Biomimetic vesicle with controllable rigidity and flexibility of membrane, preparation method and application

Abstract

The application provides a biomimetic vesicle with controllable membrane rigidity and flexibility, a preparation method and application, and comprises the following steps: mixing a membrane rigidity adjusting component and a membrane flexibility adjusting component according to a design ratio, stirring under heating conditions until complete melting and uniformity; adding an aqueous phase in which a hydrophilic active ingredient is dissolved into the mixed system, heating and stirring to form a primary emulsion; supplementing deionized water into the primary emulsion to a target mass, and carrying out high shear homogenization and pulse ultrasonic treatment to obtain the biomimetic vesicle; the membrane rigidity adjusting component comprises ceramide and cholesterol; the membrane flexibility adjusting component comprises soybean phospholipid, DLGL and MEL-B; the ratio among MEL-B, DLGL, soybean phospholipid, ceramide and cholesterol is 5:1:0.75:0.375:0.1875; the biomimetic vesicle obtained by the application has deep and persistent retention and absorption effects in skin penetration in vivo, is superior to free active ingredients, and has the development value of cosmetic and transdermal drug delivery preparations.

Description

Technical Field

[0001] This invention belongs to the technical fields of transdermal drug delivery and cosmetics, specifically relating to a biomimetic vesicle with controllable membrane rigidity and flexibility, its preparation method, and its application. Background Technology

[0002] Hydrophilic active ingredients (such as polysaccharides, oligosaccharides, peptides, and vitamin C derivatives) have difficulty penetrating the skin due to the strong barrier effect of the stratum corneum, resulting in low bioavailability. Vesicle-like carriers (such as liposomes and delivery systems) have become effective tools for promoting transdermal penetration due to their bilayer structure similar to biological membranes. However, current technologies mostly focus on optimizing vesicle components or comparing vesicle types, lacking systematic methods for regulating the physical properties of vesicle membranes (especially rigidity and flexibility), and failing to establish a quantitative structure-activity relationship between membrane rigidity / flexibility and its transdermal performance.

[0003] Chinese patent CN201610734381.5 discloses a self-assembled carrier of mannose-erythritol lipids and lecithin, but it only focuses on thermal stability and does not address the regulation of membrane rigidity and flexibility. Another type of biomimetic vesicle (such as CN117598915A) introduces stratum corneum lipid components such as ceramides and cholesterol, but it only remains at the level of component imitation and does not reveal the intrinsic relationship between membrane rigidity and flexibility and permeation efficiency. Therefore, there is an urgent need for a method that can quantitatively regulate membrane rigidity and flexibility and accurately predict transdermal performance. Summary of the Invention

[0004] To address the aforementioned technical problems, this invention provides biomimetic vesicles with controllable membrane rigidity and flexibility, their preparation method, and their applications.

[0005] On the one hand, this invention provides the following technical solution: a method for preparing biomimetic vesicles with controllable membrane rigidity and flexibility, the method comprising: The membrane rigidity regulating component and the membrane flexibility regulating component are mixed in the designed ratio and stirred under heating conditions until completely melted and homogeneous to obtain a mixed system; Add an aqueous phase containing dissolved hydrophilic active ingredients to the mixture, and continue heating and stirring to form a primary emulsion; Deionized water was added to the primary emulsion to the target mass, and the emulsion was subjected to high-shear homogenization and pulsed ultrasonic treatment in sequence to obtain biomimetic vesicles with a polydispersity index (PDI) of less than 0.3. The membrane rigidity regulating component includes ceramide and cholesterol, and the membrane flexibility regulating component includes soybean lecithin, DLGL, and MEL-B. The ratio of MEL-B, DLGL, soybean lecithin, ceramide, and cholesterol is (4-6):1:(0.5-1):(0.125-0.75):(0.0625-0.5).

