WO2022186513A1 - Cosmetic composition for skin care through porphyrin production control and hyaluronidase control in skin microbiome - Google Patents

Abstract

The present disclosure provides a cosmetic composition which is configured to inhibit the growth of a skin microbiome harmful to skin health and to reduce the production of the detrimental factor porphyrin while having no influences on skin microbiomes not harmful to skin health, thus healthily rebalancing the skin microbiome. The present disclosure also provides a cosmetic composition capable of suppressing porphyrin production in strains that produce porphyrin. The present disclosure provides a skin care composition which is helpful for moisturizing the skin and for preventing skin aging by inhibiting the activity of hyaluronidase secreted from a skin microbiome to suppress the degradation of hyaluronic acid.

Description

Cosmetic composition for improving skin by controlling porphyrin production and hyaluronic acid degrading enzymes of skin flora

This application claims priority based on Korean Application No. 10-2021-0028329 filed on March 3, 2021, and all contents disclosed in the specification and drawings of the application are incorporated herein by reference.

The present invention relates to a cosmetic composition that can beneficially control (rebalance) the growth and metabolites of skin flora. The present invention also relates to a cosmetic composition capable of inhibiting the production of harmful substances on the skin, such as inhibiting the production of porphyrin in a porphyrin-producing strain, and inhibiting the hyaluronic acid degradation ability of a strain secreting a hyaluronic acid-degrading enzyme.

The skin contains millions of bacteria, fungi and viruses that make up the skin microbiome. The microbiome refers to the total number of microorganisms present in a specific environment. The genes of microorganisms that exist in the human body exist hundreds of times more than human genes, and they affect various functions in the human body.

Most of the human microbiota is known to exist in the intestine, but various microorganisms are also present in the skin. It is known that microorganisms present in the skin also play an essential role in various metabolisms such as pathogen invasion, immune system regulation, and natural product decomposition, similar to the microorganisms in the intestine, and can affect human health. The skin, the largest organ of the human body, is colonized by beneficial microorganisms and acts as a physical barrier to prevent the entry of pathogens. When the barrier is weakened or the balance between symbionts and pathogens is disrupted, skin diseases or systemic diseases can develop. Studying the composition of the human skin site microbiota is useful for elucidating the pathogenesis of common skin diseases or association with skin conditions.

If you use a fungicide or antibiotic with a strong bactericidal action that can eliminate all intrinsic microbial communities on the skin, you risk eliminating bacteria that have a beneficial effect on the skin as well, promoting the emergence and proliferation of bacteria resistant to that component. can be In addition, when the use of disinfectants or antibiotics is stopped, there is a risk of causing opportunistic infections and the like by preferentially growing undesirable bacteria on the skin because the existing skin flora balance is disrupted. If a substance that can selectively control the flora is used instead of broad-spectrum antibiotics, the proportion of bacteria having a beneficial effect on the skin can be improved while avoiding a wide range of side effects on the skin's intrinsic microbial community.

In other words, disruption of the balance between beneficial and harmful bacteria present on the skin can cause opportunistic infections. Therefore, maintaining the balance between microorganisms is an important part to maintain a healthy skin condition.

Cutibacterium acnes (C. acne) serves to inhibit the invasion or growth of pathogenic bacteria by decomposing sebum into glycerol and fatty acids to keep the skin slightly acidic. It is one of the major bacterial species that reside on the skin and plays an important role in maintaining the skin barrier. Therefore, it is undesirable to apply antibiotics with antibacterial or bactericidal action to the skin that can affect the entire C. acnes bacteria.

Recently, a phylogenetic analysis of acne bacteria has been performed, and the C. acne (Cutibacterium acne) strain, which is mainly possessed by healthy people rather than the acne patient group, and the C. acne strain, which is mainly present in acne patients without being present in healthy skin, has been identified. it became clear that In addition, C. acne has different characteristics of secreting harmful molecules such as protease, lipase, hemolysin and porphyrin depending on the strain, and through these, harmful reactions of the skin such as decomposing host tissues or inducing inflammatory reactions can cause

For example, porphyrin levels are observed to be higher in acne skin compared to healthy skin, and higher in acne lesions compared to non-lesion areas of acne patients. In particular, it is known that the strain of C. acnes related to acne produces a higher concentration of porphyrin. It is also known that porphyrin can be produced in the case of S. aureus, which is a problem in atopic dermatitis or wound infection.

