Transmembrane drug delivery system and preparation method and application thereof

Abstract

The invention discloses a transmembrane drug-loaded delivery system and a preparation method and application thereof, and particularly relates to the technical field of aquaculture disease prevention and control. Wherein the transmembrane drug-loading delivery system is formed by compounding a polyphenol compound with anti-parasitic activity, a drug carrier and a penetration enhancer. Meanwhile, the invention also discloses a preparation method of the transmembrane drug-loaded delivery system, and the method comprises the following steps: adding the polyphenol compound into the drug carrier, and stirring in a water bath until the polyphenol compound is completely dissolved to prepare a drug-containing oil phase; mixing the permeation promoter A, the permeation promoter B and the penetration enhancer, and uniformly stirring to obtain a mixed phase; fully mixing the medicine-containing oil phase with the mixed phase to form a premix; and adding the premix into ultrapure water which is preheated to 35-40 DEG C, and continuously stirring to obtain the transmembrane drug-loading delivery system. The transmembrane drug-loading delivery system can permeate host mucus and a parasite barrier, so that effective components are accurately delivered to a target spot, and the effect of killing parasites in fish bodies is achieved.

Description

Technical Field

[0001] This application relates to the field of aquaculture disease prevention and control technology, specifically to a transmembrane drug delivery system and its preparation method and application. Background Technology

[0002] In intensive freshwater aquaculture, parasitic diseases are a significant factor hindering the healthy development of the industry. Among these, "white spot disease" caused by *Ichthyophthirius multifiliis* and "trichodina disease" caused by *Trichodina spp.* are particularly serious. Currently, traditional chemical drugs such as formaldehyde and copper sulfate are still widely used in aquaculture practices. Long-term use of these drugs can easily lead to problems such as parasite resistance, drug residues in aquatic products, and environmental pollution.

[0003] To this end, Chinese invention patent application number 201410492882.8 discloses an ectoparasite control agent for aquatic animals. The active ingredient of this preparation is eugenol, which has the advantages of low toxicity, non-toxicity to aquatic animals, and low residue.

[0004] However, both *Ichthyophthirius multifiliis* and *Chilodonella spp.* parasitize the skin and gills of fish. A common characteristic is their ability to induce the host to produce large amounts of mucus and to form dense body surface structures or cyst walls (such as the cysts of *Ichthyophthirius multifiliis*), collectively constituting an effective physical and biochemical barrier. Because eugenol or small molecule compounds have very weak penetration into this barrier, the drug cannot reach the target site in sufficient quantity, resulting in low insecticidal efficacy.

[0005] Meanwhile, eugenol or traditional small molecule compounds with antibacterial properties have extremely poor water solubility, making it difficult to form an effective concentration in water. They also have the disadvantages of being chemically unstable and easily degraded, resulting in very limited effectiveness when used directly as insecticides. Summary of the Invention

[0006] This invention provides a transmembrane drug delivery system and its preparation method, as well as its application. The transmembrane drug delivery system can penetrate the host's mucus and parasite barrier, thereby accurately delivering the effective medicinal ingredients to the target site and achieving the effect of killing fish parasites.

[0007] The objective of this invention is achieved through the following technical solution: A transmembrane drug delivery system comprises a polyphenol compound with antiparasitic activity, a drug carrier, and a penetration enhancer; wherein the mass ratio of the polyphenol compound to the penetration enhancer is 0.5-1:0.5-3.

[0008] Preferred polyphenolic compounds are curcumin and gallic acid, or a mixture thereof. Curcumin, gallic acid, and other plant-derived polyphenolic compounds are considered highly promising candidates for green veterinary drugs due to their diverse biological activities, including antibacterial, anti-inflammatory, and antiparasitic properties, as well as their good environmental compatibility.

[0009] Preferably, the penetration enhancer is at least one of carvone, geraniol, and borneol. Through the action of the penetration enhancer, the drug can penetrate the parasite barrier and be efficiently delivered to the parasitic target.

