invasion of pathogenic bacteria and maintaining the barrier function of the skin, some have been pointed out to be involved in skin diseases such as dermatitis.
Propionibacterium acnes , one of Gram-positive bacteria, plays a role in suppressing the invasion and propagation of pathogenic bacteria by breaking down sebum into glycerin and fatty acids to keep the skin weakly acidic. However, if sebum continues to clog pores, acne bacteria grow using sebum as a nutrient source, which causes inflammation such as acne.
Similarly, staphylococci (Bacteria of the genus Staphylococcus ), which is one of Gram-positive bacteria, are present in human skin, nasal cavity, etc., and are usually harmless bacteria. However, a highly toxic substance such as Staphylococcus aureus is known to cause suppuration and jumping of the wound. Staphylococcus aureus may enter the body from an inflamed wound and cause meningitis, pneumonia, peritonitis, sepsis and the like. In addition, it has been pointed out that it is a cause of worsening atopic dermatitis (see, for example, Non-Patent Documents 1 to 5).
Malassezia fungus (genus Malassezia ), one of the fungi, is ubiquitous on the surface of animals and humans and is said to cause folding screen, malassezia folliculitis, seborrheic dermatitis, and atopic dermatitis. ing. Malassezia fungi present in the scalp produce lipase to break down and consume the sebum of the scalp, and the fatty acids produced when this lipase breaks down sebum is said to cause scalp inflammation. . Therefore, it is considered that when a Malassezia fungus grows, lipase is excessively produced and the amount of fatty acid increases, thereby causing inflammation of the scalp, resulting in dandruff (for example, non-patent documents 6 to 8). reference).

  In order to prevent and improve skin inflammation, it is important to effectively control the growth of pathogenic bacteria, and the development of new antibacterial agents and external skin preparations useful for maintaining and improving skin health is hoped for. It is rare. For example, Patent Document 1 and Non-Patent Document 9 describe a skin external preparation containing a compound having anti-acne fungal activity or anti-S. Aureus activity.

JP 2010-65014 A

Journal of Allergy and Clinical Immunology, 2000, Vol. 105 p. 814. Journal of the Japanese Dermatological Association, 2000, Vol. 110, p. 19. British Journal of Dermatology, 1974, Vol. 90, p. 525. Skin (New: Skin Science), 1988, Vol. 40, p. 9. Skin Science, 2008, Vol. 10, p. 24. Journal of Clinical Microbiology, 2002, Vol. 40, p. 3350. Japanese Journal of Medical Mycology, 2007, Vol. 48, p. 179. Microbiology, 2011, Vol. 157, p. 3492. Phytochemistry, 2010, Vol. 71, p. 104.

  An object of the present invention is to provide an antibacterial agent and an external preparation for skin using a plant extract, particularly an antibacterial agent and an external preparation for skin that exhibit excellent antibacterial activity against causative bacteria of skin inflammation.

  In view of the above problems, the present inventors have conducted intensive studies. As a result, it was found that a plant extract of Malvata shows that it has excellent antibacterial activity against causative bacteria such as skin inflammation. The present invention has been completed based on this finding.

The present invention relates to an antibacterial agent containing an extract of Ainsliaea fragrans Champ as an active ingredient.
The present invention also relates to an external preparation for skin containing an extract of Ainsliaea fragrans Champ as an active ingredient.
The present invention also relates to an acne prevention / amelioration agent containing an extract of Ainsliaea fragrans Champ as an active ingredient.
The present invention also relates to a preventive / ameliorating agent for atopic dermatitis containing an extract of Ainsliaea fragrans Champ as an active ingredient.
The present invention also relates to a dandruff preventive / ameliorating agent containing an extract of Ainsliaea fragrans Champ as an active ingredient.

  The antibacterial agent and the external preparation for skin of the present invention exhibit excellent antibacterial activity, particularly excellent antibacterial activity against bacteria causing skin inflammation. In particular, the antibacterial agent and the external preparation for skin of the present invention exhibit excellent antibacterial activity against bacteria that cause skin inflammation causing acne, atopy, dandruff and the like.