Porphyrin not only induces oxidative stress in the skin and promotes the expression of inflammatory cytokines in keratinocytes, but also induces a carbonylation reaction, a type of protein oxidation, which can darken the skin tone and make the skin look unhealthy. Therefore, it is used as a measurement index in various types of skin measuring devices and needs to be controlled as one of the major harmful factors of skin microorganisms.

Therefore, the problem to be solved by the present invention is to inhibit the skin flora (for example, a specific C. acne strain) that has a relatively bad effect on the skin, such as causing acne, and has no significant effect on the skin ( For example, it is to provide a cosmetic composition capable of rebalancing the skin flora by having relatively little influence on the growth of a specific C. acnes strain).

Another object to be solved by the present invention is to provide a skin improvement composition that controls oxidative stress and inflammation of the skin by inhibiting the production of porphyrin secreted by microorganisms present in the skin.

Another object of the present invention is to provide a skin improvement composition capable of preventing hyaluronic acid decomposition by inhibiting hyaluronic acid degrading enzyme secreted by microorganisms.

In order to solve the above problems, the present disclosure provides a skin microbiome comprising at least one selected from the group consisting of thiamine or a cosmetically acceptable salt thereof and saccharide isomerate as an active ingredient. A cosmetic composition for conditioning is provided.

Another aspect of the present disclosure includes thiamine or a cosmetically acceptable salt thereof and any one or more selected from the group consisting of saccharide isomerate as an active ingredient, for controlling Cutibacterium acne in the skin microbiome A cosmetic composition is provided.

According to one aspect of the present disclosure, the cosmetic composition according to the present invention may be used for skin improvement through control of porphyrin production and/or control of hyaluronic acid degrading enzymes in skin flora. Accordingly, one aspect of the present disclosure provides a skin improvement composition that controls oxidative stress and inflammation of the skin by inhibiting the production of porphyrin secreted by microorganisms present in the skin to control harmful skin factors. Another aspect of the present disclosure provides a skin improvement composition capable of preventing hyaluronic acid decomposition by inhibiting hyaluronic acid degrading enzyme secreted by microorganisms.

Preferably, the cosmetic composition(s) according to the present disclosure includes thiamine or a cosmetically acceptable salt thereof as an active ingredient.

The present inventors tested various substances such as amino acids, amino acid derivatives, extracts, saccharides, vitamins, and fragrance ingredients, and completed the present invention by confirming that only the compounds of the present invention exhibit the desired effect in the present invention.

Fitz-Gibbon, S. etc., (2013). Propionibacterium acnes Strain Populations in the Human Skin Microbiome Associated with Acne. According to Journal of Investigative Dermatology, 133(9), 2152-2160, one of the skin flora, C. acnes, can be divided as follows, and RT4 and RT5 based on the ribotype are known as acne-inducing types, RT6 is known to be abundant in healthy skin.

Ribotype	RT4 (e.g., HL053PA1 strain)	RT6 (e.g., HL110PA3 strain)
cause acne	has exist	doesn't exist
incubation rate	Slow (6 days)	Medium (4 days)
porphyrin production	High (500nM)	Low (about 30 nM)
HA resolution	lowness	very high

The HL053PA1 strain and the like produce excessive amounts of porphyrin, which can cause inflammation and oxidative stress, which can cause the secretion of inflammatory cytokines in the skin to cause inflammation such as acne. Inhibits the growth of the induced type (eg, RT4 or RT5, type IA based on recA type) and has relatively no effect on the growth of the acne-inducing type (eg, RT6), so the dominant species microbiome ecology can be maintained.

