Penetration enhancing compositions based on polyglyceryl-10 laurate

Abstract

The application provides a penetration promoting composition based on polyglyceryl-10 laurate, which comprises ergothioneine and polyglyceryl-10 laurate, and the weight ratio of the ergothioneine and the polyglyceryl-10 laurate is 10:1 to 1:30, wherein the content of the polyglyceryl-10 laurate is 0.01wt%-3wt%. The penetration rate of the composition after 24h vertical transdermal diffusion is greater than or equal to 50%. The application further discloses application of the aqueous dispersion system in a skin external preparation.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of skin external agents, in particular to the directional penetration of ergothioneine in specific content by polyglyceryl-10 laurate, and more particularly to a penetration composition based on polyglyceryl-10 laurate, which can improve the penetration and absorption of ergothioneine in the skin, and has excellent stability. BACKGROUND

[0002] In 1909, Charles Tanret first isolated ergothioneine (EGT) from ergot, which has a structure of connecting three methyl groups to the amino group of histidine and a thiol group to the imidazole ring, and belongs to the derivative of histidine. Its structure is as follows: It is this unique molecular structure of ergothioneine that gives it super strong antioxidant properties. It can not only capture singlet oxygen, quench a variety of free radicals and chelate divalent metal ions to inhibit the pro-oxidation of these ions, but also activate antioxidant enzymes and inhibit superoxide dismutase to promote the clearance of free radicals, and additionally affect the oxidation of various heme proteins. Ergothioneine exists in almost every part of the human body, but humans cannot synthesize ergothioneine themselves and must obtain it from the outside world. There is a special protein in the human body: transport protein OCTN1 (organic cation transporter N1).

[0003] Ergothioneine is a water-soluble amino acid molecule with excellent water solubility. Due to the easy water solubility of ergothioneine, it can be transported through the biological membrane by transport protein OCTN1, thereby distributing throughout the body. In living organisms, the expression of transport protein OCTN1 is closely related to the level of free radicals.

[0004] Studies have shown that when cells are under oxidative stress or inflammation, the expression of SLC22A4 gene encoding OCTN1 will be up-regulated, so that more transport protein OCTN1 will be distributed on the cell membrane, prompting ergothioneine to enter the cell to clear excess free radicals. When the free radicals return to normal levels, the SLC22A4 gene will be silenced. Therefore, ergothioneine is considered to be the only natural antioxidant that can enter the cell and mitochondria and intelligently clear excess free radicals without affecting the normal function of living organisms.

[0005] In addition, due to the oxidation and reduction mechanism of glutathione in the body, ergothioneine has the ability to regenerate in the human body. For example, after quenching singlet oxygen, ergothioneine will become "hydroperoxy ergothioneine", and under the action of glutathione, it will return to the state of ergothioneine. This regenerative ability also gives ergothioneine the ability to persistently resist oxidation and clear free radicals.

[0006] With more and more researches on it at home and abroad, in addition to its functions of antioxidant and anti-free radical, it also has effects of anti-radiation, whitening, anti-aging, sun protection, reducing inflammation, reducing glycosylation, etc. Ergothioneine with so many functions and strong efficacy is not only popular in food and health products, but also more and more used in skin care products.

[0007] Ergothioneine in cosmetics is mainly absorbed by the skin, but the skin as the outermost protection of the human body has the functions of resistance and regulation. The special structure of the skin leads to its oil-loving nature. The hydrophilic nature of ergothioneine makes it difficult to be absorbed by the skin.

[0008] Chinese patent application CN116059153B discloses a milky white essence containing Schizochytrium limacinum fermentation product and a preparation method thereof, which contains Schizochytrium limacinum fermentation product filtrate, Schizochytrium limacinum fermentation product cytolysis product, ergothioneine, skin probiotic, acetylchitosamine, polyglycerol 10 laurate, citric acid fatty acid glyceride, emollient, sodium hyaluronate and polyhydric alcohol. The milky white essence containing oil phase and water phase has excellent stability, excellent absorption effect, good spreadability, low solubilizer content, and also has excellent antioxidant and anti-aging and moisturizing effects. Although the application simultaneously uses ergothioneine and polyglycerol 10 laurate, it does not suggest that polyglycerol-10 laurate has a penetration-promoting effect on ergothioneine, indicating that the two are simply added in a product.

