Hyperoside mixed micelle, preparation method therefor, and use thereof

Abstract

Disclosed in the present invention are a hyperoside mixed micelle, a preparation method therefor, and use thereof. The hyperoside polymer mixed micelle comprises: hyperoside, TPGS, and F68. The polymers TPGS and F68 are used as carriers for the mixed micelle to encapsulate the hyperoside therein. The hyperoside mixed micelle of the present invention can effectively increase the solubility of the hyperoside, thereby improving its oral bioavailability, reducing adverse drug reactions, and enhancing the therapeutic efficacy against ulcerative colitis.

Description

A kind of hyperoside mixed micelle and its preparation method and application Technical Field

[0001] The invention belongs to the field of pharmaceutical preparations, and particularly relates to a polymer mixed micelle containing hyperoside, a poorly soluble anti-inflammatory drug, for treating ulcerative colitis and a preparation method thereof. Background Art

[0002] Ulcerative colitis is a chronic, persistent, relapsing and remitting inflammatory disease of the colonic mucosa. Its incidence is increasing worldwide. It can develop at any age, but most commonly occurs in young adults. In my country, the peak age of onset is 20 to 49 years. Due to the relatively high incidence of relapsing ulcerative colitis, patients often require long-term medication to control disease progression. Current treatment options for ulcerative colitis are limited to aminosalicylates, sulfasalazine, antibiotics, steroids, and immunosuppressants, but these treatments lack efficacy and are associated with numerous side effects. Investigational biotherapeutics (anti-tumor necrosis factor antibodies, interferon) often generate anti-drug antibodies during treatment, making long-term use difficult. Therefore, there is an urgent need to explore new avenues for the treatment of colitis with novel mechanisms of action. Flavonoids, particularly those with anti-inflammatory and antioxidant pharmacological activities, may represent potential therapeutic agents for the prevention and treatment of intestinal diseases.

[0003] Hyperoside (quercetin-3-Od-galactoside, HYP) is a naturally occurring flavonoid compound belonging to the flavanol glycoside family. It is widely found in medicinal herbs such as licorice, hibiscus flower, and Apocynum venetum leaf and exhibits excellent anti-inflammatory and antioxidant activities. Studies have also demonstrated that hyperoside has certain preventive and therapeutic effects on ulcerative colitis. Literature reports suggest that hyperoside ameliorates DSS-induced colitis through MKRN1-mediated regulation of PPARγ signaling and Th17 / Treg balance. However, the poor water solubility and low oral bioavailability of hyperoside limit its further application in ulcerative colitis. Therefore, the design and preparation of a safe and effective drug delivery system is crucial to enhance the colitis-preventive and therapeutic effects of HYP.

[0004] Summary of the Invention

[0005] The purpose of the present invention is to provide a hyperoside mixed micelle and a preparation method and application thereof in view of the defect of poor water solubility of hyperoside.

[0006] In order to achieve the above object, the present invention adopts the following technical solution: a hyperoside polymer mixed micelle, comprising: hyperoside, TPGS and F68, wherein the polymer TPGS and F68 are used as carriers of the mixed micelle, and hyperoside is encapsulated therein.

[0007] Furthermore, the mass ratio of hyperoside, TPGS and F68 is 1:2:6-1:6:6, more preferably 1:4:3-1:4:7, and most preferably 1:(2-6):6.

[0008] Furthermore, the particle size of the polymer mixed micelles is 20-40 nm, the PDI value is 0.150-0.200, the potential value is -18--25 mV, the encapsulation efficiency is 85%-95%, and the drug loading is 7%-11%.

[0009] A method for preparing the above-mentioned hyperoside polymer mixed micelles comprises the following steps:

[0010] (1) Dissolve hyperoside, TPGS, and F68 in an organic solvent and ultrasonicate to form a uniform mixed solution;

[0011] (2) controlling the temperature of the rotary evaporator to 45°C to evaporate the organic solvent of the mixed solution to obtain a hyperoside film with TPGS and F68 as carriers;

[0012] (3) The hyperoside film with TPGS and F68 as carriers was hydrated in double distilled water to obtain a hyperoside polymer mixed micelle preparation.

[0013] Furthermore, the organic solvent in step (1) is methanol.

[0014] A use of the hyperoside polymer mixed micelles in the preparation of drugs for treating ulcerative colitis.

