Water-soluble liquid preparation with high content of pedunculoside as well as preparation method and application of water-soluble liquid preparation

Abstract

The invention relates to a water-soluble liquid preparation with high content of pedunculoside as well as a preparation method and application of the water-soluble liquid preparation, and the liquid preparation is prepared from the following components in parts by weight: 2-3 parts of pedunculoside; 2 to 3 parts of lysine glutamate; 0.1 to 0.5 part of vegetable oil; 0.2 to 0.6 part of an emulsifier; 0.1 to 0.5 part of a co-emulsifier; 3-6 parts of deionized water; wherein the emulsifying agent is prepared from sorbitan olivate and sodium stearoyl glutamate, and the ratio of parts by weight of the sorbitan olivate to the sodium stearoyl glutamate ranges from (2 to 1) to (4 to 1). The water-soluble liquid preparation of pedunculoside is high in content of effective components, good in water solubility and high in stability, and compared with single pedunculoside, the water-soluble liquid preparation of pedunculoside has more remarkable skin soothing and oil control effects.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, and more specifically, relates to a water-soluble liquid preparation with high content of lecithin, its preparation method, and its application. Background Technology

[0002] Pedunculoside, also known as peony glycoside (PE), is a pentacyclic triterpenoid saponin compound with anti-tumor and antioxidant effects.

[0003] The aglycone of *Ilex longiflorum* glycoside has a triterpenoid skeleton and methyl / methylene substituents. The core of the triterpenoid skeleton consists of multiple nonpolar carbon rings (such as a cyclopentane-polyhydrophenanthrene structure), rich in hydrophobic C-H bonds, making it poorly soluble in water. Furthermore, the methyl and methylene groups on the aglycone further increase the hydrophobicity. This hydrophobicity not only results in poor water solubility, low bioavailability, and limited efficacy and uses, but also significantly restricts the expansion of its application scenarios.

[0004] Chinese patent application CN106074394A discloses a method for preparing a pharmaceutical composition to improve the dissolution rate of holly glycosides in poorly soluble drugs. This method utilizes solid dispersion technology, micronization technology, and pharmaceutically acceptable diluents or carriers to prepare a solid dosage form containing holly glycosides. Although the pharmaceutical composition prepared by this patent application can release holly glycosides, the content of holly glycosides in the pharmaceutical composition system is very low (approximately 6%), and the water solubility of the released holly glycosides is not fundamentally improved. Therefore, this patent application does not solve the technical problem of the water solubility of holly glycosides.

[0005] Chinese patent application CN119367272A discloses a thermosensitive injectable hydrogel containing holmite glycosides, its preparation method, and its application. The prepared microsphere-gel co-delivery formulation PE-M / Ge l exhibits good injectability and good thermosensitive gelation properties in vitro. The microspheres are uniformly distributed within the gel, making its release curve closer to zero-order release, and its dual controlled-release system enables more stable controlled-release of the drug. However, in this patent application, the drug loading of holmite glycosides is only 0.50-0.72% of the hydrogel system.

[0006] To address the current technical problems of poor water solubility, low bioavailability, limited efficacy, and narrow application scenarios of idemosiderin, this invention proposes a water-soluble liquid formulation with high content of idemosiderin, along with its preparation method and applications. Summary of the Invention

[0007] The main objective of this invention is to provide a water-soluble liquid preparation with high content of holly glycoside, as well as its preparation method and application, in order to solve at least one of the technical problems of holly glycoside in the prior art, such as poor water solubility, low bioavailability, limited efficacy and uses, and narrow application scenarios.

[0008] To achieve the above objectives, in a first aspect, the present invention provides a water-soluble liquid formulation with a high content of ilex glycosides, wherein the liquid formulation comprises the following components in parts by weight: 2-3 parts of long-stemmed holly glycosides; 2-3 parts of lysine glutamate; 0.1-0.5 parts vegetable oil; Emulsifier 0.2-0.6 parts; 0.1-0.5 parts of co-emulsifier; 3-6 parts deionized water; The weight ratio of the long-stemmed holly glycoside to lysine glutamate ranges from 1:1 to 1:1.5.

