A liposome with lipase responsiveness, its preparation method and application and product

Abstract

The application discloses a bacteriostatic liposome with lipase responsiveness, a preparation method and application thereof and a product, and belongs to the technical field of cosmetics. The technical problems to be solved are that existing means for treating acne are faced with problems such as drug resistance, high skin irritation and the like. Meanwhile, bacteriostatic core materials such as paeonol have problems such as poor solubility, poor transdermal absorption, burst stimulation and the like. The technical solution is characterized by providing a bacteriostatic liposome with lipase responsiveness, wherein wall materials include soybean phosphatidylcholine, hydrogenated phosphatidylcholine, cholesterols, PEG-7 olive oil acid ester, glycerol stearate and ceramide NP; and internal core materials include at least one of paeonol, paeonol, honokiol, caprylyl glycine, glabridin, ascorbic acid tetraisopalmitate, tetrahydrocurcumin or pterostilbene. Through a specific preparation method, the release speed of the core materials is regulated by using the reaction between lipase and the wall materials, a slow-release and controlled-release effect is achieved, and the product is easy to be produced industrially.

Description

Technical Field

[0001] This invention relates to the field of cosmetic technology, specifically to a liposome with lipase responsiveness, its preparation method, applications, and products. Background Technology

[0002] Responsive delivery systems are intelligent, targeted drug delivery methods that can achieve sustained or controlled release of drugs in response to changes in the external environment. Enzymes play a triggering role in these systems. They possess unique properties, including substrate specificity and high selectivity under appropriate conditions. Because these processes are inherently biological and metabolic, the distribution of lipases on the skin surface can be utilized for controlled drug release.

[0003] Oily skin is a widespread skin phenotype, affecting over 40% of the adult population globally, and more than 80% in adolescents. Its characteristic features include excessive sebum secretion from the sebaceous glands, accompanied by enlarged pores, dull skin tone, and increased susceptibility to acne. Acne vulgaris, a chronic inflammatory skin disease, involves multiple interacting factors in its pathogenesis, including abnormal keratinization of the pilosebaceous duct, excessive proliferation of Propionibacterium acnes, hypersecretion of sebum, and dysregulation of the inflammatory response. Therefore, it has long troubled many people. According to a World Health Organization survey, over 1 billion people worldwide suffer from acne, with a prevalence rate as high as 85% among adolescents. While current treatments can alleviate acne, they still face challenges such as drug resistance and high skin irritation.

[0004] Lipase is an enzyme secreted by bacteria that effectively hydrolyzes fats into free fatty acids. Studies have shown that in the pathological microenvironment of acne, *Propionibacterium acnes* secretes large amounts of lipase, with concentrations reaching up to 200 U / mL in hair follicles. Excessive lipase levels can lead to localized skin inflammation, abnormal sebum secretion, or excessive sebum oxidation. Therefore, designing a lipase-responsive delivery vehicle based on the pathological microenvironment of acne will be a major breakthrough in drug delivery, enabling efficient and precise drug delivery while also achieving sustained-release and controlled-release effects.

[0005] Relevant patent documents retrieved:

[0006] The document, published in China (CN119454489A) on February 18, 2025, discloses a liposome precursor composition comprising 40-53% polyol, 35-45% panthenol, 10-15% lecithin, and 2-4% ceramide NP. This composition can improve the incompatibility of ceramides with common formulation systems and enhance the stability, water solubility, and permeability of the ceramide system on the skin surface.

[0007] Relevant non-patent literature retrieved:

[0008] The journal or book title is "Food Industry Technology," and the article title is "The Influence of Phytosterols and Cholesterol on Liposome Membrane Properties." This article discloses that cholesterol has been extensively studied as a membrane structure regulator. Phytosterols have a similar structure to cholesterol and can inhibit the intestinal absorption of cholesterol. Replacing cholesterol with phytosterols can regulate the structural properties of liposome membranes while avoiding the negative effects of cholesterol. The article also explores the effects of different sterols on liposome particle size, antioxidant properties, membrane micropolarity, and membrane fluidity. Summary of the Invention

[0009] The purpose of this invention is to provide:

[0010] A liposome with lipase responsiveness, its preparation method and application and products, and related technologies, to solve the technical problems of poor targeting and inability to fully achieve controlled release in existing liposome skin delivery, or a combination thereof.