[0009] Therefore, it is very meaningful to find a stable method that can increase the absorption of ergothioneine in the skin.

[0010] The present application unexpectedly finds that polyglycerol-10 laurate has a directional penetration-promoting effect on ergothioneine with a specific content, based on which the present application provides a penetration-promoting composition based on polyglycerol-10 laurate, which can improve the penetration and absorption of ergothioneine in the skin and has excellent stability. SUMMARY

[0011] Based on the directional penetration-promoting effect of polyglycerol-10 laurate on ergothioneine with a specific content, the present application provides a penetration-promoting composition based on polyglycerol-10 laurate, which can improve the penetration and absorption of ergothioneine in the skin and has excellent stability.

[0012] The present application provides a penetration-promoting composition based on polyglycerol-10 laurate, which contains ergothioneine and polyglycerol-10 laurate, and the weight ratio of the ergothioneine and polyglycerol-10 laurate is 10:1 to 1:30, wherein the content of the polyglycerol-10 laurate is 0.01wt%-3wt%.

[0013] In a preferred embodiment, the content of the polyglyceryl-10 laurate is 0.01wt%-2wt%.

[0014] In a preferred embodiment, the weight ratio of the ergothioneine and the polyglyceryl-10 laurate is 10:1 to 1:25.

[0015] In a preferred embodiment, the weight ratio of the ergothioneine and the polyglyceryl-10 laurate is 2:1 to 1:25.

[0016] In a preferred embodiment, the weight ratio of the ergothioneine and the polyglyceryl-10 laurate is 1:1 to 1:25.

[0017] In a preferred embodiment, the composition can further comprise ≥96wt% of a carrier suitable for use in the field of skin external agents.

[0018] In a preferred embodiment, the carrier is selected from the group consisting of water.

[0019] In a preferred embodiment, the composition has a permeation rate ≥50% after 24h vertical transdermal diffusion.

[0020] In a preferred embodiment, the composition has a permeation rate of 54.28-76.11% after 24h vertical transdermal diffusion.

[0021] The present application also provides the use of polyglyceryl-10 laurate for promoting the permeation of ergothioneine.

[0022] In a preferred embodiment, the weight ratio of the ergothioneine and the polyglyceryl-10 laurate is 10:1 to 1:30.

[0023] In a preferred embodiment, the weight ratio is 10:1 to 1:25.

[0024] In a further preferred embodiment, the weight ratio is 2:1 to 1:25.

[0025] In a most preferred embodiment, the weight ratio is 1:1 to 1:25.

[0026] The present application also provides the use of the penetration-promoting composition based on polyglyceryl-10 laurate in skin external agents.

[0027] In a preferred embodiment, the composition is used in skin external agents in an amount of 0.01wt%-20wt%.

[0028] In a preferred embodiment, the skin external agent is selected from the group consisting of creams, lotions, serums, gels, toners, masks, cleansers, bath agents, make-up removers, hair-washing and hair-care agents, oils, toilet waters, and the like.

[0029] The present application has the following advantages:

[0030] The ergothioneine combination obtained according to the above scheme can ensure very stability and increase the transdermal absorption of ergothioneine in the skin. BRIEF DESCRIPTION OF DRAWINGS

[0031] Figure 1 A working schematic diagram of a vertical diffusion cell used in the permeability test example;

[0032] Figure 2 A high performance liquid chromatogram of ergothioneine in Example 3 (3h);

[0033] Figure 3 A high performance liquid chromatogram of ergothioneine in Example 3 (24h). DETAILED DESCRIPTION

[0034] 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. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present application, the preferred methods and materials are described. For the purposes of the present application, the following terms are defined below.

[0035] To provide a more concise description, some of the numerical descriptions given herein are not modified by the term "about". It will be understood that whether or not the term "about" is expressly used, every quantity given herein is intended to refer to the actual given value, and also to approximations within experimental and / or measurement error, including the precision of the measurement and / or the limitations of the experimental and / or measurement techniques used.

[0036] To provide a more concise description, some of the numerical descriptions given herein are recited as a range from about X amount to about Y amount. It will be understood that when a range is recited, the range is not limited to the recited upper and lower amount, but rather includes the entire range of amounts from about X amount to about Y amount or any amount therebetween.