In addition, the cosmetic composition of the present disclosure inhibits the porphyrin production of the skin flora, particularly the porphyrin production of the acne-causing type C. acnes strain or the porphyrin production of the strain such as S. aureus, thereby inhibiting the occurrence of acne, or the skin dermis is dense. It is very helpful in increasing the skin tone, improving the symptoms of dry skin, or improving skin tone.

According to a recent study, it was known that strains belonging to RT6 (or recA type standard type II) produce relatively few harmful factors such as porphyrin, but can produce an excess of hyaluronic acid degrading enzyme. It is also known that various bacteria such as S. aureus can produce hyaluronic acid degrading enzymes.

In this aspect, the cosmetic composition of the present disclosure is a cosmetic composition for improving skin elasticity through inhibition of hyaluronic acid decomposition of skin flora, in particular, inhibition of hyaluronic acid decomposition by non-acne type C. acne strain or S. aureus. may have beneficial effects. This inhibition of hyaluronic acid decomposition also improves the density of the dermis, thereby showing good effects in moisturizing and preventing aging.

"Cosmetically acceptable salts" in the present invention include salts of the active compounds prepared with relatively non-toxic acids. Acid addition salts may be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, neat or with a suitable inert solvent. Examples of cosmetically acceptable acid addition salts are acetic acid, propionic acid, isobutyric acid, oxalic acid, maleic, malonic, benzoic, succinic, suberic, fumaric ( fumaric), mandelic, phthalic, benzenesulfonic, p-tolylsulfonic, citric acid, tartaric acid, methanesulfonic, and the like. As well as salts derived from non-toxic organic acids, hydrogen chloride, hydrogen bromide, nitric acid, carbonic acid, monohydrogencarbonic, phosphoric, monohydrogenphosphate, dihydrogenphosphoric acid, sulfuric acid, monohydrogensulfuric acid, hydrogen iodide or phosphorous acid ( phosphorous acid) and its analogs. Also included are salts of amino acids such as arginate and analogues thereof and analogues of organic acids such as glucuronic or galactunoric acids and analogues thereof. Other examples of salts can be found in literature known in the art.

In the cosmetic composition according to the present invention, the total content of the active ingredients is preferably 0.00001 to 10% by weight based on the total weight of the cosmetic composition.

The active ingredient of the present invention may exist in unsolvated as well as solvated forms, including hydrates, ethanolates, and the like. The active ingredient of the present invention may exist in crystalline or amorphous form, and all such physical forms are included within the scope of the present invention.

The cosmetic composition according to the present invention includes solutions, external ointments, creams, foams, nourishing lotions, softening lotions, packs, soft water, emulsions, makeup bases, essences, soaps, liquid detergents, bathing agents, sunscreen creams, sun oils, suspensions, Emulsions, pastes, gels, lotions, powders, soaps, surfactant-containing cleansing, oils, powder foundations, emulsion foundations, wax foundations, patches and sprays may be prepared in a formulation selected from the group consisting of, but not limited to it is not

In addition, the cosmetic composition of the present invention may further include one or more cosmetically acceptable carriers to be formulated in general skin cosmetics, and conventional ingredients include, for example, oil, water, surfactant, humectant, lower alcohol, A thickener, a chelating agent, a colorant, a preservative, a fragrance, etc. may be appropriately mixed, but the present invention is not limited thereto.

The cosmetically acceptable carrier included in the cosmetic composition of the present invention varies depending on the formulation. When the formulation of the present invention is an ointment, paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide or Mixtures thereof may be used.

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder, or a mixture thereof may be used as a carrier component, and in particular, in the case of a spray, additional chloro propellants such as fluorohydrocarbons, propane/butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer, or emulsifier is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl benzoate, propylene glycol, 1,3 -Butylglycol oil, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, glycerol fatty esters, fatty acid esters of polyethylene glycol or sorbitan.

When the formulation of the present invention is a suspension, as a carrier component a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters; Crystalline cellulose, aluminum metahydroxide, bentonite, agar, or tracanth may be used.