[0010] Preferably, the drug carrier includes a matrix, penetration enhancer A, and penetration enhancer B; The matrix is ​​oleic acid; The penetration aid A is selected from at least one of polyoxyethylene hydrogenated castor oil and Tween 80; The penetration enhancer B is selected from at least one of ethanol, methanol, and PEG400; The mass ratio of the matrix, penetration enhancer A, and penetration enhancer B is 2-8:4-12:1-6.

[0011] Preferably, penetration enhancer A is polyethylene hydrogenated castor oil, penetration enhancer B is ethanol, and the mass ratio of the matrix, penetration enhancer A and penetration enhancer B is 3:5:1.67.

[0012] This invention also discloses a method for preparing a transmembrane drug delivery system, the method comprising the following steps: S01. Add the polyphenol compound to the drug carrier and stir in a water bath at 50℃-70℃ until completely dissolved to obtain a drug-containing oil phase. S02. Mix penetration aid A, penetration aid B and penetration promoter, and stir evenly to obtain a mixed phase; S03. Under high-speed shearing conditions, the drug-containing oil phase and the mixed phase are thoroughly mixed to form a uniform and transparent premix. S04. Under continuous stirring, the premix is ​​slowly added to ultrapure water preheated to 35℃-40℃, and stirring is continued for more than 30 minutes to obtain a homogeneous and transparent transmembrane drug delivery system solution.

[0013] Preferably, in step S01, the water bath temperature is 60°C.

[0014] Preferably, in step S01, the conditions for high-speed shearing are: shearing at room temperature with a rotation speed of 8000-12000 rpm for 3-5 minutes.

[0015] Preferably, in step S04, the preheating temperature of the ultrapure water is 37°C.

[0016] The present invention also provides applications of transmembrane drug delivery systems, including the following (1) or (2); (1) As a drug that can kill both Ichthyophthirius multifiliis and Trichodina ectenes; (2) Prevention and control of fish parasites.

[0017] Compared with the prior art, the advantages or beneficial effects of the technical solution of this application include: 1. The transmembrane drug delivery system provided by this invention integrates a specific permeation enhancer into the drug delivery system. This enhancer works synergistically with the carrier complex to primarily exert a physical effect on disrupting biological barriers. This integrated drug delivery and permeation enhancement design specifically addresses the low penetration efficiency of green drugs in vitro, fundamentally differing from conventional technologies that only improve solubility or achieve sustained release. This drug delivery system is particularly suitable for killing fish parasites with dense body surface structures or cyst walls.

[0018] 2. The drug carrier provided by this invention can effectively load hydrophobic polyphenol drugs and simultaneously and significantly enhance their killing activity against Ichthyophthirius multifiliis and Chilodonella spp. Furthermore, the hydrophobic polyphenol drugs are encapsulated within the drug carrier, which increases the structural stability of the effective medicinal components and improves their solubility in water, thus facilitating delivery to the lesions on the fish and achieving the insecticidal effect. Attached Figure Description

[0019] Figure 1 This is an example of the permeability and in vitro insecticidal effect of Cur-DDS and curcumin raw materials on Ichthyophthirius multifiliis in Example 4 of the present invention. (A) shows a comparison of dose-response curves for in vitro killing of Ichthyophthirius multifiliis; (B) shows a comparison of fluorescence microscopy images evaluating the permeability of Cur-DDS to the cyst membrane of Ichthyophthirius multifiliis based on AO / PI staining.

[0020] Figure 2 This illustrates the insecticidal effect of Cur-DDS and Cur on Ichthyophthirius multifiliis on the surface of goldfish in Example 4 of this invention.

[0021] Figure 3 The images show H&E staining of pathological sections of liver tissue from goldfish after 21 days of subchronic exposure to different concentrations of Cur-DDS in Example 6 of this invention. In the images, (A) represents the control group; (B) represents the 2 mg / L group; (C) represents the 2.5 mg / L group; and (A) represents the 4 mg / L group.

[0022] Figure 4 This study describes the effects of different concentrations of Cur-DDS on hematological parameters of goldfish after 21 days of treatment, as described in Example 6 of this invention.