The antibacterial agent and the external preparation for skin of the present invention contain an extract of Ainsliaea fragrans Champ as an active ingredient.
Malvata is a plant belonging to the genus Compositae.
For the extraction, any part (eg, whole grass, root, stem, leaf, flower, etc.) of the malt mushroom can be used, and a plurality of each part may be used in combination. Especially, it is preferable to use the whole plant or the site | part containing a root, and it is more preferable to use a root. In addition, it is also preferable to use a crude drug (Kinbetoji) obtained from Malvata shows as a base plant. It is known that this herbal medicine Kanabe Sakai exhibits clean fever and detoxification.
For the extraction, the above plant may be used as it is, and it is also preferable to use a plant that has been subjected to a treatment such as drying, shredding, or pulverization in order to increase extraction efficiency.

  There is no particular limitation on the extraction method, and the extraction can be performed by a normal plant extraction method. Specifically, dried products obtained by drying the above plants, squeezed juice obtained by squeezing and extracting the pulverized products, steam distillates, crude extracts using various extraction solvents, distribution of crude extracts, column chromatography, etc. Extract fractions obtained by purification by various chromatographic methods can be used as the extract in the present invention. Moreover, you may use what combined these extract, steam distillate, pressing material, etc. individually or in combination of 2 or more types as an extract.

In the present invention, it is more preferable to use an extract obtained by using an extraction solvent from a dried product obtained by drying the plant or a pulverized product thereof.
As the extraction solvent, either a polar solvent or a nonpolar solvent can be used, and these can also be mixed and used. For example, water; alcohols such as methanol, ethanol, propanol and butanol; polyhydric alcohols such as ethylene glycol, propylene glycol and butylene glycol; ketones such as acetone and methyl ethyl ketone; esters such as methyl acetate and ethyl acetate; tetrahydrofuran Linear and cyclic ethers such as diethyl ether; polyethers such as polyethylene glycol; halogenated hydrocarbons such as dichloromethane, chloroform and carbon tetrachloride; hydrocarbons such as hexane, cyclohexane and petroleum ether; benzene and toluene Aromatic hydrocarbons such as; pyridines; supercritical carbon dioxide; fats and oils, waxes, and other oils. Or the mixture which combined 2 or more types of the said solvent can be used as an extraction solvent. Of these, water, alcohols, alcohol aqueous solutions, propylene glycol, butylene glycol are preferably used, water, alcohols, alcohol aqueous solutions are more preferably used, alcohol aqueous solutions are more preferably used, and ethanol aqueous solutions are used. Is particularly preferred. The concentration of the ethanol aqueous solution is preferably 0.1 to 100 (v / v)%, more preferably 20 to 100 (v / v)%, and still more preferably 50 to 100%.
It is preferable to immerse or heat-reflux the plant as an extraction raw material in these solvents to obtain an extract.

  The conditions for solvent extraction differ depending on the solvent to be used and are not particularly limited. For example, when water, alcohols, aqueous alcohol solution, propylene glycol, or butylene glycol is used as the solvent, 1 part by weight or more and 50 parts by weight with respect to 1 part by weight of the plant. It is preferable to use a solvent having a volume part or less. The extraction temperature is preferably 3 ° C or higher, more preferably 20 ° C or higher. And preferably it is 100 degrees C or less, More preferably, it is 80 degrees C or less. The extraction time is preferably 1 hour or longer, more preferably 1 day or longer. And it is preferably about several weeks or less, more preferably about 30 days or less. Moreover, in order to raise extraction efficiency, you may stir together or may perform a homogenization process in a solvent.

  The extract obtained by the above-described method may be used as it is, and the active extract by an appropriate separation means such as gel filtration, chromatography, precision distillation or the like may be used. A high fraction can also be fractionated and used. Moreover, what diluted or concentrated the said extract and fraction may be used, and after drying by freeze-drying etc., it may be made into the powder or the paste form. Examples of the extract of Malvata commune used in the present invention include various extracts obtained by the above-described method, diluted solutions thereof, concentrated solutions thereof, purified products thereof, dried powders thereof and the like.