[0011] Terminology Explanation:

[0012] Unless otherwise defined, all technical terms in this document have the same meanings as commonly understood by one of ordinary skill in the art to which the subject matter of the claims pertains. Unless otherwise stated, all patents, patent inventions, and publications cited in this document are incorporated herein by reference in their entirety. If multiple definitions exist for terms in this document, the definitions in this chapter shall prevail.

[0013] It should be understood that the above brief description and the following detailed description are exemplary and for illustrative purposes only, and do not limit the subject matter of the invention in any way. In this invention, the singular is used in conjunction with the plural unless otherwise specifically stated. It should also be noted that, unless otherwise stated, the use of “or” or “or” means “and / or”. Furthermore, the use of the term “comprising” and other forms such as “including,” “containing,” and “contains” are not limiting.

[0014] The definition of standard chemical terms can be found in the reference "Complete Guide to Cosmetic Chemistry and Process Technology (Volumes 1 & 2)," edited by Qiu Bingyi, published by China Light Industry Press.

[0015] Unless otherwise stated, conventional methods within the scope of the art, such as the MTT assay for cytotoxicity and ultraviolet spectrophotometry for release, shall be used.

[0016] Unless specifically defined herein, the use of all commercially available products herein employs standard techniques. For example, it may be carried out using the manufacturer's instructions for use with the kit, or in accordance with methods known in the art or the description of this invention. The techniques and methods described herein can generally be implemented according to conventional methods well known in the art, based on the descriptions in the various summary and more specific documents cited and discussed in this specification.

[0017] The terms "optional / arbitrary" or "optionally / arbitrarily" mean that an event or situation described below may or may not occur, including both the occurrence and non-occurrence of the event or situation. For example, "optionally added antioxidant" means "adding an antioxidant" or "not adding an antioxidant."

[0018] The term "lipase responsiveness" used in this article refers to the ability of the wall material of liposomes to undergo hydrolysis under the action of lipases, thereby changing the structure of the liposomes and releasing the core material.

[0019] As used in this article, "liposomes" refers to closed vesicles formed by a lipid bilayer that can encapsulate water-soluble or lipid-soluble drugs.

[0020] The term "tau phenol" used in this article refers to a phenolic compound extracted from plants of the Myrtaceae family, which has good antibacterial effects.

[0021] In a first aspect, the present invention provides: an antibacterial liposome with lipase responsiveness.

[0022] This includes: wall material, core material, lipid content, drug-lipid ratio, etc.

[0023] The wall material includes lipids.

[0024] The lipid is selected from at least three of the following: soybean phosphatidylcholine, hydrogenated phosphatidylcholine, ceramide NP, polyglycerol-4, polyglycerol-10, PEG-50 shea butter, 1,2-distearate-sn-glycerol-3-phosphate ethanolamine (DSPE), glyceryl stearate, lysophosphatidylcholine, PEG-7 olive oil ester, PEG-10 olive oil glyceride, PEG-7 glyceryl cocoate, and cholesterol.

[0025] The lipids are selected from at least six of the following: soybean phosphatidylcholine, hydrogenated phosphatidylcholine, ceramide NP, polyglycerol-4, polyglycerol-10, PEG-50 shea butter, DSPE, glyceryl stearate, lysophosphatidylcholine, PEG-7 olive oil ester, PEG-10 olive oil glyceride, PEG-7 glyceryl cocoate, and cholesterol.

[0026] The lipids are preferably soybean phosphatidylcholine, hydrogenated phosphatidylcholine, cholesterol, PEG-7 olive oil ester, glyceryl stearate, and ceramide NP.