[0023] Figure 5 This study describes the effects of different concentrations of Cur-DDS on the biochemical indicators of goldfish serum after 21 days of treatment, as described in Example 6 of this invention. Detailed Implementation

[0024] The following detailed description of the embodiments of this application, in conjunction with the accompanying drawings, will provide a thorough understanding of how this application uses technical means to solve technical problems and achieve corresponding technical effects, enabling its implementation. The embodiments of this application and the various features within them can be combined with each other without conflict, and all resulting technical solutions are within the protection scope of this application.

[0025] Example 1: Construction of Drug Delivery System This transmembrane drug delivery system consists of three parts: a polyphenol compound, a drug carrier, and a penetration enhancer. The active ingredient is curcumin or gallic acid, and its dosage is 0.1% of the total system mass.

[0026] The drug carrier contains oleic acid as a matrix component, and a penetration enhancer composed of polyoxyethylene hydrogenated castor oil (RH40) and ethanol.

[0027] The penetration enhancer is carvone, geraniol, or borneol, and its dosage is 0.2% of the total system mass. The mass ratio of oleic acid, RH40, and anhydrous ethanol is 3:5:1.67. The remaining balance of the system is made up with ultrapure water.

[0028] The specific preparation process of this transmembrane drug delivery system is as follows: First, curcumin or gallic acid is added to oleic acid and stirred in a 60°C water bath until completely dissolved to obtain a drug-containing oil phase. Second, RH40, anhydrous ethanol, and carvone, geraniol, or borneol are mixed and stirred evenly to obtain a mixed phase. Subsequently, the drug-containing oil phase and the mixed phase are thoroughly mixed under high-speed shearing conditions (10,000 rpm for 4 min) to form a homogeneous and transparent premix. Finally, under continuous stirring, the premix is ​​slowly added to sufficient ultrapure water preheated to 37°C, and stirring is continued for more than 30 min to obtain a homogeneous and transparent curcumin or gallic acid-loaded system solution.

[0029] Example 2: Screening test of penetration enhancer This embodiment aims to screen for permeation enhancers that can effectively improve the ability of drug delivery systems to penetrate biological barriers.

[0030] (1) Franz diffusion tank transdermal test: Intact skin from the back of a healthy goldfish was taken and fixed in a Franz diffusion tank (release area 2.54 cm²). 2The receiving chamber had a volume of 7 mL. A base solution (containing only the carrier complex, not the drug) containing 1% of different candidate permeation enhancers (carvone, borneol, geraniol, salicylic acid, lecithin, menthol, stearic acid, and azone) was added to the supply tank. The receiving tank was filled with 34℃ physiological saline, magnetically stirred at 300 r / min, and kept at a constant temperature of (34±0.5)℃ in a water bath. 2 mL of the receiving solution was collected at 0, 1, 2, 3, 4, 5, 6, 7, 8, 10, 12, 24, 36, and 48 h, and replenished. After filtration through a 0.45 μm filter membrane, the cumulative permeation of curcumin (Q) was determined by HPLC. The cumulative permeation at 48 h (Q) was used as the basis for the calculation. 48 () is the main evaluation indicator.

[0031] (2) Agar diffusion test: A 2% mucin agarose gel containing fish mucus was prepared in a 6-well plate. The base solution of the drug delivery system containing different penetration enhancers (concentration of 50% or 100%) was added to the surface of the gel. After 24 h, the vertical penetration depth of the nanoemulsion in the gel was accurately measured.

[0032] (3) Experimental results: As shown in Table 1, the samples containing carvone, borneol, or geraniol had a greater penetration depth than all other candidate penetration enhancer groups and the control group. This indicates that carvone, borneol, and geraniol have excellent ability to disrupt the mucogel network structure and effectively reduce barrier resistance. Therefore, carvone, borneol, and geraniol were selected as the dedicated penetration enhancers for the drug delivery system of this invention.

[0033] Table 1. Screening results of different penetration enhancers

[0034] Example 3: Stability evaluation of drug delivery system (taking curcumin drug delivery system as an example) (1) Stability during storage: The curcumin drug delivery system (Cur-DDS) stock solution was stored at 4°C for 6 months. The appearance was observed regularly and key indicators were measured. The results are shown in Table 2. During storage, the sample remained clear and translucent without stratification or precipitation. After 6 months, the drug loading decreased slightly from 0.796 mg / mL to 0.702 mg / mL, the encapsulation efficiency decreased from 98.0% to 86.4%, and the average particle size and PDI did not change significantly, proving that the formulation had good stability within 6 months.