  As shown in the below-mentioned Examples, the plant extract obtained from Malvata shows an excellent antibacterial action and can be used as an antibacterial agent. In the present invention, the term “antibacterial” includes both the concepts of inhibiting the growth of bacteria and killing the bacteria, and the bacteria include both bacteria and fungi. Further, the use may be a therapeutic use (ie medical practice) or a non-therapeutic use (non-medical practice). The subject of the use can be a human, a non-human animal, or a specimen derived therefrom.

In particular, the extract of Malvata shows an excellent antibacterial activity against bacteria that cause inflammation of the skin. Examples of bacteria that cause skin inflammation include acne-causing bacteria, atopic dermatitis-causing bacteria, and dandruff-causing bacteria. More specifically, acne bacteria as acne-causing bacteria (Propionibacterium acnes) is, Staphylococcus aureus (Staphylococcus aureus) or Malassezia (Malassezia) spp fungi as atopic dermatitis caused bacteria, Marasechi genus as dandruff-causing bacteria A fungus is mentioned. The antibacterial agent of the present invention can be suitably used as an antibacterial agent against these bacteria.

  The antibacterial agent of the present invention can be applied for both therapeutic use (medical use) and non-therapeutic use (non-medical use), and the use form thereof is human and animal skin, nails, mucous membrane, hair, etc. All forms that can be applied to are included. Specific examples include use as pharmaceuticals, quasi-drugs, cosmetics, etc., in the case of cosmetics, quasi-drugs, and pharmaceuticals, in the form of external skin preparations, and in the case of pharmaceuticals, in the dosage form can do.

The antibacterial agent of the present invention may contain only the above-mentioned extract of the maltflower extract, which is an active ingredient, or may contain other ingredients as long as the effect is not affected. Good.
Components other than the active ingredient can be appropriately selected according to the form and use of the agent. For example, an appropriate liquid or solid excipient or extender such as titanium oxide, calcium carbonate, distilled water, lactose, starch or the like may be added so as to facilitate forming as an agent. In addition, other medicinal components (whitening agents, moisturizers, etc.), antioxidants, ultraviolet absorbers, surfactants, thickeners, coloring materials and the like may be added.

  When other ingredients are blended with the agent, the content of the extract of the extract is not particularly limited, but the above extract is contained in the agent in a solid content concentration (in terms of solid content) of 0.0001% by mass or more. It is preferable that the content is 0.005% by mass or more, and it is particularly preferable that the content is 0.01% by mass or more. Moreover, it is preferable that 20 mass% or less of the said extract is contained by solid content concentration in an agent, and it is more preferable that 10 mass% or less is contained.

When the antibacterial agent of the present invention is used in the form of a skin external preparation, the “skin external preparation” includes those applied to the skin as skin cosmetics, external pharmaceuticals, quasi-drugs, etc. The dosage form may take a wide variety of forms such as an aqueous solution system, a solubilization system, an emulsification system, a powder system, a gel system, an ointment system, a cream, a water-oil two-layer system, and a water-oil-powder three-layer system. More specifically, facial cleanser, lotion, lotion, milky lotion, cream, gel, essence (beauty serum), pack, mask, massage agent, foundation, lipstick, bath preparation, shampoo, hair conditioner, hair tonic, ointment, Examples include pastes, sheet-like products (absorbent articles such as sanitary products, wiping wipes, wet tissues, etc.).
In addition, in making these dosage forms, various oil agents, surfactants, gelling agents, preservatives, antioxidants, solvents, alcohol, water, chelating agents, thickeners, ultraviolet absorbers, emulsion stabilizers, pH A regulator, a pigment, a fragrance, and the like can be appropriately blended.