[0015] Furthermore, the daily dosage of the hyperoside polymer mixed micelles is 100 mg / kg to 50 mg / kg.

[0016] By measuring the in vitro cumulative release of the prepared hyperoside polymer mixed micelles and hyperoside in four media (double distilled water, Ph1.2 hydrochloric acid solution, Ph6.8 and Ph7.4 buffer solutions), it was shown that the micelles of the present invention can increase the solubility of hyperoside.

[0017] Pharmacokinetic experiments on rats and in vivo imaging experiments confirmed that the hyperoside polymer mixed micelles prepared in the present invention can prolong the drug action time and improve the oral bioavailability of hyperoside.

[0018] A mouse enteritis model was established by inducing DSS aqueous solution, and hyperoside and hyperoside micelles were administered orally for intervention. The anti-mouse enteritis efficacy of hyperoside and hyperoside micelles was evaluated by daily (Disease Activity Index) DAI score, body weight changes, colon length changes and spleen index, cytokine level analysis, changes in MPO activity in colon tissue, and histopathological evaluation. The results showed that hyperoside micelles can enhance the efficacy of hyperoside against ulcerative colitis.

[0019] Currently, nanoparticle drug delivery systems are widely used in the treatment of ulcerative colitis, showing great promise in improving pharmacokinetics, bioavailability, and drug solubility. Nanoparticles are more readily absorbed by target cells in inflamed intestinal areas due to their larger surface area, making them more likely to be retained in the intestine, resulting in a longer residence time in inflamed areas. This property also prevents nanoparticle loss due to diarrhea. Compared with traditional drug delivery systems, nanoparticle drug delivery systems can reduce the required effective drug dose, thereby reducing drug side effects. Polymeric micelles, as one of the nanoparticle drug delivery systems, have long been a preferred carrier due to their advantages of prolonging drug residence time in the circulation, improving the solubility of hydrophobic drugs in aqueous phases, and reducing drug toxicity and side effects. Hyperoside-encapsulated polymer mixed micelles have emerged as a promising nanoformulation due to their small particle size, strong targeting, increased drug solubility, and improved oral bioavailability.

[0020] The beneficial effects of the present invention are as follows: the polymer mixed micelles in the present invention use TPGS and F68 as drug carriers. TPGS is pegylated vitamin E, which has good biocompatibility, can solubilize, can enable drugs to achieve long-term circulation in the body and is conducive to cellular uptake, etc. F68 is an FDA-approved carrier material for the preparation of injections, which has low toxicity and good biocompatibility. F68 has been proven to have the effect of inhibiting cytochrome P450 enzymes and their isoforms, and can change the metabolism and elimination of drugs.

[0021] The polymer mixed micelles of the present invention can increase the solubility of drugs, improve the oral bioavailability of drugs, reduce adverse drug reactions and improve therapeutic effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] FIG1 is an electron microscope image and a particle size distribution diagram of the hyperoside-loaded polymer mixed micelles (HYP-M) in the embodiment.

[0023] FIG2 is an in vitro cumulative release curve of hyperoside and HYP-M in Example.

[0024] FIG3 is a diagram showing the drug concentration distribution of hyperoside and HYP-M in serum of the embodiment.

[0025] FIG4 is a mouse in vivo imaging diagram of the polymer mixed micelles of the embodiment

[0026] FIG5 is a graph showing changes in body weight of mice in each group during the treatment of DSS-induced ulcerative colitis using hyperoside and HYP-M in the example.

[0027] FIG6 is a graph showing the DAI scores of mice in each group during the treatment of DSS-induced ulcerative colitis using hyperoside and HYP-M in the example.

[0028] FIG7 is a graph showing the changes in colon length of mice in each group after treatment of DSS-induced ulcerative colitis with hyperoside and HYP-M in the example.

[0029] FIG8 is a graph showing changes in spleen index of mice in each group after treatment of DSS-induced ulcerative colitis with hyperoside and HYP-M in the example.

[0030] FIG9 is a graph showing the serum cytokine TNF-α, IL-6, and IL-1β levels in mice in each group after treatment of DSS-induced ulcerative colitis with hyperoside and HYP-M in the example.