When the formulation of the present invention is a soap, alkali metal salts of fatty acids, fatty acid hemiester salts, fatty acid protein hydrolysates, isethionate, lanolin derivatives, aliphatic alcohols, vegetable oils, glycerol, sugars, etc. may be used as carrier components. can

All ingredients described in the present invention, preferably, relevant laws and regulations of Korea, China, the United States, Europe, Japan, etc. (eg, regulations on cosmetic safety standards (Korea), cosmetic safety technical standards (China), Do not exceed the maximum use values stipulated in the Food Standards Code (Korea), Food Additives Code (Korea), Health Functional Food Code (Korea), and hygiene standards (China)). That is, preferably, the composition for cosmetics, food, or personal care according to the present invention contains the ingredients according to the present invention within the content limit allowed by the relevant laws and regulations of each country.

The composition provided in the present disclosure, particularly the cosmetic composition, has an effect of inhibiting the growth of major flora of the skin, for example, the growth of C. acne type frequently present in acne patients among C. acnes and the production of harmful factors (porphyrin). indicates. In addition, it can exhibit the effect of inhibiting the activity of hyaluronidase produced by this strain without affecting the growth of C. acne type, which is mainly present in non-inflammatory skin. In addition, it may exhibit an effect of inhibiting porphyrin production and/or hyaluronidase activity generated by bacteria present in the skin, such as S. aureus, in addition to the C. ance strain.

The following drawings attached to the present specification illustrate preferred embodiments of the present invention, and serve to further understand the technical idea of the present invention together with the above-described contents of the present invention, so the present invention is limited to the matters described in such drawings It should not be construed as being limited.

1 is a result of evaluating the effect of thiamine salt on the pathogenic strain Cutibacterium acnes.

Figure 2 is a result of evaluating the effect of the thiamine salt on a non-pathogenic strain of Cutibacterium acnes.

Figure 3 is a result of evaluating the effect of the thiamine salt on the production of porphyrin of the pathogenic and non-pathogenic strains of Cutibacterium acnes.

4 is a result of evaluating the effect of thiamine salt on porphyrin production of Staphylococcus aureus strain.

5 is a result of evaluating the effect of the thiamine salt on the hyaluronic acid decomposition ability of a non-pathogenic strain of Cutibacterium acnes.

6 is a result of evaluating the effect of the thiamine salt on the hyaluronic acid degradation ability of Staphylococcus aureus strain.

7 is a result of evaluating the effect of saccharide isomerate on the growth of pathogenic and non-pathogenic strains of Cutibacterium acnes and on the production of porphyrins of pathogenic strains of Cutibacterium acne. In FIG. 7, a is a result of evaluating the effect on the strain growth of HL053PA1, a C. acnes strain associated with acne, and b is a result of evaluating the effect on the growth of a strain of HL110PA3, a healthy skin-related C. acnes strain. , c is the result of evaluating the effect on porphyrin production of HL053PA1, a C. acnes strain associated with acne skin, and d is the result of evaluating the effect on the degradation rate of hyaluronic acid of HL110PA3, a healthy skin-related C. acne strain.

Hereinafter, examples and the like will be described in detail to help the understanding of the present invention. However, the embodiments according to the present invention may be modified in various other forms, and the scope of the present invention should not be construed as being limited to the following examples. The embodiments of the present invention are provided to more completely explain the present invention to those of ordinary skill in the art to which the present invention pertains.

For reference, materials and methods other than those specifically mentioned among the following experimental methods are described in the paper "Vitamin B12 modulates the transcriptome of the skin microbiota in acne pathogenesis" (Sci Transl Med. 2015 Jun 24;7(293):293ra103. doi: 10.1126/scitranslmed.aab2009.) or "The Skin Bacterium Propionibacterium acnes Employs Two Variants of Hyaluronate Lyase with Distinct Properties" (Microorganisms. 2017 Sep 12;5(3):57. doi: 10.3390/microorganisms5030057.) .

