Composition containing HMO

Abstract

The invention relates to a novel application of HMOs, in particular to an application of HMO in preparation of an antioxidant and whitening composition, further relates to an antioxidant and whitening skin care composition, and belongs to the technical field of application of HMOs. According to the technical scheme, the invention relates to the application of lacto-N-triose II (lacto-N-triose II, LNT II) in the aspect of preparing the antioxidant and / or whitening composition. The invention further provides a composition with the antioxidant and / or whitening functions, the composition contains the lactose-N-trisaccharide, and preferably, the composition contains 0.8%-6% of the lactose-N-trisaccharide (LNT II). The invention provides an application of LNTII in the aspects of oxidation resistance and whitening, and provides a stable body lotion composition. According to the composition disclosed by the invention, due to the use of the LNTII, the use of other whitening compounds is reduced or avoided, and adverse reactions such as sensitization and the like can be avoided.

Description

TECHNICAL FIELD

[0001] The present application relates to a new use of HMOs, in particular, to the application of HMOs in the preparation of a composition with antioxidant and / or whitening functions, and also to an antioxidant and whitening skin care composition, belonging to the technical field of HMOs application. BACKGROUND

[0002] Human milk oligosaccharides (HMOs) are a unique mixture of low molecular weight oligosaccharides present in human milk, and more than 200 have been identified. In recent years, with the in-depth study of the functions of HMOs, researchers have found that 2'-fucosyllactose (2'-FL), 3-fucosyllactose (3-FL), lacto-N-tetraose (LNT) and other HMOs have excellent biological functions in gastrointestinal barrier, improving digestive function, enhancing immune system, promoting brain neural development, etc., and have significant positive effects on the health of infants, adults and the elderly.

[0003] Recently, researchers have begun to explore the positive application of HMOs in the fields of beauty and cosmetics. CN202311644456.7 and KR101972925 both describe a composition containing 2'-FL, which has skin moisturizing and repairing effects. EP3067040 and KR102042967 both provide a composition containing 2'-FL, which can significantly increase the amount of collagen produced in cells and has skin anti-aging effects. CN202210733199.3 provides a composition containing multiple HMOs (such as 2'-fucosyllactose (2'-FL), 3-fucosyllactose (3-FL), lacto-N-tetraose (LNT), 3'-sialyllactose (3'-SL) or 6'-sialyllactose (6'-SL)), which has anti-wrinkle repair effects. In addition, WO2023025805 provides a composition containing at least one HMO selected from 2'-FL, difucosyllactose (DFL) and / or lacto-N-tetraose, for treating acne and combating skin inflammation.

[0004] In summary, it is known that 2'-fucosyllactose (2'-FL), 3-fucosyllactose (3-FL), lacto-N-tetraose (LNT), 3'-sialyllactose (3'-SL) or 6'-sialyllactose (6'-SL) and other components in HMOs have excellent biological functions in gastrointestinal barrier, improving digestive function, enhancing immune system, promoting brain neural development, etc. To expand the application of HMOs, it is necessary to further explore the new functions of the components of HMOs. SUMMARY

[0005] The present application provides an antioxidant or whitening composition containing HMOs.

[0006] Technical solution:

[0007] In a first aspect, the present application provides the use of lacto-N-triose II (LNT II) in the preparation of an antioxidant and / or whitening composition.

[0008] The antioxidant composition includes, but is not limited to, an antioxidant functional food or an antioxidant functional cosmetic. The antioxidant function includes, but is not limited to, a free radical scavenging function.

[0009] The whitening composition includes, but is not limited to, a whitening functional cosmetic. The whitening function includes, but is not limited to, a function of inhibiting tyrosinase activity.

[0010] The LNT II described in the present application can be prepared according to the prior art, such as the technology disclosed in the prior application of the applicant with application number 202211453172.5.

[0011] The antioxidant functional food can be prepared according to conventional functional food preparation technology.

[0012] The antioxidant whitening composition can be prepared according to conventional cosmetic preparation technology.

[0013] Free radicals are the main factors that cause the oxidation and deterioration of oil-based products and cause oxidative stress damage to the body, so antioxidants that prevent and scavenge free radicals are one of the most important means to prevent food deterioration and prevent and treat oxidative damage-related diseases.