[0009] Preferably, the emulsifier is composed of sorbitan olive oil ester and sodium stearoyl glutamate, wherein the weight ratio of sorbitan olive oil ester to sodium stearoyl glutamate ranges from 2:1 to 4:1.

[0010] Preferably, the weight ratio of sorbitan olive oil ester to sodium stearoyl glutamate is 3:1.

[0011] Preferably, the vegetable oil is selected from one or more of the following: prickly pear fruit oil, peony seed oil, and peony seed oil.

[0012] Preferably, the co-emulsifier is selected from one or more combinations of pentylene glycol, glycerol, stearyl alcohol, butylene glycol, and polyethylene glycol.

[0013] Secondly, the present invention provides a method for preparing a water-soluble liquid formulation with a high content of holmyl glycoside, the preparation method comprising the following steps: Step 1: Mix lysine glutamate, holly glycoside and deionized water to obtain holly glycoside ionic liquid; Step 2: Mix the vegetable oil, emulsifier, and co-emulsifier to obtain the first emulsion; Step 3: Mix the long-stemmed holly glycoside ionic liquid with the first emulsion and stir at 40℃-50℃ to obtain the water-soluble liquid preparation with high content of long-stemmed holly glycoside.

[0014] Preferably, in step 1, lysine glutamate, ilexobin, and deionized water are mixed at 50°C-70°C to obtain ilexobin ionic liquid. More preferably, the mixture is mixed at 51°C-69°C; more preferably, at 52°C-68°C; more preferably, at 53°C-67°C; more preferably, at 54°C-66°C; more preferably, at 55°C-65°C; more preferably, at 56°C-64°C; more preferably, at 57°C-63°C; more preferably, at 58°C-62°C; more preferably, at 59°C-61°C; and more preferably, at 60°C.

[0015] Preferably, in step 1, the mixing is performed under ultrasonic conditions for 20-40 minutes to obtain a long-stemmed holly glycoside ionic liquid. More preferably, the ultrasonic duration is 21-39 minutes; more preferably, 22-38 minutes; more preferably, 23-37 minutes; more preferably, 24-36 minutes; more preferably, 25-35 minutes; more preferably, 26-34 minutes; more preferably, 27-33 minutes; more preferably, 28-32 minutes; more preferably, 29-31 minutes; and more preferably, 30 minutes.

[0016] Preferably, in step 3, the stirring temperature is 40℃-50℃. More preferably, it is 41℃-49℃; even more preferably, it is 42℃-48℃; even more preferably, it is 43℃-47℃; even more preferably, it is 44℃-46℃; even more preferably, it is 45℃. The stirring time is 5min-10min, more preferably, it is 7min.

[0017] Thirdly, this invention proposes the application of a water-soluble liquid preparation with a high content of holly glycosides in cosmetics for skin soothing and / or oil control effects. The water-soluble liquid preparation with a high content of holly glycosides can be used as an IL-1α inflammatory factor inhibitor and an SZ95 sebaceous cell sebum secretion inhibitor.

[0018] Preferably, the skin-soothing and / or oil-controlling effects of the water-soluble liquid formulation with high content of holly glycoside are superior to those of holly glycoside monomer.

[0019] Compared with the prior art, the present invention has the following beneficial effects: 1. This invention provides a water-soluble liquid formulation with high content of lysine glycosides, its preparation method, and its application. Lysine glutamate is dissolved in deionized water, wherein the -NH2 group in the lysine glutamate is protonated to obtain a lysine glutamate cation (containing -NH3). + The ester group of leucine glycoside is deprotonated to obtain the leucine glycoside anion (containing -COO). - The resulting long-stem holly anion (containing -COO) - ), lysine glutamate cation (containing -NH3) + The two can interact through charge, allowing ilex glycoside and lysine glutamate to form ilex glycoside ionic liquid in aqueous solution. This ionic liquid system gives ilex glycoside good water solubility.

