Composition containing HMO

Abstract

The invention relates to application of fucosyllactose in preparation of a composition for improving HaCaT cell AQP3 expression, and belongs to the technical field of HMO (s) application and the technical field of beauty cosmetics. The invention provides an application of fucosyllactose in preparation of beauty cosmetics for improving AQP3 expression of HaCaT cells. The fucosyllactose comprises, but is not limited to, 2 '-fucosyllactose (2'-fucosyllactose, 2 '-FL), 3-fucosyllactose (3-fucosyllactose, 3-FL), difucosyllactose (Difucosyllactose, DFL), lactose-N-fucopentose I (Lacto-N-fucopentose I, LNFP I), and lactose-N-difucohexose I (Lacto-N-difucohexose I, LNDFH I), and the fucosyllactose comprises, but is not limited to, 2'-fucosyllactose (2 '-fucosyllactose, 2'-FL), 3-fucosyllactose ( The invention further provides a beauty makeup product with a moisturizing function. The beauty makeup product has an excellent moisturizing effect, is safe and reliable, has no irritation to skin, and does not generate anaphylactic reaction.

Description

Technical Field

[0001] The invention relates to application of fucosyllactose in preparing a composition for improving the expression of AQP3 in HaCaT cells, belonging to the technical field of HMO(s) application and the technical field of cosmetics. Background Art

[0002] When the stratum corneum of the skin lacks water, the normal metabolism of keratinocytes will be abnormal, resulting in a series of problems such as dryness, roughness, reduced elasticity, dullness, tightness and discomfort. The purpose of moisturizing cosmetics is to lock in moisture in the skin, keep the skin smooth and hydrated, and provide the skin with necessary nutrition and moisture.

[0003] Chinese patent 202110378551.1 discloses a moisturizing composition containing hydroxyethyl urea, which includes 0.2-12 parts of hydroxyethyl urea, 0.2-8 parts of betaine, 2-15 parts of butanediol, 0.6-5 parts of trehalose, 0.2-2 parts of 1,2-hexanediol, 36.5-94.78 parts of water, 0.02-0.5 parts of sodium hyaluronate, 2-8 parts of glycerol glucoside, and 2-5 parts of glycerol. The moisturizing compositions provided by patents 202211341383.X, 202210135924.7 and 201610312886.2 all contain the moisturizing ingredient hydroxyethyl urea. However, some groups may have allergic reactions to hydroxyethyl urea, with symptoms such as skin redness, swelling, and itching.

[0004] In summary, studying how to maintain the moisture content of the stratum corneum, improve the skin's ability to retain water, and maintain the skin's health is of great significance to the skin's ability to resist external stimuli. In the demand for product iteration, there is a need to provide the market with safe and excellent moisturizing products. Summary of the invention

[0005] Purpose of the invention: To provide a composition containing HMO(s) ingredients to meet people's demand for moisturizing products.

[0006] Technical solution: Human milk oligosaccharides (HMOs) are a unique mixture of oligosaccharides found in human milk. Among them, fucosyllactose, such as 2'-fucosyllactose (2'-FL), 3-fucosyllactose (3-FL), and lactose-N-tetraose (LNT), have been found to exhibit excellent biological functions in gastrointestinal barrier, immune enhancement, anti-inflammation, and promoting brain neural development.

[0007] The applicant surprisingly found that fucosyllactose has particular advantages in preparing a product for increasing the expression of AQP3 in HaCaT cells.

[0008] AQP3 (Aquaporin 3) is a substance on the cell membrane. It is a family of membrane channel proteins that are highly selective in transporting water. Their presence allows cells to quickly adjust their own volume and internal osmotic pressure, and plays an important role in maintaining the body's water balance. AQP3 is mainly expressed in keratinocytes and skin fibroblasts. It is not only involved in skin hydration and barrier function, but also plays an important role in skin damage, repair, and healing. It is an important guarantee for maintaining normal skin morphology and function.

[0009] In a first aspect, the present application provides the use of fucosyllactose in preparing a composition for increasing the expression of AQP3 in HaCaT cells.

[0010] Preferably, the fucosyllactose includes but is not limited to 2'-fucosyllactose (2'-fucosyllactose, 2'-FL), 3-fucosyllactose (3-FL), difucosyllactose (Difucosyllactose, DFL), lactose-N-fucopentaose I (Lacto-N-fucopentaose I, LNFP I), lactose-N-difucosyllactose I

[0011] (Lacto-N-difucohexaose I, LNDFH I).

[0012] In a second aspect, the present application provides a composition containing HMO, which contains algae syllactose.