[0027] The lipids are further preferably 8-16 parts of soybean phosphatidylcholine, 0.5-1.5 parts of hydrogenated phosphatidylcholine, 0.3-0.8 parts of cholesterol, 2-3 parts of PEG-7 olive oil ester, 2-3 parts of glyceryl stearate, and 2-3 parts of ceramide NP.

[0028] The lipids are more preferably 8-12 parts of soybean phosphatidylcholine, 0.5-1 part of hydrogenated phosphatidylcholine, 0.3-0.5 parts of cholesterol, 2-3 parts of PEG-7 olive oil ester, 2-3 parts of glyceryl stearate, and 2-3 parts of ceramide NP.

[0029] The core material includes medicinal materials selected from: tau phenol, paeonol, magnolol, capryloyl glycine, glycyrrhizin, ascorbate tetraisopalmitate, or pterostilbene.

[0030] The preferred medicinal materials are: tau tannin, paeonol, glycyrrhizin, and ascorbate tetraisopalmitate.

[0031] Among them, the medicinal materials are further selected as follows: tau phenol and paeonol.

[0032] Among them, the preferred medicinal material is tau phenol.

[0033] The lipid content is selected from 3% to 10%.

[0034] The preferred lipid content is 5%-8%.

[0035] The lipid content is further preferably 6%-7%.

[0036] The drug-to-lipid ratio is selected from 1:30 to 1:50.

[0037] The preferred drug-to-lipid ratio is 1:16-1:24.

[0038] Based on further solutions to the technical problems of the present invention, or simultaneous solutions to multiple technical problems, the preferred solution in the technical solution provided in the first aspect of the present invention includes:

[0039] The first preferred option comprises a wall material consisting of soybean phosphatidylcholine, hydrogenated phosphatidylcholine, cholesterol, PEG-7 olive oil ester, glyceryl stearate, and ceramide NP, with a core material consisting of tau phenol. This technical solution addresses the technical challenges of tau phenol, such as poor solubility, poor transdermal absorption, and burst release stimulation, while further improving lipase responsiveness and sustained-release effect.

[0040] The second preferred solution comprises a wall material consisting of soybean phosphatidylcholine, hydrogenated phosphatidylcholine, cholesterol, PEG-7 olive oil ester, glyceryl stearate, and ceramide NP; and a core material consisting of tau phenol, with a lipid content of 3-10% and a drug-to-lipid ratio of 1:10-1:3, forming lipase-responsive antibacterial liposomes. This solution, by addressing the aforementioned technical challenges, further achieves sustained-release, controlled-release, and precise release of the core material.

[0041] Secondly, the present invention provides a method for preparing antibacterial liposomes with lipase responsiveness, comprising the following steps:

[0042] S1: Weigh out the lipids and medicinal materials according to the above mass ratio, dissolve them in ethanol solution, and heat until transparent and clear.

[0043] S2: Take a round-bottom flask, add an appropriate amount of PBS buffer solution, and keep it warm.

[0044] S3: Slowly inject the ethanol solution into the PBS buffer while stirring slowly.

[0045] S4: Pour the above liquid into a high-pressure microjet device, and circulate it under pressure for 1-5 times to obtain the final product.

[0046] This includes: PBS buffer pH value, incubation temperature, stirring time, high-pressure microjet pressure, number of cycles, etc.

[0047] The pH value of the PBS buffer is selected from 6.5-7.2.

[0048] The preferred pH value of the PBS buffer is 6.8-7.0.

[0049] The pH value of the PBS buffer is further preferably 6.8.

[0050] The insulation temperature is selected from 30℃-50℃.

[0051] The preferred insulation temperature is 35℃-45℃.

[0052] The optimal insulation temperature is 40℃.

[0053] The stirring time is selected from 30-90 min.

[0054] The preferred stirring time is 50-70 minutes.

[0055] The stirring time is further preferably 60 minutes.

[0056] The pressure of the high-pressure microjet is selected from 15000Psi-30000Psi.

[0057] The pressure of the high-pressure microjet is preferably 15,000-20,000 Psi.

[0058] The pressure of the high-pressure microjet is further preferably 15000 Psi.