[0020] 2. This invention provides a water-soluble liquid formulation with high content of holly glycoside, as well as its preparation method and application. The preparation method involves adding a first emulsion formed by mixing vegetable oil, emulsifier, and co-emulsifier. The first emulsion, when mixed with the ionic liquid, further enhances the water solubility of the holly glycoside ionic liquid. The vegetable oil in the first emulsion encapsulates the holly glycoside, while the compounded emulsifier and co-emulsifier provide better emulsification and dispersion, thereby improving the stability of the formulation. This results in a water-soluble liquid formulation with high content of holly glycoside exhibiting good water solubility and high stability.

[0021] 3. This invention provides a water-soluble liquid formulation with a high content of holmite glycoside, its preparation method, and its applications. The high-content, water-soluble holmite glycoside liquid formulation exhibits significant soothing effects in an in vitro 3D skin soothing model and can be used as an inhibitor of the inflammatory factor IL-1α. It also demonstrates significant oil-controlling effects in an in vitro SZ95 sebaceous gland cell sebum secretion model and can be used as an inhibitor of sebum secretion in SZ95 sebaceous cells. These soothing and oil-controlling effects are superior to those of holmite glycoside monomer. Therefore, this water-soluble liquid formulation with a high content of holmite glycoside can be widely used in functional cosmetic emulsions, aqueous solutions, sprays, and color cosmetics. Attached Figure Description

[0022] Figure 1 This is a schematic diagram of a method for preparing a water-soluble liquid formulation with high content of lecithin according to one embodiment of the present invention; Figure 2 The HPLC chromatograms are of ilex glycoside (standard) and HPLC chromatograms of the water-soluble liquid formulation of high-content ilex glycoside in Example 1 of this invention. Figure 3 The images show fluorescence microscopy staining of lipid secretion in SZ95 cells, as well as the oil-controlling effects of Ilex longiflora glycosides, according to Examples 1, 4, 5, and Comparative Example 7 of the present invention. Detailed Implementation

[0023] The various aspects of the present invention will be further described in detail below.

[0024] Unless otherwise defined or stated, all technical and scientific terms used herein have the same meaning as are familiar to a user skilled in the art. Furthermore, any methods and materials similar to or equivalent to those described herein may be applied to the methods of this invention.

[0025] Sources of relevant raw materials and materials: The holly glycosides (98% by weight) and the plant oil are both sourced from Yunnan Specialty Plant Extraction Laboratory Co., Ltd. Lysine glutamate, sorbitan olive oil ester, sodium stearoyl glutamate, stearyl alcohol, pentylene glycol, polyethylene glycol, etc. are all analytical grade and are sourced from Shanghai Yuanye Biotechnology Co., Ltd. Related instrument models and sources: High performance liquid chromatography: Agilent 1260 Infinity II HPLC-DAD; Ultrasonic: KQ-800DE CNC ultrasonic cleaner; The 3D skin is an Episkin large-pore skin model, purchased from Shanghai Sianfuno Biotechnology Co., Ltd. SZ95 sebaceous gland cells were derived from the Shanghai Cell Bank of the Chinese Academy of Sciences; The culture medium was purchased from Beijing Solarbio Science & Technology Co., Ltd.

[0026] In a first aspect, the present invention provides a water-soluble liquid formulation with a high content of holmyl glycoside, wherein the liquid formulation comprises the following components by weight: 2-3 parts of long-stemmed holly glycosides; 2-3 parts of lysine glutamate; 0.1-0.5 parts vegetable oil; Emulsifier 0.2-0.6 parts; 0.1-0.5 parts of co-emulsifier; 3-6 parts deionized water; The weight ratio of the long-stemmed holly glycoside to lysine glutamate ranges from 1:1 to 1:1.5.