[0035] Table 2 Results of the Cur-DDS preservation stability test

[0036] (2) Stability of storage media: Cur-DDS was diluted 10 times with ultrapure water, hydrochloric acid solution with pH=1.2 and phosphate buffer with pH=6.8 (containing 0.5% SDS), and placed at 4℃ for 96 h. As shown in Table 3, the drug loading and encapsulation efficiency of Cur-DDS remained stable in different media with small variations, indicating that it has good acid and alkali resistance.

[0037] Table 3 Results of the stability evaluation test of Cur-DDS preservation medium

[0038] Example 4: Broad-spectrum antiparasitic application of curcumin-loaded systems 1. Cur-DDS's effect in killing Ichthyophthirius multifiliis (1) Permeation-enhancing assay based on AO / PI staining: 100 μL of worm solution containing approximately 100 cysts was added to a 96-well plate, followed by Cur-DDS to a final concentration of 0.5 mg / L. The plate was treated for 30 min. The worms were collected by centrifugation and stained according to the AO / PI cell viability assay kit instructions. After incubation in the dark for 5 min, the cells were observed under a fluorescence microscope. The results are as follows: Figure 1 As shown in Figure A: AO dye was uniformly distributed within the cysts in both the control group and the Cur-DDS group, indicating that it has good membrane permeability. The cysts in the Cur-DDS-treated group showed a large amount of strong red PI fluorescence (membrane damage), while the control group mainly showed green AO fluorescence (membrane integrity), which directly proves that Cur-DDS can effectively disrupt the integrity of the Ichthyophthirius multifiliis cyst membrane and promote drug penetration.

[0039] (2) In vitro killing test for Ichthyophthirius multifiliis: Goldfish with typical white spot disease symptoms were selected, and trophozoites were collected by rinsing with ultrapure water and incubated overnight in a constant temperature incubator at 22℃ to hatch predators. The predators were collected by centrifugation for the experiment. Goldfish naturally infected with Ichthyophthirius multifiliis were randomly selected, and caudal fin tissue blocks containing about 30-50 Ichthyophthirius multifiliis were cut and placed in 24-well plates. 1 mL of different concentrations (0, 0.5, 1, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10 mg / L) of Cur-DDS, Cur-DDS drug excipient (without curcumin, PE), or curcumin solution (Cur) were added to each well, with 3 replicates per group. Under constant temperature conditions, after 4 h, the number of dead parasites was observed and recorded under an inverted microscope (death criteria: complete cessation of movement and cyst rupture), and the killing rate and EC50 were calculated. The results are as follows: Figure 1As shown in Figure B, Cur-DDS achieved a 100% kill rate against Ichthyophthirius multifiliis (white spot disease) at a concentration of 2 mg / L over 4 hours, compared to 60% at 1 mg / L and 34.6% at 0.8 mg / L, with a 4-hour EC50 of 0.96 mg / L. In contrast, the 100% insecticidal concentration of Cur solution was 8 mg / L, four times that of Cur-DDS. The excipient PE group showed no insecticidal activity, indicating that the excipient had no inhibitory effect on Ichthyophthirius multifiliis.