[0037] Carriers suitable for use in the field of topical skin preparations

[0038] The carrier suitable for the skin external agent of the present application is water, and the skin external agent can be selected from the group consisting of cream, lotion, essence, jelly, toner, pack, cleanser, bath agent, make-up remover, hair washing and conditioning agent, oil agent, floral water, and the like.

[0039] Permeation rate

[0040] The permeability of the present application is the permeability calculated from the amount of ergothioneine permeated through pigskin after 24h of vertical transdermal diffusion. In the present application, it is considered that the permeability is ≥ 50% after 24h of vertical transdermal diffusion experiment, and it is considered that it has a penetration promoting effect.

[0041] Hereinafter, the technical solutions of the present application will be described in detail through preferred embodiments, but the scope of the present application is not limited to these embodiments, which are intended to illustrate the technical solutions of the present application rather than limit the scope of the present application. The test methods in the following examples, unless otherwise specified, are generally carried out under conventional conditions, or under the conditions recommended by the manufacturer. Unless otherwise specified, all percentages and parts are by weight.

[0042] The experimental materials used in the embodiments of the present application are as follows:

[0043] • Ergothioneine (purchased from Shanghai Ergothioneine Biotechnology Group Co., Ltd.)

[0044] • Polyglyceryl-10 laurate (trade name NIKKOL Decaglyn 1-L, purchased from NIPPON SURFACTANT INDUSTRIES CO., LTD.)

[0045] • Deionized water (self-made)

[0046] • Neosolue TM • Aqulio Bis-diethoxydiglycol cyclohexane 1,4-dicarboxylate (purchased from Nippon Fine Chemical Co., Ltd.)

[0047] • 1,2-pentanediol (trade name: effisin pg multifunctional, purchased from Ashland LLC) TM

[0048] • Laurureth (purchased from TGI)

[0049] • RH40 (trade name RH 40, purchased from BASF)

[0050] • Polyglyceryl-10 myristate (trade name NIKKOL Decaglyn 1-M, purchased from LABORATOIRES SETHIC INNOVATIONS)

[0051] • Inulin lauryl carbamate (trade name INUTEC SL1, purchased from NIKKOL)

[0052] • 0.9wt% physiological saline (self-made)

[0053] Experimental instruments

[0054] • Fixed stirrer IKA RW20

[0055] • Weighing balance METTLER TOLEDO PL602-S

[0056] ​• Transdermal diffusion tester TK-12D Shanghai Yuren Scientific Instrument Co., Ltd.

[0057] • Contact pig skin, 1-2 month old small pig back skin Chongqing Meisheng Trading Co., Ltd.

[0058] • High performance liquid chromatograph with diode array detector Waters ARC HPLC

[0059] • Analytical balance METTLER TOLEDO XS205

[0060] • Digital ultrasonic cleaner Kunshan Ultrasonic Instrument Co., Ltd. KQ-800DE

[0061] • Constant temperature oven MMM Friocell 707

[0062] Examples 1-15: Preparation of ergothioneine and polyglyceryl-10 laurate different proportion concentration aqueous solution

[0063] Sample preparation: Ergothioneine and polyglyceryl-10 laurate were weighed according to the mass parts shown in Table 1, and after adding deionized water to make up 100 mass parts, they were kept stirring at room temperature until transparent liquid, ready for use.

[0064] Table 1: Addition amount of each component in ergothioneine and polyglyceryl-10 laurate different proportion concentration aqueous solution

[0065]

[0066] Test Example 1: Permeability test

[0067] Test sample: Ergothioneine and polyglyceryl-10 laurate different proportion concentration aqueous solution prepared in Examples 1-15

[0068] Test steps:

[0069] 1) Take out the frozen pig skin, after thawing, remove the subcutaneous fat and connective tissue with scissors.

[0070] 2) Soak the pig skin in 0.9wt% physiological saline for 1 hour.

[0071] 3) Open the vertical transdermal diffusion instrument as shown in Figure 1 , and heat in water bath (constant temperature 32℃).

[0072] 4) Put a magnetic stirrer in the receiving pool, then fix the cut pig skin between the supply pool and the receiving pool, with the epidermis layer upwards and the dermis layer in contact with the receiving pool. Add physiological saline as receiving liquid from the sampling tube into the receiving pool, and drain the air bubbles. Cover the supply pool with a cover glass, and put it into the transdermal diffusion device, constant temperature (32℃) stirring (440r / min) for 1h.