[0059] The number of homogenization cycles is selected from 1 to 5.

[0060] The preferred number of homogenization cycles is 2-4.

[0061] The homogenization cycle is further optimized to be 3 times.

[0062] Based on further solutions to the technical problems of the present invention, or simultaneous solutions to multiple technical problems, the preferred solution in the technical solution provided in the second aspect of the present invention includes:

[0063] The first preferred method involves using ethanol solution as the dissolving solvent, 6.5-7.2 PBS buffer, incubation at 30-50℃, stirring for 30-90 min, high-pressure microfluidic jet at 15000-30000 psi, and 5 cycles. This method addresses issues such as uneven dissolution and excessively large particle size during liposome preparation, further improving the stability and encapsulation efficiency of liposomes.

[0064] Thirdly, the present invention provides the application of the liposomes or liposomes prepared by the above-described preparation method in the preparation of cosmetics or pharmaceuticals.

[0065] The cosmetics include, but are not limited to: acne treatment lotion, acne treatment essence, lotion, and face cream; the medicines include, but are not limited to, gel and cream.

[0066] Fourthly, the present invention provides a product in which an appropriate amount of liposomes or liposomes prepared by a preparation method are added; the product is a cosmetic or a pharmaceutical.

[0067] The amount added is selected from 0.1% to 1%.

[0068] The preferred addition amount is 0.5%-0.9%.

[0069] The preferred addition amount is 0.7%-0.9%.

[0070] The phrase "the wall material is composed of three or more of a variety of materials" is summarized from the foregoing explanation and the corresponding wall material combinations in Examples 1-3, based on the common characteristic of "having lipase responsiveness." Therefore, those skilled in the art can reasonably infer that the subordinate concepts of this wall material characteristic, substantially equivalent technical means (such as replacing some lipid materials with similar functions), etc., should all fall within the scope of protection of this invention.

[0071] "The core material is composed of one or more of a variety of materials," which is summarized from the selection of core materials in Examples 1-3 (pursulphol, glycyrrhizin, pterostilbene, ascorbate tetraisopalmitate). Similarly, replacing them with the same type of antibacterial and anti-inflammatory components (such as eugenol, bisabolol, hesperidin, etc.) is also within the scope of protection of this invention.

[0072] The present invention has at least the following beneficial effects:

[0073] 1. Compared with existing technologies, this invention has advantages in core material stability, transdermal absorption, and irritation, and also exhibits significantly improved lipase responsiveness, providing effective support for controlled release. It offers better technical results.

[0074] According to experimental tests, the liposomes of the present invention, after being stored at 4℃ and 45℃ for 30 days, showed an average particle size change rate of less than 5% and an absolute change in zeta potential of less than 3mV, while existing ordinary liposomes showed a particle size change rate of more than 15% and an absolute change in zeta potential of more than 8mV under the same conditions; the survival rate of HaCaT cells and RAW264.7 cells was not less than 85%, while the cell survival rate of tau tannin suspension at the same concentration was less than 70%. Attached Figure Description

[0075] Figure 1 This is a statistical chart showing the toxic effects of the liposomes prepared in Example 1 of this invention on RAW 264.7 cells;

[0076] Figure 2 This is a statistical chart showing the toxic effects of liposomes prepared in Example 1 of this invention on HaCat cells;

[0077] Figure 3 This is a graph showing the cumulative permeation amount of the liposomes prepared in Example 1 of the present invention in an in vitro transdermal diffusion experiment;

[0078] Figure 4 The in vitro lipase response release curve of liposomes prepared in Example 1 of this invention;

[0079] Figure 5 The in vitro lipase response release curve of the liposomes prepared in Comparative Example 1 of this invention;

[0080] Figure 6The in vitro lipase response release curve of liposomes obtained in Example 3 of this invention;

[0081] Figure 7 The in vitro lipase response release curve of liposomes obtained in Example 4 of this invention;

[0082] Figure 8 The in vitro lipase response release curve of liposomes obtained in Example 5 of this invention;

[0083] Figure 9 This is a test of the in vitro lipase responsiveness of liposomes prepared in Example 1 and Comparative Examples 3-5 of the present invention.