[0014] Skin is a barrier protective organ that is directly exposed to the external environment, and skin exposed to the external environment for a long time is easily affected by natural environments such as ultraviolet light, leading to skin aging. The mechanism of skin aging is widely recognized as the free radical theory proposed by D. Harman in the 1950s. Free radicals are highly reactive molecules with unpaired electrons that can directly react with various cell membranes, lipids, proteins, DNA, RNA, etc., and cause damage to these substances, thereby causing problems such as skin wrinkles, skin cancer, atopic dermatitis, allergic dermatitis, etc.

[0015] Studies have shown that tea polyphenols, pomegranate polyphenols, and other substances can protect the skin from damage caused by oxidative stress by scavenging free radicals and inhibiting oxidation reactions, solving the problem of skin aging caused by free radicals. Tea polyphenols, pomegranate polyphenols, and other substances have a certain stimulating effect on the skin, and are not suitable for long-term use or for sensitive skin.

[0016] Applicants surprisingly found that lacto-N-triose, one of the components of HMOs, has free radical scavenging activity and can be prepared into an antioxidant composition, including but not limited to an antioxidant functional food or an antioxidant cosmetic.

[0017] Applicants also found that lacto-N-triose II (LNT II) has the ability to inhibit tyrosinase activity.

[0018] Traditional whitening ingredients include kojic acid, hydroquinone, fruit acid, tea polyphenols, pomegranate polyphenols, etc. Although these ingredients have certain whitening effects, kojic acid and hydroquinone are easily decomposed by light and heat, and kojic acid has been listed as a carcinogen of type 3 by the International Agency for Research on Cancer of the World Health Organization in 2017. Hydroquinone drugs have certain effects on the nervous system, and may cause dizziness, nausea, lethargy, etc. In addition, some people are allergic to hydroquinone substances, and red rashes may appear after application; fruit acid is an acidic substance extracted from fruits, which may cause irritation to the skin mucosa after being applied to the skin, and may also cause local redness or burning sensation, especially for sensitive skin. Long-term use of tea polyphenols and pomegranate polyphenols may cause irritation to the skin mucosa, leading to local redness or allergy. The discovery of lacto-N-triose II (LNT II) having the ability to inhibit tyrosinase activity provides a new safe and reliable material for the beauty industry and solves the shortcomings of traditional whitening ingredients.

[0019] In a second aspect, the present application provides a composition having antioxidant and / or whitening functions, which contains lacto-N-triose.

[0020] Preferably, the composition contains 0.8-6% of lacto-N-triose (LNT II).

[0021] When the amount of lacto-N-triose exceeds 6%, the antioxidant capacity is not positively correlated.

[0022] Stability experiments found that the antioxidant activity and the ability to inhibit tyrosinase activity of the skin cream containing lacto-N-triose (LNT II) showed a downward trend during storage. The researchers invented the technical solution of the third aspect of the present application.

[0023] In a third aspect, the present application provides a stable composition having antioxidant and / or whitening functions, which contains lacto-N-triose (LNT II) and lacto-N-tetraose (LNT).

[0024] Preferably, the mass ratio of lactose-N-triose (LNT II) and lactose-N-tetrose (LNT) in the composition is 1:1-2. When the amount of LNT exceeds 2 times the amount of LNT II, there is no difference in effect compared with 2 times.

[0025] Preferably, the mass ratio of lactose-N-triose (LNT II) and lactose-N-tetrose (LNT) in the composition is 1:1.2-1.8.

[0026] Further preferably, the mass ratio of lactose-N-triose (LNT II) and lactose-N-tetrose (LNT) in the composition is 1:1.5.

[0027] It has also been found that the reasonable combination of lactose-N-triose (LNT II) and lactose-N-tetrose (LNT) in the composition increases the moisturizing effect of the composition. The use of LNT can reduce the use of other moisturizing substances in the emollient cream.

[0028] The composition containing lactose-N-triose with antioxidant and / or whitening function according to the second or third aspect of the application has a dosage form of one of a cosmetic water, an emulsion, a paste, a cream, a mask, a capsule, a powder and a microneedle.

[0029] The composition with antioxidant and / or whitening function, optionally, further comprises cosmetically acceptable auxiliary agents. These auxiliary agents specifically include surfactants, emulsifiers, thickening agents and oils.

