Microneedle patch for treating male erectile dysfunction and preparation and application thereof

Abstract

The invention discloses a microneedle patch for treating male erectile dysfunction as well as preparation and application of the microneedle patch. The microneedle patch comprises a soluble matrix carrier and a needle-shaped array, wherein the needle-shaped array is formed by ED treatment medicines and is wrapped in the soluble matrix carrier; the matrix carrier comprises a polymer or polysaccharide with high mechanical strength, high biocompatibility and high water solubility; the ED treatment medicine is prepared from tadalafil, sildenafil and vardenafil; the micro-needle patch is used for rapidly delivering a drug for treating male erectile dysfunction in a percutaneous manner, and the effect of treating male erectile dysfunction is achieved.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of biomedical material engineering, and more particularly relates to a microneedle patch for treating male erectile dysfunction and preparation and application thereof. BACKGROUND

[0002] PDE5i is a type 5 phosphodiesterase inhibitor, which is a class of drugs used to treat erectile dysfunction (ED), and its representative drugs include sildenafil citrate tablets, vardenafil hydrochloride tablets, tadalafil tablets, etc. Male erectile dysfunction is a relatively common sexual dysfunction, which refers to the intermittent or persistent inability to achieve erection. The pharmacological action of type 5 phosphodiesterase inhibitor is to inhibit the effect of PDE5 on smooth muscle relaxation, enhance the effect of cyclic guanosine monophosphate, and thus make the penis erect. Because the body releases nitric oxide during sexual stimulation, nitric oxide produces cyclic guanosine monophosphate, a chemical component that promotes penile erection, through a series of chemical changes, which can relax smooth muscle and allow blood to flow to the penis to make it erect. When PDE5 is produced in excess, it affects the effect of cyclic guanosine monophosphate on smooth muscle relaxation. Therefore, the inhibition of PDE5 by type 5 phosphodiesterase inhibitor can improve penile erectile dysfunction.

[0003] However, in current clinical applications, oral PDE5i drugs need to be administered in advance, and there are common systemic complications such as dizziness and blurred vision, and more than 50% of patients are not satisfied with the curative effect and have poor compliance. Therefore, although PDE5i is currently a recognized effective drug for ED patients, there is still an urgent need for a new type of transdermal drug delivery method that is efficient, convenient and minimally invasive. SUMMARY

[0004] (I) Technical problems solved

[0005] In view of the above background, the present application provides a microneedle patch for treating male erectile dysfunction, which is used for the delivery of drugs for treating male erectile dysfunction. The present application solves the problem that the existing erectile dysfunction treatment drugs need to be administered in advance, which is easy to cause systemic complications such as dizziness and blurred vision, and the drug effect is not obvious, thereby leading to poor compliance.

[0006] (II) Technical solutions

[0007] To achieve the above-mentioned purpose, the present application adopts the following technical solutions:

[0008] A microneedle patch for treating male erectile dysfunction, the microneedle patch comprising a needle-shaped array of a soluble matrix carrier and an ED treatment drug loaded therein; the matrix carrier comprises a polymer or polysaccharide with high mechanical strength, high biocompatibility and high water solubility; the microneedle patch is used for rapid transdermal delivery of drugs for treating male erectile dysfunction, and achieves the effect of treating male erectile dysfunction.

[0009] Further, the ED treatment drug is a type 5 phosphodiesterase inhibitor PDE5i, specifically including tadalafil, sildenafil, vardenafil, avanafil, ladalafil, mirodenafil, udenafil, etc.

[0010] The application also provides a preparation method of a microneedle patch for treating male erectile dysfunction, comprising the following steps:

[0011] Step 1: A certain amount of polymer or polysaccharide is weighed and dissolved in dimethyl sulfoxide, N, N-dimethylformamide, ethanol or the like, stirred until completely dissolved to form a matrix carrier solution, and an ED treatment drug is slowly added to the matrix carrier solution, and stirred at 500 rpm and 25°C for 2 hours to form a clear mixed dispersion;

[0012] Step 2: A polydimethylsiloxane PDMS mold with an inverted pyramid or conical structure micropore array is selected, and the mixed dispersion prepared in step 1 is added dropwise on the surface of the PDMS mold, and the mixed dispersion is ensured to completely cover and penetrate into all micropores;

[0013] Step 3: The mold is placed in a vacuum drying oven to remove bubbles and part of the moisture, and then transferred to a normal pressure drying oven for drying for 24-48 hours until the mixed dispersion is completely solidified to form a solid microneedle, and the mold is demolded to obtain a soluble microneedle patch loaded with an ED treatment drug.