When the antibacterial agent of the present invention is used as a pharmaceutical in a form to be administered to humans or animals, for example, oral solid preparations such as tablets, coated tablets, granules, powders, capsules, oral administration liquids, syrups, etc. It can be in the form of parenteral preparations such as liquid preparations, injections, external preparations, suppositories, and transdermal absorption agents.
In preparing these preparations, excipients, binders, extenders, disintegrants, surfactants, lubricants, dispersants, buffers, preservatives, flavoring agents, fragrances, coating agents, carriers, Diluents and the like can be appropriately blended.

When the antibacterial agent of the present invention is used in the form of a skin external preparation, the amount of the skin external preparation used varies depending on the content of the active ingredient in the preparation, but is 0.0001 mg or more and 20 mg or less per 1 cm ^{2 of} the skin surface. It is preferable to do.
In addition, when the antibacterial agent of the present invention is used in the form of a pharmaceutical or the like, the amount used varies depending on the condition of the subject person, weight, sex, age, etc., but usually per adult (weight 60 kg). It is preferable to use 0.001 mg or more and 1000 mg or less of the extract of Malvata per day in terms of dry solid content.

  The antibacterial agent and the external preparation for skin of the present invention comprise an extract of maltose ginger extract as an active ingredient, and has an excellent antibacterial action against causative bacteria of dermatitis, particularly acne-causing bacteria, atopic dermatitis-causing bacteria, and dandruff-causing bacteria. Indicates. Due to this antibacterial action, the antibacterial agent and skin external preparation of the present invention prevent, alleviate and improve various symptoms caused by skin inflammation caused by these causative bacteria, such as acne, suppuration, atopy and dandruff. Can do. From this viewpoint, the antibacterial agent and the external preparation for skin of the present invention prevent the onset of atopic dermatitis as an acne preventive agent or an improving agent for preventing the development of acne or alleviating or improving the symptoms of acne, or As an atopic dermatitis preventive or ameliorating agent for alleviating or improving the symptoms of atopic dermatitis, to be used as a dandruff preventing or improving agent for preventing the occurrence of dandruff or for reducing or improving dandruff symptoms Can do.

  The present invention further discloses the following antibacterial agents, external preparations for skin, methods and uses for the embodiments described above.

<1> An antibacterial agent containing an extract of Ainsliaea fragrans Champ as an active ingredient.
<2> The antibacterial agent according to <1>, which is an antibacterial agent against acne-causing bacteria.
<3> The antibacterial agent according to <1> or <2>, which is an antibacterial agent against Propionibacterium acnes .
<4> The antibacterial agent according to <1>, which is an antibacterial agent against atopic dermatitis-causing bacteria.
<5> The antibacterial agent according to <1> or <4>, which is an antibacterial agent against Staphylococcus aureus .
<6> The antibacterial agent according to <1>, which is an antibacterial agent against dandruff-causing bacteria.
<7> The antibacterial agent according to <1> or <6>, which is an antibacterial agent against a fungus of the genus Malassezia .
<8> The antibacterial agent according to <1>, wherein the extract is extracted using an aqueous alcohol solution as a solvent.
<9> An antibacterial method (excluding medical practice), characterized by applying cosmetics containing an extract of Ainsliaea fragrans Champ to the skin.

<10> A method of using an extract of Ainsliaea fragrans Champ as an antibacterial agent.
<11> Use of an extract of Ainsliaea fragrans Champ as an antibacterial agent.
<12> Non-pharmaceutical use of an extract of Ainsliaea fragrans Champ for antibacterial purposes.
<13> Use of an extract of Ainsliaea fragrans Champ for the production of an antibacterial agent.
<14> An extract of Ainsliaea fragrans Champ used for antibacterial purposes.
<15> An antibacterial method comprising administering or applying to the skin an extract of Ainsliaea fragrans Champ.

<16> An external preparation for skin containing an extract of Ainsliaea fragrans Champ as an active ingredient.
<17> The external preparation for skin according to <16>, wherein the extract is extracted using an aqueous alcohol solution as a solvent.

<18> An acne prevention / amelioration agent comprising an extract of Ainsliaea fragrans Champ as an active ingredient.
<19> An atopic dermatitis preventive / ameliorating agent containing an extract of Ainsliaea fragrans Champ as an active ingredient.
<20> A dandruff preventive / improving agent comprising an extract of Ainsliaea fragrans Champ as an active ingredient.