[0030] The suitable surfactants, emulsifiers, thickening agents and oils are well known to those skilled in the art.

[0031] The whitening composition according to the application, optionally, can further comprise other conventional cosmetic additives, such as fatty substances / oils, water, softening agents, antifoaming agents, etc.

[0032] In one embodiment, the composition containing lactose-N-triose with antioxidant and / or whitening function according to the application is an example of an emollient cream composition.

[0033] The emollient cream composition further contains oil, moisturizing agent and emulsifier.

[0034] The oil is not particularly limited and can be one or more of commonly used oils in the industry, such as caprylic / capric triglyceride, cetyl alcohol ethylhexanoate and hydrogenated polyisobutylene. The emulsion includes 5-10% of oil, preferably 6-8%, by mass percentage.

[0035] The emulsifier is not particularly limited and can be one or more of commonly used emulsifiers in the industry, such as glycerin stearate citrate, glycerin stearate SE (self-emulsifying), stearic acid, stearate, polyglyceryl-3-methylglucose distearate, etc. The emulsion includes 8-16% of the emulsifier, preferably 10-15%, in terms of mass percentage.

[0036] When the formulation does not contain LNT, the moisturizing agent includes, but is not limited to, one or more of sodium hyaluronate, butylene glycol, and trehalose. When the moisturizing agent includes two or more of the above specific substances, the present application does not have a special limitation on the ratio of the specific substances as long as the amount of the moisturizing agent can be met. The moisturizing emulsion provided by the present application includes 3-8% of the moisturizing agent, preferably 4-7%, in terms of mass percentage.

[0037] When the formulation contains LNT, the amount of the moisturizing agent is 2-5%, and the amount of the moisturizing agent is negatively correlated with the amount of LNT.

[0038] In one embodiment, the emulsion provided by the present application can further include a skin conditioning agent, which preferably includes allantoin and / or DL-panthenol. The amount of the skin conditioning agent is 0.6-1.3% of the skin conditioning agent, preferably 0.9-1.1%.

[0039] In one embodiment, optionally, the skin care emulsion provided by the present application further includes carbomer, the amount of which is 0.2-0.8%, preferably 0.3-0.5%.

[0040] In one embodiment, optionally, the skin care emulsion provided by the present application further includes ethylenediaminetetraacetic acid disodium salt, the amount of which is 0.01-0.1%, preferably 0.06-0.08%.

[0041] The percentages mentioned above are mass percentages.

[0042] The present application has the following beneficial effects over the prior art:

[0043] The present application provides the application of LNTII in antioxidation and whitening. And a stable skin care emulsion composition is provided. The use of LNTII in the composition provided by the present application reduces or avoids the use of other whitening compounds, which can avoid the occurrence of adverse conditions such as sensitization. The cosmetic product containing LNTII provided by the present application is safe for the human body, non-irritating, and stable to light and heat, and can be used for non-diagnostic treatment purposes of skin whitening. DETAILED DESCRIPTION

[0044] In the following examples, the experimental methods used are conventional methods unless otherwise specified; all materials, reagents, etc. can be obtained from commercial channels unless otherwise specified.

[0045] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. Generally, the nomenclature used herein is that used in the art.

[0046] It is to be understood that the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. It is further noted that the singular forms "a," "an," and "the" are used in combination with the term "comprising" or "containing" to refer to the open-ended term "comprising." It is to be understood that the terms "comprising" and "containing" are used in their broadest sense to mean that the described features, steps, operations, devices, components, and / or combinations thereof are included, but not limited to, the features, steps, operations, devices, components, and / or combinations thereof. It is to be understood that the terms "comprising" and "containing" are used in their broadest sense to mean that the described features, steps, operations, devices, components, and / or combinations thereof are included, but not limited to, the features, steps, operations, devices, components, and / or combinations thereof.

[0047] In the examples, LNT II and LNT were prepared by Shandong Henglu Biotechnology Co., Ltd., and other raw materials were purchased through commercial channels.

[0048] Example 1. LNT II hydroxyl radical scavenging experiment

[0049] 1. Solution preparation:

[0050] (1) Prepare a lacto-N-triose (LNT II) aqueous solution with a concentration of 6 mg / mL according to the conventional method.