[0014] Further, in step 1, the polymer is at least one of polyvinylpyrrolidone PVP, polyvinyl alcohol PVA and hyaluronic acid HA, and the polysaccharide is at least one of sulfobutyl ether-β-cyclodextrin and methyl cyclodextrin, or a mixture thereof, for example: PVP:HA=2:1 or 10:1.

[0015] Further, the mass ratio of the polymer or polysaccharide to the ED treatment drug ranges from 20:1 to 1:10.

[0016] Further, the microneedle height of the mold is 200-1000 μm, the bottom width is 100-300 μm, the needle tip angle is 10-30°, and the needle density is ~50-200 needle tips / cm²; preferably, the microneedle height of the mold is 800 μm, the bottom width is 300 μm, the needle tip angle is 30°, and the needle density is ~100 needle tips / cm².

[0017] Further, the vacuum degree in the vacuum drying oven is -0.01 MPa, and the temperature is 45°C; the temperature in the normal pressure drying oven is 45°C, and the relative humidity is <30%.

[0018] The application also provides application of the microneedle patch for treating male erectile dysfunction in preparation of a product for treating male erectile dysfunction.

[0019] The application also provides application of the ED treatment drug in preparation of the microneedle patch for treating male erectile dysfunction.

[0020] The application adopts a vacuum-assisted template method to prepare the soluble microneedle patch loaded with the ED treatment drug; when the patient with erectile dysfunction needs to take medicine, the polymer microneedle patch loaded with the PDE5i drug is placed in the inguinal region of the inner thigh, and the finger is gently pressed for about one minute, at this time, the needle tip is completely dissolved after the patch is removed, that is, the drug delivery is successful, the ED treatment drug is quickly and efficiently delivered, and thus the purpose of treating male erectile dysfunction is achieved.

[0021] (Three) beneficial effects

[0022] The beneficial effects of the application are:

[0023] The microneedle patch provided by the application is based on a soluble microneedle carrier, and compared with oral administration and cream, the soluble microneedle carrier can efficiently load PDE5i and has good strength, can effectively penetrate the skin and quickly release the drug. In terms of biological safety, the soluble microneedle also has excellent biocompatibility and little damage to the skin; in terms of drug loading and release capacity, the supramolecular soluble microneedle has good loading capacity for hydrophilic and hydrophobic drugs, and has high drug loading capacity, low local viscosity and fast diffusion rate in the skin. The application proves that the microneedle patch has the characteristics of high biocompatibility, low drug concentration and fast onset time, and adopts a new transdermal drug delivery method which is efficient, convenient and low-trauma, can effectively improve the compliance of patients with erectile dysfunction, improve the curative effect of patients and improve the quality of life of patients. BRIEF DESCRIPTION OF DRAWINGS

[0024] In order to more clearly illustrate the embodiments of the application or the technical solutions in the prior art, the following will briefly introduce the drawings needed to be used in the embodiments or the prior art description. Obviously, the drawings in the following description are only exemplary, and for those skilled in the art, other drawings can be obtained from the provided drawings without creative labor.

[0025] Please refer to the following blue format to supplement the schematic diagram generated during the preparation and effect experiment in the embodiments of the application

[0026] Figure 1 : The optical photograph of the microneedle patch for treating male erectile dysfunction prepared by the application;

[0027] Figure 2A scanning electron microscope photo of the micro-needle patch for treating male erectile dysfunction prepared by the present application;

[0028] Figure 3 A compression mechanical displacement curve of the micro-needle prepared by different proportions of polymer PVP and PDE5i drugs in the embodiments of the present application; in the figure, Load is force, and Distance is displacement;

[0029] Figure 4 An optical microscope photo of the micro-needle prepared by the polymer PVP and tadalafil with a mass ratio of 3:1 after being dissolved in gelatin hydrogel for different time;