[0006] The beneficial effects of the method for preparing biomimetic vesicles with controllable membrane rigidity and flexibility proposed in this invention are as follows: The first proposal to use the intensity ratio of Raman spectral characteristic peaks I 2880 / I 2850 As a quantitative control index of the rigidity and flexibility of biomimetic vesicle membranes, a predictable structure-activity relationship of "component ratio-membrane rigidity and flexibility-transdermal efficiency" was established, providing a scientific basis for the rational design of transdermal carriers. When the membrane stiffness and flexibility are at their optimal value (I) 2880 / I 2850 When the concentration is 1.14, the cumulative permeate over 8 hours can reach 127.7 μg / cm³. 2 Its relative penetration rate is about 4 times that of free active ingredients; It should be noted that this invention further discovers that the membrane's stiffness and flexibility are not necessarily better the more flexible or the more rigid it is, but rather there exists an optimal range (I... 2880 / I 2850 The Raman ratio is 1.12–1.16. When the ratio is below 1.12 (membrane too flexible), the stability of the vesicle structure decreases, making it prone to rupture on the skin surface or within the stratum corneum, leading to premature leakage of the encapsulated hydrophilic active ingredients. When the ratio is above 1.16 (membrane too rigid), the vesicles lack deformability and have difficulty penetrating the dense lipid channels of the stratum corneum. Only when the Raman ratio is precisely controlled between 1.12 and 1.16 can the vesicles possess both sufficient structural integrity and suitable deformability, with a more preferred range of 1.13–1.14, within which deep and long-lasting transdermal retention and absorption can be achieved. The preparation method is simple, reproducible, applicable to a variety of hydrophilic active ingredients, and easy to scale up for production. The resulting biomimetic vesicles exhibit deep and long-lasting retention and absorption effects during in vivo skin penetration, which is superior to free active ingredients, and have development value in both cosmetics and transdermal drug delivery formulations.

[0007] In addition, the method for preparing biomimetic vesicles with controllable membrane rigidity and flexibility according to the present invention may also have the following additional technical features: The preferred ratio of MEL-B, DLGL, soy lecithin, ceramide, and cholesterol is 5:1:0.75:0.375:0.1875.

[0008] Preferably, the hydrophilic active ingredient includes one or more of Dendrobium oligosaccharides, polysaccharides, oligopeptides, and vitamin C derivatives.

[0009] On the other hand, the invention provides the following technical solution: a biomimetic vesicle with controllable membrane rigidity and flexibility, wherein the biomimetic vesicle is prepared by the above-described method for preparing a biomimetic vesicle with controllable membrane rigidity and flexibility.

[0010] Preferably, the Raman spectral characteristic peak intensity of the biomimetic vesicle is higher than that of I. 2880 / I 2850 Between 1.12 and 1.16.

[0011] Preferably, the Raman spectral characteristic peak intensity of the biomimetic vesicle is higher than that of I. 2880 / I 2850 Specifically, it is between 1.13 and 1.14.

[0012] On the other hand, the invention provides the following technical solution: the application of a biomimetic vesicle with controllable membrane rigidity and flexibility as described above in the preparation of cosmetics or drugs that promote the transdermal penetration of hydrophilic active ingredients.

[0013] Preferably, the hydrophilic active ingredient includes one or more of Dendrobium oligosaccharides, polysaccharides, oligopeptides, and vitamin C derivatives. Attached Figure Description

[0014] To more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0015] Figure 1 This is a transmission electron microscope image of the vesicles prepared in Example 1 of the present invention; Figure 2 The Raman spectrum of the vesicles prepared in Example 1 of this invention; Figure 3 The in vivo permeability analysis results of the vesicle solutions prepared in Example 1, which are Dendrobium oligosaccharides and Dendrobium oligosaccharide-loaded solutions.

[0016] The present invention will be further described below with reference to the accompanying drawings and description. Detailed Implementation

[0017] Embodiments of the present invention are described in detail below, examples of which are illustrated in the accompanying drawings, wherein the same or similar reference numerals denote the same or similar elements or elements having the same or similar functions throughout. The embodiments described below with reference to the accompanying drawings are exemplary and intended to explain embodiments of the present invention, and should not be construed as limiting the present invention.

[0018] This invention provides a biomimetic vesicle with controllable membrane rigidity and flexibility, its preparation method, and its application. Those skilled in the art can refer to the content of this document and appropriately modify the process parameters to achieve the desired result. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art and are considered to be included in this invention. The method and application of this invention have been described through preferred embodiments, and those skilled in the art can clearly modify or appropriately change and combine the method and application described herein without departing from the content, spirit, and scope of this invention to realize and apply the technology of this invention.