[0073] 5) Add each test sample to the feed tank, where the raw material is located, contact the pigskin, constant temperature (32℃), stirring (440r / min), after 3h, 24h, respectively from the sampling tube to receive liquid, to be measured.

[0074] The content of ergothioneine in the receiving liquid is determined by high performance liquid chromatography, and the cumulative penetration rate is calculated. The specific steps are as follows:

[0075] 6) The content of ergothioneine in the receiving liquid is determined by high performance liquid chromatography

[0076] a. Chromatographic conditions: the chromatographic column is Agilent HILIC Plus chromatographic column (4.6mm*100mm, 3.5μm), the mobile phase is 0.1% formic acid acetonitrile solution-0.1% formic acid water solution with a volume ratio of 80:20, the flow rate is 1mL / min, the column temperature is 30℃, the injection amount is 10μL, and the detection wavelength is 262nm.

[0077] b. Ergothioneine standard stock solution (0.5g / L) preparation: weigh 50mg of ergothioneine standard (accurate to 0.01mg), dissolve with water and dilute to 100mL. Store at 4℃.

[0078] c. Preparation of ergothioneine standard working solution: accurately pipette a certain amount of ergothioneine standard stock solution, and dilute with 0.1% formic acid methanol solution to obtain a series of standard working solutions (mass concentrations are 5μg / mL, 10μg / mL, 25μg / mL, 50μg / mL, 100μg / mL respectively).

[0079] d. Standard curve drawing: according to the above chromatographic conditions, the standard working solution is determined, and the mass concentration of the standard working solution is taken as the abscissa and the peak area is taken as the ordinate to draw the standard curve.

[0080] e. Sample treatment and determination: weigh 0.1-0.5g (accurate to 0.1mg) of sample in a 10mL colorimetric tube, add 8mL of 0.1% formic acid methanol solution, and ultrasonic extract for 20min. Dilute with 0.1% formic acid methanol solution, shake well, filter part of the liquid through a 0.22μm filter membrane, and determine according to the above chromatographic conditions. Quantify the sample on the standard curve. The response value of the analyte in the sample solution should be within the linear range of the standard curve.

[0081] 7) Calculate the cumulative penetration rate

[0082] EGT penetration rate = (m 取样量 ×C EGT ) / (m 上样量 ×C EGT总 )×100%

[0083] Table 2 Permeability of aqueous solutions of different proportions of ergothioneine and polyglyceryl-10 laurate

[0084]

[0085]

[0086] If the permeation rate value of the former is greater than that of the latter, it is considered to have a synergistic penetration effect; if the permeation rate value of the former is less than or equal to that of the latter, it is considered to have no synergistic penetration effect.

[0087] The composition sample prepared in Example 2 has a 3h permeation rate of 3.56%, which is less than the 4.12% permeation rate of the composition sample prepared in Example 1, and a 24h permeation rate of 24.56%, which is less than the 46.13% permeation rate of the composition sample prepared in Example 1, and it is considered that the composition sample prepared in Example 2 has no penetration effect;

[0088] The composition sample prepared in Example 3 has a 3h permeation rate of 16.93%, which is greater than the 4.12% permeation rate of the composition sample prepared in Example 1, and a 24h permeation rate of 65.42%, which is greater than the 46.13% permeation rate of the composition sample prepared in Example 1, and it is considered that the composition sample prepared in Example 3 has a synergistic penetration effect;

[0089] The composition sample prepared in Example 4 has a 3h permeation rate of 22.56%, which is greater than the 4.12% permeation rate of the composition sample prepared in Example 1, and a 24h permeation rate of 75.34%, which is greater than the 46.13% permeation rate of the composition sample prepared in Example 1, and it is considered that the composition sample prepared in Example 4 has a synergistic penetration effect;

[0090] The composition sample prepared in Example 5 has a 3h permeation rate of 26.64%, which is greater than the 4.12% permeation rate of the composition sample prepared in Example 1, and a 24h permeation rate of 76.11%, which is greater than the 46.13% permeation rate of the composition sample prepared in Example 1, and it is considered that the composition sample prepared in Example 5 has a synergistic penetration effect;

[0091] In summary, by comparing the 3h permeation rate with the 24h permeation rate, it is known that when the content of ergothioneine in the composition is 1wt%, the content of polyglyceryl-10 laurate is 0.5-2wt% (i.e. ergothioneine: polyglyceryl-10 laurate = 2:1, 1:1, 1:2), the composition exhibits synergistic penetration effect. When the content of polyglyceryl-10 laurate is 0.1wt% (i.e. ergothioneine: polyglyceryl-10 laurate = 10:1), the composition has no penetration effect.