1. Acne-associated C. acne strain growth inhibition (Fig. 1)

Experimental method : An experiment was performed to determine the effect of Thiamine HCl on the proliferation of acne-related C. acnes.

The HL053PA1 strain, which is a Cutibacterium acne type, which is mainly present in acne patients, was cultured and activated in BHI broth, and then re-inoculated at the same turbidity. At this time, thiamine HCl was treated at a concentration of 1 w/v% and absorbance was measured at 595 nm after incubation for 7 days to confirm the effect on strain growth. The results are shown in FIG. 1 .

Result : Thiamine HCl treatment reduced the growth by about 70% compared to the untreated group, confirming that the growth of C. acne type, which mainly emits harmful factors, was inhibited.

2. Normal skin-associated C. acne strain growth (Fig. 2)

Experimental method : An experiment was performed to confirm the effect of Thiamine HCl on the proliferation of C. acne type, which is mainly present only in normal skin without inflammation such as acne.

The HL110PA3 strain, which is a C. ance type mainly present only in normal skin, was cultured and activated in BHI liquid medium, and then re-inoculated at the same turbidity. At this time, thiamine HCl was treated at a concentration of 1%, and absorbance was measured at 595 nm after 7 days of incubation to confirm the effect on strain growth. The results are shown in FIG. 2 .

Results : Thiamine HCl treatment significantly reduced the growth of acne-related C. acne type, but had no significant effect on the growth of C. acne type, a normal skin type.

3. Inhibitory effect of porphyrin production (Fig. 3)

Experimental method: An experiment was conducted to investigate the effect on the production of porphyrin, which is known to induce oxidative stress and is involved in inflammation such as acne in the skin.

C. acnes HL053PA1 and HL110PA3 strains were respectively cultured and activated in BHI broth, and then re-inoculated at the same turbidity. At this time, thiamine HCl was treated at a concentration of 1%, and the strain culture was recovered after 7 days of incubation. 400 μl of the bacterial culture was extracted by adding ethyl acetate and acetic acid (4: 1, vol/vol). The extract was centrifuged to recover the upper layer containing porphyrin through layer separation. The recovered supernatant was dissolved in 1.5 M HCl, and the layers were separated by centrifugation to recover the lower layer. The recovered soluble phase was measured for fluorescence at 405 nm/620 nm. The results are shown in FIG. 3 .

RESULTS: The acne-associated C. acnes strain produced a greater amount of porphyrin than the C. acnes strain present in normal skin. Thiamine HCl treatment significantly reduced the amount of porphyrin present in the culture of acne-associated strain HL053PA1.

4. Inhibitory effect of porphyrin production in other porphyrin-producing strains (Fig. 4)

S. aureus strains were re-inoculated at 1/100 concentration activated by culturing in BHI broth. The strain culture was recovered after treatment with thiamine HCl at a concentration of 1 w/v% and culturing for 24 hours. 400 μl of the bacterial culture was extracted by adding ethyl acetate and acetic acid (4: 1, vol/vol). The extract was centrifuged to recover the upper layer containing porphyrin through layer separation. The recovered supernatant was dissolved in 1.5 M HCl, and the layers were separated by centrifugation to recover the lower layer. The recovered soluble phase was measured for fluorescence at 405 nm/620 nm. The results are shown in FIG. 4 .

Results: Thiamine HCl treatment significantly reduced the amount of porphyrin secreted by S. aureus.

5. Hyaluronic acid degradation inhibitory effect (Fig. 5)

Experimental method: An experiment was performed to determine whether the activity of the hyaluronic acid degrading enzyme secreted by HL110PA3, a general skin-associated C. acnes strain, is inhibited.

The HL110PA3 strain was activated by culturing in BHI broth and then re-inoculated to the same turbidity. At this time, thiamine HCl was treated at a concentration of 1%, and the culture medium was recovered after 7 days of incubation. In order to measure the activity of the hyaluronic acid degrading enzyme secreted by C. acne, the hyaluronic acid and the culture medium were mixed and reacted at 37° C. for 20 minutes. After the reaction was completed, 1% BSA sodium acetate buffer was added to aggregate the remaining hyaluronic acid and BSA, and the degree of decomposition of hyaluronic acid was confirmed by taking absorbance at 540 nm. The results are shown in FIG. 5 .