[0019] The method for preparing the rigid-flexible controllable biomimetic vesicles of the present invention includes: The membrane rigidity regulating component and the membrane flexibility regulating component are mixed in the designed ratio and stirred under heating conditions until completely melted and homogeneous to obtain a mixed system; Add an aqueous phase containing dissolved hydrophilic active ingredients to the mixture, and continue heating and stirring to form a primary emulsion; Deionized water was added to the primary emulsion to the target mass, and the emulsion was subjected to high-shear homogenization and pulsed ultrasonic treatment in sequence to obtain biomimetic vesicles with a polydispersity index (PDI) of less than 0.3. The membrane rigidity regulating component includes ceramide and cholesterol, and the membrane flexibility regulating component includes soybean lecithin, DLGL, and MEL-B. The ratio of MEL-B, DLGL, soybean lecithin, ceramide, and cholesterol is (4-6):1:(0.5-1):(0.125-0.75):(0.0625-0.5).

[0020] Among them, DLGL is sodium bis(lauramide-glutamine)lysine, and MEL-B is mannoerythritol ester.

[0021] Specifically, the ratio of MEL-B, DLGL, soy lecithin, ceramide, and cholesterol is 5:1:0.75:0.375:0.1875.

[0022] Specifically, the hydrophilic active ingredients include one or more of Dendrobium oligosaccharides, polysaccharides, oligopeptides, and vitamin C derivatives.

[0023] For the biomimetic vesicles of the present invention, the biomimetic vesicles are prepared by the biomimetic vesicle preparation method with controllable membrane stiffness and flexibility as described above.

[0024] Among them, the Raman spectral characteristic peak intensity of the biomimetic vesicle is higher than that of I. 2880 / I 2850 The value is between 1.12 and 1.16, preferably between 1.13 and 1.14, to achieve the best transdermal penetration efficiency.

[0025] In terms of the application of the present invention, a biomimetic vesicle with controllable membrane rigidity and flexibility as described above is used in the preparation of cosmetics or pharmaceuticals that promote the transdermal penetration of hydrophilic active ingredients.

[0026] The hydrophilic active ingredients include one or more of the following: Dendrobium oligosaccharides, polysaccharides, oligopeptides, and vitamin C derivatives.

[0027] Specifically, to further verify the effectiveness of the present invention, the following experiments will demonstrate it: 1. Component Experiment Example 1 This embodiment provides a biomimetic vesicle with controllable membrane rigidity and flexibility. The preparation method specifically includes: grinding flaky cholesterol (57-88-5, AR, Shanghai Yuanye Biotechnology Co., Ltd.) into powder using a mortar and pestle. Weigh 0.30g of ground cholesterol, 0.60g of ceramide (100403-19-8, ≥ 95%, Adamas-beta), and 8.00g of mannose erythropoiesis lipolipase (MEL-B, Shanghai Puen Biochemical Technology Co., Ltd.) into a 150 mL beaker. Add a magnetic stir bar and stir in a water bath at 70°C for 10 min to ensure the cholesterol, ceramide, and MEL-B are evenly mixed. Then add 1.20g of soybean lecithin (PC50, Shanghai Aikon Fine Chemical Co., Ltd.) and an aqueous solution of DLGL containing 1.60g of pure DLGL (e.g., weigh 5.52g of 29wt% DLGL aqueous solution, Shanghai Kunyang Industrial Co., Ltd.). Continue stirring at 70°C for 5 min. After complete mixing, add deionized water containing 25.0g of Dendrobium oligosaccharide (hydrophilic active ingredient) and continue stirring in a water bath for 5 min. After the solution showed no solid precipitation, sufficient deionized water was added to bring the total mass to 1000g. Stirring continued for 10 minutes until the entire system was uniformly dispersed. The mixture was then removed and cooled to room temperature. A high-shear homogenizer was used to homogenize the solution at 10000 rpm for 2 minutes, followed by ultrasonic dispersion emulsifier operation in pulse mode for 300 seconds (duty cycle 40%, ON / OFF = 2 s / 3 s, power 300 W). This yielded biomimetic vesicle samples with a polydispersity index (PDI) less than 0.3 (the average particle size of the vesicles was determined using dynamic light scattering, measured using a NanoBrook Omni Zeta potential and nanoparticle size analyzer manufactured by Brookhaven Instruments, USA). (Note: In subsequent examples and comparative examples, the preparation process and parameters remained unchanged, but the amount of added components varied.)