[0031] FIG10 is a graph showing the MPO activity in the colon tissues of mice in each group after DSS-induced ulcerative colitis was treated with hyperoside and HYP-M in the Example.

[0032] FIG11 is a diagram of tissue sections of mice in each group treated with hyperoside and HYP-M for DSS-induced ulcerative colitis in the example. DETAILED DESCRIPTION

[0033] In order to enable those skilled in the art to better understand the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments in the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts should fall within the scope of protection of this application.

[0034] It should be noted that the terms "including" and "having" and any variations thereof in the specification and claims of this application and the above-mentioned drawings are intended to cover non-exclusive inclusions. For example, a process, method, system, product or apparatus that includes a series of steps or units is not necessarily limited to those steps or units clearly listed, but may include other steps or units that are not clearly listed or are inherent to these processes, methods, products or apparatuses.

[0035] Example 1: Preparation and stability testing of HYP-M

[0036] 1. Preparation of HYP-M

[0037] HYP-M was prepared using a thin film dispersion method. 5 mg of hyperoside, 20 mg of GPTS, and 30 mg of F68 were precisely weighed into a 100 mL round-bottom flask. 10 mL of methanol was added and thoroughly dissolved by sonication for 15 minutes. The methanol was then evaporated to dryness using a rotary evaporator at 45°C. A uniform thin film formed on the inner wall of the flask. HYP-M was then hydrated with 10 mL of double-distilled water to obtain HYP-M. Figure 1 shows that HYP-M is spherical, monodispersed, and has a relatively uniform particle size distribution. The average particle size of HYP-M is 21.48 ± 1.37 nm, the PDI value is 0.178 ± 0.013, the potential value is -20 ± 0.48 mV, the encapsulation efficiency is 89.59 ± 2.03%, and the drug loading is 8.18 ± 0.18%.

[0038] 2. Stability test of HYP-M

[0039] The micellar films were stored at 4°C and hydrated after 7, 14, and 30 days. The micelle particle size, PDI, and potential were measured to investigate the stability of the HYP-M films under 4°C storage conditions. The P values ​​for the potential and PDI values ​​of the first and other groups were calculated, and P values ​​> 0.05 were found. The particle size and PDI values ​​of the hyperoside micelles remained unchanged after 30 days of storage. While the potential of the micelle preparation showed a slight decrease after hydration, its absolute value remained greater than 10 mV. No drug precipitation was observed after hydration, indicating that the hyperoside micelle preparation films exhibited good stability under these storage conditions. The results are listed in Table 1.

[0040] Table 1

[0041] Example 2: Determination of HYP-M in vitro cumulative release

[0042] Hyperoside solutions and hyperoside polymer mixed micelle solutions of equal concentration were prepared. 1 mL of each solution was added to a dialysis bag, which was then placed in 100 mL of four different media (double-distilled water, pH 1.2 hydrochloric acid solution, pH 6.8, and pH 7.4 buffer solutions) and shaken at 37°C. 1 mL of the sample was taken at intervals of 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, 8 h, 10 h, 12 h, and 24 h, and 1 mL of the corresponding media was added to maintain the dialysis bag in 100 mL of the media. 200 μL of the original 1 mL solution was removed and placed in a 1.5 mL EP tube. 200 μL of chromatographic methanol was added and vortexed to mix thoroughly. After centrifugation at 10,000 rpm for 10 min, 200 μL was taken and analyzed by high-performance liquid chromatography. The results are shown in Figure 2. In the four media, the in vitro cumulative release of the hyperoside preparation group was higher than that of the raw material drug group, that is, the solubility of the drug-loaded micelles in the preparation group was higher than that of the raw material drug, indicating that the solubility of hyperoside prepared into a nanoformulation using the micelles designed in this prescription as a carrier increased its solubility.