[0092] The 3h permeation rate of the composition sample prepared in Example 7 is 10.05%, which is less than the 3h permeation rate of the composition sample prepared in Example 6, which is 10.46%, and the 24h permeation rate of the composition sample prepared in Example 7 is 39.24%, which is less than the 24h permeation rate of the composition sample prepared in Example 6, which is 47.01%, so it is considered that the composition sample prepared in Example 7 does not have synergistic penetration effect;

[0093] The 3h permeation rate of the composition sample prepared in Example 8 is 19.51%, which is greater than the 3h permeation rate of the composition sample prepared in Example 6, which is 10.46%, and the 24h permeation rate of the composition sample prepared in Example 8 is 54.28%, which is greater than the 24h permeation rate of the composition sample prepared in Example 6, which is 47.01%, so it is considered that the composition sample prepared in Example 8 has synergistic penetration effect;

[0094] The 3h permeation rate of the composition sample prepared in Example 9 is 26.31%, which is greater than the 3h permeation rate of the composition sample prepared in Example 6, which is 10.46%, and the 24h permeation rate of the composition sample prepared in Example 9 is 68.45%, which is greater than the 24h permeation rate of the composition sample prepared in Example 6, which is 47.01%, so it is considered that the composition sample prepared in Example 9 has synergistic penetration effect;

[0095] The 3h permeation rate of the composition sample prepared in Example 10 is 26.03%, which is greater than the 3h permeation rate of the composition sample prepared in Example 6, which is 10.46%, and the 24h permeation rate of the composition sample prepared in Example 10 is 67.49%, which is greater than the 24h permeation rate of the composition sample prepared in Example 6, which is 47.01%, so it is considered that the composition sample prepared in Example 10 has synergistic penetration effect;

[0096] The 3h permeation rate of the composition sample prepared in Example 11 is 9.84%, which is less than the 3h permeation rate of the composition sample prepared in Example 6, which is 10.46%, and the 24h permeation rate of the composition sample prepared in Example 11 is 36.32%, which is less than the 24h permeation rate of the composition sample prepared in Example 6, which is 47.01%, so it is considered that the composition sample prepared in Example 11 does not have synergistic penetration effect;

[0097] In summary, by comparing the 3h permeation rate with the 24h permeation rate, it can be seen that when the content of ergothioneine in the composition is 0.1wt%, and the content of polyglyceryl-10 laurate is 0.1-2wt%, i.e. the ratio of ergothioneine to polyglyceryl-10 laurate in the composition is 1:1-20, the composition has synergistic penetration effect. When the content of polyglyceryl-10 laurate in the composition is 0.03wt% and 3wt%, i.e. the ratio of ergothioneine to polyglyceryl-10 laurate is 10:3 or 1:30, the composition does not have penetration effect.

[0098] The 3h permeation rate of the composition sample prepared in Example 13 is 19.65%, which is greater than the 10.64% permeation rate of the composition sample prepared in Example 12 at 3h, and the 24h permeation rate of the composition sample prepared in Example 13 is 72.68%, which is greater than the 45.36% permeation rate of the composition sample prepared in Example 12 at 24h, and it is considered that the composition sample prepared in Example 13 has synergistic penetration effect; it can be seen that when the content of ergothioneine in the composition is 0.01wt%, and the content of polyglyceryl-10 laurate is 0.25wt%, i.e. the ratio of ergothioneine to polyglyceryl-10 laurate is 1:25, the composition has synergistic penetration effect.

[0099] The 3h permeation rate of the composition sample prepared in Example 15 is 20.80%, which is greater than the 11.41% permeation rate of the composition sample prepared in Example 14 at 3h, and the 24h permeation rate of the composition sample prepared in Example 15 is 66.30%, which is greater than the 59.70% permeation rate of the composition sample prepared in Example 14 at 24h, and it is considered that the composition sample prepared in Example 15 has synergistic penetration effect; it can be seen that when the content of ergothioneine in the composition is 0.001wt%, and the content of polyglyceryl-10 laurate is 0.01wt%, i.e. the ratio of ergothioneine to polyglyceryl-10 laurate is 1:10, the composition has synergistic penetration effect.