[0028] Example 2 This embodiment provides a biomimetic vesicle with controllable membrane rigidity and flexibility. Its preparation method is roughly the same as that of Embodiment 1, except that the amount of MEL-B added is 6.4g.

[0029] Example 3 This embodiment provides a biomimetic vesicle with controllable membrane rigidity and flexibility. Its preparation method is roughly the same as that of Embodiment 1, except that the amount of MEL-B added is 9.6g.

[0030] Example 4 This embodiment provides a biomimetic vesicle with controllable membrane rigidity and flexibility. Its preparation method is roughly the same as that of Embodiment 1, except that the amount of soybean lecithin added is 0.8g.

[0031] Example 5 This embodiment provides a biomimetic vesicle with controllable membrane rigidity and flexibility. Its preparation method is roughly the same as that of Embodiment 1, except that the amount of soybean lecithin added is 1.6g.

[0032] Example 6 This embodiment provides a biomimetic vesicle with controllable membrane rigidity and flexibility. Its preparation method is roughly the same as that of Embodiment 1, except that the amount of ceramide added is 0.2g.

[0033] Example 7 This embodiment provides a biomimetic vesicle with controllable membrane rigidity and flexibility. Its preparation method is roughly the same as that of Embodiment 1, except that the amount of ceramide added is 1.2g.

[0034] Example 8 This embodiment provides a biomimetic vesicle with controllable membrane rigidity and flexibility. Its preparation method is roughly the same as that of Embodiment 1, except that the amount of cholesterol added is 0.1g.

[0035] Example 9 This embodiment provides a biomimetic vesicle with controllable membrane rigidity and flexibility. Its preparation method is roughly the same as that of Embodiment 1, except that the amount of cholesterol added is 0.8g.

[0036] Example 10 This embodiment provides a biomimetic vesicle with controllable membrane rigidity and flexibility. Its preparation method is roughly the same as that of Embodiment 1, except that the amount of soybean lecithin added is 0.8g, the amount of ceramide added is 0.2g, and the amount of cholesterol added is 0.1g.

[0037] Example 11 This embodiment provides a biomimetic vesicle with controllable membrane rigidity and flexibility. Its preparation method is roughly the same as that of Embodiment 1, except that the amount of soybean lecithin added is 1.6g, the amount of ceramide added is 1.2g, and the amount of cholesterol added is 0.8g.

[0038] Comparative Example 1 This comparative example provides a biomimetic vesicle with controllable membrane rigidity and flexibility. Its preparation method is roughly the same as that of Example 1, except that MEL-B is not added.

[0039] Comparative Example 2 This comparative example provides a biomimetic vesicle with controllable membrane rigidity and flexibility. Its preparation method is roughly the same as that of Example 1, except that DLGL is not added.

[0040] Comparative Example 3 This comparative example provides a biomimetic vesicle with controllable membrane rigidity and flexibility. Its preparation method is roughly the same as that of Example 1, except that soybean lecithin is not added.

[0041] Comparative Example 4 This comparative example provides a biomimetic vesicle with controllable membrane rigidity and flexibility. Its preparation method is roughly the same as that of Example 1, except that no ceramide is added.

[0042] Comparative Example 5 This comparative example provides a biomimetic vesicle with controllable membrane rigidity and flexibility. Its preparation method is roughly the same as that of Example 1, except that cholesterol is not added.

[0043] Comparative Example 6 This comparative example provides a biomimetic vesicle with controllable membrane rigidity and flexibility. Its preparation method is roughly the same as that of Example 1, except that MEL-B and DLGL are not added.

[0044] Comparative Example 7 This comparative example provides a biomimetic vesicle with controllable membrane rigidity and flexibility. Its preparation method is roughly the same as that of Example 1, except that soybean lecithin, ceramide, and cholesterol are not added.