[0027] See appendix Figure 1 Secondly, the present invention provides a method for preparing a water-soluble liquid formulation with a high content of lecithin, the preparation method comprising the following steps: Step 1: Mix lysine glutamate, holly glycoside and deionized water to obtain holly glycoside ionic liquid; Step 2: Mix the vegetable oil, emulsifier, and co-emulsifier to obtain the first emulsion; Step 3: Mix the ilicicolin H ionic liquid with the first emulsion and stir at 40°C - 50°C to obtain the water-soluble liquid preparation of ilicicolin H with high content.

[0028] Example 1

[0029] This example provides a preparation method for a water-soluble liquid preparation of ilicicolin H with high content. The preparation method includes the following steps: Step 1: By weight, mix 2 parts of ilicicolin H with a mass percentage content of 98%, 2.5 parts of lysine glutamate, and 4 parts of deionized water, and ultrasonicate at 60°C for 30 min to obtain the ilicicolin H ionic liquid. Step 2: Mix 0.3 parts of prickly pear seed oil, 0.4 parts of emulsifier, and 0.3 parts of pentylene glycol to obtain the first emulsion. The weight ratio of sorbitan oleate to sodium stearoyl glutamate in the emulsifier is 3:1. Step 3: Mix the ilicicolin H ionic liquid with the first emulsion and stir at 45°C for 7 min to obtain the water-soluble liquid preparation of ilicicolin H with high content.

[0030] Example 2

[0031] This example provides a preparation method for a water-soluble liquid preparation of ilicicolin H with high content. The preparation method includes the following steps: Step 1: By weight, mix 2 parts of ilicicolin H with a mass percentage content of 98%, 3 parts of lysine glutamate, and 6 parts of deionized water, and ultrasonicate at 65°C for 20 min to obtain the ilicicolin H ionic liquid. Step 2: Mix 0.1 parts of prickly pear seed oil, 0.2 parts of emulsifier, and 0.5 parts of stearyl alcohol to obtain the first emulsion. The weight ratio of sorbitan oleate to sodium stearoyl glutamate in the emulsifier is 2:1. Step 3: Mix the ilicicolin H ionic liquid with the first emulsion and stir at 50°C for 5 min to obtain the water-soluble liquid preparation of ilicicolin H with high content.

[0032] Example 3

[0033] This example provides a preparation method for a water-soluble liquid preparation of ilicicolin H with high content. The preparation method includes the following steps: Step 1: By weight, mix 2 parts of ilicicolin H with a mass percentage content of 98%, 2 parts of lysine glutamate, and 4 parts of deionized water, and ultrasonicate at 50°C for 40 min to obtain the ilicicolin H ionic liquid. Step 2: Mix 0.5 parts of sea buckthorn seed oil, 0.6 parts of emulsifier, and 0.1 parts of polyethylene glycol to obtain the first emulsion. The weight ratio of sorbitan olive oil ester to sodium stearoyl glutamate in the emulsifier is 4:1. Step 3: Mix the long-stemmed holly glycoside ionic liquid with the first emulsion and stir at 40°C for 10 minutes to obtain the water-soluble liquid preparation with high content of long-stemmed holly glycoside.

[0034] Example 4

[0035] This embodiment provides a liquid preparation of long-stemmed holly glycoside, which differs from Example 1 only in that the holly fruit oil is replaced with an equal amount of peony seed oil, while the other ingredients and proportions remain unchanged.

[0036] The preparation method is the same as in Example 1.

[0037] Example 5

[0038] This embodiment provides a liquid preparation of long-stemmed holly glycosides, which differs from Example 1 only in that the holly fruit oil is replaced with an equal amount of peony seed oil, while the other ingredients and proportions remain unchanged.

[0039] The preparation method is the same as in Example 1.

[0040] Comparative Example 1 The difference between Comparative Example 1 and Example 1 is that in step 1, the 2 parts of 98% long-stemmed holly glycosides remain unchanged, the weight of lysine glutamate is reduced to 1.8 parts, and the reduced amount of lysine glutamate is added to the other ingredients proportionally.

[0041] The preparation method is the same as in Example 1.

[0042] Comparative Example 2 The difference between Comparative Example 2 and Example 1 is that in step 1, the weight percentage of 98% holly glycoside is reduced to 1.56 parts, while the weight percentage of lysine glutamate remains unchanged at 2.5 parts. The reduced amount of holly glycoside is added to the other ingredients proportionally.