[0030] Figure 5 Pictures of hemolysis of rats treated by different concentrations of drug-loaded micro-needles (PVP-TAD DMN);

[0031] Figure 6 Statistical results of hemolysis rate of rats treated by different concentrations of drug-loaded micro-needles PVP-TAD DMN; in the figure, hemolysis represents the hemolysis rate;

[0032] Figure 7 MOVAS cell viability treated by different concentrations of drug-loaded micro-needles PVP-TAD DMN; in the figure, MOVAS cell viability represents cell activity;

[0033] Figure 8 HUVEC cell viability treated by different concentrations of drug-loaded micro-needles PVP-TAD DMN; in the figure, HUVEC cell viability represents cell activity;

[0034] Figure 9 MOVAS cell live and dead staining treated by different concentrations of drug-loaded micro-needles PVP-TAD DMN;

[0035] Figure 10 HUVEC cell live and dead staining treated by different concentrations of drug-loaded micro-needles PVP-TAD DMN;

[0036] Figure 11 MOVAS cell survival rate statistics treated by different concentrations of drug-loaded micro-needles PVP-TAD DMN; in the figure, MOVAS calcein+ cells represent cell survival rate;

[0037] Figure 12 HUVEC cell survival rate statistics treated by different concentrations of drug-loaded micro-needles PVP-TAD DMN; in the figure, HUVEC calcein+ cells represent cell survival rate;

[0038] Figure 13For the treatment of 0 and 24h after the MOVAS cell scratch healing, wherein, Con represents the control group, HPA+AG represents the glycolipid group, PVP-TAD DMN represents the drug-loaded microneedle group, PVP DMN represents the empty microneedle group, and TAD represents the single drug group, i.e. oral drug group;

[0039] Figure 14 For the statistics of the MOVAS cell scratch healing rate after 24h treatment, Migration area represents the healing area in the figure;

[0040] Figure 15 For Ca2+ probe flow analysis of each group of MOVAS cells after treatment, Blank represents the blank group, Con represents the control group, HPA+AG represents the glycolipid group, PVP-TAD DMN represents the drug-loaded microneedle group, PVP DMN represents the empty microneedle group, and TAD represents the single drug group, i.e. oral drug group;

[0041] Figure 16 For Ca2+ probe flow analysis of each group of MOVAS cells after treatment, Blank represents the blank group, Con represents the control group, HPA+AG represents the glycolipid group, PVP-TAD DMN represents the drug-loaded microneedle group, PVP DMN represents the empty microneedle group, and TAD represents the single drug group, i.e. oral drug group; 2+ Fluorescence intensity statistics, MFI of Ca 2+ represents calcium ion content;

[0042] Figure 17 For each group of rats before and after administration of penile corpus cavernosum laser speckle blood flow imaging; Baseline represents before administration, and Post-administration represents after administration;

[0043] Figure 18 For each group of rats after administration of penile corpus cavernosum peak blood flow statistics, Perfusion represents blood flow;

[0044] Figure 19 For each group of rats after administration of penile corpus cavernosum time required to reach maximum blood flow statistics, Time represents the time required;

[0045] Figure 20 For each group of rats, the penile corpus cavernosum pressure diagram; wherein, Con represents the normal control group, DMED represents the erectile dysfunction DMED group, PVP DMN represents the DMED + empty microneedle group, PVP-TAD DMN represents the DMED + drug-loaded microneedle group, and TAD represents the DMED + oral drug group

[0046] Figure 21 For each group of rats, the penile corpus cavernosum pressure Max ICP / MAP statistics;

[0047] Figure 22 For each group of rats, the penile corpus cavernosum pressure Total ICP / MAP statistics

[0048] Figure 23 HE staining of major organs after 1 month of continuous application of PVP-TAD DMN microneedles, where Heart represents heart, Liver represents liver, Spleen represents spleen, Lung represents lung, Kidney represents kidney, Testis represents testis

[0049] It should be noted that in the above figures, English Blank in the figure represents the blank group, Con represents the control group, HPA+AG represents the glycolipid group, PVP-TAD DMN represents the drug-loaded microneedle group, PVP DMN represents the empty microneedle group, and TAD represents the single drug group, i.e. the oral drug group. DETAILED DESCRIPTION

[0050] The technical solutions in the embodiments of the present application will be described clearly and completely below in combination with the drawings in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor fall within the scope of protection of the present application.