[0045] 2. In vitro percutaneous permeation test In vitro permeation experiments were performed using the Franz diffusion method. Piglet skin that had been treated with grease was pretreated and soaked in PBS buffer for 30 min. After treatment, the piglet skin was cut into pieces approximately 4 cm in size using a scalpel. 2 The apparatus was circular. The Franz receiving cell was filled with 7 mL of PBS buffer and placed into the matching rotor. The cut piglet skin was then carefully placed between the diffusion and receiving cells, with the epidermis facing the diffusion cell and the dermis facing the receiving cell. During assembly, all air bubbles in the receiving solution must be expelled to avoid affecting the transdermal effect. The apparatus was then secured with clamps. 1 mL of the sample to be tested was added to the drug delivery chamber, and the chamber was sealed with plastic wrap to prevent solvent evaporation during the experiment. During the experiment, the transdermal diffusion apparatus (RYJ-12B, Shanghai Huanghai Pharmaceutical Testing Instruments Co., Ltd.) was maintained at (37 ± 0.5) °C, the stirring rate was set to 300 rpm, and the transdermal duration was 8 h.

[0046] The biomimetic vesicle samples prepared in the above examples and comparative examples were subjected to in vitro percutaneous permeation experiments and stability experiments (the prepared samples were placed in glass containers and left at room temperature for 1 month; the changes in the samples were observed. If precipitation or white turbidity appeared in the samples, they were determined to be unstable). The results are shown in Table 1 below: Table 1

[0047] As shown in Table 1 above, this invention, for the first time, uses the intensity ratio of Raman spectral characteristic peaks (I... 2880 / I 2850 MEL was systematically investigated as a quantitative control index of the rigidity and flexibility of biomimetic vesicle membranes. The effects of the proportions of five components—B, DLGL, soybean lecithin, ceramide, and cholesterol—on the physical properties and transdermal permeability of vesicle membranes were successfully established, establishing a predictable structure-activity relationship of "component ratio—membrane stiffness / flexibility—transdermal efficiency." Table 1 shows that the transdermal permeation-enhancing effects of all example groups were significantly better than those of the comparative groups, with Example 1 exhibiting the best performance, achieving a cumulative permeation of 127.7 ± 5.0 μg / cm³ over 8 hours. 2 The relative permeability of the free active ingredient (the cumulative permeation of free Dendrobium oligosaccharides over 8 hours was 31.2 ± 3.1 μg / cm³) was [not specified]. 2 4.09 times that of ), and the Raman ratio I 2880 / I 2850 The value is 1.14, which falls exactly within the optimal range (1.13 to 1.14) discovered in this invention.

[0048] Comparative Examples 1 to 7 lacked one or more of the following components: MEL-B, DLGL, soybean lecithin, ceramide, or cholesterol. Their 8-hour cumulative permeation was significantly lower than that of the Example Group (maximum only 70.2 μg / cm³). 2 Furthermore, most comparative samples exhibited poor stability or Raman ratios deviating from the ideal range (1.12–1.16). The specific reasons are analyzed below: (1) The synergistic effect of the five components is the basis for the integrity of vesicle function. MEL B and DLGL, as amphiphilic surfactants, together construct the bilayer framework of vesicles. The combination of these two surfactants has a certain permeation-enhancing effect, but they cannot form a stable and efficient drug delivery system when present alone. MEL is lacking. When B (Comparative Example 1), DLGL (Comparative Example 2), or both are absent (Comparative Example 6), the system cannot form complete vesicles, resulting in a significant decrease in transdermal efficiency (all below 35.1 μg / cm³). 2 Furthermore, the sample is unstable.