[0043] Example 3: Determination of Hyperoside and HYP-M Blood Concentrations in Rats

[0044] Before the experiment, 10 male SD rats were housed under good environmental conditions for 3 days and randomly divided into a hyperoside (HYP) group and a HYP-M group (n=5). All rats were fasted for 12 hours before administration and had free access to water. The HYP and HYP-M groups were orally administered with a hyperoside suspension and a HYP-M solution (single dose 300 mg / kg), respectively. After oral administration, approximately 0.5 mL of blood was collected by puncturing the retroorbital venous plexus at 0.085, 0.25, 0.5, 0.75, 1, 1.5, 2, 3, 4, 6, 8, 10, 12, and 24 hours. Blood samples were heated in a 37°C water bath for 20-30 minutes and centrifuged at 3700 rpm for 10 minutes to obtain 100 μL of serum. 50 μL of astragaloside IV methanol solution (6 μg / ml, internal standard) was vortexed with 100 μL of rat serum sample for 1 min, and then 800 μL of methanol was added to extract HYP in the plasma, and vortexed again for 1 min, and the methanol layer was collected. The collected methanol layer was dried under nitrogen at 37°C, and the remaining residue was dissolved in 100 μL of methanol again, vortexed for 2 min, centrifuged at 12000 rpm for 10 min, and 20 μL of supernatant was analyzed by high performance liquid chromatography system. The pharmacokinetic parameters of HYP and HYP-M were calculated by BAPP 2.3 software, and the results are shown in Table 2. The blood concentration-time distribution curves of HYP and HYP-M are shown in Figure 3. There are obvious differences in the blood concentration-time curves between the HYP group and the HYP-M group, and the blood concentration of the HYP-M group at each time point is higher than that of the HYP group. After oral administration, the blood concentration of the HYP-M group increased rapidly, and the highest concentration (C max ) was 1.41±0.06μg / mL, while C max The area under the plasma concentration-time curve (AUC 0-24h ) was 10.30±0.58h.μg / mL, while the AUC 0-24h The relative oral bioavailability of the HYP-M group was only 2.48±0.21μg / mL; compared with the HYP group, the relative oral bioavailability of the HYP-M group increased by 24%, and the oral bioavailability of the HYP-M group was about 5 times that of the HYP group. 1 / 2 ) than the HYP group (t 1 / 2 :9.31±0.96) was prolonged by 0.69h. HYP-M can prolong the circulation time of HYP in rats, promote the oral absorption of HYP, and thus increase the medicinal concentration and oral bioavailability of HYP.

[0045] Table 2

[0046] Example 4: In vivo imaging of mice

[0047] Fluorescein isothiocyanate was prepared into a 100 μg / mL ethanol solution, and micelles loaded with fluorescein isothiocyanate were prepared according to the micelle preparation method of Example 1.

[0048] Sixteen mice were randomly divided into four groups (n=4) and gavaged with 300 mg / kg of fluorescein isothiocyanate micelles. The mice were killed 1, 2, 4, and 6 hours after administration, and the stomach and intestinal tissues were removed for in vivo imaging. As shown in Figure 4, at 1 hour, the micelles were mainly retained in the stomach of the mice; after 2 hours, they were mainly in the stomach and duodenum; after 4 hours, micelles still remained in the stomach, and the micelles had moved to the lower part of the intestine, with fluorescent substances also distributed in the jejunum of the intestine; after 6 hours, the micelles were still in the stomach, and the fluorescent substances were mainly distributed in the jejunum and cecum. The in vivo imaging results show that the fluorescent preparation can be well retained in the intestine of the mice, and a large amount of fluorescent preparation is still retained in the jejunum and cecum at 6 hours, indicating that the micelles prepared by the present invention can greatly improve the absorption time of the drug and increase the oral bioavailability of the drug.

[0049] Example 5: Study on the intervention effect of HYP-M on DSS-induced ulcerative colitis

[0050] 1. Establishment of a mouse model of ulcerative colitis

[0051] Forty C57BL / 6 mice were placed in a standard laboratory environment and fed normally for one week to acclimate to the environment. The C57BL / 6 mice were randomly divided into 8 groups (n=5): a blank control group (N, free access to distilled water), a DSS model group (M, free access to 3.0% DSS aqueous solution), a positive control group (P, sulfasalazine tablets 500 mg / kg), a high-dose HYP group (HF-HYP, 100 mg / kg), a low-dose HYP group (LF-HYP, 50 mg / kg), a high-dose HYP-M group (H-HYP-M, 100 mg / kg), a low-dose HYP-M group (L-HYP-M, 50 mg / kg), and a blank micelle group (BP, 100 mg / kg). Except for the blank control group, mice in all other groups were allowed to drink 3.0% DSS aqueous solution daily. All other groups, except the blank control and DSS model groups, were given the corresponding drugs by gavage once daily for 8 consecutive days.