[0040] (3) Cur-DDS treatment trial for Ichthyophthirius multifiliis in goldfish Goldfish exhibiting typical symptoms of Ichthyophthirius multifiliis infection (dense white spots, sluggish swimming, and decreased appetite) were selected and temporarily housed in well-aerated aquariums at a water temperature of (20±1)℃. The experiment was conducted in 50 L plastic tanks, with 10 infected fish randomly placed in each tank. Based on the results of in vitro parasite control, different concentrations of Cur-DDS or Cur (0, 2, 4, 6, 8, 10 mg / L, calculated as curcumin concentration) were set, with three replicates per group. During the experiment, 50% of the water was changed daily, and the corresponding concentration was re-added, continuing the treatment for 96 hours. Abnormal behavior and mortality were observed and recorded daily, and dead fish were removed promptly. After the experiment, three surviving fish were randomly selected from each tank, and mucus and epidermal tissue were gently scraped from their bodies, placed on a glass slide, and observed under a microscope in at least three fields of view. The number of surviving and dead Ichthyophthirius multifiliis was counted, and the parasite control rate and EC were calculated. 50 The result is as follows Figure 2 As shown, Cur-DDS achieved a 100% kill rate against Ichthyophthirius multifiliis at a concentration of 2 mg / L, and the white spots on the fish completely disappeared after 96 h of treatment. The 100% kill concentration of Cur solution was 10 mg / L, indicating that Cur-DDS can enhance the killing activity against Ichthyophthirius multifiliis.

[0041] 2. The killing effect of Cur-DDS on *Ichthyophthirius multifiliis* (1) Cur-DDS in vitro killing test for *Ichthyophthirius multifiliis*: Parasites were scraped from the gills and body surface of goldfish infected with *Ichthyophthirius multifiliis*, and a suspension of live parasites was obtained after washing with sterile PBS. 1 mL of the suspension containing approximately 30-50 *Ichthyophthirius multifiliis* was added to each well of a 24-well plate. Then, 1 mL of different concentrations of Cur-CNE, Cur-DDS excipients (excluding curcumin, PE), or curcumin solution (Cur) were added to achieve final concentrations of 4, 2, 1, 0.5, 0.25, 0.13, 0.06, 0.03, and 0 mg / L (based on curcumin concentration), with three replicates per group. The plates were kept at 22℃ for 4 h and then observed under an inverted microscope. The mortality rate and 4-hour EC50 were calculated based on the complete cessation of ciliary movement, parasite deformation, or disintegration. 50The results are shown in Table 4: Cur-DDS exhibited highly efficient killing activity against *Ichthyophthirius multifiliis*, with a 100% kill concentration and EC50 at 4 h. 50 The concentrations were 0.06 and 0.025 mg / L, respectively, significantly better than the Cur API group (100% kill concentration and EC50). 50 The concentrations were 1 and 0.41 mg / L, respectively. All concentrations of Cur-DDS excipients showed no killing activity against *Chilodonella esculenta*, indicating that the excipients had no inhibitory effect on *Chilodonella esculenta*.

[0042] Table 4. In vitro killing effects of different drugs on *Ichthyophthirius multifiliis*

[0043] (2) Treatment experiment of Cur-DDS for goldfish with chilodonella infection: Goldfish with a large number of chilodonella parasites on their gills and body surface confirmed by microscopic examination were selected and randomly divided into Cur-DDS or Cur experimental groups. Based on the in vitro parasite killing results, different concentrations of Cur-DDS or Cur were set (8, 4, 2, 1, 0.5, 0.25, 0.13, 0.06, 0 mg / L, calculated as curcumin concentration), with 3 replicates in each group and 10 fish in each replicate. The experimental group was treated with a bath, while the negative control group was treated with aerated water. 50% of the water was changed and the drug was added again every day for 96 hours. The mucus on the fish body surface and the respiratory status were observed every day. 96 hours after treatment, 3 fish were randomly selected from each group, and gill filaments were taken to make water-soaked slides. The average number of parasites in at least 3 fields of view were counted under a microscope to calculate the parasite killing rate.

[0044] The results showed that after 96 hours, the negative control group (i.e., the 0 mg / L group) exhibited increased mucus secretion and a persistently high number of parasites on the gills. In contrast, the goldfish treated with Cur-DDS at concentrations above 0.25 mg / L showed reduced mucus secretion on the gills and a significant decrease in the number of parasites detected under microscopic examination. As shown in Table 5, the 100% kill concentration of the Cur-DDS treatment group was 0.5 mg / L, significantly higher than that of the Cur solution group (4 mg / L), indicating that Cur-DDS has a significant therapeutic effect on chilodonella infection in goldfish.

[0045] Table 5. Killing effects of different drugs on *Ichthyophthirius multifiliis* on the surface of goldfish.