[0049] (2) Soybean lecithin, ceramide, and cholesterol jointly regulate the rigidity and stability of the membrane. Soybean lecithin imparts suitable fluidity and deformability to the membrane, and is a key component for flexibility regulation; ceramide and cholesterol enhance the orderliness and rigidity of the membrane through hydrogen bonding and hydrophobic interactions, improving the structural integrity of vesicles in the stratum corneum of the skin. Comparative Example 3 (without soybean lecithin) had a Raman ratio of 1.17, indicating not only that the vesicle membrane was too rigid, resulting in poor transdermal effect, but also that the lack of soybean lecithin prevented ceramide and cholesterol from forming a stable bilayer membrane with MEL-B and DLGL, ultimately leading to sample instability; the Raman ratios of Comparative Example 4 (without ceramide) and Comparative Example 5 (without cholesterol) were 1.15 and 1.13, respectively, both falling within the range of 1.12 to 1.16 described in this invention, and even approaching the preferred range of 1.13 to 1.14. However, its 8-hour cumulative permeation (60.1 μg / cm) was low. 2 and 70.2 μg / cm 2 Compared to Example 1 (127.7 μg / cm) 2 The percentages of ceramides and cholesterol decreased by 53% and 45% respectively, showing a significant reduction. This result indicates that simply meeting the Raman ratio (membrane rigidity / flexibility) is insufficient to achieve optimal transdermal efficiency. Besides regulating membrane rigidity / flexibility, ceramides and cholesterol also share homology with natural lipids in the stratum corneum, promoting the fusion of vesicles with stratum corneum lipids and enhancing the deep delivery of active ingredients. Without any one of these components, even if the vesicle rigidity / flexibility parameters are satisfactory, its compatibility with the skin and permeation efficiency cannot reach the optimal level of synergistic effects among the five components. Therefore, ceramides and cholesterol are essential components for achieving high-efficiency transdermal absorption in this biomimetic vesicle; all five components are indispensable.

[0050] (3) The precise control of membrane stiffness and flexibility, as well as the ratio of synergistic components, jointly affects transdermal efficiency. Comparative Example 7 (lacking soybean lecithin, ceramide, and cholesterol) had a Raman ratio of 1.10 (membrane too flexible), indicating that vesicles were prone to rupture and had weak interaction with stratum corneum lipids; Comparative Example 3 (without soybean lecithin) had a Raman ratio of 1.17 (membrane too rigid), indicating that vesicles were difficult to deform and could not effectively penetrate the skin. Neither of these methods could achieve efficient transdermal delivery, which further confirms that transdermal efficiency can only be maximized when the membrane stiffness and flexibility are within the optimal range (1.12–1.16, preferably 1.13–1.14) discovered in this invention and the ratio of synergistic components falls within the optimal range.

[0051] (4) Soybean phospholipids, ceramides, and cholesterol are all natural lipid components in the stratum corneum. By optimizing the ratio of the five components, this invention can simulate the lipid environment of the stratum corneum, thereby maximizing the synergistic permeation-enhancing effect. This synergistic effect further verifies the necessity of the "dual control of ratio and rigidity" mentioned in point (3). The absence of any single component (Comparative Examples 1-5) or the combined absence of key components (Comparative Examples 6 and 7) cannot achieve the balance between transdermal efficiency and stability as described in Example 1.

[0052] In all embodiments, Embodiment 1 (MEL) B : DLGL : soybean phospholipids : ceramides : cholesterol = 8 : 1.6 : 1.2 : 0.6 : 0.3 (mass ratio 80 : 16 : 12 : 6 : 3) showed the most outstanding performance. Its 8-hour cumulative permeate reached 127.7 ± 5.0 μg / cm³. 2 The relative permeability was 3.02 times that of Comparative Example 7; the sample was stable after being stored at room temperature for one month, and the Raman ratio I... 2880 / I 2850 = 1.14, which falls precisely within the optimal range (1.13–1.14). At this ratio, the vesicles achieve a precise balance between rigidity and flexibility, and optimal synergy among the five components: the orderliness and fluidity of the membrane lipid chains are optimally matched, ensuring that the vesicles possess sufficient deformability within the stratum corneum of the skin (see...). Figure 1 It has an elliptical, deformable shape, which maintains the integrity of the membrane structure and prevents premature leakage of contents. Specifically: MEL The mass ratio of B to DLGL (5:1) ensures the stability of the basic vesicle structure; the mass ratio of soybean lecithin to DLGL (0.75) provides suitable flexibility; and the mass ratio of ceramide to cholesterol (2:1) and their total mass to the mass ratio of soybean lecithin (0.75) together construct an ideal rigid support. This combination enables the vesicles to exhibit a moderate ratio of CH2 symmetric to asymmetric stretching vibration peak intensity in Raman spectroscopy, reflecting the optimal order of the membrane lipid chain arrangement.