[0052] 2. Daily Disease Activity Index (DAI) score

[0053] The disease activity index (DAI) is a comprehensive score based on the patient's percentage of weight loss, stool consistency, and stool blood. The DAI value is calculated by dividing the total score of these three results by 3. Every morning at the same time, mice were observed for weight change (compared to their initial weight), stool consistency (whether diarrhea was present), and the presence of blood in the stool (whether occult blood or gross blood). The DAI value was then calculated to evaluate the efficacy of hyperoside in inhibiting enteritis in mice. The weight change results for mice are shown in Figure 5, and the DAI score results are shown in Figure 6. The specific scoring criteria are shown in Table 3. The method for determining occult blood in mouse feces is as follows: 1.0 g of o-tolidine was weighed and dissolved in a mixture of 50 mL each of glacial acetic acid and anhydrous ethanol to obtain 10.0 g / L o-tolidine. The solution was stored at 4°C in the dark until needed. 10 mL of H₂O₂ was diluted to 100 mL with ultrapure sterile water to obtain a 3.0% H₂O₂ solution, which was prepared daily for immediate use. At the same time every morning, use a sterile cotton swab to smear a small amount of mouse feces onto a clean glass slide. Add 2-3 drops of 10.0g / L o-tolidine, followed by several drops of 3.0% hydrogen peroxide, and observe the color change of the feces. Interpretation of occult blood results: Negative: No color change within 2 minutes; Weakly positive: From light blue to blue after 10 seconds; Positive: Initially appears light blue-brown, gradually turning distinct blue-brown; Strongly positive: Immediately turns blue-brown.

[0054] Table 3

[0055] As shown in Figure 5, the weight of mice in the blank control group continued to increase over time, while the model group mice that drank DSS lost approximately 16.3% of their weight after 8 days. This result is consistent with the clinical symptom of weight loss in ulcerative colitis and also confirms that the ulcerative colitis mouse model has been successfully established. The weight loss trend in the blank formulation group was almost identical to that in the model group, indicating that the TPGS and F68 in the formulation had no effect on the weight loss symptoms of ulcerative colitis mice. The figure clearly shows that oral administration of HYP and HYP-M effectively alleviated DSS-induced weight loss in mice, and HPY-M was more effective than HYP in alleviating DSS-induced weight loss in mice (P<0.001). As shown in Figure 6, the DAI index increased over time in each group. In the DSS model group, the DAI index was significantly higher than that in the drug-treated and normal groups (P<0.05), indicating that the ulcerative colitis mouse model was successfully established. The DAI index in the blank formulation group showed a similar increase as in the model group, indicating that the excipients used in the formulation had little effect on colonic inflammation. Compared with the HYP group, the DAI index of the HYP-M group was lower, indicating that the therapeutic effect of HYP-M was better than that of HYP.

[0056] 3. Determination of colon length and spleen index of each group of experimental mice

[0057] Eight days after inflammation, the experimental mice were sacrificed, and the abdominal cavity was quickly opened. Colon, kidney, and spleen tissues were collected 1 cm from the anus. The mouse colon length and spleen weight were measured, and the spleen index was calculated: spleen index (%) = spleen weight / mouse body weight × 100%. The experimental results are shown in Figures 7 and 8. As shown in Figure 7, the colon length of mice with DSS-induced ulcerative colitis was reduced compared to that of normal mice. The colon length of the model group and the blank formulation group was significantly shortened, and the shortening trend was similar. The excipients in the formulation had no effect on the shortening of the colon length of mice with enteritis. The shortening of the colon length of mice treated with HYP and HYP-M was less than that of the model group. Compared with HYP, HPY-M was more effective in alleviating DSS-induced shortening of the colon length of mice (P < 0.05). As shown in Figure 8, compared with the DSS model group, the spleen index of the drug-treated groups improved significantly, and the spleen index of the HYP-M group was smaller than that of the HYP group, indicating that HYP-M can effectively reduce side effects and enhance the therapeutic effect on immune organs. Furthermore, the therapeutic effect of the high-dose group was better than that of the low-dose group. Furthermore, the results showed that the colon length and spleen index of the blank micelle group were similar to those of the DSS model group (P>0.05), indicating that TPGS and F68 have no effect on the efficacy of ulcerative colitis.