[0046] Example 5: Broad-spectrum antiparasitic application of the gallic acid-loaded system Referring to the composition and proportion of the drug carrier in Example 1, the active ingredient was replaced with gallic acid, and the penetration enhancer was replaced with carvone, with an addition amount of 1% of the total mass. The gallic acid-loaded system was constructed using the same process. The in vitro insecticidal activity of this system against *Ichthyophthirius multifiliis* and *Chilodonella ectenes* was evaluated. The same in vitro evaluation method as in Example 4 was used. The results are shown in Table 6: EC50 of gallic acid raw material against *Ichthyophthirius multifiliis* and *Chilodonella ectenes*. 50 The effects were 5.5 times and 8.3 times greater than those of the gallic acid-loaded system, respectively. This embodiment further verifies that the drug delivery system of the present invention has good compatibility with different active ingredients (gallic acid) and different penetration enhancers (carvone). The system can also universally and significantly enhance the killing activity of the loaded drug against a variety of parasites, which further verifies the potential of the drug delivery system as a universal synergistic platform.

[0047] Table 6. In vitro killing effect of gallic acid-loaded system on *Ichthyophthirius multifiliis*

[0048] Example 6: Safety Evaluation (Taking Curcumin-loaded System as an Example) 1. Acute toxicity test of Cur-DDS on goldfish Healthy goldfish (15.2 g ± 1.8 g) were randomly assigned to glass tanks containing 40 L of aerated water, with 20 fish per tank. A Cur-DDS concentration gradient was established: 0 (control), 2, 4, 6, 8, and 10 mg / L, with three replicates per group. Exposure lasted for 96 h. Mortality was observed and recorded daily (death was defined as cessation of gill movement and lack of response to mild mechanical stimulation), and dead fish were removed promptly. At the end of the experiment, the cumulative mortality rate over 96 h was calculated. The median lethal concentration (LC50) was calculated using probability analysis. The results showed that no goldfish died within 96 h when the Cur-DDS concentration was less than 6 mg / L; deaths occurred in the 8 mg / L and 10 mg / L concentration groups. The 96-h LC50 of Cur-DDS for goldfish was calculated. 50 The concentration was 8.7 mg / L, which is much higher than the 100% effective therapeutic concentration for Ichthyophthirius multifiliis (2 mg / L), indicating that its safety window is relatively wide.

[0049] 2. Subchronic toxicity test of Cur-DDS on goldfish (1) Experimental Design: Referring to the "Guiding Principles for Safety Evaluation Research of Target Animals in Aquatic Drug Use", a control group (0 mg / L) and three Cur-DDS exposure groups were set up: 2 mg / L (1 times the effective concentration), 2.5 mg / L (1.25 times), and 4 mg / L (2 times). Healthy goldfish were randomly divided into 4 groups after temporary holding, with 3 replicates in each group and 10 fish per replicate. They were housed in 40 L of aerated tap water at a temperature of (20±1.5)℃ for 21 days. The water was changed and the drug was re-added every 48 hours. Mortality was recorded daily.

[0050] (2) Histopathological observation: On day 21, three goldfish were randomly selected from each group. Liver tissue was taken, fixed with 4% paraformaldehyde, dehydrated with a routine gradient of ethanol, cleared with xylene, and embedded in paraffin. The section thickness was 4–6 μm. After H&E staining, the sections were observed and photographed under an optical microscope. The results showed that in the 21-day subchronic toxicity test, no goldfish in the Cur-DDS treatment groups (2, 2.5, and 4 mg / L) died and their vitality was normal. Histopathological sections of the liver tissue showed that the liver tissue structure of the goldfish in the Cur-DDS treatment groups was intact, the hepatocyte cords were neatly arranged, and the morphology was normal. Compared with the control group, no obvious pathological damage such as edema, fatty degeneration, necrosis, or inflammatory cell infiltration was observed. The results are as follows. Figure 3 As shown.