[0013] Preferably, the fucosyllactose is selected from 2'-fucosyllactose (2'-fucosyllactose, 2'-FL), 3-fucosyllactose (3-FL), difucosyllactose (DFL), lactose-N-fucopentaose I (Lacto-N-fucopentaose I, LNFP I), lactose-N-difucosyllactose I

[0014] (Lacto-N-difucohexaose I, LNDFH I) or more thereof.

[0015] Preferably, the composition includes but is not limited to skin care lotion, skin care cream, hand cream, eye cream, lipstick and other cosmetics. Other raw materials and auxiliary materials of these cosmetics can be formulated according to the existing technology without special restrictions. There is no special restriction on the production method, and it can be produced according to conventional methods in the industry.

[0016] Preferably, the amount of fucosyllactose in the composition ranges from 0.03 to 2%.

[0017] In a third aspect, the present application provides a composition containing multiple HMOs, which contains fucosyllactose and lacto-N-tetraose (lacto-N-tetraose, LNT) and / or lacto-N-neotetraose (lacto-N-neotetraose, LNnT).

[0018] Preferably, the fucosyllactose is selected from 2'-fucosyllactose (2'-fucosyllactose, 2'-FL), 3-fucosyllactose (3-FL), difucosyllactose (DFL), lactose-N-fucopentaose I (Lacto-N-fucopentaose I, LNFP I), lactose-N-difucosyllactose I

[0019] (Lacto-N-difucohexaose I, LNDFH I) or more thereof.

[0020] The composition has excellent performance in moisturizing.

[0021] The applicant surprisingly found that in the composition, when the mass ratio of fucosyllactose to lactose-N-tetraose and / or lactose-N-neotetraose was 1:0.6-2, a synergistic moisturizing effect was produced.

[0022] Preferably, in the composition containing HMOs, the mass ratio of fucosyllactose to lactose-N-tetraose and / or lactose-N-neotetraose is 1:0.9-1.7.

[0023] Preferably, in the HMOs composition, the mass ratio of fucosyllactose to lactose-N-tetraose and / or lactose-N-neotetraose is 1:1.2-1.6.

[0024] Preferably, the composition includes but is not limited to skin care lotion, skin care cream, hand cream, eye cream, lipstick and the like.

[0025] Preferably, the composition may be a skin care lotion.

[0026] The composition of the present invention can be formulated into any suitable product form. Such product forms include, but are not limited to, aerosol sprays, creams, emulsions, solids, liquids, dispersions, foams, gels, lotions, mousses, ointments, powders, patches, pomades, solutions, hand-pump sprays, sticks, facial masks, etc. The composition of the present invention can be conveniently used to prepare or as a cosmetic, dermatological or pharmaceutical topical application product by various methods well known in the art.

[0027] The composition described in the present application may also be added with one or more of the following ingredients as needed: antimicrobial agents, antioxidants, chelating agents, colorants, depigmenting agents, emollients, emulsifiers, exfoliating agents, film formers, fragrances, moisturizers, insect repellents, lubricants, pharmaceutically active agents, moisturizers, photostabilizers, preservatives, skin care agents, skin penetration enhancers, sunscreens, stabilizers, surfactants, thickeners, viscosity regulators, vitamins or any combination thereof.

[0028] Preferably, the present invention provides a skin care lotion, which contains 1-5% glycerin, 3-10% 1,3-propylene glycol, 0.1-0.5% sodium polyacrylate, 2-5% methyl glucose sesquistearate, 2-5% PEG-20 methyl glucose sesquistearate, 2-5% cetearyl alcohol, 2-5% isohexadecane, 0.7-2.2% jojoba oil, 2-8% cetearyl ethylhexanoate, and 0.03-2% fucosyllactose.

[0029] Preferably, the skin care lotion contains 2-4% glycerin, 4-8% 1,3-propylene glycol, 0.2-0.4% sodium polyacrylate, 3-4% methyl glucose sesquistearate, 3-4% PEG-20 methyl glucose sesquistearate, 3-4% cetearyl alcohol, 3-4% isohexadecane, 0.9-1,8% jojoba oil, 3-6% cetearyl ethylhexanoate, and 0.05-2% fucosyllactose.

[0030] The skin care lotion contains 1-5% glycerol, 3-10% 1,3-propylene glycol, 0.1-0.5% sodium polyacrylate, 2-5% methyl glucose sesquistearate, 2-5% PEG-20 methyl glucose sesquistearate, 2-5% cetearyl alcohol, 2-5% isohexadecane, 0.7-2.2% jojoba oil, 2-8% cetearyl ethylhexanoate, 0.05-2% fucosyllactose, and further contains lactose-N-tetraose and / or lactose-N-neotetraose, wherein the amount of lactose-N-tetraose and / or lactose-N-neotetraose is 0.6-2 times that of fucosyllactose.