[0084] Figure 10 This is a test diagram of the antibacterial effect of the liposomes prepared in Example 1 of the present invention against Propionibacterium acnes.

[0085] Figure 11 This is a test diagram of the antibacterial effect of the liposomes prepared in Example 1 of the present invention against Staphylococcus aureus.

[0086] Figure 12 This is an experimental diagram showing the effect of liposomes prepared in Example 1 of the present invention on lipid peroxidation;

[0087] Figure 13 The figure shows the stability analysis test results of the liposomes prepared in Example 1 of this invention.

[0088] Figure 14 This is a particle size distribution diagram of liposomes obtained in Example 1 of the present invention;

[0089] Figure 15 This is a graph showing the particle size variation of the liposomes prepared in Example 1 of the present invention under various storage conditions. Detailed Implementation

[0090] The following non-limiting embodiments are intended to enable those skilled in the art to gain a more comprehensive understanding of the present invention, but do not limit the invention in any way. The following content is merely an exemplary description of the scope of protection claimed by the present invention, and those skilled in the art can make various changes and modifications to the present invention based on the disclosed content, and such changes should also fall within the scope of protection claimed by the present invention.

[0091] The present invention will be further described below by way of specific embodiments. Unless otherwise specified, all instruments, devices, equipment, reagents, products, etc., used in the embodiments of the present invention are obtained through conventional commercial means.

[0092] Example 1

[0093] Preparation of lipophenol liposomes with lipase responsiveness

[0094] (1) Weigh out soybean phosphatidylcholine, hydrogenated phosphatidylcholine, cholesterol, PEG-7 olive oil ester, glyceryl stearate, ceramide NP and tau phenol and dissolve them in 20 mL of ethanol solution. Heat until the solution is clear and transparent. The specific dosage is shown in Table 1.

[0095] Table 1. Distribution ratio of each group in Example 1

[0096]

[0097] (2) Take a round-bottom flask, add the mixed solution obtained in step (1), and then add 30 mL of PBS buffer solution with pH=6.8, and keep warm to 40℃.

[0098] (3) Slowly inject 20 mL of ethanol solution into PBS buffer and stir slowly for 60 min.

[0099] (4) Pour the liquid obtained in step (3) into a high-pressure microfluidic device and homogenize it three times at a pressure of 15000psi to obtain the final product. The sample obtained in this embodiment is a pale yellow transparent liquid.

[0100] Example 2

[0101] Preparation of lipophenol liposomes with lipase responsiveness

[0102] (1) Weigh out soybean phosphatidylcholine, hydrogenated phosphatidylcholine, cholesterol, PEG-7 olive oil ester, glyceryl stearate, ceramide NP and tau phenol and dissolve them in ethanol solution. Heat until the solution is clear and transparent. The specific dosage is shown in Table 2.

[0103] Table 2. Distribution ratio of each group in Example 2

[0104]

[0105] (2) Take a round-bottom flask, add the mixed solution obtained in step (1), and then add 30 mL of PBS buffer solution with pH=6.5, and keep warm to 40℃.

[0106] (3) Slowly inject 20 mL of ethanol solution into PBS buffer and stir slowly for 90 min.

[0107] (4) Pour the liquid obtained in step (3) into a high-pressure microfluidic device and homogenize it once at a pressure of 30,000 psi to obtain the final product. The sample obtained in this embodiment is a pale yellow transparent liquid.

[0108] Example 3

[0109] Preparation of liposomes with lipase responsive ascorbate tetraisopalmitate

[0110] (1) Weigh out soybean phosphatidylcholine, hydrogenated phosphatidylcholine, cholesterol, PEG-7 olive oil ester, glyceryl stearate, ceramide NP and ascorbate tetraisopalmitate and dissolve them in ethanol solution. The amount of each component is the same as recorded in Table 1. The amount of ascorbate tetraisopalmitate is 0.5g.