[0043] The preparation method is the same as in Example 1.

[0044] Comparative Example 3 The difference between Comparative Example 3 and Example 1 is that in step 2, the weight ratio of sorbitan olive oil ester to sodium stearoyl glutamate in the emulsifier is 1:1, while the other components and their proportions remain unchanged.

[0045] The preparation method is the same as in Example 1.

[0046] Comparative Example 4 The difference between Comparative Example 4 and Example 1 is that in step 2, the weight ratio of sorbitan olive oil ester to sodium stearoyl glutamate in the emulsifier is 5:1, while the other components and their proportions remain unchanged.

[0047] The preparation method is the same as in Example 1.

[0048] Comparative Example 5 The difference between Comparative Example 5 and Example 1 is that in step 1, 2 parts by weight of 98% ileoside, 2.5 parts by weight of lysine glutamate, and 4 parts by weight of deionized water remain unchanged. In step 2, the total weight of sorbitan olive oil ester and sodium stearoyl glutamate in the emulsifier is reduced to 0.1 parts, and the reduced amount of emulsifier is added to the other ingredients proportionally.

[0049] The preparation method is the same as in Example 1.

[0050] Comparative Example 6 The difference between Comparative Example 6 and Example 1 is that in step 1, the 2 parts by weight of 98% ileoside, 2.5 parts by weight of lysine glutamate, and 4 parts by weight of deionized water remain unchanged. In step 2, the total weight of sorbitan olive oil ester and sodium stearoyl glutamate in the emulsifier is increased to 0.7 parts, and the increased part of the emulsifier is proportionally deducted from the other ingredients.

[0051] The preparation method is the same as in Example 1.

[0052] Comparative Example 7 The difference between Comparative Example 7 and Example 1 is that in step 1, 2 parts by weight of 98% holly glycoside, 2.5 parts by weight of lysine glutamate, and 4 parts by weight of deionized water remain unchanged. In step 2, no vegetable oil is added, and the reduced weight percentage is proportionally allocated to the other ingredients.

[0053] The preparation method is the same as in Example 1.

[0054] Comparative Example 8 The difference between Comparative Example 8 and Example 1 is that in step 1, the two portions of 98% long-stemmed holly glycosides remain unchanged, lysine glutamate is not added, and the reduced weight portion is proportionally allocated to other ingredients.

[0055] The preparation method is the same as in Example 1.

[0056] Comparative Example 9 The difference between Comparative Example 9 and Example 1 lies in their preparation methods. The preparation method for Comparative Example 9 is as follows: By weight, 2 parts of 98% holly glycoside, 2.5 parts of lysine glutamate, 4 parts of deionized water, 0.3 parts of prickly pear oil, 0.4 parts of emulsifier (the emulsifier contains sorbitan olive oil ester and sodium stearoyl glutamate in a weight ratio of 3:1), and 0.3 parts of pentylene glycol are mixed and ultrasonically treated at 60°C for 30 min, followed by stirring at 45°C for 7 min to obtain the water-soluble liquid preparation with high holly glycoside content.

[0057] Test Example 1: Stability Study and Determination of Ilex longiflora glycoside Content Test sample: The appearance and content of high-content ilex glycosides in the water-soluble liquid preparations prepared in Examples 1-5 and the ilex glycoside preparations prepared in Comparative Examples 1-9 were tested on day 0 and at one month after being placed at room temperature.

[0058] Test method: The test samples prepared in Examples 1-5 and Comparative Examples 1-9 were diluted 10 times with deionized water, filtered, and the content of ide glycosides in them was determined by HPLC-DAD. The results are shown in Table 1.

[0059] Test conditions: Chromatographic column: ZORBAX SB-C18 column (4.6×250 mm; 5-Micron); mobile phase: water (A)-acetonitrile (B) (0-10 min: 10% B; 10-20 min: 10% B-40% B; 20-30 min: 40% B); detection wavelength: 210 nm; column temperature: 30℃; flow rate: 1 mL / min; injection volume: 10 µL.