[0053] In summary, existing technologies only focus on vesicle component mimicry or stability, failing to reveal the quantitative relationship between membrane rigidity / flexibility and transdermal efficiency. This invention is the first to propose using the Raman characteristic peak intensity ratio I... 2880 / I 2850 As a quantitative indicator of membrane stiffness and flexibility, and defining the specific ratio range of the five components (MEL) B. The ratio of DLGL, soybean lecithin, ceramide, and cholesterol is (4) 6):1:(0.5 1):(0.125 0.75):(0.0625 0.5), among which the optimal ratio (5:1:0.75:0.375:0.1875) can stably obtain I 2880 / I 2850 Vesicles with a density of 1.14 have a transdermal transdermal efficiency approximately four times that of free active ingredients. Specifically, this is achieved when the membrane stiffness and flexibility are within the optimal range (I...). 2880 / I 2850 When the ratio of the five components is 1.13 to 1.14 and falls within the preferred range defined by this invention, the optimal transdermal efficiency can be achieved. Comparative examples and other embodiments cannot simultaneously satisfy the requirements of "stable vesicle structure" and "moderate membrane stiffness and flexibility (I...)". 2880 / I 2850 The three core requirements of "1.13-1.14" and "the proportion of synergistic components falls within the optimal range" make it impossible to achieve optimal transdermal efficiency.

[0054] 3. Cryo-transmission electron microscopy observation The vesicle solution prepared in Example 1 was dropped onto a hydrophilically treated copper mesh. Excess liquid at the edges was absorbed with filter paper. The copper mesh was then immersed in liquid ethane pre-cooled to -180°C, rapidly frozen, and transferred to a grid box for observation under an electron microscope. Figure 1 As shown. The morphology of the prepared vesicle membrane can be directly observed using a Cryo-TEM (FEI Talos F200C, Thermo Fisher Scientific). Vesicles with high membrane fluidity are more deformable and elliptical, while vesicles with high membrane rigidity tend to form standard spherical shapes.

[0055] 4. Raman spectroscopy determination experiment Beforehand, cut aluminum foil to a suitable size and wrap it around the special glass slide of the Raman spectrometer (LabRAM Soleil, HORIBA, China). Use a pipette to take 2 μL of the vesicle solution to be tested onto the aluminum foil. The measurement was performed using a laser excitation wavelength of 532 nm, a laser power of 17 mW, and an exposure time of 3 s. The number of integrations was 3.

[0056] The above experiment shows that, as Figure 2 As shown, the I of the sample prepared in Example 1 2880 / I 2850 =1.14.

[0057] 5. Characterizing the transdermal permeability of Dendrobium oligosaccharides in vivo Before testing the in vivo permeation of Dendrobium oligosaccharides using a high-resolution, ultra-sensitive intelligent Raman imager (LabRAM Soleil, HORIBA, China), the raw materials were scanned. Measurements were performed using a 532 nm laser excitation wavelength, 42 mW laser power, and an exposure time of 2 s, with 3 scans. The in vivo permeation behavior of the vesicle solution and free Dendrobium oligosaccharide solution prepared in Example 1 loaded with Dendrobium oligosaccharides was tested using the Raman imager. A scan was performed before applying the vesicle solution and free Dendrobium oligosaccharide solution prepared in Example 1 to the inner arm as a blank control. Two areas of equal size were marked on the inner arm, and 300 μL of the vesicle solution prepared in Example 1 and the free Dendrobium oligosaccharide solution were applied to each area, respectively. To ensure the same permeation time for the samples during scanning, the application time was spaced 15 min apart. The permeated areas were scanned using the Raman imager at 0, 2, 4, and 8 h of permeation. Measurements were performed using a 532 nm laser excitation wavelength, a 1.3 mW laser power, and an exposure time of 1 s, with 3 scans.