  EXAMPLES Hereinafter, although this invention is demonstrated further in detail based on an Example, this invention is not limited to this.

Production Example Preparation of Marbateishosou Extract Herbal medicine based on Marbateishoso, Kinpentoji (Shinwa product, Marbateishoso whole plant) 160g, shredded 95g It was immersed in 1600 mL of a volume% ethanol aqueous solution for 7 days at room temperature. After filtering this immersion liquid, the solvent was removed to obtain 2.89 g of a solid content. This solid content was diluted with a 95% aqueous ethanol solution so that the evaporation residue was 1.0 w / v%, to prepare an extract of Marbata perforatum.

Test Example 1 Antibacterial Test against Acne Bacteria Antibacterial activity against acne bacteria was evaluated by using the extract of Malvata shows. Antibacterial activity was evaluated using a commercially available kit (trade name: BacTiter-Glo ^™ Microbial Cell Viability Assay, manufactured by Promega) for quantifying the amount of ATP using luciferase.
A 96-well plate containing 10 μL of a culture solution of Acne JCM6425 strain anaerobically cultured at 37 ° C. for 3 days using a modified GAM medium (manufactured by Nissui Pharmaceutical Co., Ltd.), 80 μL of the modified GAM medium, and 10 μL of the extract And anaerobically cultured at 37 ° C. for 24 hours. The concentration of the extract of Malvata shows in this culture solution is 0.1 w / v%. After incubation, 100 μL of BacTiter-Glo ^™ reagent was added to the wells, and the fluorescence intensity (excitation light: 488 nm) at 530 nm was quantified using a 1420 Multilabel Counter (manufactured by Wallac).
A similar test was conducted using a system in which 95% ethanol was mixed in place of the extract of Marbateisoba as a negative control. In addition, a similar test was performed using a mixture of isopropylmethylphenol (IPMP) to a final concentration of 1 mM as a positive control in place of the extract of Malvata. Isopropylmethylphenol (IPMP) is a compound known to exhibit a broad antibacterial spectrum, and is generally used as an active ingredient of various bactericidal disinfectants and acne remedies.
The results are shown in Table 1.

  As shown in Table 1, the fluorescence intensity was much lower in the system containing the extract of the malt mushroom extract compared to the negative control, and the growth of acne bacteria was remarkably suppressed. This growth inhibitory effect was superior to the positive control. From the above results, it was found that the extract of Malvata shows an excellent antibacterial activity against acne bacteria.

Test Example 2 Antibacterial test against Staphylococcus aureus The antibacterial activity against Staphylococcus aureus was evaluated using the extract of Malvata serrata prepared in the above production example. Antibacterial activity was evaluated using a commercially available kit (trade name: BacTiter-Glo ^™ Microbial Cell Viability Assay, manufactured by Progega) that quantifies ATP using luciferase.
10 μL of the culture solution of S. aureus IFO13276 strain aerobically cultured at 37 ° C. for 1 day using SCD medium (manufactured by Daigo), 80 μL of SCD medium, and 10 μL of Marbata serrata extract in a well of a 96-well plate The mixture was mixed and aerobically cultured at 37 ° C. for 24 hours. The concentration of the extract of Malvata shows in this culture solution is 0.1 w / v%. After incubation, 100 μL of BacTiter-Glo ^™ reagent was added to the wells, and the fluorescence intensity at 530 nm was quantified using a 1420 Multilabel Counter (manufactured by Wallac).
A similar test was conducted using a system in which 95% ethanol was mixed in place of the extract of Marbateisoba as a negative control. In addition, a similar test was performed using a mixture of isopropylmethylphenol (IPMP) to a final concentration of 1 mM as a positive control in place of the extract of Malvata. Isopropylmethylphenol (IPMP) is a compound known to exhibit a broad antibacterial spectrum, and is known to exhibit antibacterial effects against Staphylococcus aureus. Moreover, it is generally used as an active ingredient of a disinfectant.
The results are shown in Table 2 below.