[0060] HPLC chromatograms of lecithin (standard) and the water-soluble liquid formulation of high-content lecithin prepared in Example 1 are shown below. Figure 2 As shown.

[0061] Table 1. Appearance and percentage content of ide glycosides in the water-soluble liquid formulations prepared in Examples 1-5 and Comparative Examples 1-9, corresponding to day 0 and one month of storage at room temperature.

[0062] As shown in Table 1, among the water-soluble preparations of holly glycosides prepared in Examples 1-5 of this invention, the water-soluble preparations of holly glycosides prepared in Examples 1 and 3-5 have a holly glycoside content of over 20% by mass, and the water-soluble preparation of holly glycosides prepared in Example 2 has a holly glycoside content of 16.53% by mass. Furthermore, the water-soluble preparations of holly glycosides prepared in Examples 1-5, after being stored for one month, exhibit the same properties as when they were first prepared, all being clear, transparent, oily liquids. These clear, transparent, oily liquids are miscible with water in any proportion, exhibiting strong water solubility, and their aqueous solutions are clear and transparent. Therefore, the water-soluble preparations of holly glycosides prepared in Examples 1-5 of this invention possess excellent stability and water solubility, maintaining a clear, transparent, oily liquid state for at least one month, and with a holly glycoside content exceeding 16%.

[0063] Conversely, the formulations prepared in Comparative Examples 1-9, after being left at room temperature for one month, either exhibited slight white flocculent precipitation or a white precipitate. This indicates that the formulations prepared in Comparative Examples 1-9 have poor stability. Specifically, the formulations prepared in Comparative Examples 1-3, 6, 7, and 9 appeared as clear, transparent, oily liquids on day 0, but the appearance of slight white flocculent precipitation after one month at room temperature indicates poor stability; while the formulations prepared in Comparative Examples 4-5 and 8 appeared as milky white, turbid liquids on day 0. Therefore, it can be seen that adjusting the weight ratio of ilex glycoside to lysine glutamate outside the range of 1:1 to 1:1.5 in Comparative Examples 1-2 will affect the stability of the prepared formulation; adjusting the weight ratio of sorbitan olive oil ester to sodium stearoyl glutamate in the emulsifier outside the range of 2:1 to 4:1 in Comparative Examples 3-4 will adversely affect the stability of the prepared formulation and the water solubility of ilex glycoside; similarly, adjusting the weight of the emulsifier outside the range of 0.2-0.6 parts in Comparative Examples 5-6 will adversely affect the stability of the prepared formulation and the water solubility of ilex glycoside; and omitting vegetable oil or lysine glutamate or using the "one-pot method" in Comparative Examples 7-9 will adversely affect the stability of the prepared formulation and the water solubility of ilex glycoside.

[0064] Test Example 2: Efficacy Detection of In Vitro Reconstructed 3D Epidermal Model Test samples: the water-soluble liquid formulations of holly glycoside prepared in Examples 1, 4 and 5, and the formulation prepared in Comparative Example 7 with a mass percentage content of 98% holly glycoside.

[0065] Test method: A blank control group, an SLS (sodium dodecyl sulfate) induced stimulation group (negative control group), a positive control group (dexamethasone, dextromethorphan), and a sample group were set up (in the sample group, the final concentration of ilexobin was 1200 μg / mL, dissolved and added to the culture medium). All groups were incubated at 37°C in a 5% CO2 incubator for 2 h. Except for the blank control group, SLS was added to each well, and the cells were incubated for 24 h. The cell supernatant was collected, and the inhibitory effect of IL-1α was detected by ELISA.

[0066]

[0067] Table 2. IL-1α inhibition rate of the water-soluble liquid formulations of halomena glycoside prepared in Examples 1, 4, and 5, compared with that of the formulation prepared in Comparative Example 7, and the halomena glycoside content of 98%.