[0051] Example 1: Preparation of microneedle patch

[0052] Step 1: weigh polyvinylpyrrolidone (PVP) 150 mg, dissolve in 0.5 mL DMSO, stir until completely dissolved. Weigh PDE5i 50 mg, slowly add to the PVP solution. Magnetic stirring (500 rpm, 25°C) for 2 hours to form a clear mixed solution. Step 2: select a polydimethylsiloxane (PDMS) mold with inverted pyramid or conical structure micropore array (microneedle height: 800 μm, bottom width: 300 μm, needle tip angle: 30°, needle density: ~100 needles / cm²). Step 3: Carefully drop the drug solution mixture prepared in step 1 on the surface of the PDMS mold, ensuring that the drug solution completely covers and penetrates all micropores. Place the mold in a vacuum drying oven (vacuum degree: -0.01 MPa, temperature: 45°C) to remove air bubbles and part of the water, about 15 minutes. Transfer to a normal pressure drying oven (temperature: 45°C, relative humidity: <30%) and dry for 24-48 hours until the drug solution is completely solidified to form solid microneedles. Demolding to obtain PDE5i-loaded soluble microneedle patch. The structure of the obtained microneedle patch is shown in Figure 1 and Figure 2 .

[0053] Characterization: The breaking force of microneedle array was determined using a universal testing machine. The single needle breaking force > 0.1 N, enough to penetrate the stratum corneum of human skin. In vitro dissolution and release: Microneedle dissolution experiment was performed using gelatin hydrogel. 3.0 g of gelatin was dissolved in 7 mL of water to prepare a gelatin hydrogel with a water content of 70%. Microneedles with a mass ratio of PVP and tadalafil of 3:1 were inserted into the gelatin hydrogel with a water content of 70%, and the microneedles were removed at different time points and photographed until the microneedles completely dissolved. Goal: Microneedles can completely dissolve within one minute. Results are shown in Figure 3 and Figure 4 .

[0054] Example Two: In vitro safety and functional verification of microneedle patch

[0055] 1. Cell culture

[0056] Mouse aortic vascular smooth muscle cells (MOVAS) were purchased from ATCC (American Type Culture Collection) and cultured in high-glucose Dulbecco's Modified Eagle Medium (DMEM, Boster) containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin (P / S). The cells were maintained in a growth state in a 37°C, 5% CO2 humidified incubator. Human umbilical vein endothelial cells (HUVEC) were purchased from Warner Bio and cultured in endothelial cell medium (ECM, ScienCell) supplemented with 5% fetal bovine serum (FBS), 1% endothelial cell growth supplement (ECGS), and 1% penicillin-streptomycin (P / S). The cells were also maintained in a 37°C, 5% CO2 humidified culture environment.

[0057] 2. Cytotoxicity evaluation

[0058] Microneedle extraction preparation: The tip part of the microneedle array was collected and placed in a graduated centrifuge tube; a small amount of pH 7.4 PBS buffer was added to make the theoretical PDE5i concentration greater than 5 mg / mL; ultrasonic treatment was performed at 37°C for 30 minutes (power 100 W, cycle 5 s on / 5 s off) to promote the swelling of the PVP matrix and the release of the drug from the needle tip; after ultrasonic treatment, centrifugation was performed at 3000 rpm for 10 min at 4°C, and the possible microneedle residue was discarded; the supernatant was filtered through a 0.22 µm sterile filter to obtain a clear needle tip extract. The extraction rate and concentration were verified by high performance liquid chromatography (HPLC), and the final concentration was adjusted to 5 mg / ml.