[0058] The above experiment shows that, as Figure 3 As shown, in the free Dendrobium oligosaccharide group, a weak characteristic signal was detected only in the stratum corneum region 4 h after administration. Eight h after administration, the signal was detected at the junction of the stratum corneum and the active epidermal layer, but the intensity remained low, indicating that the free Dendrobium oligosaccharides mainly remained on the skin surface with limited penetration into deeper layers. In contrast, the vesicle solution group prepared in Example 1, loaded with Dendrobium oligosaccharides, exhibited a significant transdermal penetration advantage. Four h after administration, a strong Dendrobium oligosaccharide signal was detected not only in the stratum corneum but also clearly distributed in the active epidermal layer, with some signals reaching deep into the active epidermis. Eight h after administration, the Dendrobium oligosaccharide signal further enhanced, showing a strong distribution throughout the entire epidermal layer, and the penetration depth significantly exceeded that of the free Dendrobium oligosaccharide group. These results indicate that using biomimetic vesicles as a carrier can significantly improve the transdermal penetration efficiency of Dendrobium oligosaccharides in human skin. The in vivo permeation results are consistent with the previous in vitro porcine skin permeation experiments, jointly confirming that the vesicle solution prepared in Example 1 loaded with Dendrobium oligosaccharides can effectively overcome the skin barrier and efficiently deliver the hydrophilic active ingredient Dendrobium oligosaccharides into the skin, reaching the deep active epidermal layer.

[0059] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention are equivalent substitutions and are included within the protection scope of the present invention.

Claims

1. A method for preparing biomimetic vesicles with controllable membrane rigidity and flexibility, characterized in that, The method includes: The membrane rigidity regulating component and the membrane flexibility regulating component are mixed in the designed ratio and stirred under heating conditions until completely melted and homogeneous to obtain a mixed system; Add an aqueous phase containing dissolved hydrophilic active ingredients to the mixture, and continue heating and stirring to form a primary emulsion; Deionized water was added to the primary emulsion to the target mass, and the emulsion was subjected to high-shear homogenization and pulsed ultrasonic treatment in sequence to obtain biomimetic vesicles with a polydispersity index (PDI) of less than 0.

3. The membrane rigidity regulating component includes ceramide and cholesterol, and the membrane flexibility regulating component includes soybean lecithin, DLGL, and MEL-B. The ratio of MEL-B, DLGL, soybean lecithin, ceramide, and cholesterol is (4-6):1:(0.5-1):(0.125-0.75):(0.0625-0.5).

2. The method for preparing biomimetic vesicles with controllable membrane rigidity and flexibility according to claim 1, characterized in that, The mass ratio of MEL-B, DLGL, soybean lecithin, ceramide, and cholesterol is 5:1:0.75:0.375:0.1875.

3. The method for preparing biomimetic vesicles with controllable membrane rigidity and flexibility according to claim 1, characterized in that, The hydrophilic active ingredients include one or more of the following: Dendrobium oligosaccharides, polysaccharides, oligopeptides, and vitamin C derivatives.

4. A biomimetic vesicle with controllable membrane rigidity and flexibility, wherein the biomimetic vesicle is prepared by the preparation method of the biomimetic vesicle with controllable membrane rigidity and flexibility as described in any one of claims 1-3.

5. The biomimetic vesicle with controllable membrane rigidity and flexibility according to claim 4, characterized in that, The Raman spectral characteristic peak intensity ratio of the biomimetic vesicle is I 2880 / I 2850 Between 1.12 and 1.

16.

6. The biomimetic vesicle with controllable membrane rigidity and flexibility according to claim 5, characterized in that, The Raman spectral characteristic peak intensity ratio of the biomimetic vesicle is I 2880 / I 2850 Specifically, it is between 1.13 and 1.

14.

7. The application of a biomimetic vesicle with controllable membrane rigidity and flexibility as described in any one of claims 4-6 in the preparation of cosmetics or pharmaceuticals that promote the transdermal penetration of hydrophilic active ingredients.

8. The application according to claim 7, characterized in that, The hydrophilic active ingredients include one or more of the following: Dendrobium oligosaccharides, polysaccharides, oligopeptides, and vitamin C derivatives.

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