[0068] Table 2 shows that the water-soluble liquid formulations of holly glycoside prepared in Examples 1, 4, and 5 exhibited good inhibitory effects on IL-1α inflammatory factors. Specifically, the 70.63% IL-1α inflammatory factor inhibition rate of the water-soluble liquid formulation prepared in Example 1 was close to that of the positive control group (dexamethasone, dextromethorphan). The water-soluble liquid formulations of holly glycoside prepared in Examples 4 and 5 also showed higher IL-1α inhibition rates than the formulation prepared in Comparative Example 7 and the 98% holly glycoside content formulation, indicating that *Ilex chinensis* fruit oil, *Paeonia suffruticosa* seed oil, and *Paeonia lactiflora* seed oil can synergistically enhance the soothing efficacy of holly glycoside.

[0069] Test Example 3: Oil Control Efficacy Test Test samples: the water-soluble liquid formulations of holly glycoside prepared in Examples 1, 4 and 5, and the formulation prepared in Comparative Example 7 with a mass percentage content of 98% holly glycoside.

[0070] Test method: The lipid droplets were induced by 5α-dihydrotestosterone (DHT) and positively charged by isotretinoin. The oil-controlling effect was evaluated by detecting the inhibitory effect of the test sample on the secretion of SZ95 sebaceous gland cells.

[0071] Press 1×10 5 Cells were seeded into 24-well plates at a seeding density of cells / well and incubated overnight in an incubator (37°C, 5% CO2). When the cell deposition rate in the 24-well plates reached 80% to 90%, the cells were administered to the appropriate groups.

[0072] The control group consisted of a blank group (cultured in culture medium only, without any treatment), a negative control group (5α-dihydrotestosterone (DHT)), a positive control group (isotretinoin), and a sample group (all samples in the sample group were prepared with a final concentration of 120 μg / mL of ide, dissolved and then added to the culture medium). Each group was divided into three replicates, and the cells were incubated in an incubator (37℃, 5% CO2) for 24 h. The culture medium was discarded, and the cells were rinsed with PBS. After fixation, Nile red staining was added for 15 min, and after rinsing, the cells were photographed using a fluorescence microscope. Figure 3 As shown. Quantitative analysis of fluorescence intensity was performed using Image Pro Plus software.

[0073] Formula for calculating oil inhibition rate:

[0074] Table 3. Oil inhibition rate of the water-soluble liquid formulations of holly glycoside prepared in Examples 1, 4, and 5, compared with the formulation prepared in Comparative Example 7, and the holly glycoside content of 98%. Group Oil inhibition rate (%) Positive control group 63.21% Example 1 56.82% Example 4 53.57% Example 5 50.16% Comparative Example 7 40.23% Longstem holly glycoside 38.49% As shown in Table 3, the water-soluble liquid formulations of holly glycoside prepared in Examples 1, 4, and 5 exhibited good lipid inhibition effects, with lipid inhibition rates all exceeding 50%. Specifically, the water-soluble liquid formulation of holly glycoside prepared in Example 1 showed a lipid inhibition effect of 56.82%. In contrast, the formulation prepared in Comparative Example 7 and the holly glycoside with a content of 98% showed an lipid inhibition rate of approximately 40%. This indicates that *Ilex chinensis* fruit oil, *Paeonia suffruticosa* seed oil, and *Paeonia lactiflora* seed oil can synergistically enhance the lipid-controlling effect of holly glycoside.

[0075] Therefore, the water-soluble liquid formulation of *Ilex longiflorus* glycoside prepared in this invention has significant effects in inhibiting IL-1α inflammatory factors and inhibiting sebum secretion from SZ95 sebaceous gland cells. This formulation can act as an inhibitor of IL-1α inflammatory factors and an inhibitor of sebum secretion from SZ95 sebaceous gland cells, and its anti-inflammatory and oil-controlling effects are significantly better than those of the monomeric *Ilex longiflorus* glycoside. Furthermore, the addition of *Rhizoma Cirsium japonicum* oil, *Paeonia suffruticosa* seed oil, and *Paeonia suffruticosa* seed oil to the water-soluble formulation of *Ilex longiflorus* glycoside can synergistically enhance its effects, thereby improving its soothing and oil-controlling efficacy.