[0111] (2) Take a round-bottom flask, add the mixed solution prepared in step (1), and then add 30 mL of PBS buffer solution with pH=7.2, and keep warm to 30℃.

[0112] (3) Slowly inject 20 mL of ethanol solution into PBS buffer and stir slowly for 30 min.

[0113] (4) Pour the above liquid into a high-pressure microjet device and homogenize it at a pressure of 25,000 psi for 5 cycles before discharging. The sample obtained in this embodiment is a pale yellow transparent liquid.

[0114] Example 4

[0115] Preparation of liposomes with lipase responsive glycyrrhizin: The difference from Example 1 is that tau tannin is replaced with an equal amount of glycyrrhizin.

[0116] Example 5

[0117] Preparation of liposomes with lipase responsiveness: The difference from Example 1 is that taureol was replaced with an equal amount of pterostilbene.

[0118] Comparative Example 1

[0119] The difference from Example 1 lies in the change of the lipase-responsive wall material in step (1), as follows:

[0120] (1) Weigh 8g of soybean phosphatidylcholine and 0.3g of cholesterol. The remaining lipids are supplemented with Tween-80 to the same amount of lipids as in Example 1. Dissolve 0.5g of tau phenol in an ethanol solution and heat until it is clear.

[0121] (2) Take a round-bottom flask, add the mixed solution obtained in step (1), and then add 30 mL of PBS buffer solution with pH=6.5, and keep warm to 40℃.

[0122] (3) Slowly inject 20 mL of ethanol solution into PBS buffer and stir slowly for 90 min.

[0123] (4) Pour the liquid obtained in step (3) into a high-pressure microfluidic device and homogenize it once at a pressure of 30,000 psi to obtain the final product. The sample obtained in this embodiment is a pale yellow transparent liquid.

[0124] Comparative Example 2

[0125] The difference from Example 1 is that the high-pressure microjets step in step (4) is replaced with ordinary homogenization, as follows:

[0126] (1) Weigh soybean phosphatidylcholine, hydrogenated phosphatidylcholine, cholesterol, PEG-7 olive oil ester, glyceryl stearate, ceramide NP and tau phenol as recorded in Table 1, dissolve in ethanol solution, and heat until transparent and clear.

[0127] (2) Take a round-bottom flask, add the mixed solution obtained in step (1), and then add 30 mL of PBS buffer solution with pH=6.5, and keep warm to 40℃.

[0128] (3) Slowly inject 20 mL of ethanol solution into PBS buffer and stir slowly for 90 min.

[0129] (4) Homogenize the liquid obtained in step (3) at a speed of 13000 rpm for 10 min, and then discharge the liquid.

[0130] The sample obtained in this comparative example was a pale yellow, opaque liquid. The excessively large particle size caused the sample to separate into layers within 24 hours.

[0131] Comparative Examples 3-5

[0132] The difference from Example 1 is that the amount of each component is shown in Table 3.

[0133] Table 3

[0134]

[0135] Application Example 1

[0136] Preparation of Acne Treatment Containing Peach Phenol Liposomes with Lipase Response

[0137] (1) By weight, add 1% glycerol, 3% butanediol, 0.5% p-hydroxyacetophenone, 0.6% hexanediol, 0.05% sodium hyaluronate and the remainder water to a 200g beaker, and heat at 80℃ for 20min.

[0138] (2) When the temperature of the material drops to 45°C, add 1% of lipase-responsive tau phenol liposomes and the remaining water, and then cool down to obtain the final product.

[0139] Application Example 2

[0140] Preparation of a transparent acne-removing essence containing lipophenol liposomes with lipase responsiveness

[0141] (1) By weight, add 3% glycerol, 5% butanediol, 0.5% p-hydroxyacetophenone, 0.6% hexanediol, 0.05% sodium hyaluronate, 0.1% carbomer 940, 0.1% EDTA-2Na and the remainder water to a 200g beaker, and heat at 80℃ for 20min.