[0051] (3) Hematological and serum biochemical index detection: At the end of the experiment, blood was collected from the tail vein of the fish using a syringe rinsed with heparin sodium. A portion of the whole blood was used to determine the white blood cell count (WBC), red blood cell count (RBC), hemoglobin concentration (HGB), and platelet count (PLT) using an automated hematology analyzer. The other portion of the blood was centrifuged to separate serum, and the alkaline phosphatase (AKP), alanine aminotransferase (ALT), creatinine (SCR), and total cholesterol (T-CHO) levels were measured using a commercially available reagent kit on an enzyme-linked immunosorbent assay (ELISA) reader. The results showed that there were no statistically significant differences in any hematological and serum biochemical indexes between the 2 mg / L and 2.5 mg / L treatment groups and the control group (P>0.05); the WBC and HGB levels in the 4 mg / L treatment group were slightly decreased, and the RBC level was slightly increased, but all indicators were within the normal physiological fluctuation range. The results are as follows. Figure 4 As shown in the figure. Serum biochemical indicators (AKP, ALT, SCR, T-CHO) did not change significantly among the treatment groups, as shown in the figure. Figure 5 As shown, this indicates that under 21 days of subchronic exposure, Cur-DDS did not cause significant hematologic toxicity or liver and kidney damage in goldfish.

[0052] The embodiments described above are some, but not all, of the embodiments of the present invention. The detailed description of the embodiments of the present invention is not intended to limit the claimed invention to a tedious level, but merely to illustrate selected embodiments of the invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without inventive effort are within the scope of protection of the present invention.

Claims

1. A transmembrane drug delivery system, characterized in that, It is composed of a polyphenol compound with antiparasitic activity, a drug carrier, and a penetration enhancer; wherein the mass ratio of the polyphenol compound to the penetration enhancer is 0.5-1:0.5-3.

2. The transmembrane drug delivery system according to claim 1, characterized in that, The polyphenolic compounds are curcumin and gallic acid, or a mixture thereof.

3. The transmembrane drug delivery system according to claim 1, characterized in that, The penetration enhancer is at least one of carvone, geraniol and borneol.

4. The transmembrane drug delivery system according to claim 1, characterized in that, Drug carriers include a matrix, penetration enhancer A, and penetration enhancer B; The matrix is ​​oleic acid; The penetration enhancer A is selected from at least one of polyoxyethylene hydrogenated castor oil and Tween 80; The penetration enhancer B is selected from at least one of ethanol, methanol, and PEG400; The mass ratio of the matrix, penetration enhancer A, and penetration enhancer B is 2-8:4-12:1-6.

5. The transmembrane drug delivery system according to claim 1, characterized in that, Penetration aid A is polyoxyethylene hydrogenated castor oil, and penetration aid B is ethanol. The mass ratio of the matrix, penetration aid A, and penetration aid B is 3:5:1.

67.

6. A method for preparing a transmembrane drug delivery system as described in any one of claims 1-5, characterized in that, Includes the following steps: S01. Add the polyphenol compound to the drug carrier and stir in a water bath at 50℃-70℃ until completely dissolved to obtain a drug-containing oil phase. S02. Mix penetration aid A, penetration aid B and penetration promoter, and stir evenly to obtain a mixed phase; S03. Under high-speed shearing conditions, the drug-containing oil phase and the mixed phase are thoroughly mixed to form a uniform and transparent premix. S04. Under continuous stirring, the premix is ​​slowly added to ultrapure water preheated to 35℃-40℃, and stirring is continued for more than 30 minutes to obtain a homogeneous and transparent transmembrane drug delivery system solution.

7. The preparation method according to claim 6, characterized in that, In step S01, the water bath temperature is 60℃.

8. The preparation method according to claim 6, characterized in that, In step S01, the conditions for high-speed shearing are: at room temperature, shearing at a rotation speed of 8000-12000 rpm for 3-5 minutes.

9. The preparation method according to claim 6, characterized in that, In step S04, the preheating temperature of the ultrapure water is 37°C.

10. The application of the transmembrane drug delivery system according to any one of claims 1-5 and the transmembrane drug delivery system prepared by any one of claims 6-9, characterized in that, Including the following (1) or (2); (1) As a drug that can kill both Ichthyophthirius multifiliis and Trichodina ectenes; (2) Prevention and control of fish parasites.

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