Result: HL110PA3, a non-pathogenic strain of C. acnes, is generally classified as a healthy skin type C. acne because it is isolated from healthy skin without inflammatory lesions. can do. While about 85% of hyaluronic acid was decomposed for 20 minutes when treated with untreated culture medium, about 60% of hyaluronic acid was decomposed in the culture medium of the experimental group treated with Thiamine HCl, indicating that it can inhibit the degradation of hyaluronic acid by 30% compared to the control group. Confirmed.

6. Efficacy of inhibiting hyaluronic acid degradation in other strains secreting hyaluronic acid degrading enzyme (Fig. 6)

Experimental method: An experiment was performed to determine whether the activity of S. aureus hyaluronic acid degrading enzyme was inhibited.

S. aureus strains were re-inoculated at 1/100 concentration activated by culturing in BHI broth. After treatment with thiamine HCl at a concentration of 1% and cultured for 24 hours, the culture medium was recovered. To measure the activity of the hyaluronic acid degrading enzyme secreted by S. aureus, the hyaluronic acid and the culture medium were mixed and reacted at 37° C. for 90 minutes. In order to measure the hyaluronic acid remaining without degradation, the remaining hyaluronic acid was quantified using a hyaluronan ELISA kit.

Results: S. aureus, which is found frequently in atopic dermatitis lesions, etc., can degrade hyaluronic acid. It was confirmed that more than 50% of hyaluronic acid was decomposed when treated with untreated culture, whereas less than 10% of hyaluronic acid was decomposed in the culture of the experimental group treated with Thiamine HCl, thereby significantly inhibiting the decomposition of hyaluronic acid.

7. Evaluation result of saccharide isomerate (Fig. 7)

In the same manner as the thiamine salt, the effect of saccharide isomerate on the growth of pathogenic and non-pathogenic strains of C. acnes acne was evaluated. The results are shown in FIG. 7 . As saccharide isomerate, PENTAVIIN® product was used.

As shown in FIG. 7 , while the saccharide isomerate also inhibited the growth of acne-causing harmful C. acnes bacteria, the growth inhibition of beneficial C. acnes bacteria was relatively small, confirming that it had a strain rebalancing effect. In addition, it was confirmed that the inhibitory effect of porphyrin production and hyaluronic acid degrading enzyme was exhibited.

Claims (7)

1. A cosmetic composition for adjusting skin microbiome, comprising as an active ingredient at least one selected from the group consisting of thiamine or a cosmetically acceptable salt thereof and saccharide isomerate.
2. The cosmetic composition according to claim 1, wherein the cosmetic composition is for adjusting Cutibacterium acnes in the skin microbiome.
3. The cosmetic composition according to claim 2, wherein the composition inhibits the growth of the Cutibacterium acne HL053PA1 strain more than the Cutibacterium acne HL110PA3 strain.
4. A cosmetic composition for inhibiting the production of porphyrin secreted by microorganisms, comprising as an active ingredient at least one selected from the group consisting of thiamine or a cosmetically acceptable salt thereof and saccharide isomerate.
5. The method according to claim 3, wherein the cosmetic composition is used for inhibiting skin oxidative stress, improving skin inflammation, inhibiting acne induction, increasing dermal density, improving skin dryness symptoms, or improving skin tone. , cosmetic composition.
6. A cosmetic composition for inhibiting hyaluronic acid degrading enzyme secreted by microorganisms, comprising as an active ingredient any one or more selected from the group consisting of thiamine or a cosmetically acceptable salt thereof and saccharide isomerate.
7. The cosmetic composition according to claim 6, wherein the composition is used to improve skin elasticity, dryness, or aging symptoms by inhibiting hyaluronic acid decomposition.

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