[0142] (2) Homogenize at 7000 rpm for 3 minutes, then stir and cool.

[0143] (3) When the temperature of the material drops to 45°C, add 1% of lipase-responsive tau phenol liposomes and the remainder water. Cool down to obtain the final product.

[0144] Test Example 1

[0145] Liposomes containing lipoprotein exhibiting lipase-responsive cytotoxicity

[0146] The effect of lipoprotein liposomes containing lipase-responsive tau phenol on the cytotoxicity of HaCaT and RAW 264.7 cells was determined by MTT assay: HaCaT or RAW 264.7 cells were used at a cell density of 8000 cells / cm³. 2 Cells were seeded in 96-well plates. Different concentrations of the lipase-responsive tau phenol liposomes prepared in Example 1 were co-cultured with HaCaT cells or RAW 264.7 cells for 48 h. MTT reagent was added, and the cells were incubated for 4 h. DMSO was then added, and the absorbance of each group of cells was measured at 490 nm to calculate the cell viability (%). Figures 1-2 As shown, the results indicated that the cell viability was no less than 85%, indicating that the liposomes containing tau phenol had no cytotoxicity to HaCaT cells and RAW 264.7 cells and had good biosafety.

[0147] The formula for calculating cell viability is as follows:

[0148]

[0149] Where A represents the absorbance value, the negative control group is the absorbance of cells that have not been treated with the drug, the blank control group is the absorbance of the 96-well plate itself, and the sample group is the absorbance of cells treated with the drug in Example 3.

[0150] Test Example 2

[0151] Cumulative permeation assay of transdermal diffusion of lipophenol liposomes and lipophenol suspensions with lipase responsiveness.

[0152] Exfoliated skin from undamaged Bama miniature pigs was cleaned with physiological saline and fixed to a Franz diffusion cell with the cuticle facing upwards. PBS was added to the receiving cell, and after equilibration for 30 minutes, air bubbles were removed by tapping with a rubber bulb. The water bath temperature was set to 32±1℃. 1g of the lipase-responsive tau phenol liposomes prepared in Example 1 was accurately weighed. The receiving cell was stirred at 600 rpm, and samples were collected at 2h, 4h, 6h, 8h, 10h, 12h, and 24h. The samples were centrifuged at 12000 rpm for 10 minutes, and the supernatant was collected. After filtering through a 0.22μm filter membrane, liquid chromatography was performed, and the cumulative permeate was calculated. The diffusion cell diameter was 20mm, and the effective diffusion area was approximately 3.14cm². 2 .

[0153] The formula for calculating cumulative transmittance is as follows:

[0154]

[0155] in, The cumulative permeation rate is given by V, where V is the total volume of the receiving liquid. The concentration of the drug in the receiving solution at the nth sampling time. Let be the mass concentration of the drug in the receiving solution during the i-th sampling. Where A is the sampling volume and A is the effective diffusion area.

[0156] The experimental results are shown in Figure 3 .

[0157] Test Example 3

[0158] Lipase-responsive test of taurine liposomes

[0159] A dialysis bag with a permeability of 1000 Da was selected and completely immersed in water. 8 mL of PBS, 1 mL of lipase-responsive tau phenol liposomes prepared in Examples 1, 3-5, and 1 mL of lipase solution with an enzyme activity of 200 U / g were added to the dialysis bag. No lipase was added to the blank control group. The dialysis bag was completely sealed and placed in a beaker, with 30% ethanol solution added as the receiving solution. The water bath temperature was set to 37±1℃, and the stirring speed of the receiving cell was set to 600 rpm. Samples were collected from the receiving cell at 1h, 2h, 4h, 8h, 12h, 24h, and 48h, centrifuged at 12000 rpm for 10 min, and the supernatant was collected, filtered through a 0.22 μm filter, and then subjected to UV spectrophotometry to calculate the cumulative release. Figures 4-9 As shown, ordinary tau phenol liposomes are not sensitive to lipase response, while the tau phenol liposomes prepared in Example 1 have obvious lipase response.