[0100] Test Example 2: Stability Test

[0101] Test sample: aqueous solution of ergothioneine and polyglyceryl-10 laurate with different proportions prepared in Examples 1-15

[0102] Stability evaluation method: place the sample in a constant temperature box at room temperature (25°C) and high temperature (48°C) respectively, and observe whether the sample has color change, crystal or particle precipitation, odor change problem at 1 week, 4 weeks respectively. After each time point, the sample is restored to room temperature, and then observed.

[0103] No color change, represented by o;

[0104] Color change or other changes, represented by x.

[0105] Odor scale: 0 for no change in odor (acceptable);

[0106] 1-2 for slight unpleasant odor (just acceptable);

[0107] 3-4 for presence of sulfur-like odor (unacceptable);

[0108] 5 for presence of strong sulfur-like odor, very unpleasant odor (unacceptable).

[0109] Table 3 Stability of ergothioneine and polyglyceryl-10 laurate in different proportions of aqueous solution

[0110]

[0111] After 1 week of stability test, the color of the compositions prepared in Examples 1-15 did not change, and no crystals or particles were precipitated, and the odor did not change at room temperature. The composition prepared in Example 1-11 had a slight unpleasant odor at high temperature. The compositions prepared in Examples 12-15 had no change in odor at room temperature and high temperature.

[0112] In summary, after 1 week of stability test, when the content of ergothioneine in the composition was 1wt%, 0.1wt%, a slight unpleasant odor appeared at high temperature, and it was still acceptable at room temperature. When the content of ergothioneine in the composition was 0.01wt%, 0.001wt%, there was no change in odor at room temperature and high temperature.

[0113] After 4 weeks of stability test, the color of the compositions prepared in Examples 1-15 did not change, and no crystals or particles were precipitated. The composition prepared in Example 1-11 had a slight unpleasant odor at room temperature and high temperature. The compositions prepared in Examples 12-15 had no change in odor at room temperature and high temperature.

[0114] In summary, after 4 weeks of stability test, when the content of ergothioneine in the composition was 1wt%, 0.1wt%, a slight unpleasant odor still appeared at room temperature and high temperature compared to the stability test results after 1 week. When the content of ergothioneine in the composition was 0.01wt%, 0.001wt%, there was no change in odor at room temperature and high temperature compared to the stability test results after 1 week.

[0115] Comparative Examples 1-5: Preparation of aqueous solutions of ergothioneine and different penetration enhancers

[0116] Sample preparation: Ergothioneine and different penetration enhancers were weighed according to the content shown in Table 4, deionized water was added to make up 100 parts by mass, and then stirred at room temperature until a transparent liquid was obtained, and then used.

[0117] Table 4. Content of each component in aqueous solutions of ergothioneine and different penetration enhancers.

[0118]

[0119] Note: The weight ratio of lauryl azone to RH40 in the table is 3:7.

[0120] Test Example 3: Permeability Test

[0121] Test samples: Aqueous solutions of ergothioneine prepared in Comparative Examples 1-5 with different penetration enhancers

[0122] Test steps:

[0123] 1) Remove the frozen pig skin, thaw it, and then use scissors to remove the subcutaneous fat and connective tissue.

[0124] 2) Soak the pigskin in 0.9wt% saline solution for 1 hour.

[0125] 3) Open as follows Figure 1 The vertical transdermal diffusion instrument shown is heated in a water bath (constant temperature 32℃).

[0126] 4) Place a magnetic stir bar in the receiving pool, then fix the cut pigskin between the supply and receiving pools, with the epidermis facing upwards and the dermis in contact with the receiving pool. Add physiological saline into the receiving pool through the sampling tube as the receiving solution, and remove any air bubbles. Cover the supply pool with a coverslip, place it in a transdermal diffusion apparatus, and equilibrate for 1 hour at a constant temperature (32℃) with stirring (440 r / min).

[0127] 5) Add each test sample into the supply pool, i.e., where the raw drug is located, in contact with the pigskin, and stir at a constant temperature (32℃) (440r / min). After 3h and 24h, take the receiving liquid from the sampling tube for testing.

[0128] 6) The ergothionein content in the receiving liquid was determined using high performance liquid chromatography.