[0058] 4. Determination of MPO activity in mouse colon tissue and serum cytokine TNF-α, IL-6, and IL-1β levels

[0059] Enzyme-linked immunosorbent assay (ELISA) kits were used to measure TNF-α, IL-6, and IL-1β levels in mouse serum, with absorbance read at 450 nm to assess the severity of inflammation. The experimental results, shown in Figure 9, show that the release of various early inflammatory factors was observed in the DSS-induced ulcerative colitis mouse model. Compared with the model group, the expression levels of TNF-α, IL-6, and IL-1β in colonic tissue in the HYP, HYP-M, and positive control groups were significantly reduced (P < 0.05). Compared with the HYP group, the expression levels of TNF-α, IL-6, and IL-1β in colonic tissue in the HYP-M group were even lower (P < 0.05), indicating that HYP-M can enhance the anti-inflammatory effect of HYP. Furthermore, the expression levels of TNF-α, IL-6, and IL-1β in colonic tissue in the blank micelle group were similar to those in the model group, with no significant differences (P > 0.05), indicating that TPGS and F68 had no significant effect on inflammation.

[0060] Colonic tissue MPO activity was determined strictly according to the kit instructions: 1) Accurately weigh colon tissue and prepare a 5.0% colon tissue homogenate using Reagent 2 as the homogenization medium at a weight-to-volume ratio of 1:19. 2) Take 0.9 mL of the prepared 5.0% colon tissue homogenate and add 0.1 mL of Reagent 3. Mix thoroughly and incubate in a 37°C water bath for 15 minutes. 3) To the control tube, add 3 mL of distilled water, 0.2 mL of sample, and 0.2 mL of Reagent 4 in that order; to the assay tube, add 3 mL of colorimetric reagent, 0.2 mL of sample, and 0.2 mL of Reagent 4 in that order. 4) Mix the control tube and the assay tube, incubate again in a 37°C water bath for 30 minutes, and then add 0.05 mL of Reagent 7 to each tube. 5) Mix the control tube with Reagent 7 and the assay tube, incubate in a 60°C water bath for 10 minutes. Immediately remove the tube and measure the absorbance of each tube using a 1 cm optical pathlength cuvette at 460 nm, zeroed with double-distilled water. The experimental results are shown in Figure 10. The MPO activity in the DSS model group was significantly higher than that in the normal control group (P<0.001). The MPO activity in the blank preparation group was similar to that in the model group, indicating that the excipients in the preparation had no anti-inflammatory effect. The MPO activity in mice with ulcerative colitis treated with HYP and HYP-M was significantly lower than that in the DSS model group (P<0.01), and the MPO activity in the HYP-M group was significantly lower than that in the HYP group (P<0.05). This indicates that HYP and HYP-M have the therapeutic effect of inhibiting colonic inflammation, and HYP-M can enhance the inhibitory effect of HYP on inflammation.