[0059] Hemolysis experiment: Healthy SD rats were selected, after intraperitoneal injection of sodium pentobarbital (30 mg / kg) anesthesia, under sterile conditions, cardiac puncture, collection of whole blood 5 mL into EDTA anticoagulant tube. Whole blood was centrifuged at 800 x g for 5 min, remove the upper plasma and white blood cell layer. Red blood cell precipitate was washed with sterile 0.9% saline 3 times (each time the same centrifugation conditions), finally prepared into 2% (v / v) red blood cell suspension. Respectively, 1 mL of red blood cell suspension was divided into sterile EP tube, a total of three groups were set up: negative control group (add normal saline), positive control group (add Triton X-100 to a final concentration of 0.1%), microneedle experimental group (add microneedle extract to a final concentration of 2-200 mg / L, calculated as PDE5i). Each tube was vortexed, incubated at 37°C constant temperature water bath for 1 h, immediately after incubation, centrifuged at 800 x g for 5 min. Respectively, 100 μL supernatant was transferred to 96-well plates, and the absorbance (OD value) was measured at 545 nm wavelength to quantify the released hemoglobin. The hemolysis rate was calculated as follows: hemolysis rate = (OD experimental group - OD negative control group) / (OD positive control group - OD negative control group) x 100%. The experimental results are shown in Figure 5 and Figure 6 .

[0060] Cell viability detection: MOVAS and HUVEC cells were inoculated in 96-well plates, respectively, and the complete culture medium was used in a humidified incubator (37°C, 5% CO2) to culture to about 70% of the adhesion rate. After discarding the original culture medium, the cells were divided into two groups for treatment: the control group was replaced with fresh complete culture medium, and the microneedle experimental group was replaced with complete culture medium with a final concentration of 2-200 mg / L microneedle extract (calculated as PDE5i). Each group had 6 replicate wells, and the incubation was continued for 24 h. Then, CCK-8 reagent (working concentration 10%) was directly added to each well. Incubate in the dark at 37°C for 2 h. The OD value was measured at 450 nm wavelength. The OD value of the cell-free culture medium well was used as a blank. Cell viability was calculated as follows: cell viability (%) = (OD experimental group - OD blank group) / (OD control group - OD blank group) x 100%. The experimental results are shown in Figure 7 and Figure 8 .

[0061] Live-dead staining: MOVAS and HUVEC cells were seeded in 6-well plates and cultured in their respective complete medium in a humidified incubator (37 °C, 5% CO2) until they reached approximately 70% confluency. After the original medium was removed, the cells were divided into two groups for treatment: the control group was replaced with fresh complete medium, and the microneedle experimental group was replaced with complete medium with a final concentration of 10-100 mg / L microneedle extract (calculated as PDE5i). After 24 h of culture, the medium was removed, and each group of cells was detected using a Calcein-AM / PI double staining kit (Beyotime). By counting the number of green (live) and red (dead) fluorescent cells, the cytotoxicity of the microneedle extract was evaluated, and the survival rate was calculated as green cell number / total cell number x 100%. The experimental results are shown in Figure 9 , Figure 10 , Figure 11 and Figure 12 .

[0062] 3. Cell function evaluation

[0063] Wound healing assay: MOVAS cells were seeded in 6-well plates and cultured in the corresponding complete medium in a humidified incubator (37 °C, 5% CO2) until they reached complete confluence. Then, a sterile 1 mL pipette tip was used to draw a straight and uniform scratch in the center of each well. After the medium was removed, each well was gently rinsed with preheated PBS to remove the detached cells and debris. After the PBS was removed, the cells were divided into five groups for treatment: the control group was added with serum-free DMEM high glucose medium, the glycolipid group was added with serum-free medium containing 400 µM palmitic acid (PA) and 30 mM glucose, the empty microneedle group was added with serum-free medium containing empty microneedle extract based on the glycolipid group, the drug-loaded microneedle group was added with serum-free medium containing 5 μM microneedle extract (calculated as PDE5i) based on the glycolipid group, and the single drug group was added with serum-free medium containing 5 μM PDE5i based on the glycolipid group. The baseline image of the scratch area was immediately taken using an inverted microscope. Subsequently, the cells were returned to the incubator, and the image of the same wound area was taken every 12 hours until the scratch in the control group was completely closed. The wound healing was quantitatively evaluated by measuring the degree of closure using ImageJ software. The experimental results are shown in Figure 13 and Figure 14 .