[0129] a. Chromatographic conditions: The chromatographic column was an Agilent HILIC Plus column (4.6 mm * 100 mm, 3.5 μm), the mobile phase was 0.1% formic acid acetonitrile solution - 0.1% formic acid aqueous solution (80:20 v / v), the flow rate was 1 mL / min, the column temperature was 30 ℃, the injection volume was 10 μL, and the detection wavelength was 262 nm.

[0130] b. Preparation of ergothioneine standard stock solution (0.5 g / L): Weigh 50 mg of ergothioneine standard (accurate to 0.01 mg), dissolve in water and dilute to 100 mL. Store at 4 °C.

[0131] c. Ergothioneine standard working solution preparation: accurately pipette a certain amount of ergothioneine standard stock solution, and dilute it with 0.1% formic acid methanol solution to obtain a series of standard working solutions (mass concentrations are 5 μg / mL, 10 μg / mL, 25 μg / mL, 50 μg / mL, 100 μg / mL).

[0132] d. Standard curve drawing: according to the above chromatographic conditions, the standard working solution is determined, and the mass concentration of the standard working solution is taken as the abscissa and the peak area is taken as the ordinate to draw the standard curve.

[0133] e. Sample treatment and determination: weigh 0.1-0.5 g (accurate to 0.1 mg) of sample in a 10 mL stoppered colorimetric tube, add 8 mL of 0.1% formic acid methanol solution, and ultrasonic extraction for 20 min. Use 0.1% formic acid methanol solution to constant volume, shake well, take part of the liquid through 0.22 μm filter membrane, and determine according to the above chromatographic conditions. Quantify the sample on the standard curve. The response value of the sample solution to be tested should be within the linear range of the standard curve.

[0134] 7) Calculate the cumulative permeation rate

[0135] EGT permeation rate = (m 取样量 ×C EGT ) / (m 上样量 × CEGT总 )×100%

[0136] Table 5 Ergothioneine and water solution of different penetration enhancers

[0137]

[0138]

[0139] In this application, when the sample of the composition under investigation has a permeation rate of more than 50% in 24h, it is considered to have penetration enhancement effect.

[0140] Therefore, the sample of the composition prepared in Comparative Examples 1-3 has a permeation rate of more than 50% in 24h, that is, the sample of the composition prepared in Comparative Examples 1-3 has penetration enhancement effect.

[0141] The sample of the composition prepared in Comparative Examples 4 and 5 has no penetration enhancement effect; according to the permeability data of the sample of the composition prepared in Example 13 in Table 2, that is, the water solution of the combination of ergothioneine and polyglycerol-10 laurate has the highest permeation rate of 72.68%, and the highest penetration enhancement efficiency.

[0142] The epidermis of human skin is composed of a large number of lipid bilayers, so we generally choose raw materials with lipophilic and hydrophilic structures or polyols as penetration enhancers to help the penetration of active substances into the skin. The commonly used NeosolueTM -Aqulio (bis-diethoxydiglycol cyclohexane 1,4-dicarboxylate), RH40 is such a raw material that is both lipophilic and hydrophilic; and laurocapram, as one of the most commonly used penetration enhancers in the pharmaceutical field, helps the drug to be absorbed through the skin; in addition, polyols such as 1,2-pentanediol have good skin affinity and can also be used as a penetration enhancer to help the active substance penetrate. Polyglyceryl-10 laurate is commonly used as an emulsifier in the formulation of polyglyceryl-10 laurate. It is accidentally found that the penetration enhancement of polyglyceryl-10 laurate is better than that of the commonly used penetration enhancers listed above under the same proportion.

[0143] Polyglyceryl-10 myristate and inulin lauryl carbamate are emulsifiers, but unlike ergothioneine, polyglyceryl-10 myristate and inulin lauryl carbamate (Comparative Examples 4 and 5) have no penetration enhancement effect under the corresponding proportion, while polyglyceryl-10 laurate does.

[0144] Test Example 4: Stability test

[0145] Test sample: aqueous solutions of ergothioneine prepared in Comparative Examples 1-5 and different penetration enhancers

[0146] Stability evaluation method: place the samples in constant temperature ovens at room temperature (25°C) and high temperature (48°C) respectively, and observe the samples at 1 week and 4 weeks respectively. After each time point, the samples are restored to room temperature, and it is observed whether the samples have color change, crystal or particle precipitation, or odor change.

[0147] No color change, represented by ○;

[0148] Color change or other changes, represented by ×.