[0031] Preferably, the skin care lotion contains 2-4% glycerol, 4-8% 1,3-propylene glycol, 0.2-0.4% sodium polyacrylate, 3-4% methyl glucose sesquistearate, 3-4% PEG-20 methyl glucose sesquistearate, 3-4% cetearyl alcohol, 3-4% isohexadecane, 0.9-1,8% jojoba oil, 3-6% cetearyl ethylhexanoate, 0.03-2% fucosyllactose, and also contains lactose-N-tetraose and / or lactose-N-neotetraose, and the amount of lactose-N-tetraose and / or lactose-N-neotetraose is 0.9-1.7 times that of fucosyllactose.

[0032] Preferably, the skin care lotion contains 2-4% glycerol, 4-8% 1,3-propylene glycol, 0.2-0.4% sodium polyacrylate, 3-4% methyl glucose sesquistearate, 3-4% PEG-20 methyl glucose sesquistearate, 3-4% cetearyl alcohol, 3-4% isohexadecane, 0.9-1,8% jojoba oil, 3-6% cetearyl ethylhexanoate, 0.03-2% fucosyllactose, and also contains lactose-N-tetraose and / or lactose-N-neotetraose, and the amount of lactose-N-tetraose and / or lactose-N-neotetraose is 1.0-1.5 times that of fucosyllactose.

[0033] As mentioned above the percentages are by mass.

[0034] The moisturizing effect described in the present application can be tested by establishing a cell model using human keratinocytes or fibroblasts cultured in vitro. The effect of the sample on skin cells can be studied by observing the state of the cultured cells before and after the cells are exposed to the test substance and detecting the expression of proteins related to moisturizing.

[0035] The moisturizing effect of the skin care lotion described in this application is generally tested by a method that includes testing the moisture content of the stratum corneum of volunteers within a certain period of time before and after using the skin care lotion to determine the moisturizing effect of the skin care lotion. During the testing process, professional skin efficacy testing instruments are used to test the volunteers.

[0036] In a specific test of the long-lasting moisturizing effect, after using the skin care lotion described in the present application, the moisture content of the stratum corneum still increases 8 hours later, showing good durability.

[0037] In a fourth aspect, the present application provides a method for preparing a skin care lotion containing HMOs.

[0038] The preparation method of the skin care lotion comprises the following steps:

[0039] Step 1. Add the formulated amount of propylene glycol, sodium polyacrylate, methyl glucose sesquistearate, fucosyllactose, and / or lactose-N-tetraose and / or lactose-N-neotetraose to one-half the formulated amount of water, heat to 50-60° C., and stir until uniform.

[0040] Step 2. Evenly mix the formulated amounts of glycerin, PEG-20 methyl glucose sesquistearate, cetearyl alcohol, isohexadecane, jojoba oil, and cetearyl ethylhexanoate, heat to 60-70°C, and stir until uniform.

[0041] Step 3. Under stirring, mix the mixture obtained in step 1 and step 2 evenly to obtain the skin care lotion of the present invention.

[0042] Beneficial effects:

[0043] The present invention provides a composition with a moisturizing function. The composition has an excellent moisturizing effect, is safe and reliable, is non-irritating to the skin, and does not produce an allergic reaction. Using the composition of the present application can keep the water content of the stratum corneum of the skin sufficient, and the skin will be soft, smooth, delicate and elastic, thereby improving the water retention capacity of the skin and maintaining skin health. DETAILED DESCRIPTION

[0044] The present invention will be further described below in conjunction with specific examples. However, it should be understood that these examples are only used to illustrate the present invention and do not constitute a limitation of the scope of the present invention. The test methods in the following examples that do not specify specific conditions are usually based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, all percentages are weight percentages.

[0045] In the examples, fucosyllactose and lactose-N-tetraose and / or lactose-N-neotetraose were prepared by Shandong Henglu Biotechnology Co., Ltd., and other raw materials were purchased through commercial channels.

[0046] Example 1-37. Preparation of HMO aqueous solution composition

[0047] According to the concentration indicators in Table 1, the compositions of Examples 1 to 37 were prepared respectively, and the solvent was purified water.

[0048] Table 1.

[0049]

[0050]

[0051]

[0052] Example 38. Experimental study on the soothing efficacy of the solution described in Table 1.