[0064] Ca2+ probe detection: Intracellular calcium signals in MOVAS cells were assessed using the calcium-sensitive fluorescent probe Fluo-4 AM (Beyotime). MOVAS cells were seeded in 6-well plates and cultured with the corresponding complete medium in a humidified incubator (37°C, 5% CO2) until the cell attachment rate reached approximately 70%. The cells were then divided into six treatment groups: blank and control groups were treated with complete medium; the glycolipid group was treated with complete medium containing 400µM PA and 30mM glucose; the empty microneedle group was treated with complete medium containing empty microneedle extract in addition to the glycolipid group; the drug-loaded microneedle group was treated with complete medium containing 5µM microneedle extract (denoted as PDE5i) in addition to the glycolipid group; and the single-drug group was treated with complete medium containing 5µM PDE5i in addition to the glycolipid group. After incubation, the medium was removed from each well and the cells were washed three times with PBS. Subsequently, 5µM Fluo-4 AM working solution was added and the cells were incubated at 37°C for 30 minutes to facilitate efficient probe loading. After the incubation, the cells were washed three times with PBS and incubated for another 20 minutes to ensure that Fluo-4 AM was completely hydrolyzed and converted into its fluorescently active form, Fluo-4. Fluorescence intensity was then immediately measured using flow cytometry. Calcium ion signals can be used as an indirect indicator of smooth muscle cell function. Figure 15 and Figure 16 .

[0065] In summary, the experimental results above show that (1) hemolysis test and CCK-8 cell viability test showed that 2–200 mg / L microneedle extract had no obvious toxicity to MOVAS and HUVEC cells; Calcein-AM / PI staining further confirmed that the cell survival rate was >99% under 10–100 mg / L conditions; (2) in the scratch test, the cell migration rate of the drug-loaded microneedle group was significantly improved compared with the empty microneedle group (p < 0.01), and there was no statistical difference between the two groups; (3) Fluo-4 AM was used to detect the calcium ion content of MOVAS cells, and the results were consistent with the scratch test, suggesting that the PDE5i released by the microneedles retained the original functional activity of smooth muscle cells.

[0066] Example 3: Animal Experiment and Efficacy Evaluation of Microneedle Patches

[0067] 1. Experimental Animals and Model Establishment

[0068] Adult male SD rats (body weight 250-300 g) were purchased from authorized suppliers. The rats were acclimated for one week under room temperature (23 ± 2℃), normal humidity (40%-60%) and 12h light / dark cycle conditions, and free access to food and water, and then transferred to the SPF level animal room. Streptozotocin (STZ) was dissolved in 0.1 mol / L citrate buffer to prepare type I diabetes model. After being transferred to the SPF environment, part of the rats were left as the control group, and the rest of the rats were injected with STZ at a dose of 50 mg / kg for 5 consecutive days; the control group was injected with the same volume of citrate buffer. One week after the last injection, the fasting blood glucose of the rats was detected, and if the blood glucose value was ≥16.7 mmol / L, it was determined that the type I diabetes model was successful. At different time points (4w, 8w) after the end of STZ injection, APO(-) rats were screened by the apomorphine (APO) experiment. The specific method was as follows: after subcutaneous injection of APO in the neck of the rats, the rats were placed in a dark environment for observation, and the occurrence of licking the penis, penis growth or penis head exposure (i.e. good erectile function, marked as APO(+)) was used as the evaluation index; rats without the above reactions were recorded as APO(-). Finally, APO(-) diabetic rats were defined as DMED rats for subsequent experiments.

[0069] 2. Animal grouping and drug administration scheme:

[0070] The rats were divided into five groups (n=8 / group): "normal control group (Con)", "DMED group", "DMED + empty microneedle group", "DMED + drug-loaded microneedle group", "DMED + oral drug group". The administration methods were as follows: "DMED + empty microneedle group" and "DMED + drug-loaded microneedle group" were respectively applied with corresponding microneedle array to the skin of the lower abdomen of the rats (sterilized after shaving) for 2 min and then removed; "DMED + oral drug group" was given a single gavage administration of the same dose of PDE5i.