[0149] Table 6 Stability of aqueous solutions of ergothioneine and different penetration enhancers

[0150]

[0151]

[0152] After 1 week of stability, the color of the compositions prepared in Comparative Examples 1-5 had no change, and no crystals or particles were precipitated.

[0153] After 4 weeks of stability, the color of the compositions prepared in Comparative Examples 1-5 had no change, and no crystals or particles were precipitated.

[0154] In summary, after 1-4 weeks of stability test, when the content of ergothioneine in the composition is 0.01wt%, there is no color change or crystal precipitation at room temperature and high temperature.

[0155] Application Example

[0156] The compositions of polyglyceryl-10 laurate of Examples 1-15 and ergothioneine are used for the preparation of skin external agents. The skin external agents are preferably cosmetic compositions such as cosmetic water, serum, cream, pack, jelly, cleansing, shampoo, bath, make-up removal, etc. The compositions of polyglyceryl-10 laurate of Examples 1-15 are used in the skin external agents in a weight percentage of 0.01% to 20% (w / w). Preferably, the weight percentage is 0.01% to 10% (w / w). More preferably, the weight percentage is 0.01% to 5% (w / w).

[0157] The following are specific application examples of the compositions of polyglyceryl-10 laurate of Examples 1-15 in skin external agents, and the formulations and preparation methods of these agents. In the following tables, "-" indicates no addition.

[0158] Application Example 1: Preparation of a cream

[0159]

[0160]

[0161] Application Example 2: Preparation of an emulsion

[0162]

[0163] Application Example 3: Preparation of a jelly

[0164]

[0165] Application Example 4: Preparation of a cosmetic water

[0166]

[0167] Application Example 5: Preparation of a serum

[0168]

[0169] Application Example 6: Preparation of a pack

[0170]

[0171]

[0172] Application Example 7: Preparation of an eye cream

[0173]

[0174] Application Example 8: Preparation of an aerosol (cleansing foam)

[0175]

[0176]

[0177] Application Example 9: Preparation of a spray

[0178]

[0179] Application Example 10: Preparation of a shower gel

[0180]

[0181]

[0182] Application Example 11: Preparation of a facial cleanser

[0183]

[0184] Application Example 12: Preparation of a serum

[0185]

[0186]

[0187] In all of the above examples of specific applications of the skin external preparations, the polyglyceryl-10 laurate composition of Examples 1 to 15 can be used in the formulations either partially or wholly in place of deionized water or directly as the formulation.

Claims

1. A penetration enhancing composition based on polyglyceryl- 10 laurate consisting of ergothioneine, polyglyceryl- 10 laurate and water, wherein, The content of the polyglyceryl-10 laurate is 0.01wt%-2wt%, the weight ratio of the ergothioneine and the polyglyceryl-10 laurate is 2:1 to 1:25, the penetration rate of the composition after 24h vertical transdermal diffusion is ≥50%, the content of the ergothioneine is 0.001wt%-1wt%.

2. The composition of claim 1, wherein The content of the polyglyceryl-10 laurate is 0.01wt%-0.25wt%, the content of the ergothioneine is 0.001wt%-0.01wt%.

3. The composition of claim 1, wherein The weight ratio of the ergothioneine and the polyglyceryl-10 laurate is 1:1 to 1:

25.

4. The composition of claim 1, wherein The penetration rate of the composition after 24h vertical transdermal diffusion is 54.28-76.11%.

5. Use of polyglyceryl-10 laurate to promote the penetration of ergothioneine, characterized in that, The weight ratio of the ergothioneine and the polyglyceryl-10 laurate is 2:1 to 1:25, the content of the polyglyceryl-10 laurate is 0.01wt%-2wt%, the content of the ergothioneine is 0.001wt%-1wt%.

6. Use according to claim 5, characterized in that, The weight ratio of the ergothioneine and the polyglyceryl-10 laurate is 1:1 to 1:

25.

7. The use of the composition of claim 1 in a skin external preparation for non-therapeutic purposes.

8. The use of claim 7, wherein the content of the composition in the skin external preparation for non-therapeutic purposes is 0.01wt%-20wt%.

9. Use according to claim 8, wherein, The skin external preparation for non-therapeutic purposes is selected from the group consisting of cream, emulsion, essence, jelly, lotion, mask, cleanser, bath agent, make-up remover, hair washing and conditioning agent, oil agent, floral water.

Images