[0053] 1 Test Principle

[0054] Aquaporins (AQPs) are integral membrane proteins that form water channels throughout biological membranes. The main function of AQPs is to transport water, glycerol, and other small solute molecules through biological membranes. Among them, AQP3 is highly expressed in the plasma membrane of keratinocytes and acts as a water and glycerol transporter in the basal layer of the epidermis of the skin. The glycerol content and water content of the epidermis of the skin without AQP3 will decrease, so in theory, an effective way to replenish water is to activate or increase the number of aquaporins to transport water from the dermis to the epidermis.

[0055] 2. Purpose of the experiment

[0056] The samples were added to human immortalized keratinocytes (HaCaT) for culture, and the effects of the samples on the expression of AQP3 in HaCaT cells were detected by immunofluorescence cytochemistry to indicate whether the samples had moisturizing effects.

[0057] 3. Test reagents

[0058] 3.1 Experimental cells

[0059] Human immortalized keratinocytes (HaCaT): Kunming Cell Bank, Chinese Academy of Sciences.

[0060] 3.2 Test reagents, see Table 2.

[0061] Table 2

[0062]

[0063]

[0064] 4. Immunofluorescence Detection

[0065] (1) Take out the sterile cell slide and place it in the culture plate.

[0066] (2) Collect the HaCaT cell suspension with a confluence of 80% to 90% in a T25 flask, count the cells, adjust the cell density, inoculate them on a culture plate, and culture them in a carbon dioxide incubator.

[0067] (3) Remove the culture plate, discard the culture medium in the wells, add working solutions of different concentrations according to the experimental group list, and continue culturing in a carbon dioxide incubator.

[0068] (4) Remove the culture plate, discard the culture medium in the wells, and add PBS for washing.

[0069] (5) Add 4% paraformaldehyde to fix the cells and then add PBS to wash.

[0070] (6) Add 0.2% Triton X-100 solution for permeabilization and add PBS for washing.

[0071] (7) Add bovine serum albumin solution for sealing.

[0072] (8) Anti-AQP3 Antibody and Goat Anti-Rabbit IgG H&L (Alexa Fluor 488) for staining and incubation, and then washed with PBS.

[0073] (9) Re-stain with DAPI staining solution and wash with PBS.

[0074] (10) Add a drop of anti-fluorescence quencher to the slide and seal the slide. Observe and take pictures under a fluorescence microscope.

[0075] 5. Calculation and determination of immunofluorescence test results

[0076] ImageJ 1.53t software was used to measure the fluorescence intensity, calculate the corrected total cell fluorescence, and calculate the increase rate of AQP3 expression in HaCaT cells compared with the blank control group. The results are recorded in Table 3.

[0077] Table 3. Analysis data of immunofluorescence cytochemistry staining results

[0078]

[0079]

[0080] Description: Positive control group (PC), EGF.

[0081] As the above data shows:

[0082] (1) Fucosyllactose can increase the expression of AQP3 in HaCaT cells and has a moisturizing effect, which can play a role in skin moisturizing.

[0083] (2) The combination of fucosyllactose with lacto-N-tetraose and / or lacto-N-neotetraose produced a synergistic effect in increasing the expression of AQP3 in HaCaT cells.

[0084] Examples 39-45. According to the formula described in Table 4 and the preparation method described in the technical solution, 100g of skin care lotion was prepared respectively.

[0085] Table 4

[0086]

[0087]

[0088] Control Examples 1-5. According to the formula described in Table 5 and the preparation method described in the technical solution, 100 g of skin care lotion was prepared respectively.

[0089] Table 5

[0090]

[0091]

[0092] Example 46. Test on the effects of the skin care lotions of Examples 39-45 and the skin care products of Control Examples 1-5 after application.

[0093] Healthy volunteers (female) aged between 25 and 35 were selected and randomly assigned to a group of 5. They were asked to try the skin care lotions of Examples 39 to 45 and the skin care products of Control Examples 1 to 5, and were respectively counted as Groups 39 to 45 of Examples and Groups 1 to 5 of Control Examples.

[0094] The facial skin of volunteers in each group was cleaned in the same way, and only 5% glycerol aqueous solution was sprayed on the day before the experiment, without any other skin care products. After the experiment began, each volunteer applied the skin care lotion provided by the group twice a day (once in the morning and once in the evening after washing), evenly applied to the facial skin, and no other skin care products were used during the period. The application was continued for 5 days, and the water content of the cheek skin (outer layer of the corneum) was measured using a skin moisture tester at 0h, 8h, 24h, 48h, and 120h, and the average value was calculated. The results are summarized in Tables 6 and 7.

[0095] Table 6. Test results of skin care lotion application effect prepared in Examples 39-45 and Comparative Examples 1-5

[0096]

[0097] Note: During the experiment, the volunteers in the embodiment group and the control group did not experience skin redness or swelling.