[0061] 5. Histopathological evaluation of mice

[0062] Eight days after inflammation, the mice were sacrificed, the abdominal cavity was rapidly opened, and the colon, kidney, and liver tissues were removed. The tissues were fixed with 10% neutral formalin for 24-48 hours, rinsed with running water overnight, dehydrated with graded alcohol solutions, cleared with xylene, dewaxed, embedded, and 5-micron sections were dewaxed and stained with hematoxylin-eosin (HE). HE staining steps: 1) Dewaxing and rehydration: After dewaxing in xylene for 5-10 minutes, switch to fresh xylene and continue dewaxing for 5-10 minutes. Then, place in 100%, 90%, and 70% ethanol for 2-5 minutes each, and then place in ultrapure sterile water for 2 minutes. 2) Staining: Stain with hematoxylin solution for 5-10 minutes, rinse with running water for 10-15 minutes, rinse with distilled water for several seconds, and then with 95% ethanol for 5 seconds. 3) Counterstaining: Stain with eosin solution for 1-2 minutes, rinse twice with water, and dehydrate twice with 95% ethanol. 4) Clearing: Clear with xylene for 5 minutes. 5) Sealing: Seal with neutral gum. The experimental results are shown in Figure 11, and Figure 11A is a cross-sectional image of the colon. The mucosal epithelial cell structure of the colon of mice in the normal control group was intact, without swelling, regular crypt structure, intact intestinal villus structure, and high goblet cell content. The colon of mice in the DSS group was eroded, the glandular structure was damaged, the crypt structure was irregular, the intestinal villi were deformed and damaged, goblet cells were missing, and there was a large amount of inflammatory cell infiltration. Compared with the model group, HF-HYP, LF-HYP, H-HYP-M, L-HPY-M and the positive control group all showed significant repair. At the same time, different doses of free HYP and HYP-M had different degrees of restoration of the tissue morphology of the mouse colon, and the high-dose group had better repair. Compared with the free HYP group, the HYP-M group had a better degree of restoration of the tissue morphology of the mouse colon, indicating that HYP-M can better improve the colon damage of mice with ulcerative colitis. Furthermore, no significant differences were observed in colon tissue between the blank and model control groups, further demonstrating that TPGS and F68 have no therapeutic effect on mice with colitis. As shown in Figure 11BC, while the blank control group showed normal cellular structure, the model group showed necrosis, cavitation, unclear cell outlines, condensed nuclei, and significant inflammatory cell infiltration, swelling, and degeneration, indicating that ulcerative colitis causes renal and liver damage. Compared with the model group, HF-HYP, LF-HYP, H-HYP-M, L-HPY-M, and the positive control group showed reduced necrotic cells and inflammatory cell infiltration, alleviating damage. Furthermore, different doses of free HYP and HYP-M showed varying degrees of protective effect on the histomorphology of the kidneys and livers in mice. More importantly, the histomorphology of mice in the HYP-M group was better than that in the free HYP group, indicating that HYP-M can better ameliorate renal and liver damage caused by ulcerative colitis. Furthermore, no significant differences were observed between the blank and model control groups, further demonstrating that TPGS and F68 have no therapeutic effect on the renal and liver damage caused by colitis in mice. In summary, HYP-M can effectively enhance the protective effect of HYP on the organs of mice with ulcerative colitis.

[0063] Based on the above results, it can be seen that the hyperoside polymer mixed micelles described in the present invention can improve the solubility and oral bioavailability of hyperoside, have a therapeutic effect on ulcerative colitis, and the therapeutic effect is better than that of hyperoside.

[0064] The above embodiment is used to produce sulfur. It only needs to be slightly modified on the basis of this embodiment, and a three-stage converter is added. Then it can be used to replace the conventional three-stage Claus process for producing sulfur.

[0065] The above is a further detailed description of the present invention and should not be considered as a limitation on the specific implementation of the present invention. For those skilled in the art, simple deductions or substitutions that do not depart from the concept of the present invention are within the scope of protection of the present invention.

Claims

1. A hyperoside polymer mixed micelle, characterized in that it comprises: Hyperoside, TPGS and F68, wherein the polymers TPGS and F68 are used as carriers of the mixed micelle, and hyperoside is encapsulated therein.

2. The hyperoside polymer mixed micelle according to claim 1, characterized in that: The mass ratio of the hyperoside, TPGS and F68 is 1:(2 - 6):

6.

3. The hyperoside polymer mixed micelle according to claim 1, characterized in that: The particle size of the polymer mixed micelle is 20 - 40 nm, the PDI value is 0.150 - 0.200, the potential value is -18 - -25 mV, the encapsulation efficiency is 85% - 95%, and the drug loading is 7% - 11%.

4. A preparation method of the hyperoside polymer mixed micelle according to any one of claims 1 to 3, characterized in that it comprises the following steps: (1) Dissolve hyperoside, TPGS and F68 in an organic solvent, and ultrasonically form a uniform mixed solution; (2) Control the temperature of the rotary evaporator to be 45 °C, and evaporate the organic solvent of the mixed solution to dryness to obtain a hyperoside film with TPGS and F68 as carriers; (3) Add the hyperoside film with TPGS and F68 as carriers to double-distilled water for hydration to obtain a hyperoside polymer mixed micelle preparation.

5. The preparation method according to claim 4, characterized in that: The organic solvent in step (1) is methanol.

6. An application of the hyperoside polymer mixed micelle according to any one of claims 1 to 3 in the preparation of a drug for treating ulcerative colitis.

7. The application according to claim 6, characterized in that: The daily dosage of the hyperoside polymer mixed micelle is 100 mg / kg - 50 mg / kg.

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