[0160] like Figure 9As can be seen, Example 1 exhibits good lipase responsiveness, reaching a release plateau within 48 hours with an overall release rate greater than 60%, and showing no significant burst release in the first 4 hours. Comparative Example 3, by removing ceramide, resulted in a burst release within 2 hours; Comparative Example 4, by removing glyceryl stearate, caused a significant decrease in lipase responsiveness during liposome release, resulting in a large amount of drug not being released; Comparative Example 5, by removing PEG-7 olive oil ester, also resulted in a burst release within 2 hours, but its burst release rate was lower than that of Comparative Example 3.

[0161] Test Example 4

[0162] Antibacterial efficacy test of lipophenol liposomes with lipase responsiveness

[0163] Select bacterial culture in the logarithmic growth phase and dilute it to 10⁻⁶ with physiological saline. 8 CFU / mL, bacterial suspension and sample solution were added separately to 96-well plates, with each sample concentration repeated three times. The blank control contained no sample solution. The mixed 96-well plates or culture flasks were incubated at 37 ºC on a shaker at 250 rpm for 24 h. 1 mL of the test sample was collected at 3 h, 6 h, 12 h, and 24 h for colony count testing.

[0164] The experimental results are shown in Figures 10-11 .

[0165] Test Example 5

[0166] Test on the effect of lipase-responsive tau phenol liposomes on linoleic acid peroxidation

[0167] Take an appropriate amount of lipase-responsive tau phenol liposomes, dilute them with ethanol-water solutions according to a concentration gradient, add them to the sample containing linoleic acid, stir well, and adjust the pH to 7. Store each group of samples in a 45℃ oven, and take samples at 1, 2, 3, 5, and 10 days. Measure the absorbance using the ammonium thiocyanate and ferric ion colorimetric method, and record the absorbance at each time point to characterize the degree of linoleic acid oxidation.

[0168] The experimental results are shown in Figure 12 .

[0169] Test Example 6

[0170] Stability analysis of lipoprotein liposomes with lipase responsiveness.

[0171] After diluting an appropriate amount of lipase-responsive tau phenol liposomes, the samples were tested using a LUM analyzer with the following settings: rotation speed 2500 rpm, time interval 45 s, temperature 45 ℃, and wavelength 870 nm.

[0172] The experimental results are shown in Figure 13 .

[0173] Test Example 7

[0174] Stability test

[0175] Test sample: The sample prepared in Example 1.

[0176] Experimental methods:

[0177] Each test sample was tested for 7 days, 14 days, 21 days, and 28 days under five test conditions: -18℃, room temperature, light irradiation, 45℃, and 4℃. The changes in particle size and encapsulation efficiency of each test sample were measured.

[0178] The experimental results are shown in Figure 14-15 .

[0179] The above detailed description is a specific description of one of the feasible embodiments of the present invention. This embodiment is not intended to limit the patent scope of the present invention. All equivalent implementations or modifications that do not depart from the present invention should be included within the scope of the technical solution of the present invention.

Claims

1. A liposome with lipase responsiveness, comprising a wall material and a core material, characterized in that, The wall material is composed of the following components: 8-16 parts soybean phosphatidylcholine, 0.5-1.5 parts hydrogenated phosphatidylcholine, 0.3-0.8 parts cholesterol, 2-3 parts PEG-7 olive oil ester, 2-3 parts glyceryl stearate, and 2-3 parts ceramide NP; The core material is selected from at least one of the following: tau phenol, paeonol, magnolol, capryloyl glycine, glycyrrhizin, ascorbate tetraisopalmitate, tetrahydrocurcumin, or pterostilbene. The ratio of the core material to the wall material by weight is 1:30 to 1:

50.

2. The use of the liposomes according to claim 1 in the preparation of cosmetics or pharmaceuticals, wherein the cosmetics include acne-removing lotions, acne-removing essences, lotions, and creams; and the pharmaceuticals include gels or ointments.

3. A product characterized in that, The product includes the liposomes described in claim 1; the product is a cosmetic or a pharmaceutical.

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