[0098] The above description is only the preferred embodiment of the present application and is not intended to limit the present application. Although the present application is described in detail with reference to the aforementioned embodiments, those skilled in the art can still modify the technical solutions described in the aforementioned embodiments or replace some of the technical features therein by equivalents. Any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the present application shall be included in the protection scope of the present application.

Claims

1. Application of fucosyllactose in the preparation of a composition for increasing the expression of AQP3 in HaCaT cells.

2. A composition containing HMO, characterized in that: Contains algae-syl lactose, Preferably, the fucosyllactose is selected from one or more of 2'-fucosyllactose (2'-fucosyllactose, 2'-FL), 3-fucosyllactose (3-FL), difucosyllactose (DFL), lactose-N-fucopentaose I (Lacto-N-fucopentaose I, LNFP I), and lactose-N-difucohexaose I (Lacto-N-difucohexaose I, LNDFH I), Preferably, the amount of fucosyllactose in the composition ranges from 0.05 to 2%.

3. The composition according to claim 2, characterized in that It also contains lacto-N-tetraose (LNT) and / or lacto-N-neotetraose (LNnT). Preferably, the mass ratio of fucosyllactose to lactose-N-tetraose and / or lactose-N-neotetraose in the composition is 1:0.6-2.

4. The composition according to claim 3, characterized in that The mass ratio of fucosyllactose to lactose-N-tetraose and / or lactose-N-neotetraose is 1:0.9-1.

7.

5. The composition according to claim 3, characterized in that The mass ratio of fucosyllactose to lactose-N-tetraose and / or lactose-N-neotetraose is 1:1.2-1.

6.

6. A skin care lotion, characterized in that: Contains 1-5% glycerol, 3-10% 1,3-propylene glycol, 0.1-0.5% sodium polyacrylate, 2-5% methyl glucose sesquistearate, 2-5% PEG-20 methyl glucose sesquistearate, 2-5% cetearyl alcohol, 2-5% isohexadecane, 0.7-2.2% jojoba oil, 2-8% cetearyl ethylhexanoate, 0.05-2% fucosyllactose, and the balance is water. The percentages are by mass.

7. The skin care lotion according to claim 6, characterized in that: Contains glycerin 2-4%, 1,3-propylene glycol 4-8%, sodium polyacrylate 0.2-0.4%, methyl glucose sesquistearate 3-4%, PEG-20 methyl glucose sesquistearate 3-4%, cetearyl alcohol 3-4%, isohexadecane 3-4%, jojoba oil 0.9-1,8%, cetearyl ethylhexanoate 3-6%, fucosyllactose 0.08-2%.

8. The skin care lotion according to claim 6, characterized in that: It contains 1-5% glycerin, 3-10% 1,3-propylene glycol, 0.1-0.5% sodium polyacrylate, 2-5% methyl glucose sesquistearate, 2-5% PEG-20 methyl glucose sesquistearate, 2-5% cetearyl alcohol, 2-5% isohexadecane, 0.7-2.2% jojoba oil, 2-8% cetearyl ethylhexanoate, 0.05-2% fucosyllactose, and also contains lactose-N-tetraose and / or lactose-N-neotetraose, the amount of lactose-N-tetraose and / or lactose-N-neotetraose is 0.6-2 times that of fucosyllactose.

9. The skin care lotion according to claim 6, characterized in that: It contains 2-4% glycerol, 4-8% 1,3-propylene glycol, 0.2-0.4% sodium polyacrylate, 3-4% methyl glucose sesquistearate, 3-4% PEG-20 methyl glucose sesquistearate, 3-4% cetearyl alcohol, 3-4% isohexadecane, 0.9-1,8% jojoba oil, 3-6% cetearyl ethylhexanoate, 0.08-2% fucosyllactose, and also contains lactose-N-tetraose and / or lactose-N-neotetraose, the amount of lactose-N-tetraose and / or lactose-N-neotetraose being 0.9-1.7 times that of fucosyllactose.

10. The skin care lotion according to claim 6, characterized in that: It contains 2-4% glycerol, 4-8% 1,3-propylene glycol, 0.2-0.4% sodium polyacrylate, 3-4% methyl glucose sesquistearate, 3-4% PEG-20 methyl glucose sesquistearate, 3-4% cetearyl alcohol, 3-4% isohexadecane, 0.9-1,8% jojoba oil, 3-6% cetearyl ethylhexanoate, 0.08-2% fucosyllactose, and also contains lactose-N-tetraose and / or lactose-N-neotetraose, the amount of lactose-N-tetraose and / or lactose-N-neotetraose being 1.0-1.5 times that of fucosyllactose.

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