[0076] Based on this invention, those skilled in the art will understand that one aspect described herein can be implemented independently of any other aspect, and two or more of these aspects can be combined in various ways. For example, any number and aspects set forth herein can be used to implement the device and / or practice the method. Furthermore, this device and / or practice the method can be implemented using other structures and / or functionalities besides one or more of the aspects set forth herein.

[0077] It should be noted that the above embodiments can be freely combined as needed. The above description is only a preferred embodiment of the present invention. It should be pointed out that for users of ordinary skills in this field, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

[0078] All documents mentioned in this invention are incorporated herein by reference as if each document were individually incorporated by reference. Furthermore, it should be understood that after reading the foregoing description of this invention, users skilled in the art can make various alterations or modifications to this invention, and these equivalent forms also fall within the scope defined by the appended claims.

Claims

1. A water-soluble liquid preparation with a high content of lecithin, characterized in that, The liquid formulation comprises, by weight, the following components: 2-3 parts of long-stemmed holly glycosides; 2-3 parts of lysine glutamate; 0.1-0.5 parts vegetable oil; Emulsifier 0.2-0.6 parts; 0.1-0.5 parts of co-emulsifier; 3-6 parts deionized water; The weight ratio of the long-stemmed holly glycoside to lysine glutamate ranges from 1:1 to 1:1.

5.

2. The water-soluble liquid preparation with high content of holmyl glycoside according to claim 1, characterized in that, The emulsifier is composed of sorbitan olive oil ester and sodium stearoyl glutamate, wherein the weight ratio of sorbitan olive oil ester to sodium stearoyl glutamate ranges from 2:1 to 4:

1.

3. The water-soluble liquid preparation with high content of holmyl glycoside according to claim 1, characterized in that, The vegetable oil is selected from one or more of the following: prickly pear fruit oil, peony seed oil, and peony seed oil.

4. The water-soluble liquid preparation with high content of holmyl glycoside according to claim 1, characterized in that, The co-emulsifier is selected from one or more combinations of pentylene glycol, glycerol, stearyl alcohol, butylene glycol, and polyethylene glycol.

5. A method for preparing a water-soluble liquid formulation with high content of lecithin as described in any one of claims 1-4, characterized in that, The preparation method includes the following steps: Step 1: Mix lysine glutamate, holly glycoside and deionized water to obtain holly glycoside ionic liquid; Step 2: Mix the vegetable oil, emulsifier, and co-emulsifier to obtain the first emulsion; Step 3: Mix and stir the long-stemmed holly glycoside ionic liquid with the first emulsion to obtain the water-soluble liquid preparation with high content of long-stemmed holly glycoside.

6. The method for preparing the water-soluble liquid formulation with high content of lecithin according to claim 5, characterized in that, In step 1, lysine glutamate, leucocephalin, and deionized water are mixed at 50℃-70℃ to obtain leucocephalin ionic liquid.

7. The method for preparing the water-soluble liquid formulation with high content of lecithin according to claim 5, characterized in that, In step 1, the mixing is carried out under ultrasonic conditions for 20-40 minutes to obtain long-stemmed holly glycoside ionic liquid.

8. The method for preparing the water-soluble liquid formulation with high content of lecithin according to claim 5, characterized in that, In step 3, the stirring temperature is 40℃-50℃ and the stirring time is 5-10 minutes.

9. The use of a water-soluble liquid formulation of high content of holmyl glycoside as described in any one of claims 1-4 in cosmetics with skin-soothing and / or oil-controlling effects.

10. The application of the water-soluble liquid formulation with high content of holmyl glycoside according to claim 9 in cosmetics with skin-soothing and / or oil-controlling effects, characterized in that, The water-soluble liquid formulation with high content of Ilex chinensis glycoside has better skin-soothing and / or oil-controlling effects than the monomeric Ilex chinensis glycoside.

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