[0051] (2) Phosphate (PBS) buffer preparation: prepare a phosphate buffer with pH 7.4 according to the conventional method.

[0052] 2. Hydroxyl radical scavenging experiment scheme

[0053] (1) Blank test: in the reaction system, add distilled water 150 μL or 200 μL, 1.5 mM FeSO4 solution 200 μL, 150 mM phosphate (PBS) buffer (pH 7.4) 300 μL, 0.75 mM o-diazenophenol solution 200 μL, mix well, add 0.1% (v / v) H2O2 solution 200 μL, mix well, react at 37°C for 30 min in the dark, and measure the absorbance at 536 nm wavelength, recorded as A0;

[0054] (2) LNT II radical scavenging test: in the reaction system, 150 μL or 200 μL of lacto-N-triose aqueous solution (LNT II, 6 mg / mL), 200 μL of 1.5 mM FeSO4 solution, 300 μL of 150 mM PBS buffer (pH 7.4), 200 μL of 0.75 mM 1, 10-phenanthroline solution were added in turn, mixed well, 200 μL of 0.1% (v / v) H2O2 solution was added, mixed well, reacted at 37°C for 30 min in the dark, the absorbance was measured at 536 nm wavelength, and recorded as A i ;

[0055] (3) Control test: in the reaction system, 350 μL or 400 μL of distilled water, 200 μL of 1.5 mM FeSO4 solution, 300 μL of 150 mM PBS buffer (pH 7.4), 200 μL of 0.75 mM 1, 10-phenanthroline solution were added in turn, mixed well, mixed well, reacted at 37°C for 30 min in the dark, the absorbance was measured at 536 nm wavelength, and the average value of three parallel samples was calculated, recorded as A j ;

[0056] According to the following formula, the hydroxyl radical scavenging rate was calculated, and the results are recorded in Table 1.

[0057]

[0058] Table 1.

[0059] Volume of lacto-N-triose aqueous solution (LNT II) Hydroxyl radical clearance, % 150 μL 29.86 200 μL 34.15

[0060] Example 2. Tyrosinase activity inhibition test (whitening performance study)

[0061] 1. Solution preparation

[0062] (1) Preparation of sodium hydrogen phosphate-citric acid buffer (pH 6.8):

[0063] a) Prepare 0.2 mol / L sodium hydrogen phosphate and 0.1 mol / L citric acid monohydrate aqueous solution;

[0064] b) Take 154.5 mL of sodium hydrogen phosphate aqueous solution and 45.5 mL of citric acid aqueous solution, mix well, adjust pH, and adjust pH to 6.8 with 0.2 mol / L sodium hydrogen phosphate or 0.1 mol / L citric acid.

[0065] (2) Preparation of tyrosinase solution: prepare 100 u / mL tyrosinase solution with the above sodium hydrogen phosphate-citric acid buffer (pH 6.8).

[0066] (3) L-dopa solution preparation: 1 mg / mL L-dopa solution was prepared using the above sodium phosphate dibasic-citric acid buffer (pH 6.8).

[0067] (4) Lactose-N-triose (LNT II) solution preparation: 2 mg / mL lactose-N-triose (LNT II) solution was prepared using the above sodium phosphate dibasic-citric acid buffer (pH 6.8).

[0068] 2. Tyrosinase activity inhibition experiment (Reference Example 1) TSHRH 015-2018 Cosmetics - Inhibition of tyrosinase activity test Method )

[0069] (1) 1 mL or 1.5 mL of lactose-N-triose solution (2 mg / mL) was taken, 0.5 mL of tyrosinase was added, mixed, and 10 min of 37°C water bath was performed, 2 mL of L-dopa was then added, and the reaction time was controlled for 5 min, and the absorbance T was measured at 475 nm.

[0070] (2) Sample background: 1 mL or 1.5 mL of lactose-N-triose solution was taken, 0.5 mL of sodium phosphate dibasic-citric acid buffer (pH 6.8) was added, mixed, and 10 min of 37°C water bath was performed, 2 mL of L-dopa was then added, and the reaction time was controlled for 5 min, and the absorbance T0 was measured at 475 nm.