  As shown in Table 2, the fluorescence intensity was much lower in the system containing the extract of the malt mushroom extract compared to the negative control, and the growth of Staphylococcus aureus was remarkably suppressed. This growth inhibitory effect was much superior to the positive control. From the above results, it was found that the extract of Malvata shows an excellent antibacterial activity against Staphylococcus aureus.

Test Example 3 Antibacterial test against Malassezia fungus Antibacterial activity against Malassezia fungus was evaluated using the extract of Malvatia spp. Prepared in the above production example. Antibacterial activity was evaluated using a commercially available kit (trade name: BacTiter-Glo ^™ Microbial Cell Viability Assay, manufactured by Promega) for quantifying the amount of ATP using luciferase.
Using a modified LNA medium (see Japanese Journal of Medical Mycology 2007, Vol. 48, p. 179), the B. malassezia obtained by stationary culture at 32 ° C. for 7 days: Malassezia globosa CBS7874 strain in a 96-well plate To each well, 90 μL of a 0.5% Tween 60 (manufactured by Kanto Chemical Co., Ltd.) aqueous solution and 10 μL of the extract of Mulberry spruce were added and mixed, followed by shaking culture at 32 ° C. for 24 hours. The concentration of the extract of Malvata shows in this culture solution is 0.1 w / v%. After incubation, 100 μL of BacTiter-Glo ^™ reagent was added to the wells, and the fluorescence intensity at 530 nm was quantified using a 1420 Multilabel Counter (manufactured by Wallac).
A similar test was conducted using a system in which 95% ethanol was mixed in place of the extract of Marbateisoba as a negative control. In addition, a similar test was performed using a mixture of miconazole nitrate to a final concentration of 0.4 mM as a positive control in place of the extract of Malvata. Miconazole nitrate is a compound known to have an effect of suppressing the occurrence of dandruff, and is used as an active ingredient of anti-dandruff shampoo.
The results are shown in Table 3 below.

  As shown in Table 3, the fluorescence intensity was much lower in the system containing the extract of Malvata serrata than in the negative control, and the growth of Malassezia fungi was remarkably suppressed. This growth inhibitory effect was much superior to the positive control. From the above results, it was found that the extract of Malvata shows that it has an excellent antibacterial activity against Malassezia fungi.

(Prescription example)
As an example of the usage form of the anti-acne fungal agent of the present invention, lotions, creams and ointments having the compositions shown below can be prepared by conventional methods.

1. Preparation of lotion (composition) (blending amount: mass%)
Marbateishoso extract 0.1
Glycerin 10
1,3-Butylene glycol 6
Citric acid 0.1
Sodium citrate 0.3
Ethyl alcohol 8
Polyoxyethylene hydrogenated castor oil (60EO) 0.5
Perfume
Water remaining
Total 100

2. Preparation of cream (composition) (blending amount: mass%)
Marbateishoso extract 1
Monostearic glyceride 2
Polyoxyethylene sorbitan monolaurate (20EO) 2
Cetanol 7
Squalene 40
Propylene glycol 10
Liquid paraffin 10
Perfume
Total remaining water 100

3. Preparation of ointment (Composition) (Amount: Mass%)
Marbateishoso extract 2
Sara beeswax 11
Liquid paraffin 22
Lanolin 10
Polysorbate 80 2
Olive oil 15
Preservative appropriate amount
Total remaining water 100

Claims (5)

An antibacterial agent against Propionibacterium acnes, containing an extract of Ainsliaea fragrans Champ as an active ingredient. The antibacterial agent of Claim 1 whose said acne microbe (Propionibacterium acnes) is an acne causative microbe . An antibacterial agent against dandruff-causing bacteria , containing an extract of Ainsliaea fragrans Champ as an active ingredient . The antibacterial agent according to claim 3 , which is an antibacterial agent against Malassezia fungus. An antidandruff agent / improvement agent containing an extract of Ainsliaea fragrans Champ as an active ingredient.