[0071] 3. Efficacy monitoring

[0072] Laser speckle blood flow imaging: The RFLSI III blood perfusion imaging system (RWD Life Science) was used to evaluate the time to peak blood flow (Tmax) and peak blood flow (PBF) of the corpus cavernosum of the penis before and after administration of the "DMED + empty microneedle group", "DMED + drug-loaded microneedle group" and "DMED + oral drug group". The rats were maintained in a supine position under isoflurane anesthesia (induction concentration of 4-5%, maintenance concentration of 1.5-2%) and placed on a 37°C constant temperature heating pad. The glans penis was exposed by gently pressing the lower abdomen of the rat, and a clamp was applied at the base of the penis (at the male urogenital papilla) to fix the penis. After 2-5 minutes of stable organ positioning, the baseline blood flow under resting conditions was recorded. After administration, the changes in blood flow in the corpus cavernosum of the penis were continuously monitored and recorded for 60 minutes; the blood flow change value was the difference between the blood flow value at each time point after injection and the baseline blood flow value. At the same time, 100 μL of rat blood samples were collected by tail vein blood sampling at the set time points (10 min, 20 min, 30 min, 40 min, 60 min, 80 min, 100 min) after administration of the "DMED + drug-loaded microneedle group" and "DMED + oral drug group", and the blood drug concentrations at the corresponding time points were determined by HPLC. The experimental results are shown in Figure 17 , Figure 18 and Figure 19 .

[0073] Cavernous pressure measurement: The erectile function of the "normal control group (Con)", "DMED group", "DMED + empty microneedle group", "DMED + drug-loaded microneedle group" and "DMED + oral drug group" was evaluated by electrically stimulating the cavernous nerve (CN). The specific method was as follows: After the rats were anesthetized by intraperitoneal injection of sodium pentobarbital (30 mg / kg), the major pelvic ganglion (MPG) and the cavernous nerve (CN) were surgically exposed, and the corpus cavernosum was dissected. A 25-gauge heparinized needle (heparin concentration: 100 IU / mL) was inserted into the proximal corpus cavernosum to monitor the intracavernosal pressure (ICP); at the same time, a PE-50 catheter was inserted into the carotid artery and connected to a biological signal acquisition and processing system (BIOPAC MP160) to monitor the mean arterial pressure (MAP). Using a bipolar platinum electrode, electrical stimulation was applied to the connection between MPG and CN (parameters: frequency 15 Hz, voltage 5.0 V, pulse width 1 ms, duration 60 s). Erectile function was evaluated by the following indicators: 1) the ratio of intracavernosal pressure to mean arterial pressure (ICP / MAP) during electrical stimulation; 2) the area under the curve of intracavernosal pressure during electrical stimulation (i.e. total ICP). The experimental results are shown in Figure 20 , Figure 21 and Figure 22 .

[0074] The STZ-induced type I diabetic rat DMED model was constructed, and the empty microneedle, drug-loaded microneedle or the same dose of oral administration were given according to the grouping; Based on the above experimental results, it can be known that: (1) Laser speckle blood flow imaging shows that, compared with oral microneedles (~50 min), the drug-loaded microneedle group can significantly improve the corpus cavernosum blood flow within 20-30 min after administration (p<0.01), and the peak blood flow is equivalent to that of the oral administration group; (2) In the cavernous pressure test, the erectile indicators (Max ICP / MAP, Total ICP / MAP) of the drug-loaded microneedle group also have no significant difference with the oral group; (3) Pharmacokinetic analysis shows that the time of PDE5i reaching the peak plasma concentration (Tmax) by microneedle administration is significantly shorter than that by oral administration, showing faster onset characteristics.

[0075] 4. Biological safety evaluation

[0076] Healthy SD rats were selected, and the same part of the lower abdomen was treated with a single microneedle patch every day for 30 days. The animals were sacrificed after the treatment. The main organs such as heart, liver, spleen, lung, kidney and testis were completely removed, fixed in 4% paraformaldehyde (Servicebio) at 4°C for 24 hours, then paraffin-embedded and prepared into 5 μm thick continuous sections. Hematoxylin-eosin (H&E) staining (Servicebio) was performed according to the standard operation procedure. At the same time, the full-thickness skin tissue in the microneedle patch area was collected, fixed, embedded and sectioned, and then the inflammatory factors (IL-1β, IL-6, TNF-α and TGF-β) were detected according to the standard immunohistochemical protocol. The primary antibody was incubated at 4°C overnight (Servicebio). After all the sections were sealed with neutral gum, histological evaluation was performed under an optical microscope, focusing on observing the abnormal structure of organ tissue, the degree of inflammatory cell infiltration and cell degeneration / necrosis and other toxicity indicators.