[0071] (3) Enzyme reaction tube: 1 mL or 1.5 mL of sodium phosphate dibasic-citric acid buffer (pH 6.8) was taken, 0.5 mL of tyrosinase solution was added, mixed, and 10 min of 37°C water bath was performed, 2 mL of L-dopa was then added, and the reaction time was controlled for 5 min, and the absorbance C was measured at 475 nm.

[0072] (4) Solvent background: 1.5 mL or 2 mL of sodium phosphate dibasic-citric acid buffer (pH 6.8) was taken, mixed, and 10 min of 37°C water bath was performed, 2 mL of L-dopa was then added, and the reaction time was controlled for 5 min, and the absorbance C0 was immediately measured at 475 nm.

[0073] The tyrosinase activity inhibition rate was calculated according to the following formula, and the results are recorded in Table 2.

[0074]

[0075] Table 2

[0076] Volume of lacto-N-triose aqueous solution (LNT II) Inhibition of tyrosinase activity, % 1 mL 20.88 1.5 mL 26.77

[0077] Example 3-8, Comparative Example 1. Preparation of emollient cream composition: The emollient cream compositions of Examples 3-8 and Comparative Example 1 were prepared according to the formulations described in Table 3, respectively.

[0078] Table 3

[0079]

[0080] The preparation method is as follows:

[0081] (1) Dissolve sodium hyaluronate, DL-pantothenyl alcohol, disodium ethylenediaminetetraacetate, lactose-N-triose (LNT II) in 70% water in the formula to obtain an aqueous phase;

[0082] (2) Under rapid stirring, gradually add carbomer, caprylic triglyceride, glyceryl stearate SE in the formula to the aqueous phase obtained in step 1, and supplement the amount of water in the formula during stirring to obtain a skin lotion.

[0083] Example 9-14, Comparative Example 2. Preparation of stable skin lotion compositions: According to the formula described in Table 4, respectively, the skin lotion compositions of Examples 9-14 and Comparative Example 2 were prepared.

[0084] Table 4

[0085]

[0086] (1) Dissolve sodium hyaluronate, DL-pantothenyl alcohol, disodium ethylenediaminetetraacetate, lactose-N-triose, lactose-N-tetrose in 70% water in the formula to obtain an aqueous phase;

[0087] (2) Under rapid stirring, gradually add carbomer, caprylic triglyceride, glyceryl stearate SE in the formula to the aqueous phase obtained in step 1, and supplement the amount of water in the formula during stirring to obtain a skin lotion.

[0088] Example 11. Stability experiment of the skin lotion compositions obtained in Examples 3-14 and Comparative Examples 1-2.

[0089] Take 20 mL of the skin lotion composition obtained in Examples 3-14 or Comparative Examples 1-2, respectively, and place it in a ground-mouth reagent bottle, and place it in a constant-temperature and constant-humidity box at 50°C and a relative humidity of 60%, and respectively determine the hydroxyl radical scavenging activity and tyrosinase enzyme activity inhibition at 0 days, 30 days, and 60 days.

[0090] (I) Hydroxyl radical scavenging experiment of the skin lotion compositions obtained in Examples 3-14 and Comparative Examples 1-2.

[0091] According to the method of the hydroxyl radical scavenging experiment described in Example 1, respectively, use 150 μL of the skin lotion composition obtained in Examples 3-14 and Comparative Examples 1-2 instead of the 6 mg / mL aqueous solution of lactose-N-triose (LNT II) in Example 1 to perform the hydroxyl radical scavenging experiment. The results are recorded in Table 5.

[0092] Table 5

[0093]

[0094] The data in Table 5 show that: (1) the skin lotion compositions obtained in Examples 3-14 and Control Examples 1-2 all have good hydroxyl radical scavenging ability.

[0095] (2) the skin lotion composition of Example 3-8 has a small decrease in hydroxyl radical scavenging rate during the stability experiment.

[0096] (3) the skin lotion compositions of Examples 9-14, which are based on the formulation of the skin lotion composition of Example 3-8 and have added lacto-N-tetraose, have no change in hydroxyl radical scavenging rate with storage time.

[0097] (4) in Control Example 1, the amount of LNT II is 8%, which has a similar hydroxyl radical scavenging effect as the amount of LNT II of 6% (Example 8).

[0098] (5) in Control Example 2, the amount of LNT is 3 times that of LNT II, which has a similar stability effect as the amount of LNT II of 2 times (Example 9).

[0099] (B) Inhibition of tyrosinase activity experiment of the skin lotion compositions obtained in Examples 3-14 and Control Examples 1-2.

[0100] (1) 10 mL of one of the skin lotion compositions obtained in Examples 3-14 or Control Examples 1-2 was diluted to 20 mL with purified water.

[0101] (2) 10 mL of the solution obtained in (1) was diluted to 20 mL with purified water to obtain a sample solution for the inhibition of tyrosinase activity experiment.

[0102] According to the method for the inhibition of tyrosinase activity experiment described in Example 2, 1 mL of the sample solution prepared in (2) above was used instead of the 2 mg / mL aqueous solution of lacto-N-triose (LNT II) in Example 2 to perform the tyrosinase activity experiment. The results are recorded in Table 6.

[0103] Table 6

[0104]

[0105] The data in Table 6 show that:

[0106] (1) the skin lotion compositions obtained in Examples 3-14 and Control Examples 1-2 all have good tyrosinase activity inhibition activity.

[0107] (2) The tyrosinase activity inhibition rate of the emollient cream composition of Examples 3-8 showed a slight downward trend during the stability experiment.

[0108] (4) In Control Example 1, the amount of LNT II was 8%, which had a similar tyrosinase activity inhibition effect as the amount of LNT II of 6% (Example 8).

[0109] (5) In Control Example 2, the amount of LNT was 3 times that of LNT II, and the stability effect was basically the same as that of 2 times (Example 9).

[0110] The above only describes the preferred embodiments of the present application and is not intended to limit the present application. Although the present application has been described in detail with reference to the foregoing embodiments, modifications or equivalent replacements of the technical solutions described in the foregoing embodiments or some technical features thereof can still be made by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. Application of lactose-N-triose II (LNT II) in the preparation of antioxidant and / or skin-whitening compositions; Preferably, the antioxidant composition includes, but is not limited to, food products or cosmetics with antioxidant functions; Preferably, the antioxidant function includes, but is not limited to, free radical scavenging function; Preferably, the whitening composition includes, but is not limited to, cosmetics with whitening functions; Preferably, the whitening function includes, but is not limited to, the function of inhibiting tyrosinase activity.

2. The composition with antioxidant and / or whitening functions as described in claim 1, characterized in that, It contains lactose-N-trisaccharide.

3. The composition according to claim 2, characterized in that, It contains 0.8-6% lactose-N-trisaccharide by mass.

4. A stable composition having antioxidant and / or whitening functions, characterized in that, It contains lactose-N-trisaccharide (LNT II) and lactose-N-tetrasaccharide (LNT).

5. The composition according to claim 4, characterized in that, The mass ratio of lactose-N-trisaccharide (LNT II) to lactose-N-tetrasaccharide (LNT) in the composition is 1:1-2.

6. The composition according to claim 4, characterized in that, The mass ratio of lactose-N-trisaccharide (LNT II) to lactose-N-tetrasaccharide (LNT) in the composition is 1:1.2-1.

8.

7. The composition according to claim 4, characterized in that, The composition contains lactose-N-trisaccharide (LNT II) and lactose-N-tetrasaccharide (LNT) in a mass ratio of 1:1.

5.

8. The composition according to claims 1-7, characterized in that, The composition also contains one or more of caprylic / capric triglycerides, cetyl ethylhexanoate, and hydrogenated polyisobutylene. Preferably, the amount of caprylic / capric triglyceride, cetyl ethylhexanoate and hydrogenated polyisobutylene is in the range of 5-10% by mass percentage, preferably 6-8%.

9. The composition according to claims 1-7, characterized in that, The composition also contains one or more of the following: glyceryl stearate citrate, glyceryl stearate SE (self-emulsifying), stearic acid, stearate, polyglycerol-3-methylglucose distearate.

10. The composition according to claim 9, characterized in that, The composition contains glyceryl stearate citrate, glyceryl stearate SE (self-emulsifying), stearic acid, stearate, and polyglycerol-3-methylglucose distearate in an amount ranging from 8% to 16%, preferably 10% to 15%.

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