[0077] Biological safety: After the rats were treated with drug-loaded microneedle patches for 30 consecutive days, H&E staining of the main organs (heart, liver, spleen, lung, kidney and testis) and immunohistochemistry (IL-1β, IL-6, TNF-α and TGF-β) of the skin tissue showed no pathological changes and inflammatory cell infiltration, proving that the drug delivery system has good biocompatibility and safety. Figure 23 .

[0078] In summary, the present application provides a microneedle patch for treating male erectile dysfunction, which is used for the delivery of drugs for treating male erectile dysfunction. The problems of the existing erectile dysfunction treatment drugs, such as the need for planned advance administration, easy to cause dizziness, blurred vision and other systemic complications, poor drug efficacy and poor compliance, are solved.

[0079] Finally, it should be noted that the above examples are only used to illustrate the present application and do not limit the protection scope of the present application. In addition, after reading the technical content of the present application, those skilled in the art can make various modifications, modifications or variations to the present application, and all these equivalent forms also belong to the protection scope defined by the present application.

Claims

1. A microneedle patch for treating male erectile dysfunction, characterized in that: The microneedle patch includes a needle array consisting of a soluble matrix carrier and an ED treatment drug encapsulated therein; the matrix carrier includes a polymer or polysaccharide with high mechanical strength, high biocompatibility, and high water solubility; the microneedle patch is used for rapid transdermal delivery of drugs for treating male erectile dysfunction, thereby achieving the effect of treating male erectile dysfunction.

2. The microneedle patch for treating male erectile dysfunction according to claim 1, wherein: The ED treatment drugs are phosphodiesterase type 5 inhibitors PDE5i drugs, including tadalafil, sildenafil, vardenafil, avanafil, lodenafil, mironafil, and udenafil.

3. The method for preparing a microneedle patch for treating male erectile dysfunction according to claim 1, characterized in that: The following steps are involved: Step 1: Weigh a certain amount of polymer or polysaccharide, dissolve it in dimethyl sulfoxide, N,N-dimethylformamide or ethanol solvent, and stir until completely dissolved to form a matrix carrier solution; weigh the ED treatment drug, add it to the matrix carrier solution, and dissolve it to form a uniform mixed dispersion; Step 2: Select a polydimethylsiloxane (PDMS) mold with an inverted pyramid or conical micropore array, and pour the mixed dispersion prepared in step 1 onto the mold surface to ensure that the mixed dispersion completely covers and penetrates all micropores; Step 3: Place the mold in a vacuum drying oven to preliminarily remove the solvent, transfer it to a normal pressure drying oven and dry it for 24-48 hours until the mixed dispersion is completely dried and solidified to form solid microneedles, and demold to obtain a soluble microneedle patch loaded with ED treatment drugs.

4. The method for preparing a microneedle patch for treating male erectile dysfunction according to claim 3, characterized in that: In step 1, the polymer is at least one of polyvinyl pyrrolidone (PVP), polyvinyl alcohol (PVA), and hyaluronic acid (HA), and the polysaccharide is at least one of sulfobutyl ether-β-cyclodextrin and methyl cyclodextrin, or a mixture thereof.

5. The method for preparing a microneedle patch for treating male erectile dysfunction according to claim 3, characterized in that: The ED treatment drugs are phosphodiesterase type 5 inhibitors PDE5i drugs, including tadalafil, sildenafil, vardenafil, avanafil, lodenafil, mironafil, and udenafil.

6. The method for preparing a microneedle patch for treating male erectile dysfunction according to claim 3, characterized in that: The mass ratio of the polymer or polysaccharide to the ED therapeutic drug is in the range of 20:1 to 1:

10.

7. The method for preparing a microneedle patch for treating male erectile dysfunction according to claim 3, characterized in that: The microneedle height of the mold is 200-1000 μm, the bottom width is 100-300 μm, the needle tip angle is 10-30°, and the needle density is ~50-200 needle tips / cm².

8. Use of the microneedle patch for treating male erectile dysfunction according to any one of claims 1 to 3 in the preparation of a product for treating male erectile dysfunction.

9. Use of the ED therapeutic drug according to claim 2 in the preparation of a microneedle patch for treating male erectile dysfunction.

Images